Identification	NN	O	O
of	NN	O	O
APC2	NN	O	O
,	NN	O	O
a	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
tumour	NN	O	I-Disease
suppressor	NN	O	O
.	NN	O	O

The	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	I-Disease
APC	NN	O	I-Disease
)	NN	O	I-Disease
tumour	NN	O	I-Disease
-	NN	O	O
suppressor	NN	O	O
protein	NN	O	O
controls	NN	O	O
the	NN	O	O
Wnt	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
by	NN	O	O
forming	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
glycogen	NN	O	O
synthase	NN	O	O
kinase	NN	O	O
3beta	NN	O	O
(	NN	O	O
GSK	NN	O	O
-	NN	O	O
3beta	NN	O	O
)	NN	O	O
,	NN	O	O
axin	NN	O	O
/	NN	O	O
conductin	NN	O	O
and	NN	O	O
betacatenin	NN	O	O
.	NN	O	O

Complex	NN	O	O
formation	NN	O	O
induces	NN	O	O
the	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
of	NN	O	O
betacatenin	NN	O	O
.	NN	O	O

In	NN	O	O
colon	NN	O	B-Disease
carcinoma	NN	O	I-Disease
cells	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
APC	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
betacatenin	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
Tcf	NN	O	O
-	NN	O	O
4	NN	O	O
transcription	NN	O	O
factor	NN	O	O
(	NN	O	O
reviewed	NN	O	O
in	NN	O	O
[	NN	O	O
1	NN	O	O
]	NN	O	O
[	NN	O	O
2	NN	O	O
]	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
and	NN	O	O
genomic	NN	O	O
structure	NN	O	O
of	NN	O	O
APC	NN	O	O
homologues	NN	O	O
.	NN	O	O

Mammalian	NN	O	O
APC2	NN	O	O
,	NN	O	O
which	NN	O	O
closely	NN	O	O
resembles	NN	O	O
APC	NN	O	O
in	NN	O	O
overall	NN	O	O
domain	NN	O	O
structure	NN	O	O
,	NN	O	O
was	NN	O	O
functionally	NN	O	O
analyzed	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
two	NN	O	O
SAMP	NN	O	O
domains	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
conductin	NN	O	O
.	NN	O	O

Like	NN	O	O
APC	NN	O	O
,	NN	O	O
APC2	NN	O	O
regulates	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
active	NN	O	O
betacatenin	NN	O	O
-	NN	O	O
Tcf	NN	O	O
complexes	NN	O	O
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
using	NN	O	O
transient	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
assays	NN	O	O
in	NN	O	O
APC	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
colon	NN	O	B-Disease
carcinoma	NN	O	I-Disease
cells	NN	O	O
.	NN	O	O

Human	NN	O	O
APC2	NN	O	O
maps	NN	O	O
to	NN	O	O
chromosome	NN	O	O
19p13	NN	O	O
.	NN	O	O

3	NN	O	O
.	NN	O	O

APC	NN	O	O
and	NN	O	O
APC2	NN	O	O
may	NN	O	O
therefore	NN	O	O
have	NN	O	O
comparable	NN	O	O
functions	NN	O	O
in	NN	O	O
development	NN	O	O
and	NN	O	O
cancer	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
common	NN	O	O
MSH2	NN	O	O
mutation	NN	O	O
in	NN	O	O
English	NN	O	O
and	NN	O	O
North	NN	O	O
American	NN	O	O
HNPCC	NN	O	B-Disease
families	NN	O	O
:	NN	O	O
origin	NN	O	O
,	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
sex	NN	O	O
specific	NN	O	O
differences	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
,	NN	O	O
origin	NN	O	O
,	NN	O	O
and	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
germline	NN	O	O
MSH2	NN	O	O
gene	NN	O	O
mutation	NN	O	O
previously	NN	O	O
identified	NN	O	O
in	NN	O	O
seven	NN	O	O
kindreds	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
non	NN	O	I-Disease
-	NN	O	I-Disease
polyposis	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
HNPCC	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
(	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
at	NN	O	O
nt943	NN	O	O
+	NN	O	O
3	NN	O	O
)	NN	O	O
disrupts	NN	O	O
the	NN	O	O
3	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
exon	NN	O	O
5	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
this	NN	O	O
exon	NN	O	O
from	NN	O	O
MSH2	NN	O	O
mRNA	NN	O	O
and	NN	O	O
represents	NN	O	O
the	NN	O	O
only	NN	O	O
frequent	NN	O	O
MSH2	NN	O	O
mutation	NN	O	O
so	NN	O	O
far	NN	O	O
reported	NN	O	O
.	NN	O	O

Although	NN	O	O
this	NN	O	O
mutation	NN	O	O
was	NN	O	O
initially	NN	O	O
detected	NN	O	O
in	NN	O	O
four	NN	O	O
of	NN	O	O
33	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
analysed	NN	O	O
from	NN	O	O
eastern	NN	O	O
England	NN	O	O
,	NN	O	O
more	NN	O	O
extensive	NN	O	O
analysis	NN	O	O
has	NN	O	O
reduced	NN	O	O
the	NN	O	O
frequency	NN	O	O
to	NN	O	O
four	NN	O	O
of	NN	O	O
52	NN	O	O
(	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
English	NN	O	O
HNPCC	NN	O	B-Disease
kindreds	NN	O	O
analysed	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
MSH2	NN	O	O
mutation	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
10	NN	O	O
of	NN	O	O
20	NN	O	O
(	NN	O	O
50	NN	O	O
%	NN	O	O
)	NN	O	O
separately	NN	O	O
identified	NN	O	O
colorectal	NN	O	B-Disease
families	NN	O	O
from	NN	O	O
Newfoundland	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
from	NN	O	O
England	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
Newfoundland	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
10	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
haplotype	NN	O	O
analysis	NN	O	O
using	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
linked	NN	O	O
to	NN	O	O
MSH2	NN	O	O
was	NN	O	O
performed	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
English	NN	O	O
and	NN	O	O
US	NN	O	O
families	NN	O	O
there	NN	O	O
was	NN	O	O
little	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
recent	NN	O	O
common	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
MSH2	NN	O	O
splice	NN	O	O
site	NN	O	O
mutation	NN	O	O
in	NN	O	O
most	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
common	NN	O	O
haplotype	NN	O	O
was	NN	O	O
identified	NN	O	O
at	NN	O	O
the	NN	O	O
two	NN	O	O
flanking	NN	O	O
markers	NN	O	O
(	NN	O	O
CA5	NN	O	O
and	NN	O	O
D2S288	NN	O	O
)	NN	O	O
in	NN	O	O
eight	NN	O	O
of	NN	O	O
the	NN	O	O
Newfoundland	NN	O	O
families	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggested	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
within	NN	O	O
Newfoundland	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
reported	NN	O	O
by	NN	O	O
others	NN	O	O
for	NN	O	O
two	NN	O	O
MLH1	NN	O	O
mutations	NN	O	O
in	NN	O	O
Finnish	NN	O	O
HNPCC	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
calculated	NN	O	O
age	NN	O	O
related	NN	O	O
risks	NN	O	O
of	NN	O	O
all	NN	O	O
,	NN	O	O
colorectal	NN	O	B-Disease
,	NN	O	I-Disease
endometrial	NN	O	I-Disease
,	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
in	NN	O	O
nt943	NN	O	O
+	NN	O	O
3	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
MSH2	NN	O	O
mutation	NN	O	O
carriers	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
76	NN	O	O
)	NN	O	O
for	NN	O	O
all	NN	O	O
patients	NN	O	O
and	NN	O	O
for	NN	O	O
men	NN	O	O
and	NN	O	O
women	NN	O	O
separately	NN	O	O
.	NN	O	O

For	NN	O	O
both	NN	O	O
sexes	NN	O	O
combined	NN	O	O
,	NN	O	O
the	NN	O	O
penetrances	NN	O	O
at	NN	O	O
age	NN	O	O
60	NN	O	O
years	NN	O	O
for	NN	O	O
all	NN	O	O
cancers	NN	O	B-Disease
and	NN	O	O
for	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
were	NN	O	O
0	NN	O	O
.	NN	O	O

86	NN	O	O
and	NN	O	O
0	NN	O	O
.	NN	O	O

57	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
risk	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
01	NN	O	O
)	NN	O	O
in	NN	O	O
males	NN	O	O
than	NN	O	O
females	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
63	NN	O	O
v	NN	O	O
0	NN	O	O
.	NN	O	O
30	NN	O	O
and	NN	O	O
0	NN	O	O
.	NN	O	O
84	NN	O	O
v	NN	O	O
0	NN	O	O
.	NN	O	O
44	NN	O	O
at	NN	O	O
ages	NN	O	O
50	NN	O	O
and	NN	O	O
60	NN	O	O
years	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

For	NN	O	O
females	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
high	NN	O	O
risk	NN	O	O
of	NN	O	O
endometrial	NN	O	B-Disease
cancer	NN	O	I-Disease
(	NN	O	O
0	NN	O	O
.	NN	O	O
5	NN	O	O
at	NN	O	O
age	NN	O	O
60	NN	O	O
years	NN	O	O
)	NN	O	O
and	NN	O	O
premenopausal	NN	O	B-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
(	NN	O	O
0	NN	O	O
.	NN	O	O
2	NN	O	O
at	NN	O	O
50	NN	O	O
years	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
intersex	NN	O	O
differences	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
risks	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
screening	NN	O	O
programmes	NN	O	O
and	NN	O	O
for	NN	O	O
attempts	NN	O	O
to	NN	O	O
identify	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
modifiers	NN	O	O
.	NN	O	O

Age	NN	O	O
of	NN	O	O
onset	NN	O	O
in	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
sex	NN	O	O
specific	NN	O	O
influence	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
E	NN	O	O
genotype	NN	O	O
and	NN	O	O
normal	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
length	NN	O	O
.	NN	O	O

Age	NN	O	O
of	NN	O	O
onset	NN	O	O
(	NN	O	O
AO	NN	O	O
)	NN	O	O
of	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
length	NN	O	O
of	NN	O	O
an	NN	O	O
expanded	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Apolipoprotein	NN	O	O
E	NN	O	O
(	NN	O	O
APOE	NN	O	O
)	NN	O	O
genotype	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
influence	NN	O	O
AO	NN	O	O
in	NN	O	O
Alzheimer	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
rendering	NN	O	O
the	NN	O	O
APOE	NN	O	O
gene	NN	O	O
a	NN	O	O
likely	NN	O	O
candidate	NN	O	O
to	NN	O	O
affect	NN	O	O
AO	NN	O	O
in	NN	O	O
other	NN	O	O
neurological	NN	O	B-Disease
diseases	NN	O	I-Disease
too	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
determined	NN	O	O
APOE	NN	O	O
genotype	NN	O	O
and	NN	O	O
normal	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
length	NN	O	O
in	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
for	NN	O	O
138	NN	O	O
HD	NN	O	B-Disease
patients	NN	O	O
who	NN	O	O
were	NN	O	O
previously	NN	O	O
analysed	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
length	NN	O	O
.	NN	O	O

Genotyping	NN	O	O
for	NN	O	O
APOE	NN	O	O
was	NN	O	O
performed	NN	O	O
blind	NN	O	O
to	NN	O	O
clinical	NN	O	O
information	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
highlighting	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
repeat	NN	O	O
length	NN	O	O
upon	NN	O	O
AO	NN	O	O
in	NN	O	O
maternally	NN	O	O
inherited	NN	O	O
HD	NN	O	B-Disease
and	NN	O	O
in	NN	O	O
male	NN	O	O
patients	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
APOE	NN	O	O
epsilon2epsilon3	NN	O	O
genotype	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
significantly	NN	O	O
earlier	NN	O	O
AO	NN	O	O
in	NN	O	O
males	NN	O	O
than	NN	O	O
in	NN	O	O
females	NN	O	O
.	NN	O	O

Such	NN	O	O
a	NN	O	O
sex	NN	O	O
difference	NN	O	O
in	NN	O	O
AO	NN	O	O
was	NN	O	O
not	NN	O	O
apparent	NN	O	O
for	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
APOE	NN	O	O
genotypes	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
subtle	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
neurodegeneration	NN	O	B-Disease
in	NN	O	O
HD	NN	O	B-Disease
may	NN	O	O
allow	NN	O	O
interacting	NN	O	O
genes	NN	O	O
to	NN	O	O
exert	NN	O	O
gender	NN	O	O
specific	NN	O	O
effects	NN	O	O
upon	NN	O	O
AO	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
recurrent	NN	O	O
bacteremic	NN	O	B-Disease
infections	NN	O	I-Disease
due	NN	O	I-Disease
to	NN	O	I-Disease
Neisseria	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
serum	NN	O	O
of	NN	O	O
a	NN	O	O
29	NN	O	O
-	NN	O	O
year	NN	O	O
old	NN	O	O
woman	NN	O	O
with	NN	O	O
a	NN	O	O
recent	NN	O	O
episode	NN	O	O
of	NN	O	O
disseminated	NN	O	B-Disease
gonococcal	NN	O	I-Disease
infection	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
and	NN	O	O
arthritis	NN	O	B-Disease
as	NN	O	O
a	NN	O	O
child	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
lack	NN	O	O
serum	NN	O	O
hemolytic	NN	O	O
complement	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
seventh	NN	O	O
component	NN	O	O
of	NN	O	O
complement	NN	O	O
(	NN	O	O
C7	NN	O	O
)	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
by	NN	O	O
functional	NN	O	O
or	NN	O	O
immunochemical	NN	O	O
assays	NN	O	O
,	NN	O	O
whereas	NN	O	O
other	NN	O	O
components	NN	O	O
were	NN	O	O
normal	NN	O	O
by	NN	O	O
hemolytic	NN	O	O
and	NN	O	O
immunochemical	NN	O	O
assessment	NN	O	O
.	NN	O	O

Her	NN	O	O
fresh	NN	O	O
serum	NN	O	O
lacked	NN	O	O
complement	NN	O	O
-	NN	O	O
mediated	NN	O	O
bactericidal	NN	O	O
activity	NN	O	O
against	NN	O	O
Neisseria	NN	O	O
gonorrhoeae	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
fresh	NN	O	O
normal	NN	O	O
serum	NN	O	O
or	NN	O	O
purified	NN	O	O
C7	NN	O	O
restored	NN	O	O
bactericidal	NN	O	O
activity	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
hemolytic	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
functional	NN	O	I-Disease
C7	NN	O	I-Disease
activity	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Opsonization	NN	O	O
and	NN	O	O
generation	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
functioned	NN	O	O
normally	NN	O	O
.	NN	O	O

Complete	NN	O	B-Disease
absence	NN	O	I-Disease
of	NN	O	I-Disease
C7	NN	O	I-Disease
was	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
sibling	NN	O	O
who	NN	O	O
had	NN	O	O
the	NN	O	O
clinical	NN	O	O
syndrome	NN	O	O
of	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
and	NN	O	O
arthritis	NN	O	B-Disease
as	NN	O	O
a	NN	O	O
child	NN	O	O
and	NN	O	O
in	NN	O	O
this	NN	O	O
siblings	NN	O	O
clinically	NN	O	O
well	NN	O	O
eight	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
son	NN	O	O
.	NN	O	O

HLA	NN	O	O
histocompatibility	NN	O	O
typing	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
members	NN	O	O
did	NN	O	O
not	NN	O	O
demonstrate	NN	O	O
evidence	NN	O	O
for	NN	O	O
genetic	NN	O	O
linkage	NN	O	O
of	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
loci	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
cases	NN	O	O
of	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
infectious	NN	O	O
complications	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
bactericidal	NN	O	O
activity	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
host	NN	O	O
defense	NN	O	O
against	NN	O	O
bacteremic	NN	O	B-Disease
neisseria	NN	O	I-Disease
infections	NN	O	I-Disease
.	NN	O	O

Increased	NN	O	O
incidence	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
in	NN	O	O
patients	NN	O	O
with	NN	O	O
cartilage	NN	O	B-Disease
-	NN	O	I-Disease
hair	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
.	NN	O	O

OBJECTIVE	NN	O	O
Previous	NN	O	O
reports	NN	O	O
have	NN	O	O
suggested	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
among	NN	O	O
patients	NN	O	O
with	NN	O	O
cartilage	NN	O	B-Disease
-	NN	O	I-Disease
hair	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
(	NN	O	O
CHH	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
to	NN	O	O
further	NN	O	O
evaluate	NN	O	O
this	NN	O	O
risk	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
CHH	NN	O	B-Disease
and	NN	O	O
their	NN	O	O
first	NN	O	O
-	NN	O	O
degree	NN	O	O
relatives	NN	O	O
.	NN	O	O

STUDY	NN	O	O
DESIGN	NN	O	O
One	NN	O	O
hundred	NN	O	O
twenty	NN	O	O
-	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
CHH	NN	O	B-Disease
were	NN	O	O
identified	NN	O	O
through	NN	O	O
2	NN	O	O
countrywide	NN	O	O
epidemiologic	NN	O	O
surveys	NN	O	O
in	NN	O	O
1974	NN	O	O
and	NN	O	O
in	NN	O	O
1986	NN	O	O
.	NN	O	O

Their	NN	O	O
parents	NN	O	O
and	NN	O	O
nonaffected	NN	O	O
siblings	NN	O	O
were	NN	O	O
identified	NN	O	O
through	NN	O	O
the	NN	O	O
Population	NN	O	O
Register	NN	O	O
Center	NN	O	O
.	NN	O	O

This	NN	O	O
cohort	NN	O	O
underwent	NN	O	O
follow	NN	O	O
-	NN	O	O
up	NN	O	O
for	NN	O	O
cancer	NN	O	B-Disease
incidence	NN	O	O
through	NN	O	O
the	NN	O	O
Finnish	NN	O	O
Cancer	NN	O	O
Registry	NN	O	O
to	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
1995	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
A	NN	O	O
statistically	NN	O	O
significant	NN	O	O
excess	NN	O	O
risk	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
was	NN	O	O
seen	NN	O	O
among	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
CHH	NN	O	B-Disease
(	NN	O	O
standardized	NN	O	O
incidence	NN	O	O
ratio	NN	O	O
6	NN	O	O
.	NN	O	O
9	NN	O	O
,	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
2	NN	O	O
.	NN	O	O
3	NN	O	O
to	NN	O	O
16	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
mainly	NN	O	O
attributable	NN	O	O
to	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
Hodgkins	NN	O	I-Disease
lymphoma	NN	O	I-Disease
(	NN	O	O
standardized	NN	O	O
incidence	NN	O	O
ratio	NN	O	O
90	NN	O	O
,	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
18	NN	O	O
to	NN	O	O
264	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
excess	NN	O	O
risk	NN	O	O
of	NN	O	O
basal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
was	NN	O	O
seen	NN	O	O
(	NN	O	O
standardized	NN	O	O
incidence	NN	O	O
ratio	NN	O	O
35	NN	O	O
,	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
7	NN	O	O
.	NN	O	O
2	NN	O	O
to	NN	O	O
102	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
cancer	NN	O	B-Disease
incidence	NN	O	O
among	NN	O	O
the	NN	O	O
siblings	NN	O	O
or	NN	O	O
the	NN	O	O
parents	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
from	NN	O	O
the	NN	O	O
average	NN	O	O
cancer	NN	O	B-Disease
incidence	NN	O	O
in	NN	O	O
the	NN	O	O
Finnish	NN	O	O
population	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
This	NN	O	O
study	NN	O	O
confirms	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
especially	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
Hodgkins	NN	O	I-Disease
lymphoma	NN	O	I-Disease
,	NN	O	O
probably	NN	O	O
attributable	NN	O	O
to	NN	O	O
defective	NN	O	O
immunity	NN	O	O
,	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
CHH	NN	O	B-Disease
.	NN	O	O

Genotype	NN	O	O
and	NN	O	O
phenotype	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
dihydropyrimidine	NN	O	B-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Dihydropyrimidine	NN	O	B-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
DPD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
characterised	NN	O	O
by	NN	O	O
thymine	NN	O	O
-	NN	O	O
uraciluria	NN	O	O
in	NN	O	O
homozygous	NN	O	O
deficient	NN	O	O
patients	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
variable	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
genetic	NN	O	O
and	NN	O	O
phenotypic	NN	O	O
basis	NN	O	O
for	NN	O	O
DPD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
have	NN	O	O
reviewed	NN	O	O
17	NN	O	O
families	NN	O	O
presenting	NN	O	O
22	NN	O	O
patients	NN	O	O
with	NN	O	O
complete	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
DPD	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
this	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
,	NN	O	O
7	NN	O	O
different	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
including	NN	O	O
2	NN	O	O
deletions	NN	O	O
[	NN	O	O
295	NN	O	O
-	NN	O	O
298delTCAT	NN	O	O
,	NN	O	O
1897delC	NN	O	O
]	NN	O	O
,	NN	O	O
1	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
[	NN	O	O
IVS14	NN	O	O
+	NN	O	O
1G	NN	O	O
>	NN	O	O
A	NN	O	O
)	NN	O	O
]	NN	O	O
and	NN	O	O
4	NN	O	O
missense	NN	O	O
mutations	NN	O	O
(	NN	O	O
85T	NN	O	O
>	NN	O	O
C	NN	O	O
,	NN	O	O
703C	NN	O	O
>	NN	O	O
T	NN	O	O
,	NN	O	O
2658G	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
2983G	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
the	NN	O	O
various	NN	O	O
mutations	NN	O	O
among	NN	O	O
DPD	NN	O	B-Disease
patients	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
invariant	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
is	NN	O	O
by	NN	O	O
far	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
(	NN	O	O
52	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
other	NN	O	O
six	NN	O	O
mutations	NN	O	O
are	NN	O	O
less	NN	O	O
frequently	NN	O	O
observed	NN	O	O
.	NN	O	O

A	NN	O	O
large	NN	O	O
phenotypic	NN	O	O
variability	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
,	NN	O	O
with	NN	O	O
convulsive	NN	O	B-Disease
disorders	NN	O	I-Disease
,	NN	O	O
motor	NN	O	B-Disease
retardation	NN	O	I-Disease
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
being	NN	O	O
the	NN	O	O
most	NN	O	O
abundant	NN	O	O
manifestations	NN	O	O
.	NN	O	O

A	NN	O	O
clear	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
genotype	NN	O	O
and	NN	O	O
phenotype	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
.	NN	O	O

An	NN	O	O
altered	NN	O	O
beta	NN	O	O
-	NN	O	O
alanine	NN	O	O
,	NN	O	O
uracil	NN	O	O
and	NN	O	O
thymine	NN	O	O
homeostasis	NN	O	O
might	NN	O	O
underlie	NN	O	O
the	NN	O	O
various	NN	O	O
clinical	NN	O	B-Disease
abnormalities	NN	O	I-Disease
encountered	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
DPD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Fibroblast	NN	O	O
growth	NN	O	O
factor	NN	O	O
homologous	NN	O	O
factor	NN	O	O
2	NN	O	O
(	NN	O	O
FHF2	NN	O	O
)	NN	O	O
:	NN	O	O
gene	NN	O	O
structure	NN	O	O
,	NN	O	O
expression	NN	O	O
and	NN	O	O
mapping	NN	O	O
to	NN	O	O
the	NN	O	O
Borjeson	NN	O	B-Disease
-	NN	O	I-Disease
Forssman	NN	O	I-Disease
-	NN	O	I-Disease
Lehmann	NN	O	I-Disease
syndrome	NN	O	I-Disease
region	NN	O	O
in	NN	O	O
Xq26	NN	O	O
delineated	NN	O	O
by	NN	O	O
a	NN	O	O
duplication	NN	O	O
breakpoint	NN	O	O
in	NN	O	O
a	NN	O	O
BFLS	NN	O	B-Disease
-	NN	O	O
like	NN	O	O
patient	NN	O	O
.	NN	O	O

Borjeson	NN	O	B-Disease
-	NN	O	I-Disease
Forssman	NN	O	I-Disease
-	NN	O	I-Disease
Lehmann	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
BFLS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
syndromal	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
mental	NN	O	I-Disease
retardation	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
maps	NN	O	O
by	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
q26	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
male	NN	O	O
patient	NN	O	O
with	NN	O	O
BFLS	NN	O	B-Disease
-	NN	O	O
like	NN	O	O
features	NN	O	O
and	NN	O	O
a	NN	O	O
duplication	NN	O	O
,	NN	O	O
46	NN	O	O
,	NN	O	O
Y	NN	O	O
,	NN	O	O
dup	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
(	NN	O	O
q26q28	NN	O	O
)	NN	O	O
,	NN	O	O
inherited	NN	O	O
from	NN	O	O
his	NN	O	O
phenotypically	NN	O	O
normal	NN	O	O
mother	NN	O	O
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridisation	NN	O	O
using	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosome	NN	O	O
clones	NN	O	O
from	NN	O	O
Xq26	NN	O	O
localised	NN	O	O
the	NN	O	O
duplication	NN	O	O
breakpoint	NN	O	O
to	NN	O	O
an	NN	O	O
approximately	NN	O	O
400	NN	O	O
-	NN	O	O
kb	NN	O	O
interval	NN	O	O
in	NN	O	O
the	NN	O	O
Xq26	NN	O	O
.	NN	O	O

3	NN	O	O
region	NN	O	O
between	NN	O	O
DXS155	NN	O	O
and	NN	O	O
DXS294	NN	O	O
/	NN	O	O
DXS730	NN	O	O
.	NN	O	O

Database	NN	O	O
searches	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
available	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
from	NN	O	O
the	NN	O	O
region	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
fibroblast	NN	O	O
growth	NN	O	O
factor	NN	O	O
homologous	NN	O	O
factor	NN	O	O
gene	NN	O	O
,	NN	O	O
FHF2	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
duplication	NN	O	O
breakpoint	NN	O	O
interval	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
structure	NN	O	O
of	NN	O	O
FHF2	NN	O	O
was	NN	O	O
determined	NN	O	O
and	NN	O	O
two	NN	O	O
new	NN	O	O
exons	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
new	NN	O	O
5	NN	O	O
end	NN	O	O
exon	NN	O	O
,	NN	O	O
1B	NN	O	O
.	NN	O	O

FHF2	NN	O	O
is	NN	O	O
a	NN	O	O
large	NN	O	O
gene	NN	O	O
extending	NN	O	O
over	NN	O	O
approximately	NN	O	O
200	NN	O	O
kb	NN	O	O
in	NN	O	O
Xq26	NN	O	O
.	NN	O	O

3	NN	O	O
and	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
seven	NN	O	O
exons	NN	O	O
.	NN	O	O

It	NN	O	O
shows	NN	O	O
tissue	NN	O	O
-	NN	O	O
specific	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
and	NN	O	O
alternative	NN	O	O
transcription	NN	O	O
starts	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
hybridisation	NN	O	O
showed	NN	O	O
highest	NN	O	O
expression	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
.	NN	O	O

The	NN	O	O
FHF2	NN	O	O
gene	NN	O	O
localisation	NN	O	O
and	NN	O	O
tissue	NN	O	O
-	NN	O	O
specific	NN	O	O
expression	NN	O	O
pattern	NN	O	O
suggest	NN	O	O
it	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
familial	NN	O	O
cases	NN	O	O
of	NN	O	O
the	NN	O	O
BFLS	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
other	NN	O	O
syndromal	NN	O	O
and	NN	O	O
non	NN	O	O
-	NN	O	O
specific	NN	O	O
forms	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
mental	NN	O	I-Disease
retardation	NN	O	I-Disease
mapping	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
.	NN	O	O

Germline	NN	O	O
E	NN	O	O
-	NN	O	O
cadherin	NN	O	O
gene	NN	O	O
(	NN	O	O
CDH1	NN	O	O
)	NN	O	O
mutations	NN	O	O
predispose	NN	O	O
to	NN	O	O
familial	NN	O	B-Disease
gastric	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Inherited	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
E	NN	O	O
-	NN	O	O
cadherin	NN	O	O
gene	NN	O	O
(	NN	O	O
CDH1	NN	O	O
)	NN	O	O
were	NN	O	O
described	NN	O	O
recently	NN	O	O
in	NN	O	O
three	NN	O	O
Maori	NN	O	O
kindreds	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
gastric	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Familial	NN	O	B-Disease
gastric	NN	O	I-Disease
cancer	NN	O	I-Disease
is	NN	O	O
genetically	NN	O	O
heterogeneous	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
what	NN	O	O
proportion	NN	O	O
of	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Maori	NN	O	O
populations	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
germline	NN	O	O
CDH1	NN	O	O
mutations	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
screened	NN	O	O
eight	NN	O	O
familial	NN	O	B-Disease
gastric	NN	O	I-Disease
cancer	NN	O	I-Disease
kindreds	NN	O	O
of	NN	O	O
British	NN	O	O
and	NN	O	O
Irish	NN	O	O
origin	NN	O	O
for	NN	O	O
germline	NN	O	O
CDH1	NN	O	O
mutations	NN	O	O
,	NN	O	O
by	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
of	NN	O	O
all	NN	O	O
16	NN	O	O
exons	NN	O	O
and	NN	O	O
flanking	NN	O	O
sequences	NN	O	O
.	NN	O	O

Each	NN	O	O
family	NN	O	O
contained	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
two	NN	O	O
cases	NN	O	O
of	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
in	NN	O	O
first	NN	O	O
degree	NN	O	O
relatives	NN	O	O
with	NN	O	O
one	NN	O	O
affected	NN	O	O
before	NN	O	O
age	NN	O	O
50	NN	O	O
years	NN	O	O
;	NN	O	O
or	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
three	NN	O	O
or	NN	O	O
more	NN	O	O
cases	NN	O	O
of	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Novel	NN	O	O
germline	NN	O	O
CDH1	NN	O	O
mutations	NN	O	O
(	NN	O	O
a	NN	O	O
nonsense	NN	O	O
and	NN	O	O
a	NN	O	O
splice	NN	O	O
site	NN	O	O
)	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
(	NN	O	O
25	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
were	NN	O	O
predicted	NN	O	O
to	NN	O	O
truncate	NN	O	O
the	NN	O	O
E	NN	O	O
-	NN	O	O
cadherin	NN	O	O
protein	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
peptide	NN	O	O
domain	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
family	NN	O	O
there	NN	O	O
was	NN	O	O
evidence	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
penetrance	NN	O	O
and	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
both	NN	O	O
gastric	NN	O	B-Disease
and	NN	O	I-Disease
colorectal	NN	O	I-Disease
cancer	NN	O	I-Disease
;	NN	O	O
thus	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
six	NN	O	O
cases	NN	O	O
of	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
CDH1	NN	O	O
mutation	NN	O	O
carrier	NN	O	O
developed	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
at	NN	O	O
age	NN	O	O
30	NN	O	O
years	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
confirmed	NN	O	O
that	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
CDH1	NN	O	O
gene	NN	O	O
cause	NN	O	O
familial	NN	O	B-Disease
gastric	NN	O	I-Disease
cancer	NN	O	I-Disease
in	NN	O	O
non	NN	O	O
-	NN	O	O
Maori	NN	O	O
populations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
minority	NN	O	O
of	NN	O	O
familial	NN	O	O
gastric	NN	O	B-Disease
cancers	NN	O	I-Disease
can	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
CDH1	NN	O	O
mutations	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
E	NN	O	O
-	NN	O	O
cadherin	NN	O	O
function	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
sporadic	NN	O	O
colorectal	NN	O	B-Disease
and	NN	O	I-Disease
other	NN	O	I-Disease
cancers	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
our	NN	O	O
findings	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
germline	NN	O	O
CDH1	NN	O	O
mutations	NN	O	O
predispose	NN	O	O
to	NN	O	O
early	NN	O	O
onset	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
CDH1	NN	O	O
should	NN	O	O
be	NN	O	O
investigated	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
inherited	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
both	NN	O	O
gastric	NN	O	B-Disease
and	NN	O	I-Disease
colorectal	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
zinc	NN	O	O
finger	NN	O	O
truncation	NN	O	O
of	NN	O	O
murine	NN	O	O
WT1	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
characteristic	NN	O	O
urogenital	NN	O	B-Disease
abnormalities	NN	O	I-Disease
of	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
-	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
WT1	NN	O	O
,	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
urogenital	NN	O	O
development	NN	O	O
,	NN	O	O
and	NN	O	O
WT1	NN	O	B-Disease
dysfunction	NN	O	I-Disease
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
both	NN	O	O
neoplastic	NN	O	B-Disease
(	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
,	NN	O	O
mesothelioma	NN	O	B-Disease
,	NN	O	O
leukemias	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
)	NN	O	O
and	NN	O	O
nonneoplastic	NN	O	B-Disease
(	NN	O	O
glomerulosclerosis	NN	O	B-Disease
)	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
analysis	NN	O	O
of	NN	O	O
diseases	NN	O	O
linked	NN	O	O
specifically	NN	O	O
with	NN	O	O
WT1	NN	O	O
mutations	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
DDS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
can	NN	O	O
provide	NN	O	O
valuable	NN	O	O
insight	NN	O	O
concerning	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
WT1	NN	O	O
in	NN	O	O
development	NN	O	O
and	NN	O	O
disease	NN	O	O
.	NN	O	O

DDS	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
childhood	NN	O	O
disease	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
nephropathy	NN	O	B-Disease
involving	NN	O	O
mesangial	NN	O	B-Disease
sclerosis	NN	O	I-Disease
,	NN	O	O
XY	NN	O	O
pseudohermaphroditism	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
(	NN	O	O
WT	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

DDS	NN	O	B-Disease
patients	NN	O	O
are	NN	O	O
constitutionally	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
exonic	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
WT1	NN	O	O
,	NN	O	O
which	NN	O	O
include	NN	O	O
mutations	NN	O	O
predicted	NN	O	O
to	NN	O	O
truncate	NN	O	O
the	NN	O	O
protein	NN	O	O
within	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
zinc	NN	O	O
finger	NN	O	O
(	NN	O	O
ZF	NN	O	O
)	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
heterozygosity	NN	O	O
for	NN	O	O
a	NN	O	O
targeted	NN	O	O
murine	NN	O	O
Wt1	NN	O	O
allele	NN	O	O
,	NN	O	O
Wt1	NN	O	O
(	NN	O	O
tmT396	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
truncates	NN	O	O
ZF3	NN	O	O
at	NN	O	O
codon	NN	O	O
396	NN	O	O
,	NN	O	O
induces	NN	O	O
mesangial	NN	O	B-Disease
sclerosis	NN	O	I-Disease
characteristic	NN	O	O
of	NN	O	O
DDS	NN	O	B-Disease
in	NN	O	O
adult	NN	O	O
heterozygous	NN	O	O
and	NN	O	O
chimeric	NN	O	O
mice	NN	O	O
.	NN	O	O

Male	NN	O	B-Disease
genital	NN	O	I-Disease
defects	NN	O	I-Disease
also	NN	O	O
were	NN	O	O
evident	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
single	NN	O	O
case	NN	O	O
of	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
in	NN	O	O
which	NN	O	O
the	NN	O	O
transcript	NN	O	O
of	NN	O	O
the	NN	O	O
nontargeted	NN	O	O
allele	NN	O	O
showed	NN	O	O
an	NN	O	O
exon	NN	O	O
9	NN	O	O
skipping	NN	O	O
event	NN	O	O
,	NN	O	O
implying	NN	O	O
a	NN	O	O
causal	NN	O	O
link	NN	O	O
between	NN	O	O
Wt1	NN	O	B-Disease
dysfunction	NN	O	I-Disease
and	NN	O	O
Wilms	NN	O	B-Disease
tumorigenesis	NN	O	I-Disease
in	NN	O	O
mice	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mutant	NN	O	O
WT1	NN	O	O
(	NN	O	O
tmT396	NN	O	O
)	NN	O	O
protein	NN	O	O
accounted	NN	O	O
for	NN	O	O
only	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
WT1	NN	O	O
in	NN	O	O
both	NN	O	O
heterozygous	NN	O	O
embryonic	NN	O	O
stem	NN	O	O
cells	NN	O	O
and	NN	O	O
the	NN	O	O
WT	NN	O	B-Disease
.	NN	O	O

This	NN	O	O
has	NN	O	O
implications	NN	O	O
regarding	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
exerts	NN	O	O
its	NN	O	O
effect	NN	O	O
.	NN	O	O

Mechanism	NN	O	O
of	NN	O	O
increased	NN	O	O
iron	NN	O	O
absorption	NN	O	O
in	NN	O	O
murine	NN	O	O
model	NN	O	O
of	NN	O	O
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
:	NN	O	O
increased	NN	O	O
duodenal	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
iron	NN	O	O
transporter	NN	O	O
DMT1	NN	O	O
.	NN	O	O

Hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
(	NN	O	O
HH	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
tissue	NN	O	O
iron	NN	O	O
deposition	NN	O	O
secondary	NN	O	O
to	NN	O	O
excessive	NN	O	O
dietary	NN	O	O
iron	NN	O	O
absorption	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
HFE	NN	O	O
,	NN	O	O
the	NN	O	O
protein	NN	O	O
defective	NN	O	O
in	NN	O	O
HH	NN	O	B-Disease
,	NN	O	O
was	NN	O	O
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
transferrin	NN	O	O
receptor	NN	O	O
(	NN	O	O
TfR	NN	O	O
)	NN	O	O
in	NN	O	O
duodenal	NN	O	O
crypt	NN	O	O
cells	NN	O	O
and	NN	O	O
proposed	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
HFE	NN	O	O
attenuate	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
transferrin	NN	O	O
-	NN	O	O
bound	NN	O	O
iron	NN	O	O
from	NN	O	O
plasma	NN	O	O
by	NN	O	O
duodenal	NN	O	O
crypt	NN	O	O
cells	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
up	NN	O	O
-	NN	O	O
regulation	NN	O	O
of	NN	O	O
transporters	NN	O	O
for	NN	O	O
dietary	NN	O	O
iron	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
HFE	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
have	NN	O	O
increased	NN	O	O
duodenal	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
divalent	NN	O	O
metal	NN	O	O
transporter	NN	O	O
(	NN	O	O
DMT1	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
4	NN	O	O
weeks	NN	O	O
of	NN	O	O
age	NN	O	O
,	NN	O	O
the	NN	O	O
HFE	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
demonstrated	NN	O	O
iron	NN	O	O
loading	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
HFE	NN	O	O
+	NN	O	O
/	NN	O	O
+	NN	O	O
littermates	NN	O	O
,	NN	O	O
with	NN	O	O
elevated	NN	O	O
transferrin	NN	O	O
saturations	NN	O	O
(	NN	O	O
68	NN	O	O
.	NN	O	O
4	NN	O	O
%	NN	O	O
vs	NN	O	O
.	NN	O	O
49	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
elevated	NN	O	O
liver	NN	O	O
iron	NN	O	O
concentrations	NN	O	O
(	NN	O	O
985	NN	O	O
micrograms	NN	O	O
vs	NN	O	O
.	NN	O	O
381	NN	O	O
micrograms	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
quantitated	NN	O	O
duodenal	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
classes	NN	O	O
of	NN	O	O
DMT1	NN	O	O
transcripts	NN	O	O
one	NN	O	O
containing	NN	O	O
an	NN	O	O
iron	NN	O	O
responsive	NN	O	O
element	NN	O	O
(	NN	O	O
IRE	NN	O	O
)	NN	O	O
,	NN	O	O
called	NN	O	O
DMT1	NN	O	O
(	NN	O	O
IRE	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
containing	NN	O	O
no	NN	O	O
IRE	NN	O	O
,	NN	O	O
called	NN	O	O
DMT1	NN	O	O
(	NN	O	O
non	NN	O	O
-	NN	O	O
IRE	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
positive	NN	O	O
control	NN	O	O
for	NN	O	O
DMT1	NN	O	O
up	NN	O	O
-	NN	O	O
regulation	NN	O	O
was	NN	O	O
a	NN	O	O
murine	NN	O	O
model	NN	O	O
of	NN	O	O
dietary	NN	O	B-Disease
iron	NN	O	I-Disease
deficiency	NN	O	I-Disease
that	NN	O	O
demonstrated	NN	O	O
greatly	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
duodenal	NN	O	O
DMT1	NN	O	O
(	NN	O	O
IRE	NN	O	O
)	NN	O	O
mRNA	NN	O	O
.	NN	O	O

HFE	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
duodenal	NN	O	O
DMT1	NN	O	O
(	NN	O	O
IRE	NN	O	O
)	NN	O	O
mRNA	NN	O	O
(	NN	O	O
average	NN	O	O
7	NN	O	O
.	NN	O	O
7	NN	O	O
-	NN	O	O
fold	NN	O	O
)	NN	O	O
,	NN	O	O
despite	NN	O	O
their	NN	O	O
elevated	NN	O	O
transferrin	NN	O	O
saturation	NN	O	O
and	NN	O	O
hepatic	NN	O	O
iron	NN	O	O
content	NN	O	O
.	NN	O	O

Duodenal	NN	O	O
expression	NN	O	O
of	NN	O	O
DMT1	NN	O	O
(	NN	O	O
non	NN	O	O
-	NN	O	O
IRE	NN	O	O
)	NN	O	O
was	NN	O	O
not	NN	O	O
increased	NN	O	O
,	NN	O	O
nor	NN	O	O
was	NN	O	O
hepatic	NN	O	O
expression	NN	O	O
of	NN	O	O
DMT1	NN	O	O
increased	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
model	NN	O	O
for	NN	O	O
HH	NN	O	O
in	NN	O	O
which	NN	O	O
HFE	NN	O	O
mutations	NN	O	O
lead	NN	O	O
to	NN	O	O
inappropriately	NN	O	O
low	NN	O	O
crypt	NN	O	O
cell	NN	O	O
iron	NN	O	O
,	NN	O	O
with	NN	O	O
resultant	NN	O	O
stabilization	NN	O	O
of	NN	O	O
DMT1	NN	O	O
(	NN	O	O
IRE	NN	O	O
)	NN	O	O
mRNA	NN	O	O
,	NN	O	O
up	NN	O	O
-	NN	O	O
regulation	NN	O	O
of	NN	O	O
DMT1	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
absorption	NN	O	O
of	NN	O	O
dietary	NN	O	O
iron	NN	O	O
.	NN	O	O

Neurophysiologic	NN	O	O
follow	NN	O	O
-	NN	O	O
up	NN	O	O
of	NN	O	O
long	NN	O	O
-	NN	O	O
term	NN	O	O
dietary	NN	O	O
treatment	NN	O	O
in	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
.	NN	O	O

OBJECTIVE	NN	O	O
To	NN	O	O
monitor	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
dietary	NN	O	O
treatment	NN	O	O
in	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
somatosensory	NN	O	O
evoked	NN	O	O
potentials	NN	O	O
(	NN	O	O
SEPs	NN	O	O
)	NN	O	O
and	NN	O	O
motor	NN	O	O
evoked	NN	O	O
potentials	NN	O	O
(	NN	O	O
MEPs	NN	O	O
)	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
SEPs	NN	O	O
and	NN	O	O
MEPs	NN	O	O
have	NN	O	O
proved	NN	O	O
useful	NN	O	O
in	NN	O	O
revealing	NN	O	O
signs	NN	O	O
of	NN	O	O
progressively	NN	O	O
severe	NN	O	O
,	NN	O	O
central	NN	O	O
dying	NN	O	B-Disease
-	NN	O	I-Disease
back	NN	O	I-Disease
axonopathy	NN	O	I-Disease
in	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

METHODS	NN	O	O
Eight	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
ALD	NN	O	B-Disease
underwent	NN	O	O
clinical	NN	O	O
examination	NN	O	O
,	NN	O	O
brain	NN	O	O
and	NN	O	O
spine	NN	O	O
MRI	NN	O	O
,	NN	O	O
and	NN	O	O
SEP	NN	O	O
and	NN	O	O
MEP	NN	O	O
studies	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
3	NN	O	O
years	NN	O	O
of	NN	O	O
Lorenzos	NN	O	O
oil	NN	O	O
dietary	NN	O	O
therapy	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Before	NN	O	O
treatment	NN	O	O
,	NN	O	O
brain	NN	O	O
MRI	NN	O	O
was	NN	O	O
normal	NN	O	O
in	NN	O	O
five	NN	O	O
patients	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
had	NN	O	O
pure	NN	O	O
spinal	NN	O	B-Disease
SEP	NN	O	I-Disease
abnormalities	NN	O	I-Disease
and	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
two	NN	O	O
patients	NN	O	O
SEPs	NN	O	O
showed	NN	O	O
signs	NN	O	O
of	NN	O	O
involvement	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
spinal	NN	O	O
and	NN	O	O
cerebral	NN	O	O
somatosensory	NN	O	O
tracts	NN	O	O
.	NN	O	O

After	NN	O	O
treatment	NN	O	O
,	NN	O	O
the	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
pure	NN	O	O
spinal	NN	O	B-Disease
abnormalities	NN	O	I-Disease
showed	NN	O	O
clinical	NN	O	O
and	NN	O	O
neurophysiologic	NN	O	O
worsening	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
more	NN	O	O
advanced	NN	O	O
stage	NN	O	O
of	NN	O	O
disease	NN	O	O
(	NN	O	O
exhibited	NN	O	O
by	NN	O	O
SEPs	NN	O	O
)	NN	O	O
showed	NN	O	O
substantially	NN	O	O
unchanged	NN	O	O
clinical	NN	O	O
and	NN	O	O
neurophysiologic	NN	O	O
features	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
with	NN	O	O
abnormal	NN	O	O
brain	NN	O	O
MRI	NN	O	O
at	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
treatment	NN	O	O
showed	NN	O	O
clinical	NN	O	O
and	NN	O	O
neurophysiologic	NN	O	O
worsening	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
Lorenzos	NN	O	O
oil	NN	O	O
therapy	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
patients	NN	O	O
with	NN	O	O
evidence	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-Disease
brain	NN	O	I-Disease
lesions	NN	O	I-Disease
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
patients	NN	O	O
without	NN	O	O
clear	NN	O	O
signs	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
damage	NN	O	O
,	NN	O	O
this	NN	O	O
treatment	NN	O	O
does	NN	O	O
not	NN	O	O
modify	NN	O	O
significantly	NN	O	O
the	NN	O	O
natural	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
because	NN	O	O
effective	NN	O	O
treatments	NN	O	O
should	NN	O	O
begin	NN	O	O
before	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
severe	NN	O	O
neurologic	NN	O	O
symptoms	NN	O	O
,	NN	O	O
SEPs	NN	O	O
and	NN	O	O
MEPs	NN	O	O
should	NN	O	O
be	NN	O	O
considered	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
other	NN	O	O
experimental	NN	O	O
treatments	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
negative	NN	O	O
brain	NN	O	O
MRI	NN	O	O
.	NN	O	O

GCH1	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
oromandibular	NN	O	B-Disease
dystonia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
authors	NN	O	O
report	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
of	NN	O	O
GCH1	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
oromandibular	NN	O	B-Disease
dystonia	NN	O	I-Disease
and	NN	O	O
no	NN	O	O
obvious	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
dystonia	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
patient	NN	O	O
responded	NN	O	O
positively	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
L	NN	O	O
-	NN	O	O
dopa	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
GCH1	NN	O	O
mutations	NN	O	O
must	NN	O	O
be	NN	O	O
considered	NN	O	O
even	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
dystonic	NN	O	B-Disease
symptoms	NN	O	O
not	NN	O	O
typical	NN	O	O
of	NN	O	O
dopa	NN	O	B-Disease
-	NN	O	I-Disease
responsive	NN	O	I-Disease
dystonia	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
Korean	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

We	NN	O	O
extensively	NN	O	O
analyzed	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
and	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
(	NN	O	O
mRNA	NN	O	O
)	NN	O	O
from	NN	O	O
62	NN	O	O
unrelated	NN	O	O
Korean	NN	O	O
patients	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
for	NN	O	O
identification	NN	O	O
of	NN	O	O
germline	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
adopted	NN	O	O
both	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
and	NN	O	O
a	NN	O	O
method	NN	O	O
of	NN	O	O
analysis	NN	O	O
involving	NN	O	O
the	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
-	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
)	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
(	NN	O	O
PTT	NN	O	O
)	NN	O	O
.	NN	O	O

DNA	NN	O	O
sequencing	NN	O	O
confirmed	NN	O	O
all	NN	O	O
alterations	NN	O	O
represented	NN	O	O
by	NN	O	O
aberrant	NN	O	O
bands	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
38	NN	O	O
patients	NN	O	O
(	NN	O	O
61	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Nineteen	NN	O	O
of	NN	O	O
the	NN	O	O
detected	NN	O	O
mutations	NN	O	O
were	NN	O	O
presumed	NN	O	O
to	NN	O	O
be	NN	O	O
novel	NN	O	O
,	NN	O	O
thus	NN	O	O
emphasizing	NN	O	O
the	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
mutational	NN	O	O
spectrum	NN	O	O
in	NN	O	O
Korean	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
initial	NN	O	O
48	NN	O	O
patients	NN	O	O
,	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
PTT	NN	O	O
for	NN	O	O
those	NN	O	O
patients	NN	O	O
for	NN	O	O
whom	NN	O	O
no	NN	O	O
detectable	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
by	NN	O	O
SSCP	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
combined	NN	O	O
approach	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
germline	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
mutations	NN	O	O
in	NN	O	O
29	NN	O	O
of	NN	O	O
the	NN	O	O
48	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
(	NN	O	O
60	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
including	NN	O	O
6	NN	O	O
patients	NN	O	O
in	NN	O	O
whom	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
failed	NN	O	O
to	NN	O	O
distinguish	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
14	NN	O	O
later	NN	O	O
patients	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
in	NN	O	O
9	NN	O	O
patients	NN	O	O
(	NN	O	O
64	NN	O	O
%	NN	O	O
)	NN	O	O
using	NN	O	O
PTT	NN	O	O
only	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
confirm	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
detection	NN	O	O
rate	NN	O	O
with	NN	O	O
PTT	NN	O	O
was	NN	O	O
superior	NN	O	O
to	NN	O	O
that	NN	O	O
with	NN	O	O
SSCP	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
PTT	NN	O	O
would	NN	O	O
be	NN	O	O
a	NN	O	O
more	NN	O	O
practical	NN	O	O
screening	NN	O	O
method	NN	O	O
to	NN	O	O
detect	NN	O	O
germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Molecular	NN	O	O
epidemiology	NN	O	O
of	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
heterozygotes	NN	O	O
with	NN	O	O
an	NN	O	O
Arg95Stop	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
C9	NN	O	O
gene	NN	O	O
in	NN	O	O
Japan	NN	O	O
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
ninth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
human	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C9	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
complement	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Japan	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
incidence	NN	O	O
of	NN	O	O
approximately	NN	O	O
one	NN	O	O
homozygote	NN	O	O
in	NN	O	O
1000	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
very	NN	O	O
rare	NN	O	O
in	NN	O	O
other	NN	O	O
countries	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analyses	NN	O	O
of	NN	O	O
Japanese	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
have	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
leading	NN	O	O
to	NN	O	O
TGA	NN	O	O
stop	NN	O	O
codon	NN	O	O
for	NN	O	O
Arg95	NN	O	O
in	NN	O	O
exon	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
C9	NN	O	O
gene	NN	O	O
(	NN	O	O
Arg95Stop	NN	O	O
)	NN	O	O
is	NN	O	O
common	NN	O	O
in	NN	O	O
Japanese	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
heterozygous	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
Arg95Stop	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
population	NN	O	O
,	NN	O	O
we	NN	O	O
collected	NN	O	O
DNA	NN	O	O
samples	NN	O	O
from	NN	O	O
300	NN	O	O
individuals	NN	O	O
in	NN	O	O
two	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
main	NN	O	O
islands	NN	O	O
of	NN	O	O
Japan	NN	O	O
.	NN	O	O

Heterozygote	NN	O	O
detection	NN	O	O
was	NN	O	O
performed	NN	O	O
with	NN	O	O
an	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
system	NN	O	O
designed	NN	O	O
to	NN	O	O
detect	NN	O	O
exclusively	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
confirmation	NN	O	O
with	NN	O	O
PCR	NN	O	O
/	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Twenty	NN	O	O
individuals	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
Arg95Stop	NN	O	O
mutation	NN	O	O
.	NN	O	O

None	NN	O	O
was	NN	O	O
homozygous	NN	O	O
.	NN	O	O

The	NN	O	O
prevalence	NN	O	O
of	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
Arg95Stop	NN	O	O
mutation	NN	O	O
was	NN	O	O
6	NN	O	O
.	NN	O	O

7	NN	O	O
%	NN	O	O
(	NN	O	O
20	NN	O	O
/	NN	O	O
300	NN	O	O
)	NN	O	O
.	NN	O	O

An	NN	O	O
estimated	NN	O	O
frequency	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
12	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
complete	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
homozygous	NN	O	O
Arg95Stop	NN	O	O
mutation	NN	O	O
was	NN	O	O
consistent	NN	O	O
with	NN	O	O
frequencies	NN	O	O
determined	NN	O	O
by	NN	O	O
serological	NN	O	O
studies	NN	O	O

The	NN	O	O
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
protein	NN	O	O
,	NN	O	O
HFE	NN	O	O
,	NN	O	O
specifically	NN	O	O
regulates	NN	O	O
transferrin	NN	O	O
-	NN	O	O
mediated	NN	O	O
iron	NN	O	O
uptake	NN	O	O
in	NN	O	O
HeLa	NN	O	O
cells	NN	O	O
.	NN	O	O

HFE	NN	O	O
is	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
mutated	NN	O	O
in	NN	O	O
the	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
(	NN	O	O
Feder	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O

N	NN	O	O
.	NN	O	O
,	NN	O	O
Gnirke	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Thomas	NN	O	O
,	NN	O	O
W	NN	O	O
.	NN	O	O
,	NN	O	O
Tsuchihashi	NN	O	O
,	NN	O	O
Z	NN	O	O
.	NN	O	O
,	NN	O	O
Ruddy	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O

A	NN	O	O
.	NN	O	O
,	NN	O	O
Basava	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Dormishian	NN	O	O
,	NN	O	O
F	NN	O	O
.	NN	O	O
,	NN	O	O
Domingo	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O

J	NN	O	O
.	NN	O	O
,	NN	O	O
Ellis	NN	O	O
,	NN	O	O
M	NN	O	O
.	NN	O	O

C	NN	O	O
.	NN	O	O
,	NN	O	O
Fullan	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Hinton	NN	O	O
,	NN	O	O
L	NN	O	O
.	NN	O	O

M	NN	O	O
.	NN	O	O
,	NN	O	O
Jones	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O

L	NN	O	O
.	NN	O	O
,	NN	O	O
Kimmel	NN	O	O
,	NN	O	O
B	NN	O	O
.	NN	O	O

E	NN	O	O
.	NN	O	O
,	NN	O	O
Kronmal	NN	O	O
,	NN	O	O
G	NN	O	O
.	NN	O	O

S	NN	O	O
.	NN	O	O
,	NN	O	O
Lauer	NN	O	O
,	NN	O	O
P	NN	O	O
.	NN	O	O
,	NN	O	O
Lee	NN	O	O
,	NN	O	O
V	NN	O	O
.	NN	O	O

K	NN	O	O
.	NN	O	O
,	NN	O	O
Loeb	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O

B	NN	O	O
.	NN	O	O
,	NN	O	O
Mapa	NN	O	O
,	NN	O	O
F	NN	O	O
.	NN	O	O

A	NN	O	O
.	NN	O	O
,	NN	O	O
McClelland	NN	O	O
,	NN	O	O
E	NN	O	O
.	NN	O	O
,	NN	O	O
Meyer	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O

C	NN	O	O
.	NN	O	O
,	NN	O	O
Mintier	NN	O	O
,	NN	O	O
G	NN	O	O
.	NN	O	O

A	NN	O	O
.	NN	O	O
,	NN	O	O
Moeller	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O
,	NN	O	O
Moore	NN	O	O
,	NN	O	O
T	NN	O	O
.	NN	O	O
,	NN	O	O
Morikang	NN	O	O
,	NN	O	O
E	NN	O	O
.	NN	O	O
,	NN	O	O
Prasss	NN	O	O
,	NN	O	O
C	NN	O	O
.	NN	O	O

E	NN	O	O
.	NN	O	O
,	NN	O	O
Quintana	NN	O	O
,	NN	O	O
L	NN	O	O
.	NN	O	O
,	NN	O	O
Starnes	NN	O	O
,	NN	O	O
S	NN	O	O
.	NN	O	O

M	NN	O	O
.	NN	O	O
,	NN	O	O
Schatzman	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O

C	NN	O	O
.	NN	O	O
,	NN	O	O
Brunke	NN	O	O
,	NN	O	O
K	NN	O	O
.	NN	O	O

J	NN	O	O
.	NN	O	O
,	NN	O	O
Drayna	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O

T	NN	O	O
.	NN	O	O
,	NN	O	O
Risch	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O

J	NN	O	O
.	NN	O	O
,	NN	O	O
Bacon	NN	O	O
,	NN	O	O
B	NN	O	O
.	NN	O	O

R	NN	O	O
.	NN	O	O
,	NN	O	O
and	NN	O	O
Wolff	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O

R	NN	O	O
.	NN	O	O

(	NN	O	O
1996	NN	O	O
)	NN	O	O
Nat	NN	O	O
.	NN	O	O

Genet	NN	O	O
.	NN	O	O

13	NN	O	O
,	NN	O	O
399	NN	O	O
-	NN	O	O
408	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
,	NN	O	O
HFE	NN	O	O
complexes	NN	O	O
with	NN	O	O
transferrin	NN	O	O
receptor	NN	O	O
(	NN	O	O
TfR	NN	O	O
)	NN	O	O
,	NN	O	O
increasing	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
transferrin	NN	O	O
(	NN	O	O
Tf	NN	O	O
)	NN	O	O
for	NN	O	O
its	NN	O	O
receptor	NN	O	O
10	NN	O	O
-	NN	O	O
fold	NN	O	O
(	NN	O	O
Gross	NN	O	O
,	NN	O	O
C	NN	O	O
.	NN	O	O

N	NN	O	O
.	NN	O	O
,	NN	O	O
Irrinki	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Feder	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O
N	NN	O	O
.	NN	O	O
,	NN	O	O
and	NN	O	O
Enns	NN	O	O
,	NN	O	O
C	NN	O	O
.	NN	O	O
A	NN	O	O
.	NN	O	O
(	NN	O	O
1998	NN	O	O
)	NN	O	O
J	NN	O	O
.	NN	O	O
Biol	NN	O	O
.	NN	O	O
Chem	NN	O	O
.	NN	O	O
273	NN	O	O
,	NN	O	O
22068	NN	O	O
-	NN	O	O
22074	NN	O	O
;	NN	O	O
Feder	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O
N	NN	O	O
.	NN	O	O
,	NN	O	O
Penny	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O
M	NN	O	O
.	NN	O	O
,	NN	O	O
Irrinki	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Lee	NN	O	O
,	NN	O	O
V	NN	O	O
.	NN	O	O
K	NN	O	O
.	NN	O	O
,	NN	O	O
Lebron	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O
A	NN	O	O
.	NN	O	O
,	NN	O	O
Watson	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O
,	NN	O	O
Tsuchihashi	NN	O	O
,	NN	O	O
Z	NN	O	O
.	NN	O	O
,	NN	O	O
Sigal	NN	O	O
,	NN	O	O
E	NN	O	O
.	NN	O	O
,	NN	O	O
Bjorkman	NN	O	O
,	NN	O	O
P	NN	O	O
.	NN	O	O
J	NN	O	O
.	NN	O	O
,	NN	O	O
and	NN	O	O
Schatzman	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O
C	NN	O	O
.	NN	O	O
(	NN	O	O
1998	NN	O	O
)	NN	O	O
Proc	NN	O	O
.	NN	O	O
Natl	NN	O	O
.	NN	O	O

Acad	NN	O	O
.	NN	O	O

Sci	NN	O	O
.	NN	O	O

U	NN	O	O
S	NN	O	O
A	NN	O	O
95	NN	O	O
,	NN	O	O
1472	NN	O	O
-	NN	O	O
1477	NN	O	O
)	NN	O	O
.	NN	O	O

HFE	NN	O	O
does	NN	O	O
not	NN	O	O
remain	NN	O	O
at	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
,	NN	O	O
but	NN	O	O
traffics	NN	O	O
with	NN	O	O
TfR	NN	O	O
to	NN	O	O
Tf	NN	O	O
-	NN	O	O
positive	NN	O	O
internal	NN	O	O
compartments	NN	O	O
(	NN	O	O
Gross	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1998	NN	O	O
)	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
HeLa	NN	O	O
cell	NN	O	O
line	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
HFE	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
tetracycline	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
HFE	NN	O	O
reduces	NN	O	O
55Fe	NN	O	O
uptake	NN	O	O
from	NN	O	O
Tf	NN	O	O
by	NN	O	O
33	NN	O	O
%	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
endocytic	NN	O	O
or	NN	O	O
exocytic	NN	O	O
rates	NN	O	O
of	NN	O	O
TfR	NN	O	O
cycling	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
HFE	NN	O	O
appears	NN	O	O
to	NN	O	O
reduce	NN	O	O
cellular	NN	O	O
acquisition	NN	O	O
of	NN	O	O
iron	NN	O	O
from	NN	O	O
Tf	NN	O	O
within	NN	O	O
endocytic	NN	O	O
compartments	NN	O	O
.	NN	O	O

HFE	NN	O	O
specifically	NN	O	O
reduces	NN	O	O
iron	NN	O	O
uptake	NN	O	O
from	NN	O	O
Tf	NN	O	O
,	NN	O	O
as	NN	O	O
non	NN	O	O
-	NN	O	O
Tf	NN	O	O
-	NN	O	O
mediated	NN	O	O
iron	NN	O	O
uptake	NN	O	O
from	NN	O	O
Fe	NN	O	O
-	NN	O	O
nitrilotriacetic	NN	O	O
acid	NN	O	O
is	NN	O	O
not	NN	O	O
altered	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
explain	NN	O	O
the	NN	O	O
decreased	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
seen	NN	O	O
in	NN	O	O
our	NN	O	O
HeLa	NN	O	O
cell	NN	O	O
system	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
specific	NN	O	O
control	NN	O	O
of	NN	O	O
HFE	NN	O	O
over	NN	O	O
the	NN	O	O
Tf	NN	O	O
-	NN	O	O
mediated	NN	O	O
pathway	NN	O	O
of	NN	O	O
iron	NN	O	O
uptake	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
also	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
cellular	NN	O	O
iron	NN	O	O
homeostasis	NN	O	O
in	NN	O	O
organs	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
liver	NN	O	O
,	NN	O	O
pancreas	NN	O	O
,	NN	O	O
heart	NN	O	O
,	NN	O	O
and	NN	O	O
spleen	NN	O	O
that	NN	O	O
are	NN	O	O
iron	NN	O	O
loaded	NN	O	O
in	NN	O	O
hereditary	NN	O	B-Disease
hemochromatotic	NN	O	I-Disease
individuals	NN	O	O
lacking	NN	O	O
functional	NN	O	O
HFE	NN	O	O
.	NN	O	O

Mutation	NN	O	O
and	NN	O	O
haplotype	NN	O	O
studies	NN	O	O
of	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
reveal	NN	O	O
new	NN	O	O
ancestral	NN	O	O
relationships	NN	O	O
and	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
high	NN	O	O
carrier	NN	O	O
frequency	NN	O	O
with	NN	O	O
reduced	NN	O	O
penetrance	NN	O	O
in	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
population	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
recessive	NN	O	B-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
episodes	NN	O	O
of	NN	O	O
fever	NN	O	O
with	NN	O	O
serositis	NN	O	B-Disease
or	NN	O	O
synovitis	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
FMF	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
MEFV	NN	O	O
)	NN	O	O
was	NN	O	O
cloned	NN	O	O
recently	NN	O	O
,	NN	O	O
and	NN	O	O
four	NN	O	O
missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
present	NN	O	O
data	NN	O	O
from	NN	O	O
non	NN	O	O
-	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
and	NN	O	O
Arab	NN	O	O
patients	NN	O	O
in	NN	O	O
whom	NN	O	O
we	NN	O	O
had	NN	O	O
not	NN	O	O
originally	NN	O	O
found	NN	O	O
mutations	NN	O	O
and	NN	O	O
from	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
more	NN	O	O
ethnically	NN	O	O
diverse	NN	O	O
panel	NN	O	O
.	NN	O	O

Among	NN	O	O
90	NN	O	O
symptomatic	NN	O	O
mutation	NN	O	O
-	NN	O	O
positive	NN	O	O
individuals	NN	O	O
,	NN	O	O
11	NN	O	O
mutations	NN	O	O
accounted	NN	O	O
for	NN	O	O
79	NN	O	O
%	NN	O	O
of	NN	O	O
carrier	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
that	NN	O	O
are	NN	O	O
novel	NN	O	O
,	NN	O	O
one	NN	O	O
alters	NN	O	O
the	NN	O	O
same	NN	O	O
residue	NN	O	O
(	NN	O	O
680	NN	O	O
)	NN	O	O
as	NN	O	O
a	NN	O	O
previously	NN	O	O
known	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
(	NN	O	O
P369S	NN	O	O
)	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
another	NN	O	O
recent	NN	O	O
report	NN	O	O
,	NN	O	O
the	NN	O	O
E148Q	NN	O	O
mutation	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
patients	NN	O	O
of	NN	O	O
several	NN	O	O
ethnicities	NN	O	O
and	NN	O	O
on	NN	O	O
multiple	NN	O	O
microsatellite	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
but	NN	O	O
haplotype	NN	O	O
data	NN	O	O
indicate	NN	O	O
an	NN	O	O
ancestral	NN	O	O
relationships	NN	O	O
between	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Italian	NN	O	O
and	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
patients	NN	O	O
with	NN	O	O
FMF	NN	O	B-Disease
and	NN	O	O
other	NN	O	O
affected	NN	O	O
populations	NN	O	O
.	NN	O	O

Among	NN	O	O
approximately	NN	O	O
200	NN	O	O
anonymous	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
DNA	NN	O	O
samples	NN	O	O
,	NN	O	O
the	NN	O	O
MEFV	NN	O	O
carrier	NN	O	O
frequency	NN	O	O
was	NN	O	O
21	NN	O	O
%	NN	O	O
,	NN	O	O
with	NN	O	O
E148Q	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
.	NN	O	O

Several	NN	O	O
lines	NN	O	O
of	NN	O	O
evidence	NN	O	O
indicate	NN	O	O
reduced	NN	O	O
penetrance	NN	O	O
among	NN	O	O
Ashkenazi	NN	O	O
Jews	NN	O	O
,	NN	O	O
especially	NN	O	O
for	NN	O	O
E148Q	NN	O	O
,	NN	O	O
P369S	NN	O	O
,	NN	O	O
and	NN	O	O
K695R	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
E148Q	NN	O	O
helps	NN	O	O
account	NN	O	O
for	NN	O	O
recessive	NN	O	O
inheritance	NN	O	O
in	NN	O	O
an	NN	O	O
Ashkenazi	NN	O	O
family	NN	O	O
previously	NN	O	O
reported	NN	O	O
as	NN	O	O
an	NN	O	O
unusual	NN	O	O
case	NN	O	O
of	NN	O	O
dominantly	NN	O	O
inherited	NN	O	O
FMF	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
three	NN	O	O
frequent	NN	O	O
MEFV	NN	O	O
mutations	NN	O	O
in	NN	O	O
multiple	NN	O	O
Mediterranean	NN	O	O
populations	NN	O	O
strongly	NN	O	O
suggests	NN	O	O
a	NN	O	O
heterozygote	NN	O	O
advantage	NN	O	O
in	NN	O	O
this	NN	O	O
geographic	NN	O	O
region	NN	O	O
.	NN	O	O

Autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
with	NN	O	O
defective	NN	O	O
Fas	NN	O	O
:	NN	O	O
genotype	NN	O	O
influences	NN	O	O
penetrance	NN	O	O
.	NN	O	O

Autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ALPS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
disorder	NN	O	B-Disease
of	NN	O	I-Disease
lymphocyte	NN	O	I-Disease
homeostasis	NN	O	I-Disease
and	NN	O	I-Disease
immunological	NN	O	I-Disease
tolerance	NN	O	I-Disease
.	NN	O	O

Most	NN	O	O
patients	NN	O	O
have	NN	O	O
a	NN	O	O
heterozygous	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
APT1	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
Fas	NN	O	O
(	NN	O	O
CD95	NN	O	O
,	NN	O	O
APO	NN	O	O
-	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
mediator	NN	O	O
of	NN	O	O
an	NN	O	O
apoptotic	NN	O	O
pathway	NN	O	O
crucial	NN	O	O
to	NN	O	O
lymphocyte	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Of	NN	O	O
17	NN	O	O
unique	NN	O	O
APT1	NN	O	O
mutations	NN	O	O
in	NN	O	O
unrelated	NN	O	O
ALPS	NN	O	B-Disease
probands	NN	O	O
,	NN	O	O
12	NN	O	O
(	NN	O	O
71	NN	O	O
%	NN	O	O
)	NN	O	O
occurred	NN	O	O
in	NN	O	O
exons	NN	O	O
7	NN	O	O
-	NN	O	O
9	NN	O	O
,	NN	O	O
which	NN	O	O
encode	NN	O	O
the	NN	O	O
intracellular	NN	O	O
portion	NN	O	O
of	NN	O	O
Fas	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
all	NN	O	O
17	NN	O	O
patients	NN	O	O
showed	NN	O	O
apoptotic	NN	O	O
defects	NN	O	O
when	NN	O	O
exposed	NN	O	O
to	NN	O	O
an	NN	O	O
anti	NN	O	O
-	NN	O	O
Fas	NN	O	O
agonist	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
.	NN	O	O

Similar	NN	O	O
defects	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
Fas	NN	O	O
-	NN	O	O
negative	NN	O	O
cell	NN	O	O
line	NN	O	O
transfected	NN	O	O
with	NN	O	O
cDNAs	NN	O	O
bearing	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
Fas	NN	O	O
constructs	NN	O	O
with	NN	O	O
either	NN	O	O
intra	NN	O	O
-	NN	O	O
or	NN	O	O
extracellular	NN	O	O
mutations	NN	O	O
caused	NN	O	O
dominant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
mediated	NN	O	O
by	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
Fas	NN	O	O
.	NN	O	O

Two	NN	O	O
missense	NN	O	O
Fas	NN	O	O
variants	NN	O	O
,	NN	O	O
not	NN	O	O
restricted	NN	O	O
to	NN	O	O
patients	NN	O	O
with	NN	O	O
ALPS	NN	O	B-Disease
,	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Variant	NN	O	O
A	NN	O	O
(	NN	O	O
-	NN	O	O
1	NN	O	O
)	NN	O	O
T	NN	O	O
at	NN	O	O
the	NN	O	O
Fas	NN	O	O
signal	NN	O	O
-	NN	O	O
sequence	NN	O	O
cleavage	NN	O	O
site	NN	O	O
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
apoptosis	NN	O	O
less	NN	O	O
well	NN	O	O
than	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
Fas	NN	O	O
and	NN	O	O
is	NN	O	O
partially	NN	O	O
inhibitory	NN	O	O
,	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
13	NN	O	O
%	NN	O	O
of	NN	O	O
African	NN	O	O
American	NN	O	O
alleles	NN	O	O
.	NN	O	O

Among	NN	O	O
the	NN	O	O
ALPS	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
Fas	NN	O	O
mutants	NN	O	O
,	NN	O	O
dominant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
much	NN	O	O
more	NN	O	O
pronounced	NN	O	O
in	NN	O	O
mutants	NN	O	O
affecting	NN	O	O
the	NN	O	O
intracellular	NN	O	O
,	NN	O	O
versus	NN	O	O
extracellular	NN	O	O
,	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
Fas	NN	O	O
receptor	NN	O	O
.	NN	O	O

Mutations	NN	O	O
causing	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	O
Fas	NN	O	O
death	NN	O	O
domain	NN	O	O
also	NN	O	O
showed	NN	O	O
a	NN	O	O
higher	NN	O	O
penetrance	NN	O	O
of	NN	O	O
ALPS	NN	O	B-Disease
phenotype	NN	O	O
features	NN	O	O
in	NN	O	O
mutation	NN	O	O
-	NN	O	O
bearing	NN	O	O
relatives	NN	O	O
.	NN	O	O

Significant	NN	O	O
ALPS	NN	O	B-Disease
-	NN	O	O
related	NN	O	O
morbidity	NN	O	O
occurred	NN	O	O
in	NN	O	O
44	NN	O	O
%	NN	O	O
of	NN	O	O
relatives	NN	O	O
with	NN	O	O
intracellular	NN	O	O
mutations	NN	O	O
,	NN	O	O
versus	NN	O	O
0	NN	O	O
%	NN	O	O
of	NN	O	O
relatives	NN	O	O
with	NN	O	O
extracellular	NN	O	O
mutations	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
mutations	NN	O	O
within	NN	O	O
APT1	NN	O	O
strongly	NN	O	O
influences	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
the	NN	O	O
severity	NN	O	O
of	NN	O	O
ALPS	NN	O	B-Disease
.	NN	O	O

Multicentric	NN	O	O
origin	NN	O	O
of	NN	O	O
hemochromatosis	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
HFE	NN	O	O
)	NN	O	O
mutations	NN	O	O
.	NN	O	O

Genetic	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
(	NN	O	O
GH	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
disease	NN	O	O
restricted	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
European	NN	O	O
ancestry	NN	O	O
.	NN	O	O

In	NN	O	O
northwestern	NN	O	O
Europe	NN	O	O
,	NN	O	O
>	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
GH	NN	O	B-Disease
patients	NN	O	O
are	NN	O	O
homozygous	NN	O	O
for	NN	O	O
one	NN	O	O
mutation	NN	O	O
,	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
for	NN	O	O
cysteine	NN	O	O
at	NN	O	O
position	NN	O	O
282	NN	O	O
(	NN	O	O
C282Y	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
unprocessed	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
proportion	NN	O	O
of	NN	O	O
GH	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
two	NN	O	O
mutations	NN	O	O
are	NN	O	O
present	NN	O	O
,	NN	O	O
C282Y	NN	O	O
and	NN	O	O
H63D	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
second	NN	O	O
mutation	NN	O	O
is	NN	O	O
such	NN	O	O
that	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
predispose	NN	O	O
1	NN	O	O
%	NN	O	O
-	NN	O	O
2	NN	O	O
%	NN	O	O
of	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
differ	NN	O	O
,	NN	O	O
C282Y	NN	O	O
being	NN	O	O
limited	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
northwestern	NN	O	O
European	NN	O	O
ancestry	NN	O	O
and	NN	O	O
H63D	NN	O	O
being	NN	O	O
found	NN	O	O
at	NN	O	O
allele	NN	O	O
frequencies	NN	O	O
>	NN	O	O
5	NN	O	O
%	NN	O	O
,	NN	O	O
in	NN	O	O
Europe	NN	O	O
,	NN	O	O
in	NN	O	O
countries	NN	O	O
bordering	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
Middle	NN	O	O
East	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
Indian	NN	O	O
subcontinent	NN	O	O
.	NN	O	O

The	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
on	NN	O	O
a	NN	O	O
haplotype	NN	O	O
that	NN	O	O
extends	NN	O	O
<	NN	O	O
/	NN	O	O
=	NN	O	O
6	NN	O	O
Mb	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
has	NN	O	O
arisen	NN	O	O
during	NN	O	O
the	NN	O	O
past	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
years	NN	O	O
.	NN	O	O

The	NN	O	O
H63D	NN	O	O
mutation	NN	O	O
is	NN	O	O
older	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
occur	NN	O	O
on	NN	O	O
such	NN	O	O
a	NN	O	O
large	NN	O	O
extended	NN	O	O
haplotype	NN	O	O
,	NN	O	O
the	NN	O	O
haplotype	NN	O	O
in	NN	O	O
this	NN	O	O
case	NN	O	O
extending	NN	O	O
<	NN	O	O
/	NN	O	O
=	NN	O	O
700	NN	O	O
kb	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
finding	NN	O	O
of	NN	O	O
the	NN	O	O
H63D	NN	O	O
and	NN	O	O
C282Y	NN	O	O
mutations	NN	O	O
on	NN	O	O
new	NN	O	O
haplotypes	NN	O	O
.	NN	O	O

In	NN	O	O
Sri	NN	O	O
Lanka	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
H63D	NN	O	O
on	NN	O	O
three	NN	O	O
new	NN	O	O
haplotypes	NN	O	O
and	NN	O	O
have	NN	O	O
found	NN	O	O
C282Y	NN	O	O
on	NN	O	O
one	NN	O	O
new	NN	O	O
haplotype	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
these	NN	O	O
mutations	NN	O	O
have	NN	O	O
arisen	NN	O	O
independently	NN	O	O
on	NN	O	O
this	NN	O	O
island	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
HFE	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
the	NN	O	O
subject	NN	O	O
of	NN	O	O
selection	NN	O	O
pressure	NN	O	O
.	NN	O	O

These	NN	O	O
selection	NN	O	O
pressures	NN	O	O
could	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
infectious	NN	O	B-Disease
diseases	NN	O	I-Disease
,	NN	O	O
environmental	NN	O	O
conditions	NN	O	O
,	NN	O	O
or	NN	O	O
other	NN	O	O
genetic	NN	O	B-Disease
disorders	NN	O	I-Disease
such	NN	O	O
as	NN	O	O
anemia	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
retrospective	NN	O	O
anonymous	NN	O	O
pilot	NN	O	O
study	NN	O	O
in	NN	O	O
screening	NN	O	O
newborns	NN	O	O
for	NN	O	O
HFE	NN	O	O
mutations	NN	O	O
in	NN	O	O
Scandinavian	NN	O	O
populations	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
retrospectively	NN	O	O
analyzed	NN	O	O
837	NN	O	O
random	NN	O	O
anonymized	NN	O	O
dried	NN	O	O
blood	NN	O	O
spot	NN	O	O
(	NN	O	O
DBS	NN	O	O
)	NN	O	O
samples	NN	O	O
from	NN	O	O
neonatal	NN	O	O
screening	NN	O	O
programs	NN	O	O
in	NN	O	O
Scandinavia	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
HFE	NN	O	O
,	NN	O	O
the	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
hemochromatosis	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
C282Y	NN	O	O
allele	NN	O	O
frequencies	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

3	NN	O	O
%	NN	O	O
(	NN	O	O
+	NN	O	O
2	NN	O	O
.	NN	O	O
0	NN	O	O
%	NN	O	O
)	NN	O	O
(	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O
3	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
Greenland	NN	O	O
,	NN	O	O
4	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O

9	NN	O	O
%	NN	O	O
in	NN	O	O
Iceland	NN	O	O
,	NN	O	O
5	NN	O	O
.	NN	O	O

1	NN	O	O
%	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O

3	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
Faeroe	NN	O	O
Islands	NN	O	O
,	NN	O	O
and	NN	O	O
8	NN	O	O
.	NN	O	O

2	NN	O	O
%	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O

7	NN	O	O
%	NN	O	O
in	NN	O	O
Denmark	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
HFE	NN	O	O
mutations	NN	O	O
in	NN	O	O
Denmark	NN	O	O
suggests	NN	O	O
that	NN	O	O
population	NN	O	O
screening	NN	O	O
for	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
could	NN	O	O
be	NN	O	O
highly	NN	O	O
advantageous	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
preventive	NN	O	O
health	NN	O	O
care	NN	O	O
.	NN	O	O

Long	NN	O	O
-	NN	O	O
term	NN	O	O
follow	NN	O	O
-	NN	O	O
up	NN	O	O
evaluation	NN	O	O
of	NN	O	O
C282Y	NN	O	O
homozygotes	NN	O	O
and	NN	O	O
H63D	NN	O	O
/	NN	O	O
C282Y	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
will	NN	O	O
give	NN	O	O
an	NN	O	O
indication	NN	O	O
of	NN	O	O
the	NN	O	O
penetrance	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
alkaptonuria	NN	O	B-Disease
mouse	NN	O	O
model	NN	O	O
.	NN	O	O

Alkaptonuria	NN	O	B-Disease
(	NN	O	O
aku	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
an	NN	O	O
inborn	NN	O	B-Disease
error	NN	O	I-Disease
of	NN	O	I-Disease
metabolism	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
homogentisate	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
-	NN	O	O
dioxygenase	NN	O	O
(	NN	O	O
HGD	NN	O	O
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
a	NN	O	O
mouse	NN	O	O
model	NN	O	O
created	NN	O	O
by	NN	O	O
ethylnitrosourea	NN	O	O
mutagenesis	NN	O	O
but	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
these	NN	O	O
mice	NN	O	O
has	NN	O	O
not	NN	O	O
previously	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
to	NN	O	O
amplify	NN	O	O
the	NN	O	O
Hgd	NN	O	O
cDNA	NN	O	O
from	NN	O	O
Hgd	NN	O	O
(	NN	O	O
aku	NN	O	O
)	NN	O	O
/	NN	O	O
Hgd	NN	O	O
(	NN	O	O
aku	NN	O	O
)	NN	O	O
mice	NN	O	O
.	NN	O	O

Two	NN	O	O
products	NN	O	O
shorter	NN	O	O
than	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
product	NN	O	O
were	NN	O	O
amplified	NN	O	O
.	NN	O	O

Restriction	NN	O	O
mapping	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
were	NN	O	O
then	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
Hgd	NN	O	O
(	NN	O	O
aku	NN	O	O
)	NN	O	O
mouse	NN	O	O
mutation	NN	O	O
,	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
change	NN	O	O
in	NN	O	O
a	NN	O	O
splice	NN	O	O
donor	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
,	NN	O	O
causing	NN	O	O
exon	NN	O	O
skipping	NN	O	O
and	NN	O	O
frame	NN	O	O
-	NN	O	O
shifted	NN	O	O
products	NN	O	O
.	NN	O	O

This	NN	O	O
base	NN	O	O
change	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
create	NN	O	O
a	NN	O	O
non	NN	O	O
-	NN	O	O
radioactive	NN	O	O
genotyping	NN	O	O
assay	NN	O	O
for	NN	O	O
this	NN	O	O
allele	NN	O	O
.	NN	O	O

Non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
enlarged	NN	O	B-Disease
vestibular	NN	O	I-Disease
aqueduct	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
PDS	NN	O	B-Disease
mutations	NN	O	O
.	NN	O	O

Enlarged	NN	O	B-Disease
vestibular	NN	O	I-Disease
aqueduct	NN	O	I-Disease
(	NN	O	O
EVA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
known	NN	O	O
as	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
inner	NN	O	B-Disease
ear	NN	O	I-Disease
abnormality	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
of	NN	O	O
particular	NN	O	O
genetic	NN	O	O
interest	NN	O	O
because	NN	O	O
this	NN	O	O
anomaly	NN	O	O
is	NN	O	O
inherited	NN	O	O
in	NN	O	O
a	NN	O	O
recessive	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
locus	NN	O	O
for	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
sensorineural	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
with	NN	O	O
EVA	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
chromosomal	NN	O	O
region	NN	O	O
,	NN	O	O
7q31	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
locus	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
seven	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
PDS	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
families	NN	O	O
of	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
sensorineural	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
with	NN	O	O
EVA	NN	O	B-Disease
.	NN	O	O

One	NN	O	O
family	NN	O	O
is	NN	O	O
homozygous	NN	O	O
,	NN	O	O
three	NN	O	O
families	NN	O	O
are	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
families	NN	O	O
are	NN	O	O
heterozygous	NN	O	O
but	NN	O	O
with	NN	O	O
no	NN	O	O
other	NN	O	O
mutation	NN	O	O
detected	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
PDS	NN	O	O
cause	NN	O	O
both	NN	O	O
syndromic	NN	O	B-Disease
and	NN	O	I-Disease
non	NN	O	I-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
and	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
of	NN	O	O
29	NN	O	O
unrelated	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
.	NN	O	O

BACKGROUND	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
disease	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
progressive	NN	O	O
neurologic	NN	O	B-Disease
dysfunction	NN	O	I-Disease
,	NN	O	O
occasionally	NN	O	O
associated	NN	O	O
with	NN	O	O
adrenal	NN	O	B-Disease
insufficiency	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
classic	NN	O	O
form	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
usually	NN	O	O
has	NN	O	O
onset	NN	O	O
in	NN	O	O
childhood	NN	O	O
(	NN	O	O
childhood	NN	O	B-Disease
cerebral	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
with	NN	O	O
rapid	NN	O	O
neurologic	NN	O	B-Disease
deterioration	NN	O	I-Disease
leading	NN	O	O
to	NN	O	O
a	NN	O	O
vegetative	NN	O	O
state	NN	O	O
.	NN	O	O

Adult	NN	O	O
-	NN	O	O
onset	NN	O	O
cerebral	NN	O	B-Disease
ALD	NN	O	I-Disease
also	NN	O	O
presents	NN	O	O
with	NN	O	O
rapidly	NN	O	O
progressive	NN	O	O
neurologic	NN	O	B-Disease
dysfunction	NN	O	I-Disease
.	NN	O	O

Milder	NN	O	O
phenotypes	NN	O	O
such	NN	O	O
as	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
and	NN	O	O
Addison	NN	O	B-Disease
disease	NN	O	I-Disease
only	NN	O	O
also	NN	O	O
have	NN	O	O
been	NN	O	O
recognized	NN	O	O
.	NN	O	O

Despite	NN	O	O
discovery	NN	O	O
of	NN	O	O
the	NN	O	O
causative	NN	O	O
gene	NN	O	O
,	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
diverse	NN	O	O
clinical	NN	O	O
presentations	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

OBJECTIVES	NN	O	O
To	NN	O	O
conduct	NN	O	O
mutational	NN	O	O
analyses	NN	O	O
in	NN	O	O
29	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
from	NN	O	O
29	NN	O	O
unrelated	NN	O	O
families	NN	O	O
,	NN	O	O
to	NN	O	O
obtain	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
spectrum	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
study	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O

METHODS	NN	O	O
The	NN	O	O
29	NN	O	O
patients	NN	O	O
comprised	NN	O	O
13	NN	O	O
patients	NN	O	O
with	NN	O	O
childhood	NN	O	B-Disease
cerebral	NN	O	I-Disease
ALD	NN	O	I-Disease
,	NN	O	O
11	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
cerebral	NN	O	B-Disease
ALD	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
5	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
conducted	NN	O	O
detailed	NN	O	O
mutational	NN	O	O
analyses	NN	O	O
of	NN	O	O
29	NN	O	O
unrelated	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
by	NN	O	O
genomic	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
-	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
products	NN	O	O
derived	NN	O	O
from	NN	O	O
total	NN	O	O
RNA	NN	O	O
that	NN	O	O
was	NN	O	O
extracted	NN	O	O
from	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
lymphoblastoid	NN	O	O
cells	NN	O	O
,	NN	O	O
or	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Three	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
cerebral	NN	O	B-Disease
ALD	NN	O	I-Disease
were	NN	O	O
identified	NN	O	O
as	NN	O	O
having	NN	O	O
large	NN	O	O
genomic	NN	O	O
rearrangements	NN	O	O
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
26	NN	O	O
patients	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
having	NN	O	O
21	NN	O	O
independent	NN	O	O
mutations	NN	O	O
,	NN	O	O
including	NN	O	O
12	NN	O	O
novel	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
small	NN	O	O
nucleotide	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Eighteen	NN	O	O
(	NN	O	O
69	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
26	NN	O	O
mutations	NN	O	O
were	NN	O	O
missense	NN	O	O
mutations	NN	O	O
.	NN	O	O

Most	NN	O	O
missense	NN	O	O
mutations	NN	O	O
involved	NN	O	O
amino	NN	O	O
acids	NN	O	O
conserved	NN	O	O
in	NN	O	O
homologous	NN	O	O
gene	NN	O	O
products	NN	O	O
,	NN	O	O
including	NN	O	O
PMP70	NN	O	O
,	NN	O	O
mALDRP	NN	O	O
,	NN	O	O
and	NN	O	O
Pxa1p	NN	O	O
.	NN	O	O

The	NN	O	O
AG	NN	O	O
dinucleotide	NN	O	O
deletion	NN	O	O
at	NN	O	O
position	NN	O	O
1081	NN	O	O
-	NN	O	O
1082	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
in	NN	O	O
white	NN	O	O
patients	NN	O	O
(	NN	O	O
12	NN	O	O
%	NN	O	O
-	NN	O	O
17	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
(	NN	O	O
12	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

All	NN	O	O
phenotypes	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
protein	NN	O	O
truncation	NN	O	O
or	NN	O	O
subtle	NN	O	O
amino	NN	O	O
acid	NN	O	O
changes	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
differences	NN	O	O
in	NN	O	O
phenotypic	NN	O	O
expressions	NN	O	O
between	NN	O	O
missense	NN	O	O
mutations	NN	O	O
involving	NN	O	O
conserved	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
those	NN	O	O
involving	NN	O	O
nonconserved	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
There	NN	O	O
are	NN	O	O
no	NN	O	O
obvious	NN	O	O
correlations	NN	O	O
between	NN	O	O
the	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
their	NN	O	O
genotypes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
other	NN	O	O
genetic	NN	O	O
or	NN	O	O
environmental	NN	O	O
factors	NN	O	O
modify	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
expressions	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
common	NN	O	O
human	NN	O	O
skin	NN	O	B-Disease
tumour	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
activating	NN	O	O
mutations	NN	O	O
in	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
.	NN	O	O

WNT	NN	O	O
signalling	NN	O	O
orchestrates	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
developmental	NN	O	O
programs	NN	O	O
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
this	NN	O	O
stimulus	NN	O	O
,	NN	O	O
cytoplasmic	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
(	NN	O	O
encoded	NN	O	O
by	NN	O	O
CTNNB1	NN	O	O
)	NN	O	O
is	NN	O	O
stabilized	NN	O	O
,	NN	O	O
enabling	NN	O	O
downstream	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
LEF	NN	O	O
/	NN	O	O
TCF	NN	O	O
family	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	O
genes	NN	O	O
for	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
/	NN	O	O
TCF	NN	O	O
encodes	NN	O	O
c	NN	O	O
-	NN	O	O
MYC	NN	O	O
,	NN	O	O
explaining	NN	O	O
why	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
WNT	NN	O	O
pathway	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
the	NN	O	O
colon	NN	O	O
.	NN	O	O

Most	NN	O	O
colon	NN	O	B-Disease
cancers	NN	O	I-Disease
arise	NN	O	O
from	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	O
required	NN	O	O
for	NN	O	O
ubiquitin	NN	O	O
-	NN	O	O
mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
,	NN	O	O
but	NN	O	O
a	NN	O	O
small	NN	O	O
percentage	NN	O	O
of	NN	O	O
colon	NN	O	B-Disease
and	NN	O	I-Disease
some	NN	O	I-Disease
other	NN	O	I-Disease
cancers	NN	O	I-Disease
harbour	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
-	NN	O	O
stabilizing	NN	O	O
mutations	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
discovered	NN	O	O
that	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
an	NN	O	O
activated	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
are	NN	O	O
predisposed	NN	O	O
to	NN	O	O
developing	NN	O	O
skin	NN	O	B-Disease
tumours	NN	O	I-Disease
resembling	NN	O	O
pilomatricomas	NN	O	B-Disease
.	NN	O	O

Given	NN	O	O
that	NN	O	O
the	NN	O	O
skin	NN	O	O
of	NN	O	O
these	NN	O	O
adult	NN	O	O
mice	NN	O	O
also	NN	O	O
exhibits	NN	O	O
signs	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
hair	NN	O	O
-	NN	O	O
follicle	NN	O	O
morphogenesis	NN	O	O
,	NN	O	O
we	NN	O	O
wondered	NN	O	O
whether	NN	O	O
human	NN	O	O
pilomatricomas	NN	O	B-Disease
might	NN	O	O
originate	NN	O	O
from	NN	O	O
hair	NN	O	O
matrix	NN	O	O
cells	NN	O	O
and	NN	O	O
whether	NN	O	O
they	NN	O	O
might	NN	O	O
possess	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
-	NN	O	O
stabilizing	NN	O	O
mutations	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
explore	NN	O	O
the	NN	O	O
cell	NN	O	O
origin	NN	O	O
and	NN	O	O
aetiology	NN	O	O
of	NN	O	O
this	NN	O	O
common	NN	O	O
human	NN	O	O
skin	NN	O	B-Disease
tumour	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
found	NN	O	O
nuclear	NN	O	O
LEF	NN	O	O
-	NN	O	O
1	NN	O	O
in	NN	O	O
the	NN	O	O
dividing	NN	O	O
tumour	NN	O	B-Disease
cells	NN	O	O
,	NN	O	O
providing	NN	O	O
biochemical	NN	O	O
evidence	NN	O	O
that	NN	O	O
pilomatricomas	NN	O	B-Disease
are	NN	O	O
derived	NN	O	O
from	NN	O	O
hair	NN	O	O
matrix	NN	O	O
cells	NN	O	O
.	NN	O	O

At	NN	O	O
least	NN	O	O
75	NN	O	O
%	NN	O	O
of	NN	O	O
these	NN	O	O
tumours	NN	O	B-Disease
possess	NN	O	O
mutations	NN	O	O
affecting	NN	O	O
the	NN	O	O
amino	NN	O	O
-	NN	O	O
terminal	NN	O	O
segment	NN	O	O
,	NN	O	O
normally	NN	O	O
involved	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
-	NN	O	O
dependent	NN	O	O
,	NN	O	O
ubiquitin	NN	O	O
-	NN	O	O
mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

This	NN	O	O
percentage	NN	O	O
of	NN	O	O
CTNNB1	NN	O	O
mutations	NN	O	O
is	NN	O	O
greater	NN	O	O
than	NN	O	O
in	NN	O	O
all	NN	O	O
other	NN	O	O
human	NN	O	O
tumours	NN	O	B-Disease
examined	NN	O	O
thus	NN	O	O
far	NN	O	O
,	NN	O	O
and	NN	O	O
directly	NN	O	O
implicates	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
/	NN	O	O
LEF	NN	O	O
misregulation	NN	O	O
as	NN	O	O
the	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
hair	NN	O	O
matrix	NN	O	O
cell	NN	O	O
tumorigenesis	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
chloride	NN	O	O
-	NN	O	O
iodide	NN	O	O
transport	NN	O	O
protein	NN	O	O
.	NN	O	O

Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
syndromic	NN	O	B-Disease
deafness	NN	O	I-Disease
and	NN	O	O
characterized	NN	O	O
by	NN	O	O
congenital	NN	O	O
sensorineural	NN	O	B-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
and	NN	O	O
goitre	NN	O	B-Disease
.	NN	O	O

This	NN	O	O
disorder	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
7	NN	O	O
and	NN	O	O
the	NN	O	O
gene	NN	O	O
causing	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PDS	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
subsequently	NN	O	O
identified	NN	O	O
by	NN	O	O
positional	NN	O	O
cloning	NN	O	O
.	NN	O	O

PDS	NN	O	O
encodes	NN	O	O
a	NN	O	O
putative	NN	O	O
transmembrane	NN	O	O
protein	NN	O	O
designated	NN	O	O
pendrin	NN	O	O
.	NN	O	O

Pendrin	NN	O	O
is	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
sulfate	NN	O	O
transport	NN	O	O
proteins	NN	O	O
that	NN	O	O
includes	NN	O	O
the	NN	O	O
rat	NN	O	O
sulfate	NN	O	O
-	NN	O	O
anion	NN	O	O
transporter	NN	O	O
(	NN	O	O
encoded	NN	O	O
by	NN	O	O
Sat	NN	O	O
-	NN	O	O
1	NN	O	O
;	NN	O	O
29	NN	O	O
%	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
identity	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
sulfate	NN	O	O
transporter	NN	O	O
(	NN	O	O
encoded	NN	O	O
by	NN	O	O
DTD	NN	O	O
;	NN	O	O
32	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
human	NN	O	O
sulfate	NN	O	O
transporter	NN	O	O
downregulated	NN	O	O
in	NN	O	O
adenoma	NN	O	B-Disease
(	NN	O	O
encoded	NN	O	O
by	NN	O	O
DRA	NN	O	O
;	NN	O	O
45	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
this	NN	O	O
homology	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
slightly	NN	O	O
modified	NN	O	O
sulfate	NN	O	O
-	NN	O	O
transporter	NN	O	O
signature	NN	O	O
sequence	NN	O	O
comprising	NN	O	O
its	NN	O	O
putative	NN	O	O
second	NN	O	O
transmembrane	NN	O	O
domain	NN	O	O
,	NN	O	O
pendrin	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
sulfate	NN	O	O
transporter	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
detect	NN	O	O
evidence	NN	O	O
of	NN	O	O
sulfate	NN	O	O
transport	NN	O	O
following	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
pendrin	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
laevis	NN	O	O
oocytes	NN	O	O
by	NN	O	O
microinjection	NN	O	O
of	NN	O	O
PDS	NN	O	O
cRNA	NN	O	O
or	NN	O	O
in	NN	O	O
Sf9	NN	O	O
cells	NN	O	O
following	NN	O	O
infection	NN	O	O
with	NN	O	O
PDS	NN	O	O
-	NN	O	O
recombinant	NN	O	O
baculovirus	NN	O	O
.	NN	O	O

The	NN	O	O
rates	NN	O	O
of	NN	O	O
transport	NN	O	O
for	NN	O	O
iodide	NN	O	O
and	NN	O	O
chloride	NN	O	O
were	NN	O	O
significantly	NN	O	O
increased	NN	O	O
following	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
pendrin	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
systems	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
pendrin	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transporter	NN	O	O
of	NN	O	O
chloride	NN	O	O
and	NN	O	O
iodide	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
sulfate	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
thyroid	NN	O	O
physiology	NN	O	O
and	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

HFE	NN	O	O
mutations	NN	O	O
analysis	NN	O	O
in	NN	O	O
711	NN	O	O
hemochromatosis	NN	O	B-Disease
probands	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
S65C	NN	O	O
implication	NN	O	O
in	NN	O	O
mild	NN	O	O
form	NN	O	O
of	NN	O	O
hemochromatosis	NN	O	B-Disease
.	NN	O	O

Hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
(	NN	O	O
HH	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	O
iron	NN	O	O
metabolism	NN	O	O
.	NN	O	O

The	NN	O	O
HFE	NN	O	O
candidate	NN	O	O
gene	NN	O	O
encoding	NN	O	O
an	NN	O	O
HLA	NN	O	O
class	NN	O	O
I	NN	O	O
-	NN	O	O
like	NN	O	O
protein	NN	O	O
involved	NN	O	O
in	NN	O	O
HH	NN	O	B-Disease
was	NN	O	O
identified	NN	O	O
in	NN	O	O
1996	NN	O	O
.	NN	O	O

Two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
C282Y	NN	O	O
,	NN	O	O
accounting	NN	O	O
for	NN	O	O
80	NN	O	O
%	NN	O	O
to	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
HH	NN	O	B-Disease
chromosomes	NN	O	O
,	NN	O	O
and	NN	O	O
H63D	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
milder	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
representing	NN	O	O
40	NN	O	O
%	NN	O	O
to	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
C282Y	NN	O	O
HH	NN	O	B-Disease
chromosomes	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
on	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
C282Y	NN	O	O
,	NN	O	O
H63D	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
193A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
substitution	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
S65C	NN	O	O
missense	NN	O	O
substitution	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
series	NN	O	O
of	NN	O	O
probands	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
confirm	NN	O	O
that	NN	O	O
the	NN	O	O
C282Y	NN	O	O
substitution	NN	O	O
was	NN	O	O
the	NN	O	O
main	NN	O	O
mutation	NN	O	O
involved	NN	O	O
in	NN	O	O
hemochromatosis	NN	O	B-Disease
,	NN	O	O
accounting	NN	O	O
for	NN	O	O
85	NN	O	O
%	NN	O	O
of	NN	O	O
carrier	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
H63D	NN	O	O
substitution	NN	O	O
represented	NN	O	O
39	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
HH	NN	O	B-Disease
chromosomes	NN	O	O
that	NN	O	O
did	NN	O	O
not	NN	O	O
carry	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
our	NN	O	O
screening	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
S65C	NN	O	O
substitution	NN	O	O
was	NN	O	O
significantly	NN	O	O
enriched	NN	O	O
in	NN	O	O
probands	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
chromosome	NN	O	O
without	NN	O	O
an	NN	O	O
assigned	NN	O	O
mutation	NN	O	O
.	NN	O	O

This	NN	O	O
substitution	NN	O	O
accounted	NN	O	O
for	NN	O	O
7	NN	O	O
.	NN	O	O

8	NN	O	O
%	NN	O	O
of	NN	O	O
HH	NN	O	B-Disease
chromosomes	NN	O	O
that	NN	O	O
were	NN	O	O
neither	NN	O	O
C282Y	NN	O	O
nor	NN	O	O
H63D	NN	O	O
.	NN	O	O

This	NN	O	O
enrichment	NN	O	O
of	NN	O	O
S65C	NN	O	O
among	NN	O	O
HH	NN	O	B-Disease
chromosomes	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
S65C	NN	O	O
substitution	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
mild	NN	O	O
form	NN	O	O
of	NN	O	O
hemochromatosis	NN	O	B-Disease
.	NN	O	O

Germline	NN	O	O
BRCA1	NN	O	O
alterations	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
series	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
cases	NN	O	O
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
provide	NN	O	O
more	NN	O	O
accurate	NN	O	O
frequency	NN	O	O
estimates	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
gene	NN	O	O
1	NN	O	O
(	NN	O	O
BRCA1	NN	O	O
)	NN	O	O
germline	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
population	NN	O	O
.	NN	O	O

To	NN	O	O
achieve	NN	O	O
this	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
alterations	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
series	NN	O	O
of	NN	O	O
consecutive	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
cases	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
study	NN	O	O
reporting	NN	O	O
BRCA1	NN	O	O
alterations	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
screen	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
.	NN	O	O

One	NN	O	O
hundred	NN	O	O
and	NN	O	O
seven	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
cases	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
alterations	NN	O	O
using	NN	O	O
the	NN	O	O
RNase	NN	O	O
mismatch	NN	O	O
cleavage	NN	O	O
assay	NN	O	O
followed	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Two	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
,	NN	O	O
962del4	NN	O	O
and	NN	O	O
3600del11	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Both	NN	O	O
patients	NN	O	O
had	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Several	NN	O	O
novel	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
previously	NN	O	O
reported	NN	O	O
uncharacterized	NN	O	O
variants	NN	O	O
were	NN	O	O
also	NN	O	O
identified	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
distribution	NN	O	O
of	NN	O	O
common	NN	O	O
polymorphisms	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
the	NN	O	O
91	NN	O	O
Caucasian	NN	O	O
cancer	NN	O	B-Disease
cases	NN	O	O
in	NN	O	O
this	NN	O	O
series	NN	O	O
and	NN	O	O
24	NN	O	O
sister	NN	O	O
controls	NN	O	O
using	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
amplification	NN	O	O
.	NN	O	O

The	NN	O	O
rare	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
Q356R	NN	O	O
polymorphism	NN	O	O
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
polymorphism	NN	O	O
may	NN	O	O
influence	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
risk	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
some	NN	O	O
uncharacterized	NN	O	O
variants	NN	O	O
and	NN	O	O
rare	NN	O	O
forms	NN	O	O
of	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
determining	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
risk	NN	O	O
,	NN	O	O
and	NN	O	O
highlight	NN	O	O
the	NN	O	O
necessity	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
missense	NN	O	O
alterations	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	O
-	NN	O	O
related	NN	O	O
protein	NN	O	O
can	NN	O	O
compensate	NN	O	O
functionally	NN	O	O
for	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
protein	NN	O	I-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
therapy	NN	O	O
.	NN	O	O

Inherited	NN	O	B-Disease
defects	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
peroxisomal	NN	O	O
ATP	NN	O	O
-	NN	O	O
binding	NN	O	O
cassette	NN	O	O
(	NN	O	O
ABC	NN	O	O
)	NN	O	O
transporter	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
protein	NN	O	O
(	NN	O	O
ALDP	NN	O	O
)	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
lethal	NN	O	O
peroxisomal	NN	O	B-Disease
disorder	NN	O	I-Disease
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
for	NN	O	O
which	NN	O	O
no	NN	O	O
efficient	NN	O	O
treatment	NN	O	O
has	NN	O	O
been	NN	O	O
established	NN	O	O
so	NN	O	O
far	NN	O	O
.	NN	O	O

Three	NN	O	O
other	NN	O	O
peroxisomal	NN	O	O
ABC	NN	O	O
transporters	NN	O	O
currently	NN	O	O
are	NN	O	O
known	NN	O	O
adrenoleukodystrophy	NN	O	O
-	NN	O	O
related	NN	O	O
protein	NN	O	O
(	NN	O	O
ALDRP	NN	O	O
)	NN	O	O
,	NN	O	O
70	NN	O	O
kDa	NN	O	O
peroxisomal	NN	O	O
membrane	NN	O	O
protein	NN	O	O
(	NN	O	O
PMP70	NN	O	O
)	NN	O	O
and	NN	O	O
PMP70	NN	O	O
-	NN	O	O
related	NN	O	O
protein	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
transient	NN	O	O
and	NN	O	O
stable	NN	O	O
overexpression	NN	O	O
of	NN	O	O
human	NN	O	O
cDNAs	NN	O	O
encoding	NN	O	O
ALDP	NN	O	O
and	NN	O	O
its	NN	O	O
closest	NN	O	O
relative	NN	O	O
ALDRP	NN	O	O
,	NN	O	O
we	NN	O	O
could	NN	O	O
restore	NN	O	O
the	NN	O	O
impaired	NN	O	O
peroxisomal	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
in	NN	O	O
fibroblasts	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

The	NN	O	O
pathognomonic	NN	O	O
accumulation	NN	O	O
of	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
could	NN	O	O
also	NN	O	O
be	NN	O	O
prevented	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
ALDRP	NN	O	O
in	NN	O	O
immortalized	NN	O	O
X	NN	O	O
-	NN	O	O
ALD	NN	O	O
cells	NN	O	O
.	NN	O	O

Immunofluorescence	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
replacement	NN	O	O
of	NN	O	O
ALDP	NN	O	O
by	NN	O	O
ALDRP	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
mutated	NN	O	O
ALDP	NN	O	O
itself	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
restore	NN	O	O
the	NN	O	O
peroxisomal	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
of	NN	O	O
ALDP	NN	O	O
-	NN	O	O
deficient	NN	O	O
mice	NN	O	O
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
ALDRP	NN	O	O
and	NN	O	O
PMP70	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
a	NN	O	O
dietary	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
peroxisome	NN	O	O
proliferator	NN	O	O
fenofibrate	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
correction	NN	O	O
of	NN	O	O
the	NN	O	O
biochemical	NN	O	O
defect	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
could	NN	O	O
be	NN	O	O
possible	NN	O	O
by	NN	O	O
drug	NN	O	O
-	NN	O	O
induced	NN	O	O
overexpression	NN	O	O
or	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
ALDRP	NN	O	O
.	NN	O	O
.	NN	O	O

Centrosome	NN	O	O
amplification	NN	O	O
and	NN	O	O
a	NN	O	O
defective	NN	O	O
G2	NN	O	O
-	NN	O	O
M	NN	O	O
cell	NN	O	O
cycle	NN	O	O
checkpoint	NN	O	O
induce	NN	O	O
genetic	NN	O	O
instability	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
exon	NN	O	O
11	NN	O	O
isoform	NN	O	O
-	NN	O	O
deficient	NN	O	O
cells	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
Brca1	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
predispose	NN	O	O
women	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
study	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
related	NN	O	O
tumorigenesis	NN	O	O
,	NN	O	O
we	NN	O	O
derived	NN	O	O
mouse	NN	O	O
embryonic	NN	O	O
fibroblast	NN	O	O
cells	NN	O	O
carrying	NN	O	O
a	NN	O	O
targeted	NN	O	O
deletion	NN	O	O
of	NN	O	O
exon	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
Brca1	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
mutant	NN	O	O
cells	NN	O	O
maintain	NN	O	O
an	NN	O	O
intact	NN	O	O
G1	NN	O	O
-	NN	O	O
S	NN	O	O
cell	NN	O	O
cycle	NN	O	O
checkpoint	NN	O	O
and	NN	O	O
proliferate	NN	O	O
poorly	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
defective	NN	O	O
G2	NN	O	O
-	NN	O	O
M	NN	O	O
checkpoint	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
extensive	NN	O	O
chromosomal	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

Mutant	NN	O	O
fibroblasts	NN	O	O
contain	NN	O	O
multiple	NN	O	O
,	NN	O	O
functional	NN	O	O
centrosomes	NN	O	O
,	NN	O	O
which	NN	O	O
lead	NN	O	O
to	NN	O	O
unequal	NN	O	O
chromosome	NN	O	O
segregation	NN	O	O
,	NN	O	O
abnormal	NN	O	O
nuclear	NN	O	O
division	NN	O	O
,	NN	O	O
and	NN	O	O
aneuploidy	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
data	NN	O	O
uncover	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
maintaining	NN	O	O
genetic	NN	O	O
stability	NN	O	O
through	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
centrosome	NN	O	O
duplication	NN	O	O
and	NN	O	O
the	NN	O	O
G2	NN	O	O
-	NN	O	O
M	NN	O	O
checkpoint	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O
.	NN	O	O

Defective	NN	O	O
CD95	NN	O	O
/	NN	O	O
APO	NN	O	O
-	NN	O	O
1	NN	O	O
/	NN	O	O
Fas	NN	O	O
signal	NN	O	O
complex	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	I-Disease
type	NN	O	I-Disease
Ia	NN	O	I-Disease
.	NN	O	O

Heterozygous	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
CD95	NN	O	O
(	NN	O	O
APO	NN	O	O
-	NN	O	O
1	NN	O	O
/	NN	O	O
Fas	NN	O	O
)	NN	O	O
receptor	NN	O	O
occur	NN	O	O
in	NN	O	O
most	NN	O	O
individuals	NN	O	O
with	NN	O	O
autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ALPS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
dominantly	NN	O	O
interfere	NN	O	O
with	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
an	NN	O	O
unknown	NN	O	O
mechanism	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
local	NN	O	O
or	NN	O	O
global	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
death	NN	O	O
domain	NN	O	O
from	NN	O	O
nine	NN	O	O
independent	NN	O	O
ALPS	NN	O	B-Disease
CD95	NN	O	O
death	NN	O	O
-	NN	O	O
domain	NN	O	O
mutations	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
FADD	NN	O	O
/	NN	O	O
MORT1	NN	O	O
signaling	NN	O	O
protein	NN	O	O
.	NN	O	O

Despite	NN	O	O
heterozygosity	NN	O	O
for	NN	O	O
the	NN	O	O
abnormal	NN	O	O
allele	NN	O	O
,	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
ALPS	NN	O	B-Disease
patients	NN	O	O
showed	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
FADD	NN	O	O
association	NN	O	O
and	NN	O	O
a	NN	O	O
loss	NN	O	O
of	NN	O	O
caspase	NN	O	O
recruitment	NN	O	O
and	NN	O	O
activation	NN	O	O
after	NN	O	O
CD95	NN	O	O
crosslinking	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
intracytoplasmic	NN	O	O
CD95	NN	O	O
mutations	NN	O	O
in	NN	O	O
ALPS	NN	O	B-Disease
impair	NN	O	O
apoptosis	NN	O	O
chiefly	NN	O	O
by	NN	O	O
disrupting	NN	O	O
death	NN	O	O
-	NN	O	O
domain	NN	O	O
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
signaling	NN	O	O
protein	NN	O	O
FADD	NN	O	O
/	NN	O	O
MORT1	NN	O	O
.	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
alkaptonuria	NN	O	B-Disease
(	NN	O	O
AKU	NN	O	B-Disease
)	NN	O	O
mutations	NN	O	O
and	NN	O	O
polymorphisms	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
CCC	NN	O	O
sequence	NN	O	O
motif	NN	O	O
is	NN	O	O
a	NN	O	O
mutational	NN	O	O
hot	NN	O	O
spot	NN	O	O
in	NN	O	O
the	NN	O	O
homogentisate	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
dioxygenase	NN	O	O
gene	NN	O	O
(	NN	O	O
HGO	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
showed	NN	O	O
that	NN	O	O
alkaptonuria	NN	O	B-Disease
(	NN	O	O
AKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
loss	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
function	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
homogentisate	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
dioxygenase	NN	O	O
gene	NN	O	O
(	NN	O	O
HGO	NN	O	O
)	NN	O	O
.	NN	O	O

Herein	NN	O	O
we	NN	O	O
describe	NN	O	O
haplotype	NN	O	O
and	NN	O	O
mutational	NN	O	O
analyses	NN	O	O
of	NN	O	O
HGO	NN	O	O
in	NN	O	O
seven	NN	O	O
new	NN	O	O
AKU	NN	O	B-Disease
pedigrees	NN	O	O
.	NN	O	O

These	NN	O	O
analyses	NN	O	O
identified	NN	O	O
two	NN	O	O
novel	NN	O	O
single	NN	O	O
-	NN	O	O
nucleotide	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
INV4	NN	O	O
+	NN	O	O
31A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
and	NN	O	O
INV11	NN	O	O
+	NN	O	O
18A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
and	NN	O	O
six	NN	O	O
novel	NN	O	O
AKU	NN	O	B-Disease
mutations	NN	O	O
(	NN	O	O
INV1	NN	O	O
-	NN	O	O
1G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
W60G	NN	O	O
,	NN	O	O
Y62C	NN	O	O
,	NN	O	O
A122D	NN	O	O
,	NN	O	O
P230T	NN	O	O
,	NN	O	O
and	NN	O	O
D291E	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
further	NN	O	O
illustrates	NN	O	O
the	NN	O	O
remarkable	NN	O	O
allelic	NN	O	O
heterogeneity	NN	O	O
found	NN	O	O
in	NN	O	O
AKU	NN	O	B-Disease
.	NN	O	O

Reexamination	NN	O	O
of	NN	O	O
all	NN	O	O
29	NN	O	O
mutations	NN	O	O
and	NN	O	O
polymorphisms	NN	O	O
thus	NN	O	O
far	NN	O	O
described	NN	O	O
in	NN	O	O
HGO	NN	O	O
shows	NN	O	O
that	NN	O	O
these	NN	O	O
nucleotide	NN	O	O
changes	NN	O	O
are	NN	O	O
not	NN	O	O
randomly	NN	O	O
distributed	NN	O	O
;	NN	O	O
the	NN	O	O
CCC	NN	O	O
sequence	NN	O	O
motif	NN	O	O
and	NN	O	O
its	NN	O	O
inverted	NN	O	O
complement	NN	O	O
,	NN	O	O
GGG	NN	O	O
,	NN	O	O
are	NN	O	O
preferentially	NN	O	O
mutated	NN	O	O
.	NN	O	O

These	NN	O	O
analyses	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
substitutions	NN	O	O
in	NN	O	O
HGO	NN	O	O
do	NN	O	O
not	NN	O	O
involve	NN	O	O
CpG	NN	O	O
dinucleotides	NN	O	O
,	NN	O	O
which	NN	O	O
illustrates	NN	O	O
important	NN	O	O
differences	NN	O	O
between	NN	O	O
HGO	NN	O	O
and	NN	O	O
other	NN	O	O
genes	NN	O	O
for	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
mutation	NN	O	O
at	NN	O	O
specific	NN	O	O
short	NN	O	O
-	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
CCC	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
comprise	NN	O	O
a	NN	O	O
significant	NN	O	O
proportion	NN	O	O
(	NN	O	O
34	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
all	NN	O	O
mutated	NN	O	O
bases	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
HGO	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
CCC	NN	O	O
triplet	NN	O	O
is	NN	O	O
a	NN	O	O
mutational	NN	O	O
hot	NN	O	O
spot	NN	O	O
in	NN	O	O
HGO	NN	O	O
.	NN	O	O

Fabry	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
identification	NN	O	O
of	NN	O	O
novel	NN	O	O
alpha	NN	O	O
-	NN	O	O
galactosidase	NN	O	O
A	NN	O	O
mutations	NN	O	O
and	NN	O	O
molecular	NN	O	O
carrier	NN	O	O
detection	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
fluorescent	NN	O	O
chemical	NN	O	O
cleavage	NN	O	O
of	NN	O	O
mismatches	NN	O	O
.	NN	O	O

Fabry	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
FD	NN	O	B-Disease
)	NN	O	O
(	NN	O	O
angiokeratoma	NN	O	B-Disease
corporis	NN	O	I-Disease
diffusum	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
inborn	NN	O	I-Disease
error	NN	O	I-Disease
of	NN	O	I-Disease
glycosphingolipid	NN	O	I-Disease
metabolism	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
alpha	NN	O	O
-	NN	O	O
galactosidase	NN	O	O
A	NN	O	O
gene	NN	O	O
(	NN	O	O
GLA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
enzymatic	NN	O	O
defect	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
systemic	NN	O	O
accumulation	NN	O	O
of	NN	O	O
neutral	NN	O	O
glycosphingolipids	NN	O	O
with	NN	O	O
terminal	NN	O	O
alpha	NN	O	O
-	NN	O	O
galactosyl	NN	O	O
moieties	NN	O	O
.	NN	O	O

Clinically	NN	O	O
,	NN	O	O
affected	NN	O	O
hemizygous	NN	O	O
males	NN	O	O
have	NN	O	O
angiokeratoma	NN	O	B-Disease
,	NN	O	O
severe	NN	O	B-Disease
acroparesthesia	NN	O	I-Disease
,	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
vasculopathy	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
heart	NN	O	I-Disease
and	NN	O	I-Disease
brain	NN	O	I-Disease
.	NN	O	O

While	NN	O	O
demonstration	NN	O	O
of	NN	O	O
alpha	NN	O	B-Disease
-	NN	O	I-Disease
galactosidase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
leukocytes	NN	O	O
is	NN	O	O
diagnostic	NN	O	O
in	NN	O	O
affected	NN	O	O
males	NN	O	O
,	NN	O	O
enzymatic	NN	O	O
detection	NN	O	O
of	NN	O	O
female	NN	O	O
carriers	NN	O	O
is	NN	O	O
often	NN	O	O
inconclusive	NN	O	O
,	NN	O	O
due	NN	O	O
to	NN	O	O
random	NN	O	O
X	NN	O	O
-	NN	O	O
chromosomal	NN	O	O
inactivation	NN	O	O
,	NN	O	O
underlining	NN	O	O
the	NN	O	O
need	NN	O	O
of	NN	O	O
molecular	NN	O	O
investigations	NN	O	O
for	NN	O	O
accurate	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
.	NN	O	O

By	NN	O	O
use	NN	O	O
of	NN	O	O
chemical	NN	O	O
cleavage	NN	O	O
of	NN	O	O
mismatches	NN	O	O
adapted	NN	O	O
to	NN	O	O
fluorescence	NN	O	O
-	NN	O	O
based	NN	O	O
detection	NN	O	O
systems	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
mutations	NN	O	O
underlying	NN	O	O
alpha	NN	O	B-Disease
-	NN	O	I-Disease
Gal	NN	O	I-Disease
A	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
16	NN	O	O
individuals	NN	O	O
from	NN	O	O
six	NN	O	O
unrelated	NN	O	O
families	NN	O	O
with	NN	O	O
FD	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
mutational	NN	O	O
spectrum	NN	O	O
included	NN	O	O
five	NN	O	O
missense	NN	O	O
mutations	NN	O	O
(	NN	O	O
C202W	NN	O	O
,	NN	O	O
C223G	NN	O	O
,	NN	O	O
N224D	NN	O	O
,	NN	O	O
R301Q	NN	O	O
,	NN	O	O
and	NN	O	O
Q327K	NN	O	O
)	NN	O	O
and	NN	O	O
one	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
[	NN	O	O
IVS3	NN	O	O
G	NN	O	O
(	NN	O	O
-	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
]	NN	O	O
.	NN	O	O

Studies	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
latter	NN	O	O
led	NN	O	O
to	NN	O	O
altered	NN	O	O
pre	NN	O	O
-	NN	O	O
mRNA	NN	O	O
splicing	NN	O	O
with	NN	O	O
consequent	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
mRNA	NN	O	O
translational	NN	O	O
reading	NN	O	O
frame	NN	O	O
and	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
premature	NN	O	O
termination	NN	O	O
codon	NN	O	O
of	NN	O	O
translation	NN	O	O
.	NN	O	O

By	NN	O	O
use	NN	O	O
of	NN	O	O
this	NN	O	O
strategy	NN	O	O
,	NN	O	O
carrier	NN	O	O
status	NN	O	O
was	NN	O	O
accurately	NN	O	O
assessed	NN	O	O
in	NN	O	O
all	NN	O	O
seven	NN	O	O
at	NN	O	O
-	NN	O	O
risk	NN	O	O
females	NN	O	O
tested	NN	O	O
,	NN	O	O
whereas	NN	O	O
enzymatic	NN	O	O
dosages	NN	O	O
failed	NN	O	O
to	NN	O	O
diagnose	NN	O	O
or	NN	O	O
exclude	NN	O	O
heterozygosity	NN	O	O
.	NN	O	O
.	NN	O	O

Prenatal	NN	O	O
diagnosis	NN	O	O
by	NN	O	O
FISH	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
duplication	NN	O	O
of	NN	O	O
PLP	NN	O	O
gene	NN	O	O
.	NN	O	O

A	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MIM	NN	O	O
312080	NN	O	O
)	NN	O	O
was	NN	O	O
made	NN	O	O
in	NN	O	O
a	NN	O	O
young	NN	O	O
boy	NN	O	O
.	NN	O	O

No	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
had	NN	O	O
been	NN	O	O
found	NN	O	O
.	NN	O	O

The	NN	O	O
boys	NN	O	O
maternal	NN	O	O
aunt	NN	O	O
came	NN	O	O
for	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
when	NN	O	O
16	NN	O	O
+	NN	O	O
weeks	NN	O	O
pregnant	NN	O	O
and	NN	O	O
carrying	NN	O	O
a	NN	O	O
male	NN	O	O
fetus	NN	O	O
.	NN	O	O

Samples	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
duplication	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
,	NN	O	O
by	NN	O	O
interphase	NN	O	O
FISH	NN	O	O
,	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
preparations	NN	O	O
from	NN	O	O
the	NN	O	O
proband	NN	O	O
,	NN	O	O
his	NN	O	O
aunt	NN	O	O
and	NN	O	O
an	NN	O	O
amniotic	NN	O	O
fluid	NN	O	O
cell	NN	O	O
preparation	NN	O	O
from	NN	O	O
the	NN	O	O
fetus	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
the	NN	O	O
duplication	NN	O	O
,	NN	O	O
thus	NN	O	O
confirming	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
neither	NN	O	O
the	NN	O	O
aunt	NN	O	O
nor	NN	O	O
the	NN	O	O
fetus	NN	O	O
carried	NN	O	O
a	NN	O	O
duplication	NN	O	O
.	NN	O	O
.	NN	O	O

Dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
APC1309	NN	O	O
mutation	NN	O	O
:	NN	O	O
a	NN	O	O
possible	NN	O	O
explanation	NN	O	O
for	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
.	NN	O	O

Inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
product	NN	O	O
initiates	NN	O	O
colorectal	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
APC	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
carry	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
develop	NN	O	O
multiple	NN	O	O
colorectal	NN	O	B-Disease
adenomas	NN	O	I-Disease
and	NN	O	O
subsequent	NN	O	O
carcinomas	NN	O	B-Disease
early	NN	O	O
in	NN	O	O
life	NN	O	O
.	NN	O	O

The	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
inherited	NN	O	O
APC	NN	O	B-Disease
mutation	NN	O	O
(	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
)	NN	O	O
.	NN	O	O

Together	NN	O	O
with	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
both	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
and	NN	O	O
sporadic	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
cluster	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
this	NN	O	O
points	NN	O	O
to	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
certain	NN	O	O
APC	NN	O	B-Disease
mutants	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
function	NN	O	O
was	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
enhanced	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
-	NN	O	O
/	NN	O	O
Tcf	NN	O	O
-	NN	O	O
mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
colon	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
experimental	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
products	NN	O	O
associated	NN	O	O
with	NN	O	O
severe	NN	O	O
polyposis	NN	O	B-Disease
.	NN	O	O

Wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	B-Disease
activity	NN	O	O
in	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
-	NN	O	O
/	NN	O	O
Tcf	NN	O	O
-	NN	O	O
mediated	NN	O	O
transcription	NN	O	O
was	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
mutant	NN	O	O
APC	NN	O	B-Disease
that	NN	O	O
is	NN	O	O
truncated	NN	O	O
at	NN	O	O
codon	NN	O	O
1309	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mutant	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
products	NN	O	O
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
attenuated	NN	O	B-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
codon	NN	O	O
386	NN	O	O
or	NN	O	O
1465	NN	O	O
)	NN	O	O
interfered	NN	O	O
only	NN	O	O
weakly	NN	O	O
with	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	B-Disease
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
molecular	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
colorectal	NN	O	B-Disease
tumor	NN	O	I-Disease
growth	NN	O	O
might	NN	O	O
be	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
driven	NN	O	O
by	NN	O	O
selection	NN	O	O
for	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
mutation	NN	O	O
cluster	NN	O	O
region	NN	O	O
.	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
interacts	NN	O	O
with	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
histone	NN	O	O
deacetylase	NN	O	O
complex	NN	O	O
.	NN	O	O

Germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
tumor	NN	O	O
-	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

BRCA1	NN	O	O
contains	NN	O	O
a	NN	O	O
carboxyl	NN	O	O
-	NN	O	O
terminal	NN	O	O
domain	NN	O	O
(	NN	O	O
BRCT	NN	O	O
)	NN	O	O
that	NN	O	O
is	NN	O	O
shared	NN	O	O
with	NN	O	O
several	NN	O	O
other	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
maintaining	NN	O	O
genome	NN	O	O
integrity	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
isolate	NN	O	O
proteins	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
BRCT	NN	O	O
domain	NN	O	O
.	NN	O	O

Purified	NN	O	O
BRCT	NN	O	O
polypeptide	NN	O	O
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
probe	NN	O	O
to	NN	O	O
screen	NN	O	O
a	NN	O	O
human	NN	O	O
placenta	NN	O	O
cDNA	NN	O	O
expression	NN	O	O
library	NN	O	O
by	NN	O	O
Far	NN	O	O
Western	NN	O	O
analysis	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
interacts	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
the	NN	O	O
Rb	NN	O	O
-	NN	O	O
binding	NN	O	O
proteins	NN	O	O
,	NN	O	O
RbAp46	NN	O	O
and	NN	O	O
RbAp48	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
with	NN	O	O
Rb	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
BRCT	NN	O	O
domain	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
histone	NN	O	O
deacetylases	NN	O	O
HDAC1	NN	O	O
and	NN	O	O
HDAC2	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
interacts	NN	O	O
with	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
histone	NN	O	O
deacetylase	NN	O	O
complex	NN	O	O
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
multiple	NN	O	O
processes	NN	O	O
such	NN	O	O
as	NN	O	O
transcription	NN	O	O
,	NN	O	O
DNA	NN	O	O
repair	NN	O	O
,	NN	O	O
and	NN	O	O
recombination	NN	O	O
.	NN	O	O
.	NN	O	O

Combined	NN	O	B-Disease
genetic	NN	O	I-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
C6	NN	O	I-Disease
and	NN	O	I-Disease
C7	NN	O	I-Disease
in	NN	O	O
man	NN	O	O
.	NN	O	O

By	NN	O	O
routine	NN	O	O
screening	NN	O	O
of	NN	O	O
sera	NN	O	O
,	NN	O	O
a	NN	O	O
subject	NN	O	O
was	NN	O	O
discovered	NN	O	O
who	NN	O	O
showed	NN	O	O
a	NN	O	O
sub	NN	O	B-Disease
-	NN	O	I-Disease
total	NN	O	I-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
C6	NN	O	I-Disease
and	NN	O	I-Disease
C7	NN	O	I-Disease
.	NN	O	O

No	NN	O	O
clinical	NN	O	O
disease	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
this	NN	O	O
deficiency	NN	O	O
which	NN	O	O
was	NN	O	O
transmitted	NN	O	O
through	NN	O	O
the	NN	O	O
subjects	NN	O	O
family	NN	O	O
as	NN	O	O
a	NN	O	O
single	NN	O	O
genetic	NN	O	O
characteristic	NN	O	O
,	NN	O	O
the	NN	O	O
C6	NN	O	B-Disease
deficiency	NN	O	I-Disease
being	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
silent	NN	O	O
allele	NN	O	O
at	NN	O	O
the	NN	O	O
structural	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
propositus	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
low	NN	O	O
quantities	NN	O	O
of	NN	O	O
an	NN	O	O
abnormal	NN	O	O
C6	NN	O	O
which	NN	O	O
was	NN	O	O
both	NN	O	O
antigenically	NN	O	O
deficient	NN	O	O
and	NN	O	O
smaller	NN	O	O
in	NN	O	O
size	NN	O	O
than	NN	O	O
normal	NN	O	O
C6	NN	O	O
(	NN	O	O
110	NN	O	O
,	NN	O	O
000	NN	O	O
daltons	NN	O	O
compared	NN	O	O
with	NN	O	O
140	NN	O	O
,	NN	O	O
000	NN	O	O
daltons	NN	O	O
)	NN	O	O
and	NN	O	O
small	NN	O	O
quantities	NN	O	O
of	NN	O	O
apparently	NN	O	O
normal	NN	O	O
C7	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
the	NN	O	O
most	NN	O	O
likely	NN	O	O
explanation	NN	O	O
for	NN	O	O
this	NN	O	O
defect	NN	O	O
is	NN	O	O
that	NN	O	O
the	NN	O	O
subject	NN	O	O
has	NN	O	O
a	NN	O	O
structural	NN	O	O
mutation	NN	O	O
in	NN	O	O
his	NN	O	O
C6	NN	O	O
gene	NN	O	O
which	NN	O	O
produces	NN	O	O
hyopsynthesis	NN	O	O
not	NN	O	O
only	NN	O	O
of	NN	O	O
C6	NN	O	O
but	NN	O	O
also	NN	O	O
of	NN	O	O
the	NN	O	O
closely	NN	O	O
linked	NN	O	O
gene	NN	O	O
for	NN	O	O
C7	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
C6	NN	O	O
and	NN	O	O
C7	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
single	NN	O	O
genetic	NN	O	O
unit	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
primary	NN	O	O
transcript	NN	O	O
copied	NN	O	O
from	NN	O	O
the	NN	O	O
genome	NN	O	O
includes	NN	O	O
information	NN	O	O
for	NN	O	O
both	NN	O	O
proteins	NN	O	O
.	NN	O	O
.	NN	O	O

Changes	NN	O	O
at	NN	O	O
P183	NN	O	O
of	NN	O	O
emerin	NN	O	O
weaken	NN	O	O
its	NN	O	O
protein	NN	O	O
-	NN	O	O
protein	NN	O	O
interactions	NN	O	O
resulting	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Emery	NN	O	B-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EDMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
early	NN	O	O
contractures	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
elbows	NN	O	I-Disease
,	NN	O	I-Disease
Achilles	NN	O	I-Disease
tendons	NN	O	I-Disease
and	NN	O	I-Disease
spine	NN	O	I-Disease
,	NN	O	O
slowly	NN	O	O
progressive	NN	O	O
muscle	NN	O	B-Disease
wasting	NN	O	I-Disease
and	NN	O	O
weakness	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
cardiomyopathy	NN	O	B-Disease
associated	NN	O	O
with	NN	O	O
cardiac	NN	O	B-Disease
conduction	NN	O	I-Disease
defects	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
emerin	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
Xq28	NN	O	O
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
34	NN	O	O
-	NN	O	O
kDa	NN	O	O
serine	NN	O	O
-	NN	O	O
rich	NN	O	O
protein	NN	O	O
,	NN	O	O
emerin	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
envelope	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
tissues	NN	O	O
,	NN	O	O
including	NN	O	O
skeletal	NN	O	O
and	NN	O	O
cardiac	NN	O	O
muscle	NN	O	O
.	NN	O	O

Mutations	NN	O	O
spanning	NN	O	O
the	NN	O	O
emerin	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
EDMD	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
present	NN	O	O
here	NN	O	O
the	NN	O	O
effect	NN	O	O
,	NN	O	O
on	NN	O	O
emerin	NN	O	O
protein	NN	O	O
expression	NN	O	O
,	NN	O	O
of	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
identified	NN	O	O
in	NN	O	O
unrelated	NN	O	O
EDMD	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

These	NN	O	O
alterations	NN	O	O
predict	NN	O	O
the	NN	O	O
replacement	NN	O	O
of	NN	O	O
a	NN	O	O
proline	NN	O	O
residue	NN	O	O
at	NN	O	O
position	NN	O	O
183	NN	O	O
with	NN	O	O
either	NN	O	O
a	NN	O	O
histidine	NN	O	O
or	NN	O	O
a	NN	O	O
threonine	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
analysis	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
mobility	NN	O	O
and	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
emerin	NN	O	O
are	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
emerin	NN	O	O
,	NN	O	O
but	NN	O	O
that	NN	O	O
they	NN	O	O
have	NN	O	O
weakened	NN	O	O
interactions	NN	O	O
with	NN	O	O
nuclear	NN	O	O
lamina	NN	O	O
components	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
with	NN	O	O
the	NN	O	O
usual	NN	O	O
EDMD	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
P183	NN	O	O
missense	NN	O	O
mutations	NN	O	O
have	NN	O	O
a	NN	O	O
later	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
of	NN	O	O
first	NN	O	O
symptoms	NN	O	O
,	NN	O	O
elbow	NN	O	B-Disease
contractures	NN	O	I-Disease
,	NN	O	O
ankle	NN	O	B-Disease
contractures	NN	O	I-Disease
,	NN	O	O
upper	NN	O	B-Disease
limb	NN	O	I-Disease
weakness	NN	O	I-Disease
and	NN	O	O
lower	NN	O	B-Disease
limb	NN	O	I-Disease
weakness	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
difference	NN	O	O
for	NN	O	O
the	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
of	NN	O	O
cardiac	NN	O	B-Disease
involvement	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
protein	NN	O	O
studies	NN	O	O
on	NN	O	O
patients	NN	O	O
with	NN	O	O
missense	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
EDMD	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
proline	NN	O	O
183	NN	O	O
for	NN	O	O
the	NN	O	O
proper	NN	O	O
structure	NN	O	O
/	NN	O	O
function	NN	O	O
of	NN	O	O
emerin	NN	O	O
.	NN	O	O
.	NN	O	O

Microdeletions	NN	O	O
at	NN	O	O
chromosome	NN	O	O
bands	NN	O	O
1q32	NN	O	O
-	NN	O	O
q41	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
Van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
VWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
disorder	NN	O	I-Disease
comprising	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
and	NN	O	O
/	NN	O	O
or	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
and	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
reported	NN	O	O
previously	NN	O	O
a	NN	O	O
family	NN	O	O
whose	NN	O	O
underlying	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
500	NN	O	O
-	NN	O	O
800	NN	O	O
kb	NN	O	O
deletion	NN	O	O
localized	NN	O	O
to	NN	O	O
chromosome	NN	O	O
bands	NN	O	O
1q32	NN	O	O
-	NN	O	O
q41	NN	O	O
[	NN	O	O
Sander	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1994	NN	O	O
Hum	NN	O	O
Mol	NN	O	O
Genet	NN	O	O
3	NN	O	O
576	NN	O	O
-	NN	O	O
578	NN	O	O
]	NN	O	O
.	NN	O	O

Along	NN	O	O
with	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
/	NN	O	I-Disease
palate	NN	O	I-Disease
and	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
,	NN	O	O
affected	NN	O	O
relatives	NN	O	O
exhibit	NN	O	O
developmental	NN	O	B-Disease
delays	NN	O	I-Disease
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
nearby	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
disrupted	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
localize	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
we	NN	O	O
searched	NN	O	O
for	NN	O	O
other	NN	O	O
deletions	NN	O	O
that	NN	O	O
cause	NN	O	O
VWS	NN	O	B-Disease
.	NN	O	O

An	NN	O	O
allele	NN	O	O
loss	NN	O	O
assay	NN	O	O
was	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
novel	NN	O	O
highly	NN	O	O
polymorphic	NN	O	O
marker	NN	O	O
,	NN	O	O
D1S3753	NN	O	O
.	NN	O	O

From	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
37	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
an	NN	O	O
allele	NN	O	O
loss	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
deletion	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
the	NN	O	O
phenotype	NN	O	O
in	NN	O	O
three	NN	O	O
generations	NN	O	O
of	NN	O	O
affected	NN	O	O
individuals	NN	O	O
was	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
cardinal	NN	O	O
signs	NN	O	O
of	NN	O	O
VWS	NN	O	B-Disease
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
new	NN	O	O
deletion	NN	O	O
showed	NN	O	O
that	NN	O	O
it	NN	O	O
extended	NN	O	O
0	NN	O	O
.	NN	O	O

2	NN	O	O
-	NN	O	O
1	NN	O	O
Mb	NN	O	O
beyond	NN	O	O
the	NN	O	O
proximal	NN	O	O
breakpoint	NN	O	O
for	NN	O	O
the	NN	O	O
deletion	NN	O	O
described	NN	O	O
previously	NN	O	O
.	NN	O	O

No	NN	O	O
deletions	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
seven	NN	O	O
cases	NN	O	O
of	NN	O	O
popliteal	NN	O	B-Disease
pterygia	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
76	NN	O	O
cases	NN	O	O
of	NN	O	O
mixed	NN	O	O
syndromic	NN	O	B-Disease
forms	NN	O	I-Disease
of	NN	O	I-Disease
cleft	NN	O	I-Disease
lip	NN	O	I-Disease
and	NN	O	I-Disease
palate	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
178	NN	O	O
cases	NN	O	O
of	NN	O	O
nonsyndromic	NN	O	B-Disease
cleft	NN	O	I-Disease
lip	NN	O	I-Disease
and	NN	O	I-Disease
palate	NN	O	I-Disease
.	NN	O	O

Splicing	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
gene	NN	O	O
,	NN	O	O
ATM	NN	O	O
:	NN	O	O
underlying	NN	O	O
mutations	NN	O	O
and	NN	O	O
consequences	NN	O	O
.	NN	O	O

Mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
defective	NN	O	O
splicing	NN	O	O
constitute	NN	O	O
a	NN	O	O
significant	NN	O	O
proportion	NN	O	O
(	NN	O	O
30	NN	O	O
/	NN	O	O
62	NN	O	O
[	NN	O	O
48	NN	O	O
%	NN	O	O
]	NN	O	O
)	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
series	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
AT	NN	O	B-Disease
)	NN	O	O
that	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
the	NN	O	O
protein	NN	O	O
-	NN	O	O
truncation	NN	O	O
assay	NN	O	O
followed	NN	O	O
by	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
.	NN	O	O

Fewer	NN	O	O
than	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
splicing	NN	O	O
mutations	NN	O	O
involved	NN	O	O
the	NN	O	O
canonical	NN	O	O
AG	NN	O	O
splice	NN	O	O
-	NN	O	O
acceptor	NN	O	O
site	NN	O	O
or	NN	O	O
GT	NN	O	O
splice	NN	O	O
-	NN	O	O
donor	NN	O	O
site	NN	O	O
.	NN	O	O

A	NN	O	O
higher	NN	O	O
percentage	NN	O	O
of	NN	O	O
mutations	NN	O	O
occurred	NN	O	O
at	NN	O	O
less	NN	O	O
stringently	NN	O	O
conserved	NN	O	O
sites	NN	O	O
,	NN	O	O
including	NN	O	O
silent	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
last	NN	O	O
nucleotide	NN	O	O
of	NN	O	O
exons	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
nucleotides	NN	O	O
other	NN	O	O
than	NN	O	O
the	NN	O	O
conserved	NN	O	O
AG	NN	O	O
and	NN	O	O
GT	NN	O	O
in	NN	O	O
the	NN	O	O
consensus	NN	O	O
splice	NN	O	O
sites	NN	O	O
,	NN	O	O
and	NN	O	O
creation	NN	O	O
of	NN	O	O
splice	NN	O	O
-	NN	O	O
acceptor	NN	O	O
or	NN	O	O
splice	NN	O	O
-	NN	O	O
donor	NN	O	O
sites	NN	O	O
in	NN	O	O
either	NN	O	O
introns	NN	O	O
or	NN	O	O
exons	NN	O	O
.	NN	O	O

These	NN	O	O
splicing	NN	O	O
mutations	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
consequences	NN	O	O
,	NN	O	O
including	NN	O	O
exon	NN	O	O
skipping	NN	O	O
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
degree	NN	O	O
,	NN	O	O
intron	NN	O	O
retention	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
sites	NN	O	O
,	NN	O	O
or	NN	O	O
creation	NN	O	O
of	NN	O	O
new	NN	O	O
splice	NN	O	O
sites	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
5	NN	O	O
of	NN	O	O
12	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
and	NN	O	O
1	NN	O	O
missense	NN	O	O
mutation	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
cDNA	NN	O	O
of	NN	O	O
the	NN	O	O
exons	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
mutations	NN	O	O
occurred	NN	O	O
.	NN	O	O

No	NN	O	O
ATM	NN	O	O
protein	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
western	NN	O	O
blotting	NN	O	O
in	NN	O	O
any	NN	O	O
AT	NN	O	B-Disease
cell	NN	O	O
line	NN	O	O
in	NN	O	O
which	NN	O	O
splicing	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Several	NN	O	O
cases	NN	O	O
of	NN	O	O
exon	NN	O	O
skipping	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	O
controls	NN	O	O
and	NN	O	O
patients	NN	O	O
for	NN	O	O
whom	NN	O	O
no	NN	O	O
underlying	NN	O	O
defect	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
in	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
,	NN	O	O
suggesting	NN	O	O
caution	NN	O	O
in	NN	O	O
the	NN	O	O
interpretation	NN	O	O
of	NN	O	O
exon	NN	O	O
deletions	NN	O	O
observed	NN	O	O
in	NN	O	O
ATM	NN	O	O
cDNA	NN	O	O
when	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
accompanying	NN	O	O
identification	NN	O	O
of	NN	O	O
genomic	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Alpha	NN	O	O
-	NN	O	O
cardiac	NN	O	O
actin	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
disease	NN	O	O
gene	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
hypertrophic	NN	O	I-Disease
cardiomyopathy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
identified	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
cardiac	NN	O	O
actin	NN	O	O
gene	NN	O	O
(	NN	O	O
ACTC	NN	O	O
)	NN	O	O
as	NN	O	O
a	NN	O	O
novel	NN	O	O
disease	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
pedigree	NN	O	O
suffering	NN	O	O
from	NN	O	O
familial	NN	O	B-Disease
hypertrophic	NN	O	I-Disease
cardiomyopathy	NN	O	I-Disease
(	NN	O	O
FHC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analyses	NN	O	O
excluded	NN	O	O
all	NN	O	O
the	NN	O	O
previously	NN	O	O
reported	NN	O	O
FHC	NN	O	B-Disease
loci	NN	O	O
as	NN	O	O
possible	NN	O	O
disease	NN	O	O
loci	NN	O	O
in	NN	O	O
the	NN	O	O
family	NN	O	O
studied	NN	O	O
,	NN	O	O
with	NN	O	O
lod	NN	O	O
scores	NN	O	O
varying	NN	O	O
between	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O

5	NN	O	O
and	NN	O	O
-	NN	O	O
6	NN	O	O
.	NN	O	O

0	NN	O	O
0	NN	O	O
.	NN	O	O

Further	NN	O	O
linkage	NN	O	O
analyses	NN	O	O
of	NN	O	O
plausible	NN	O	O
candidate	NN	O	O
genes	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
adult	NN	O	O
human	NN	O	O
heart	NN	O	O
identified	NN	O	O
ACTC	NN	O	O
as	NN	O	O
the	NN	O	O
most	NN	O	O
likely	NN	O	O
disease	NN	O	O
gene	NN	O	O
,	NN	O	O
showing	NN	O	O
a	NN	O	O
maximal	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
3	NN	O	O
.	NN	O	O

6	NN	O	O
6	NN	O	O
.	NN	O	O

Mutation	NN	O	O
analysis	NN	O	O
of	NN	O	O
ACTC	NN	O	O
revealed	NN	O	O
an	NN	O	O
Ala295Ser	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
close	NN	O	O
to	NN	O	O
2	NN	O	O
missense	NN	O	O
mutations	NN	O	O
recently	NN	O	O
described	NN	O	O
to	NN	O	O
cause	NN	O	O
the	NN	O	O
inherited	NN	O	O
form	NN	O	O
of	NN	O	O
idiopathic	NN	O	B-Disease
dilated	NN	O	I-Disease
cardiomyopathy	NN	O	I-Disease
(	NN	O	O
IDC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Overgrowth	NN	O	B-Disease
of	NN	O	I-Disease
oral	NN	O	I-Disease
mucosa	NN	O	I-Disease
and	NN	O	I-Disease
facial	NN	O	I-Disease
skin	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
novel	NN	O	O
feature	NN	O	O
of	NN	O	O
aspartylglucosaminuria	NN	O	B-Disease
.	NN	O	O

Aspartylglucosaminuria	NN	O	B-Disease
(	NN	O	O
AGU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
lysosomal	NN	O	B-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
aspartylglucosaminidase	NN	O	I-Disease
(	NN	O	O
AGA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
main	NN	O	O
symptom	NN	O	O
is	NN	O	O
progressive	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
spectrum	NN	O	O
of	NN	O	O
different	NN	O	O
mutations	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
this	NN	O	O
disease	NN	O	O
,	NN	O	O
one	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
Cys163Ser	NN	O	O
)	NN	O	O
being	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
Finnish	NN	O	O
cases	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
examine	NN	O	O
66	NN	O	O
Finnish	NN	O	O
AGU	NN	O	B-Disease
patients	NN	O	O
for	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
oral	NN	O	O
mucosa	NN	O	O
and	NN	O	O
44	NN	O	O
of	NN	O	O
these	NN	O	O
for	NN	O	O
changes	NN	O	O
in	NN	O	O
facial	NN	O	O
skin	NN	O	O
.	NN	O	O

Biopsy	NN	O	O
specimens	NN	O	O
of	NN	O	O
16	NN	O	O
oral	NN	O	O
lesions	NN	O	O
,	NN	O	O
12	NN	O	O
of	NN	O	O
them	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
teeth	NN	O	O
,	NN	O	O
plus	NN	O	O
two	NN	O	O
facial	NN	O	B-Disease
lesions	NN	O	I-Disease
were	NN	O	O
studied	NN	O	O
histologically	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
staining	NN	O	O
for	NN	O	O
AGA	NN	O	O
was	NN	O	O
performed	NN	O	O
on	NN	O	O
15	NN	O	O
oral	NN	O	O
specimens	NN	O	O
.	NN	O	O

Skin	NN	O	O
was	NN	O	O
seborrhoeic	NN	O	O
in	NN	O	O
adolescent	NN	O	O
and	NN	O	O
adult	NN	O	O
patients	NN	O	O
,	NN	O	O
with	NN	O	O
erythema	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
facial	NN	O	I-Disease
skin	NN	O	I-Disease
already	NN	O	O
common	NN	O	O
in	NN	O	O
childhood	NN	O	O
.	NN	O	O

Of	NN	O	O
44	NN	O	O
patients	NN	O	O
,	NN	O	O
nine	NN	O	O
(	NN	O	O
20	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
facial	NN	O	B-Disease
angiofibromas	NN	O	I-Disease
,	NN	O	O
tumours	NN	O	B-Disease
primarily	NN	O	O
occurring	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
tuberous	NN	O	B-Disease
sclerosis	NN	O	I-Disease
.	NN	O	O

Oedemic	NN	O	B-Disease
buccal	NN	O	I-Disease
mucosa	NN	O	I-Disease
(	NN	O	O
leucoedema	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
gingival	NN	O	B-Disease
overgrowths	NN	O	I-Disease
were	NN	O	O
more	NN	O	O
frequent	NN	O	O
in	NN	O	O
AGU	NN	O	B-Disease
patients	NN	O	O
than	NN	O	O
in	NN	O	O
controls	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
.	NN	O	O

Of	NN	O	O
16	NN	O	O
oral	NN	O	B-Disease
mucosal	NN	O	I-Disease
lesions	NN	O	I-Disease
studied	NN	O	O
histologically	NN	O	O
,	NN	O	O
15	NN	O	O
represented	NN	O	O
fibroepithelial	NN	O	B-Disease
or	NN	O	I-Disease
epithelial	NN	O	I-Disease
hyperplasias	NN	O	I-Disease
and	NN	O	O
were	NN	O	O
reactive	NN	O	O
in	NN	O	O
nature	NN	O	O
.	NN	O	O

Cytoplasmic	NN	O	O
vacuolisation	NN	O	O
was	NN	O	O
evident	NN	O	O
in	NN	O	O
four	NN	O	O
.	NN	O	O

Immunohistochemically	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
AGA	NN	O	O
in	NN	O	O
AGU	NN	O	B-Disease
patients	NN	O	O
mucosal	NN	O	B-Disease
lesions	NN	O	I-Disease
did	NN	O	O
not	NN	O	O
differ	NN	O	O
from	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
corresponding	NN	O	O
lesions	NN	O	O
of	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
mucosal	NN	O	B-Disease
overgrowth	NN	O	I-Disease
in	NN	O	O
AGU	NN	O	B-Disease
patients	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
directly	NN	O	O
associated	NN	O	O
with	NN	O	O
lysosomal	NN	O	O
storage	NN	O	O
or	NN	O	O
with	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
AGA	NN	O	O
expression	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
germline	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
seven	NN	O	O
novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
OCRL1	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
oculocerebrorenal	NN	O	B-Disease
syndrome	NN	O	I-Disease
of	NN	O	I-Disease
Lowe	NN	O	I-Disease
(	NN	O	O
OCRL	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
major	NN	O	O
abnormalities	NN	O	B-Disease
of	NN	O	I-Disease
eyes	NN	O	I-Disease
,	NN	O	I-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
,	NN	O	I-Disease
and	NN	O	I-Disease
kidneys	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
OCRL1	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

OCRL1	NN	O	O
encodes	NN	O	O
a	NN	O	O
phosphatidylinositol	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
biphosphate	NN	O	O
(	NN	O	O
PtdIns	NN	O	O
[	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
]	NN	O	O
P2	NN	O	O
)	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
OCRL1	NN	O	O
gene	NN	O	O
in	NN	O	O
eight	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
with	NN	O	O
OCRL	NN	O	B-Disease
and	NN	O	O
have	NN	O	O
found	NN	O	O
seven	NN	O	O
new	NN	O	O
mutations	NN	O	O
and	NN	O	O
one	NN	O	O
recurrent	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
.	NN	O	O

Among	NN	O	O
the	NN	O	O
new	NN	O	O
mutations	NN	O	O
,	NN	O	O
two	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
(	NN	O	O
R317X	NN	O	O
and	NN	O	O
E558X	NN	O	O
)	NN	O	O
and	NN	O	O
three	NN	O	O
other	NN	O	O
frameshift	NN	O	O
mutations	NN	O	O
caused	NN	O	O
premature	NN	O	O
termination	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

A	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
R483G	NN	O	O
,	NN	O	O
was	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
highly	NN	O	O
conserved	NN	O	O
PtdIns	NN	O	O
(	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
)	NN	O	O
P2	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
domain	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
one	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
,	NN	O	O
2799delC	NN	O	O
,	NN	O	O
modifies	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
part	NN	O	O
of	NN	O	O
OCRL1	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
extension	NN	O	O
of	NN	O	O
six	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
missense	NN	O	O
mutations	NN	O	O
are	NN	O	O
located	NN	O	O
in	NN	O	O
exon	NN	O	O
15	NN	O	O
,	NN	O	O
and	NN	O	O
52	NN	O	O
%	NN	O	O
of	NN	O	O
all	NN	O	O
mutations	NN	O	O
cluster	NN	O	O
in	NN	O	O
exons	NN	O	O
11	NN	O	O
-	NN	O	O
15	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
identified	NN	O	O
two	NN	O	O
new	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
for	NN	O	O
the	NN	O	O
OCRL1	NN	O	O
locus	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
detected	NN	O	O
a	NN	O	O
germline	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
has	NN	O	O
direct	NN	O	O
implications	NN	O	O
for	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
of	NN	O	O
Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
families	NN	O	O
.	NN	O	O
.	NN	O	O

MEFV	NN	O	O
-	NN	O	O
Gene	NN	O	O
analysis	NN	O	O
in	NN	O	O
armenian	NN	O	O
patients	NN	O	O
with	NN	O	O
Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
:	NN	O	O
diagnostic	NN	O	O
value	NN	O	O
and	NN	O	O
unfavorable	NN	O	O
renal	NN	O	O
prognosis	NN	O	O
of	NN	O	O
the	NN	O	O
M694V	NN	O	O
homozygous	NN	O	O
genotype	NN	O	O
-	NN	O	O
genetic	NN	O	O
and	NN	O	O
therapeutic	NN	O	O
implications	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
recessively	NN	O	B-Disease
inherited	NN	O	I-Disease
disorder	NN	O	I-Disease
that	NN	O	O
is	NN	O	O
common	NN	O	O
in	NN	O	O
patients	NN	O	O
of	NN	O	O
Armenian	NN	O	O
ancestry	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
,	NN	O	O
its	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
made	NN	O	O
only	NN	O	O
retrospectively	NN	O	O
,	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
exclusion	NN	O	O
,	NN	O	O
based	NN	O	O
entirely	NN	O	O
on	NN	O	O
nonspecific	NN	O	O
clinical	NN	O	O
signs	NN	O	O
that	NN	O	O
result	NN	O	O
from	NN	O	O
serosal	NN	O	B-Disease
inflammation	NN	O	I-Disease
and	NN	O	O
that	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
unnecessary	NN	O	O
surgery	NN	O	O
.	NN	O	O

Renal	NN	O	B-Disease
amyloidosis	NN	O	I-Disease
,	NN	O	O
prevented	NN	O	O
by	NN	O	O
colchicine	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
severe	NN	O	O
complication	NN	O	O
of	NN	O	O
FMF	NN	O	B-Disease
,	NN	O	O
a	NN	O	O
disorder	NN	O	O
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
MEFV	NN	O	O
gene	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
diagnostic	NN	O	O
and	NN	O	O
prognostic	NN	O	O
value	NN	O	O
of	NN	O	O
MEFV	NN	O	O
-	NN	O	O
gene	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
90	NN	O	O
Armenian	NN	O	O
FMF	NN	O	B-Disease
patients	NN	O	O
from	NN	O	O
77	NN	O	O
unrelated	NN	O	O
families	NN	O	O
that	NN	O	O
were	NN	O	O
not	NN	O	O
selected	NN	O	O
through	NN	O	O
genetic	NN	O	O
-	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
.	NN	O	O

Eight	NN	O	O
mutations	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
(	NN	O	O
R408Q	NN	O	O
)	NN	O	O
is	NN	O	O
new	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
93	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
163	NN	O	O
independent	NN	O	O
FMF	NN	O	B-Disease
alleles	NN	O	O
,	NN	O	O
with	NN	O	O
both	NN	O	O
FMF	NN	O	B-Disease
alleles	NN	O	O
identified	NN	O	O
in	NN	O	O
89	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
several	NN	O	O
instances	NN	O	O
,	NN	O	O
family	NN	O	O
studies	NN	O	O
provided	NN	O	O
molecular	NN	O	O
evidence	NN	O	O
for	NN	O	O
pseudodominant	NN	O	O
transmission	NN	O	O
and	NN	O	O
incomplete	NN	O	O
penetrance	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
M694V	NN	O	O
homozygous	NN	O	O
genotype	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
higher	NN	O	O
prevalence	NN	O	O
of	NN	O	O
renal	NN	O	B-Disease
amyloidosis	NN	O	I-Disease
and	NN	O	O
arthritis	NN	O	B-Disease
,	NN	O	O
compared	NN	O	O
with	NN	O	O
other	NN	O	O
genotypes	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
0002	NN	O	O
and	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
006	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
demonstration	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
diagnostic	NN	O	O
and	NN	O	O
prognostic	NN	O	O
value	NN	O	O
of	NN	O	O
MEFV	NN	O	O
analysis	NN	O	O
and	NN	O	O
particular	NN	O	O
modes	NN	O	O
of	NN	O	O
inheritance	NN	O	O
should	NN	O	O
lead	NN	O	O
to	NN	O	O
new	NN	O	O
ways	NN	O	O
for	NN	O	O
management	NN	O	O
of	NN	O	O
FMF	NN	O	O
-	NN	O	O
including	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
and	NN	O	O
therapeutic	NN	O	O
decisions	NN	O	O
in	NN	O	O
affected	NN	O	O
families	NN	O	O
.	NN	O	O

Noninvasive	NN	O	O
test	NN	O	O
for	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
using	NN	O	O
hair	NN	O	O
root	NN	O	O
analysis	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
repeat	NN	O	O
-	NN	O	O
amplification	NN	O	O
mechanism	NN	O	O
causing	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
led	NN	O	O
to	NN	O	O
development	NN	O	O
of	NN	O	O
reliable	NN	O	O
DNA	NN	O	O
-	NN	O	O
based	NN	O	O
diagnostic	NN	O	O
methods	NN	O	O
,	NN	O	O
including	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
and	NN	O	O
PCR	NN	O	O
.	NN	O	O

Both	NN	O	O
methods	NN	O	O
are	NN	O	O
performed	NN	O	O
on	NN	O	O
DNA	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
cells	NN	O	O
and	NN	O	O
measure	NN	O	O
the	NN	O	O
repeat	NN	O	O
size	NN	O	O
in	NN	O	O
FMR1	NN	O	O
.	NN	O	O

Using	NN	O	O
an	NN	O	O
immunocytochemical	NN	O	O
technique	NN	O	O
on	NN	O	O
blood	NN	O	O
smears	NN	O	O
,	NN	O	O
we	NN	O	O
recently	NN	O	O
developed	NN	O	O
a	NN	O	O
novel	NN	O	O
test	NN	O	O
for	NN	O	O
identification	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
method	NN	O	O
,	NN	O	O
also	NN	O	O
called	NN	O	O
"	NN	O	O
antibody	NN	O	O
test	NN	O	O
,	NN	O	O
"	NN	O	O
uses	NN	O	O
monoclonal	NN	O	O
antibodies	NN	O	O
against	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
product	NN	O	O
(	NN	O	O
FMRP	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
based	NN	O	O
on	NN	O	O
absence	NN	O	O
of	NN	O	O
FMRP	NN	O	O
in	NN	O	O
patients	NN	O	O
cells	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
a	NN	O	O
new	NN	O	O
diagnostic	NN	O	O
test	NN	O	O
to	NN	O	O
identify	NN	O	O
male	NN	O	O
patients	NN	O	O
with	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
lack	NN	O	O
of	NN	O	O
FMRP	NN	O	O
in	NN	O	O
their	NN	O	O
hair	NN	O	O
roots	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
FMRP	NN	O	O
in	NN	O	O
hair	NN	O	O
roots	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
FMRP	NN	O	O
-	NN	O	O
specific	NN	O	O
antibody	NN	O	O
test	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
FMRP	NN	O	O
-	NN	O	O
expressing	NN	O	O
hair	NN	O	O
roots	NN	O	O
in	NN	O	O
controls	NN	O	O
and	NN	O	O
in	NN	O	O
male	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
patients	NN	O	O
was	NN	O	O
determined	NN	O	O
.	NN	O	O

Control	NN	O	O
individuals	NN	O	O
showed	NN	O	O
clear	NN	O	O
expression	NN	O	O
of	NN	O	O
FMRP	NN	O	O
in	NN	O	O
nearly	NN	O	O
every	NN	O	O
hair	NN	O	O
root	NN	O	O
,	NN	O	O
whereas	NN	O	O
male	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
patients	NN	O	O
lacked	NN	O	O
expression	NN	O	O
of	NN	O	O
FMRP	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
their	NN	O	O
hair	NN	O	O
roots	NN	O	O
.	NN	O	O

Mentally	NN	O	B-Disease
retarded	NN	O	I-Disease
female	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
full	NN	O	O
mutation	NN	O	O
showed	NN	O	O
FMRP	NN	O	O
expression	NN	O	O
in	NN	O	O
only	NN	O	O
some	NN	O	O
of	NN	O	O
their	NN	O	O
hair	NN	O	O
roots	NN	O	O
(	NN	O	O
<	NN	O	O
55	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
overlap	NN	O	O
with	NN	O	O
normal	NN	O	O
female	NN	O	O
controls	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
advantages	NN	O	O
of	NN	O	O
this	NN	O	O
test	NN	O	O
are	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
plucking	NN	O	O
of	NN	O	O
hair	NN	O	O
follicles	NN	O	O
does	NN	O	O
no	NN	O	O
appreciable	NN	O	O
harm	NN	O	O
to	NN	O	O
the	NN	O	O
mentally	NN	O	B-Disease
retarded	NN	O	I-Disease
patient	NN	O	O
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
hairs	NN	O	O
can	NN	O	O
be	NN	O	O
sent	NN	O	O
in	NN	O	O
a	NN	O	O
simple	NN	O	O
envelope	NN	O	O
to	NN	O	O
a	NN	O	O
diagnostic	NN	O	O
center	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
test	NN	O	O
is	NN	O	O
available	NN	O	O
within	NN	O	O
5	NN	O	O
h	NN	O	O
of	NN	O	O
plucking	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
test	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
identify	NN	O	O
two	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
patients	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
the	NN	O	O
full	NN	O	O
mutation	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
isolated	NN	O	O
from	NN	O	O
blood	NN	O	O
cells	NN	O	O
.	NN	O	O
.	NN	O	O

In	NN	O	O
Swedish	NN	O	O
families	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
HPC1	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
families	NN	O	O
with	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
clusters	NN	O	O
in	NN	O	O
some	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
estimated	NN	O	O
5	NN	O	O
%	NN	O	O
-	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
all	NN	O	O
cases	NN	O	O
are	NN	O	O
estimated	NN	O	O
to	NN	O	O
result	NN	O	O
from	NN	O	O
inheritance	NN	O	O
of	NN	O	O
prostate	NN	O	O
cancer	NN	O	O
-	NN	O	O
susceptibility	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
region	NN	O	O
(	NN	O	O
HPC1	NN	O	O
)	NN	O	O
in	NN	O	O
91	NN	O	O
North	NN	O	O
American	NN	O	O
and	NN	O	O
Swedish	NN	O	O
families	NN	O	O
each	NN	O	O
with	NN	O	O
multiple	NN	O	O
cases	NN	O	O
of	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
(	NN	O	O
Smith	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
1996	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
we	NN	O	O
analyze	NN	O	O
40	NN	O	O
(	NN	O	O
12	NN	O	O
original	NN	O	O
and	NN	O	O
28	NN	O	O
newly	NN	O	O
identified	NN	O	O
)	NN	O	O
Swedish	NN	O	O
families	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
(	NN	O	O
HPC	NN	O	B-Disease
)	NN	O	O
that	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
40	NN	O	O
markers	NN	O	O
spanning	NN	O	O
a	NN	O	O
25	NN	O	O
-	NN	O	O
cM	NN	O	O
interval	NN	O	O
within	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
,	NN	O	O
have	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
complete	NN	O	O
set	NN	O	O
of	NN	O	O
families	NN	O	O
,	NN	O	O
a	NN	O	O
maximum	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

10	NN	O	O
was	NN	O	O
observed	NN	O	O
at	NN	O	O
D1S413	NN	O	O
(	NN	O	O
at	NN	O	O
a	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
[	NN	O	O
theta	NN	O	O
]	NN	O	O
of	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
maximum	NN	O	O
NPL	NN	O	O
(	NN	O	O
nonparametric	NN	O	O
linkage	NN	O	O
)	NN	O	O
Z	NN	O	O
score	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

64	NN	O	O
at	NN	O	O
D1S202	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
05	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
originated	NN	O	O
almost	NN	O	O
exclusively	NN	O	O
from	NN	O	O
the	NN	O	O
subset	NN	O	O
of	NN	O	O
12	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
(	NN	O	O
age	NN	O	O
<	NN	O	O
65	NN	O	O
years	NN	O	O
)	NN	O	O
families	NN	O	O
,	NN	O	O
which	NN	O	O
yielded	NN	O	O
a	NN	O	O
maximum	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

38	NN	O	O
at	NN	O	O
D1S413	NN	O	O
(	NN	O	O
straight	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
)	NN	O	O
and	NN	O	O
an	NN	O	O
NPL	NN	O	O
Z	NN	O	O
score	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

95	NN	O	O
at	NN	O	O
D1S422	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
.	NN	O	O

Estimates	NN	O	O
from	NN	O	O
heterogeneity	NN	O	O
tests	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
within	NN	O	O
Sweden	NN	O	O
,	NN	O	O
as	NN	O	O
many	NN	O	O
as	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
families	NN	O	O
had	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
HPC1	NN	O	O
region	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
HPC1	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
particularly	NN	O	O
those	NN	O	O
with	NN	O	O
an	NN	O	O
early	NN	O	O
age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
feline	NN	O	O
beta	NN	O	B-Disease
-	NN	O	I-Disease
glucuronidase	NN	O	I-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
of	NN	O	O
mucopolysaccharidosis	NN	O	B-Disease
VII	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
family	NN	O	O
of	NN	O	O
domestic	NN	O	O
cats	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
exhibited	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
abnormalities	NN	O	O
consistent	NN	O	O
with	NN	O	O
mucopolysaccharidosis	NN	O	B-Disease
VII	NN	O	I-Disease
,	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
lysosomal	NN	O	I-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
beta	NN	O	B-Disease
-	NN	O	I-Disease
glucuronidase	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

beta	NN	O	O
-	NN	O	O
Glucuronidase	NN	O	O
activity	NN	O	O
was	NN	O	O
undetectable	NN	O	O
in	NN	O	O
affected	NN	O	O
cat	NN	O	O
fibroblasts	NN	O	O
and	NN	O	O
restored	NN	O	O
by	NN	O	O
retroviral	NN	O	O
gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
rat	NN	O	O
beta	NN	O	O
-	NN	O	O
glucuronidase	NN	O	O
cDNA	NN	O	O
.	NN	O	O

beta	NN	O	O
-	NN	O	O
Glucuronidase	NN	O	O
mRNA	NN	O	O
was	NN	O	O
normal	NN	O	O
in	NN	O	O
affected	NN	O	O
cat	NN	O	O
testis	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

Normal	NN	O	O
feline	NN	O	O
beta	NN	O	O
-	NN	O	O
glucuronidase	NN	O	O
cDNA	NN	O	O
was	NN	O	O
cloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
,	NN	O	O
and	NN	O	O
amplified	NN	O	O
from	NN	O	O
affected	NN	O	O
cat	NN	O	O
fibroblasts	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
coupled	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
cat	NN	O	O
cDNA	NN	O	O
that	NN	O	O
predicted	NN	O	O
an	NN	O	O
E351K	NN	O	O
substitution	NN	O	O
,	NN	O	O
destroyed	NN	O	O
a	NN	O	O
BssSI	NN	O	O
site	NN	O	O
,	NN	O	O
and	NN	O	O
eliminated	NN	O	O
GUSB	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
in	NN	O	O
expression	NN	O	O
studies	NN	O	O
.	NN	O	O

Multiple	NN	O	O
species	NN	O	O
comparison	NN	O	O
and	NN	O	O
the	NN	O	O
crystal	NN	O	O
structure	NN	O	O
of	NN	O	O
human	NN	O	O
beta	NN	O	O
-	NN	O	O
glucuronidase	NN	O	O
indicated	NN	O	O
that	NN	O	O
E351	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
residue	NN	O	O
most	NN	O	O
likely	NN	O	O
essential	NN	O	O
in	NN	O	O
maintenance	NN	O	O
of	NN	O	O
the	NN	O	O
enzymes	NN	O	O
conformation	NN	O	O
.	NN	O	O

BssSI	NN	O	O
digestion	NN	O	O
of	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
products	NN	O	O
amplified	NN	O	O
from	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
indicated	NN	O	O
that	NN	O	O
affected	NN	O	O
cats	NN	O	O
were	NN	O	O
homozygous	NN	O	O
and	NN	O	O
cats	NN	O	O
with	NN	O	O
half	NN	O	O
-	NN	O	O
normal	NN	O	O
beta	NN	O	O
-	NN	O	O
glucuronidase	NN	O	O
activity	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
missense	NN	O	O
mutation	NN	O	O
.	NN	O	O

Carriers	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
manner	NN	O	O
produced	NN	O	O
affected	NN	O	O
kittens	NN	O	O
in	NN	O	O
prospective	NN	O	O
breedings	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
feline	NN	O	O
MPS	NN	O	B-Disease
VII	NN	O	I-Disease
breeding	NN	O	O
colony	NN	O	O
has	NN	O	O
been	NN	O	O
established	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
common	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
rearrangement	NN	O	B-Disease
disorders	NN	O	I-Disease
on	NN	O	O
chromosome	NN	O	O
22q11	NN	O	O
.	NN	O	O

The	NN	O	O
chromosome	NN	O	O
22q11	NN	O	O
region	NN	O	O
is	NN	O	O
susceptible	NN	O	O
to	NN	O	O
rearrangements	NN	O	O
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
anomaly	NN	O	I-Disease
disorders	NN	O	I-Disease
and	NN	O	O
malignant	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

Three	NN	O	O
congenital	NN	O	B-Disease
anomaly	NN	O	I-Disease
disorders	NN	O	I-Disease
,	NN	O	O
cat	NN	O	B-Disease
-	NN	O	I-Disease
eye	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
der	NN	O	B-Disease
(	NN	O	I-Disease
)	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
velo	NN	O	B-Disease
-	NN	O	I-Disease
cardio	NN	O	I-Disease
-	NN	O	I-Disease
facial	NN	O	I-Disease
syndrome	NN	O	I-Disease
/	NN	O	O
DiGeorge	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
tetrasomy	NN	O	O
,	NN	O	O
trisomy	NN	O	O
or	NN	O	O
monosomy	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
for	NN	O	O
part	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22q11	NN	O	O
.	NN	O	O

VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
syndrome	NN	O	O
associated	NN	O	O
with	NN	O	O
22q11	NN	O	O
rearrangements	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
there	NN	O	O
are	NN	O	O
particular	NN	O	O
regions	NN	O	O
on	NN	O	O
22q11	NN	O	O
that	NN	O	O
are	NN	O	O
prone	NN	O	O
to	NN	O	O
rearrangements	NN	O	O
,	NN	O	O
the	NN	O	O
deletion	NN	O	O
end	NN	O	O
-	NN	O	O
points	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
patients	NN	O	O
were	NN	O	O
defined	NN	O	O
by	NN	O	O
haplotype	NN	O	O
analysis	NN	O	O
.	NN	O	O

Most	NN	O	O
VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
patients	NN	O	O
have	NN	O	O
a	NN	O	O
similar	NN	O	O
3	NN	O	O
Mb	NN	O	O
deletion	NN	O	O
,	NN	O	O
some	NN	O	O
have	NN	O	O
a	NN	O	O
nested	NN	O	O
distal	NN	O	O
deletion	NN	O	O
breakpoint	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
1	NN	O	O
.	NN	O	O

5	NN	O	O
Mb	NN	O	O
deletion	NN	O	O
and	NN	O	O
a	NN	O	O
few	NN	O	O
rare	NN	O	O
patients	NN	O	O
have	NN	O	O
unique	NN	O	O
deletions	NN	O	O
or	NN	O	O
translocations	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
the	NN	O	O
disorder	NN	O	O
in	NN	O	O
the	NN	O	O
population	NN	O	O
and	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
most	NN	O	O
cases	NN	O	O
occur	NN	O	O
sporadically	NN	O	O
suggest	NN	O	O
that	NN	O	O
sequences	NN	O	O
at	NN	O	O
or	NN	O	O
near	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
confer	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
chromosome	NN	O	O
rearrangements	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
hamster	NN	O	O
-	NN	O	O
human	NN	O	O
somatic	NN	O	O
hybrid	NN	O	O
cell	NN	O	O
lines	NN	O	O
from	NN	O	O
VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
all	NN	O	O
three	NN	O	O
classes	NN	O	O
of	NN	O	O
deletions	NN	O	O
and	NN	O	O
we	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
occur	NN	O	O
within	NN	O	O
similar	NN	O	O
low	NN	O	O
copy	NN	O	O
repeats	NN	O	O
,	NN	O	O
termed	NN	O	O
LCR22s	NN	O	O
.	NN	O	O

To	NN	O	O
support	NN	O	O
this	NN	O	O
idea	NN	O	O
further	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
family	NN	O	O
that	NN	O	O
carries	NN	O	O
an	NN	O	O
interstitial	NN	O	O
duplication	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
3	NN	O	O
Mb	NN	O	O
region	NN	O	O
that	NN	O	O
is	NN	O	O
deleted	NN	O	O
in	NN	O	O
VCFS	NN	O	B-Disease
/	NN	O	O
DGS	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
models	NN	O	O
to	NN	O	O
explain	NN	O	O
how	NN	O	O
the	NN	O	O
LCR22s	NN	O	O
can	NN	O	O
mediate	NN	O	O
different	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
events	NN	O	O
,	NN	O	O
thereby	NN	O	O
generating	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
rearrangements	NN	O	O
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
anomaly	NN	O	I-Disease
disorders	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
identified	NN	O	O
five	NN	O	O
additional	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
LCR22	NN	O	O
on	NN	O	O
22q11	NN	O	O
that	NN	O	O
may	NN	O	O
mediate	NN	O	O
other	NN	O	O
rearrangements	NN	O	O
leading	NN	O	O
to	NN	O	O
disease	NN	O	O
.	NN	O	O

Functional	NN	O	O
consequences	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
growth	NN	O	O
response	NN	O	O
2	NN	O	O
gene	NN	O	O
(	NN	O	O
EGR2	NN	O	O
)	NN	O	O
correlate	NN	O	O
with	NN	O	O
severity	NN	O	O
of	NN	O	O
human	NN	O	O
myelinopathies	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
early	NN	O	O
growth	NN	O	O
response	NN	O	O
2	NN	O	O
gene	NN	O	O
(	NN	O	O
EGR2	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
Cys2His2zinc	NN	O	O
finger	NN	O	O
transcription	NN	O	O
factor	NN	O	O
which	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
peripheral	NN	O	O
nervous	NN	O	O
system	NN	O	O
myelination	NN	O	O
.	NN	O	O

This	NN	O	O
idea	NN	O	O
is	NN	O	O
based	NN	O	O
partly	NN	O	O
on	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
homozygous	NN	O	O
Krox20	NN	O	O
(	NN	O	O
Egr2	NN	O	O
)	NN	O	O
knockout	NN	O	O
mice	NN	O	O
,	NN	O	O
which	NN	O	O
display	NN	O	O
hypomyelination	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
PNS	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
block	NN	O	O
of	NN	O	O
Schwann	NN	O	O
cells	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
EGR2	NN	O	O
gene	NN	O	O
have	NN	O	O
recently	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
peripheral	NN	O	I-Disease
neuropathies	NN	O	I-Disease
Charcot	NN	O	B-Disease
-	NN	O	I-Disease
Marie	NN	O	I-Disease
-	NN	O	I-Disease
Tooth	NN	O	I-Disease
type	NN	O	I-Disease
1	NN	O	I-Disease
,	NN	O	O
Dejerine	NN	O	B-Disease
-	NN	O	I-Disease
Sottas	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
congenital	NN	O	B-Disease
hypomyelinating	NN	O	I-Disease
neuropathy	NN	O	I-Disease
.	NN	O	O

Three	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
EGR2	NN	O	O
mutations	NN	O	O
are	NN	O	O
dominant	NN	O	O
and	NN	O	O
occur	NN	O	O
within	NN	O	O
the	NN	O	O
zinc	NN	O	O
finger	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
fourth	NN	O	O
mutation	NN	O	O
is	NN	O	O
recessive	NN	O	O
and	NN	O	O
affects	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
domain	NN	O	O
(	NN	O	O
R1	NN	O	O
)	NN	O	O
that	NN	O	O
binds	NN	O	O
the	NN	O	O
NAB	NN	O	O
transcriptional	NN	O	O
co	NN	O	O
-	NN	O	O
repressors	NN	O	O
.	NN	O	O

A	NN	O	O
combination	NN	O	O
of	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
assays	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
analysis	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
functional	NN	O	O
consequences	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
zinc	NN	O	O
finger	NN	O	O
mutations	NN	O	O
affect	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
residual	NN	O	O
binding	NN	O	O
directly	NN	O	O
correlates	NN	O	O
with	NN	O	O
disease	NN	O	O
severity	NN	O	O
.	NN	O	O

The	NN	O	O
R1	NN	O	O
domain	NN	O	O
mutation	NN	O	O
prevents	NN	O	O
interaction	NN	O	O
of	NN	O	O
EGR2	NN	O	O
with	NN	O	O
the	NN	O	O
NAB	NN	O	O
co	NN	O	O
-	NN	O	O
repressors	NN	O	O
and	NN	O	O
thereby	NN	O	O
increases	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
possible	NN	O	O
disease	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
EGR2	NN	O	O
mutations	NN	O	O
and	NN	O	O
the	NN	O	O
reason	NN	O	O
for	NN	O	O
varying	NN	O	O
severity	NN	O	O
and	NN	O	O
differences	NN	O	O
in	NN	O	O
inheritance	NN	O	O
patterns	NN	O	O
.	NN	O	O
.	NN	O	O

Autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
familial	NN	O	I-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
with	NN	O	O
continued	NN	O	O
secretion	NN	O	O
of	NN	O	O
mutant	NN	O	O
weakly	NN	O	O
active	NN	O	O
vasopressin	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
post	NN	O	O
-	NN	O	O
natal	NN	O	O
development	NN	O	O
of	NN	O	O
arginine	NN	O	B-Disease
vasopressin	NN	O	I-Disease
(	NN	O	I-Disease
AVP	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AVP	NN	O	O
gene	NN	O	O
.	NN	O	O

All	NN	O	O
published	NN	O	O
mutations	NN	O	O
affect	NN	O	O
the	NN	O	O
signal	NN	O	O
peptide	NN	O	O
or	NN	O	O
the	NN	O	O
neurophysin	NN	O	O
-	NN	O	O
II	NN	O	O
carrier	NN	O	O
protein	NN	O	O
and	NN	O	O
are	NN	O	O
presumed	NN	O	O
to	NN	O	O
interfere	NN	O	O
with	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
preprohormone	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
neuronal	NN	O	B-Disease
damage	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
studied	NN	O	O
an	NN	O	O
unusual	NN	O	O
Palestinian	NN	O	O
family	NN	O	O
consisting	NN	O	O
of	NN	O	O
asymptomatic	NN	O	O
first	NN	O	O
cousin	NN	O	O
parents	NN	O	O
and	NN	O	O
three	NN	O	O
children	NN	O	O
affected	NN	O	O
with	NN	O	O
neurohypophyseal	NN	O	B-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
,	NN	O	O
suggesting	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
inheritance	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
affected	NN	O	O
children	NN	O	O
were	NN	O	O
homozygous	NN	O	O
and	NN	O	O
the	NN	O	O
parents	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
single	NN	O	O
novel	NN	O	O
mutation	NN	O	O
(	NN	O	O
C301	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
in	NN	O	O
exon	NN	O	O
1	NN	O	O
,	NN	O	O
replacing	NN	O	O
Pro7	NN	O	O
of	NN	O	O
mature	NN	O	O
AVP	NN	O	O
with	NN	O	O
Leu	NN	O	O
(	NN	O	O
Leu	NN	O	O
-	NN	O	O
AVP	NN	O	O
)	NN	O	O
.	NN	O	O

Leu	NN	O	O
-	NN	O	O
AVP	NN	O	O
was	NN	O	O
a	NN	O	O
weak	NN	O	O
agonist	NN	O	O
with	NN	O	O
approximately	NN	O	O
30	NN	O	O
-	NN	O	O
fold	NN	O	O
reduced	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
V2	NN	O	O
receptor	NN	O	O
.	NN	O	O

Measured	NN	O	O
by	NN	O	O
radioimmunoassay	NN	O	O
with	NN	O	O
a	NN	O	O
synthetic	NN	O	O
Leu	NN	O	O
-	NN	O	O
AVP	NN	O	O
standard	NN	O	O
,	NN	O	O
serum	NN	O	O
Leu	NN	O	O
-	NN	O	O
AVP	NN	O	O
levels	NN	O	O
were	NN	O	O
elevated	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
children	NN	O	O
and	NN	O	O
further	NN	O	O
increased	NN	O	O
during	NN	O	O
water	NN	O	O
deprivation	NN	O	O
to	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
30	NN	O	O
times	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
youngest	NN	O	O
child	NN	O	O
(	NN	O	O
2	NN	O	O
years	NN	O	O
old	NN	O	O
)	NN	O	O
was	NN	O	O
only	NN	O	O
mildly	NN	O	O
affected	NN	O	O
but	NN	O	O
had	NN	O	O
Leu	NN	O	O
-	NN	O	O
AVP	NN	O	O
levels	NN	O	O
similar	NN	O	O
to	NN	O	O
her	NN	O	O
severely	NN	O	O
affected	NN	O	O
8	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
brother	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
unknown	NN	O	O
mechanisms	NN	O	O
may	NN	O	O
partially	NN	O	O
compensate	NN	O	O
for	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
active	NN	O	I-Disease
AVP	NN	O	I-Disease
in	NN	O	O
very	NN	O	O
young	NN	O	O
children	NN	O	O
.	NN	O	O
.	NN	O	O

X	NN	O	O
inactivation	NN	O	O
and	NN	O	O
somatic	NN	O	O
cell	NN	O	O
selection	NN	O	O
rescue	NN	O	O
female	NN	O	O
mice	NN	O	O
carrying	NN	O	O
a	NN	O	O
Piga	NN	O	O
-	NN	O	O
null	NN	O	O
mutation	NN	O	O
.	NN	O	O

A	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
X	NN	O	O
linked	NN	O	O
PIGA	NN	O	O
gene	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
glycosyl	NN	O	I-Disease
phosphatidylinositol	NN	O	I-Disease
(	NN	O	I-Disease
GPI	NN	O	I-Disease
)	NN	O	I-Disease
-	NN	O	I-Disease
anchored	NN	O	I-Disease
proteins	NN	O	I-Disease
on	NN	O	O
blood	NN	O	O
cells	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
paroxysmal	NN	O	B-Disease
nocturnal	NN	O	I-Disease
hemoglobinuria	NN	O	I-Disease
.	NN	O	O

No	NN	O	O
inherited	NN	O	O
form	NN	O	O
of	NN	O	O
GPI	NN	O	B-Disease
-	NN	O	I-Disease
anchor	NN	O	I-Disease
deficiency	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Because	NN	O	O
conventional	NN	O	O
Piga	NN	O	O
gene	NN	O	O
knockout	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
high	NN	O	O
embryonic	NN	O	B-Disease
lethality	NN	O	I-Disease
in	NN	O	O
chimeric	NN	O	O
mice	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
the	NN	O	O
Cre	NN	O	O
/	NN	O	O
loxP	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
mice	NN	O	O
in	NN	O	O
which	NN	O	O
two	NN	O	O
loxP	NN	O	O
sites	NN	O	O
flank	NN	O	O
part	NN	O	O
of	NN	O	O
Piga	NN	O	O
exon	NN	O	O
2	NN	O	O
.	NN	O	O

After	NN	O	O
crossbreeding	NN	O	O
with	NN	O	O
female	NN	O	O
mice	NN	O	O
of	NN	O	O
the	NN	O	O
EIIa	NN	O	O
-	NN	O	O
cre	NN	O	O
strain	NN	O	O
,	NN	O	O
the	NN	O	O
floxed	NN	O	O
allele	NN	O	O
undergoes	NN	O	O
Cre	NN	O	O
-	NN	O	O
mediated	NN	O	O
recombination	NN	O	O
with	NN	O	O
high	NN	O	O
efficiency	NN	O	O
during	NN	O	O
early	NN	O	O
embryonic	NN	O	O
development	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
X	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
,	NN	O	O
female	NN	O	O
offspring	NN	O	O
are	NN	O	O
mosaic	NN	O	O
for	NN	O	O
cells	NN	O	O
that	NN	O	O
express	NN	O	O
or	NN	O	O
lack	NN	O	O
GPI	NN	O	O
-	NN	O	O
linked	NN	O	O
proteins	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
mosaic	NN	O	O
mice	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
heart	NN	O	O
,	NN	O	O
lung	NN	O	O
,	NN	O	O
kidney	NN	O	O
,	NN	O	O
brain	NN	O	O
,	NN	O	O
and	NN	O	O
liver	NN	O	O
,	NN	O	O
mainly	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
Piga	NN	O	O
is	NN	O	O
active	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
tissues	NN	O	O
require	NN	O	O
GPI	NN	O	O
-	NN	O	O
linked	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
salient	NN	O	O
exceptions	NN	O	O
were	NN	O	O
spleen	NN	O	O
,	NN	O	O
thymus	NN	O	O
,	NN	O	O
and	NN	O	O
red	NN	O	O
blood	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
had	NN	O	O
almost	NN	O	O
equal	NN	O	O
numbers	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
or	NN	O	O
the	NN	O	O
recombined	NN	O	O
allele	NN	O	O
,	NN	O	O
implying	NN	O	O
that	NN	O	O
GPI	NN	O	O
-	NN	O	O
linked	NN	O	O
proteins	NN	O	O
are	NN	O	O
not	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
derivation	NN	O	O
of	NN	O	O
these	NN	O	O
tissues	NN	O	O
.	NN	O	O

PIGA	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
cells	NN	O	O
had	NN	O	O
no	NN	O	O
growth	NN	O	O
advantage	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
other	NN	O	O
factors	NN	O	O
are	NN	O	O
needed	NN	O	O
for	NN	O	O
their	NN	O	O
clonal	NN	O	O
dominance	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
paroxysmal	NN	O	B-Disease
nocturnal	NN	O	I-Disease
hemoglobinuria	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

The	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
causing	NN	O	O
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
does	NN	O	O
not	NN	O	O
produce	NN	O	O
a	NN	O	O
null	NN	O	O
allele	NN	O	O
.	NN	O	O

Targeted	NN	O	O
mutagenesis	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
produce	NN	O	O
two	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	O
hemochromatosis	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
Hfe	NN	O	O
)	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
mutation	NN	O	O
deletes	NN	O	O
a	NN	O	O
large	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
,	NN	O	O
generating	NN	O	O
a	NN	O	O
null	NN	O	O
allele	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
mutation	NN	O	O
introduces	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
C282Y	NN	O	O
)	NN	O	O
into	NN	O	O
the	NN	O	O
Hfe	NN	O	O
locus	NN	O	O
,	NN	O	O
but	NN	O	O
otherwise	NN	O	O
leaves	NN	O	O
the	NN	O	O
gene	NN	O	O
intact	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
mutation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
.	NN	O	O

Mice	NN	O	O
carrying	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
were	NN	O	O
bred	NN	O	O
and	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
for	NN	O	O
either	NN	O	O
mutation	NN	O	O
results	NN	O	O
in	NN	O	O
postnatal	NN	O	O
iron	NN	O	O
loading	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
null	NN	O	O
mutation	NN	O	O
are	NN	O	O
more	NN	O	O
severe	NN	O	O
than	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
.	NN	O	O

Mice	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
either	NN	O	O
mutation	NN	O	O
accumulate	NN	O	O
more	NN	O	O
iron	NN	O	O
than	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
although	NN	O	O
liver	NN	O	O
iron	NN	O	O
stores	NN	O	O
are	NN	O	O
greatly	NN	O	O
increased	NN	O	O
,	NN	O	O
splenic	NN	O	O
iron	NN	O	O
is	NN	O	O
decreased	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
null	NN	O	O
allele	NN	O	O
.	NN	O	O
.	NN	O	O

Genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
analysis	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
milder	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
emerin	NN	O	O
gene	NN	O	O
in	NN	O	O
22	NN	O	O
families	NN	O	O
with	NN	O	O
Emery	NN	O	B-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EMD	NN	O	B-Disease
)	NN	O	O
revealed	NN	O	O
mutations	NN	O	O
in	NN	O	O
21	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
emerin	NN	O	O
mutations	NN	O	O
can	NN	O	O
be	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
EMD	NN	O	I-Disease
.	NN	O	O

Most	NN	O	O
emerin	NN	O	O
mutations	NN	O	O
result	NN	O	O
in	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
three	NN	O	O
mutations	NN	O	O
(	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
Pro183Thr	NN	O	O
and	NN	O	O
two	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletions	NN	O	O
removing	NN	O	O
residues	NN	O	O
95	NN	O	O
-	NN	O	O
99	NN	O	O
and	NN	O	O
236	NN	O	O
-	NN	O	O
241	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
were	NN	O	O
unusual	NN	O	O
in	NN	O	O
being	NN	O	O
associated	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
mutant	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
phenotype	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
was	NN	O	O
compared	NN	O	O
in	NN	O	O
detail	NN	O	O
with	NN	O	O
the	NN	O	O
clinical	NN	O	O
features	NN	O	O
in	NN	O	O
cases	NN	O	O
with	NN	O	O
typical	NN	O	O
null	NN	O	O
mutations	NN	O	O
.	NN	O	O

For	NN	O	O
the	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletions	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
significant	NN	O	O
differences	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
family	NN	O	O
with	NN	O	O
the	NN	O	O
missense	NN	O	O
mutation	NN	O	O
the	NN	O	O
phenotype	NN	O	O
was	NN	O	O
milder	NN	O	O
.	NN	O	O

Age	NN	O	O
at	NN	O	O
onset	NN	O	O
was	NN	O	O
later	NN	O	O
for	NN	O	O
first	NN	O	O
symptoms	NN	O	O
and	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
ankle	NN	O	B-Disease
contractures	NN	O	I-Disease
and	NN	O	O
muscle	NN	O	B-Disease
weakness	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
diagnostic	NN	O	O
implications	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
pointing	NN	O	O
to	NN	O	O
functionally	NN	O	O
important	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
emerin	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Severe	NN	O	O
clinical	NN	O	O
expression	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EDMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
relatively	NN	O	O
rare	NN	O	O
benign	NN	O	B-Disease
neuromuscular	NN	O	I-Disease
disorder	NN	O	I-Disease
which	NN	O	O
can	NN	O	O
vary	NN	O	O
remarkably	NN	O	O
in	NN	O	O
onset	NN	O	O
,	NN	O	O
course	NN	O	O
and	NN	O	O
severity	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
a	NN	O	O
TCTAC	NN	O	O
deletion	NN	O	O
spanning	NN	O	O
the	NN	O	O
nucleotides	NN	O	O
631	NN	O	O
-	NN	O	O
635	NN	O	O
of	NN	O	O
the	NN	O	O
emerin	NN	O	O
gene	NN	O	O
caused	NN	O	O
an	NN	O	O
unusually	NN	O	O
severe	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
including	NN	O	O
loss	NN	O	B-Disease
of	NN	O	I-Disease
ambulation	NN	O	I-Disease
and	NN	O	O
severe	NN	O	O
muscle	NN	O	B-Disease
wasting	NN	O	I-Disease
in	NN	O	O
two	NN	O	O
affected	NN	O	O
brothers	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
in	NN	O	O
an	NN	O	O
unrelated	NN	O	O
family	NN	O	O
showing	NN	O	O
a	NN	O	O
significantly	NN	O	O
milder	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
interfamilial	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
distribution	NN	O	O
and	NN	O	O
in	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
features	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
families	NN	O	O
point	NN	O	O
to	NN	O	O
environmental	NN	O	O
or	NN	O	O
genetic	NN	O	O
modification	NN	O	O
as	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
clinical	NN	O	O
variability	NN	O	O
in	NN	O	O
Emery	NN	O	B-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Common	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
BRCA2	NN	O	O
do	NN	O	O
not	NN	O	O
contribute	NN	O	O
to	NN	O	O
early	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
in	NN	O	O
Jewish	NN	O	O
men	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
Families	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
subsequently	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
terminating	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
,	NN	O	O
appear	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
higher	NN	O	O
incidence	NN	O	O
of	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
among	NN	O	O
male	NN	O	O
relatives	NN	O	O
.	NN	O	O

We	NN	O	O
aimed	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
common	NN	O	O
germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
in	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
men	NN	O	O
predisposed	NN	O	O
them	NN	O	O
to	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

METHODS	NN	O	O
We	NN	O	O
examined	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
83	NN	O	O
(	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
185delAG	NN	O	O
)	NN	O	O
or	NN	O	O
82	NN	O	O
(	NN	O	O
for	NN	O	O
BRCA2	NN	O	O
6174delT	NN	O	O
)	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
,	NN	O	O
most	NN	O	O
of	NN	O	O
whom	NN	O	O
were	NN	O	O
treated	NN	O	O
at	NN	O	O
a	NN	O	O
relatively	NN	O	O
young	NN	O	O
age	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
each	NN	O	O
gene	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
population	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Our	NN	O	O
study	NN	O	O
should	NN	O	O
have	NN	O	O
been	NN	O	O
able	NN	O	O
to	NN	O	O
detect	NN	O	O
a	NN	O	O
4	NN	O	O
-	NN	O	O
5	NN	O	O
-	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
mutation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
15	NN	O	O
%	NN	O	O
;	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
,	NN	O	O
0	NN	O	O
-	NN	O	O
3	NN	O	O
.	NN	O	O
6	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
mutant	NN	O	O
allele	NN	O	O
,	NN	O	O
and	NN	O	O
only	NN	O	O
two	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
BRCA2	NN	O	O
mutation	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
4	NN	O	O
%	NN	O	O
;	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
,	NN	O	O
0	NN	O	O
-	NN	O	O
6	NN	O	O
.	NN	O	O
2	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
The	NN	O	O
incidence	NN	O	O
of	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
these	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
closely	NN	O	O
matched	NN	O	O
their	NN	O	O
incidence	NN	O	O
(	NN	O	O
about	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
population	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
unlike	NN	O	O
cases	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
do	NN	O	O
not	NN	O	O
significantly	NN	O	O
predispose	NN	O	O
men	NN	O	O
to	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease

Beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
accumulation	NN	O	O
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
CTNNB1	NN	O	O
gene	NN	O	O
in	NN	O	O
hepatoblastoma	NN	O	B-Disease
.	NN	O	O

Hepatoblastoma	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
malignant	NN	O	B-Disease
tumor	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
liver	NN	O	I-Disease
that	NN	O	O
occurs	NN	O	O
in	NN	O	O
children	NN	O	O
at	NN	O	O
an	NN	O	O
average	NN	O	O
age	NN	O	O
of	NN	O	O
2	NN	O	O
to	NN	O	O
3	NN	O	O
years	NN	O	O
.	NN	O	O

Epidemiologic	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
an	NN	O	O
increased	NN	O	O
frequency	NN	O	O
of	NN	O	O
this	NN	O	O
tumor	NN	O	B-Disease
type	NN	O	O
in	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
epidemiologic	NN	O	O
data	NN	O	O
,	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
tumor	NN	O	I-Disease
suppressor	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
hepatoblastoma	NN	O	B-Disease
tumorigenesis	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
function	NN	O	O
of	NN	O	O
APC	NN	O	O
is	NN	O	O
the	NN	O	O
downregulation	NN	O	O
of	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	O
-	NN	O	O
activating	NN	O	O
protein	NN	O	O
with	NN	O	O
oncogenic	NN	O	O
potential	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
ongoing	NN	O	O
immunohistochemical	NN	O	O
study	NN	O	O
of	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
expression	NN	O	O
in	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
of	NN	O	O
tumor	NN	O	B-Disease
types	NN	O	O
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
observed	NN	O	O
increased	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
three	NN	O	O
investigated	NN	O	O
hepatoblastomas	NN	O	B-Disease
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
exon	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
gene	NN	O	O
(	NN	O	O
CTNNB1	NN	O	O
)	NN	O	O
revealed	NN	O	O
an	NN	O	O
activating	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
samples	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
accumulation	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
hepatoblastoma	NN	O	B-Disease
and	NN	O	O
that	NN	O	O
activating	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
gene	NN	O	O
may	NN	O	O
substitute	NN	O	O
biallelic	NN	O	O
APC	NN	O	O
inactivation	NN	O	O
in	NN	O	O
this	NN	O	O
tumor	NN	O	B-Disease
type	NN	O	O
.	NN	O	O

Genes	NN	O	O
Chromosomes	NN	O	O
Cancer	NN	O	O
25	NN	O	O
399	NN	O	O
-	NN	O	O
402	NN	O	O
,	NN	O	O
1999	NN	O	O
.	NN	O	O
.	NN	O	O

Decrease	NN	O	O
in	NN	O	O
GTP	NN	O	O
cyclohydrolase	NN	O	O
I	NN	O	O
gene	NN	O	O
expression	NN	O	O
caused	NN	O	O
by	NN	O	O
inactivation	NN	O	O
of	NN	O	O
one	NN	O	O
allele	NN	O	O
in	NN	O	O
hereditary	NN	O	B-Disease
progressive	NN	O	I-Disease
dystonia	NN	O	I-Disease
with	NN	O	O
marked	NN	O	O
diurnal	NN	O	O
fluctuation	NN	O	O
.	NN	O	O

Hereditary	NN	O	B-Disease
progressive	NN	O	I-Disease
dystonia	NN	O	I-Disease
with	NN	O	O
marked	NN	O	O
diurnal	NN	O	O
fluctuation	NN	O	O
(	NN	O	O
HPD	NN	O	B-Disease
;	NN	O	O
dopa	NN	O	B-Disease
-	NN	O	I-Disease
responsive	NN	O	I-Disease
dystonia	NN	O	I-Disease
,	NN	O	O
DRD	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
been	NN	O	O
recently	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
GTP	NN	O	O
cyclohydrolase	NN	O	O
I	NN	O	O
(	NN	O	O
GCH1	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
quantified	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
of	NN	O	O
GCH1	NN	O	O
in	NN	O	O
phytohemagglutinin	NN	O	O
(	NN	O	O
PHA	NN	O	O
)	NN	O	O
-	NN	O	O
stimulated	NN	O	O
mononuclear	NN	O	O
blood	NN	O	O
cells	NN	O	O
from	NN	O	O
one	NN	O	O
Japanese	NN	O	O
family	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
or	NN	O	O
splice	NN	O	O
junctions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
GCH1	NN	O	O
mRNA	NN	O	O
were	NN	O	O
decreased	NN	O	O
to	NN	O	O
about	NN	O	O
40	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
level	NN	O	O
in	NN	O	O
both	NN	O	O
patients	NN	O	O
and	NN	O	O
carriers	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
GCH1	NN	O	O
mRNA	NN	O	O
was	NN	O	O
transcribed	NN	O	O
from	NN	O	O
only	NN	O	O
one	NN	O	O
allele	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
other	NN	O	O
allele	NN	O	O
was	NN	O	O
in	NN	O	O
an	NN	O	O
inactive	NN	O	O
state	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
some	NN	O	O
novel	NN	O	O
mutations	NN	O	O
should	NN	O	O
exist	NN	O	O
on	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
alleles	NN	O	O
in	NN	O	O
some	NN	O	O
unknown	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
GCH1	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
decrease	NN	O	O
the	NN	O	O
GCH1	NN	O	O
mRNA	NN	O	O
causing	NN	O	O
the	NN	O	O
HPD	NN	O	B-Disease
/	NN	O	O
DRD	NN	O	B-Disease
symptoms	NN	O	O
.	NN	O	O
.	NN	O	O

Sulfate	NN	O	O
transport	NN	O	O
is	NN	O	O
not	NN	O	O
impaired	NN	O	O
in	NN	O	O
pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
thyrocytes	NN	O	O
.	NN	O	O

Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
syndromic	NN	O	B-Disease
deafness	NN	O	I-Disease
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
dyshormonogenic	NN	O	B-Disease
goiter	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
sensory	NN	O	B-Disease
-	NN	O	I-Disease
neural	NN	O	I-Disease
deafness	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
disease	NN	O	O
(	NN	O	O
PDS	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
,	NN	O	O
but	NN	O	O
its	NN	O	O
function	NN	O	O
is	NN	O	O
as	NN	O	O
yet	NN	O	O
unknown	NN	O	O
and	NN	O	O
the	NN	O	O
connection	NN	O	O
between	NN	O	O
thyroid	NN	O	B-Disease
goiter	NN	O	I-Disease
and	NN	O	O
sensory	NN	O	B-Disease
-	NN	O	I-Disease
neural	NN	O	I-Disease
deafness	NN	O	I-Disease
remains	NN	O	O
an	NN	O	O
enigma	NN	O	O
.	NN	O	O

PDS	NN	O	O
codes	NN	O	O
for	NN	O	O
a	NN	O	O
novel	NN	O	O
protein	NN	O	O
,	NN	O	O
pendrin	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
sufate	NN	O	O
transporters	NN	O	O
.	NN	O	O

Mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
abnormal	NN	O	O
sulfate	NN	O	O
transport	NN	O	O
could	NN	O	O
deleteriously	NN	O	O
affect	NN	O	O
iodide	NN	O	O
organification	NN	O	O
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
.	NN	O	O

We	NN	O	O
tested	NN	O	O
sulfate	NN	O	O
transport	NN	O	O
in	NN	O	O
thyrocytes	NN	O	O
obtained	NN	O	O
from	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
patients	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
not	NN	O	O
defective	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
pendrin	NN	O	O
in	NN	O	O
fact	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
a	NN	O	O
sulfate	NN	O	O
transporter	NN	O	O
,	NN	O	O
and	NN	O	O
emphasizes	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
functional	NN	O	O
studies	NN	O	O
on	NN	O	O
this	NN	O	O
novel	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Small	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
triple	NN	O	O
helix	NN	O	O
produce	NN	O	O
kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
moderately	NN	O	O
severe	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
collagenopathy	NN	O	I-Disease
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
short	NN	O	O
trunk	NN	O	O
and	NN	O	O
limbs	NN	O	O
,	NN	O	O
kyphoscoliosis	NN	O	B-Disease
,	NN	O	O
midface	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
severe	NN	O	O
myopia	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
hearing	NN	O	B-Disease
loss	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
(	NN	O	O
COL2A1	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
predominant	NN	O	O
protein	NN	O	O
of	NN	O	O
cartilage	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

All	NN	O	O
but	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
previously	NN	O	O
described	NN	O	O
mutations	NN	O	O
cause	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletions	NN	O	O
in	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
,	NN	O	O
either	NN	O	O
by	NN	O	O
small	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
or	NN	O	O
splice	NN	O	O
site	NN	O	O
alterations	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
all	NN	O	O
but	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
is	NN	O	O
located	NN	O	O
between	NN	O	O
exons	NN	O	O
12	NN	O	O
and	NN	O	O
24	NN	O	O
in	NN	O	O
the	NN	O	O
COL2A1	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
heteroduplex	NN	O	O
analysis	NN	O	O
to	NN	O	O
identify	NN	O	O
sequence	NN	O	O
anomalies	NN	O	O
in	NN	O	O
five	NN	O	O
individuals	NN	O	O
with	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
index	NN	O	O
patients	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
identified	NN	O	O
four	NN	O	O
new	NN	O	O
dominant	NN	O	O
mutations	NN	O	O
in	NN	O	O
COL2A1	NN	O	O
that	NN	O	O
result	NN	O	O
in	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
a	NN	O	O
21	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
in	NN	O	O
exon	NN	O	O
16	NN	O	O
,	NN	O	O
an	NN	O	O
18	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
in	NN	O	O
exon	NN	O	O
19	NN	O	O
,	NN	O	O
and	NN	O	O
4	NN	O	O
-	NN	O	O
bp	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
splice	NN	O	O
donor	NN	O	O
sites	NN	O	O
of	NN	O	O
introns	NN	O	O
14	NN	O	O
and	NN	O	O
20	NN	O	O
.	NN	O	O

A	NN	O	O
previously	NN	O	O
described	NN	O	O
28	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
at	NN	O	O
the	NN	O	O
COL2A1	NN	O	O
exon	NN	O	O
12	NN	O	O
-	NN	O	O
intron	NN	O	O
12	NN	O	O
junction	NN	O	O
,	NN	O	O
deleting	NN	O	O
the	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
fifth	NN	O	O
case	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
three	NN	O	O
mutations	NN	O	O
are	NN	O	O
predicted	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
exon	NN	O	O
skipping	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
encoded	NN	O	O
from	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
results	NN	O	O
from	NN	O	O
shorter	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
monomers	NN	O	O
,	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
alteration	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
COL2A1	NN	O	O
domain	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
span	NN	O	O
from	NN	O	O
exons	NN	O	O
12	NN	O	O
to	NN	O	O
24	NN	O	O
,	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
phenotype	NN	O	O
.	NN	O	O
.	NN	O	O

Classical	NN	O	B-Disease
galactosemia	NN	O	I-Disease
and	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyl	NN	O	O
transferase	NN	O	O
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Classical	NN	O	B-Disease
galactosemia	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	O
in	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyl	NN	O	O
transferase	NN	O	O
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
GALT	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
disorder	NN	O	O
exhibits	NN	O	O
considerable	NN	O	O
allelic	NN	O	O
heterogeneity	NN	O	O
and	NN	O	O
,	NN	O	O
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
1998	NN	O	O
,	NN	O	O
more	NN	O	O
than	NN	O	O
150	NN	O	O
different	NN	O	O
base	NN	O	O
changes	NN	O	O
were	NN	O	O
recorded	NN	O	O
in	NN	O	O
24	NN	O	O
different	NN	O	O
populations	NN	O	O
and	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
in	NN	O	O
15	NN	O	O
countries	NN	O	O
worldwide	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
most	NN	O	O
frequently	NN	O	O
cited	NN	O	O
are	NN	O	O
Q188R	NN	O	O
,	NN	O	O
K285N	NN	O	O
,	NN	O	O
S135L	NN	O	O
,	NN	O	O
and	NN	O	O
N314D	NN	O	O
.	NN	O	O

Q188R	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
in	NN	O	O
European	NN	O	O
populations	NN	O	O
or	NN	O	O
in	NN	O	O
those	NN	O	O
predominantly	NN	O	O
of	NN	O	O
European	NN	O	O
descent	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
it	NN	O	O
accounts	NN	O	O
for	NN	O	O
60	NN	O	O
-	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
but	NN	O	O
there	NN	O	O
are	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
its	NN	O	O
relative	NN	O	O
frequency	NN	O	O
in	NN	O	O
individual	NN	O	O
populations	NN	O	O
.	NN	O	O

Individuals	NN	O	O
homoallelic	NN	O	O
for	NN	O	O
Q188R	NN	O	O
tend	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
and	NN	O	O
this	NN	O	O
is	NN	O	O
in	NN	O	O
keeping	NN	O	O
with	NN	O	O
the	NN	O	O
virtually	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
observed	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
expression	NN	O	O
systems	NN	O	O
.	NN	O	O

Globally	NN	O	O
,	NN	O	O
K285N	NN	O	O
is	NN	O	O
rarer	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
many	NN	O	O
European	NN	O	O
populations	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
found	NN	O	O
on	NN	O	O
25	NN	O	O
-	NN	O	O
40	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
invariably	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
.	NN	O	O

S135L	NN	O	O
is	NN	O	O
found	NN	O	O
almost	NN	O	O
exclusively	NN	O	O
in	NN	O	O
African	NN	O	O
Americans	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
expression	NN	O	O
results	NN	O	O
are	NN	O	O
discrepant	NN	O	O
,	NN	O	O
but	NN	O	O
some	NN	O	O
individuals	NN	O	O
carrying	NN	O	O
S135L	NN	O	O
appear	NN	O	O
to	NN	O	O
exhibit	NN	O	O
GALT	NN	O	O
activity	NN	O	O
in	NN	O	O
some	NN	O	O
tissues	NN	O	O
.	NN	O	O

Duarte	NN	O	O
1	NN	O	O
(	NN	O	O
or	NN	O	O
Los	NN	O	O
Angeles	NN	O	O
)	NN	O	O
and	NN	O	O
Duarte	NN	O	O
2	NN	O	O
(	NN	O	O
or	NN	O	O
Duarte	NN	O	O
)	NN	O	O
variants	NN	O	O
carry	NN	O	O
the	NN	O	O
same	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
,	NN	O	O
N314D	NN	O	O
,	NN	O	O
even	NN	O	O
though	NN	O	O
D1	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
erythrocyte	NN	O	O
GALT	NN	O	O
activity	NN	O	O
and	NN	O	O
D2	NN	O	O
with	NN	O	O
reduced	NN	O	O
activity	NN	O	O
.	NN	O	O

N314D	NN	O	O
is	NN	O	O
in	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
with	NN	O	O
other	NN	O	O
base	NN	O	O
changes	NN	O	O
that	NN	O	O
differ	NN	O	O
on	NN	O	O
the	NN	O	O
D1	NN	O	O
and	NN	O	O
D2	NN	O	O
alleles	NN	O	O
.	NN	O	O

N314D	NN	O	O
does	NN	O	O
not	NN	O	O
impair	NN	O	O
GALT	NN	O	O
activity	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
expression	NN	O	O
systems	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
GALT	NN	O	O
protein	NN	O	O
in	NN	O	O
lymphoblastoid	NN	O	O
cells	NN	O	O
lines	NN	O	O
from	NN	O	O
D2	NN	O	O
and	NN	O	O
D1	NN	O	O
individuals	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
unclear	NN	O	O
whether	NN	O	O
the	NN	O	O
specific	NN	O	O
molecular	NN	O	O
changes	NN	O	O
that	NN	O	O
distinguish	NN	O	O
the	NN	O	O
D1	NN	O	O
and	NN	O	O
D2	NN	O	O
alleles	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
different	NN	O	O
activities	NN	O	O
.	NN	O	O

The	NN	O	O
considerable	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
documented	NN	O	O
to	NN	O	O
date	NN	O	O
undoubtedly	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
heterogeneity	NN	O	O
that	NN	O	O
is	NN	O	O
observed	NN	O	O
in	NN	O	O
galactosemia	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
additional	NN	O	O
effects	NN	O	O
of	NN	O	O
nonallelic	NN	O	O
variation	NN	O	O
and	NN	O	O
other	NN	O	O
constitutional	NN	O	O
factors	NN	O	O
on	NN	O	O
phenotypic	NN	O	O
variability	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
sporadic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
:	NN	O	O
definition	NN	O	O
of	NN	O	O
a	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
VHL	NN	O	B-Disease
patients	NN	O	O
to	NN	O	O
develop	NN	O	O
an	NN	O	O
RCC	NN	O	B-Disease
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
somatic	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	O
VHL	NN	O	B-Disease
)	NN	O	O
mutations	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
173	NN	O	O
primary	NN	O	O
sporadic	NN	O	O
human	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinomas	NN	O	I-Disease
for	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
tumor	NN	O	I-Disease
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
using	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
and	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

We	NN	O	O
detected	NN	O	O
abnormal	NN	O	O
SSCP	NN	O	O
pattern	NN	O	O
in	NN	O	O
73	NN	O	O
samples	NN	O	O
.	NN	O	O

After	NN	O	O
sequencing	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
microdeletions	NN	O	O
in	NN	O	O
58	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
,	NN	O	O
microinsertions	NN	O	O
in	NN	O	O
17	NN	O	O
%	NN	O	O
,	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
in	NN	O	O
8	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
17	NN	O	O
%	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
mutations	NN	O	O
,	NN	O	O
50	NN	O	O
%	NN	O	O
correspond	NN	O	O
to	NN	O	O
new	NN	O	O
mutations	NN	O	O
.	NN	O	O

VHL	NN	O	B-Disease
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
nonpapillary	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
(	NN	O	O
RCC	NN	O	B-Disease
)	NN	O	O
subtype	NN	O	O
,	NN	O	O
as	NN	O	O
previously	NN	O	O
reported	NN	O	O
.	NN	O	O

To	NN	O	O
compare	NN	O	O
somatic	NN	O	O
and	NN	O	O
germline	NN	O	O
mutations	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
database	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
507	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
of	NN	O	O
mutational	NN	O	O
events	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
difference	NN	O	O
between	NN	O	O
somatic	NN	O	O
and	NN	O	O
germline	NN	O	O
mutations	NN	O	O
with	NN	O	O
mutations	NN	O	O
leading	NN	O	O
to	NN	O	O
truncated	NN	O	O
proteins	NN	O	O
observed	NN	O	O
in	NN	O	O
78	NN	O	O
%	NN	O	O
of	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
vs	NN	O	O
only	NN	O	O
37	NN	O	O
%	NN	O	O
in	NN	O	O
germline	NN	O	O
mutations	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
postulated	NN	O	O
that	NN	O	O
a	NN	O	O
specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
VHL	NN	O	B-Disease
mutations	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
sporadic	NN	O	B-Disease
RCC	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
corresponds	NN	O	O
to	NN	O	O
mutations	NN	O	O
leading	NN	O	O
mainly	NN	O	O
to	NN	O	O
truncated	NN	O	O
proteins	NN	O	O
with	NN	O	O
few	NN	O	O
specific	NN	O	O
missense	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
analyzed	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
RCC	NN	O	B-Disease
in	NN	O	O
VHL	NN	O	B-Disease
families	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
RCC	NN	O	B-Disease
in	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
families	NN	O	O
in	NN	O	O
77	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
with	NN	O	O
mutations	NN	O	O
leading	NN	O	O
to	NN	O	O
truncated	NN	O	O
proteins	NN	O	O
versus	NN	O	O
55	NN	O	O
%	NN	O	O
in	NN	O	O
cases	NN	O	O
with	NN	O	O
missense	NN	O	O
mutations	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
truncated	NN	O	O
proteins	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
higher	NN	O	O
risk	NN	O	O
of	NN	O	O
RCC	NN	O	B-Disease
in	NN	O	O
VHL	NN	O	B-Disease
patients	NN	O	O

Defective	NN	O	O
CTLA	NN	O	O
-	NN	O	O
4	NN	O	O
cycling	NN	O	O
pathway	NN	O	O
in	NN	O	O
Chediak	NN	O	B-Disease
-	NN	O	I-Disease
Higashi	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
for	NN	O	O
deregulation	NN	O	O
of	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

Cytotoxic	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
-	NN	O	O
associated	NN	O	O
antigen	NN	O	O
4	NN	O	O
(	NN	O	O
CTLA	NN	O	O
-	NN	O	O
4	NN	O	O
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
CD152	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Its	NN	O	O
membrane	NN	O	O
expression	NN	O	O
is	NN	O	O
highly	NN	O	O
regulated	NN	O	O
by	NN	O	O
endocytosis	NN	O	O
and	NN	O	O
trafficking	NN	O	O
through	NN	O	O
the	NN	O	O
secretory	NN	O	O
lysosome	NN	O	O
pathway	NN	O	O
.	NN	O	O

Chediak	NN	O	B-Disease
-	NN	O	I-Disease
Higashi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
CHS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
trafficking	NN	O	O
regulator	NN	O	O
gene	NN	O	O
,	NN	O	O
LYST	NN	O	O
.	NN	O	O

It	NN	O	O
results	NN	O	O
in	NN	O	O
defective	NN	O	O
membrane	NN	O	O
targeting	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
present	NN	O	O
in	NN	O	O
secretory	NN	O	O
lysosomes	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
features	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
lymphoproliferative	NN	O	B-Disease
syndrome	NN	O	I-Disease
with	NN	O	O
hemophagocytosis	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
murine	NN	O	O
equivalent	NN	O	O
of	NN	O	O
CHS	NN	O	B-Disease
,	NN	O	O
beige	NN	O	O
mice	NN	O	O
,	NN	O	O
present	NN	O	O
similar	NN	O	O
characteristics	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
develop	NN	O	O
the	NN	O	O
lymphoproliferative	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
show	NN	O	O
herein	NN	O	O
that	NN	O	O
CTLA	NN	O	O
-	NN	O	O
4	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
enlarged	NN	O	O
,	NN	O	O
abnormal	NN	O	O
vesicles	NN	O	O
in	NN	O	O
CHS	NN	O	B-Disease
T	NN	O	O
cells	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
properly	NN	O	O
expressed	NN	O	O
at	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
whereas	NN	O	O
its	NN	O	O
surface	NN	O	O
expression	NN	O	O
is	NN	O	O
not	NN	O	O
impaired	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
therefore	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
defective	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
CTLA	NN	O	O
-	NN	O	O
4	NN	O	O
by	NN	O	O
CHS	NN	O	B-Disease
T	NN	O	O
cells	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
lymphoproliferative	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
observation	NN	O	O
may	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CTLA	NN	O	O
-	NN	O	O
4	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O
.	NN	O	O

Proteolipoprotein	NN	O	O
gene	NN	O	O
analysis	NN	O	O
in	NN	O	O
82	NN	O	O
patients	NN	O	O
with	NN	O	O
sporadic	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
Disease	NN	O	I-Disease
:	NN	O	O
duplications	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
originate	NN	O	O
more	NN	O	O
frequently	NN	O	O
in	NN	O	O
male	NN	O	O
germ	NN	O	O
cells	NN	O	O
,	NN	O	O
but	NN	O	O
point	NN	O	O
mutations	NN	O	O
do	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
Clinical	NN	O	O
European	NN	O	O
Network	NN	O	O
on	NN	O	O
Brain	NN	O	B-Disease
Dysmyelinating	NN	O	I-Disease
Disease	NN	O	I-Disease
.	NN	O	O

Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
Disease	NN	O	I-Disease
(	NN	O	O
PMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
developmental	NN	O	I-Disease
defect	NN	O	I-Disease
of	NN	O	I-Disease
myelination	NN	O	I-Disease
affecting	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
and	NN	O	O
segregating	NN	O	O
with	NN	O	O
the	NN	O	O
proteolipoprotein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
locus	NN	O	O
.	NN	O	O

Investigating	NN	O	O
82	NN	O	O
strictly	NN	O	O
selected	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
of	NN	O	O
PMD	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
found	NN	O	O
PLP	NN	O	O
mutations	NN	O	O
in	NN	O	O
77	NN	O	O
%	NN	O	O
;	NN	O	O
complete	NN	O	O
PLP	NN	O	O
-	NN	O	O
gene	NN	O	O
duplications	NN	O	O
were	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
abnormality	NN	O	O
(	NN	O	O
62	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
coding	NN	O	O
or	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
were	NN	O	O
involved	NN	O	O
less	NN	O	O
frequently	NN	O	O
(	NN	O	O
38	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
maternal	NN	O	O
status	NN	O	O
of	NN	O	O
56	NN	O	O
cases	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
both	NN	O	O
types	NN	O	O
of	NN	O	O
PLP	NN	O	O
mutation	NN	O	O
,	NN	O	O
since	NN	O	O
this	NN	O	O
is	NN	O	O
relevant	NN	O	O
to	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
22	NN	O	O
point	NN	O	O
mutations	NN	O	O
,	NN	O	O
68	NN	O	O
%	NN	O	O
of	NN	O	O
mothers	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
value	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
two	NN	O	O
-	NN	O	O
thirds	NN	O	O
of	NN	O	O
carrier	NN	O	O
mothers	NN	O	O
that	NN	O	O
would	NN	O	O
be	NN	O	O
expected	NN	O	O
if	NN	O	O
there	NN	O	O
were	NN	O	O
an	NN	O	O
equal	NN	O	O
mutation	NN	O	O
rate	NN	O	O
in	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
germ	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
sharp	NN	O	O
contrast	NN	O	O
,	NN	O	O
among	NN	O	O
the	NN	O	O
34	NN	O	O
duplicated	NN	O	O
cases	NN	O	O
,	NN	O	O
91	NN	O	O
%	NN	O	O
of	NN	O	O
mothers	NN	O	O
were	NN	O	O
carriers	NN	O	O
,	NN	O	O
a	NN	O	O
value	NN	O	O
significantly	NN	O	O
(	NN	O	O
chi2	NN	O	O
=	NN	O	O
9	NN	O	O
.	NN	O	O
20	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
.	NN	O	O
01	NN	O	O
)	NN	O	O
in	NN	O	O
favor	NN	O	O
of	NN	O	O
a	NN	O	O
male	NN	O	O
bias	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
estimation	NN	O	O
of	NN	O	O
the	NN	O	O
male	NN	O	O
/	NN	O	O
female	NN	O	O
mutation	NN	O	O
frequency	NN	O	O
(	NN	O	O
k	NN	O	O
)	NN	O	O
of	NN	O	O
9	NN	O	O
.	NN	O	O

3	NN	O	O
3	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
mutations	NN	O	O
between	NN	O	O
parental	NN	O	O
and	NN	O	O
grandparental	NN	O	O
generations	NN	O	O
in	NN	O	O
17	NN	O	O
three	NN	O	O
-	NN	O	O
generation	NN	O	O
families	NN	O	O
,	NN	O	O
which	NN	O	O
allowed	NN	O	O
a	NN	O	O
direct	NN	O	O
estimation	NN	O	O
of	NN	O	O
the	NN	O	O
k	NN	O	O
value	NN	O	O
(	NN	O	O
k	NN	O	O
=	NN	O	O
11	NN	O	O
)	NN	O	O
.	NN	O	O

Again	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
male	NN	O	O
mutation	NN	O	O
imbalance	NN	O	O
was	NN	O	O
observed	NN	O	O
only	NN	O	O
for	NN	O	O
the	NN	O	O
duplications	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
breakage	NN	O	O
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
and	NN	O	I-Disease
Angelman	NN	O	I-Disease
syndromes	NN	O	I-Disease
involves	NN	O	O
recombination	NN	O	O
between	NN	O	O
large	NN	O	O
,	NN	O	O
transcribed	NN	O	O
repeats	NN	O	O
at	NN	O	O
proximal	NN	O	O
and	NN	O	O
distal	NN	O	O
breakpoints	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
distinct	NN	O	O
neurobehavioral	NN	O	B-Disease
disorders	NN	O	I-Disease
that	NN	O	O
most	NN	O	O
often	NN	O	O
arise	NN	O	O
from	NN	O	O
a	NN	O	O
4	NN	O	O
-	NN	O	O
Mb	NN	O	O
deletion	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
during	NN	O	O
paternal	NN	O	O
or	NN	O	O
maternal	NN	O	O
gametogenesis	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

At	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
frequency	NN	O	O
of	NN	O	O
approximately	NN	O	O
.	NN	O	O

67	NN	O	O
-	NN	O	O
1	NN	O	O
/	NN	O	O
10	NN	O	O
,	NN	O	O
000	NN	O	O
births	NN	O	O
,	NN	O	O
these	NN	O	O
deletions	NN	O	O
represent	NN	O	O
a	NN	O	O
common	NN	O	O
structural	NN	O	O
chromosome	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
genome	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
these	NN	O	O
events	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
the	NN	O	O
regions	NN	O	O
that	NN	O	O
contain	NN	O	O
two	NN	O	O
proximal	NN	O	O
breakpoint	NN	O	O
clusters	NN	O	O
and	NN	O	O
a	NN	O	O
distal	NN	O	O
cluster	NN	O	O
.	NN	O	O

Novel	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
potentially	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
were	NN	O	O
positionally	NN	O	O
cloned	NN	O	O
from	NN	O	O
YACs	NN	O	O
within	NN	O	O
or	NN	O	O
near	NN	O	O
these	NN	O	O
regions	NN	O	O
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
rodent	NN	O	O
-	NN	O	O
human	NN	O	O
somatic	NN	O	O
-	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
,	NN	O	O
YAC	NN	O	O
contigs	NN	O	O
,	NN	O	O
and	NN	O	O
FISH	NN	O	O
of	NN	O	O
normal	NN	O	O
or	NN	O	O
rearranged	NN	O	O
chromosomes	NN	O	O
15	NN	O	O
identified	NN	O	O
duplicated	NN	O	O
sequences	NN	O	O
(	NN	O	O
the	NN	O	O
END	NN	O	O
repeats	NN	O	O
)	NN	O	O
at	NN	O	O
or	NN	O	O
near	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
.	NN	O	O

The	NN	O	O
END	NN	O	O
-	NN	O	O
repeat	NN	O	O
units	NN	O	O
are	NN	O	O
derived	NN	O	O
from	NN	O	O
large	NN	O	O
genomic	NN	O	O
duplications	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
(	NN	O	O
HERC2	NN	O	O
)	NN	O	O
,	NN	O	O
many	NN	O	O
copies	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
in	NN	O	O
germline	NN	O	O
tissues	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
five	NN	O	O
PWS	NN	O	B-Disease
/	NN	O	O
AS	NN	O	B-Disease
patients	NN	O	O
analyzed	NN	O	O
to	NN	O	O
date	NN	O	O
has	NN	O	O
an	NN	O	O
identifiable	NN	O	O
,	NN	O	O
rearranged	NN	O	O
HERC2	NN	O	O
transcript	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
deletion	NN	O	O
event	NN	O	O
.	NN	O	O

We	NN	O	O
postulate	NN	O	O
that	NN	O	O
the	NN	O	O
END	NN	O	O
repeats	NN	O	O
flanking	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
mediate	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
resulting	NN	O	O
in	NN	O	O
deletion	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
active	NN	O	O
transcription	NN	O	O
of	NN	O	O
these	NN	O	O
repeats	NN	O	O
in	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
germ	NN	O	O
cells	NN	O	O
may	NN	O	O
facilitate	NN	O	O
the	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
process	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
Brazilian	NN	O	O
family	NN	O	O
with	NN	O	O
van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
suggests	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
susceptibility	NN	O	O
locus	NN	O	O
for	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
at	NN	O	O
17p11	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
11	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O

van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
VWS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
1q32	NN	O	O
-	NN	O	O
41	NN	O	O
,	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
pits	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
sinuses	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
lower	NN	O	I-Disease
lip	NN	O	I-Disease
,	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
/	NN	O	I-Disease
palate	NN	O	I-Disease
(	NN	O	O
CL	NN	O	B-Disease
/	NN	O	I-Disease
P	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
(	NN	O	O
CP	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
bifid	NN	O	B-Disease
uvula	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hypodontia	NN	O	B-Disease
(	NN	O	O
H	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
VWS	NN	O	B-Disease
,	NN	O	O
which	NN	O	O
has	NN	O	O
incomplete	NN	O	O
penetrance	NN	O	O
,	NN	O	O
is	NN	O	O
highly	NN	O	O
variable	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
CL	NN	O	B-Disease
/	NN	O	I-Disease
P	NN	O	I-Disease
and	NN	O	O
CP	NN	O	B-Disease
within	NN	O	O
the	NN	O	O
same	NN	O	O
genealogy	NN	O	O
and	NN	O	O
a	NN	O	O
recurrence	NN	O	O
risk	NN	O	O
<	NN	O	O
40	NN	O	O
%	NN	O	O
for	NN	O	O
CP	NN	O	B-Disease
among	NN	O	O
descendants	NN	O	O
with	NN	O	O
VWS	NN	O	B-Disease
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
clefts	NN	O	B-Disease
in	NN	O	O
this	NN	O	O
syndrome	NN	O	O
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
modifying	NN	O	O
genes	NN	O	O
at	NN	O	O
other	NN	O	O
loci	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
conducted	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
Brazilian	NN	O	O
kindred	NN	O	O
with	NN	O	O
VWS	NN	O	B-Disease
,	NN	O	O
considering	NN	O	O
as	NN	O	O
affected	NN	O	O
the	NN	O	O
individuals	NN	O	O
with	NN	O	O
CP	NN	O	B-Disease
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
whether	NN	O	O
it	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
other	NN	O	O
clinical	NN	O	O
signs	NN	O	O
of	NN	O	O
VWS	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
gene	NN	O	O
at	NN	O	O
17p11	NN	O	O
.	NN	O	O

2	NN	O	O
-	NN	O	O
11	NN	O	O
2	NN	O	O
-	NN	O	O
11	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
at	NN	O	O
1p32	NN	O	O
-	NN	O	O
41	NN	O	O
,	NN	O	O
enhances	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
CP	NN	O	B-Disease
in	NN	O	O
an	NN	O	O
individual	NN	O	O
carrying	NN	O	O
the	NN	O	O
two	NN	O	O
at	NN	O	O
-	NN	O	O
risk	NN	O	O
genes	NN	O	O
.	NN	O	O

If	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
is	NN	O	O
confirmed	NN	O	O
in	NN	O	O
other	NN	O	O
VWS	NN	O	B-Disease
pedigrees	NN	O	O
,	NN	O	O
it	NN	O	O
will	NN	O	O
represent	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
examples	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
,	NN	O	O
mapped	NN	O	O
through	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
,	NN	O	O
which	NN	O	O
modifies	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
major	NN	O	O
gene	NN	O	O
.	NN	O	O

New	NN	O	O
mutations	NN	O	O
,	NN	O	O
polymorphisms	NN	O	O
,	NN	O	O
and	NN	O	O
rare	NN	O	O
variants	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
detected	NN	O	O
by	NN	O	O
a	NN	O	O
novel	NN	O	O
SSCP	NN	O	O
strategy	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
,	NN	O	O
ATM	NN	O	O
,	NN	O	O
spans	NN	O	O
about	NN	O	O
150	NN	O	O
kb	NN	O	O
of	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
.	NN	O	O

ATM	NN	O	O
mutations	NN	O	O
are	NN	O	O
found	NN	O	O
along	NN	O	O
the	NN	O	O
entire	NN	O	O
gene	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
mutational	NN	O	O
hot	NN	O	O
spot	NN	O	O
.	NN	O	O

Using	NN	O	O
DNA	NN	O	O
as	NN	O	O
the	NN	O	O
starting	NN	O	O
material	NN	O	O
,	NN	O	O
we	NN	O	O
screened	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
in	NN	O	O
92	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
,	NN	O	O
using	NN	O	O
an	NN	O	O
optimized	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
technique	NN	O	O
that	NN	O	O
detected	NN	O	O
all	NN	O	O
previously	NN	O	O
known	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
segments	NN	O	O
being	NN	O	O
analyzed	NN	O	O
.	NN	O	O

To	NN	O	O
expedite	NN	O	O
screening	NN	O	O
,	NN	O	O
we	NN	O	O
sequentially	NN	O	O
loaded	NN	O	O
the	NN	O	O
SSCP	NN	O	O
gels	NN	O	O
with	NN	O	O
three	NN	O	O
different	NN	O	O
sets	NN	O	O
of	NN	O	O
PCR	NN	O	O
products	NN	O	O
that	NN	O	O
were	NN	O	O
pretested	NN	O	O
to	NN	O	O
avoid	NN	O	O
overlapping	NN	O	O
patterns	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
changes	NN	O	O
we	NN	O	O
detected	NN	O	O
were	NN	O	O
intragenic	NN	O	O
polymorphisms	NN	O	O
.	NN	O	O

Of	NN	O	O
an	NN	O	O
expected	NN	O	O
177	NN	O	O
unknown	NN	O	O
mutations	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
approximately	NN	O	O
70	NN	O	O
%	NN	O	O
,	NN	O	O
mostly	NN	O	O
protein	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
(	NN	O	O
that	NN	O	O
would	NN	O	O
have	NN	O	O
been	NN	O	O
detectable	NN	O	O
by	NN	O	O
protein	NN	O	O
truncation	NN	O	O
testing	NN	O	O
if	NN	O	O
RNA	NN	O	O
starting	NN	O	O
material	NN	O	O
had	NN	O	O
been	NN	O	O
available	NN	O	O
)	NN	O	O
.	NN	O	O

Mutations	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
defined	NN	O	O
for	NN	O	O
every	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
.	NN	O	O

Herein	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
35	NN	O	O
new	NN	O	O
mutations	NN	O	O
and	NN	O	O
34	NN	O	O
new	NN	O	O
intragenic	NN	O	O
polymorphisms	NN	O	O
or	NN	O	O
rare	NN	O	O
variants	NN	O	O
within	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
comprehensive	NN	O	O
compilation	NN	O	O
of	NN	O	O
ATM	NN	O	O
polymorphisms	NN	O	O
assembled	NN	O	O
to	NN	O	O
date	NN	O	O
.	NN	O	O

Defining	NN	O	O
polymorphic	NN	O	O
sites	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
will	NN	O	O
be	NN	O	O
of	NN	O	O
great	NN	O	O
importance	NN	O	O
in	NN	O	O
designing	NN	O	O
automated	NN	O	O
methods	NN	O	O
for	NN	O	O
detecting	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
McLeod	NN	O	B-Disease
syndrome	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
family	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
XK	NN	O	O
gene	NN	O	O
(	NN	O	O
XK	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
patient	NN	O	O
with	NN	O	O
McLeod	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
50	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
man	NN	O	O
showed	NN	O	O
progressive	NN	O	O
muscular	NN	O	B-Disease
atrophy	NN	O	I-Disease
,	NN	O	O
choreic	NN	O	B-Disease
movement	NN	O	I-Disease
,	NN	O	O
elevated	NN	O	O
level	NN	O	O
of	NN	O	O
serum	NN	O	O
creatinine	NN	O	O
kinase	NN	O	O
,	NN	O	O
and	NN	O	O
acanthocytosis	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
Kell	NN	O	O
antigens	NN	O	O
in	NN	O	O
erythrocyte	NN	O	O
was	NN	O	O
decreased	NN	O	O
and	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
-	NN	O	O
base	NN	O	O
(	NN	O	O
T	NN	O	O
)	NN	O	O
deletion	NN	O	O
at	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
1095	NN	O	O
in	NN	O	O
XK	NN	O	O
.	NN	O	O

This	NN	O	O
deletion	NN	O	O
caused	NN	O	O
a	NN	O	O
frameshift	NN	O	O
in	NN	O	O
translation	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
at	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
position	NN	O	O
408	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
this	NN	O	O
single	NN	O	O
-	NN	O	O
base	NN	O	O
deletion	NN	O	O
causes	NN	O	O
defective	NN	O	O
Kx	NN	O	O
protein	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
McLeod	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O
.	NN	O	O

Association	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
with	NN	O	O
the	NN	O	O
hRad50	NN	O	O
-	NN	O	O
hMre11	NN	O	O
-	NN	O	O
p95	NN	O	O
complex	NN	O	O
and	NN	O	O
the	NN	O	O
DNA	NN	O	O
damage	NN	O	O
response	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
encodes	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
that	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

Here	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
interacts	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
hRad50	NN	O	O
,	NN	O	O
which	NN	O	O
forms	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
hMre11	NN	O	O
and	NN	O	O
p95	NN	O	O
/	NN	O	O
nibrin	NN	O	O
.	NN	O	O

Upon	NN	O	O
irradiation	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
discrete	NN	O	O
foci	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
which	NN	O	O
colocalize	NN	O	O
with	NN	O	O
hRad50	NN	O	O
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
irradiation	NN	O	O
-	NN	O	O
induced	NN	O	O
foci	NN	O	O
positive	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
hRad50	NN	O	O
,	NN	O	O
hMre11	NN	O	O
,	NN	O	O
or	NN	O	O
p95	NN	O	O
was	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
in	NN	O	O
HCC	NN	O	O
/	NN	O	O
1937	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
cells	NN	O	O
carrying	NN	O	O
a	NN	O	O
homozygous	NN	O	O
mutation	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
but	NN	O	O
was	NN	O	O
restored	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mutated	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
rendered	NN	O	O
them	NN	O	O
less	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
damage	NN	O	O
agent	NN	O	O
,	NN	O	O
methyl	NN	O	O
methanesulfonate	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
cellular	NN	O	O
responses	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage	NN	O	O
that	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
hRad50	NN	O	O
-	NN	O	O
hMre11	NN	O	O
-	NN	O	O
p95	NN	O	O
complex	NN	O	O
.	NN	O	O
.	NN	O	O

Relationship	NN	O	O
among	NN	O	O
genotype	NN	O	O
,	NN	O	O
biochemical	NN	O	O
phenotype	NN	O	O
,	NN	O	O
and	NN	O	O
cognitive	NN	O	O
performance	NN	O	O
in	NN	O	O
females	NN	O	O
with	NN	O	O
phenylalanine	NN	O	B-Disease
hydroxylase	NN	O	I-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
report	NN	O	O
from	NN	O	O
the	NN	O	O
Maternal	NN	O	B-Disease
Phenylketonuria	NN	O	I-Disease
Collaborative	NN	O	O
Study	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
To	NN	O	O
examine	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
genotypes	NN	O	O
to	NN	O	O
biochemical	NN	O	O
phenotype	NN	O	O
and	NN	O	O
cognitive	NN	O	O
development	NN	O	O
in	NN	O	O
maternal	NN	O	B-Disease
phenylketonuria	NN	O	I-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

METHODOLOGY	NN	O	O
PAH	NN	O	O
gene	NN	O	O
mutations	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
222	NN	O	O
hyperphenylalaninemic	NN	O	B-Disease
females	NN	O	O
enrolled	NN	O	O
in	NN	O	O
the	NN	O	O
Maternal	NN	O	B-Disease
PKU	NN	O	I-Disease
Collaborative	NN	O	O
Study	NN	O	O
(	NN	O	O
MPKUCS	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
84	NN	O	O
different	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
,	NN	O	O
and	NN	O	O
complete	NN	O	O
genotype	NN	O	O
was	NN	O	O
obtained	NN	O	O
in	NN	O	O
199	NN	O	O
individuals	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
previous	NN	O	O
knowledge	NN	O	O
about	NN	O	O
mutation	NN	O	O
-	NN	O	O
phenotype	NN	O	O
associations	NN	O	O
,	NN	O	O
78	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
could	NN	O	O
be	NN	O	O
assigned	NN	O	O
to	NN	O	O
one	NN	O	O
of	NN	O	O
four	NN	O	O
classes	NN	O	O
of	NN	O	O
severity	NN	O	O
(	NN	O	O
severe	NN	O	O
PKU	NN	O	B-Disease
,	NN	O	O
moderate	NN	O	O
PKU	NN	O	B-Disease
,	NN	O	O
mild	NN	O	O
PKU	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
mild	NN	O	B-Disease
hyperphenylalaninemia	NN	O	I-Disease
[	NN	O	O
MHP	NN	O	B-Disease
]	NN	O	O
)	NN	O	O
.	NN	O	O

Then	NN	O	O
,	NN	O	O
189	NN	O	O
MPKUCS	NN	O	O
subjects	NN	O	O
were	NN	O	O
grouped	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
various	NN	O	O
combinations	NN	O	O
of	NN	O	O
mutation	NN	O	O
classifications	NN	O	O
.	NN	O	O

The	NN	O	O
sample	NN	O	O
sizes	NN	O	O
were	NN	O	O
large	NN	O	O
enough	NN	O	O
for	NN	O	O
statistical	NN	O	O
testing	NN	O	O
in	NN	O	O
four	NN	O	O
groups	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mutation	NN	O	O
that	NN	O	O
completely	NN	O	O
abolishes	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
patients	NN	O	O
are	NN	O	O
considered	NN	O	O
functionally	NN	O	O
hemizygous	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
The	NN	O	O
biochemical	NN	O	O
phenotype	NN	O	O
predicted	NN	O	O
from	NN	O	O
the	NN	O	O
genotype	NN	O	O
in	NN	O	O
functionally	NN	O	O
hemizygous	NN	O	O
patients	NN	O	O
was	NN	O	O
related	NN	O	O
significantly	NN	O	O
to	NN	O	O
the	NN	O	O
assigned	NN	O	O
phenylalanine	NN	O	O
level	NN	O	O
.	NN	O	O

Cognitive	NN	O	O
performance	NN	O	O
(	NN	O	O
IQ	NN	O	O
)	NN	O	O
was	NN	O	O
also	NN	O	O
significantly	NN	O	O
related	NN	O	O
to	NN	O	O
genotype	NN	O	O
.	NN	O	O

The	NN	O	O
IQ	NN	O	O
of	NN	O	O
PAH	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
mothers	NN	O	O
with	NN	O	O
a	NN	O	O
severe	NN	O	O
PKU	NN	O	B-Disease
mutation	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
a	NN	O	O
MHP	NN	O	B-Disease
mutation	NN	O	O
or	NN	O	O
a	NN	O	O
mild	NN	O	O
PKU	NN	O	B-Disease
mutation	NN	O	O
was	NN	O	O
99	NN	O	O
and	NN	O	O
96	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
IQ	NN	O	O
of	NN	O	O
PKU	NN	O	B-Disease
mothers	NN	O	O
with	NN	O	O
two	NN	O	O
severe	NN	O	O
PKU	NN	O	B-Disease
mutations	NN	O	O
or	NN	O	O
with	NN	O	O
one	NN	O	O
severe	NN	O	O
and	NN	O	O
one	NN	O	O
moderate	NN	O	O
PKU	NN	O	B-Disease
mutation	NN	O	O
was	NN	O	O
83	NN	O	O
and	NN	O	O
84	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
PKU	NN	O	B-Disease
,	NN	O	O
92	NN	O	O
%	NN	O	O
had	NN	O	O
been	NN	O	O
treated	NN	O	O
during	NN	O	O
childhood	NN	O	O
.	NN	O	O

Those	NN	O	O
who	NN	O	O
were	NN	O	O
untreated	NN	O	O
or	NN	O	O
treated	NN	O	O
late	NN	O	O
had	NN	O	O
lower	NN	O	O
than	NN	O	O
average	NN	O	O
IQ	NN	O	O
scores	NN	O	O
for	NN	O	O
their	NN	O	O
group	NN	O	O
of	NN	O	O
mutation	NN	O	O
combinations	NN	O	O
.	NN	O	O

Females	NN	O	O
with	NN	O	O
moderate	NN	O	O
or	NN	O	O
mild	NN	O	O
PKU	NN	O	B-Disease
who	NN	O	O
were	NN	O	O
treated	NN	O	O
early	NN	O	O
and	NN	O	O
treated	NN	O	O
for	NN	O	O
>	NN	O	O
6	NN	O	O
years	NN	O	O
showed	NN	O	O
IQ	NN	O	O
scores	NN	O	O
10	NN	O	O
points	NN	O	O
above	NN	O	O
average	NN	O	O
for	NN	O	O
their	NN	O	O
group	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
The	NN	O	O
reproductive	NN	O	O
outcome	NN	O	O
in	NN	O	O
maternal	NN	O	B-Disease
phenylketonuria	NN	O	I-Disease
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
prenatal	NN	O	O
metabolic	NN	O	O
control	NN	O	O
and	NN	O	O
postnatal	NN	O	O
environmental	NN	O	O
circumstances	NN	O	O
.	NN	O	O

Both	NN	O	O
factors	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
intellectual	NN	O	O
resources	NN	O	O
of	NN	O	O
the	NN	O	O
mother	NN	O	O
with	NN	O	O
PKU	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
significant	NN	O	O
relationship	NN	O	O
among	NN	O	O
genotype	NN	O	O
,	NN	O	O
biochemical	NN	O	O
phenotype	NN	O	O
,	NN	O	O
and	NN	O	O
cognitive	NN	O	O
performance	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
is	NN	O	O
of	NN	O	O
importance	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
optimal	NN	O	O
strategy	NN	O	O
for	NN	O	O
future	NN	O	O
treatment	NN	O	O
of	NN	O	O
females	NN	O	O
with	NN	O	O
PKU	NN	O	B-Disease
who	NN	O	O
plan	NN	O	O
pregnancy	NN	O	O
.	NN	O	O
.	NN	O	O

Spinal	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
variant	NN	O	O
of	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
describe	NN	O	O
seven	NN	O	O
Dutch	NN	O	O
patients	NN	O	O
from	NN	O	O
six	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
slowly	NN	O	O
progressive	NN	O	O
,	NN	O	O
mainly	NN	O	O
spinal	NN	O	B-Disease
cord	NN	O	I-Disease
syndrome	NN	O	I-Disease
that	NN	O	O
remained	NN	O	O
for	NN	O	O
many	NN	O	O
years	NN	O	O
the	NN	O	O
sole	NN	O	O
expression	NN	O	O
of	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

MRI	NN	O	O
demonstrated	NN	O	O
white	NN	O	B-Disease
matter	NN	O	I-Disease
abnormalities	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
lateral	NN	O	O
and	NN	O	O
dorsal	NN	O	O
columns	NN	O	O
of	NN	O	O
the	NN	O	O
spinal	NN	O	O
cord	NN	O	O
.	NN	O	O

Post	NN	O	O
-	NN	O	O
mortem	NN	O	O
examination	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
showed	NN	O	O
extensive	NN	O	O
myelin	NN	O	O
loss	NN	O	O
in	NN	O	O
these	NN	O	O
columns	NN	O	O
.	NN	O	O

An	NN	O	O
array	NN	O	O
of	NN	O	O
genotypes	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
spinal	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
clinical	NN	O	O
and	NN	O	O
radiological	NN	O	O
separate	NN	O	O
entity	NN	O	O
of	NN	O	O
CTX	NN	O	B-Disease
that	NN	O	O
should	NN	O	O
be	NN	O	O
included	NN	O	O
in	NN	O	O
the	NN	O	O
differential	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
chronic	NN	O	B-Disease
myelopathy	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
transgene	NN	O	O
insertion	NN	O	O
creating	NN	O	O
a	NN	O	O
heritable	NN	O	O
chromosome	NN	O	O
deletion	NN	O	O
mouse	NN	O	O
model	NN	O	O
of	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
and	NN	O	I-Disease
angelman	NN	O	I-Disease
syndromes	NN	O	I-Disease
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
result	NN	O	O
from	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
function	NN	O	O
of	NN	O	O
imprinted	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
.	NN	O	O

The	NN	O	O
central	NN	O	O
part	NN	O	O
of	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
7	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
human	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
,	NN	O	O
with	NN	O	O
conservation	NN	O	O
of	NN	O	O
both	NN	O	O
gene	NN	O	O
order	NN	O	O
and	NN	O	O
imprinted	NN	O	O
features	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
transgene	NN	O	O
insertion	NN	O	O
(	NN	O	O
Epstein	NN	O	O
-	NN	O	O
Barr	NN	O	O
virus	NN	O	O
Latent	NN	O	O
Membrane	NN	O	O
Protein	NN	O	O
2A	NN	O	O
,	NN	O	O
LMP2A	NN	O	O
)	NN	O	O
into	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
7C	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
resulted	NN	O	O
in	NN	O	O
mouse	NN	O	O
models	NN	O	O
for	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
AS	NN	O	B-Disease
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
sex	NN	O	O
of	NN	O	O
the	NN	O	O
transmitting	NN	O	O
parent	NN	O	O
.	NN	O	O

Epigenotype	NN	O	O
(	NN	O	O
allelic	NN	O	O
expression	NN	O	O
and	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
)	NN	O	O
and	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
transgene	NN	O	O
-	NN	O	O
induced	NN	O	O
mutation	NN	O	O
has	NN	O	O
generated	NN	O	O
a	NN	O	O
complete	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
/	NN	O	O
AS	NN	O	B-Disease
-	NN	O	O
homologous	NN	O	O
region	NN	O	O
but	NN	O	O
has	NN	O	O
not	NN	O	O
deleted	NN	O	O
flanking	NN	O	O
loci	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
intact	NN	O	O
chromosome	NN	O	O
7	NN	O	O
,	NN	O	O
opposite	NN	O	O
the	NN	O	O
deleted	NN	O	O
homolog	NN	O	O
,	NN	O	O
maintains	NN	O	O
the	NN	O	O
correct	NN	O	O
imprint	NN	O	O
in	NN	O	O
somatic	NN	O	O
cells	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
AS	NN	O	B-Disease
mice	NN	O	O
and	NN	O	O
establishes	NN	O	O
the	NN	O	O
correct	NN	O	O
imprint	NN	O	O
in	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
germ	NN	O	O
cells	NN	O	O
of	NN	O	O
AS	NN	O	B-Disease
mice	NN	O	O
,	NN	O	O
homologous	NN	O	O
association	NN	O	O
and	NN	O	O
replication	NN	O	O
asynchrony	NN	O	O
are	NN	O	O
not	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
imprinting	NN	O	O
mechanism	NN	O	O
.	NN	O	O

This	NN	O	O
heritable	NN	O	O
-	NN	O	O
deletion	NN	O	O
mouse	NN	O	O
model	NN	O	O
will	NN	O	O
be	NN	O	O
particularly	NN	O	O
useful	NN	O	O
for	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
etiological	NN	O	O
genes	NN	O	O
and	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
phenotypic	NN	O	O
basis	NN	O	O
,	NN	O	O
and	NN	O	O
investigation	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
approaches	NN	O	O
for	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
5	NN	O	O
novel	NN	O	O
van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
kindreds	NN	O	O
to	NN	O	O
1q32	NN	O	O
-	NN	O	O
q41	NN	O	O
markers	NN	O	O
further	NN	O	O
supports	NN	O	O
locus	NN	O	O
homogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
trait	NN	O	O
.	NN	O	O

van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
vWS	NN	O	B-Disease
,	NN	O	O
MIM	NN	O	O
119300	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
clefting	NN	O	I-Disease
condition	NN	O	I-Disease
with	NN	O	O
cardinal	NN	O	O
features	NN	O	O
of	NN	O	O
mucous	NN	O	B-Disease
cysts	NN	O	I-Disease
(	NN	O	O
lower	NN	O	B-Disease
-	NN	O	I-Disease
lip	NN	O	I-Disease
pits	NN	O	I-Disease
)	NN	O	O
and	NN	O	O
clefts	NN	O	B-Disease
to	NN	O	I-Disease
the	NN	O	I-Disease
lip	NN	O	I-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
palate	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
vWS	NN	O	B-Disease
gene	NN	O	O
has	NN	O	O
been	NN	O	O
assigned	NN	O	O
to	NN	O	O
a	NN	O	O
locus	NN	O	O
in	NN	O	O
1q32	NN	O	O
-	NN	O	O
q41	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
and	NN	O	O
physical	NN	O	O
mapping	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
5	NN	O	O
novel	NN	O	O
vWS	NN	O	B-Disease
families	NN	O	O
through	NN	O	O
probands	NN	O	O
attended	NN	O	O
for	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
palate	NN	O	I-Disease
repair	NN	O	O
at	NN	O	O
the	NN	O	O
Department	NN	O	O
of	NN	O	O
Maxillofacial	NN	O	O
Surgery	NN	O	O
of	NN	O	O
Hopital	NN	O	O
Trousseau	NN	O	O
,	NN	O	O
Paris	NN	O	O
,	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
tentatively	NN	O	O
refine	NN	O	O
the	NN	O	O
genetic	NN	O	O
map	NN	O	O
of	NN	O	O
the	NN	O	O
vWS	NN	O	B-Disease
region	NN	O	O
in	NN	O	O
1q32	NN	O	O
-	NN	O	O
q41	NN	O	O
and	NN	O	O
possibly	NN	O	O
identify	NN	O	O
unlinked	NN	O	O
pedigrees	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
to	NN	O	O
6	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
(	NN	O	O
D1S249	NN	O	O
,	NN	O	O
D1S425	NN	O	O
,	NN	O	O
D1S491	NN	O	O
,	NN	O	O
D1S205	NN	O	O
,	NN	O	O
D1S414	NN	O	O
,	NN	O	O
D1S425	NN	O	O
)	NN	O	O
,	NN	O	O
yielding	NN	O	O
a	NN	O	O
maximum	NN	O	O
cumulative	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
Z	NN	O	O
=	NN	O	O
3	NN	O	O
.	NN	O	O

27	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

00	NN	O	O
for	NN	O	O
D1S245	NN	O	O
.	NN	O	O

The	NN	O	O
innermost	NN	O	O
four	NN	O	O
markers	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
one	NN	O	O
another	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
recombination	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
support	NN	O	O
linkage	NN	O	O
of	NN	O	O
vWS	NN	O	B-Disease
within	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
tightly	NN	O	O
linked	NN	O	O
markers	NN	O	O
and	NN	O	O
do	NN	O	O
not	NN	O	O
favour	NN	O	O
locus	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
trait	NN	O	O
.	NN	O	O

Null	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
ATP7B	NN	O	O
(	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
)	NN	O	O
gene	NN	O	O
results	NN	O	O
in	NN	O	O
intracellular	NN	O	B-Disease
copper	NN	O	I-Disease
accumulation	NN	O	I-Disease
and	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
onset	NN	O	I-Disease
hepatic	NN	O	I-Disease
nodular	NN	O	I-Disease
transformation	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Atp7b	NN	O	O
protein	NN	O	O
is	NN	O	O
a	NN	O	O
copper	NN	O	O
-	NN	O	O
transporting	NN	O	O
ATPase	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
in	NN	O	O
most	NN	O	O
other	NN	O	O
tissues	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATP7B	NN	O	O
gene	NN	O	O
lead	NN	O	O
to	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
copper	NN	O	B-Disease
toxicity	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
dramatic	NN	O	O
build	NN	O	O
-	NN	O	O
up	NN	O	O
of	NN	O	O
intracellular	NN	O	O
hepatic	NN	O	O
copper	NN	O	O
with	NN	O	O
subsequent	NN	O	O
hepatic	NN	O	B-Disease
and	NN	O	I-Disease
neuro	NN	O	I-Disease
-	NN	O	I-Disease
logical	NN	O	I-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

Using	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
to	NN	O	O
disrupt	NN	O	O
the	NN	O	O
normal	NN	O	O
translation	NN	O	O
of	NN	O	O
ATP7B	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
strain	NN	O	O
of	NN	O	O
mice	NN	O	O
that	NN	O	O
are	NN	O	O
homozygous	NN	O	O
mutants	NN	O	O
(	NN	O	O
null	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
ATP7B	NN	O	O
null	NN	O	O
mice	NN	O	O
display	NN	O	O
a	NN	O	O
gradual	NN	O	O
accumulation	NN	O	O
of	NN	O	O
hepatic	NN	O	O
copper	NN	O	O
that	NN	O	O
increases	NN	O	O
to	NN	O	O
a	NN	O	O
level	NN	O	O
60	NN	O	O
-	NN	O	O
fold	NN	O	O
greater	NN	O	O
than	NN	O	O
normal	NN	O	O
by	NN	O	O
5	NN	O	O
months	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

An	NN	O	O
increase	NN	O	O
in	NN	O	O
copper	NN	O	O
concentration	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
kidney	NN	O	O
,	NN	O	O
brain	NN	O	O
,	NN	O	O
placenta	NN	O	O
and	NN	O	O
lactating	NN	O	O
mammary	NN	O	O
glands	NN	O	O
of	NN	O	O
homo	NN	O	O
-	NN	O	O
zygous	NN	O	O
mutants	NN	O	O
,	NN	O	O
although	NN	O	O
milk	NN	O	O
from	NN	O	O
the	NN	O	O
mutant	NN	O	O
glands	NN	O	O
was	NN	O	O
copper	NN	O	B-Disease
deficient	NN	O	I-Disease
.	NN	O	O

Morphological	NN	O	B-Disease
abnormalities	NN	O	I-Disease
resembling	NN	O	O
cirrhosis	NN	O	B-Disease
developed	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
livers	NN	O	O
from	NN	O	O
homozygous	NN	O	O
mutants	NN	O	O
older	NN	O	O
than	NN	O	O
7	NN	O	O
months	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

Progeny	NN	O	O
of	NN	O	O
the	NN	O	O
homozygous	NN	O	O
mutant	NN	O	O
females	NN	O	O
demonstrated	NN	O	O
neurological	NN	O	B-Disease
abnormalities	NN	O	I-Disease
and	NN	O	O
growth	NN	O	B-Disease
retardation	NN	O	I-Disease
characteristic	NN	O	O
of	NN	O	O
copper	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Copper	NN	O	O
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
livers	NN	O	O
of	NN	O	O
the	NN	O	O
newborn	NN	O	O
homozygous	NN	O	O
null	NN	O	O
mutants	NN	O	O
was	NN	O	O
decreased	NN	O	O
dramatically	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
ATP7B	NN	O	O
gene	NN	O	O
produces	NN	O	O
a	NN	O	O
form	NN	O	O
of	NN	O	O
cirrhotic	NN	O	B-Disease
liver	NN	O	I-Disease
disease	NN	O	I-Disease
that	NN	O	O
resembles	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
humans	NN	O	O
and	NN	O	O
the	NN	O	O
toxic	NN	O	O
milk	NN	O	O
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
.	NN	O	O
.	NN	O	O

French	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
patients	NN	O	O
do	NN	O	O
not	NN	O	O
exhibit	NN	O	O
gametic	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
:	NN	O	O
a	NN	O	O
sperm	NN	O	O
typing	NN	O	O
analysis	NN	O	O
.	NN	O	O

Segregation	NN	O	O
distortion	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
trinucleotide	NN	O	B-Disease
repeat	NN	O	I-Disease
disorders	NN	O	I-Disease
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
a	NN	O	O
sperm	NN	O	O
typing	NN	O	O
study	NN	O	O
performed	NN	O	O
in	NN	O	O
patients	NN	O	O
of	NN	O	O
Japanese	NN	O	O
descent	NN	O	O
with	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
reported	NN	O	O
that	NN	O	O
disease	NN	O	O
alleles	NN	O	O
are	NN	O	O
preferentially	NN	O	O
transmitted	NN	O	O
during	NN	O	O
meiosis	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
a	NN	O	O
sperm	NN	O	O
typing	NN	O	O
study	NN	O	O
of	NN	O	O
five	NN	O	O
MJD	NN	O	B-Disease
patients	NN	O	O
of	NN	O	O
French	NN	O	O
descent	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
pooled	NN	O	O
data	NN	O	O
shows	NN	O	O
a	NN	O	O
ratio	NN	O	O
of	NN	O	O
mutant	NN	O	O
to	NN	O	O
normal	NN	O	O
alleles	NN	O	O
of	NN	O	O
379	NN	O	O
436	NN	O	O
(	NN	O	O
46	NN	O	O
.	NN	O	O
5	NN	O	O
53	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
support	NN	O	O
meiotic	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
.	NN	O	O

To	NN	O	O
confirm	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
sperm	NN	O	O
typing	NN	O	O
analysis	NN	O	O
was	NN	O	O
also	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
polymorphic	NN	O	O
marker	NN	O	O
,	NN	O	O
D14S1050	NN	O	O
,	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
.	NN	O	O

Among	NN	O	O
910	NN	O	O
sperm	NN	O	O
analyzed	NN	O	O
,	NN	O	O
the	NN	O	O
allele	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
disease	NN	O	O
chromosome	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
50	NN	O	O
.	NN	O	O

3	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
samples	NN	O	O
and	NN	O	O
the	NN	O	O
allele	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
chromosome	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
49	NN	O	O
.	NN	O	O

6	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
sperm	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
frequency	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
alleles	NN	O	O
is	NN	O	O
not	NN	O	O
significant	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
8423	NN	O	O
)	NN	O	O
.	NN	O	O

Likelihood	NN	O	O
-	NN	O	O
based	NN	O	O
analysis	NN	O	O
of	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
in	NN	O	O
the	NN	O	O
single	NN	O	O
sperm	NN	O	O
data	NN	O	O
using	NN	O	O
the	NN	O	O
SPERMSEG	NN	O	O
program	NN	O	O
also	NN	O	O
showed	NN	O	O
no	NN	O	O
support	NN	O	O
for	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
at	NN	O	O
the	NN	O	O
gamete	NN	O	O
level	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
previous	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
also	NN	O	O
suggested	NN	O	O
that	NN	O	O
disease	NN	O	O
allele	NN	O	O
stability	NN	O	O
may	NN	O	O
be	NN	O	O
influenced	NN	O	O
by	NN	O	O
a	NN	O	O
trans	NN	O	O
effect	NN	O	O
of	NN	O	O
an	NN	O	O
intragenic	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
987	NN	O	O
G	NN	O	O
/	NN	O	O
C	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
allele	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
French	NN	O	O
patients	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
this	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
variance	NN	O	O
in	NN	O	O
repeat	NN	O	O
number	NN	O	O
in	NN	O	O
sperm	NN	O	O
from	NN	O	O
the	NN	O	O
French	NN	O	O
MJD	NN	O	B-Disease
patients	NN	O	O
overlapped	NN	O	O
significantly	NN	O	O
with	NN	O	O
the	NN	O	O
variance	NN	O	O
in	NN	O	O
repeat	NN	O	O
number	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
C	NN	O	O
/	NN	O	O
C	NN	O	O
homozygous	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O

Missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
alternative	NN	O	O
splice	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
in	NN	O	O
eye	NN	O	B-Disease
anomalies	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
ocular	NN	O	O
morphogenesis	NN	O	O
,	NN	O	O
and	NN	O	O
PAX6	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
ocular	NN	O	B-Disease
anomalies	NN	O	I-Disease
,	NN	O	O
including	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
,	NN	O	O
corneal	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
congenital	NN	O	B-Disease
cataract	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
foveal	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
that	NN	O	O
recognizes	NN	O	O
target	NN	O	O
genes	NN	O	O
through	NN	O	O
its	NN	O	O
paired	NN	O	O
-	NN	O	O
type	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
paired	NN	O	O
domain	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
two	NN	O	O
distinct	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
subdomains	NN	O	O
,	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
subdomain	NN	O	O
(	NN	O	O
NTS	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
subdomain	NN	O	O
(	NN	O	O
CTS	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
bind	NN	O	O
respective	NN	O	O
consensus	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
produces	NN	O	O
two	NN	O	O
alternative	NN	O	O
splice	NN	O	O
isoforms	NN	O	O
that	NN	O	O
have	NN	O	O
the	NN	O	O
distinct	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
paired	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
insertion	NN	O	O
,	NN	O	O
into	NN	O	O
the	NN	O	O
NTS	NN	O	O
,	NN	O	O
of	NN	O	O
14	NN	O	O
additional	NN	O	O
amino	NN	O	O
acids	NN	O	O
encoded	NN	O	O
by	NN	O	O
exon	NN	O	O
5a	NN	O	O
abolishes	NN	O	O
the	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NTS	NN	O	O
and	NN	O	O
unmasks	NN	O	O
the	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
CTS	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
exon	NN	O	O
5a	NN	O	O
appears	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
molecular	NN	O	O
switch	NN	O	O
that	NN	O	O
specifies	NN	O	O
target	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
ascertained	NN	O	O
a	NN	O	O
novel	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
four	NN	O	O
pedigrees	NN	O	O
with	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
,	NN	O	O
congenital	NN	O	B-Disease
cataract	NN	O	I-Disease
,	NN	O	O
Axenfeldt	NN	O	B-Disease
anomaly	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
foveal	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
,	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
mutation	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
splice	NN	O	O
-	NN	O	O
variant	NN	O	O
region	NN	O	O
.	NN	O	O

A	NN	O	O
T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
20th	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
of	NN	O	O
exon	NN	O	O
5a	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Asp	NN	O	O
(	NN	O	O
GTC	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
GAC	NN	O	O
)	NN	O	O
substitution	NN	O	O
at	NN	O	O
the	NN	O	O
7th	NN	O	O
codon	NN	O	O
of	NN	O	O
the	NN	O	O
alternative	NN	O	O
splice	NN	O	O
region	NN	O	O
.	NN	O	O

Functional	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
V54D	NN	O	O
mutation	NN	O	O
slightly	NN	O	O
increased	NN	O	O
NTS	NN	O	O
binding	NN	O	O
and	NN	O	O
decreased	NN	O	O
CTS	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
to	NN	O	O
almost	NN	O	O
half	NN	O	O
.	NN	O	O
.	NN	O	O

Penetrances	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
1675delA	NN	O	O
and	NN	O	O
1135insA	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
and	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

For	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
and	NN	O	O
predictive	NN	O	O
testing	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
inherited	NN	O	B-Disease
breast	NN	O	I-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
penetrances	NN	O	O
and	NN	O	O
expressions	NN	O	O
of	NN	O	O
the	NN	O	O
underlying	NN	O	O
mutations	NN	O	O
should	NN	O	O
be	NN	O	O
known	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
two	NN	O	O
BRCA1	NN	O	O
founder	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Norwegian	NN	O	O
population	NN	O	O
.	NN	O	O

Index	NN	O	O
cases	NN	O	O
for	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
were	NN	O	O
found	NN	O	O
two	NN	O	O
different	NN	O	O
ways	NN	O	O
through	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
consecutive	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
(	NN	O	O
n	NN	O	O
=	NN	O	O
16	NN	O	O
)	NN	O	O
and	NN	O	O
through	NN	O	O
our	NN	O	O
family	NN	O	O
cancer	NN	O	B-Disease
clinic	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
14	NN	O	O
)	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
20	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
had	NN	O	O
BRCA1	NN	O	O
1675delA	NN	O	O
,	NN	O	O
and	NN	O	O
10	NN	O	O
had	NN	O	O
1135insA	NN	O	O
.	NN	O	O

Their	NN	O	O
relatives	NN	O	O
were	NN	O	O
described	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
absence	NN	O	O
/	NN	O	O
presence	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Of	NN	O	O
133	NN	O	O
living	NN	O	O
female	NN	O	O
relatives	NN	O	O
,	NN	O	O
83	NN	O	O
(	NN	O	O
62	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
,	NN	O	O
in	NN	O	O
penetrance	NN	O	O
and	NN	O	O
expression	NN	O	O
,	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
whereas	NN	O	O
differences	NN	O	O
according	NN	O	O
to	NN	O	O
method	NN	O	O
of	NN	O	O
ascertainment	NN	O	O
were	NN	O	O
seen	NN	O	O
.	NN	O	O

The	NN	O	O
overall	NN	O	O
findings	NN	O	O
were	NN	O	O
that	NN	O	O
disease	NN	O	O
started	NN	O	O
to	NN	O	O
occur	NN	O	O
at	NN	O	O
age	NN	O	O
30	NN	O	O
years	NN	O	O
and	NN	O	O
that	NN	O	O
by	NN	O	O
age	NN	O	O
50	NN	O	O
years	NN	O	O
48	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
-	NN	O	O
carrying	NN	O	O
women	NN	O	O
had	NN	O	O
experienced	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

More	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
than	NN	O	O
breast	NN	O	B-Disease
cancers	NN	O	I-Disease
were	NN	O	O
recorded	NN	O	O
.	NN	O	O

Both	NN	O	O
penetrance	NN	O	O
and	NN	O	O
expression	NN	O	O
(	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
vs	NN	O	O
.	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
)	NN	O	O
were	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
reports	NN	O	O
of	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
founder	NN	O	O
mutations	NN	O	O
.	NN	O	O

Whether	NN	O	O
the	NN	O	O
reported	NN	O	O
differences	NN	O	O
reflect	NN	O	O
true	NN	O	O
differences	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
methodological	NN	O	O
problems	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

An	NN	O	O
observed	NN	O	O
excess	NN	O	O
of	NN	O	O
mutation	NN	O	O
carriers	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
methodological	NN	O	O
problems	NN	O	O
;	NN	O	O
possible	NN	O	O
explanations	NN	O	O
were	NN	O	O
a	NN	O	O
"	NN	O	O
true	NN	O	O
"	NN	O	O
low	NN	O	O
penetrance	NN	O	O
or	NN	O	O
preferential	NN	O	O
segregation	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
dermatofibrosarcoma	NN	O	B-Disease
protuberans	NN	O	I-Disease
-	NN	O	O
associated	NN	O	O
collagen	NN	O	O
type	NN	O	O
Ialpha1	NN	O	O
/	NN	O	O
platelet	NN	O	O
-	NN	O	O
derived	NN	O	O
growth	NN	O	O
factor	NN	O	O
(	NN	O	O
PDGF	NN	O	O
)	NN	O	O
B	NN	O	O
-	NN	O	O
chain	NN	O	O
fusion	NN	O	O
gene	NN	O	O
generates	NN	O	O
a	NN	O	O
transforming	NN	O	O
protein	NN	O	O
that	NN	O	O
is	NN	O	O
processed	NN	O	O
to	NN	O	O
functional	NN	O	O
PDGF	NN	O	O
-	NN	O	O
BB	NN	O	O
.	NN	O	O

Dermatofibrosarcoma	NN	O	B-Disease
protuberans	NN	O	I-Disease
(	NN	O	O
DFSP	NN	O	B-Disease
)	NN	O	O
displays	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
involving	NN	O	O
chromosome	NN	O	O
17	NN	O	O
and	NN	O	O
22	NN	O	O
,	NN	O	O
which	NN	O	O
fuse	NN	O	O
the	NN	O	O
collagen	NN	O	O
type	NN	O	O
Ialpha1	NN	O	O
(	NN	O	O
COLIA1	NN	O	O
)	NN	O	O
gene	NN	O	O
to	NN	O	O
the	NN	O	O
platelet	NN	O	O
-	NN	O	O
derived	NN	O	O
growth	NN	O	O
factor	NN	O	O
(	NN	O	O
PDGF	NN	O	O
)	NN	O	O
B	NN	O	O
-	NN	O	O
chain	NN	O	O
(	NN	O	O
PDGFB	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
functional	NN	O	O
and	NN	O	O
structural	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
fusion	NN	O	O
protein	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
stable	NN	O	O
NIH3T3	NN	O	O
cell	NN	O	O
line	NN	O	O
that	NN	O	O
contained	NN	O	O
a	NN	O	O
tumor	NN	O	O
-	NN	O	O
derived	NN	O	O
chimeric	NN	O	O
gene	NN	O	O
resulting	NN	O	O
from	NN	O	O
a	NN	O	O
COIA1	NN	O	O
intron	NN	O	O
7	NN	O	O
-	NN	O	O
PDGFB	NN	O	O
intron	NN	O	O
1	NN	O	O
fusion	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
fusion	NN	O	O
protein	NN	O	O
led	NN	O	O
to	NN	O	O
morphological	NN	O	O
transformation	NN	O	O
and	NN	O	O
increased	NN	O	O
growth	NN	O	O
rate	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
PDGF	NN	O	O
receptor	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
CGP57148B	NN	O	O
reversed	NN	O	O
the	NN	O	O
transformed	NN	O	O
phenotype	NN	O	O
and	NN	O	O
reduced	NN	O	O
the	NN	O	O
growth	NN	O	O
rate	NN	O	O
of	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
-	NN	O	O
expressing	NN	O	O
cells	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
effects	NN	O	O
on	NN	O	O
control	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
dimeric	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
precursors	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
through	NN	O	O
PDGFB	NN	O	O
immunoprecipitations	NN	O	O
of	NN	O	O
metabolically	NN	O	O
labeled	NN	O	O
cells	NN	O	O
and	NN	O	O
also	NN	O	O
by	NN	O	O
PDGFB	NN	O	O
immunoprecipitations	NN	O	O
followed	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
with	NN	O	O
COLIA1	NN	O	O
antibodies	NN	O	O
.	NN	O	O

Pulse	NN	O	O
-	NN	O	O
chase	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
precursor	NN	O	O
was	NN	O	O
processed	NN	O	O
to	NN	O	O
an	NN	O	O
end	NN	O	O
product	NN	O	O
that	NN	O	O
was	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
PDGF	NN	O	O
-	NN	O	O
BB	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
-	NN	O	O
expressing	NN	O	O
cells	NN	O	O
generated	NN	O	O
tumors	NN	O	B-Disease
after	NN	O	O
s	NN	O	O
.	NN	O	O

c	NN	O	O
c	NN	O	O
.	NN	O	O
injection	NN	O	O
into	NN	O	O
nude	NN	O	O
mice	NN	O	O
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-Disease
growth	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
CGP57148B	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
COLIA1	NN	O	O
/	NN	O	O
PDGFB	NN	O	O
fusion	NN	O	O
associated	NN	O	O
with	NN	O	O
DFSP	NN	O	B-Disease
contributes	NN	O	O
to	NN	O	O
tumor	NN	O	B-Disease
development	NN	O	O
through	NN	O	O
ectopic	NN	O	O
production	NN	O	O
of	NN	O	O
PDGF	NN	O	O
-	NN	O	O
BB	NN	O	O
and	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
an	NN	O	O
autocrine	NN	O	O
loop	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
,	NN	O	O
thus	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
PDGF	NN	O	O
receptors	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
pharmacological	NN	O	O
treatment	NN	O	O
of	NN	O	O
DFSP	NN	O	B-Disease
and	NN	O	O
giant	NN	O	B-Disease
cell	NN	O	I-Disease
fibroblastoma	NN	O	I-Disease
,	NN	O	O
e	NN	O	O
.	NN	O	O
g	NN	O	O
.	NN	O	O
,	NN	O	O
through	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
PDGF	NN	O	O
receptor	NN	O	O
kinase	NN	O	O
inhibitors	NN	O	O
such	NN	O	O
as	NN	O	O
CGP57148B	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
PEX1	NN	O	O
mutation	NN	O	O
in	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Zellweger	NN	O	B-Disease
spectrum	NN	O	I-Disease
of	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
encompassing	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
progressively	NN	O	O
milder	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
neonatal	NN	O	B-Disease
adrenoleukodystrophy	NN	O	I-Disease
and	NN	O	O
infantile	NN	O	B-Disease
Refsum	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
form	NN	O	O
functional	NN	O	O
peroxisomes	NN	O	O
.	NN	O	O

Cell	NN	O	O
fusion	NN	O	O
complementation	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
these	NN	O	O
diseases	NN	O	O
are	NN	O	O
genetically	NN	O	O
heterogeneous	NN	O	O
,	NN	O	O
with	NN	O	O
two	NN	O	O
-	NN	O	O
thirds	NN	O	O
of	NN	O	O
all	NN	O	O
patients	NN	O	O
lying	NN	O	O
within	NN	O	O
a	NN	O	O
single	NN	O	O
complementation	NN	O	O
group	NN	O	O
,	NN	O	O
CG1	NN	O	O
.	NN	O	O

Molecular	NN	O	O
genetic	NN	O	O
and	NN	O	O
cell	NN	O	O
biology	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
PEX1	NN	O	O
is	NN	O	O
deficient	NN	O	O
in	NN	O	O
many	NN	O	O
CG1	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
previous	NN	O	O
studies	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
mildly	NN	O	O
affected	NN	O	O
patients	NN	O	O
and	NN	O	O
there	NN	O	O
is	NN	O	O
still	NN	O	O
no	NN	O	O
report	NN	O	O
of	NN	O	O
two	NN	O	O
mutant	NN	O	O
PEX1	NN	O	O
alleles	NN	O	O
in	NN	O	O
any	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
patient	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PMP70	NN	O	O
gene	NN	O	O
have	NN	O	O
also	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
two	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
patients	NN	O	O
from	NN	O	O
CG1	NN	O	O
,	NN	O	O
raising	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
CG1	NN	O	O
patients	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
PEX1	NN	O	O
-	NN	O	O
deficient	NN	O	O
and	NN	O	O
PMP70	NN	O	O
-	NN	O	O
deficient	NN	O	O
individuals	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
disease	NN	O	O
in	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
patients	NN	O	O
from	NN	O	O
CG1	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
all	NN	O	O
24	NN	O	O
PEX1	NN	O	O
exons	NN	O	O
in	NN	O	O
four	NN	O	O
patients	NN	O	O
,	NN	O	O
including	NN	O	O
both	NN	O	O
patients	NN	O	O
that	NN	O	O
have	NN	O	O
mutations	NN	O	O
in	NN	O	O
PMP70	NN	O	O
.	NN	O	O

PEX1	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
four	NN	O	O
patients	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
1	NN	O	O
-	NN	O	O
bp	NN	O	O
insertion	NN	O	O
(	NN	O	O
c	NN	O	O
.	NN	O	O
2097insT	NN	O	O
)	NN	O	O
in	NN	O	O
exon	NN	O	O
13	NN	O	O
that	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
three	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
patients	NN	O	O
.	NN	O	O

Subsequent	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
one	NN	O	O
-	NN	O	O
half	NN	O	O
of	NN	O	O
all	NN	O	O
CG1	NN	O	O
patients	NN	O	O
and	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
phenotype	NN	O	O
.	NN	O	O

As	NN	O	O
this	NN	O	O
mutation	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
loss	NN	O	O
of	NN	O	O
protein	NN	O	O
function	NN	O	O
its	NN	O	O
frequency	NN	O	O
makes	NN	O	O
it	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
cause	NN	O	O
of	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
helping	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
high	NN	O	O
percentage	NN	O	O
of	NN	O	O
patients	NN	O	O
that	NN	O	O
belong	NN	O	O
to	NN	O	O
CG1	NN	O	O
.	NN	O	O

Novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
protein	NN	O	O
gene	NN	O	O
and	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
,	NN	O	O
translational	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
immunodeficiency	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
thrombocytopenia	NN	O	B-Disease
,	NN	O	O
eczema	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
recurrent	NN	O	O
infections	NN	O	O
,	NN	O	O
and	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
protein	NN	O	O
(	NN	O	O
WASP	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

WASP	NN	O	O
contains	NN	O	O
several	NN	O	O
functional	NN	O	O
domains	NN	O	O
through	NN	O	O
which	NN	O	O
it	NN	O	O
interacts	NN	O	O
with	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
actin	NN	O	O
cytoskeleton	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
17	NN	O	O
WASP	NN	O	O
gene	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
12	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
novel	NN	O	O
.	NN	O	O

DNA	NN	O	O
of	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
was	NN	O	O
PCR	NN	O	O
amplified	NN	O	O
and	NN	O	O
analyzed	NN	O	O
by	NN	O	O
SSCA	NN	O	O
,	NN	O	O
heteroduplex	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
level	NN	O	O
were	NN	O	O
ascertained	NN	O	O
by	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analyses	NN	O	O
.	NN	O	O

All	NN	O	O
missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
located	NN	O	O
in	NN	O	O
exons	NN	O	O
1	NN	O	O
-	NN	O	O
4	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
the	NN	O	O
nonsense	NN	O	O
,	NN	O	O
frameshift	NN	O	O
and	NN	O	O
splice	NN	O	O
site	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
exons	NN	O	O
6	NN	O	O
-	NN	O	O
11	NN	O	O
.	NN	O	O

Mutations	NN	O	O
that	NN	O	O
alter	NN	O	O
splice	NN	O	O
sites	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
several	NN	O	O
types	NN	O	O
of	NN	O	O
mRNAs	NN	O	O
,	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
which	NN	O	O
represented	NN	O	O
the	NN	O	O
normally	NN	O	O
spliced	NN	O	O
product	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
normally	NN	O	O
spliced	NN	O	O
transcripts	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
milder	NN	O	O
phenotype	NN	O	O
.	NN	O	O

When	NN	O	O
one	NN	O	O
such	NN	O	O
case	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
,	NN	O	O
reduced	NN	O	O
amounts	NN	O	O
of	NN	O	O
normal	NN	O	O
-	NN	O	O
size	NN	O	O
WASP	NN	O	O
were	NN	O	O
present	NN	O	O
.	NN	O	O

In	NN	O	O
other	NN	O	O
cases	NN	O	O
as	NN	O	O
well	NN	O	O
,	NN	O	O
a	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
normal	NN	O	O
or	NN	O	O
mutant	NN	O	O
WASP	NN	O	O
present	NN	O	O
and	NN	O	O
the	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
individuals	NN	O	O
.	NN	O	O

No	NN	O	O
protein	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
two	NN	O	O
individuals	NN	O	O
with	NN	O	O
severe	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O

Reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
a	NN	O	O
normal	NN	O	O
-	NN	O	O
size	NN	O	O
WASP	NN	O	O
with	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
two	NN	O	O
individuals	NN	O	O
with	NN	O	O
XLT	NN	O	B-Disease
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
at	NN	O	O
the	NN	O	O
DNA	NN	O	O
level	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
predicting	NN	O	O
clinical	NN	O	O
course	NN	O	O
.	NN	O	O

Studies	NN	O	O
at	NN	O	O
the	NN	O	O
transcript	NN	O	O
and	NN	O	O
protein	NN	O	O
level	NN	O	O
are	NN	O	O
needed	NN	O	O
for	NN	O	O
a	NN	O	O
better	NN	O	O
assessment	NN	O	O
.	NN	O	O
.	NN	O	O

Aminoglycoside	NN	O	O
antibiotics	NN	O	O
restore	NN	O	O
dystrophin	NN	O	O
function	NN	O	O
to	NN	O	O
skeletal	NN	O	O
muscles	NN	O	O
of	NN	O	O
mdx	NN	O	O
mice	NN	O	O
.	NN	O	O

Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
protein	NN	O	O
in	NN	O	O
striated	NN	O	O
muscle	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
number	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
are	NN	O	O
premature	NN	O	O
stop	NN	O	O
codons	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
aminoglycoside	NN	O	O
treatment	NN	O	O
can	NN	O	O
suppress	NN	O	O
stop	NN	O	O
codons	NN	O	O
in	NN	O	O
cultured	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
gentamicin	NN	O	O
on	NN	O	O
cultured	NN	O	O
muscle	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
mdx	NN	O	O
mouse	NN	O	O
-	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
that	NN	O	O
possesses	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
in	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
mdx	NN	O	O
myotubes	NN	O	O
to	NN	O	O
gentamicin	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
localization	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
evaluated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
differing	NN	O	O
dosages	NN	O	O
of	NN	O	O
gentamicin	NN	O	O
on	NN	O	O
expression	NN	O	O
and	NN	O	O
functional	NN	O	O
protection	NN	O	O
of	NN	O	O
the	NN	O	O
muscles	NN	O	O
of	NN	O	O
mdx	NN	O	O
mice	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
treatment	NN	O	O
regimen	NN	O	O
that	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
in	NN	O	O
all	NN	O	O
striated	NN	O	O
muscles	NN	O	O
examined	NN	O	O
and	NN	O	O
that	NN	O	O
provided	NN	O	O
functional	NN	O	O
protection	NN	O	O
against	NN	O	O
muscular	NN	O	B-Disease
injury	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
aminoglycosides	NN	O	O
can	NN	O	O
suppress	NN	O	O
stop	NN	O	O
codons	NN	O	O
not	NN	O	O
only	NN	O	O
in	NN	O	O
vitro	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
raise	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
treatment	NN	O	O
regimen	NN	O	O
for	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
other	NN	O	O
diseases	NN	O	O
caused	NN	O	O
by	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
mutations	NN	O	O
.	NN	O	O

This	NN	O	O
treatment	NN	O	O
could	NN	O	O
prove	NN	O	O
effective	NN	O	O
in	NN	O	O
up	NN	O	O
to	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
the	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
gene	NN	O	O
product	NN	O	O
causes	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
in	NN	O	O
target	NN	O	O
organs	NN	O	O
.	NN	O	O

Ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
markedly	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
,	NN	O	O
increased	NN	O	O
incidence	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
neurodegeneration	NN	O	B-Disease
,	NN	O	O
especially	NN	O	O
of	NN	O	O
the	NN	O	O
cerebellar	NN	O	O
Purkinje	NN	O	O
cells	NN	O	O
.	NN	O	O

Ionizing	NN	O	O
radiation	NN	O	O
oxidizes	NN	O	O
macromolecules	NN	O	O
and	NN	O	O
causes	NN	O	O
tissue	NN	O	O
damage	NN	O	O
through	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
is	NN	O	O
due	NN	O	O
to	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
resulting	NN	O	O
from	NN	O	O
loss	NN	O	O
of	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
gene	NN	O	O
product	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
employed	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
of	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
mouse	NN	O	O
with	NN	O	O
a	NN	O	O
disrupted	NN	O	O
Atm	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
organs	NN	O	O
which	NN	O	O
develop	NN	O	O
pathologic	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
Atm	NN	O	O
-	NN	O	O
deficient	NN	O	O
mice	NN	O	O
are	NN	O	O
targets	NN	O	O
of	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
cerebellar	NN	O	O
Purkinje	NN	O	O
cells	NN	O	O
are	NN	O	O
particularly	NN	O	O
affected	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
provide	NN	O	O
a	NN	O	O
mechanistic	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
phenotype	NN	O	O
and	NN	O	O
lay	NN	O	O
a	NN	O	O
rational	NN	O	O
foundation	NN	O	O
for	NN	O	O
therapeutic	NN	O	O
intervention	NN	O	O
.	NN	O	O
.	NN	O	O

Recessively	NN	O	O
inherited	NN	O	O
multiple	NN	O	B-Disease
epiphyseal	NN	O	I-Disease
dysplasia	NN	O	I-Disease
with	NN	O	O
normal	NN	O	O
stature	NN	O	O
,	NN	O	O
club	NN	O	B-Disease
foot	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
double	NN	O	B-Disease
layered	NN	O	I-Disease
patella	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
DTDST	NN	O	O
mutation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
observed	NN	O	O
over	NN	O	O
25	NN	O	O
different	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
sulphate	NN	O	O
transporter	NN	O	O
gene	NN	O	O
(	NN	O	O
DTDST	NN	O	O
)	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
the	NN	O	O
recessive	NN	O	B-Disease
disorders	NN	O	I-Disease
achondrogenesis	NN	O	B-Disease
1B	NN	O	I-Disease
,	NN	O	O
atelosteogenesis	NN	O	B-Disease
2	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
c862t	NN	O	O
(	NN	O	O
R279W	NN	O	O
)	NN	O	O
transition	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Finnish	NN	O	O
patients	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
these	NN	O	O
disorders	NN	O	O
it	NN	O	O
is	NN	O	O
usually	NN	O	O
combined	NN	O	O
with	NN	O	O
other	NN	O	O
DTDST	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
had	NN	O	O
not	NN	O	O
seen	NN	O	O
a	NN	O	O
case	NN	O	O
of	NN	O	O
homozygosity	NN	O	O
for	NN	O	O
c862t	NN	O	O
(	NN	O	O
R279W	NN	O	O
)	NN	O	O
until	NN	O	O
we	NN	O	O
analysed	NN	O	O
DNA	NN	O	O
from	NN	O	O
a	NN	O	O
36	NN	O	O
year	NN	O	O
old	NN	O	O
male	NN	O	O
with	NN	O	O
tall	NN	O	O
-	NN	O	O
normal	NN	O	O
stature	NN	O	O
(	NN	O	O
180	NN	O	O
cm	NN	O	O
)	NN	O	O
who	NN	O	O
asked	NN	O	O
for	NN	O	O
genetic	NN	O	O
counselling	NN	O	O
for	NN	O	O
suspected	NN	O	O
multiple	NN	O	B-Disease
epiphyseal	NN	O	I-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

He	NN	O	O
was	NN	O	O
treated	NN	O	O
for	NN	O	O
club	NN	O	B-Disease
foot	NN	O	I-Disease
and	NN	O	O
hip	NN	O	B-Disease
dysplasia	NN	O	I-Disease
at	NN	O	O
birth	NN	O	O
.	NN	O	O

Skeletal	NN	O	O
changes	NN	O	O
consistent	NN	O	O
with	NN	O	O
multiple	NN	O	B-Disease
epiphyseal	NN	O	I-Disease
dysplasia	NN	O	I-Disease
,	NN	O	O
with	NN	O	O
the	NN	O	O
peculiar	NN	O	O
finding	NN	O	O
of	NN	O	O
a	NN	O	O
double	NN	O	B-Disease
layered	NN	O	I-Disease
patella	NN	O	I-Disease
,	NN	O	O
were	NN	O	O
recognised	NN	O	O
during	NN	O	O
childhood	NN	O	O
.	NN	O	O

Cleft	NN	O	B-Disease
palate	NN	O	I-Disease
,	NN	O	O
swelling	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
ear	NN	O	I-Disease
pinna	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hitch	NN	O	B-Disease
hiker	NN	O	I-Disease
thumb	NN	O	I-Disease
were	NN	O	O
absent	NN	O	O
.	NN	O	O

He	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
homozygous	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
healthy	NN	O	O
parents	NN	O	O
heterozygous	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
R279W	NN	O	O
mutation	NN	O	O
in	NN	O	O
DTDST	NN	O	O
,	NN	O	O
and	NN	O	O
his	NN	O	O
fibroblasts	NN	O	O
showed	NN	O	O
a	NN	O	O
sulphate	NN	O	O
incorporation	NN	O	O
defect	NN	O	O
typical	NN	O	O
of	NN	O	O
DTDST	NN	O	B-Disease
disorders	NN	O	I-Disease
.	NN	O	O

Counselling	NN	O	O
was	NN	O	O
given	NN	O	O
for	NN	O	O
a	NN	O	O
recessive	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
thereby	NN	O	O
considerably	NN	O	O
reducing	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
affected	NN	O	O
offspring	NN	O	O
.	NN	O	O

Multiple	NN	O	B-Disease
epiphyseal	NN	O	I-Disease
dysplasia	NN	O	I-Disease
is	NN	O	O
more	NN	O	O
frequently	NN	O	O
caused	NN	O	O
by	NN	O	O
dominant	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
COMP	NN	O	O
(	NN	O	O
EDM1	NN	O	B-Disease
,	NN	O	O
McKusick	NN	O	O
132400	NN	O	O
)	NN	O	O
and	NN	O	O
COL9A2	NN	O	O
genes	NN	O	O
(	NN	O	O
EDM2	NN	O	B-Disease
,	NN	O	O
McKusick	NN	O	O
600204	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
few	NN	O	O
other	NN	O	O
patients	NN	O	O
and	NN	O	O
families	NN	O	O
with	NN	O	O
features	NN	O	O
similar	NN	O	O
to	NN	O	O
our	NN	O	O
proband	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
previously	NN	O	O
and	NN	O	O
considered	NN	O	O
to	NN	O	O
have	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
MED	NN	O	B-Disease
(	NN	O	O
EDM4	NN	O	B-Disease
,	NN	O	O
McKusick	NN	O	O
226900	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
confirms	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
this	NN	O	O
entity	NN	O	O
and	NN	O	O
assigns	NN	O	O
it	NN	O	O
to	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
spectrum	NN	O	O
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
DTDST	NN	O	O
locus	NN	O	O
.	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
for	NN	O	O
a	NN	O	O
novel	NN	O	O
DTDST	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
a	NN	O	O
'	NN	O	O
broad	NN	O	O
bone	NN	O	O
-	NN	O	O
platyspondylic	NN	O	O
'	NN	O	O
variant	NN	O	O
of	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

Atypical	NN	O	O
or	NN	O	O
variant	NN	O	O
forms	NN	O	O
of	NN	O	O
well	NN	O	O
-	NN	O	O
known	NN	O	O
chondrodysplasias	NN	O	B-Disease
may	NN	O	O
pose	NN	O	O
diagnostic	NN	O	O
problems	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
girl	NN	O	O
with	NN	O	O
clinical	NN	O	O
features	NN	O	O
suggesting	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
but	NN	O	O
with	NN	O	O
unusual	NN	O	O
radiographic	NN	O	O
features	NN	O	O
including	NN	O	O
severe	NN	O	O
platyspondyly	NN	O	B-Disease
,	NN	O	O
wide	NN	O	O
metaphyses	NN	O	O
,	NN	O	O
and	NN	O	O
fibular	NN	O	O
overgrowth	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
partially	NN	O	O
reminiscent	NN	O	O
of	NN	O	O
metatropic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
diagnosis	NN	O	O
was	NN	O	O
clarified	NN	O	O
by	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
DTDST	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
revealed	NN	O	O
homozygosity	NN	O	O
for	NN	O	O
a	NN	O	O
previously	NN	O	O
undescribed	NN	O	O
mutation	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
Q454P	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
10th	NN	O	O
transmembrane	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
DTDST	NN	O	O
sulfate	NN	O	O
transporter	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
particular	NN	O	O
value	NN	O	O
in	NN	O	O
such	NN	O	O
atypical	NN	O	O
cases	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
type	NN	O	O
of	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
at	NN	O	O
APC	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
is	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
germline	NN	O	O
mutation	NN	O	O
:	NN	O	O
a	NN	O	O
new	NN	O	O
facet	NN	O	O
to	NN	O	O
Knudson	NN	O	O
'	NN	O	O
s	NN	O	O
'	NN	O	O
two	NN	O	O
-	NN	O	O
hit	NN	O	O
'	NN	O	O
hypothesis	NN	O	O
.	NN	O	O

APC	NN	O	O
is	NN	O	O
often	NN	O	O
cited	NN	O	O
as	NN	O	O
a	NN	O	O
prime	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
.	NN	O	O

Truncating	NN	O	O
germline	NN	O	O
and	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
(	NN	O	O
or	NN	O	O
,	NN	O	O
infrequently	NN	O	O
,	NN	O	O
allelic	NN	O	O
loss	NN	O	O
)	NN	O	O
occur	NN	O	O
in	NN	O	O
tumors	NN	O	B-Disease
in	NN	O	O
FAP	NN	O	B-Disease
(	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

Most	NN	O	O
sporadic	NN	O	B-Disease
colorectal	NN	O	I-Disease
cancers	NN	O	I-Disease
also	NN	O	O
have	NN	O	O
two	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
.	NN	O	O

Clues	NN	O	O
from	NN	O	O
attenuated	NN	O	B-Disease
polyposis	NN	O	I-Disease
,	NN	O	O
missense	NN	O	O
germline	NN	O	O
variants	NN	O	O
with	NN	O	O
mild	NN	O	O
disease	NN	O	O
and	NN	O	O
the	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
cluster	NN	O	O
region	NN	O	O
(	NN	O	O
codons	NN	O	O
1	NN	O	O
,	NN	O	O
250	NN	O	O
-	NN	O	O
1	NN	O	O
,	NN	O	O
450	NN	O	O
)	NN	O	O
indicate	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
might	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
simple	NN	O	O
loss	NN	O	O
of	NN	O	O
protein	NN	O	O
function	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
germline	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
within	NN	O	O
a	NN	O	O
small	NN	O	O
region	NN	O	O
(	NN	O	O
codons	NN	O	O
1	NN	O	O
,	NN	O	O
194	NN	O	O
-	NN	O	O
1	NN	O	O
,	NN	O	O
392	NN	O	O
at	NN	O	O
most	NN	O	O
)	NN	O	O
mainly	NN	O	O
show	NN	O	O
allelic	NN	O	O
loss	NN	O	O
in	NN	O	O
their	NN	O	O
colorectal	NN	O	B-Disease
adenomas	NN	O	I-Disease
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
other	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
whose	NN	O	O
second	NN	O	O
hits	NN	O	O
tend	NN	O	O
to	NN	O	O
occur	NN	O	O
by	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
mutation	NN	O	O
cluster	NN	O	O
region	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
different	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
provide	NN	O	O
cells	NN	O	O
with	NN	O	O
different	NN	O	O
selective	NN	O	O
advantages	NN	O	O
,	NN	O	O
with	NN	O	O
mutations	NN	O	O
close	NN	O	O
to	NN	O	O
codon	NN	O	O
1	NN	O	O
,	NN	O	O
300	NN	O	O
providing	NN	O	O
the	NN	O	O
greatest	NN	O	O
advantage	NN	O	O
.	NN	O	O

Allelic	NN	O	O
loss	NN	O	O
is	NN	O	O
selected	NN	O	O
strongly	NN	O	O
in	NN	O	O
cells	NN	O	O
with	NN	O	O
one	NN	O	O
mutation	NN	O	O
near	NN	O	O
codon	NN	O	O
1	NN	O	O
,	NN	O	O
300	NN	O	O
.	NN	O	O

A	NN	O	O
different	NN	O	O
germline	NN	O	O
-	NN	O	O
somatic	NN	O	O
APC	NN	O	B-Disease
mutation	NN	O	O
association	NN	O	O
exists	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
desmoids	NN	O	I-Disease
.	NN	O	O

APC	NN	O	O
is	NN	O	O
not	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
a	NN	O	O
classical	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
also	NN	O	O
indicate	NN	O	O
a	NN	O	O
new	NN	O	O
mechanism	NN	O	O
for	NN	O	O
disease	NN	O	O
severity	NN	O	O
if	NN	O	O
a	NN	O	O
broader	NN	O	O
spectrum	NN	O	O
of	NN	O	O
mutations	NN	O	O
is	NN	O	O
selected	NN	O	O
in	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
rate	NN	O	O
is	NN	O	O
effectively	NN	O	O
higher	NN	O	O
and	NN	O	O
more	NN	O	O
tumors	NN	O	B-Disease
grow	NN	O	O
.	NN	O	O
.	NN	O	O

Mxi1	NN	O	O
mutations	NN	O	O
in	NN	O	O
human	NN	O	O
neurofibrosarcomas	NN	O	B-Disease
.	NN	O	O

Mxi1	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
Myc	NN	O	O
function	NN	O	O
and	NN	O	O
may	NN	O	O
therefore	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
.	NN	O	O

Little	NN	O	O
effort	NN	O	O
has	NN	O	O
yet	NN	O	O
been	NN	O	O
made	NN	O	O
to	NN	O	O
find	NN	O	O
alterations	NN	O	O
involving	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
human	NN	O	O
solid	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
screened	NN	O	O
31	NN	O	O
human	NN	O	O
gastric	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
7	NN	O	O
esophageal	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
85	NN	O	O
bone	NN	O	B-Disease
and	NN	O	I-Disease
soft	NN	O	I-Disease
tissue	NN	O	I-Disease
tumors	NN	O	I-Disease
of	NN	O	O
various	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
4	NN	O	O
neurofibrosarcomas	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
29	NN	O	O
human	NN	O	O
tumor	NN	O	B-Disease
cell	NN	O	O
lines	NN	O	O
consisting	NN	O	O
of	NN	O	O
12	NN	O	O
esophageal	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
7	NN	O	O
glioma	NN	O	B-Disease
/	NN	O	O
glioblastomas	NN	O	B-Disease
and	NN	O	O
10	NN	O	O
others	NN	O	O
for	NN	O	O
Mxi1	NN	O	O
mutations	NN	O	O
in	NN	O	O
exons	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
,	NN	O	O
4	NN	O	O
(	NN	O	O
HLH	NN	O	O
domain	NN	O	O
)	NN	O	O
,	NN	O	O
5	NN	O	O
and	NN	O	O
6	NN	O	O
.	NN	O	O

Polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
PCR	NN	O	O
-	NN	O	O
SSCP	NN	O	O
)	NN	O	O
and	NN	O	O
subsequent	NN	O	O
sequencing	NN	O	O
revealed	NN	O	O
three	NN	O	O
distinct	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
the	NN	O	O
intron	NN	O	O
-	NN	O	O
exon	NN	O	O
boundary	NN	O	O
upstream	NN	O	O
from	NN	O	O
exon	NN	O	O
6	NN	O	O
.	NN	O	O

We	NN	O	O
discovered	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
GCA	NN	O	O
to	NN	O	O
GTA	NN	O	O
(	NN	O	O
Ala	NN	O	O
54	NN	O	O
Val	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
exon	NN	O	O
2	NN	O	O
in	NN	O	O
a	NN	O	O
neurofibrosarcoma	NN	O	B-Disease
patient	NN	O	O
(	NN	O	O
case	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
AAA	NN	O	O
to	NN	O	O
CAA	NN	O	O
(	NN	O	O
Lys	NN	O	O
118	NN	O	O
Gln	NN	O	O
)	NN	O	O
and	NN	O	O
GAA	NN	O	O
to	NN	O	O
GGA	NN	O	O
(	NN	O	O
Glu	NN	O	O
154	NN	O	O
Gly	NN	O	O
)	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
of	NN	O	O
another	NN	O	O
neurofibrosarcoma	NN	O	B-Disease
patient	NN	O	O
(	NN	O	O
case	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
,	NN	O	O
GTG	NN	O	O
to	NN	O	O
GCG	NN	O	O
(	NN	O	O
Val	NN	O	O
179	NN	O	O
Ala	NN	O	O
)	NN	O	O
,	NN	O	O
GTT	NN	O	O
to	NN	O	O
GCT	NN	O	O
(	NN	O	O
Val	NN	O	O
181	NN	O	O
Ala	NN	O	O
)	NN	O	O
and	NN	O	O
TTC	NN	O	O
to	NN	O	O
CTC	NN	O	O
(	NN	O	O
Phe	NN	O	O
186	NN	O	O
Leu	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
third	NN	O	O
neurofibrosarcoma	NN	O	B-Disease
patient	NN	O	O
(	NN	O	O
case	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
case	NN	O	O
3	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
informative	NN	O	O
(	NN	O	O
TTC	NN	O	O
)	NN	O	O
3	NN	O	O
/	NN	O	O
(	NN	O	O
TTC	NN	O	O
)	NN	O	O
2	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
mutations	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
Mxi1	NN	O	O
gene	NN	O	O
in	NN	O	O
neurofibrosarcoma	NN	O	B-Disease
.	NN	O	O

Missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
functional	NN	O	O
domain	NN	O	O
of	NN	O	O
Mxi1	NN	O	O
in	NN	O	O
these	NN	O	O
cases	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
neurofibrosarcoma	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
hemochromatosis	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
Hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
homozygosity	NN	O	O
for	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
hemochromatosis	NN	O	B-Disease
(	NN	O	O
HFE	NN	O	O
)	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
6	NN	O	O
,	NN	O	O
elevated	NN	O	O
serum	NN	O	O
transferrin	NN	O	O
saturation	NN	O	O
,	NN	O	O
and	NN	O	O
excess	NN	O	B-Disease
iron	NN	O	I-Disease
deposits	NN	O	I-Disease
throughout	NN	O	O
the	NN	O	O
body	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
prevalence	NN	O	O
and	NN	O	O
clinical	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
HFE	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
conducted	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
study	NN	O	O
in	NN	O	O
Busselton	NN	O	O
,	NN	O	O
Australia	NN	O	O
.	NN	O	O

In	NN	O	O
1994	NN	O	O
,	NN	O	O
we	NN	O	O
obtained	NN	O	O
blood	NN	O	O
samples	NN	O	O
for	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
serum	NN	O	O
transferrin	NN	O	O
saturation	NN	O	O
and	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
and	NN	O	O
the	NN	O	O
H63D	NN	O	O
mutation	NN	O	O
(	NN	O	O
which	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
increased	NN	O	O
hepatic	NN	O	O
iron	NN	O	O
levels	NN	O	O
)	NN	O	O
in	NN	O	O
3011	NN	O	O
unrelated	NN	O	O
white	NN	O	O
adults	NN	O	O
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
all	NN	O	O
subjects	NN	O	O
who	NN	O	O
had	NN	O	O
persistently	NN	O	O
elevated	NN	O	O
transferrin	NN	O	O
-	NN	O	O
saturation	NN	O	O
values	NN	O	O
(	NN	O	O
45	NN	O	O
percent	NN	O	O
or	NN	O	O
higher	NN	O	O
)	NN	O	O
or	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
.	NN	O	O

We	NN	O	O
recommended	NN	O	O
liver	NN	O	O
biopsy	NN	O	O
for	NN	O	O
subjects	NN	O	O
with	NN	O	O
serum	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
of	NN	O	O
300	NN	O	O
ng	NN	O	O
per	NN	O	O
milliliter	NN	O	O
or	NN	O	O
higher	NN	O	O
.	NN	O	O

The	NN	O	O
subjects	NN	O	O
were	NN	O	O
followed	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
four	NN	O	O
years	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Sixteen	NN	O	O
of	NN	O	O
the	NN	O	O
subjects	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
5	NN	O	O
percent	NN	O	O
)	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
424	NN	O	O
(	NN	O	O
14	NN	O	O
.	NN	O	O
1	NN	O	O
percent	NN	O	O
)	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	O
transferrin	NN	O	O
saturation	NN	O	O
was	NN	O	O
45	NN	O	O
percent	NN	O	O
or	NN	O	O
higher	NN	O	O
in	NN	O	O
15	NN	O	O
of	NN	O	O
the	NN	O	O
16	NN	O	O
who	NN	O	O
were	NN	O	O
homozygous	NN	O	O
;	NN	O	O
in	NN	O	O
1	NN	O	O
subject	NN	O	O
it	NN	O	O
was	NN	O	O
43	NN	O	O
percent	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
homozygous	NN	O	O
subjects	NN	O	O
had	NN	O	O
previously	NN	O	O
been	NN	O	O
given	NN	O	O
a	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
hemochromatosis	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
12	NN	O	O
had	NN	O	O
not	NN	O	O
.	NN	O	O

Seven	NN	O	O
of	NN	O	O
these	NN	O	O
12	NN	O	O
patients	NN	O	O
had	NN	O	O
elevated	NN	O	O
serum	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
in	NN	O	O
1994	NN	O	O
;	NN	O	O
6	NN	O	O
of	NN	O	O
the	NN	O	O
7	NN	O	O
had	NN	O	O
further	NN	O	O
increases	NN	O	O
in	NN	O	O
1998	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
had	NN	O	O
a	NN	O	O
decrease	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
value	NN	O	O
remained	NN	O	O
elevated	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
four	NN	O	O
other	NN	O	O
homozygous	NN	O	O
patients	NN	O	O
remained	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
.	NN	O	O

Eleven	NN	O	O
of	NN	O	O
the	NN	O	O
16	NN	O	O
homozygous	NN	O	O
subjects	NN	O	O
underwent	NN	O	O
liver	NN	O	O
biopsy	NN	O	O
;	NN	O	O
3	NN	O	O
had	NN	O	O
hepatic	NN	O	B-Disease
fibrosis	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
who	NN	O	O
had	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
excessive	NN	O	B-Disease
alcohol	NN	O	I-Disease
consumption	NN	O	I-Disease
,	NN	O	O
had	NN	O	O
cirrhosis	NN	O	B-Disease
and	NN	O	O
mild	NN	O	O
microvesicular	NN	O	B-Disease
steatosis	NN	O	I-Disease
.	NN	O	O

Eight	NN	O	O
of	NN	O	O
the	NN	O	O
16	NN	O	O
homozygous	NN	O	O
subjects	NN	O	O
had	NN	O	O
clinical	NN	O	O
findings	NN	O	O
that	NN	O	O
were	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
hereditary	NN	O	B-Disease
hemochromatosis	NN	O	I-Disease
,	NN	O	O
such	NN	O	O
as	NN	O	O
hepatomegaly	NN	O	B-Disease
,	NN	O	O
skin	NN	O	B-Disease
pigmentation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
arthritis	NN	O	B-Disease
.	NN	O	O

CONCLUSIONS	NN	O	O
In	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
white	NN	O	O
adults	NN	O	O
of	NN	O	O
northern	NN	O	O
European	NN	O	O
ancestry	NN	O	O
,	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
percent	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
C282Y	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
HFE	NN	O	O
gene	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
half	NN	O	O
of	NN	O	O
those	NN	O	O
who	NN	O	O
were	NN	O	O
homozygous	NN	O	O
had	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
hemochromatosis	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
one	NN	O	O
quarter	NN	O	O
had	NN	O	O
serum	NN	O	O
ferritin	NN	O	O
levels	NN	O	O
that	NN	O	O
remained	NN	O	O
normal	NN	O	O
over	NN	O	O
a	NN	O	O
four	NN	O	O
-	NN	O	O
year	NN	O	O
period	NN	O	O
.	NN	O	O

Large	NN	O	O
heterozygous	NN	O	O
deletion	NN	O	O
masquerading	NN	O	O
as	NN	O	O
homozygous	NN	O	O
missense	NN	O	O
mutation	NN	O	O
:	NN	O	O
a	NN	O	O
pitfall	NN	O	O
in	NN	O	O
diagnostic	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
use	NN	O	O
of	NN	O	O
molecular	NN	O	O
analyses	NN	O	O
in	NN	O	O
recessive	NN	O	B-Disease
disorders	NN	O	I-Disease
relies	NN	O	O
on	NN	O	O
the	NN	O	O
exact	NN	O	O
characterization	NN	O	O
of	NN	O	O
both	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
patient	NN	O	O
.	NN	O	O

This	NN	O	O
can	NN	O	O
be	NN	O	O
problematic	NN	O	O
when	NN	O	O
only	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
is	NN	O	O
examined	NN	O	O
or	NN	O	O
when	NN	O	O
relevant	NN	O	O
DNA	NN	O	O
alterations	NN	O	O
are	NN	O	O
not	NN	O	O
recognized	NN	O	O
by	NN	O	O
standard	NN	O	O
methods	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
child	NN	O	O
in	NN	O	O
whom	NN	O	O
phenylketonuria	NN	O	B-Disease
was	NN	O	O
apparently	NN	O	O
caused	NN	O	O
by	NN	O	O
homozygosity	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
E390G	NN	O	O
in	NN	O	O
exon	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
clinical	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
was	NN	O	O
not	NN	O	O
quite	NN	O	O
as	NN	O	O
mild	NN	O	O
as	NN	O	O
expected	NN	O	O
,	NN	O	O
the	NN	O	O
mutation	NN	O	O
was	NN	O	O
not	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
father	NN	O	O
despite	NN	O	O
confirmed	NN	O	O
paternity	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
paternal	NN	O	O
allele	NN	O	O
showed	NN	O	O
a	NN	O	O
highly	NN	O	O
unusual	NN	O	O
pattern	NN	O	O
of	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

Presence	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
deletion	NN	O	O
involving	NN	O	O
exons	NN	O	O
9	NN	O	O
,	NN	O	O
10	NN	O	O
and	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
long	NN	O	O
-	NN	O	O
range	NN	O	O
PCR	NN	O	O
.	NN	O	O

Diagnostic	NN	O	O
DNA	NN	O	O
analyses	NN	O	O
should	NN	O	O
include	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
whole	NN	O	O
relevant	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
confirmation	NN	O	O
of	NN	O	O
carrier	NN	O	O
status	NN	O	O
in	NN	O	O
both	NN	O	O
parents	NN	O	O
.	NN	O	O
.	NN	O	O

Early	NN	O	O
onset	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
emerin	NN	O	O
gene	NN	O	O
deletion	NN	O	O
.	NN	O	O

A	NN	O	O
boy	NN	O	O
developed	NN	O	O
contractures	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
Achilles	NN	O	I-Disease
tendons	NN	O	I-Disease
at	NN	O	O
3	NN	O	O
years	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
postcervical	NN	O	O
muscles	NN	O	O
at	NN	O	O
7	NN	O	O
years	NN	O	O
,	NN	O	O
although	NN	O	O
neither	NN	O	O
contractures	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
elbows	NN	O	I-Disease
nor	NN	O	O
cardiac	NN	O	B-Disease
abnormality	NN	O	I-Disease
were	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
9	NN	O	O
years	NN	O	O
.	NN	O	O

Muscle	NN	O	O
computed	NN	O	O
tomography	NN	O	O
scanning	NN	O	O
revealed	NN	O	O
changes	NN	O	O
characteristic	NN	O	O
of	NN	O	O
muscle	NN	O	O
involvement	NN	O	O
.	NN	O	O

Emerin	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
biopsied	NN	O	O
muscle	NN	O	O
,	NN	O	O
and	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
and	NN	O	O
PCR	NN	O	O
-	NN	O	O
based	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
emerin	NN	O	O
gene	NN	O	O
demonstrated	NN	O	O
no	NN	O	O
amplification	NN	O	O
product	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
confirmed	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EDMD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
reinforce	NN	O	O
the	NN	O	O
necessity	NN	O	O
of	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
the	NN	O	O
membrane	NN	O	O
protein	NN	O	O
emerin	NN	O	O
in	NN	O	O
younger	NN	O	O
patients	NN	O	O
with	NN	O	O
possible	NN	O	O
EDMD	NN	O	B-Disease
before	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
typical	NN	O	O
symptoms	NN	O	O
,	NN	O	O
to	NN	O	O
avoid	NN	O	O
sudden	NN	O	B-Disease
cardiac	NN	O	I-Disease
death	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Duchenne	NN	O	B-Disease
/	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
:	NN	O	O
correlation	NN	O	O
of	NN	O	O
phenotype	NN	O	O
by	NN	O	O
electroretinography	NN	O	O
with	NN	O	O
sites	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
dark	NN	O	O
-	NN	O	O
adapted	NN	O	O
electroretinogram	NN	O	O
(	NN	O	O
ERG	NN	O	O
)	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
shows	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
in	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
amplitude	NN	O	O
.	NN	O	O

Genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
studies	NN	O	O
of	NN	O	O
mouse	NN	O	O
models	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
show	NN	O	O
position	NN	O	O
-	NN	O	O
specific	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
upon	NN	O	O
the	NN	O	O
phenotype	NN	O	O
mice	NN	O	O
with	NN	O	O
5	NN	O	O
defects	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
have	NN	O	O
normal	NN	O	O
ERGs	NN	O	O
,	NN	O	O
those	NN	O	O
with	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
region	NN	O	O
have	NN	O	O
a	NN	O	O
normal	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
amplitude	NN	O	O
associated	NN	O	O
with	NN	O	O
prolonged	NN	O	O
implicit	NN	O	O
times	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
and	NN	O	O
oscillatory	NN	O	O
potentials	NN	O	O
,	NN	O	O
and	NN	O	O
mice	NN	O	O
with	NN	O	O
3	NN	O	O
defects	NN	O	O
have	NN	O	O
a	NN	O	O
phenotype	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

The	NN	O	O
mouse	NN	O	O
studies	NN	O	O
suggest	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
carboxyl	NN	O	O
terminal	NN	O	O
dystrophin	NN	O	O
isoform	NN	O	O
,	NN	O	O
Dp260	NN	O	O
,	NN	O	O
in	NN	O	O
retinal	NN	O	O
electrophysiology	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
undertaken	NN	O	O
a	NN	O	O
systematic	NN	O	O
evaluation	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
patients	NN	O	O
through	NN	O	O
clinical	NN	O	O
examination	NN	O	O
and	NN	O	O
review	NN	O	O
of	NN	O	O
the	NN	O	O
literature	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
position	NN	O	O
-	NN	O	O
specific	NN	O	O
effects	NN	O	O
of	NN	O	O
mutations	NN	O	O
noted	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
man	NN	O	O
,	NN	O	O
a	NN	O	O
wider	NN	O	O
variation	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
defects	NN	O	I-Disease
correlate	NN	O	O
with	NN	O	O
reductions	NN	O	O
in	NN	O	O
the	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
amplitude	NN	O	O
.	NN	O	O

Individuals	NN	O	O
with	NN	O	O
normal	NN	O	O
ERGs	NN	O	O
have	NN	O	O
mutations	NN	O	O
predominantly	NN	O	O
located	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
transcript	NN	O	O
initiation	NN	O	O
site	NN	O	O
of	NN	O	O
Dp260	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
determinant	NN	O	O
in	NN	O	O
the	NN	O	O
ERG	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
phenotype	NN	O	O
is	NN	O	O
the	NN	O	O
mutation	NN	O	O
position	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
muscle	NN	O	B-Disease
disease	NN	O	I-Disease
severity	NN	O	O
.	NN	O	O

Forty	NN	O	O
-	NN	O	O
six	NN	O	O
per	NN	O	O
cent	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
mutations	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
Dp260	NN	O	O
transcript	NN	O	O
start	NN	O	O
site	NN	O	O
have	NN	O	O
abnormal	NN	O	O
ERGs	NN	O	O
,	NN	O	O
as	NN	O	O
opposed	NN	O	O
to	NN	O	O
94	NN	O	O
%	NN	O	O
with	NN	O	O
more	NN	O	O
distal	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
Dp260	NN	O	O
in	NN	O	O
normal	NN	O	O
retinal	NN	O	O
electrophysiology	NN	O	O
and	NN	O	O
may	NN	O	O
also	NN	O	O
reflect	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
dystrophin	NN	O	O
isoforms	NN	O	O
and	NN	O	O
their	NN	O	O
contributions	NN	O	O
to	NN	O	O
retinal	NN	O	O
signal	NN	O	O
transmission	NN	O	O
.	NN	O	O
.	NN	O	O

Cis	NN	O	O
and	NN	O	O
trans	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
culture	NN	O	O
model	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
causing	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
-	NN	O	O
untranslated	NN	O	O
region	NN	O	O
(	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
protein	NN	O	O
kinase	NN	O	O
gene	NN	O	O
(	NN	O	O
DMPK	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
pathogenesis	NN	O	O
remain	NN	O	O
unknown	NN	O	O
.	NN	O	O

Studies	NN	O	O
using	NN	O	O
DM	NN	O	B-Disease
patient	NN	O	O
materials	NN	O	O
have	NN	O	O
often	NN	O	O
produced	NN	O	O
confusing	NN	O	O
results	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
controlled	NN	O	O
environment	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
established	NN	O	O
a	NN	O	O
cell	NN	O	O
culture	NN	O	O
model	NN	O	O
system	NN	O	O
using	NN	O	O
C2C12	NN	O	O
mouse	NN	O	O
myoblasts	NN	O	O
.	NN	O	O

By	NN	O	O
expressing	NN	O	O
chimeric	NN	O	O
reporter	NN	O	O
constructs	NN	O	O
containing	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
fused	NN	O	O
to	NN	O	O
a	NN	O	O
human	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
both	NN	O	O
cis	NN	O	O
and	NN	O	O
trans	NN	O	O
effects	NN	O	O
that	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
mutant	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
,	NN	O	O
with	NN	O	O
as	NN	O	O
few	NN	O	O
as	NN	O	O
57	NN	O	O
CTGs	NN	O	O
,	NN	O	O
had	NN	O	O
a	NN	O	O
negative	NN	O	O
cis	NN	O	O
effect	NN	O	O
on	NN	O	O
protein	NN	O	O
expression	NN	O	O
and	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
aggregation	NN	O	O
of	NN	O	O
reporter	NN	O	O
transcripts	NN	O	O
into	NN	O	O
discrete	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
by	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
that	NN	O	O
an	NN	O	O
expanded	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
(	NN	O	O
n	NN	O	O
)	NN	O	O
tract	NN	O	O
alone	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
these	NN	O	O
cis	NN	O	O
effects	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
mRNA	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
mRNA	NN	O	O
with	NN	O	O
(	NN	O	O
CUG	NN	O	O
)	NN	O	O
(	NN	O	O
200	NN	O	O
)	NN	O	O
selectively	NN	O	O
inhibited	NN	O	O
myogenic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
C2C12	NN	O	O
myoblasts	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
and	NN	O	O
the	NN	O	O
Cre	NN	O	O
-	NN	O	O
loxP	NN	O	O
system	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
clearly	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
myoblast	NN	O	O
fusion	NN	O	O
defect	NN	O	O
could	NN	O	O
be	NN	O	O
rescued	NN	O	O
by	NN	O	O
eliminating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
transcript	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
deletion	NN	O	O
events	NN	O	O
mapped	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
to	NN	O	O
the	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
(	NN	O	O
n	NN	O	O
)	NN	O	O
expansion	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
the	NN	O	O
3	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
DMPK	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
acts	NN	O	O
in	NN	O	O
cis	NN	O	O
to	NN	O	O
reduce	NN	O	O
protein	NN	O	O
production	NN	O	O
(	NN	O	O
consistent	NN	O	O
with	NN	O	O
DMPK	NN	O	B-Disease
haploinsufficiency	NN	O	I-Disease
)	NN	O	O
and	NN	O	O
in	NN	O	O
trans	NN	O	O
as	NN	O	O
a	NN	O	O
riboregulator	NN	O	O
to	NN	O	O
inhibit	NN	O	O
myogenesis	NN	O	O
.	NN	O	O
.	NN	O	O

Coats	NN	O	B-Disease
'	NN	O	I-Disease
disease	NN	O	I-Disease
of	NN	O	O
the	NN	O	O
retina	NN	O	O
(	NN	O	O
unilateral	NN	O	B-Disease
retinal	NN	O	I-Disease
telangiectasis	NN	O	I-Disease
)	NN	O	O
caused	NN	O	O
by	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
:	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
norrin	NN	O	O
in	NN	O	O
retinal	NN	O	O
angiogenesis	NN	O	O
.	NN	O	O

Coats	NN	O	B-Disease
disease	NN	O	I-Disease
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
abnormal	NN	O	B-Disease
retinal	NN	O	I-Disease
vascular	NN	O	I-Disease
development	NN	O	I-Disease
(	NN	O	O
so	NN	O	O
-	NN	O	O
called	NN	O	O
retinal	NN	O	B-Disease
telangiectasis	NN	O	I-Disease
)	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
massive	NN	O	O
intraretinal	NN	O	B-Disease
and	NN	O	I-Disease
subretinal	NN	O	I-Disease
lipid	NN	O	I-Disease
accumulation	NN	O	I-Disease
(	NN	O	O
exudative	NN	O	B-Disease
retinal	NN	O	I-Disease
detachment	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
classical	NN	O	O
form	NN	O	O
of	NN	O	O
Coats	NN	O	B-Disease
disease	NN	O	I-Disease
is	NN	O	O
almost	NN	O	O
invariably	NN	O	O
isolated	NN	O	O
,	NN	O	O
unilateral	NN	O	O
and	NN	O	O
seen	NN	O	O
in	NN	O	O
males	NN	O	O
.	NN	O	O

A	NN	O	O
female	NN	O	O
with	NN	O	O
a	NN	O	O
unilateral	NN	O	O
variant	NN	O	O
of	NN	O	O
Coats	NN	O	B-Disease
disease	NN	O	I-Disease
gave	NN	O	O
birth	NN	O	O
to	NN	O	O
a	NN	O	O
son	NN	O	O
affected	NN	O	O
by	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Both	NN	O	O
carried	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
within	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
Xp11	NN	O	O
.	NN	O	O

2	NN	O	O
2	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
retinas	NN	O	O
of	NN	O	O
nine	NN	O	O
enucleated	NN	O	O
eyes	NN	O	O
from	NN	O	O
males	NN	O	O
with	NN	O	O
Coats	NN	O	B-Disease
disease	NN	O	I-Disease
demonstrated	NN	O	O
in	NN	O	O
one	NN	O	O
a	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
present	NN	O	O
within	NN	O	O
non	NN	O	O
-	NN	O	O
retinal	NN	O	O
tissue	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
Coats	NN	O	B-Disease
telangiectasis	NN	O	I-Disease
is	NN	O	O
secondary	NN	O	O
to	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
norrin	NN	O	I-Disease
(	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
)	NN	O	O
within	NN	O	O
the	NN	O	O
developing	NN	O	O
retina	NN	O	O
.	NN	O	O

This	NN	O	O
supports	NN	O	O
recent	NN	O	O
observations	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
normal	NN	O	O
retinal	NN	O	O
vasculogenesis	NN	O	O
.	NN	O	O

Hereditary	NN	O	O
TP53	NN	O	O
codon	NN	O	O
292	NN	O	O
and	NN	O	O
somatic	NN	O	O
P16INK4A	NN	O	O
codon	NN	O	O
94	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
Li	NN	O	B-Disease
-	NN	O	I-Disease
Fraumeni	NN	O	I-Disease
syndrome	NN	O	I-Disease
family	NN	O	O
.	NN	O	O

Li	NN	O	B-Disease
-	NN	O	I-Disease
Fraumeni	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
disorder	NN	O	I-Disease
that	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
in	NN	O	O
childhood	NN	O	O
and	NN	O	O
adult	NN	O	O
cases	NN	O	O
.	NN	O	O

Although	NN	O	O
hereditary	NN	O	O
TP53	NN	O	O
mutation	NN	O	O
is	NN	O	O
very	NN	O	O
rare	NN	O	O
in	NN	O	O
different	NN	O	O
human	NN	O	O
cancers	NN	O	B-Disease
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
frequently	NN	O	O
reported	NN	O	O
in	NN	O	O
Li	NN	O	B-Disease
-	NN	O	I-Disease
Fraumeni	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
hereditary	NN	O	O
mutations	NN	O	O
of	NN	O	O
TP57KIP2	NN	O	O
,	NN	O	O
P15INK4B	NN	O	O
,	NN	O	O
and	NN	O	O
P16INK4A	NN	O	O
,	NN	O	O
which	NN	O	O
affect	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
similar	NN	O	O
to	NN	O	O
TP53	NN	O	O
,	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
some	NN	O	O
types	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
a	NN	O	O
Turkish	NN	O	O
family	NN	O	O
with	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Li	NN	O	B-Disease
-	NN	O	I-Disease
Fraumeni	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
mutation	NN	O	O
pattern	NN	O	O
of	NN	O	O
TP53	NN	O	O
,	NN	O	O
P57KIP2	NN	O	O
,	NN	O	O
P15INK4B	NN	O	O
,	NN	O	O
and	NN	O	O
P16INK4A	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
,	NN	O	O
and	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
(	NN	O	O
homo	NN	O	O
/	NN	O	O
hemizygous	NN	O	O
deletion	NN	O	O
)	NN	O	O
pattern	NN	O	O
of	NN	O	O
TP53	NN	O	O
and	NN	O	O
P15INK4B	NN	O	O
/	NN	O	O
P16INK4A	NN	O	O
in	NN	O	O
two	NN	O	O
tumor	NN	O	B-Disease
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
propositus	NN	O	O
had	NN	O	O
a	NN	O	O
seminoma	NN	O	B-Disease
,	NN	O	O
his	NN	O	O
daughter	NN	O	O
a	NN	O	O
medulloblastoma	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
one	NN	O	O
of	NN	O	O
his	NN	O	O
healthy	NN	O	O
cousins	NN	O	O
,	NN	O	O
a	NN	O	O
TP53	NN	O	O
codon	NN	O	O
292	NN	O	O
missense	NN	O	O
point	NN	O	O
mutation	NN	O	O
(	NN	O	O
AAA	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
ATA	NN	O	O
;	NN	O	O
Lys	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Ile	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
cells	NN	O	O
.	NN	O	O

Tumor	NN	O	O
tissue	NN	O	O
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
propositus	NN	O	O
with	NN	O	O
the	NN	O	O
seminoma	NN	O	B-Disease
revealed	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
in	NN	O	O
the	NN	O	O
TP53	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
analyses	NN	O	O
of	NN	O	O
tumor	NN	O	B-Disease
tissues	NN	O	O
from	NN	O	O
the	NN	O	O
propositus	NN	O	O
and	NN	O	O
his	NN	O	O
daughter	NN	O	O
,	NN	O	O
a	NN	O	O
P16INK4A	NN	O	O
codon	NN	O	O
94	NN	O	O
missense	NN	O	O
point	NN	O	O
mutation	NN	O	O
(	NN	O	O
GCG	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
GAG	NN	O	O
;	NN	O	O
Ala	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Glu	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
hereditary	NN	O	O
TP53	NN	O	O
mutation	NN	O	O
.	NN	O	O

P16INK4A	NN	O	O
codon	NN	O	O
94	NN	O	O
mutation	NN	O	O
observed	NN	O	O
in	NN	O	O
our	NN	O	O
family	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
Li	NN	O	B-Disease
-	NN	O	I-Disease
Fraumeni	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

No	NN	O	O
other	NN	O	O
gene	NN	O	O
alteration	NN	O	O
in	NN	O	O
TP53	NN	O	O
,	NN	O	O
P57KIP2	NN	O	O
,	NN	O	O
P15INK4B	NN	O	O
,	NN	O	O
and	NN	O	O
P16INK4A	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Existence	NN	O	O
of	NN	O	O
the	NN	O	O
P16INK4A	NN	O	O
mutation	NN	O	O
and	NN	O	O
the	NN	O	O
hereditary	NN	O	O
TP53	NN	O	O
mutation	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
in	NN	O	O
the	NN	O	O
TP53	NN	O	O
gene	NN	O	O
(	NN	O	O
seminoma	NN	O	B-Disease
/	NN	O	O
medulloblastoma	NN	O	B-Disease
)	NN	O	O
may	NN	O	O
be	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
involved	NN	O	O
in	NN	O	O
tumorogenesis	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
gene	NN	O	O
alterations	NN	O	O
in	NN	O	O
TP53	NN	O	O
and	NN	O	O
P16INK4A	NN	O	O
genes	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
tumor	NN	O	B-Disease
markers	NN	O	O
in	NN	O	O
our	NN	O	O
family	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
sodium	NN	O	O
/	NN	O	O
iodide	NN	O	O
symporter	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
largest	NN	O	O
family	NN	O	O
with	NN	O	O
iodide	NN	O	B-Disease
transport	NN	O	I-Disease
defect	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
nine	NN	O	O
children	NN	O	O
with	NN	O	O
an	NN	O	O
autosomally	NN	O	O
recessive	NN	O	O
form	NN	O	O
of	NN	O	O
congenital	NN	O	B-Disease
hypothyroidism	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
an	NN	O	O
iodide	NN	O	B-Disease
transport	NN	O	I-Disease
defect	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
large	NN	O	O
Hutterite	NN	O	O
family	NN	O	O
with	NN	O	O
extensive	NN	O	O
consanguinity	NN	O	O
living	NN	O	O
in	NN	O	O
central	NN	O	O
Canada	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
original	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
diagnosed	NN	O	O
congenital	NN	O	B-Disease
hypothyroidism	NN	O	I-Disease
by	NN	O	O
newborn	NN	O	O
TSH	NN	O	O
screening	NN	O	O
in	NN	O	O
9	NN	O	O
additional	NN	O	O
children	NN	O	O
from	NN	O	O
the	NN	O	O
family	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
PCR	NN	O	O
products	NN	O	O
of	NN	O	O
each	NN	O	O
NIS	NN	O	O
(	NN	O	O
sodium	NN	O	O
/	NN	O	O
iodide	NN	O	O
symporter	NN	O	O
)	NN	O	O
gene	NN	O	O
exon	NN	O	O
with	NN	O	O
flanking	NN	O	O
introns	NN	O	O
amplified	NN	O	O
from	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
extracted	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
NIS	NN	O	O
gene	NN	O	O
mutation	NN	O	O
,	NN	O	O
G395R	NN	O	O
(	NN	O	O
Gly395	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Arg	NN	O	O
;	NN	O	O
GGA	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
AGA	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
10	NN	O	O
patients	NN	O	O
examined	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
parents	NN	O	O
tested	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
homozygous	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
was	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
10th	NN	O	O
transmembrane	NN	O	O
helix	NN	O	O
.	NN	O	O

Expression	NN	O	O
experiments	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
NIS	NN	O	O
complimentary	NN	O	O
DNA	NN	O	O
into	NN	O	O
COS	NN	O	O
-	NN	O	O
7	NN	O	O
cells	NN	O	O
showed	NN	O	O
no	NN	O	O
perchlorate	NN	O	O
-	NN	O	O
sensitive	NN	O	O
iodide	NN	O	O
uptake	NN	O	O
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
direct	NN	O	O
cause	NN	O	O
of	NN	O	O
the	NN	O	O
iodide	NN	O	B-Disease
transport	NN	O	I-Disease
defect	NN	O	I-Disease
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

A	NN	O	O
patient	NN	O	O
who	NN	O	O
showed	NN	O	O
an	NN	O	O
intermediate	NN	O	O
saliva	NN	O	O
/	NN	O	O
serum	NN	O	O
technetium	NN	O	O
ratio	NN	O	O
(	NN	O	O
14	NN	O	O
.	NN	O	O
0	NN	O	O
;	NN	O	O
normal	NN	O	O
,	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
20	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
considered	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
partial	NN	O	O
or	NN	O	O
less	NN	O	O
severe	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
previous	NN	O	O
report	NN	O	O
(	NN	O	O
IX	NN	O	O
-	NN	O	O
24	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
a	NN	O	O
NIS	NN	O	O
gene	NN	O	O
mutation	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
now	NN	O	O
possible	NN	O	O
to	NN	O	O
use	NN	O	O
gene	NN	O	O
diagnostics	NN	O	O
of	NN	O	O
this	NN	O	O
unique	NN	O	O
NIS	NN	O	O
mutation	NN	O	O
to	NN	O	O
identify	NN	O	O
patients	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
hypothyroidism	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
an	NN	O	O
iodide	NN	O	B-Disease
transport	NN	O	I-Disease
defect	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
family	NN	O	O
and	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
carrier	NN	O	O
state	NN	O	O
of	NN	O	O
potential	NN	O	O
parents	NN	O	O
for	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
and	NN	O	O
arranging	NN	O	O
rapid	NN	O	O
and	NN	O	O
early	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
their	NN	O	O
infants	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
:	NN	O	O
early	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
an	NN	O	O
asymptomatic	NN	O	O
carrier	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
(	NN	O	O
FNDI	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
hormone	NN	O	I-Disease
arginine	NN	O	I-Disease
vasopressin	NN	O	I-Disease
(	NN	O	O
AVP	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
transmitted	NN	O	O
as	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
trait	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
AVP	NN	O	O
-	NN	O	O
neurophysin	NN	O	O
II	NN	O	O
(	NN	O	O
AVP	NN	O	O
-	NN	O	O
NPII	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
Spanish	NN	O	O
kindred	NN	O	O
.	NN	O	O

Studies	NN	O	O
were	NN	O	O
performed	NN	O	O
on	NN	O	O
seven	NN	O	O
members	NN	O	O
(	NN	O	O
four	NN	O	O
clinically	NN	O	O
affected	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
.	NN	O	O

Patients	NN	O	O
were	NN	O	O
diagnosed	NN	O	O
at	NN	O	O
the	NN	O	O
Hospital	NN	O	O
Universitario	NN	O	O
Gregorio	NN	O	O
Maranon	NN	O	O
(	NN	O	O
Madrid	NN	O	O
,	NN	O	O
Spain	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
AVP	NN	O	O
-	NN	O	O
NPII	NN	O	O
gene	NN	O	O
of	NN	O	O
all	NN	O	O
family	NN	O	O
members	NN	O	O
was	NN	O	O
amplified	NN	O	O
by	NN	O	O
PCR	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

All	NN	O	O
affected	NN	O	O
individuals	NN	O	O
presented	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
G1757	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
)	NN	O	O
that	NN	O	O
replaces	NN	O	O
glycine	NN	O	O
at	NN	O	O
position	NN	O	O
23	NN	O	O
with	NN	O	O
arginine	NN	O	O
within	NN	O	O
the	NN	O	O
NPII	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
substitution	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
restriction	NN	O	O
endonuclease	NN	O	O
analysis	NN	O	O
and	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
heterozygosis	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
asymptomatic	NN	O	O
relatives	NN	O	O
(	NN	O	O
a	NN	O	O
girl	NN	O	O
8	NN	O	O
months	NN	O	O
old	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
study	NN	O	O
)	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
carrier	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
mutation	NN	O	O
and	NN	O	O
developed	NN	O	O
the	NN	O	O
disease	NN	O	O
3	NN	O	O
months	NN	O	O
later	NN	O	O
.	NN	O	O

The	NN	O	O
alteration	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
as	NN	O	O
all	NN	O	O
individuals	NN	O	O
harboring	NN	O	O
the	NN	O	O
mutation	NN	O	O
had	NN	O	O
been	NN	O	O
previously	NN	O	O
diagnosed	NN	O	O
or	NN	O	O
have	NN	O	O
eventually	NN	O	O
developed	NN	O	O
FNDI	NN	O	B-Disease
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
defect	NN	O	O
underlying	NN	O	O
FNDI	NN	O	B-Disease
in	NN	O	O
affected	NN	O	O
families	NN	O	O
is	NN	O	O
a	NN	O	O
powerful	NN	O	O
tool	NN	O	O
for	NN	O	O
early	NN	O	O
asymptomatic	NN	O	O
diagnosis	NN	O	O
in	NN	O	O
infants	NN	O	O
.	NN	O	O
.	NN	O	O

Deficit	NN	O	O
of	NN	O	O
in	NN	O	O
vivo	NN	O	O
mitochondrial	NN	O	O
ATP	NN	O	O
production	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
of	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
ataxias	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
degenerative	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
characterized	NN	O	O
clinically	NN	O	O
by	NN	O	O
onset	NN	O	O
before	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
25	NN	O	O
of	NN	O	O
progressive	NN	O	B-Disease
gait	NN	O	I-Disease
and	NN	O	I-Disease
limb	NN	O	I-Disease
ataxia	NN	O	I-Disease
,	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
deep	NN	O	I-Disease
tendon	NN	O	I-Disease
reflexes	NN	O	I-Disease
,	NN	O	O
extensor	NN	O	B-Disease
plantar	NN	O	I-Disease
responses	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
loss	NN	O	B-Disease
of	NN	O	I-Disease
position	NN	O	I-Disease
and	NN	O	I-Disease
vibration	NN	O	I-Disease
sense	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
lower	NN	O	O
limbs	NN	O	O
.	NN	O	O

FRDA	NN	O	B-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
GAA	NN	O	O
triplet	NN	O	O
expansion	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
intron	NN	O	O
of	NN	O	O
the	NN	O	O
FRDA	NN	O	B-Disease
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
9q13	NN	O	O
in	NN	O	O
97	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
FRDA	NN	O	B-Disease
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
widely	NN	O	O
expressed	NN	O	O
210	NN	O	O
-	NN	O	O
aa	NN	O	O
protein	NN	O	O
,	NN	O	O
frataxin	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
mitochondria	NN	O	O
and	NN	O	O
is	NN	O	O
severely	NN	O	O
reduced	NN	O	O
in	NN	O	O
FRDA	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Frataxin	NN	O	O
function	NN	O	O
is	NN	O	O
still	NN	O	O
unknown	NN	O	O
but	NN	O	O
the	NN	O	O
knockout	NN	O	O
of	NN	O	O
the	NN	O	O
yeast	NN	O	O
frataxin	NN	O	O
homologue	NN	O	O
gene	NN	O	O
(	NN	O	O
YFH1	NN	O	O
)	NN	O	O
showed	NN	O	O
a	NN	O	O
severe	NN	O	O
defect	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
respiration	NN	O	O
and	NN	O	O
loss	NN	O	O
of	NN	O	O
mtDNA	NN	O	O
associated	NN	O	O
with	NN	O	O
elevated	NN	O	O
intramitochondrial	NN	O	O
iron	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
in	NN	O	O
vivo	NN	O	O
evidence	NN	O	O
of	NN	O	O
impaired	NN	O	O
mitochondrial	NN	O	O
respiration	NN	O	O
in	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
of	NN	O	O
FRDA	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Using	NN	O	O
phosphorus	NN	O	O
magnetic	NN	O	O
resonance	NN	O	O
spectroscopy	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
maximum	NN	O	O
rate	NN	O	O
of	NN	O	O
muscle	NN	O	O
mitochondrial	NN	O	O
ATP	NN	O	O
production	NN	O	O
(	NN	O	O
V	NN	O	O
(	NN	O	O
max	NN	O	O
)	NN	O	O
)	NN	O	O
below	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
in	NN	O	O
all	NN	O	O
12	NN	O	O
FRDA	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
a	NN	O	O
strong	NN	O	O
negative	NN	O	O
correlation	NN	O	O
between	NN	O	O
mitochondrial	NN	O	O
V	NN	O	O
(	NN	O	O
max	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GAA	NN	O	O
repeats	NN	O	O
in	NN	O	O
the	NN	O	O
smaller	NN	O	O
allele	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
FRDA	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
nuclear	NN	O	O
-	NN	O	O
encoded	NN	O	O
mitochondrial	NN	O	B-Disease
disorder	NN	O	I-Disease
affecting	NN	O	O
oxidative	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
give	NN	O	O
a	NN	O	O
rationale	NN	O	O
for	NN	O	O
treatments	NN	O	O
aimed	NN	O	O
to	NN	O	O
improve	NN	O	O
mitochondrial	NN	O	O
function	NN	O	O
in	NN	O	O
this	NN	O	O
condition	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
R21X	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
-	NN	O	O
type	NN	O	O
arginase	NN	O	O
gene	NN	O	O
(	NN	O	O
ARG1	NN	O	O
)	NN	O	O
in	NN	O	O
four	NN	O	O
Portuguese	NN	O	O
patients	NN	O	O
with	NN	O	O
argininemia	NN	O	B-Disease
.	NN	O	O

Argininemia	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
autossomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
deficiency	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
cytosolic	NN	O	I-Disease
liver	NN	O	I-Disease
-	NN	O	I-Disease
type	NN	O	I-Disease
arginase	NN	O	I-Disease
enzyme	NN	O	I-Disease
(	NN	O	O
L	NN	O	O
-	NN	O	O
arginine	NN	O	O
urea	NN	O	O
-	NN	O	O
hydrolase	NN	O	O
;	NN	O	O
E	NN	O	O
.	NN	O	O
C	NN	O	O
.	NN	O	O
3	NN	O	O
.	NN	O	O
5	NN	O	O
.	NN	O	O
3	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
argininemia	NN	O	B-Disease
in	NN	O	O
four	NN	O	O
unrelated	NN	O	O
Portuguese	NN	O	O
patients	NN	O	O
(	NN	O	O
two	NN	O	O
from	NN	O	O
northern	NN	O	O
Portugal	NN	O	O
and	NN	O	O
two	NN	O	O
from	NN	O	O
Madeira	NN	O	O
Island	NN	O	O
)	NN	O	O
we	NN	O	O
performed	NN	O	O
a	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
exons	NN	O	O
and	NN	O	O
exon	NN	O	O
/	NN	O	O
intron	NN	O	O
boundaries	NN	O	O
of	NN	O	O
the	NN	O	O
liver	NN	O	O
-	NN	O	O
type	NN	O	O
arginase	NN	O	O
gene	NN	O	O
(	NN	O	O
ARG1	NN	O	O
)	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
C	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
codon	NN	O	O
21	NN	O	O
(	NN	O	O
exon	NN	O	O
2	NN	O	O
)	NN	O	O
substituting	NN	O	O
arginine	NN	O	O
for	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
(	NN	O	O
R21X	NN	O	O
CGA	NN	O	O
to	NN	O	O
TGA	NN	O	O
)	NN	O	O
and	NN	O	O
generating	NN	O	O
a	NN	O	O
NlaIII	NN	O	O
restriction	NN	O	O
site	NN	O	O
.	NN	O	O

Restriction	NN	O	O
digestion	NN	O	O
following	NN	O	O
PCR	NN	O	O
amplification	NN	O	O
of	NN	O	O
ARG1	NN	O	O
exon	NN	O	O
2	NN	O	O
confirmed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
sterol	NN	O	O
27	NN	O	O
-	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
(	NN	O	O
CYP27	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
truncation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
expressed	NN	O	O
in	NN	O	O
leucocytes	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
family	NN	O	O
with	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
.	NN	O	O

OBJECTIVES	NN	O	O
A	NN	O	O
Japanese	NN	O	O
family	NN	O	O
with	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
investigated	NN	O	O
for	NN	O	O
a	NN	O	O
sequence	NN	O	O
alteration	NN	O	O
in	NN	O	O
the	NN	O	O
sterol	NN	O	O
27	NN	O	O
-	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
(	NN	O	O
CYP27	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
CYP27	NN	O	O
has	NN	O	O
been	NN	O	O
mostly	NN	O	O
explored	NN	O	O
using	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
prompting	NN	O	O
the	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
transcripts	NN	O	O
from	NN	O	O
blood	NN	O	O
leucocytes	NN	O	O
as	NN	O	O
a	NN	O	O
simple	NN	O	O
and	NN	O	O
rapid	NN	O	O
technique	NN	O	O
.	NN	O	O

METHODS	NN	O	O
An	NN	O	O
alteration	NN	O	O
in	NN	O	O
CYP27	NN	O	O
of	NN	O	O
the	NN	O	O
proband	NN	O	O
was	NN	O	O
searched	NN	O	O
for	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
PCR	NN	O	O
-	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
and	NN	O	O
subsequent	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Samples	NN	O	O
of	NN	O	O
RNA	NN	O	O
were	NN	O	O
subjected	NN	O	O
to	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
PCR	NN	O	O
(	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
proband	NN	O	O
was	NN	O	O
amplified	NN	O	O
with	NN	O	O
nested	NN	O	O
primers	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
A	NN	O	O
homozygous	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
intron	NN	O	O
7	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

In	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
analysis	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
truncated	NN	O	O
transcript	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
normal	NN	O	O
and	NN	O	O
truncated	NN	O	O
transcripts	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
siblings	NN	O	O
.	NN	O	O

The	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
cDNA	NN	O	O
fragment	NN	O	O
disclosed	NN	O	O
a	NN	O	O
direct	NN	O	O
conjuction	NN	O	O
of	NN	O	O
exon	NN	O	O
6	NN	O	O
and	NN	O	O
exon	NN	O	O
8	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
The	NN	O	O
mutation	NN	O	O
at	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
and	NN	O	O
the	NN	O	O
truncation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
were	NN	O	O
identical	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
a	NN	O	O
recently	NN	O	O
reported	NN	O	O
Italian	NN	O	O
patient	NN	O	O
,	NN	O	O
although	NN	O	O
different	NN	O	O
in	NN	O	O
symptomatology	NN	O	O
.	NN	O	O

The	NN	O	O
application	NN	O	O
of	NN	O	O
blood	NN	O	O
leucocytes	NN	O	O
can	NN	O	O
be	NN	O	O
a	NN	O	O
simple	NN	O	O
technique	NN	O	O
on	NN	O	O
analysing	NN	O	O
a	NN	O	O
constructive	NN	O	O
abnormality	NN	O	B-Disease
of	NN	O	I-Disease
CYP27	NN	O	I-Disease
mRNA	NN	O	O
.	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
19	NN	O	O
Wolfram	NN	O	B-Disease
syndrome	NN	O	I-Disease
kindreds	NN	O	O
demonstrating	NN	O	O
a	NN	O	O
wide	NN	O	O
spectrum	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
WFS1	NN	O	O
.	NN	O	O

Wolfram	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
neurodegenerative	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
juvenile	NN	O	B-Disease
-	NN	O	I-Disease
onset	NN	O	I-Disease
diabetes	NN	O	I-Disease
mellitus	NN	O	I-Disease
and	NN	O	O
progressive	NN	O	O
optic	NN	O	B-Disease
atrophy	NN	O	I-Disease
.	NN	O	O

mtDNA	NN	O	O
deletions	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
gene	NN	O	O
(	NN	O	O
WFS1	NN	O	O
)	NN	O	O
recently	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
on	NN	O	O
chromosome	NN	O	O
4p16	NN	O	O
,	NN	O	O
encoding	NN	O	O
a	NN	O	O
predicted	NN	O	O
890	NN	O	O
amino	NN	O	O
acid	NN	O	O
transmembrane	NN	O	O
protein	NN	O	O
.	NN	O	O

Direct	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
was	NN	O	O
done	NN	O	O
to	NN	O	O
screen	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
WFS1	NN	O	O
gene	NN	O	O
in	NN	O	O
30	NN	O	O
patients	NN	O	O
from	NN	O	O
19	NN	O	O
British	NN	O	O
kindreds	NN	O	O
with	NN	O	O
Wolfram	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

DNA	NN	O	O
was	NN	O	O
also	NN	O	O
screened	NN	O	O
for	NN	O	O
structural	NN	O	O
rearrangements	NN	O	O
(	NN	O	O
deletions	NN	O	O
and	NN	O	O
duplications	NN	O	O
)	NN	O	O
and	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
mtDNA	NN	O	O
.	NN	O	O

No	NN	O	O
pathogenic	NN	O	O
mtDNA	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
our	NN	O	O
cohort	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
24	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
WFS1	NN	O	O
gene	NN	O	O
8	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
8	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
3	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletions	NN	O	O
,	NN	O	O
1	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
insertion	NN	O	O
,	NN	O	O
and	NN	O	O
4	NN	O	O
frameshift	NN	O	O
mutations	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
23	NN	O	O
were	NN	O	O
novel	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
most	NN	O	O
occurred	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
were	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
two	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
common	NN	O	O
founder	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
were	NN	O	O
also	NN	O	O
analyzed	NN	O	O
for	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
relationships	NN	O	O
.	NN	O	O

Although	NN	O	O
some	NN	O	O
interesting	NN	O	O
cases	NN	O	O
were	NN	O	O
noted	NN	O	O
,	NN	O	O
consideration	NN	O	O
of	NN	O	O
the	NN	O	O
small	NN	O	O
sample	NN	O	O
size	NN	O	O
and	NN	O	O
frequency	NN	O	O
of	NN	O	O
each	NN	O	O
mutation	NN	O	O
indicated	NN	O	O
no	NN	O	O
clear	NN	O	O
-	NN	O	O
cut	NN	O	O
correlations	NN	O	O
between	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
observed	NN	O	O
mutations	NN	O	O
and	NN	O	O
disease	NN	O	O
severity	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
obvious	NN	O	O
mutation	NN	O	O
hot	NN	O	O
spots	NN	O	O
or	NN	O	O
clusters	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
molecular	NN	O	O
screening	NN	O	O
for	NN	O	O
Wolfram	NN	O	B-Disease
syndrome	NN	O	I-Disease
in	NN	O	O
affected	NN	O	O
families	NN	O	O
and	NN	O	O
for	NN	O	O
Wolfram	NN	O	B-Disease
syndrome	NN	O	I-Disease
-	NN	O	O
carrier	NN	O	O
status	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
psychiatric	NN	O	B-Disease
disorders	NN	O	I-Disease
or	NN	O	O
diabetes	NN	O	B-Disease
mellitus	NN	O	I-Disease
will	NN	O	O
require	NN	O	O
complete	NN	O	O
analysis	NN	O	O
of	NN	O	O
exon	NN	O	O
8	NN	O	O
and	NN	O	O
upstream	NN	O	O
exons	NN	O	O
.	NN	O	O
.	NN	O	O

Late	NN	O	O
-	NN	O	O
onset	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
:	NN	O	O
a	NN	O	O
subset	NN	O	O
with	NN	O	O
distinct	NN	O	O
clinical	NN	O	O
,	NN	O	O
demographic	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
characteristics	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
prevalence	NN	O	O
and	NN	O	O
characterize	NN	O	O
demographic	NN	O	O
,	NN	O	O
clinical	NN	O	O
,	NN	O	O
and	NN	O	O
genetic	NN	O	O
features	NN	O	O
of	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
of	NN	O	O
late	NN	O	O
onset	NN	O	O
,	NN	O	O
all	NN	O	O
patients	NN	O	O
experiencing	NN	O	O
their	NN	O	O
first	NN	O	O
FMF	NN	O	B-Disease
attack	NN	O	O
at	NN	O	O
age	NN	O	O
40	NN	O	O
years	NN	O	O
or	NN	O	O
more	NN	O	O
were	NN	O	O
identified	NN	O	O
using	NN	O	O
the	NN	O	O
computerized	NN	O	O
registry	NN	O	O
of	NN	O	O
our	NN	O	O
FMF	NN	O	B-Disease
clinic	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
thoroughly	NN	O	O
interviewed	NN	O	O
and	NN	O	O
examined	NN	O	O
.	NN	O	O

The	NN	O	O
control	NN	O	O
group	NN	O	O
consisted	NN	O	O
of	NN	O	O
40	NN	O	O
consecutive	NN	O	O
FMF	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
who	NN	O	O
arrived	NN	O	O
at	NN	O	O
the	NN	O	O
FMF	NN	O	B-Disease
clinic	NN	O	O
for	NN	O	O
their	NN	O	O
regular	NN	O	O
follow	NN	O	O
-	NN	O	O
up	NN	O	O
visit	NN	O	O
and	NN	O	O
were	NN	O	O
40	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
or	NN	O	O
older	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
the	NN	O	O
examination	NN	O	O
.	NN	O	O

The	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
in	NN	O	O
patients	NN	O	O
and	NN	O	O
controls	NN	O	O
was	NN	O	O
determined	NN	O	O
using	NN	O	O
a	NN	O	O
modified	NN	O	O
score	NN	O	O
,	NN	O	O
developed	NN	O	O
previously	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
in	NN	O	O
the	NN	O	O
FMF	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
commercial	NN	O	O
kit	NN	O	O
.	NN	O	O

Only	NN	O	O
20	NN	O	O
of	NN	O	O
4000	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
patients	NN	O	O
had	NN	O	O
late	NN	O	O
-	NN	O	O
onset	NN	O	O
FMF	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
patients	NN	O	O
were	NN	O	O
mostly	NN	O	O
men	NN	O	O
,	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
North	NN	O	O
African	NN	O	O
origin	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

05	NN	O	O
compared	NN	O	O
to	NN	O	O
controls	NN	O	O
.	NN	O	O

All	NN	O	O
had	NN	O	O
abdominal	NN	O	O
attacks	NN	O	O
and	NN	O	O
in	NN	O	O
most	NN	O	O
these	NN	O	O
were	NN	O	O
the	NN	O	O
only	NN	O	O
manifestation	NN	O	O
of	NN	O	O
their	NN	O	O
disease	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

001	NN	O	O
001	NN	O	O
.	NN	O	O

None	NN	O	O
had	NN	O	O
chronic	NN	O	O
or	NN	O	O
prolonged	NN	O	O
manifestations	NN	O	O
of	NN	O	O
FMF	NN	O	B-Disease
,	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
amyloidosis	NN	O	B-Disease
,	NN	O	O
chronic	NN	O	B-Disease
arthritis	NN	O	I-Disease
,	NN	O	O
or	NN	O	O
protracted	NN	O	O
myalgia	NN	O	B-Disease
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

001	NN	O	O
.	NN	O	O

The	NN	O	O
response	NN	O	O
to	NN	O	O
treatment	NN	O	O
was	NN	O	O
good	NN	O	O
despite	NN	O	O
using	NN	O	O
low	NN	O	O
colchicine	NN	O	O
dose	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

05	NN	O	O
.	NN	O	O

The	NN	O	O
overall	NN	O	O
severity	NN	O	O
score	NN	O	O
indicated	NN	O	O
a	NN	O	O
mild	NN	O	O
disease	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

001	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
absence	NN	O	O
of	NN	O	O
M694V	NN	O	O
homozygosity	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O

01	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
our	NN	O	O
regular	NN	O	O
FMF	NN	O	B-Disease
population	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
FMF	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
late	NN	O	O
age	NN	O	O
defines	NN	O	O
a	NN	O	O
milder	NN	O	O
form	NN	O	O
of	NN	O	O
disease	NN	O	O
with	NN	O	O
typical	NN	O	O
clinical	NN	O	O
,	NN	O	O
demographic	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
characteristics	NN	O	O

A	NN	O	O
highly	NN	O	O
accurate	NN	O	O
,	NN	O	O
low	NN	O	O
cost	NN	O	O
test	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
widespread	NN	O	O
use	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
testing	NN	O	O
has	NN	O	O
been	NN	O	O
limited	NN	O	O
to	NN	O	O
date	NN	O	O
by	NN	O	O
three	NN	O	O
principal	NN	O	O
concerns	NN	O	O
the	NN	O	O
fear	NN	O	O
of	NN	O	O
loss	NN	O	O
of	NN	O	O
health	NN	O	O
and	NN	O	O
life	NN	O	O
insurance	NN	O	O
,	NN	O	O
the	NN	O	O
uncertain	NN	O	O
clinical	NN	O	O
value	NN	O	O
of	NN	O	O
a	NN	O	O
positive	NN	O	O
test	NN	O	O
result	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
current	NN	O	O
lack	NN	O	O
of	NN	O	O
an	NN	O	O
inexpensive	NN	O	O
and	NN	O	O
sensitive	NN	O	O
screening	NN	O	O
test	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
an	NN	O	O
inexpensive	NN	O	O
system	NN	O	O
for	NN	O	O
gene	NN	O	O
mutational	NN	O	O
scanning	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
extensive	NN	O	O
multiplex	NN	O	O
PCR	NN	O	O
amplification	NN	O	O
and	NN	O	O
two	NN	O	O
dimensional	NN	O	O
electrophoresis	NN	O	O
.	NN	O	O

The	NN	O	O
efficiency	NN	O	O
of	NN	O	O
this	NN	O	O
system	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
screening	NN	O	O
test	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
,	NN	O	O
was	NN	O	O
evaluated	NN	O	O
in	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
60	NN	O	O
samples	NN	O	O
from	NN	O	O
high	NN	O	O
risk	NN	O	O
women	NN	O	O
,	NN	O	O
14	NN	O	O
of	NN	O	O
which	NN	O	O
contained	NN	O	O
a	NN	O	O
previously	NN	O	O
identified	NN	O	O
mutation	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

All	NN	O	O
14	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
additional	NN	O	O
five	NN	O	O
that	NN	O	O
had	NN	O	O
previously	NN	O	O
escaped	NN	O	O
detection	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
19	NN	O	O
mutations	NN	O	O
,	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
15	NN	O	O
different	NN	O	O
polymorphic	NN	O	O
variants	NN	O	O
were	NN	O	O
scored	NN	O	O
,	NN	O	O
most	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
recurring	NN	O	O
.	NN	O	O

All	NN	O	O
were	NN	O	O
confirmed	NN	O	O
by	NN	O	O
nucleotide	NN	O	O
sequencing	NN	O	O
.	NN	O	O

The	NN	O	O
cost	NN	O	O
of	NN	O	O
screening	NN	O	O
per	NN	O	O
sample	NN	O	O
was	NN	O	O
calculated	NN	O	O
to	NN	O	O
be	NN	O	O
approximately	NN	O	O
US	NN	O	O
$	NN	O	O
70	NN	O	O
for	NN	O	O
the	NN	O	O
manual	NN	O	O
technique	NN	O	O
used	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
reduced	NN	O	O
to	NN	O	O
approximately	NN	O	O
US	NN	O	O
$	NN	O	O
10	NN	O	O
with	NN	O	O
the	NN	O	O
introduction	NN	O	O
of	NN	O	O
commercially	NN	O	O
available	NN	O	O
PCR	NN	O	O
robotics	NN	O	O
and	NN	O	O
fluorescent	NN	O	O
imaging	NN	O	O
.	NN	O	O

Implementation	NN	O	O
of	NN	O	O
this	NN	O	O
method	NN	O	O
of	NN	O	O
mutation	NN	O	O
screening	NN	O	O
in	NN	O	O
the	NN	O	O
research	NN	O	O
and	NN	O	O
clinical	NN	O	O
setting	NN	O	O
should	NN	O	O
permit	NN	O	O
rapid	NN	O	O
accrual	NN	O	O
of	NN	O	O
quantitative	NN	O	O
data	NN	O	O
on	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
associations	NN	O	O
for	NN	O	O
the	NN	O	O
evaluation	NN	O	O
of	NN	O	O
diagnostic	NN	O	O
testing	NN	O	O
.	NN	O	O
.	NN	O	O

Maternal	NN	O	B-Disease
uniparental	NN	O	I-Disease
disomy	NN	O	I-Disease
for	NN	O	I-Disease
chromosome	NN	O	I-Disease
14	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
a	NN	O	O
normal	NN	O	O
karyotype	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
a	NN	O	O
maternal	NN	O	B-Disease
uniparental	NN	O	I-Disease
disomy	NN	O	I-Disease
for	NN	O	I-Disease
chromosome	NN	O	I-Disease
14	NN	O	I-Disease
(	NN	O	O
UPD	NN	O	B-Disease
(	NN	O	O
14	NN	O	O
)	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
7	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
he	NN	O	O
was	NN	O	O
referred	NN	O	O
to	NN	O	O
us	NN	O	O
by	NN	O	O
the	NN	O	O
paediatrician	NN	O	O
because	NN	O	O
of	NN	O	O
symptoms	NN	O	O
of	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

He	NN	O	O
showed	NN	O	O
short	NN	O	B-Disease
stature	NN	O	I-Disease
,	NN	O	O
obesity	NN	O	B-Disease
,	NN	O	O
mild	NN	O	O
developmental	NN	O	B-Disease
delay	NN	O	I-Disease
,	NN	O	O
cryptorchidism	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
some	NN	O	O
mild	NN	O	O
dysmorphic	NN	O	B-Disease
features	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
history	NN	O	O
further	NN	O	O
indicated	NN	O	O
intrauterine	NN	O	B-Disease
growth	NN	O	I-Disease
retardation	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

His	NN	O	O
mother	NN	O	O
was	NN	O	O
44	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
his	NN	O	O
birth	NN	O	O
.	NN	O	O

After	NN	O	O
birth	NN	O	O
he	NN	O	O
showed	NN	O	O
hypotonia	NN	O	B-Disease
with	NN	O	O
poor	NN	O	O
sucking	NN	O	O
,	NN	O	O
for	NN	O	O
which	NN	O	O
gavage	NN	O	O
feeding	NN	O	O
was	NN	O	O
needed	NN	O	O
.	NN	O	O

Motor	NN	O	O
development	NN	O	O
was	NN	O	O
delayed	NN	O	O
.	NN	O	O

After	NN	O	O
1	NN	O	O
year	NN	O	O
he	NN	O	O
became	NN	O	O
obese	NN	O	B-Disease
despite	NN	O	O
a	NN	O	O
normal	NN	O	O
appetite	NN	O	O
.	NN	O	O

Recurrent	NN	O	O
middle	NN	O	B-Disease
ear	NN	O	I-Disease
infections	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
high	NN	O	O
pain	NN	O	O
threshold	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
great	NN	O	O
skill	NN	O	O
with	NN	O	O
jigsaw	NN	O	O
puzzles	NN	O	O
were	NN	O	O
reported	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
behavioural	NN	O	O
problems	NN	O	O
or	NN	O	O
sleep	NN	O	O
disturbance	NN	O	O
.	NN	O	O

Chromosomal	NN	O	O
analysis	NN	O	O
was	NN	O	O
normal	NN	O	O
(	NN	O	O
46	NN	O	O
,	NN	O	O
XY	NN	O	O
)	NN	O	O
.	NN	O	O

DNA	NN	O	O
analysis	NN	O	O
for	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
showed	NN	O	O
no	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

Two	NN	O	O
years	NN	O	O
later	NN	O	O
he	NN	O	O
was	NN	O	O
re	NN	O	O
-	NN	O	O
examined	NN	O	O
because	NN	O	O
we	NN	O	O
thought	NN	O	O
his	NN	O	O
features	NN	O	O
fitted	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
-	NN	O	O
like	NN	O	O
phenotype	NN	O	O
associated	NN	O	O
with	NN	O	O
maternal	NN	O	O
UPD	NN	O	B-Disease
(	NN	O	O
14	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
that	NN	O	O
time	NN	O	O
precocious	NN	O	O
puberty	NN	O	O
was	NN	O	O
evident	NN	O	O
.	NN	O	O

DNA	NN	O	O
analysis	NN	O	O
showed	NN	O	O
maternal	NN	O	B-Disease
heterodisomy	NN	O	I-Disease
for	NN	O	O
chromosome	NN	O	O
14	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
11	NN	O	O
cases	NN	O	O
with	NN	O	O
maternal	NN	O	O
UPD	NN	O	B-Disease
(	NN	O	O
14	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
Robertsonian	NN	O	O
translocation	NN	O	O
involving	NN	O	O
chromosome	NN	O	O
14	NN	O	O
was	NN	O	O
detected	NN	O	O
cytogenetically	NN	O	O
before	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
maternal	NN	O	O
UPD	NN	O	B-Disease
(	NN	O	O
14	NN	O	O
)	NN	O	O
based	NN	O	O
on	NN	O	O
clinical	NN	O	O
features	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
underlines	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
for	NN	O	O
maternal	NN	O	O
UPD	NN	O	O
(	NN	O	O
14	NN	O	O
)	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
similar	NN	O	O
PWS	NN	O	B-Disease
-	NN	O	O
like	NN	O	O
phenotype	NN	O	O
even	NN	O	O
without	NN	O	O
previous	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
Robertsonian	NN	O	O
translocation	NN	O	O
involving	NN	O	O
chromosome	NN	O	O
14	NN	O	O
.	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
genetic	NN	O	O
study	NN	O	O
of	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
in	NN	O	O
an	NN	O	O
Australian	NN	O	O
population	NN	O	O
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
FRDA	NN	O	B-Disease
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
210	NN	O	O
-	NN	O	O
amino	NN	O	O
acid	NN	O	O
protein	NN	O	O
called	NN	O	O
frataxin	NN	O	O
.	NN	O	O

An	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
GAA	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
intron	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
more	NN	O	O
than	NN	O	O
95	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
83	NN	O	O
people	NN	O	O
we	NN	O	O
studied	NN	O	O
who	NN	O	O
have	NN	O	O
mutations	NN	O	O
in	NN	O	O
FRDA	NN	O	O
,	NN	O	O
78	NN	O	O
are	NN	O	O
homozygous	NN	O	O
for	NN	O	O
an	NN	O	O
expanded	NN	O	O
GAA	NN	O	O
repeat	NN	O	O
;	NN	O	O
the	NN	O	O
other	NN	O	O
five	NN	O	O
patients	NN	O	O
have	NN	O	O
an	NN	O	O
expansion	NN	O	O
in	NN	O	O
one	NN	O	O
allele	NN	O	O
and	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
detailed	NN	O	O
clinical	NN	O	O
and	NN	O	O
genetic	NN	O	O
study	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
51	NN	O	O
patients	NN	O	O
homozygous	NN	O	O
for	NN	O	O
an	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
GAA	NN	O	O
repeat	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
smaller	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
expanded	NN	O	O
alleles	NN	O	O
and	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
,	NN	O	O
age	NN	O	O
into	NN	O	O
wheelchair	NN	O	O
,	NN	O	O
scoliosis	NN	O	B-Disease
,	NN	O	O
impaired	NN	O	O
vibration	NN	O	O
sense	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
foot	NN	O	B-Disease
deformity	NN	O	I-Disease
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
smaller	NN	O	O
allele	NN	O	O
and	NN	O	O
cardiomyopathy	NN	O	B-Disease
,	NN	O	O
diabetes	NN	O	B-Disease
mellitus	NN	O	I-Disease
,	NN	O	O
loss	NN	O	B-Disease
of	NN	O	I-Disease
proprioception	NN	O	I-Disease
,	NN	O	O
or	NN	O	O
bladder	NN	O	B-Disease
symptoms	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
larger	NN	O	O
allele	NN	O	O
size	NN	O	O
correlated	NN	O	O
with	NN	O	O
bladder	NN	O	B-Disease
symptoms	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
foot	NN	O	B-Disease
deformity	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
duration	NN	O	O
of	NN	O	O
disease	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
wheelchair	NN	O	O
use	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
diabetes	NN	O	B-Disease
,	NN	O	O
scoliosis	NN	O	B-Disease
,	NN	O	O
bladder	NN	O	B-Disease
symptoms	NN	O	I-Disease
and	NN	O	O
impaired	NN	O	B-Disease
proprioception	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
vibration	NN	O	O
sense	NN	O	O
but	NN	O	O
no	NN	O	O
other	NN	O	O
complications	NN	O	O
studied	NN	O	O
.	NN	O	O
.	NN	O	O

Novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
XLRS1	NN	O	O
causing	NN	O	O
retinoschisis	NN	O	B-Disease
,	NN	O	O
including	NN	O	O
first	NN	O	O
evidence	NN	O	O
of	NN	O	O
putative	NN	O	O
leader	NN	O	O
sequence	NN	O	O
change	NN	O	O
.	NN	O	O

Juvenile	NN	O	B-Disease
retinoschisis	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
screened	NN	O	O
31	NN	O	O
new	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
and	NN	O	O
families	NN	O	O
for	NN	O	O
XLRS1	NN	O	O
mutations	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
previously	NN	O	O
reported	NN	O	O
mutations	NN	O	O
for	NN	O	O
60	NN	O	O
of	NN	O	O
our	NN	O	O
families	NN	O	O
(	NN	O	O
Retinoschisis	NN	O	B-Disease
Consortium	NN	O	O
,	NN	O	O
Hum	NN	O	O
Mol	NN	O	O
Genet	NN	O	O
1998	NN	O	O
;	NN	O	O
7	NN	O	O
1185	NN	O	O
-	NN	O	O
1192	NN	O	O
)	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
three	NN	O	O
different	NN	O	O
mutations	NN	O	O
including	NN	O	O
12	NN	O	O
novel	NN	O	O
ones	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
28	NN	O	O
patients	NN	O	O
.	NN	O	O

Mutations	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
include	NN	O	O
19	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
two	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
one	NN	O	O
intragenic	NN	O	O
deletion	NN	O	O
,	NN	O	O
four	NN	O	O
microdeletions	NN	O	O
,	NN	O	O
one	NN	O	O
insertion	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
intronic	NN	O	O
sequence	NN	O	O
substitution	NN	O	O
that	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
splice	NN	O	O
site	NN	O	O
defect	NN	O	O
.	NN	O	O

Two	NN	O	O
novel	NN	O	O
mutations	NN	O	O
,	NN	O	O
c	NN	O	O
.	NN	O	O

38T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
(	NN	O	O
L13P	NN	O	O
)	NN	O	O
and	NN	O	O
c	NN	O	O
.	NN	O	O

667T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
(	NN	O	O
C223R	NN	O	O
)	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
present	NN	O	O
the	NN	O	O
first	NN	O	O
genetic	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
leader	NN	O	O
peptide	NN	O	O
sequence	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
functional	NN	O	O
significance	NN	O	O
at	NN	O	O
the	NN	O	O
carboxyl	NN	O	O
terminal	NN	O	O
of	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
protein	NN	O	O
beyond	NN	O	O
the	NN	O	O
discoidin	NN	O	O
domain	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
25	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
were	NN	O	O
localized	NN	O	O
to	NN	O	O
exons	NN	O	O
4	NN	O	O
-	NN	O	O
6	NN	O	O
,	NN	O	O
emphasizing	NN	O	O
the	NN	O	O
critical	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
the	NN	O	O
discoidin	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
protein	NN	O	O

Growth	NN	O	O
hormone	NN	O	O
treatment	NN	O	O
increases	NN	O	O
CO	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
response	NN	O	O
,	NN	O	O
ventilation	NN	O	O
and	NN	O	O
central	NN	O	O
inspiratory	NN	O	O
drive	NN	O	O
in	NN	O	O
children	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
studied	NN	O	O
whether	NN	O	O
the	NN	O	O
beneficial	NN	O	O
effects	NN	O	O
of	NN	O	O
growth	NN	O	O
hormone	NN	O	O
(	NN	O	O
GH	NN	O	O
)	NN	O	O
treatment	NN	O	O
on	NN	O	O
growth	NN	O	O
and	NN	O	O
body	NN	O	O
composition	NN	O	O
in	NN	O	O
PWS	NN	O	B-Disease
are	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
improvement	NN	O	O
in	NN	O	O
respiratory	NN	O	O
function	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
resting	NN	O	O
ventilation	NN	O	O
,	NN	O	O
airway	NN	O	O
occlusion	NN	O	O
pressure	NN	O	O
(	NN	O	O
P	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
)	NN	O	O
and	NN	O	O
ventilatory	NN	O	O
response	NN	O	O
to	NN	O	O
CO	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
in	NN	O	O
nine	NN	O	O
children	NN	O	O
,	NN	O	O
aged	NN	O	O
7	NN	O	O
-	NN	O	O
14	NN	O	O
years	NN	O	O
,	NN	O	O
before	NN	O	O
and	NN	O	O
6	NN	O	O
-	NN	O	O
9	NN	O	O
months	NN	O	O
after	NN	O	O
the	NN	O	O
start	NN	O	O
of	NN	O	O
GH	NN	O	O
treatment	NN	O	O
.	NN	O	O

During	NN	O	O
GH	NN	O	O
treatment	NN	O	O
,	NN	O	O
resting	NN	O	O
ventilation	NN	O	O
increased	NN	O	O
by	NN	O	O
26	NN	O	O
%	NN	O	O
,	NN	O	O
P	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
by	NN	O	O
72	NN	O	O
%	NN	O	O
and	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
CO	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
by	NN	O	O
65	NN	O	O
%	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
002	NN	O	O
,	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
04	NN	O	O
and	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
02	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
observed	NN	O	O
increase	NN	O	O
in	NN	O	O
ventilatory	NN	O	O
output	NN	O	O
was	NN	O	O
not	NN	O	O
correlated	NN	O	O
to	NN	O	O
changes	NN	O	O
in	NN	O	O
body	NN	O	O
mass	NN	O	O
index	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
Treatment	NN	O	O
of	NN	O	O
children	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
seems	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
central	NN	O	O
respiratory	NN	O	O
structures	NN	O	O
.	NN	O	O

The	NN	O	O
observed	NN	O	O
increase	NN	O	O
in	NN	O	O
ventilation	NN	O	O
and	NN	O	O
inspiratory	NN	O	O
drive	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
improved	NN	O	O
activity	NN	O	O
level	NN	O	O
reported	NN	O	O
by	NN	O	O
parents	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
children	NN	O	O
during	NN	O	O
growth	NN	O	O
hormone	NN	O	O
therapy	NN	O	O

G130V	NN	O	O
,	NN	O	O
a	NN	O	O
common	NN	O	O
FRDA	NN	O	B-Disease
point	NN	O	O
mutation	NN	O	O
,	NN	O	O
appears	NN	O	O
to	NN	O	O
have	NN	O	O
arisen	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
founder	NN	O	O
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
inherited	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

About	NN	O	O
98	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
have	NN	O	O
an	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
GAA	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
intron	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
gene	NN	O	O
,	NN	O	O
FRDA	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
2	NN	O	O
%	NN	O	O
are	NN	O	O
point	NN	O	O
mutations	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
17	NN	O	O
point	NN	O	O
mutations	NN	O	O
so	NN	O	O
far	NN	O	O
described	NN	O	O
,	NN	O	O
three	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
common	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
is	NN	O	O
the	NN	O	O
G130V	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
4	NN	O	O
of	NN	O	O
FRDA	NN	O	O
.	NN	O	O

G130V	NN	O	O
,	NN	O	O
when	NN	O	O
present	NN	O	O
with	NN	O	O
an	NN	O	O
expanded	NN	O	O
GAA	NN	O	O
repeat	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
allele	NN	O	O
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
atypical	NN	O	O
FRDA	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
analysis	NN	O	O
was	NN	O	O
undertaken	NN	O	O
on	NN	O	O
the	NN	O	O
four	NN	O	O
families	NN	O	O
who	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
with	NN	O	O
this	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
common	NN	O	O
founder	NN	O	O
for	NN	O	O
this	NN	O	O
mutation	NN	O	O
.	NN	O	O

Although	NN	O	O
marked	NN	O	O
differences	NN	O	O
in	NN	O	O
extragenic	NN	O	O
marker	NN	O	O
haplotypes	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
,	NN	O	O
similar	NN	O	O
intragenic	NN	O	O
haplotyping	NN	O	O
suggests	NN	O	O
the	NN	O	O
same	NN	O	O
mutation	NN	O	O
founder	NN	O	O
for	NN	O	O
this	NN	O	O
family	NN	O	O
with	NN	O	O
the	NN	O	O
differences	NN	O	O
explicable	NN	O	O
by	NN	O	O
two	NN	O	O
recombination	NN	O	O
events	NN	O	O
.	NN	O	O
.	NN	O	O

Synergistic	NN	O	O
effect	NN	O	O
of	NN	O	O
histone	NN	O	O
hyperacetylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
demethylation	NN	O	O
in	NN	O	O
the	NN	O	O
reactivation	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
.	NN	O	O

Most	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
patients	NN	O	O
have	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
(	NN	O	O
CGG	NN	O	O
)	NN	O	O
(	NN	O	O
n	NN	O	O
)	NN	O	O
sequence	NN	O	O
with	NN	O	O
>	NN	O	O
200	NN	O	O
repeats	NN	O	O
(	NN	O	O
full	NN	O	O
mutation	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
condition	NN	O	O
.	NN	O	O

Hypermethylation	NN	O	O
of	NN	O	O
the	NN	O	O
expanded	NN	O	O
repeat	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
promoter	NN	O	O
is	NN	O	O
almost	NN	O	O
always	NN	O	O
present	NN	O	O
and	NN	O	O
apparently	NN	O	O
suppresses	NN	O	O
transcription	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
showed	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
reactivation	NN	O	O
of	NN	O	O
FMR1	NN	O	O
full	NN	O	O
mutations	NN	O	O
can	NN	O	O
be	NN	O	O
achieved	NN	O	O
by	NN	O	O
inducing	NN	O	O
DNA	NN	O	O
demethylation	NN	O	O
with	NN	O	O
5	NN	O	O
-	NN	O	O
azadeoxycytidine	NN	O	O
(	NN	O	O
5	NN	O	O
-	NN	O	O
azadC	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
histone	NN	O	O
acetylation	NN	O	O
is	NN	O	O
another	NN	O	O
important	NN	O	O
factor	NN	O	O
in	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
;	NN	O	O
therefore	NN	O	O
,	NN	O	O
we	NN	O	O
treated	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
lines	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
mosaic	NN	O	O
full	NN	O	O
mutation	NN	O	O
patients	NN	O	O
with	NN	O	O
three	NN	O	O
drugs	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
histone	NN	O	O
hyperacetylation	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
a	NN	O	O
consistent	NN	O	O
,	NN	O	O
although	NN	O	O
modest	NN	O	O
,	NN	O	O
reactivation	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
with	NN	O	O
4	NN	O	O
-	NN	O	O
phenylbutyrate	NN	O	O
,	NN	O	O
sodium	NN	O	O
butyrate	NN	O	O
and	NN	O	O
trichostatin	NN	O	O
A	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
combining	NN	O	O
these	NN	O	O
drugs	NN	O	O
with	NN	O	O
5	NN	O	O
-	NN	O	O
azadC	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
2	NN	O	O
-	NN	O	O
to	NN	O	O
5	NN	O	O
-	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
FMR1	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
obtained	NN	O	O
with	NN	O	O
5	NN	O	O
-	NN	O	O
azadC	NN	O	O
alone	NN	O	O
,	NN	O	O
thus	NN	O	O
showing	NN	O	O
a	NN	O	O
marked	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
of	NN	O	O
histone	NN	O	O
hyperacetylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
demethylation	NN	O	O
in	NN	O	O
the	NN	O	O
reactivation	NN	O	O
of	NN	O	O
FMR1	NN	O	O
full	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Constitutional	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	O
VHL	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
deletions	NN	O	O
detected	NN	O	O
in	NN	O	O
VHL	NN	O	B-Disease
families	NN	O	O
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	I-Disease
VHL	NN	O	I-Disease
)	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominantly	NN	O	I-Disease
inherited	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
predisposing	NN	O	O
to	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
tumor	NN	O	B-Disease
types	NN	O	O
that	NN	O	O
include	NN	O	O
retinal	NN	O	O
hemangioblastomas	NN	O	B-Disease
,	NN	O	O
hemangioblastomas	NN	O	B-Disease
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinomas	NN	O	I-Disease
,	NN	O	O
pancreatic	NN	O	B-Disease
cysts	NN	O	I-Disease
and	NN	O	I-Disease
tumors	NN	O	I-Disease
,	NN	O	O
pheochromocytomas	NN	O	B-Disease
,	NN	O	O
endolymphatic	NN	O	B-Disease
sac	NN	O	I-Disease
tumors	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
epididymal	NN	O	B-Disease
cystadenomas	NN	O	I-Disease
[	NN	O	O
W	NN	O	O
.	NN	O	O

M	NN	O	O
.	NN	O	O

Linehan	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O

Am	NN	O	O
.	NN	O	O
Med	NN	O	O
.	NN	O	O
Assoc	NN	O	O
.	NN	O	O
,	NN	O	O
273	NN	O	O
564	NN	O	O
-	NN	O	O
570	NN	O	O
,	NN	O	O
1995	NN	O	O
;	NN	O	O
E	NN	O	O
.	NN	O	O
A	NN	O	O
.	NN	O	O
Maher	NN	O	O
and	NN	O	O
W	NN	O	O
.	NN	O	O
G	NN	O	O
.	NN	O	O
Kaelin	NN	O	O
,	NN	O	O
Jr	NN	O	O
.	NN	O	O
,	NN	O	O
Medicine	NN	O	O
(	NN	O	O
Baltimore	NN	O	O
)	NN	O	O
,	NN	O	O
76	NN	O	O
381	NN	O	O
-	NN	O	O
391	NN	O	O
,	NN	O	O
1997	NN	O	O
;	NN	O	O
W	NN	O	O
.	NN	O	O
M	NN	O	O
.	NN	O	O
Linehan	NN	O	O
and	NN	O	O
R	NN	O	O
.	NN	O	O
D	NN	O	O
.	NN	O	O
Klausner	NN	O	O
,	NN	O	O
In	NN	O	O
B	NN	O	O
.	NN	O	O
Vogelstein	NN	O	O
and	NN	O	O
K	NN	O	O
.	NN	O	O
Kinzler	NN	O	O
(	NN	O	O
eds	NN	O	O
.	NN	O	O
)	NN	O	O
,	NN	O	O
The	NN	O	O
Genetic	NN	O	O
Basis	NN	O	O
of	NN	O	O
Human	NN	O	O
Cancer	NN	O	O
,	NN	O	O
pp	NN	O	O
.	NN	O	O

455	NN	O	O
-	NN	O	O
473	NN	O	O
,	NN	O	O
McGraw	NN	O	O
-	NN	O	O
Hill	NN	O	O
,	NN	O	O
1998	NN	O	O
]	NN	O	O
.	NN	O	O

The	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
chromosome	NN	O	O
3p25	NN	O	O
-	NN	O	O
26	NN	O	O
and	NN	O	O
cloned	NN	O	O
[	NN	O	O
F	NN	O	O
.	NN	O	O
Latif	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
Science	NN	O	O
(	NN	O	O
Washington	NN	O	O
DC	NN	O	O
)	NN	O	O
,	NN	O	O
260	NN	O	O
1317	NN	O	O
-	NN	O	O
1320	NN	O	O
,	NN	O	O
1993	NN	O	O
]	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
VHL	NN	O	B-Disease
kindreds	NN	O	O
.	NN	O	O

The	NN	O	O
reported	NN	O	O
frequency	NN	O	O
of	NN	O	O
detection	NN	O	O
of	NN	O	O
VHL	NN	O	B-Disease
germline	NN	O	O
mutations	NN	O	O
has	NN	O	O
varied	NN	O	O
from	NN	O	O
39	NN	O	O
to	NN	O	O
80	NN	O	O
%	NN	O	O
(	NN	O	O
J	NN	O	O
.	NN	O	O
M	NN	O	O
.	NN	O	O
Whaley	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
Am	NN	O	O
.	NN	O	O
J	NN	O	O
.	NN	O	O
Hum	NN	O	O
.	NN	O	O
Genet	NN	O	O
.	NN	O	O
,	NN	O	O
55	NN	O	O
1092	NN	O	O
-	NN	O	O
1102	NN	O	O
,	NN	O	O
1994	NN	O	O
;	NN	O	O
Clinical	NN	O	O
Research	NN	O	O
Group	NN	O	O
for	NN	O	O
Japan	NN	O	O
,	NN	O	O
Hum	NN	O	O
.	NN	O	O
Mol	NN	O	O
.	NN	O	O
Genet	NN	O	O
.	NN	O	O
,	NN	O	O
4	NN	O	O
2233	NN	O	O
-	NN	O	O
2237	NN	O	O
,	NN	O	O
1995	NN	O	O
;	NN	O	O
F	NN	O	O
.	NN	O	O
Chen	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
Hum	NN	O	O
.	NN	O	O
Mutat	NN	O	O
.	NN	O	O
,	NN	O	O
5	NN	O	O
66	NN	O	O
-	NN	O	O
75	NN	O	O
,	NN	O	O
1995	NN	O	O
;	NN	O	O
E	NN	O	O
.	NN	O	O
R	NN	O	O
.	NN	O	O
Maher	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O
Med	NN	O	O
.	NN	O	O
Genet	NN	O	O
.	NN	O	O
,	NN	O	O
33	NN	O	O
328	NN	O	O
-	NN	O	O
332	NN	O	O
,	NN	O	O
1996	NN	O	O
;	NN	O	O
B	NN	O	O
.	NN	O	O
Zbar	NN	O	O
,	NN	O	O
Cancer	NN	O	O
Surv	NN	O	O
.	NN	O	O
,	NN	O	O
25	NN	O	O
219	NN	O	O
-	NN	O	O
232	NN	O	O
,	NN	O	O
1995	NN	O	O
)	NN	O	O
.	NN	O	O

Recently	NN	O	O
a	NN	O	O
quantitative	NN	O	O
Southern	NN	O	O
blotting	NN	O	O
procedure	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
improve	NN	O	O
this	NN	O	O
frequency	NN	O	O
(	NN	O	O
C	NN	O	O
.	NN	O	O
Stolle	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
Hum	NN	O	O
.	NN	O	O
Mutat	NN	O	O
.	NN	O	O
,	NN	O	O
12	NN	O	O
417	NN	O	O
-	NN	O	O
423	NN	O	O
,	NN	O	O
1998	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
as	NN	O	O
a	NN	O	O
method	NN	O	O
to	NN	O	O
detect	NN	O	O
and	NN	O	O
characterize	NN	O	O
VHL	NN	O	B-Disease
germline	NN	O	O
deletions	NN	O	O
.	NN	O	O

We	NN	O	O
reexamined	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
VHL	NN	O	B-Disease
patients	NN	O	O
shown	NN	O	O
previously	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
and	NN	O	O
sequencing	NN	O	O
analysis	NN	O	O
not	NN	O	O
to	NN	O	O
harbor	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
constitutional	NN	O	O
deletions	NN	O	O
in	NN	O	O
29	NN	O	O
of	NN	O	O
30	NN	O	O
VHL	NN	O	B-Disease
patients	NN	O	O
in	NN	O	O
this	NN	O	O
group	NN	O	O
using	NN	O	O
cosmid	NN	O	O
and	NN	O	O
P1	NN	O	O
probes	NN	O	O
that	NN	O	O
cover	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
tested	NN	O	O
six	NN	O	O
phenotypically	NN	O	O
normal	NN	O	O
offspring	NN	O	O
from	NN	O	O
four	NN	O	O
of	NN	O	O
these	NN	O	O
VHL	NN	O	B-Disease
families	NN	O	O
two	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
the	NN	O	O
deletion	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
four	NN	O	O
were	NN	O	O
deletion	NN	O	O
-	NN	O	O
free	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
germline	NN	O	O
mosaicism	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
.	NN	O	O

In	NN	O	O
sum	NN	O	O
,	NN	O	O
FISH	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
simple	NN	O	O
and	NN	O	O
reliable	NN	O	O
method	NN	O	O
to	NN	O	O
detect	NN	O	O
VHL	NN	O	B-Disease
germline	NN	O	O
deletions	NN	O	O
and	NN	O	O
practically	NN	O	O
useful	NN	O	O
in	NN	O	O
cases	NN	O	O
where	NN	O	O
other	NN	O	O
methods	NN	O	O
of	NN	O	O
screening	NN	O	O
have	NN	O	O
failed	NN	O	O
to	NN	O	O
detect	NN	O	O
a	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	I-Disease
abnormality	NN	O	I-Disease

Spectrum	NN	O	O
of	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
human	NN	O	O
cancer	NN	O	B-Disease
and	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
.	NN	O	O

The	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
gene	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
SWI	NN	O	O
/	NN	O	O
SNF	NN	O	O
chromatin	NN	O	O
ATP	NN	O	O
-	NN	O	O
dependent	NN	O	O
remodeling	NN	O	O
complex	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
new	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
localized	NN	O	O
on	NN	O	O
chromosome	NN	O	O
22q11	NN	O	O
.	NN	O	O

2	NN	O	O
and	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
mutated	NN	O	O
in	NN	O	O
malignant	NN	O	B-Disease
rhabdoid	NN	O	I-Disease
tumors	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
searched	NN	O	O
for	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
mutations	NN	O	O
in	NN	O	O
229	NN	O	O
tumors	NN	O	B-Disease
of	NN	O	O
various	NN	O	O
origins	NN	O	O
using	NN	O	O
a	NN	O	O
screening	NN	O	O
method	NN	O	O
based	NN	O	O
on	NN	O	O
denaturing	NN	O	O
high	NN	O	O
-	NN	O	O
performance	NN	O	O
liquid	NN	O	O
chromatography	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
31	NN	O	O
homozygous	NN	O	O
deletions	NN	O	O
and	NN	O	O
36	NN	O	O
point	NN	O	O
alterations	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
were	NN	O	O
scattered	NN	O	O
along	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
and	NN	O	O
included	NN	O	O
15	NN	O	O
nonsense	NN	O	O
,	NN	O	O
15	NN	O	O
frameshift	NN	O	O
,	NN	O	O
three	NN	O	O
splice	NN	O	O
site	NN	O	O
,	NN	O	O
two	NN	O	O
missense	NN	O	O
and	NN	O	O
one	NN	O	O
editing	NN	O	O
mutations	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
retrieved	NN	O	O
in	NN	O	O
most	NN	O	O
rhabdoid	NN	O	B-Disease
tumors	NN	O	I-Disease
,	NN	O	O
whatever	NN	O	O
their	NN	O	O
sites	NN	O	O
of	NN	O	O
occurrence	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
common	NN	O	O
pathogenetic	NN	O	O
origin	NN	O	O
of	NN	O	O
these	NN	O	O
tumors	NN	O	B-Disease
.	NN	O	O

Recurrent	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
alterations	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
choroid	NN	O	B-Disease
plexus	NN	O	I-Disease
carcinomas	NN	O	I-Disease
and	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
central	NN	O	O
primitive	NN	O	O
neuroectodermal	NN	O	B-Disease
tumors	NN	O	I-Disease
(	NN	O	O
cPNETs	NN	O	O
)	NN	O	O
and	NN	O	O
medulloblastomas	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
point	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
breast	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
Wilms	NN	O	B-Disease
tumors	NN	O	I-Disease
,	NN	O	O
gliomas	NN	O	B-Disease
,	NN	O	O
ependymomas	NN	O	B-Disease
,	NN	O	O
sarcomas	NN	O	B-Disease
and	NN	O	O
other	NN	O	O
tumor	NN	O	B-Disease
types	NN	O	O
,	NN	O	O
even	NN	O	O
though	NN	O	O
most	NN	O	O
analyzed	NN	O	O
cases	NN	O	O
harbored	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
at	NN	O	O
22q11	NN	O	O
.	NN	O	O

2	NN	O	O
loci	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
rhabdoid	NN	O	B-Disease
tumors	NN	O	I-Disease
,	NN	O	O
choroid	NN	O	B-Disease
plexus	NN	O	I-Disease
carcinomas	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
medulloblastomas	NN	O	B-Disease
and	NN	O	O
cPNETs	NN	O	O
share	NN	O	O
common	NN	O	O
pathways	NN	O	O
of	NN	O	O
oncogenesis	NN	O	O
related	NN	O	O
to	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
alteration	NN	O	O
and	NN	O	O
that	NN	O	O
hSNF5	NN	O	O
/	NN	O	O
INI1	NN	O	O
mutations	NN	O	O
define	NN	O	O
a	NN	O	O
genetically	NN	O	O
homogeneous	NN	O	O
family	NN	O	O
of	NN	O	O
highly	NN	O	O
aggressive	NN	O	O
cancers	NN	O	B-Disease
mainly	NN	O	O
occurring	NN	O	O
in	NN	O	O
young	NN	O	O
children	NN	O	O
and	NN	O	O
frequently	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
always	NN	O	O
,	NN	O	O
exhibiting	NN	O	O
a	NN	O	O
rhabdoid	NN	O	B-Disease
phenotype	NN	O	O

Ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
with	NN	O	O
an	NN	O	O
altered	NN	O	O
conformation	NN	O	O
that	NN	O	O
exposes	NN	O	O
the	NN	O	O
polyglutamine	NN	O	O
domain	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
.	NN	O	O

Spinocerebellar	NN	O	B-Disease
ataxia	NN	O	I-Disease
type	NN	O	I-Disease
-	NN	O	I-Disease
3	NN	O	I-Disease
or	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
SCA3	NN	O	B-Disease
/	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
CAG	NN	O	B-Disease
/	NN	O	I-Disease
polyglutamine	NN	O	I-Disease
repeat	NN	O	I-Disease
disease	NN	O	I-Disease
family	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
family	NN	O	O
of	NN	O	O
disorders	NN	O	O
,	NN	O	O
a	NN	O	O
normally	NN	O	O
polymorphic	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
becomes	NN	O	O
expanded	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
expanded	NN	O	O
polyglutamine	NN	O	O
domain	NN	O	O
in	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
product	NN	O	O
.	NN	O	O

Experimental	NN	O	O
models	NN	O	O
of	NN	O	O
polyglutamine	NN	O	B-Disease
disease	NN	O	I-Disease
implicate	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
pathogenesis	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
link	NN	O	O
between	NN	O	O
intranuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
expanded	NN	O	O
polyglutamine	NN	O	O
and	NN	O	O
neuronal	NN	O	B-Disease
dysfunction	NN	O	I-Disease
remains	NN	O	O
unclear	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
,	NN	O	O
the	NN	O	O
disease	NN	O	O
protein	NN	O	O
in	NN	O	O
SCA3	NN	O	B-Disease
/	NN	O	O
MJD	NN	O	B-Disease
,	NN	O	O
adopts	NN	O	O
a	NN	O	O
unique	NN	O	O
conformation	NN	O	O
when	NN	O	O
expressed	NN	O	O
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
transfected	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
1C2	NN	O	O
is	NN	O	O
known	NN	O	O
preferentially	NN	O	O
to	NN	O	O
bind	NN	O	O
expanded	NN	O	O
polyglutamine	NN	O	O
,	NN	O	O
but	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
it	NN	O	O
also	NN	O	O
binds	NN	O	O
a	NN	O	O
fragment	NN	O	O
of	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
containing	NN	O	O
a	NN	O	O
normal	NN	O	O
glutamine	NN	O	O
repeat	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
exposes	NN	O	O
the	NN	O	O
glutamine	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
non	NN	O	O
-	NN	O	O
pathological	NN	O	O
protein	NN	O	O
,	NN	O	O
allowing	NN	O	O
it	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
1C2	NN	O	O
.	NN	O	O

Fractionation	NN	O	O
and	NN	O	O
immunochemical	NN	O	O
experiments	NN	O	O
indicate	NN	O	O
that	NN	O	O
this	NN	O	O
novel	NN	O	O
conformation	NN	O	O
of	NN	O	O
intranuclear	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
is	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
proteolysis	NN	O	O
,	NN	O	O
suggesting	NN	O	O
instead	NN	O	O
that	NN	O	O
association	NN	O	O
with	NN	O	O
nuclear	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
alters	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
which	NN	O	O
exposes	NN	O	O
the	NN	O	O
polyglutamine	NN	O	O
domain	NN	O	O
.	NN	O	O

This	NN	O	O
conformationally	NN	O	O
altered	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
is	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
.	NN	O	O

The	NN	O	O
pathological	NN	O	O
form	NN	O	O
of	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
with	NN	O	O
an	NN	O	O
expanded	NN	O	O
polyglutamine	NN	O	O
domain	NN	O	O
also	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
SCA3	NN	O	B-Disease
/	NN	O	O
MJD	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
an	NN	O	O
altered	NN	O	O
conformation	NN	O	O
of	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
that	NN	O	O
exposes	NN	O	O
the	NN	O	O
polyglutamine	NN	O	O
domain	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
MAGEL2	NN	O	O
gene	NN	O	O
and	NN	O	O
its	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
are	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
and	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
region	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
complex	NN	O	O
neurogenetic	NN	O	B-Disease
disorder	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
phenotype	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
contiguous	NN	O	B-Disease
gene	NN	O	I-Disease
syndrome	NN	O	I-Disease
involving	NN	O	O
genes	NN	O	O
which	NN	O	O
are	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
only	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
region	NN	O	O
.	NN	O	O

Four	NN	O	O
mouse	NN	O	O
models	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
have	NN	O	O
been	NN	O	O
reported	NN	O	O
but	NN	O	O
these	NN	O	O
do	NN	O	O
not	NN	O	O
definitively	NN	O	O
allow	NN	O	O
the	NN	O	O
delineation	NN	O	O
of	NN	O	O
the	NN	O	O
critical	NN	O	O
region	NN	O	O
and	NN	O	O
the	NN	O	O
associated	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
aetiology	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
targeted	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
mouse	NN	O	O
homologues	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
candidate	NN	O	O
PWS	NN	O	B-Disease
genes	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
features	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
new	NN	O	O
genes	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
remains	NN	O	O
crucial	NN	O	O
for	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
this	NN	O	O
manuscript	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
MAGEL2	NN	O	O
and	NN	O	O
its	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
Magel2	NN	O	O
.	NN	O	O

These	NN	O	O
are	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
and	NN	O	O
mouse	NN	O	O
7C	NN	O	O
regions	NN	O	O
,	NN	O	O
in	NN	O	O
close	NN	O	O
proximity	NN	O	O
to	NN	O	O
NDN	NN	O	O
/	NN	O	O
Ndn	NN	O	O
.	NN	O	O

By	NN	O	O
northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
detect	NN	O	O
any	NN	O	O
expression	NN	O	O
of	NN	O	O
MAGEL2	NN	O	O
/	NN	O	O
Magel2	NN	O	O
but	NN	O	O
by	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
analysis	NN	O	O
,	NN	O	O
specific	NN	O	O
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
fetal	NN	O	O
and	NN	O	O
adult	NN	O	O
brain	NN	O	O
and	NN	O	O
in	NN	O	O
placenta	NN	O	O
.	NN	O	O

Both	NN	O	O
genes	NN	O	O
are	NN	O	O
intronless	NN	O	O
with	NN	O	O
tandem	NN	O	O
direct	NN	O	O
repeat	NN	O	O
sequences	NN	O	O
contained	NN	O	O
within	NN	O	O
a	NN	O	O
CpG	NN	O	O
island	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
-	NN	O	O
untranscribed	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
transcripts	NN	O	O
encode	NN	O	O
putative	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
MAGE	NN	O	O
proteins	NN	O	O
and	NN	O	O
NDN	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
MAGEL2	NN	O	O
/	NN	O	O
Magel2	NN	O	O
are	NN	O	O
expressed	NN	O	O
only	NN	O	O
from	NN	O	O
the	NN	O	O
paternal	NN	O	O
allele	NN	O	O
in	NN	O	O
brain	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
aetiology	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
its	NN	O	O
mouse	NN	O	O
model	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O
.	NN	O	O

cDNA	NN	O	O
microarrays	NN	O	O
detect	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
myogenic	NN	O	O
transcription	NN	O	O
program	NN	O	O
by	NN	O	O
the	NN	O	O
PAX3	NN	O	O
-	NN	O	O
FKHR	NN	O	O
fusion	NN	O	O
oncogene	NN	O	O
.	NN	O	O

Alveolar	NN	O	B-Disease
rhabdomyosarcoma	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
aggressive	NN	O	O
pediatric	NN	O	B-Disease
cancer	NN	O	I-Disease
of	NN	O	I-Disease
striated	NN	O	I-Disease
muscle	NN	O	I-Disease
characterized	NN	O	O
in	NN	O	O
60	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
by	NN	O	O
a	NN	O	O
t	NN	O	O
(	NN	O	O
2	NN	O	O
;	NN	O	O
13	NN	O	O
)	NN	O	O
(	NN	O	O
q35	NN	O	O
;	NN	O	O
q14	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
fusion	NN	O	O
of	NN	O	O
PAX3	NN	O	O
,	NN	O	O
a	NN	O	O
developmental	NN	O	O
transcription	NN	O	O
factor	NN	O	O
required	NN	O	O
for	NN	O	O
limb	NN	O	O
myogenesis	NN	O	O
,	NN	O	O
with	NN	O	O
FKHR	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
forkhead	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

The	NN	O	O
resultant	NN	O	O
PAX3	NN	O	O
-	NN	O	O
FKHR	NN	O	O
gene	NN	O	O
possesses	NN	O	O
transforming	NN	O	O
properties	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
this	NN	O	O
chimeric	NN	O	O
oncogene	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
are	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
both	NN	O	O
Pax3	NN	O	O
and	NN	O	O
PAX3	NN	O	O
-	NN	O	O
FKHR	NN	O	O
were	NN	O	O
introduced	NN	O	O
into	NN	O	O
NIH	NN	O	O
3T3	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
resultant	NN	O	O
gene	NN	O	O
expression	NN	O	O
changes	NN	O	O
were	NN	O	O
analyzed	NN	O	O
with	NN	O	O
a	NN	O	O
murine	NN	O	O
cDNA	NN	O	O
microarray	NN	O	O
containing	NN	O	O
2	NN	O	O
,	NN	O	O
225	NN	O	O
elements	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
PAX3	NN	O	O
-	NN	O	O
FKHR	NN	O	O
but	NN	O	O
not	NN	O	O
PAX3	NN	O	O
activated	NN	O	O
a	NN	O	O
myogenic	NN	O	O
transcription	NN	O	O
program	NN	O	O
including	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
MyoD	NN	O	O
,	NN	O	O
Myogenin	NN	O	O
,	NN	O	O
Six1	NN	O	O
,	NN	O	O
and	NN	O	O
Slug	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
battery	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
muscle	NN	O	O
function	NN	O	O
.	NN	O	O

Notable	NN	O	O
among	NN	O	O
this	NN	O	O
group	NN	O	O
were	NN	O	O
the	NN	O	O
growth	NN	O	O
factor	NN	O	O
gene	NN	O	O
Igf2	NN	O	O
and	NN	O	O
its	NN	O	O
binding	NN	O	O
protein	NN	O	O
Igfbp5	NN	O	O
.	NN	O	O

Relevance	NN	O	O
of	NN	O	O
this	NN	O	O
model	NN	O	O
was	NN	O	O
suggested	NN	O	O
by	NN	O	O
verification	NN	O	O
that	NN	O	O
three	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
(	NN	O	O
IGFBP5	NN	O	O
,	NN	O	O
HSIX1	NN	O	O
,	NN	O	O
and	NN	O	O
Slug	NN	O	O
)	NN	O	O
were	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
alveolar	NN	O	B-Disease
rhabdomyosarcoma	NN	O	I-Disease
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
utilizes	NN	O	O
cDNA	NN	O	O
microarrays	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
an	NN	O	O
oncogenic	NN	O	O
transcription	NN	O	O
factor	NN	O	O
and	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
profound	NN	O	O
myogenic	NN	O	O
properties	NN	O	O
of	NN	O	O
PAX3	NN	O	O
-	NN	O	O
FKHR	NN	O	O
in	NN	O	O
NIH	NN	O	O
3T3	NN	O	O
cells	NN	O	O
.	NN	O	O
.	NN	O	O

Experimental	NN	O	O
hemochromatosis	NN	O	B-Disease
due	NN	O	O
to	NN	O	O
MHC	NN	O	O
class	NN	O	O
I	NN	O	O
HFE	NN	O	B-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
immune	NN	O	O
status	NN	O	O
and	NN	O	O
iron	NN	O	O
metabolism	NN	O	O
.	NN	O	O

The	NN	O	O
puzzling	NN	O	O
linkage	NN	O	O
between	NN	O	O
genetic	NN	O	O
hemochromatosis	NN	O	B-Disease
and	NN	O	O
histocompatibility	NN	O	O
loci	NN	O	O
became	NN	O	O
even	NN	O	O
more	NN	O	O
so	NN	O	O
when	NN	O	O
the	NN	O	O
gene	NN	O	O
involved	NN	O	O
,	NN	O	O
HFE	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
well	NN	O	O
defined	NN	O	O
,	NN	O	O
mainly	NN	O	O
peptide	NN	O	O
-	NN	O	O
binding	NN	O	O
,	NN	O	O
MHC	NN	O	O
class	NN	O	O
I	NN	O	O
family	NN	O	O
of	NN	O	O
molecules	NN	O	O
,	NN	O	O
HFE	NN	O	O
seems	NN	O	O
to	NN	O	O
perform	NN	O	O
an	NN	O	O
unusual	NN	O	O
yet	NN	O	O
essential	NN	O	O
function	NN	O	O
.	NN	O	O

As	NN	O	O
yet	NN	O	O
,	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
HFE	NN	O	O
function	NN	O	O
in	NN	O	O
iron	NN	O	O
homeostasis	NN	O	O
is	NN	O	O
only	NN	O	O
partial	NN	O	O
;	NN	O	O
an	NN	O	O
even	NN	O	O
more	NN	O	O
open	NN	O	O
question	NN	O	O
is	NN	O	O
its	NN	O	O
possible	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

To	NN	O	O
advance	NN	O	O
on	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
avenues	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
HFE	NN	O	O
alpha1	NN	O	O
and	NN	O	O
alpha2	NN	O	O
putative	NN	O	O
ligand	NN	O	O
binding	NN	O	O
domains	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

HFE	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
animals	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
set	NN	O	O
of	NN	O	O
metabolic	NN	O	O
and	NN	O	O
immune	NN	O	O
parameters	NN	O	O
.	NN	O	O

Faithfully	NN	O	O
mimicking	NN	O	O
human	NN	O	O
hemochromatosis	NN	O	B-Disease
,	NN	O	O
mice	NN	O	O
homozygous	NN	O	O
for	NN	O	O
this	NN	O	O
deletion	NN	O	O
develop	NN	O	O
iron	NN	O	O
overload	NN	O	O
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
higher	NN	O	O
plasma	NN	O	O
iron	NN	O	O
content	NN	O	O
and	NN	O	O
a	NN	O	O
raised	NN	O	O
transferrin	NN	O	O
saturation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
elevated	NN	O	O
hepatic	NN	O	O
iron	NN	O	O
load	NN	O	O
.	NN	O	O

The	NN	O	O
primary	NN	O	O
defect	NN	O	O
could	NN	O	O
,	NN	O	O
indeed	NN	O	O
,	NN	O	O
be	NN	O	O
traced	NN	O	O
to	NN	O	O
an	NN	O	O
augmented	NN	O	O
duodenal	NN	O	O
iron	NN	O	O
absorption	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
measurement	NN	O	O
of	NN	O	O
the	NN	O	O
gut	NN	O	O
mucosal	NN	O	O
iron	NN	O	O
content	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
iron	NN	O	O
regulatory	NN	O	O
proteins	NN	O	O
allows	NN	O	O
a	NN	O	O
more	NN	O	O
informed	NN	O	O
evaluation	NN	O	O
of	NN	O	O
various	NN	O	O
hypotheses	NN	O	O
regarding	NN	O	O
the	NN	O	O
precise	NN	O	O
role	NN	O	O
of	NN	O	O
HFE	NN	O	O
in	NN	O	O
iron	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
an	NN	O	O
extensive	NN	O	O
phenotyping	NN	O	O
of	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
including	NN	O	O
the	NN	O	O
gut	NN	O	O
provides	NN	O	O
no	NN	O	O
compelling	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
obvious	NN	O	O
immune	NN	O	O
-	NN	O	O
linked	NN	O	O
function	NN	O	O
for	NN	O	O
HFE	NN	O	O
.	NN	O	O
.	NN	O	O

Somatic	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
chromosome	NN	O	O
14	NN	O	O
in	NN	O	O
human	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
genetic	NN	O	B-Disease
disorder	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
immune	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
chromosome	NN	O	B-Disease
instability	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
a	NN	O	O
predisposition	NN	O	O
to	NN	O	O
lymphoid	NN	O	B-Disease
malignancy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
chromosomally	NN	O	O
anomalous	NN	O	O
clones	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
in	NN	O	O
eight	NN	O	O
patients	NN	O	O
with	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
banding	NN	O	O
disclosed	NN	O	O
that	NN	O	O
the	NN	O	O
clones	NN	O	O
are	NN	O	O
consistently	NN	O	O
marked	NN	O	O
by	NN	O	O
structural	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
(	NN	O	O
q	NN	O	O
)	NN	O	O
of	NN	O	O
chromosome	NN	O	O
14	NN	O	O
.	NN	O	O

A	NN	O	O
translocation	NN	O	O
involving	NN	O	O
14q	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
clones	NN	O	O
obtained	NN	O	O
from	NN	O	O
seven	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
patients	NN	O	O
whereas	NN	O	O
a	NN	O	O
ring	NN	O	O
14	NN	O	O
chromosome	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
clone	NN	O	O
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
data	NN	O	O
obtained	NN	O	O
by	NN	O	O
others	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
suggest	NN	O	O
that	NN	O	O
structural	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
14q	NN	O	O
is	NN	O	O
the	NN	O	O
initial	NN	O	O
chromosomal	NN	O	O
change	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
clones	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O

Chromosomes	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
studied	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
chronic	NN	O	B-Disease
lymphocytic	NN	O	I-Disease
leukemia	NN	O	I-Disease
.	NN	O	O

Before	NN	O	O
leukemia	NN	O	B-Disease
was	NN	O	O
diagnosed	NN	O	O
,	NN	O	O
the	NN	O	O
patient	NN	O	O
had	NN	O	O
a	NN	O	O
lymphocyte	NN	O	O
clone	NN	O	O
with	NN	O	O
a	NN	O	O
14q	NN	O	O
translocation	NN	O	O
.	NN	O	O

This	NN	O	O
clone	NN	O	O
appears	NN	O	O
to	NN	O	O
have	NN	O	O
given	NN	O	O
rise	NN	O	O
to	NN	O	O
the	NN	O	O
leukemic	NN	O	B-Disease
cells	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
structural	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
14q	NN	O	O
is	NN	O	O
directly	NN	O	O
related	NN	O	O
to	NN	O	O
abnormal	NN	O	O
growth	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
step	NN	O	O
toward	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-Disease
malignancies	NN	O	I-Disease
.	NN	O	O

Increasing	NN	O	O
evidence	NN	O	O
,	NN	O	O
provided	NN	O	O
by	NN	O	O
others	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
nonrandom	NN	O	O
involvement	NN	O	O
of	NN	O	O
14q	NN	O	O
in	NN	O	O
African	NN	O	O
-	NN	O	O
type	NN	O	O
Burkitts	NN	O	B-Disease
lymphoma	NN	O	I-Disease
and	NN	O	O
other	NN	O	O
lymphoid	NN	O	B-Disease
neoplasms	NN	O	I-Disease
further	NN	O	O
strengthens	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
.	NN	O	O
.	NN	O	O

Exon	NN	O	O
9	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
WT1	NN	O	O
gene	NN	O	O
,	NN	O	O
without	NN	O	O
influencing	NN	O	O
KTS	NN	O	O
splice	NN	O	O
isoforms	NN	O	O
,	NN	O	O
are	NN	O	O
also	NN	O	O
responsible	NN	O	O
for	NN	O	O
Frasier	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
new	NN	O	O
mutations	NN	O	O
in	NN	O	O
exon	NN	O	O
9	NN	O	O
of	NN	O	O
the	NN	O	O
WT1	NN	O	O
gene	NN	O	O
that	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
KTS	NN	O	O
splice	NN	O	O
isoforms	NN	O	O
in	NN	O	O
two	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
with	NN	O	O
Frasier	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
FS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
of	NN	O	O
intron	NN	O	O
9	NN	O	O
inducing	NN	O	O
defective	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
was	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
syndrome	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
found	NN	O	O
in	NN	O	O
our	NN	O	O
cases	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
WT1	NN	O	O
gene	NN	O	O
as	NN	O	O
detected	NN	O	O
in	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
DDS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
previously	NN	O	O
proposed	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
syndromes	NN	O	O
originate	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
WT1	NN	O	B-Disease
gene	NN	O	I-Disease
abnormality	NN	O	I-Disease
.	NN	O	O

From	NN	O	O
a	NN	O	O
molecular	NN	O	O
biological	NN	O	O
point	NN	O	O
of	NN	O	O
view	NN	O	O
,	NN	O	O
we	NN	O	O
concluded	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
diseases	NN	O	O
were	NN	O	O
not	NN	O	O
separable	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
FS	NN	O	B-Disease
should	NN	O	O
be	NN	O	O
included	NN	O	O
as	NN	O	O
an	NN	O	O
atypical	NN	O	O
form	NN	O	O
of	NN	O	O
DDS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
intrafamilial	NN	O	O
variability	NN	O	O
for	NN	O	O
deafness	NN	O	B-Disease
in	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
recessive	NN	O	B-Disease
inherited	NN	O	I-Disease
disorder	NN	O	I-Disease
that	NN	O	O
consists	NN	O	O
of	NN	O	O
developmental	NN	O	B-Disease
abnormalities	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
cochlea	NN	O	I-Disease
,	NN	O	O
sensorineural	NN	O	B-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
diffuse	NN	O	B-Disease
thyroid	NN	O	I-Disease
enlargement	NN	O	I-Disease
(	NN	O	O
goiter	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

This	NN	O	O
disorder	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
of	NN	O	O
hereditary	NN	O	B-Disease
deafness	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
disease	NN	O	O
gene	NN	O	O
(	NN	O	O
PDS	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
7q22	NN	O	O
-	NN	O	O
q31	NN	O	O
,	NN	O	O
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
chloride	NN	O	O
-	NN	O	O
iodide	NN	O	O
transport	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
of	NN	O	O
individual	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
one	NN	O	O
Spanish	NN	O	O
family	NN	O	O
that	NN	O	O
shows	NN	O	O
intrafamilial	NN	O	O
variability	NN	O	O
of	NN	O	O
the	NN	O	O
deafness	NN	O	B-Disease
phenotype	NN	O	O
(	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
profound	NN	O	O
and	NN	O	O
one	NN	O	O
with	NN	O	O
moderate	NN	O	O
-	NN	O	O
severe	NN	O	O
deafness	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
new	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
affecting	NN	O	O
intron	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
639	NN	O	O
+	NN	O	O
7	NN	O	O
.	NN	O	O

RNA	NN	O	O
analysis	NN	O	O
from	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
patients	NN	O	O
showed	NN	O	O
that	NN	O	O
mutation	NN	O	O
639	NN	O	O
+	NN	O	O
7A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
generates	NN	O	O
a	NN	O	O
new	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
an	NN	O	O
mRNA	NN	O	O
with	NN	O	O
an	NN	O	O
insertion	NN	O	O
of	NN	O	O
six	NN	O	O
nucleotides	NN	O	O
from	NN	O	O
intron	NN	O	O
4	NN	O	O
of	NN	O	O
PDS	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
newly	NN	O	O
created	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
compete	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
one	NN	O	O
,	NN	O	O
variations	NN	O	O
of	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
aberrant	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
the	NN	O	O
cochlea	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
variability	NN	O	O
in	NN	O	O
the	NN	O	O
deafness	NN	O	B-Disease
presentation	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
Sjogren	NN	O	B-Disease
-	NN	O	I-Disease
Larsson	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
fatty	NN	O	O
aldehyde	NN	O	O
dehydrogenase	NN	O	O
gene	NN	O	O
.	NN	O	O

Sjogren	NN	O	B-Disease
-	NN	O	I-Disease
Larsson	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
SLS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
ichthyosis	NN	O	B-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
spasticity	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
deficient	NN	O	B-Disease
activity	NN	O	I-Disease
of	NN	O	I-Disease
fatty	NN	O	I-Disease
aldehyde	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	O
FALDH	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
molecular	NN	O	O
defects	NN	O	O
causing	NN	O	O
SLS	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
performed	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
FALDH	NN	O	O
gene	NN	O	O
in	NN	O	O
probands	NN	O	O
from	NN	O	O
63	NN	O	O
kindreds	NN	O	O
with	NN	O	O
SLS	NN	O	B-Disease
.	NN	O	O

Among	NN	O	O
these	NN	O	O
patients	NN	O	O
,	NN	O	O
49	NN	O	O
different	NN	O	O
mutations	NN	O	O
-	NN	O	O
including	NN	O	O
10	NN	O	O
deletions	NN	O	O
,	NN	O	O
2	NN	O	O
insertions	NN	O	O
,	NN	O	O
22	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
,	NN	O	O
3	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
9	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
defects	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
complex	NN	O	O
mutations	NN	O	O
-	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
SLS	NN	O	B-Disease
were	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
mutations	NN	O	O
.	NN	O	O

Nineteen	NN	O	O
of	NN	O	O
the	NN	O	O
missense	NN	O	O
mutations	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
severe	NN	O	O
reduction	NN	O	O
of	NN	O	O
FALDH	NN	O	O
enzyme	NN	O	O
catalytic	NN	O	O
activity	NN	O	O
when	NN	O	O
expressed	NN	O	O
in	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
,	NN	O	O
but	NN	O	O
one	NN	O	O
mutation	NN	O	O
(	NN	O	O
798G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
[	NN	O	O
K266N	NN	O	O
]	NN	O	O
)	NN	O	O
seemed	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
greater	NN	O	O
effect	NN	O	O
on	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
.	NN	O	O

The	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutations	NN	O	O
led	NN	O	O
to	NN	O	O
exon	NN	O	O
skipping	NN	O	O
or	NN	O	O
utilization	NN	O	O
of	NN	O	O
cryptic	NN	O	O
acceptor	NN	O	O
-	NN	O	O
splice	NN	O	O
sites	NN	O	O
.	NN	O	O

Thirty	NN	O	O
-	NN	O	O
seven	NN	O	O
mutations	NN	O	O
were	NN	O	O
private	NN	O	O
,	NN	O	O
and	NN	O	O
12	NN	O	O
mutations	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
probands	NN	O	O
of	NN	O	O
European	NN	O	O
or	NN	O	O
Middle	NN	O	O
Eastern	NN	O	O
descent	NN	O	O
.	NN	O	O

Four	NN	O	O
single	NN	O	O
-	NN	O	O
nucleotide	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
SNPs	NN	O	O
)	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
FALDH	NN	O	O
gene	NN	O	O
.	NN	O	O

At	NN	O	O
least	NN	O	O
four	NN	O	O
of	NN	O	O
the	NN	O	O
common	NN	O	O
mutations	NN	O	O
(	NN	O	O
551C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
,	NN	O	O
682C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
,	NN	O	O
733G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
and	NN	O	O
798	NN	O	O
+	NN	O	O
1delG	NN	O	O
)	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
multiple	NN	O	O
SNP	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
mutations	NN	O	O
originated	NN	O	O
independently	NN	O	O
on	NN	O	O
more	NN	O	O
than	NN	O	O
one	NN	O	O
occasion	NN	O	O
or	NN	O	O
were	NN	O	O
ancient	NN	O	O
SLS	NN	O	B-Disease
genes	NN	O	O
that	NN	O	O
had	NN	O	O
undergone	NN	O	O
intragenic	NN	O	O
recombination	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
SLS	NN	O	B-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
strikingly	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
FALDH	NN	O	O
gene	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
framework	NN	O	O
for	NN	O	O
understanding	NN	O	O
the	NN	O	O
genetic	NN	O	O
basis	NN	O	O
of	NN	O	O
SLS	NN	O	B-Disease
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
DNA	NN	O	O
-	NN	O	O
based	NN	O	O
diagnostic	NN	O	O
tests	NN	O	O
.	NN	O	O
.	NN	O	O

Loss	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
function	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
result	NN	O	O
in	NN	O	O
periodontal	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
palmoplantar	NN	O	B-Disease
keratosis	NN	O	I-Disease
.	NN	O	O

Papillon	NN	O	B-Disease
-	NN	O	I-Disease
Lefevre	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
or	NN	O	O
keratosis	NN	O	B-Disease
palmoplantaris	NN	O	I-Disease
with	NN	O	O
periodontopathia	NN	O	B-Disease
(	NN	O	O
PLS	NN	O	B-Disease
,	NN	O	O
MIM	NN	O	O
245000	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
that	NN	O	O
is	NN	O	O
mainly	NN	O	O
ascertained	NN	O	O
by	NN	O	O
dentists	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
severe	NN	O	O
periodontitis	NN	O	B-Disease
that	NN	O	O
afflicts	NN	O	O
patients	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
deciduous	NN	O	O
and	NN	O	O
permanent	NN	O	O
dentitions	NN	O	O
are	NN	O	O
affected	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
premature	NN	O	O
tooth	NN	O	B-Disease
loss	NN	O	I-Disease
.	NN	O	O

Palmoplantar	NN	O	B-Disease
keratosis	NN	O	I-Disease
,	NN	O	O
varying	NN	O	O
from	NN	O	O
mild	NN	O	O
psoriasiform	NN	O	O
scaly	NN	O	O
skin	NN	O	O
to	NN	O	O
overt	NN	O	O
hyperkeratosis	NN	O	B-Disease
,	NN	O	O
typically	NN	O	O
develops	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
three	NN	O	O
years	NN	O	O
of	NN	O	O
life	NN	O	O
.	NN	O	O

Keratosis	NN	O	B-Disease
also	NN	O	O
affects	NN	O	O
other	NN	O	O
sites	NN	O	O
such	NN	O	O
as	NN	O	O
elbows	NN	O	O
and	NN	O	O
knees	NN	O	O
.	NN	O	O

Most	NN	O	O
PLS	NN	O	B-Disease
patients	NN	O	O
display	NN	O	O
both	NN	O	O
periodontitis	NN	O	B-Disease
and	NN	O	O
hyperkeratosis	NN	O	B-Disease
.	NN	O	O

Some	NN	O	O
patients	NN	O	O
have	NN	O	O
only	NN	O	O
palmoplantar	NN	O	B-Disease
keratosis	NN	O	I-Disease
or	NN	O	O
periodontitis	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
in	NN	O	O
rare	NN	O	O
individuals	NN	O	O
the	NN	O	O
periodontitis	NN	O	B-Disease
is	NN	O	O
mild	NN	O	O
and	NN	O	O
of	NN	O	O
late	NN	O	O
onset	NN	O	O
.	NN	O	O

The	NN	O	O
PLS	NN	O	O
locus	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
11q14	NN	O	O
-	NN	O	O
q21	NN	O	O
(	NN	O	O
refs	NN	O	O
7	NN	O	O
,	NN	O	O
8	NN	O	O
,	NN	O	O
9	NN	O	O
)	NN	O	O
.	NN	O	O

Using	NN	O	O
homozygosity	NN	O	O
mapping	NN	O	O
in	NN	O	O
eight	NN	O	O
small	NN	O	O
consanguineous	NN	O	O
families	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
narrowed	NN	O	O
the	NN	O	O
candidate	NN	O	O
region	NN	O	O
to	NN	O	O
a	NN	O	O
1	NN	O	O
.	NN	O	O

2	NN	O	O
-	NN	O	O
cM	NN	O	O
interval	NN	O	O
between	NN	O	O
D11S4082	NN	O	O
and	NN	O	O
D11S931	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
(	NN	O	O
CTSC	NN	O	O
)	NN	O	O
encoding	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
protease	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
(	NN	O	O
or	NN	O	O
dipeptidyl	NN	O	O
aminopeptidase	NN	O	O
I	NN	O	O
)	NN	O	O
lies	NN	O	O
within	NN	O	O
this	NN	O	O
interval	NN	O	O
.	NN	O	O

We	NN	O	O
defined	NN	O	O
the	NN	O	O
genomic	NN	O	O
structure	NN	O	O
of	NN	O	O
CTSC	NN	O	O
and	NN	O	O
found	NN	O	O
mutations	NN	O	O
in	NN	O	O
all	NN	O	O
eight	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
families	NN	O	O
we	NN	O	O
used	NN	O	O
a	NN	O	O
functional	NN	O	O
assay	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
an	NN	O	O
almost	NN	O	O
total	NN	O	O
loss	NN	O	O
of	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
activity	NN	O	O
in	NN	O	O
PLS	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
reduced	NN	O	O
activity	NN	O	O
in	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
.	NN	O	O

Confirmation	NN	O	O
of	NN	O	O
linkage	NN	O	O
of	NN	O	O
Van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
to	NN	O	O
chromosome	NN	O	O
1q32	NN	O	O
:	NN	O	O
evidence	NN	O	O
of	NN	O	O
association	NN	O	O
with	NN	O	O
STR	NN	O	O
alleles	NN	O	O
suggests	NN	O	O
possible	NN	O	O
unique	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
mutation	NN	O	O
.	NN	O	O

Van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
VWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
craniofacial	NN	O	I-Disease
disorder	NN	O	I-Disease
with	NN	O	O
high	NN	O	O
penetrance	NN	O	O
and	NN	O	O
variable	NN	O	O
expression	NN	O	O
.	NN	O	O

Its	NN	O	O
clinical	NN	O	O
features	NN	O	O
are	NN	O	O
variably	NN	O	O
expressed	NN	O	O
,	NN	O	O
but	NN	O	O
include	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
and	NN	O	O
/	NN	O	O
or	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
,	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
and	NN	O	O
hypodontia	NN	O	B-Disease
.	NN	O	O

All	NN	O	O
VWS	NN	O	B-Disease
families	NN	O	O
studied	NN	O	O
to	NN	O	O
date	NN	O	O
map	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
to	NN	O	O
a	NN	O	O
<	NN	O	O
2	NN	O	O
cM	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
1q32	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
locus	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
is	NN	O	O
to	NN	O	O
refine	NN	O	O
the	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
to	NN	O	O
further	NN	O	O
assess	NN	O	O
possible	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
four	NN	O	O
multiplex	NN	O	O
VWS	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

All	NN	O	O
available	NN	O	O
members	NN	O	O
were	NN	O	O
clinically	NN	O	O
assessed	NN	O	O
and	NN	O	O
genotyped	NN	O	O
for	NN	O	O
19	NN	O	O
short	NN	O	O
tandem	NN	O	O
repeat	NN	O	O
markers	NN	O	O
on	NN	O	O
chromosome	NN	O	O
1	NN	O	O
in	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
candidate	NN	O	O
gene	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
and	NN	O	O
multipoint	NN	O	O
limit	NN	O	O
of	NN	O	O
detection	NN	O	O
(	NN	O	O
LOD	NN	O	O
)	NN	O	O
score	NN	O	O
analyses	NN	O	O
using	NN	O	O
a	NN	O	O
high	NN	O	O
penetrance	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
model	NN	O	O
.	NN	O	O

All	NN	O	O
families	NN	O	O
showed	NN	O	O
positive	NN	O	O
LOD	NN	O	O
scores	NN	O	O
without	NN	O	O
any	NN	O	O
recombination	NN	O	O
in	NN	O	O
the	NN	O	O
candidate	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
largest	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
LOD	NN	O	O
score	NN	O	O
was	NN	O	O
5	NN	O	O
.	NN	O	O

87	NN	O	O
87	NN	O	O
.	NN	O	O

Our	NN	O	O
assay	NN	O	O
method	NN	O	O
for	NN	O	O
short	NN	O	O
tandem	NN	O	O
repeat	NN	O	O
(	NN	O	O
STR	NN	O	O
)	NN	O	O
markers	NN	O	O
provided	NN	O	O
highly	NN	O	O
accurate	NN	O	O
size	NN	O	O
estimation	NN	O	O
of	NN	O	O
marker	NN	O	O
allele	NN	O	O
fragment	NN	O	O
sizes	NN	O	O
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
specific	NN	O	O
alleles	NN	O	O
segregating	NN	O	O
with	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
each	NN	O	O
of	NN	O	O
our	NN	O	O
four	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
a	NN	O	O
striking	NN	O	O
pattern	NN	O	O
of	NN	O	O
STR	NN	O	O
allele	NN	O	O
sharing	NN	O	O
at	NN	O	O
several	NN	O	O
closely	NN	O	O
linked	NN	O	O
loci	NN	O	O
among	NN	O	O
our	NN	O	O
four	NN	O	O
Caucasian	NN	O	O
VWS	NN	O	B-Disease
families	NN	O	O
recruited	NN	O	O
at	NN	O	O
three	NN	O	O
different	NN	O	O
locations	NN	O	O
in	NN	O	O
the	NN	O	O
US	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	O
origin	NN	O	O
for	NN	O	O
a	NN	O	O
mutation	NN	O	O
responsible	NN	O	O
for	NN	O	O
many	NN	O	O
or	NN	O	O
most	NN	O	O
cases	NN	O	O
of	NN	O	O
VWS	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
point	NN	O	O
mutation	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
Met	NN	O	O
on	NN	O	O
the	NN	O	O
alpha	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
integrin	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
new	NN	O	O
human	NN	O	O
platelet	NN	O	O
alloantigen	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
and	NN	O	O
affects	NN	O	O
collagen	NN	O	O
-	NN	O	O
induced	NN	O	O
aggregation	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
platelet	NN	O	O
-	NN	O	O
specific	NN	O	O
alloantigen	NN	O	O
,	NN	O	O
termed	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
severe	NN	O	O
case	NN	O	O
of	NN	O	O
neonatal	NN	O	B-Disease
alloimmune	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
alloantigen	NN	O	O
is	NN	O	O
of	NN	O	O
low	NN	O	O
frequency	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
400	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
German	NN	O	O
population	NN	O	O
.	NN	O	O

Immunochemical	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
epitopes	NN	O	O
reside	NN	O	O
on	NN	O	O
platelet	NN	O	O
glycoprotein	NN	O	O
(	NN	O	O
GP	NN	O	O
)	NN	O	O
Ia	NN	O	O
.	NN	O	O

Nucleotide	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
GPIa	NN	O	O
cDNA	NN	O	O
derived	NN	O	O
from	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
-	NN	O	O
positive	NN	O	O
platelets	NN	O	O
showed	NN	O	O
C	NN	O	O
(	NN	O	O
2531	NN	O	O
)	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
(	NN	O	O
2531	NN	O	O
)	NN	O	O
point	NN	O	O
mutation	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
Met	NN	O	O
dimorphism	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
22	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
-	NN	O	O
negative	NN	O	O
normal	NN	O	O
individuals	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
ACG	NN	O	O
(	NN	O	O
2532	NN	O	O
)	NN	O	O
(	NN	O	O
90	NN	O	O
.	NN	O	O
9	NN	O	O
%	NN	O	O
)	NN	O	O
or	NN	O	O
ACA	NN	O	O
(	NN	O	O
2532	NN	O	O
)	NN	O	O
(	NN	O	O
9	NN	O	O
.	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
establish	NN	O	O
a	NN	O	O
DNA	NN	O	O
typing	NN	O	O
technique	NN	O	O
,	NN	O	O
we	NN	O	O
elucidated	NN	O	O
the	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
GPIa	NN	O	O
gene	NN	O	O
adjacent	NN	O	O
to	NN	O	O
the	NN	O	O
polymorphic	NN	O	O
bases	NN	O	O
.	NN	O	O

The	NN	O	O
introns	NN	O	O
(	NN	O	O
421	NN	O	O
bp	NN	O	O
and	NN	O	O
1	NN	O	O
.	NN	O	O
2	NN	O	O
kb	NN	O	O
)	NN	O	O
encompass	NN	O	O
a	NN	O	O
142	NN	O	O
-	NN	O	O
bp	NN	O	O
exon	NN	O	O
with	NN	O	O
the	NN	O	O
2	NN	O	O
polymorphic	NN	O	O
bases	NN	O	O
2531	NN	O	O
and	NN	O	O
2532	NN	O	O
.	NN	O	O

Polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
on	NN	O	O
DNA	NN	O	O
derived	NN	O	O
from	NN	O	O
100	NN	O	O
donors	NN	O	O
using	NN	O	O
the	NN	O	O
restriction	NN	O	O
enzyme	NN	O	O
Mae	NN	O	O
III	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
Met	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
form	NN	O	O
of	NN	O	O
GPIa	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
stable	NN	O	O
Chinese	NN	O	O
hamster	NN	O	O
ovary	NN	O	O
transfectants	NN	O	O
expressing	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
recombinant	NN	O	O
forms	NN	O	O
of	NN	O	O
GPIa	NN	O	O
showed	NN	O	O
that	NN	O	O
anti	NN	O	O
-	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
exclusively	NN	O	O
reacted	NN	O	O
with	NN	O	O
the	NN	O	O
Glu	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Met	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
Glu	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
Lys	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
isoforms	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
anti	NN	O	O
-	NN	O	O
Br	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
(	NN	O	O
HPA	NN	O	O
-	NN	O	O
5b	NN	O	O
)	NN	O	O
only	NN	O	O
recognized	NN	O	O
the	NN	O	O
Lys	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
form	NN	O	O
,	NN	O	O
whereas	NN	O	O
anti	NN	O	O
-	NN	O	O
Br	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
(	NN	O	O
HPA	NN	O	O
-	NN	O	O
5a	NN	O	O
)	NN	O	O
reacted	NN	O	O
with	NN	O	O
both	NN	O	O
Glu	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
and	NN	O	O
Glu	NN	O	O
(	NN	O	O
505	NN	O	O
)	NN	O	O
Met	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
isoforms	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
Met	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
alloantigenic	NN	O	O
determinants	NN	O	O
,	NN	O	O
whereas	NN	O	O
amino	NN	O	O
acid	NN	O	O
505	NN	O	O
(	NN	O	O
Lys	NN	O	O
or	NN	O	O
Glu	NN	O	O
)	NN	O	O
specifically	NN	O	O
controls	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Br	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
and	NN	O	O
Br	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
epitopes	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Platelet	NN	O	O
aggregation	NN	O	O
responses	NN	O	O
of	NN	O	O
Sit	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
individuals	NN	O	O
were	NN	O	O
diminished	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
collagen	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
Thr	NN	O	O
(	NN	O	O
799	NN	O	O
)	NN	O	O
Met	NN	O	O
mutation	NN	O	O
affects	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
GPIa	NN	O	O
/	NN	O	O
IIa	NN	O	O
complex	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
Papillon	NN	O	B-Disease
-	NN	O	I-Disease
Lefevre	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Papillon	NN	O	B-Disease
-	NN	O	I-Disease
Lefevre	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PLS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterised	NN	O	O
by	NN	O	O
palmoplantar	NN	O	B-Disease
hyperkeratosis	NN	O	I-Disease
and	NN	O	O
severe	NN	O	O
early	NN	O	O
onset	NN	O	O
periodontitis	NN	O	B-Disease
that	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
premature	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
dentitions	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
gene	NN	O	O
locus	NN	O	O
for	NN	O	O
PLS	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
a	NN	O	O
2	NN	O	O
.	NN	O	O

8	NN	O	O
cM	NN	O	O
interval	NN	O	O
on	NN	O	O
chromosome	NN	O	O
11q14	NN	O	O
.	NN	O	O

Correlation	NN	O	O
of	NN	O	O
physical	NN	O	O
and	NN	O	O
genetic	NN	O	O
maps	NN	O	O
of	NN	O	O
this	NN	O	O
interval	NN	O	O
indicate	NN	O	O
it	NN	O	O
includes	NN	O	O
at	NN	O	O
least	NN	O	O
40	NN	O	O
ESTs	NN	O	O
and	NN	O	O
six	NN	O	O
known	NN	O	O
genes	NN	O	O
including	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
protease	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
(	NN	O	O
CTSC	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
CTSC	NN	O	O
message	NN	O	O
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
immune	NN	O	O
cells	NN	O	O
including	NN	O	O
polymorphonuclear	NN	O	O
leucocytes	NN	O	O
,	NN	O	O
macrophages	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
precursors	NN	O	O
.	NN	O	O

By	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
CTSC	NN	O	O
is	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
epithelial	NN	O	O
regions	NN	O	O
commonly	NN	O	O
affected	NN	O	O
by	NN	O	O
PLS	NN	O	B-Disease
,	NN	O	O
including	NN	O	O
the	NN	O	O
palms	NN	O	O
,	NN	O	O
soles	NN	O	O
,	NN	O	O
knees	NN	O	O
,	NN	O	O
and	NN	O	O
oral	NN	O	O
keratinised	NN	O	O
gingiva	NN	O	O
.	NN	O	O

The	NN	O	O
4	NN	O	O
.	NN	O	O

7	NN	O	O
kb	NN	O	O
CTSC	NN	O	O
gene	NN	O	O
consists	NN	O	O
of	NN	O	O
two	NN	O	O
exons	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
CTSC	NN	O	O
from	NN	O	O
subjects	NN	O	O
affected	NN	O	O
with	NN	O	O
PLS	NN	O	B-Disease
from	NN	O	O
five	NN	O	O
consanguineous	NN	O	O
Turkish	NN	O	O
families	NN	O	O
identified	NN	O	O
four	NN	O	O
different	NN	O	O
mutations	NN	O	O
.	NN	O	O

An	NN	O	O
exon	NN	O	O
1	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
(	NN	O	O
856C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
introduces	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
286	NN	O	O
.	NN	O	O

Three	NN	O	O
exon	NN	O	O
2	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
deletion	NN	O	O
(	NN	O	O
2692delA	NN	O	O
)	NN	O	O
of	NN	O	O
codon	NN	O	O
349	NN	O	O
introducing	NN	O	O
a	NN	O	O
frameshift	NN	O	O
and	NN	O	O
premature	NN	O	O
termination	NN	O	O
codon	NN	O	O
,	NN	O	O
a	NN	O	O
2	NN	O	O
bp	NN	O	O
deletion	NN	O	O
(	NN	O	O
2673	NN	O	O
-	NN	O	O
2674delCT	NN	O	O
)	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
introduction	NN	O	O
of	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
343	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
substitution	NN	O	O
in	NN	O	O
codon	NN	O	O
429	NN	O	O
(	NN	O	O
2931G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
)	NN	O	O
introducing	NN	O	O
a	NN	O	O
premature	NN	O	O
termination	NN	O	O
codon	NN	O	O
.	NN	O	O

All	NN	O	O
PLS	NN	O	B-Disease
patients	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
mutations	NN	O	O
inherited	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
ancestor	NN	O	O
.	NN	O	O

Parents	NN	O	O
and	NN	O	O
sibs	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
mutations	NN	O	O
do	NN	O	O
not	NN	O	O
show	NN	O	O
either	NN	O	O
the	NN	O	O
palmoplantar	NN	O	B-Disease
hyperkeratosis	NN	O	I-Disease
or	NN	O	O
severe	NN	O	O
early	NN	O	O
onset	NN	O	O
periodontitis	NN	O	B-Disease
characteristic	NN	O	O
of	NN	O	O
PLS	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
more	NN	O	O
complete	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
physiology	NN	O	O
of	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
carries	NN	O	O
significant	NN	O	O
implications	NN	O	O
for	NN	O	O
understanding	NN	O	O
normal	NN	O	O
and	NN	O	O
abnormal	NN	O	O
skin	NN	O	O
development	NN	O	O
and	NN	O	O
periodontal	NN	O	B-Disease
disease	NN	O	I-Disease
susceptibility	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
HGO	NN	O	O
gene	NN	O	O
in	NN	O	O
Finnish	NN	O	O
alkaptonuria	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Alkaptonuria	NN	O	B-Disease
(	NN	O	O
AKU	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
prototypic	NN	O	O
inborn	NN	O	B-Disease
error	NN	O	I-Disease
of	NN	O	I-Disease
metabolism	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
loss	NN	O	O
of	NN	O	O
function	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
homogentisate	NN	O	O
-	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
-	NN	O	O
dioxygenase	NN	O	O
gene	NN	O	O
(	NN	O	O
HGO	NN	O	O
)	NN	O	O
.	NN	O	O

So	NN	O	O
far	NN	O	O
17	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
characterised	NN	O	O
in	NN	O	O
AKU	NN	O	B-Disease
patients	NN	O	O
of	NN	O	O
different	NN	O	O
ethnic	NN	O	O
origin	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
three	NN	O	O
novel	NN	O	O
mutations	NN	O	O
(	NN	O	O
R58fs	NN	O	O
,	NN	O	O
R330S	NN	O	O
,	NN	O	O
and	NN	O	O
H371R	NN	O	O
)	NN	O	O
and	NN	O	O
one	NN	O	O
common	NN	O	O
AKU	NN	O	B-Disease
mutation	NN	O	O
(	NN	O	O
M368V	NN	O	O
)	NN	O	O
,	NN	O	O
detected	NN	O	O
by	NN	O	O
mutational	NN	O	O
and	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
HGO	NN	O	O
gene	NN	O	O
in	NN	O	O
five	NN	O	O
Finnish	NN	O	O
AKU	NN	O	B-Disease
pedigrees	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
novel	NN	O	O
AKU	NN	O	B-Disease
mutations	NN	O	O
are	NN	O	O
most	NN	O	O
likely	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
Finnish	NN	O	O
population	NN	O	O
and	NN	O	O
have	NN	O	O
originated	NN	O	O
recently	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
identical	NN	O	O
5	NN	O	O
'	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
acceptor	NN	O	O
mutation	NN	O	O
in	NN	O	O
five	NN	O	O
attenuated	NN	O	B-Disease
APC	NN	O	I-Disease
families	NN	O	O
from	NN	O	O
Newfoundland	NN	O	O
demonstrates	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
.	NN	O	O

Inherited	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
predispose	NN	O	O
carriers	NN	O	O
to	NN	O	O
multiple	NN	O	O
adenomatous	NN	O	B-Disease
polyps	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
colon	NN	O	I-Disease
and	NN	O	I-Disease
rectum	NN	O	I-Disease
and	NN	O	O
to	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
extreme	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
less	NN	O	O
severe	NN	O	O
disease	NN	O	O
known	NN	O	O
as	NN	O	O
attenuated	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
AAPC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Many	NN	O	O
individuals	NN	O	O
with	NN	O	O
AAPC	NN	O	B-Disease
develop	NN	O	O
relatively	NN	O	O
few	NN	O	O
colorectal	NN	O	B-Disease
polyps	NN	O	I-Disease
but	NN	O	O
are	NN	O	O
still	NN	O	O
at	NN	O	O
high	NN	O	O
risk	NN	O	O
for	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
5	NN	O	O
APC	NN	O	B-Disease
germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
five	NN	O	O
separately	NN	O	O
ascertained	NN	O	O
AAPC	NN	O	B-Disease
families	NN	O	O
from	NN	O	O
Newfoundland	NN	O	O
,	NN	O	O
Canada	NN	O	O
.	NN	O	O

This	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
single	NN	O	O
basepair	NN	O	O
change	NN	O	O
(	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
splice	NN	O	O
-	NN	O	O
acceptor	NN	O	O
region	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
intron	NN	O	O
3	NN	O	O
that	NN	O	O
creates	NN	O	O
a	NN	O	O
mutant	NN	O	O
RNA	NN	O	O
without	NN	O	O
exon	NN	O	O
4	NN	O	O
of	NN	O	O
APC	NN	O	O
.	NN	O	O

The	NN	O	O
observation	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
APC	NN	O	B-Disease
mutation	NN	O	O
in	NN	O	O
five	NN	O	O
families	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
geographic	NN	O	O
area	NN	O	O
demonstrates	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
this	NN	O	O
germline	NN	O	O
mutation	NN	O	O
strengthens	NN	O	O
the	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
5	NN	O	O
location	NN	O	O
of	NN	O	O
an	NN	O	O
APC	NN	O	B-Disease
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
mutation	NN	O	O
and	NN	O	O
the	NN	O	O
attenuated	NN	O	B-Disease
polyposis	NN	O	I-Disease
phenotype	NN	O	O
.	NN	O	O
.	NN	O	O

Alstrom	NN	O	B-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
2p13	NN	O	O
.	NN	O	O

Alstrom	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
,	NN	O	O
sensorineural	NN	O	B-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
,	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
obesity	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
insulin	NN	O	I-Disease
-	NN	O	I-Disease
dependent	NN	O	I-Disease
diabetes	NN	O	I-Disease
mellitus	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
Alstrom	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ALMS1	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
previously	NN	O	O
localized	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
2p13	NN	O	O
by	NN	O	O
homozygosity	NN	O	O
mapping	NN	O	O
in	NN	O	O
two	NN	O	O
distinct	NN	O	O
isolated	NN	O	O
populations	NN	O	O
-	NN	O	O
French	NN	O	O
Acadian	NN	O	O
and	NN	O	O
North	NN	O	O
African	NN	O	O
.	NN	O	O

Pair	NN	O	O
-	NN	O	O
wise	NN	O	O
analyses	NN	O	O
resulted	NN	O	O
in	NN	O	O
maximum	NN	O	O
lod	NN	O	O
(	NN	O	O
logarithm	NN	O	O
of	NN	O	O
the	NN	O	O
odds	NN	O	O
ratio	NN	O	O
)	NN	O	O
scores	NN	O	O
of	NN	O	O
3	NN	O	O
.	NN	O	O

84	NN	O	O
and	NN	O	O
2	NN	O	O
.	NN	O	O

9	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

To	NN	O	O
confirm	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
a	NN	O	O
large	NN	O	O
linkage	NN	O	O
study	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
twelve	NN	O	O
additional	NN	O	O
families	NN	O	O
segregating	NN	O	O
for	NN	O	O
Alstrom	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
maximum	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
7	NN	O	O
.	NN	O	O

13	NN	O	O
(	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
00	NN	O	O
)	NN	O	O
for	NN	O	O
marker	NN	O	O
D2S2110	NN	O	O
and	NN	O	O
a	NN	O	O
maximum	NN	O	O
cumulative	NN	O	O
multipoint	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
9	NN	O	O
.	NN	O	O

16	NN	O	O
for	NN	O	O
marker	NN	O	O
D2S2110	NN	O	O
were	NN	O	O
observed	NN	O	O
,	NN	O	O
further	NN	O	O
supporting	NN	O	O
linkage	NN	O	O
to	NN	O	O
chromosome	NN	O	O
2p13	NN	O	O
.	NN	O	O

No	NN	O	O
evidence	NN	O	O
of	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
.	NN	O	O

Meiotic	NN	O	O
recombination	NN	O	O
events	NN	O	O
have	NN	O	O
localized	NN	O	O
the	NN	O	O
critical	NN	O	O
region	NN	O	O
containing	NN	O	O
ALMS1	NN	O	O
to	NN	O	O
a	NN	O	O
6	NN	O	O
.	NN	O	O

1	NN	O	O
-	NN	O	O
cM	NN	O	O
interval	NN	O	O
flanked	NN	O	O
by	NN	O	O
markers	NN	O	O
D2S327	NN	O	O
and	NN	O	O
D2S286	NN	O	O
.	NN	O	O

A	NN	O	O
fine	NN	O	O
resolution	NN	O	O
radiation	NN	O	O
hybrid	NN	O	O
map	NN	O	O
of	NN	O	O
31	NN	O	O
genes	NN	O	O
and	NN	O	O
markers	NN	O	O
has	NN	O	O
been	NN	O	O
constructed	NN	O	O
.	NN	O	O

Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
phenotypic	NN	O	O
variability	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
carrying	NN	O	O
the	NN	O	O
same	NN	O	O
PDS	NN	O	B-Disease
missense	NN	O	O
mutation	NN	O	O
.	NN	O	O

Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
comprises	NN	O	O
congenital	NN	O	B-Disease
sensorineural	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
,	NN	O	O
thyroid	NN	O	B-Disease
goiter	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
positive	NN	O	O
perchlorate	NN	O	O
discharge	NN	O	O
test	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
this	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
an	NN	O	O
anion	NN	O	O
transporter	NN	O	O
called	NN	O	O
pendrin	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
two	NN	O	O
consanguineous	NN	O	O
large	NN	O	O
families	NN	O	O
from	NN	O	O
Southern	NN	O	O
Tunisia	NN	O	O
comprising	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
23	NN	O	O
individuals	NN	O	O
affected	NN	O	O
with	NN	O	O
profound	NN	O	O
congenital	NN	O	B-Disease
deafness	NN	O	I-Disease
;	NN	O	O
the	NN	O	O
same	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
L445W	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
affected	NN	O	O
individuals	NN	O	O
.	NN	O	O

A	NN	O	O
widened	NN	O	B-Disease
vestibular	NN	O	I-Disease
aqueduct	NN	O	I-Disease
was	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
who	NN	O	O
underwent	NN	O	O
computed	NN	O	O
tomography	NN	O	O
(	NN	O	O
CT	NN	O	O
)	NN	O	O
scan	NN	O	O
exploration	NN	O	O
of	NN	O	O
the	NN	O	O
inner	NN	O	O
ear	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
goiter	NN	O	B-Disease
was	NN	O	O
present	NN	O	O
in	NN	O	O
only	NN	O	O
11	NN	O	O
affected	NN	O	O
individuals	NN	O	O
,	NN	O	O
who	NN	O	O
interestingly	NN	O	O
had	NN	O	O
a	NN	O	O
normal	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
perchlorate	NN	O	O
discharge	NN	O	O
test	NN	O	O
whenever	NN	O	O
performed	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
results	NN	O	O
question	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
perchlorate	NN	O	O
test	NN	O	O
for	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
support	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
the	NN	O	O
assessment	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
severe	NN	O	O
to	NN	O	O
profound	NN	O	O
congenital	NN	O	B-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
inner	NN	O	B-Disease
ear	NN	O	I-Disease
morphological	NN	O	I-Disease
anomaly	NN	O	I-Disease
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
thyroid	NN	O	B-Disease
goiter	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Knobloch	NN	O	B-Disease
syndrome	NN	O	I-Disease
involving	NN	O	O
midline	NN	O	B-Disease
scalp	NN	O	I-Disease
defect	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
frontal	NN	O	I-Disease
region	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
4	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
boy	NN	O	O
with	NN	O	O
Knobloch	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

He	NN	O	O
has	NN	O	O
vitreoretinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
,	NN	O	O
high	NN	O	B-Disease
myopia	NN	O	I-Disease
,	NN	O	O
cataract	NN	O	B-Disease
,	NN	O	O
telecanthus	NN	O	B-Disease
,	NN	O	O
hypertelorism	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
a	NN	O	O
high	NN	O	B-Disease
-	NN	O	I-Disease
arched	NN	O	I-Disease
palate	NN	O	I-Disease
.	NN	O	O

He	NN	O	O
also	NN	O	O
has	NN	O	O
a	NN	O	O
defect	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
anterior	NN	O	I-Disease
midline	NN	O	I-Disease
scalp	NN	O	I-Disease
with	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
frontal	NN	O	O
bone	NN	O	O
as	NN	O	O
documented	NN	O	O
by	NN	O	O
a	NN	O	O
computed	NN	O	O
tomography	NN	O	O
(	NN	O	O
CT	NN	O	O
)	NN	O	O
scan	NN	O	O
.	NN	O	O

The	NN	O	O
brain	NN	O	O
was	NN	O	O
normal	NN	O	O
on	NN	O	O
CT	NN	O	O
scan	NN	O	O
and	NN	O	O
magnetic	NN	O	O
resonance	NN	O	O
imaging	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
review	NN	O	O
of	NN	O	O
the	NN	O	O
23	NN	O	O
published	NN	O	O
cases	NN	O	O
with	NN	O	O
this	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Our	NN	O	O
patient	NN	O	O
illustrates	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
investigating	NN	O	O
for	NN	O	O
underlying	NN	O	O
ocular	NN	O	O
and	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
pathology	NN	O	O
whenever	NN	O	O
midline	NN	O	B-Disease
scalp	NN	O	I-Disease
defects	NN	O	I-Disease
are	NN	O	O
present	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
double	NN	O	O
-	NN	O	O
strand	NN	O	O
break	NN	O	O
repair	NN	O	O
gene	NN	O	O
hMRE11	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
an	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
-	NN	O	I-Disease
like	NN	O	I-Disease
disorder	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
hypomorphic	NN	O	O
mutations	NN	O	O
in	NN	O	O
hMRE11	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
ATM	NN	O	O
,	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
certain	NN	O	O
individuals	NN	O	O
with	NN	O	O
an	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
-	NN	O	I-Disease
like	NN	O	I-Disease
disorder	NN	O	I-Disease
(	NN	O	O
ATLD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
cellular	NN	O	O
features	NN	O	O
resulting	NN	O	O
from	NN	O	O
these	NN	O	O
hMRE11	NN	O	O
mutations	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
seen	NN	O	O
in	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
as	NN	O	O
well	NN	O	O
as	NN	O	O
NBS	NN	O	B-Disease
and	NN	O	O
include	NN	O	O
hypersensitivity	NN	O	B-Disease
to	NN	O	I-Disease
ionizing	NN	O	I-Disease
radiation	NN	O	I-Disease
,	NN	O	O
radioresistant	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
abrogation	NN	O	O
of	NN	O	O
ATM	NN	O	O
-	NN	O	O
dependent	NN	O	O
events	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Jun	NN	O	O
kinase	NN	O	O
following	NN	O	O
exposure	NN	O	O
to	NN	O	O
gamma	NN	O	O
irradiation	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mutant	NN	O	O
hMre11	NN	O	O
proteins	NN	O	O
retain	NN	O	O
some	NN	O	O
ability	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
hRad50	NN	O	O
and	NN	O	O
Nbs1	NN	O	O
,	NN	O	O
formation	NN	O	O
of	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
-	NN	O	O
induced	NN	O	O
hMre11	NN	O	O
and	NN	O	O
Nbs1	NN	O	O
foci	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
hMRE11	NN	O	O
mutant	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
ATM	NN	O	O
and	NN	O	O
the	NN	O	O
hMre11	NN	O	O
/	NN	O	O
hRad50	NN	O	O
/	NN	O	O
Nbs1	NN	O	O
protein	NN	O	O
complex	NN	O	O
act	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
DNA	NN	O	O
damage	NN	O	O
response	NN	O	O
pathway	NN	O	O
and	NN	O	O
link	NN	O	O
hMre11	NN	O	O
to	NN	O	O
the	NN	O	O
complex	NN	O	O
pathology	NN	O	O
of	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
TNFRSF11A	NN	O	O
,	NN	O	O
affecting	NN	O	O
the	NN	O	O
signal	NN	O	O
peptide	NN	O	O
of	NN	O	O
RANK	NN	O	O
,	NN	O	O
cause	NN	O	O
familial	NN	O	B-Disease
expansile	NN	O	I-Disease
osteolysis	NN	O	I-Disease
.	NN	O	O

Familial	NN	O	B-Disease
expansile	NN	O	I-Disease
osteolysis	NN	O	I-Disease
(	NN	O	O
FEO	NN	O	B-Disease
,	NN	O	O
MIM	NN	O	O
174810	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
,	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
bone	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
focal	NN	O	O
areas	NN	O	O
of	NN	O	O
increased	NN	O	B-Disease
bone	NN	O	I-Disease
remodelling	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
osteolytic	NN	O	B-Disease
lesions	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
develop	NN	O	O
usually	NN	O	O
in	NN	O	O
the	NN	O	O
long	NN	O	O
bones	NN	O	O
during	NN	O	O
early	NN	O	O
adulthood	NN	O	O
,	NN	O	O
show	NN	O	O
increased	NN	O	O
osteoblast	NN	O	O
and	NN	O	O
osteoclast	NN	O	O
activity	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
linkage	NN	O	O
studies	NN	O	O
mapped	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
FEO	NN	O	B-Disease
to	NN	O	O
an	NN	O	O
interval	NN	O	O
of	NN	O	O
less	NN	O	O
than	NN	O	O
5	NN	O	O
cM	NN	O	O
between	NN	O	O
D18S64	NN	O	O
and	NN	O	O
D18S51	NN	O	O
on	NN	O	O
chromosome	NN	O	O
18q21	NN	O	O
.	NN	O	O

2	NN	O	O
-	NN	O	O
21	NN	O	O
2	NN	O	O
-	NN	O	O
21	NN	O	O
.	NN	O	O

3	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
Northern	NN	O	O
Irish	NN	O	O
family	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
receptor	NN	O	O
activator	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
-	NN	O	O
kappa	NN	O	O
B	NN	O	O
(	NN	O	O
RANK	NN	O	O
;	NN	O	O
ref	NN	O	O
.	NN	O	O
5	NN	O	O
)	NN	O	O
,	NN	O	O
TNFRSF11A	NN	O	O
,	NN	O	O
maps	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

RANK	NN	O	O
is	NN	O	O
essential	NN	O	O
in	NN	O	O
osteoclast	NN	O	O
formation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
two	NN	O	O
heterozygous	NN	O	O
insertion	NN	O	O
mutations	NN	O	O
in	NN	O	O
exon	NN	O	O
1	NN	O	O
of	NN	O	O
TNFRSF11A	NN	O	O
in	NN	O	O
affected	NN	O	O
members	NN	O	O
of	NN	O	O
four	NN	O	O
families	NN	O	O
with	NN	O	O
FEO	NN	O	B-Disease
or	NN	O	O
familial	NN	O	B-Disease
Paget	NN	O	I-Disease
disease	NN	O	I-Disease
of	NN	O	I-Disease
bone	NN	O	I-Disease
(	NN	O	O
PDB	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

One	NN	O	O
was	NN	O	O
a	NN	O	O
duplication	NN	O	O
of	NN	O	O
18	NN	O	O
bases	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
a	NN	O	O
duplication	NN	O	O
of	NN	O	O
27	NN	O	O
bases	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
affected	NN	O	O
the	NN	O	O
signal	NN	O	O
peptide	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
RANK	NN	O	O
molecule	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
recombinant	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
RANK	NN	O	O
proteins	NN	O	O
revealed	NN	O	O
perturbations	NN	O	O
in	NN	O	O
expression	NN	O	O
levels	NN	O	O
and	NN	O	O
lack	NN	O	O
of	NN	O	O
normal	NN	O	O
cleavage	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
peptide	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
RANK	NN	O	O
-	NN	O	O
mediated	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
-	NN	O	O
kappaB	NN	O	O
(	NN	O	O
NF	NN	O	O
-	NN	O	O
kappaB	NN	O	O
)	NN	O	O
signalling	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
activating	NN	O	O
mutation	NN	O	O
.	NN	O	O

Cardiac	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
channel	NN	O	O
dysfunction	NN	O	O
in	NN	O	O
Brugada	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
aggravated	NN	O	O
by	NN	O	O
beta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
subunit	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
human	NN	O	O
cardiac	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
channel	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
(	NN	O	O
hH1	NN	O	O
)	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
chromosome	NN	O	O
3	NN	O	O
-	NN	O	O
linked	NN	O	O
congenital	NN	O	B-Disease
long	NN	O	I-Disease
-	NN	O	I-Disease
QT	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
LQT3	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
idiopathic	NN	O	B-Disease
ventricular	NN	O	I-Disease
fibrillation	NN	O	I-Disease
(	NN	O	O
IVF	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

An	NN	O	O
auxiliary	NN	O	O
beta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
subunit	NN	O	O
,	NN	O	O
widely	NN	O	O
expressed	NN	O	O
in	NN	O	O
excitable	NN	O	O
tissues	NN	O	O
,	NN	O	O
shifts	NN	O	O
the	NN	O	O
voltage	NN	O	O
dependence	NN	O	O
of	NN	O	O
steady	NN	O	O
-	NN	O	O
state	NN	O	O
inactivation	NN	O	O
toward	NN	O	O
more	NN	O	O
negative	NN	O	O
potentials	NN	O	O
and	NN	O	O
restores	NN	O	O
normal	NN	O	O
gating	NN	O	O
kinetics	NN	O	O
of	NN	O	O
brain	NN	O	O
and	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
channels	NN	O	O
expressed	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
oocytes	NN	O	O
but	NN	O	O
has	NN	O	O
little	NN	O	O
if	NN	O	O
any	NN	O	O
functional	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
cardiac	NN	O	O
isoform	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
characterize	NN	O	O
the	NN	O	O
altered	NN	O	O
effects	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	O
beta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
subunit	NN	O	O
(	NN	O	O
hbeta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
heterologously	NN	O	O
expressed	NN	O	O
hH1	NN	O	O
mutation	NN	O	O
(	NN	O	O
T1620M	NN	O	O
)	NN	O	O
previously	NN	O	O
associated	NN	O	O
with	NN	O	O
IVF	NN	O	B-Disease
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
When	NN	O	O
expressed	NN	O	O
alone	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
oocytes	NN	O	O
,	NN	O	O
T1620M	NN	O	O
exhibited	NN	O	O
no	NN	O	O
persistent	NN	O	O
currents	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
LQT3	NN	O	B-Disease
mutant	NN	O	O
channels	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
midpoint	NN	O	O
of	NN	O	O
steady	NN	O	O
-	NN	O	O
state	NN	O	O
inactivation	NN	O	O
(	NN	O	O
V	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
was	NN	O	O
significantly	NN	O	O
shifted	NN	O	O
toward	NN	O	O
more	NN	O	O
positive	NN	O	O
potentials	NN	O	O
than	NN	O	O
for	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
hH1	NN	O	O
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
hbeta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
alter	NN	O	O
current	NN	O	O
decay	NN	O	O
or	NN	O	O
recovery	NN	O	O
from	NN	O	O
inactivation	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
hH1	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
further	NN	O	O
shifted	NN	O	O
the	NN	O	O
V	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
accelerated	NN	O	O
the	NN	O	O
recovery	NN	O	O
from	NN	O	O
inactivation	NN	O	O
of	NN	O	O
T1620M	NN	O	O
.	NN	O	O

Oocyte	NN	O	O
macropatch	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
kinetics	NN	O	O
of	NN	O	O
T1620M	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
coexpression	NN	O	O
of	NN	O	O
hbeta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
exposes	NN	O	O
a	NN	O	O
more	NN	O	O
severe	NN	O	O
functional	NN	O	O
defect	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
greater	NN	O	O
overlap	NN	O	O
in	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
channel	NN	O	O
inactivation	NN	O	O
and	NN	O	O
activation	NN	O	O
(	NN	O	O
window	NN	O	O
current	NN	O	O
)	NN	O	O
in	NN	O	O
T1620M	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
proposed	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
pathophysiological	NN	O	O
mechanism	NN	O	O
of	NN	O	O
IVF	NN	O	B-Disease
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

One	NN	O	O
possible	NN	O	O
explanation	NN	O	O
for	NN	O	O
our	NN	O	O
finding	NN	O	O
is	NN	O	O
an	NN	O	O
altered	NN	O	O
alpha	NN	O	O
-	NN	O	O
/	NN	O	O
beta	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
subunit	NN	O	O
association	NN	O	O
in	NN	O	O
the	NN	O	O
mutant	NN	O	O
.	NN	O	O
.	NN	O	O

Meiotic	NN	O	O
segregation	NN	O	O
analysis	NN	O	O
of	NN	O	O
RB1	NN	O	O
alleles	NN	O	O
in	NN	O	O
retinoblastoma	NN	O	B-Disease
pedigrees	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
single	NN	O	O
-	NN	O	O
sperm	NN	O	O
typing	NN	O	O
.	NN	O	O

In	NN	O	O
hereditary	NN	O	B-Disease
retinoblastoma	NN	O	I-Disease
,	NN	O	O
different	NN	O	O
epidemiological	NN	O	O
studies	NN	O	O
have	NN	O	O
indicated	NN	O	O
a	NN	O	O
preferential	NN	O	O
paternal	NN	O	O
transmission	NN	O	O
of	NN	O	O
mutant	NN	O	O
retinoblastoma	NN	O	B-Disease
alleles	NN	O	O
to	NN	O	O
offspring	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
a	NN	O	O
meiotic	NN	O	O
drive	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
this	NN	O	O
mechanism	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
sperm	NN	O	O
samples	NN	O	O
from	NN	O	O
six	NN	O	O
individuals	NN	O	O
from	NN	O	O
five	NN	O	O
unrelated	NN	O	O
families	NN	O	O
affected	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
retinoblastoma	NN	O	I-Disease
.	NN	O	O

Single	NN	O	O
-	NN	O	O
sperm	NN	O	O
typing	NN	O	O
techniques	NN	O	O
were	NN	O	O
performed	NN	O	O
for	NN	O	O
each	NN	O	O
sample	NN	O	O
by	NN	O	O
study	NN	O	O
of	NN	O	O
two	NN	O	O
informative	NN	O	O
short	NN	O	O
tandem	NN	O	O
repeats	NN	O	O
located	NN	O	O
either	NN	O	O
in	NN	O	O
or	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
RB1	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
segregation	NN	O	O
probability	NN	O	O
of	NN	O	O
mutant	NN	O	O
RB1	NN	O	O
alleles	NN	O	O
in	NN	O	O
sperm	NN	O	O
samples	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
SPERMSEG	NN	O	O
program	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
experimental	NN	O	O
parameters	NN	O	O
,	NN	O	O
recombination	NN	O	O
fractions	NN	O	O
between	NN	O	O
the	NN	O	O
markers	NN	O	O
,	NN	O	O
and	NN	O	O
segregation	NN	O	O
parameters	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
952	NN	O	O
single	NN	O	O
sperm	NN	O	O
from	NN	O	O
the	NN	O	O
six	NN	O	O
donors	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

We	NN	O	O
detected	NN	O	O
a	NN	O	O
significant	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
in	NN	O	O
the	NN	O	O
data	NN	O	O
as	NN	O	O
a	NN	O	O
whole	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
0099	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
the	NN	O	O
segregation	NN	O	O
rate	NN	O	O
across	NN	O	O
donors	NN	O	O
(	NN	O	O
.	NN	O	O
0092	NN	O	O
)	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
shows	NN	O	O
that	NN	O	O
this	NN	O	O
result	NN	O	O
can	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
in	NN	O	O
favor	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
allele	NN	O	O
in	NN	O	O
one	NN	O	O
donor	NN	O	O
only	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
provide	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
significant	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
donors	NN	O	O
.	NN	O	O

The	NN	O	O
segregation	NN	O	O
distortion	NN	O	O
favoring	NN	O	O
the	NN	O	O
mutant	NN	O	O
RB1	NN	O	O
allele	NN	O	O
does	NN	O	O
not	NN	O	O
seem	NN	O	O
to	NN	O	O
occur	NN	O	O
during	NN	O	O
spermatogenesis	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
thus	NN	O	O
,	NN	O	O
meiotic	NN	O	O
drive	NN	O	O
may	NN	O	O
result	NN	O	O
either	NN	O	O
from	NN	O	O
various	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
fertilization	NN	O	O
advantage	NN	O	O
or	NN	O	O
a	NN	O	O
better	NN	O	O
mobility	NN	O	O
in	NN	O	O
sperm	NN	O	O
bearing	NN	O	O
a	NN	O	O
mutant	NN	O	O
RB1	NN	O	O
gene	NN	O	O
,	NN	O	O
or	NN	O	O
from	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
defectively	NN	O	O
imprinted	NN	O	O
gene	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
:	NN	O	O
an	NN	O	O
overview	NN	O	O
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
,	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
neurodegenerative	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
of	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
ataxias	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
recent	NN	O	O
discovery	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
this	NN	O	O
condition	NN	O	O
,	NN	O	O
FRDA	NN	O	O
,	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
rapid	NN	O	O
advances	NN	O	O
in	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

About	NN	O	O
98	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
have	NN	O	O
an	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
GAA	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
intron	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
leads	NN	O	O
to	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
,	NN	O	O
frataxin	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
mounting	NN	O	O
evidence	NN	O	O
to	NN	O	O
suggest	NN	O	O
that	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
accumulation	NN	O	O
of	NN	O	O
iron	NN	O	O
in	NN	O	O
mitochondria	NN	O	O
leading	NN	O	O
to	NN	O	O
excess	NN	O	O
production	NN	O	O
of	NN	O	O
free	NN	O	O
radicals	NN	O	O
,	NN	O	O
which	NN	O	O
then	NN	O	O
results	NN	O	O
in	NN	O	O
cellular	NN	O	O
damage	NN	O	O
and	NN	O	O
death	NN	O	O
.	NN	O	O

Currently	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
known	NN	O	O
treatment	NN	O	O
that	NN	O	O
alters	NN	O	O
the	NN	O	O
natural	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
discovery	NN	O	O
of	NN	O	O
the	NN	O	O
FRDA	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
its	NN	O	O
possible	NN	O	O
function	NN	O	O
has	NN	O	O
raised	NN	O	O
hope	NN	O	O
that	NN	O	O
rational	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
will	NN	O	O
be	NN	O	O
developed	NN	O	O
.	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
retinoschisis	NN	O	I-Disease
with	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
retinoschisis	NN	O	I-Disease
(	NN	O	O
XLRS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
relatively	NN	O	O
rare	NN	O	O
vitreoretinal	NN	O	B-Disease
dystrophy	NN	O	I-Disease
that	NN	O	O
causes	NN	O	O
visual	NN	O	B-Disease
loss	NN	O	I-Disease
in	NN	O	O
young	NN	O	O
men	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
disease	NN	O	O
,	NN	O	O
designated	NN	O	O
XLRS1	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
,	NN	O	O
and	NN	O	O
several	NN	O	O
deleterious	NN	O	O
gene	NN	O	O
mutations	NN	O	O
were	NN	O	O
reported	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
To	NN	O	O
analyze	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
clinically	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
XLRS	NN	O	B-Disease
formutational	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
.	NN	O	O

METHODS	NN	O	O
Ten	NN	O	O
patients	NN	O	O
with	NN	O	O
XLRS	NN	O	B-Disease
underwent	NN	O	O
full	NN	O	O
ophthalmologic	NN	O	O
examination	NN	O	O
,	NN	O	O
including	NN	O	O
slitlamp	NN	O	O
biomicroscopy	NN	O	O
and	NN	O	O
dilated	NN	O	O
funduscopy	NN	O	O
.	NN	O	O

Genomic	NN	O	O
DNA	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
leukocytes	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
were	NN	O	O
amplified	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
analyzed	NN	O	O
using	NN	O	O
a	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
method	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
10	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
were	NN	O	O
identical	NN	O	O
in	NN	O	O
each	NN	O	O
of	NN	O	O
2	NN	O	O
pairs	NN	O	O
of	NN	O	O
brothers	NN	O	O
.	NN	O	O

Six	NN	O	O
of	NN	O	O
the	NN	O	O
point	NN	O	O
mutations	NN	O	O
represented	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
1	NN	O	O
was	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
was	NN	O	O
a	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
.	NN	O	O

Five	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
are	NN	O	O
newly	NN	O	O
reported	NN	O	O
herein	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
The	NN	O	O
discovery	NN	O	O
of	NN	O	O
new	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
increases	NN	O	O
the	NN	O	O
available	NN	O	O
information	NN	O	O
regarding	NN	O	O
the	NN	O	O
spectrum	NN	O	O
of	NN	O	O
genetic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
and	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
XLRS	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
limited	NN	O	O
data	NN	O	O
failed	NN	O	O
to	NN	O	O
reveal	NN	O	O
a	NN	O	O
correlation	NN	O	O
between	NN	O	O
mutation	NN	O	O
and	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
.	NN	O	O

CLINICAL	NN	O	O
RELEVANCE	NN	O	O
Identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
XLRS1	NN	O	O
gene	NN	O	O
and	NN	O	O
expanded	NN	O	O
information	NN	O	O
on	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
will	NN	O	O
facilitate	NN	O	O
early	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
appropriate	NN	O	O
early	NN	O	O
therapy	NN	O	O
,	NN	O	O
and	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
regarding	NN	O	O
the	NN	O	O
prognosis	NN	O	O
of	NN	O	O
XLRS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Atm	NN	O	O
and	NN	O	O
Bax	NN	O	O
cooperate	NN	O	O
in	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
-	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

Ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
hereditary	NN	O	B-Disease
multisystemic	NN	O	I-Disease
disease	NN	O	I-Disease
resulting	NN	O	O
from	NN	O	O
mutations	NN	O	O
of	NN	O	O
ataxia	NN	O	B-Disease
telangiectasia	NN	O	I-Disease
,	NN	O	O
mutated	NN	O	O
(	NN	O	O
ATM	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
neurodegeneration	NN	O	B-Disease
,	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
immune	NN	O	B-Disease
defects	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hypersensitivity	NN	O	B-Disease
to	NN	O	I-Disease
ionizing	NN	O	I-Disease
radiation	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
details	NN	O	O
of	NN	O	O
ATM	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
nervous	NN	O	O
system	NN	O	O
are	NN	O	O
unclear	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
neurological	NN	O	B-Disease
lesion	NN	O	I-Disease
in	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
becomes	NN	O	O
apparent	NN	O	O
early	NN	O	O
in	NN	O	O
life	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
developmental	NN	O	O
origin	NN	O	O
.	NN	O	O

The	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
(	NN	O	O
CNS	NN	O	O
)	NN	O	O
of	NN	O	O
Atm	NN	O	O
-	NN	O	O
null	NN	O	O
mice	NN	O	O
shows	NN	O	O
a	NN	O	O
pronounced	NN	O	O
defect	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
genotoxic	NN	O	O
stress	NN	O	O
,	NN	O	O
suggesting	NN	O	O
ATM	NN	O	O
functions	NN	O	O
to	NN	O	O
eliminate	NN	O	O
neurons	NN	O	O
with	NN	O	O
excessive	NN	O	O
genomic	NN	O	O
damage	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
death	NN	O	O
effector	NN	O	O
Bax	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
a	NN	O	O
large	NN	O	O
proportion	NN	O	O
of	NN	O	O
Atm	NN	O	O
-	NN	O	O
dependent	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
developing	NN	O	O
CNS	NN	O	O
after	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
(	NN	O	O
IR	NN	O	O
)	NN	O	O
.	NN	O	O

Although	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
CNS	NN	O	O
in	NN	O	O
both	NN	O	O
Bax	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
and	NN	O	O
Atm	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
were	NN	O	O
radioresistant	NN	O	O
,	NN	O	O
mice	NN	O	O
nullizygous	NN	O	O
for	NN	O	O
both	NN	O	O
Bax	NN	O	O
and	NN	O	O
Atm	NN	O	O
showed	NN	O	O
additional	NN	O	O
reduction	NN	O	O
in	NN	O	O
IR	NN	O	O
-	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
CNS	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
major	NN	O	O
IR	NN	O	O
-	NN	O	O
induced	NN	O	O
apoptotic	NN	O	O
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
CNS	NN	O	O
requires	NN	O	O
Atm	NN	O	O
and	NN	O	O
Bax	NN	O	O
,	NN	O	O
a	NN	O	O
p53	NN	O	O
-	NN	O	O
dependent	NN	O	O
collateral	NN	O	O
pathway	NN	O	O
exists	NN	O	O
that	NN	O	O
has	NN	O	O
both	NN	O	O
Atm	NN	O	O
-	NN	O	O
and	NN	O	O
Bax	NN	O	O
-	NN	O	O
independent	NN	O	O
branches	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
Atm	NN	O	O
-	NN	O	O
and	NN	O	O
Bax	NN	O	O
-	NN	O	O
dependent	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
CNS	NN	O	O
also	NN	O	O
required	NN	O	O
caspase	NN	O	O
-	NN	O	O
3	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
implicate	NN	O	O
Bax	NN	O	O
and	NN	O	O
caspase	NN	O	O
-	NN	O	O
3	NN	O	O
as	NN	O	O
death	NN	O	O
effectors	NN	O	O
in	NN	O	O
neurodegenerative	NN	O	O
pathways	NN	O	O
.	NN	O	O
.	NN	O	O

Haim	NN	O	B-Disease
-	NN	O	I-Disease
Munk	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
Papillon	NN	O	B-Disease
-	NN	O	I-Disease
Lefevre	NN	O	I-Disease
syndrome	NN	O	I-Disease
are	NN	O	O
allelic	NN	O	O
mutations	NN	O	O
in	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
many	NN	O	O
palmoplantar	NN	O	B-Disease
keratoderma	NN	O	I-Disease
(	NN	O	I-Disease
PPK	NN	O	I-Disease
)	NN	O	I-Disease
conditions	NN	O	I-Disease
,	NN	O	O
only	NN	O	O
Papillon	NN	O	B-Disease
-	NN	O	I-Disease
Lefevre	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PLS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Haim	NN	O	B-Disease
-	NN	O	I-Disease
Munk	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
HMS	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
premature	NN	O	O
periodontal	NN	O	O
destruction	NN	O	O
.	NN	O	O

Although	NN	O	O
both	NN	O	O
PLS	NN	O	B-Disease
and	NN	O	O
HMS	NN	O	B-Disease
share	NN	O	O
the	NN	O	O
cardinal	NN	O	O
features	NN	O	O
of	NN	O	O
PPK	NN	O	B-Disease
and	NN	O	O
severe	NN	O	O
periodontitis	NN	O	B-Disease
,	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
additional	NN	O	O
findings	NN	O	O
are	NN	O	O
reported	NN	O	O
in	NN	O	O
HMS	NN	O	B-Disease
including	NN	O	O
arachnodactyly	NN	O	B-Disease
,	NN	O	O
acro	NN	O	B-Disease
-	NN	O	I-Disease
osteolysis	NN	O	I-Disease
,	NN	O	O
atrophic	NN	O	B-Disease
changes	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
nails	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
a	NN	O	O
radiographic	NN	O	B-Disease
deformity	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fingers	NN	O	I-Disease
.	NN	O	O

While	NN	O	O
PLS	NN	O	B-Disease
cases	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
throughout	NN	O	O
the	NN	O	O
world	NN	O	O
,	NN	O	O
HMS	NN	O	B-Disease
has	NN	O	O
only	NN	O	O
been	NN	O	O
described	NN	O	O
among	NN	O	O
descendants	NN	O	O
of	NN	O	O
a	NN	O	O
religious	NN	O	O
isolate	NN	O	O
originally	NN	O	O
from	NN	O	O
Cochin	NN	O	O
,	NN	O	O
India	NN	O	O
.	NN	O	O

Parental	NN	O	O
consanguinity	NN	O	O
is	NN	O	O
a	NN	O	O
characteristic	NN	O	O
of	NN	O	O
many	NN	O	O
cases	NN	O	O
of	NN	O	O
both	NN	O	O
conditions	NN	O	O
.	NN	O	O

Although	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
transmission	NN	O	O
of	NN	O	O
PLS	NN	O	B-Disease
is	NN	O	O
evident	NN	O	O
,	NN	O	O
a	NN	O	O
more	NN	O	O
"	NN	O	O
complex	NN	O	O
"	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
pattern	NN	O	O
of	NN	O	O
inheritance	NN	O	O
with	NN	O	O
phenotypic	NN	O	O
influences	NN	O	O
from	NN	O	O
a	NN	O	O
closely	NN	O	O
linked	NN	O	O
modifying	NN	O	O
locus	NN	O	O
has	NN	O	O
been	NN	O	O
hypothesised	NN	O	O
for	NN	O	O
HMS	NN	O	B-Disease
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
underlying	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
in	NN	O	O
PLS	NN	O	B-Disease
.	NN	O	O

To	NN	O	O
determine	NN	O	O
if	NN	O	O
a	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
mutation	NN	O	O
is	NN	O	O
also	NN	O	O
responsible	NN	O	O
for	NN	O	O
HMS	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
sequenced	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
affected	NN	O	O
and	NN	O	O
unaffected	NN	O	O
subjects	NN	O	O
from	NN	O	O
the	NN	O	O
Cochin	NN	O	O
isolate	NN	O	O
in	NN	O	O
which	NN	O	O
both	NN	O	O
the	NN	O	O
PLS	NN	O	B-Disease
and	NN	O	O
HMS	NN	O	B-Disease
phenotypes	NN	O	O
appear	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
(	NN	O	O
exon	NN	O	O
6	NN	O	O
,	NN	O	O
2127A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
that	NN	O	O
changes	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
amino	NN	O	O
acid	NN	O	O
in	NN	O	O
the	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
peptide	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
segregates	NN	O	O
with	NN	O	O
HMS	NN	O	B-Disease
in	NN	O	O
four	NN	O	O
nuclear	NN	O	O
families	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
shared	NN	O	O
common	NN	O	O
haplotype	NN	O	O
for	NN	O	O
genetic	NN	O	O
loci	NN	O	O
flanking	NN	O	O
the	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
suggests	NN	O	O
that	NN	O	O
affected	NN	O	O
subjects	NN	O	O
descended	NN	O	O
from	NN	O	O
the	NN	O	O
Cochin	NN	O	O
isolate	NN	O	O
are	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
mutation	NN	O	O
inherited	NN	O	O
"	NN	O	O
identical	NN	O	O
by	NN	O	O
descent	NN	O	O
"	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
ancestor	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
supports	NN	O	O
simple	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
inheritance	NN	O	O
for	NN	O	O
HMS	NN	O	B-Disease
in	NN	O	O
these	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
report	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
exon	NN	O	O
6	NN	O	O
CTSC	NN	O	O
codon	NN	O	O
(	NN	O	O
2126C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
Turkish	NN	O	O
family	NN	O	O
with	NN	O	O
classical	NN	O	O
PLS	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
PLS	NN	O	B-Disease
and	NN	O	O
HMS	NN	O	B-Disease
are	NN	O	O
allelic	NN	O	O
variants	NN	O	O
of	NN	O	O
cathepsin	NN	O	O
C	NN	O	O
gene	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

ATM	NN	O	O
-	NN	O	O
heterozygous	NN	O	O
germline	NN	O	O
mutations	NN	O	O
contribute	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
-	NN	O	O
susceptibility	NN	O	O
.	NN	O	O

Approximately	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
-	NN	O	O
1	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
has	NN	O	O
been	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
(	NN	O	O
MIM	NN	O	O
208900	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
ATM	NN	O	O
-	NN	O	O
heterozygotes	NN	O	O
have	NN	O	O
an	NN	O	O
increased	NN	O	O
relative	NN	O	O
risk	NN	O	O
for	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
was	NN	O	O
supported	NN	O	O
by	NN	O	O
some	NN	O	O
studies	NN	O	O
but	NN	O	O
not	NN	O	O
confirmed	NN	O	O
by	NN	O	O
others	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
this	NN	O	O
discrepancy	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
ATM	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
selected	NN	O	O
group	NN	O	O
of	NN	O	O
Dutch	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
ATM	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
normal	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
using	NN	O	O
the	NN	O	O
protein	NN	O	O
-	NN	O	O
truncation	NN	O	O
test	NN	O	O
followed	NN	O	O
by	NN	O	O
genomic	NN	O	O
-	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
.	NN	O	O

A	NN	O	O
high	NN	O	O
percentage	NN	O	O
of	NN	O	O
ATM	NN	O	O
germline	NN	O	O
mutations	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
sporadic	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
82	NN	O	O
patients	NN	O	O
included	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
had	NN	O	O
developed	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
at	NN	O	O
age	NN	O	O
<	NN	O	O
45	NN	O	O
and	NN	O	O
had	NN	O	O
survived	NN	O	O
>	NN	O	O
/	NN	O	O
=	NN	O	O
5	NN	O	O
years	NN	O	O
(	NN	O	O
mean	NN	O	O
15	NN	O	O
years	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
33	NN	O	O
(	NN	O	O
40	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
a	NN	O	O
contralateral	NN	O	O
breast	NN	O	O
tumor	NN	O	O
had	NN	O	O
been	NN	O	O
diagnosed	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
patients	NN	O	O
we	NN	O	O
identified	NN	O	O
seven	NN	O	O
(	NN	O	O
8	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
ATM	NN	O	O
germline	NN	O	O
mutations	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
five	NN	O	O
are	NN	O	O
distinct	NN	O	O
.	NN	O	O

One	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
(	NN	O	O
IVS10	NN	O	O
-	NN	O	O
6T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
was	NN	O	O
detected	NN	O	O
three	NN	O	O
times	NN	O	O
in	NN	O	O
our	NN	O	O
series	NN	O	O
.	NN	O	O

Four	NN	O	O
heterozygous	NN	O	O
carriers	NN	O	O
were	NN	O	O
patients	NN	O	O
with	NN	O	O
bilateral	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
mutations	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
are	NN	O	O
"	NN	O	O
A	NN	O	O
-	NN	O	O
T	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
"	NN	O	O
mutations	NN	O	O
that	NN	O	O
might	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
in	NN	O	O
heterozygotes	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
ATM	NN	O	O
heterozygotes	NN	O	O
have	NN	O	O
an	NN	O	O
approximately	NN	O	O
ninefold	NN	O	O
-	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
developing	NN	O	O
a	NN	O	O
type	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
characterized	NN	O	O
by	NN	O	O
frequent	NN	O	O
bilateral	NN	O	O
occurrence	NN	O	O
,	NN	O	O
early	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
,	NN	O	O
and	NN	O	O
long	NN	O	O
-	NN	O	O
term	NN	O	O
survival	NN	O	O
.	NN	O	O

The	NN	O	O
specific	NN	O	O
characteristics	NN	O	O
of	NN	O	O
our	NN	O	O
population	NN	O	O
of	NN	O	O
patients	NN	O	O
may	NN	O	O
explain	NN	O	O
why	NN	O	O
such	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
other	NN	O	O
series	NN	O	O
.	NN	O	O

Human	NN	O	O
mutations	NN	O	O
in	NN	O	O
glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
reflect	NN	O	O
evolutionary	NN	O	O
history	NN	O	O
.	NN	O	O

Glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytosolic	NN	O	O
enzyme	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
housekeeping	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
gene	NN	O	O
whose	NN	O	O
main	NN	O	O
function	NN	O	O
is	NN	O	O
to	NN	O	O
produce	NN	O	O
NADPH	NN	O	O
,	NN	O	O
a	NN	O	O
key	NN	O	O
electron	NN	O	O
donor	NN	O	O
in	NN	O	O
the	NN	O	O
defense	NN	O	O
against	NN	O	O
oxidizing	NN	O	O
agents	NN	O	O
and	NN	O	O
in	NN	O	O
reductive	NN	O	O
biosynthetic	NN	O	O
reactions	NN	O	O
.	NN	O	O

Inherited	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
either	NN	O	O
episodic	NN	O	B-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
(	NN	O	O
triggered	NN	O	O
by	NN	O	O
fava	NN	O	O
beans	NN	O	O
or	NN	O	O
other	NN	O	O
agents	NN	O	O
)	NN	O	O
or	NN	O	O
life	NN	O	B-Disease
-	NN	O	I-Disease
long	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
an	NN	O	O
evolutionary	NN	O	O
analysis	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
to	NN	O	O
understanding	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
a	NN	O	O
housekeeping	NN	O	O
gene	NN	O	O
.	NN	O	O

From	NN	O	O
the	NN	O	O
alignment	NN	O	O
of	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
(	NN	O	O
aa	NN	O	O
)	NN	O	O
sequence	NN	O	O
of	NN	O	O
52	NN	O	O
glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
species	NN	O	O
from	NN	O	O
42	NN	O	O
different	NN	O	O
organisms	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
striking	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
aa	NN	O	O
replacements	NN	O	O
that	NN	O	O
cause	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
humans	NN	O	O
and	NN	O	O
the	NN	O	O
sequence	NN	O	O
conservation	NN	O	O
of	NN	O	O
G6PD	NN	O	O
two	NN	O	O
-	NN	O	O
thirds	NN	O	O
of	NN	O	O
such	NN	O	O
replacements	NN	O	O
are	NN	O	O
in	NN	O	O
highly	NN	O	O
and	NN	O	O
moderately	NN	O	O
conserved	NN	O	O
(	NN	O	O
50	NN	O	O
-	NN	O	O
99	NN	O	O
%	NN	O	O
)	NN	O	O
aa	NN	O	O
;	NN	O	O
relatively	NN	O	O
few	NN	O	O
are	NN	O	O
in	NN	O	O
fully	NN	O	O
conserved	NN	O	O
aa	NN	O	O
(	NN	O	O
where	NN	O	O
they	NN	O	O
might	NN	O	O
be	NN	O	O
lethal	NN	O	O
)	NN	O	O
or	NN	O	O
in	NN	O	O
poorly	NN	O	O
conserved	NN	O	O
aa	NN	O	O
,	NN	O	O
where	NN	O	O
presumably	NN	O	O
they	NN	O	O
simply	NN	O	O
would	NN	O	O
not	NN	O	O
cause	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
all	NN	O	O
human	NN	O	O
mutants	NN	O	O
have	NN	O	O
residual	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
and	NN	O	O
that	NN	O	O
null	NN	O	O
mutations	NN	O	O
are	NN	O	O
lethal	NN	O	O
at	NN	O	O
some	NN	O	O
stage	NN	O	O
of	NN	O	O
development	NN	O	O
.	NN	O	O

Comparing	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	O
housekeeping	NN	O	O
gene	NN	O	O
with	NN	O	O
evolutionary	NN	O	O
conservation	NN	O	O
is	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
pinpointing	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
stability	NN	O	O
or	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
corresponding	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
current	NN	O	O
explosive	NN	O	O
increase	NN	O	O
in	NN	O	O
full	NN	O	O
genome	NN	O	O
sequencing	NN	O	O
projects	NN	O	O
,	NN	O	O
this	NN	O	O
tool	NN	O	O
will	NN	O	O
become	NN	O	O
rapidly	NN	O	O
available	NN	O	O
for	NN	O	O
numerous	NN	O	O
other	NN	O	O
genes	NN	O	O
.	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
and	NN	O	O
regulated	NN	O	O
modes	NN	O	O
of	NN	O	O
splicing	NN	O	O
produce	NN	O	O
six	NN	O	O
major	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
DMPK	NN	O	O
)	NN	O	O
isoforms	NN	O	O
with	NN	O	O
distinct	NN	O	O
properties	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
prevalent	NN	O	O
inherited	NN	O	B-Disease
neuromuscular	NN	O	I-Disease
disease	NN	O	I-Disease
in	NN	O	O
adults	NN	O	O
.	NN	O	O

The	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
CTG	NN	O	O
triplet	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
-	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
DMPK	NN	O	O
)	NN	O	O
gene	NN	O	O
,	NN	O	O
consisting	NN	O	O
of	NN	O	O
15	NN	O	O
exons	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
transgenic	NN	O	O
DMPK	NN	O	O
-	NN	O	O
overexpressor	NN	O	O
mouse	NN	O	O
model	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
endogenous	NN	O	O
mouse	NN	O	O
DMPK	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
human	NN	O	O
DMPK	NN	O	O
transgene	NN	O	O
produce	NN	O	O
six	NN	O	O
major	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
mRNAs	NN	O	O
which	NN	O	O
have	NN	O	O
almost	NN	O	O
identical	NN	O	O
cell	NN	O	O
type	NN	O	O
-	NN	O	O
dependent	NN	O	O
distribution	NN	O	O
frequencies	NN	O	O
and	NN	O	O
expression	NN	O	O
patterns	NN	O	O
.	NN	O	O

Use	NN	O	O
of	NN	O	O
a	NN	O	O
cryptic	NN	O	O
5	NN	O	O
splice	NN	O	O
site	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
absence	NN	O	O
or	NN	O	O
presence	NN	O	O
of	NN	O	O
15	NN	O	O
nucleotides	NN	O	O
specifying	NN	O	O
a	NN	O	O
VSGGG	NN	O	O
peptide	NN	O	O
motif	NN	O	O
,	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
cryptic	NN	O	O
3	NN	O	O
splice	NN	O	O
site	NN	O	O
in	NN	O	O
exon	NN	O	O
14	NN	O	O
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
frameshift	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
reading	NN	O	O
frame	NN	O	O
,	NN	O	O
occur	NN	O	O
as	NN	O	O
independent	NN	O	O
stochastic	NN	O	O
events	NN	O	O
in	NN	O	O
all	NN	O	O
tissues	NN	O	O
examined	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
excision	NN	O	O
of	NN	O	O
exons	NN	O	O
13	NN	O	O
/	NN	O	O
14	NN	O	O
that	NN	O	O
causes	NN	O	O
a	NN	O	O
frameshift	NN	O	O
and	NN	O	O
creates	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
terminally	NN	O	O
truncated	NN	O	O
protein	NN	O	O
is	NN	O	O
clearly	NN	O	O
cell	NN	O	O
type	NN	O	O
dependent	NN	O	O
and	NN	O	O
occurs	NN	O	O
predominantly	NN	O	O
in	NN	O	O
smooth	NN	O	O
muscle	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
all	NN	O	O
six	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
mouse	NN	O	O
cDNAs	NN	O	O
that	NN	O	O
result	NN	O	O
from	NN	O	O
combinations	NN	O	O
of	NN	O	O
these	NN	O	O
three	NN	O	O
major	NN	O	O
splicing	NN	O	O
events	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
their	NN	O	O
transfection	NN	O	O
into	NN	O	O
cells	NN	O	O
in	NN	O	O
culture	NN	O	O
leads	NN	O	O
to	NN	O	O
production	NN	O	O
of	NN	O	O
four	NN	O	O
different	NN	O	O
approximately	NN	O	O
74	NN	O	O
kDa	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
(	NN	O	O
heart	NN	O	O
-	NN	O	O
,	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
-	NN	O	O
or	NN	O	O
brain	NN	O	O
-	NN	O	O
specific	NN	O	O
)	NN	O	O
and	NN	O	O
two	NN	O	O
C	NN	O	O
-	NN	O	O
terminally	NN	O	O
truncated	NN	O	O
approximately	NN	O	O
68	NN	O	O
kDa	NN	O	O
(	NN	O	O
smooth	NN	O	O
muscle	NN	O	O
-	NN	O	O
specific	NN	O	O
)	NN	O	O
isoforms	NN	O	O
.	NN	O	O

Information	NN	O	O
on	NN	O	O
DMPK	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
isoform	NN	O	O
expression	NN	O	O
patterns	NN	O	O
will	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
recognizing	NN	O	O
differential	NN	O	O
effects	NN	O	O
of	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
(	NN	O	O
n	NN	O	O
)	NN	O	O
expansion	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
manifestation	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
,	NN	O	O
phenotypic	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
and	NN	O	O
risk	NN	O	O
of	NN	O	O
venous	NN	O	B-Disease
thrombosis	NN	O	I-Disease
in	NN	O	O
families	NN	O	O
with	NN	O	O
inherited	NN	O	O
deficiencies	NN	O	B-Disease
of	NN	O	I-Disease
protein	NN	O	I-Disease
S	NN	O	I-Disease
.	NN	O	O

Protein	NN	O	B-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
recognized	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
venous	NN	O	B-Disease
thrombosis	NN	O	I-Disease
.	NN	O	O

Of	NN	O	O
all	NN	O	O
the	NN	O	O
inherited	NN	O	O
thrombophilic	NN	O	B-Disease
conditions	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
remains	NN	O	O
the	NN	O	O
most	NN	O	O
difficult	NN	O	O
to	NN	O	O
diagnose	NN	O	O
because	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
variability	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
inconclusive	NN	O	O
results	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
overcome	NN	O	O
this	NN	O	O
problem	NN	O	O
by	NN	O	O
studying	NN	O	O
a	NN	O	O
cohort	NN	O	O
of	NN	O	O
patients	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
center	NN	O	O
where	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
was	NN	O	O
confirmed	NN	O	O
at	NN	O	O
the	NN	O	O
genetic	NN	O	O
level	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
eight	NN	O	O
index	NN	O	O
patients	NN	O	O
with	NN	O	O
protein	NN	O	B-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
PROS1	NN	O	B-Disease
gene	NN	O	I-Disease
defect	NN	O	I-Disease
were	NN	O	O
studied	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
109	NN	O	O
first	NN	O	O
-	NN	O	O
degree	NN	O	O
relatives	NN	O	O
.	NN	O	O

To	NN	O	O
avoid	NN	O	O
selection	NN	O	O
bias	NN	O	O
,	NN	O	O
we	NN	O	O
confined	NN	O	O
analysis	NN	O	O
of	NN	O	O
total	NN	O	O
and	NN	O	O
free	NN	O	O
protein	NN	O	O
S	NN	O	O
levels	NN	O	O
and	NN	O	O
thrombotic	NN	O	O
risk	NN	O	O
to	NN	O	O
the	NN	O	O
patients	NN	O	O
relatives	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
group	NN	O	O
of	NN	O	O
relatives	NN	O	O
,	NN	O	O
a	NN	O	O
low	NN	O	O
free	NN	O	O
protein	NN	O	O
S	NN	O	O
level	NN	O	O
was	NN	O	O
the	NN	O	O
most	NN	O	O
reliable	NN	O	O
predictor	NN	O	O
of	NN	O	O
a	NN	O	O
PROS1	NN	O	B-Disease
gene	NN	O	I-Disease
defect	NN	O	I-Disease
(	NN	O	O
sensitivity	NN	O	O
97	NN	O	O
.	NN	O	O
7	NN	O	O
%	NN	O	O
,	NN	O	O
specificity	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

First	NN	O	O
-	NN	O	O
degree	NN	O	O
relatives	NN	O	O
with	NN	O	O
a	NN	O	O
PROS1	NN	O	B-Disease
gene	NN	O	I-Disease
defect	NN	O	I-Disease
had	NN	O	O
a	NN	O	O
5	NN	O	O
.	NN	O	O

0	NN	O	O
-	NN	O	O
fold	NN	O	O
higher	NN	O	O
risk	NN	O	O
of	NN	O	O
thrombosis	NN	O	B-Disease
(	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
,	NN	O	O
1	NN	O	O
.	NN	O	O
5	NN	O	O
-	NN	O	O
16	NN	O	O
.	NN	O	O
8	NN	O	O
)	NN	O	O
than	NN	O	O
those	NN	O	O
with	NN	O	O
a	NN	O	O
normal	NN	O	O
PROS1	NN	O	O
gene	NN	O	O
and	NN	O	O
no	NN	O	O
other	NN	O	O
recognized	NN	O	O
thrombophilic	NN	O	B-Disease
defect	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
pregnancy	NN	O	O
/	NN	O	O
puerperium	NN	O	O
and	NN	O	O
immobility	NN	O	O
/	NN	O	O
trauma	NN	O	B-Disease
were	NN	O	O
important	NN	O	O
precipitating	NN	O	O
factors	NN	O	O
for	NN	O	O
thrombosis	NN	O	B-Disease
,	NN	O	O
almost	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
events	NN	O	O
were	NN	O	O
spontaneous	NN	O	O
.	NN	O	O

Relatives	NN	O	O
with	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
or	NN	O	O
major	NN	O	O
structural	NN	O	O
defects	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
PROS1	NN	O	I-Disease
gene	NN	O	I-Disease
were	NN	O	O
more	NN	O	O
likely	NN	O	O
to	NN	O	O
have	NN	O	O
had	NN	O	O
a	NN	O	O
thrombotic	NN	O	O
event	NN	O	O
and	NN	O	O
had	NN	O	O
significantly	NN	O	O
lower	NN	O	O
total	NN	O	O
and	NN	O	O
free	NN	O	O
protein	NN	O	O
S	NN	O	O
levels	NN	O	O
than	NN	O	O
those	NN	O	O
relatives	NN	O	O
having	NN	O	O
missense	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
persons	NN	O	O
with	NN	O	O
PROS1	NN	O	B-Disease
gene	NN	O	I-Disease
defects	NN	O	I-Disease
and	NN	O	O
protein	NN	O	B-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
are	NN	O	O
at	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
thrombosis	NN	O	B-Disease
and	NN	O	O
that	NN	O	O
free	NN	O	O
protein	NN	O	O
S	NN	O	O
estimation	NN	O	O
offers	NN	O	O
the	NN	O	O
most	NN	O	O
reliable	NN	O	O
way	NN	O	O
of	NN	O	O
diagnosing	NN	O	O
the	NN	O	O
deficiency	NN	O	O
.	NN	O	O

(	NN	O	O
Blood	NN	O	O
.	NN	O	O
2000	NN	O	O
;	NN	O	O
95	NN	O	O
1935	NN	O	O
-	NN	O	O
1941	NN	O	O
)	NN	O	O
.	NN	O	O

Autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ALPS	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
a	NN	O	O
child	NN	O	O
from	NN	O	O
consanguineous	NN	O	O
parents	NN	O	O
:	NN	O	O
a	NN	O	O
dominant	NN	O	O
or	NN	O	O
recessive	NN	O	O
disease	NN	O	O
?	NN	O	O

Autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ALPS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
autoimmune	NN	O	O
features	NN	O	O
and	NN	O	O
lymphoproliferations	NN	O	O
and	NN	O	O
is	NN	O	O
generally	NN	O	O
caused	NN	O	O
by	NN	O	O
defective	NN	O	O
Fas	NN	O	O
-	NN	O	O
mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
describes	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
ALPS	NN	O	B-Disease
without	NN	O	O
detectable	NN	O	O
Fas	NN	O	O
expression	NN	O	O
on	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
FAS	NN	O	O
transcripts	NN	O	O
via	NN	O	O
real	NN	O	O
-	NN	O	O
time	NN	O	O
quantitative	NN	O	O
PCR	NN	O	O
made	NN	O	O
a	NN	O	O
severe	NN	O	O
transcriptional	NN	O	O
defect	NN	O	O
unlikely	NN	O	O
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
FAS	NN	O	O
gene	NN	O	O
revealed	NN	O	O
a	NN	O	O
20	NN	O	O
-	NN	O	O
nucleotide	NN	O	O
duplication	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
exon	NN	O	O
affecting	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
signaling	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
homozygous	NN	O	O
for	NN	O	O
this	NN	O	O
mutation	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
consanguineous	NN	O	O
parents	NN	O	O
and	NN	O	O
the	NN	O	O
siblings	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
reported	NN	O	O
here	NN	O	O
is	NN	O	O
a	NN	O	O
human	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
Fas	NN	O	O
-	NN	O	O
null	NN	O	O
mouse	NN	O	O
,	NN	O	O
inasmuch	NN	O	O
as	NN	O	O
she	NN	O	O
carries	NN	O	O
an	NN	O	O
autosomal	NN	O	O
homozygous	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
FAS	NN	O	O
gene	NN	O	O
and	NN	O	O
she	NN	O	O
shows	NN	O	O
the	NN	O	O
severe	NN	O	O
and	NN	O	O
accelerated	NN	O	O
ALPS	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
heterozygous	NN	O	O
family	NN	O	O
members	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
the	NN	O	O
ALPS	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
FAS	NN	O	O
mutation	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
is	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
novel	NN	O	O
imprinted	NN	O	O
transcripts	NN	O	O
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
deletion	NN	O	O
region	NN	O	O
:	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
regional	NN	O	O
imprinting	NN	O	O
control	NN	O	O
.	NN	O	O

Deletions	NN	O	O
and	NN	O	O
other	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
two	NN	O	O
developmental	NN	O	O
disorders	NN	O	O
,	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
imprinted	NN	O	O
,	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
genes	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
imprinted	NN	O	O
genes	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
PWS	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
the	NN	O	O
range	NN	O	O
over	NN	O	O
which	NN	O	O
the	NN	O	O
imprinting	NN	O	O
signal	NN	O	O
acts	NN	O	O
to	NN	O	O
silence	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
parent	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
origin	NN	O	O
-	NN	O	O
specific	NN	O	O
manner	NN	O	O
,	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
additional	NN	O	O
imprinted	NN	O	O
genes	NN	O	O
that	NN	O	O
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
phenotype	NN	O	O
and	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
regional	NN	O	O
control	NN	O	O
of	NN	O	O
imprinting	NN	O	O
in	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
constructed	NN	O	O
an	NN	O	O
imprinted	NN	O	O
transcript	NN	O	O
map	NN	O	O
of	NN	O	O
the	NN	O	O
PWS	NN	O	O
-	NN	O	O
AS	NN	O	O
deletion	NN	O	O
interval	NN	O	O
.	NN	O	O

The	NN	O	O
imprinting	NN	O	O
status	NN	O	O
of	NN	O	O
22	NN	O	O
expressed	NN	O	O
sequence	NN	O	O
tags	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
radiation	NN	O	O
-	NN	O	O
hybrid	NN	O	O
human	NN	O	O
transcript	NN	O	O
maps	NN	O	O
or	NN	O	O
physical	NN	O	O
maps	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
a	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
-	NN	O	O
PCR	NN	O	O
assay	NN	O	O
and	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
transcripts	NN	O	O
on	NN	O	O
the	NN	O	O
physical	NN	O	O
map	NN	O	O
.	NN	O	O

Seven	NN	O	O
new	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
transcripts	NN	O	O
localize	NN	O	O
to	NN	O	O
an	NN	O	O
approximately	NN	O	O
1	NN	O	O
.	NN	O	O

5	NN	O	O
-	NN	O	O
Mb	NN	O	O
domain	NN	O	O
surrounding	NN	O	O
the	NN	O	O
SNRPN	NN	O	O
-	NN	O	O
associated	NN	O	O
imprinting	NN	O	O
center	NN	O	O
,	NN	O	O
which	NN	O	O
already	NN	O	O
includes	NN	O	O
four	NN	O	O
imprinted	NN	O	O
,	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
genes	NN	O	O
.	NN	O	O

All	NN	O	O
other	NN	O	O
tested	NN	O	O
new	NN	O	O
transcripts	NN	O	O
in	NN	O	O
the	NN	O	O
deletion	NN	O	O
region	NN	O	O
were	NN	O	O
expressed	NN	O	O
from	NN	O	O
both	NN	O	O
alleles	NN	O	O
.	NN	O	O

A	NN	O	O
domain	NN	O	O
of	NN	O	O
exclusive	NN	O	O
paternal	NN	O	O
expression	NN	O	O
surrounding	NN	O	O
the	NN	O	O
imprinting	NN	O	O
center	NN	O	O
suggests	NN	O	O
strong	NN	O	O
regional	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
imprinting	NN	O	O
process	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
provides	NN	O	O
the	NN	O	O
means	NN	O	O
for	NN	O	O
further	NN	O	O
investigation	NN	O	O
of	NN	O	O
additional	NN	O	O
genes	NN	O	O
that	NN	O	O
cause	NN	O	O
or	NN	O	O
modify	NN	O	O
the	NN	O	O
phenotypes	NN	O	O
associated	NN	O	O
with	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
.	NN	O	O

Combined	NN	O	O
analysis	NN	O	O
of	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
linkage	NN	O	O
to	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
:	NN	O	O
results	NN	O	O
from	NN	O	O
772	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
from	NN	O	O
the	NN	O	O
International	NN	O	O
Consortium	NN	O	O
for	NN	O	O
Prostate	NN	O	B-Disease
Cancer	NN	O	I-Disease
Genetics	NN	O	O
.	NN	O	O

A	NN	O	O
previous	NN	O	O
linkage	NN	O	O
study	NN	O	O
provided	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
-	NN	O	O
susceptibility	NN	O	O
locus	NN	O	O
at	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
.	NN	O	O

Subsequent	NN	O	O
reports	NN	O	O
in	NN	O	O
additional	NN	O	O
collections	NN	O	O
of	NN	O	O
families	NN	O	O
have	NN	O	O
yielded	NN	O	O
conflicting	NN	O	O
results	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
evidence	NN	O	O
for	NN	O	O
locus	NN	O	O
heterogeneity	NN	O	O
has	NN	O	O
been	NN	O	O
provided	NN	O	O
by	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
other	NN	O	O
putative	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
loci	NN	O	O
on	NN	O	O
Xq27	NN	O	O
-	NN	O	O
28	NN	O	O
,	NN	O	O
1q42	NN	O	O
-	NN	O	O
43	NN	O	O
,	NN	O	O
and	NN	O	O
1p36	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
describes	NN	O	O
a	NN	O	O
combined	NN	O	O
analysis	NN	O	O
for	NN	O	O
six	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
region	NN	O	O
in	NN	O	O
772	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
ascertained	NN	O	O
by	NN	O	O
the	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
International	NN	O	O
Consortium	NN	O	O
for	NN	O	O
Prostate	NN	O	B-Disease
Cancer	NN	O	I-Disease
Genetics	NN	O	O
(	NN	O	O
ICPCG	NN	O	O
)	NN	O	O
from	NN	O	O
North	NN	O	O
America	NN	O	O
,	NN	O	O
Australia	NN	O	O
,	NN	O	O
Finland	NN	O	O
,	NN	O	O
Norway	NN	O	O
,	NN	O	O
Sweden	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
United	NN	O	O
Kingdom	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
some	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
peak	NN	O	O
parametric	NN	O	O
multipoint	NN	O	O
LOD	NN	O	O
score	NN	O	O
assuming	NN	O	O
heterogeneity	NN	O	O
(	NN	O	O
HLOD	NN	O	O
)	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

40	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
01	NN	O	O
)	NN	O	O
at	NN	O	O
D1S212	NN	O	O
.	NN	O	O

The	NN	O	O
estimated	NN	O	O
proportion	NN	O	O
of	NN	O	O
families	NN	O	O
(	NN	O	O
alpha	NN	O	O
)	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
locus	NN	O	O
was	NN	O	O
.	NN	O	O

06	NN	O	O
(	NN	O	O
1	NN	O	O
-	NN	O	O
LOD	NN	O	O
support	NN	O	O
interval	NN	O	O
.	NN	O	O
01	NN	O	O
-	NN	O	O
.	NN	O	O
12	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
evidence	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
by	NN	O	O
a	NN	O	O
nonparametric	NN	O	O
approach	NN	O	O
,	NN	O	O
presumably	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
extensive	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

Further	NN	O	O
parametric	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
male	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
male	NN	O	O
disease	NN	O	O
transmission	NN	O	O
within	NN	O	O
the	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
subset	NN	O	O
of	NN	O	O
491	NN	O	O
such	NN	O	O
families	NN	O	O
,	NN	O	O
the	NN	O	O
peak	NN	O	O
HLOD	NN	O	O
was	NN	O	O
2	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
subset	NN	O	O
of	NN	O	O
491	NN	O	O
such	NN	O	O
families	NN	O	O
,	NN	O	O
the	NN	O	O
peak	NN	O	O
HLOD	NN	O	O
was	NN	O	O
2	NN	O	O
.	NN	O	O

56	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
0006	NN	O	O
)	NN	O	O
and	NN	O	O
alpha	NN	O	O
=	NN	O	O
.	NN	O	O

11	NN	O	O
(	NN	O	O
1	NN	O	O
-	NN	O	O
LOD	NN	O	O
support	NN	O	O
interval	NN	O	O
.	NN	O	O
04	NN	O	O
-	NN	O	O
.	NN	O	O
19	NN	O	O
)	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
HLODs	NN	O	O
of	NN	O	O
0	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
281	NN	O	O
families	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
families	NN	O	O
with	NN	O	O
male	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
male	NN	O	O
disease	NN	O	O
transmission	NN	O	O
,	NN	O	O
alpha	NN	O	O
increased	NN	O	O
with	NN	O	O
the	NN	O	O
early	NN	O	O
mean	NN	O	O
age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
(	NN	O	O
<	NN	O	O
65	NN	O	O
years	NN	O	O
,	NN	O	O
alpha	NN	O	O
=	NN	O	O
.	NN	O	O
19	NN	O	O
,	NN	O	O
with	NN	O	O
1	NN	O	O
-	NN	O	O
LOD	NN	O	O
support	NN	O	O
interval	NN	O	O
.	NN	O	O
06	NN	O	O
-	NN	O	O
.	NN	O	O
34	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
affected	NN	O	O
family	NN	O	O
members	NN	O	O
(	NN	O	O
five	NN	O	O
or	NN	O	O
more	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
alpha	NN	O	O
=	NN	O	O
.	NN	O	O
15	NN	O	O
,	NN	O	O
with	NN	O	O
1	NN	O	O
-	NN	O	O
LOD	NN	O	O
support	NN	O	O
interval	NN	O	O
.	NN	O	O
04	NN	O	O
-	NN	O	O
.	NN	O	O
28	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
highest	NN	O	O
value	NN	O	O
of	NN	O	O
alpha	NN	O	O
was	NN	O	O
observed	NN	O	O
for	NN	O	O
the	NN	O	O
48	NN	O	O
families	NN	O	O
that	NN	O	O
met	NN	O	O
all	NN	O	O
three	NN	O	O
criteria	NN	O	O
(	NN	O	O
peak	NN	O	O
HLOD	NN	O	O
=	NN	O	O
2	NN	O	O
.	NN	O	O
25	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
001	NN	O	O
,	NN	O	O
alpha	NN	O	O
=	NN	O	O
.	NN	O	O
29	NN	O	O
,	NN	O	O
with	NN	O	O
1	NN	O	O
-	NN	O	O
LOD	NN	O	O
support	NN	O	O
interval	NN	O	O
.	NN	O	O
08	NN	O	O
-	NN	O	O
.	NN	O	O
53	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
finding	NN	O	O
of	NN	O	O
a	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
-	NN	O	O
susceptibility	NN	O	O
gene	NN	O	O
linked	NN	O	O
to	NN	O	O
1q24	NN	O	O
-	NN	O	O
25	NN	O	O
,	NN	O	O
albeit	NN	O	O
in	NN	O	O
a	NN	O	O
defined	NN	O	O
subset	NN	O	O
of	NN	O	O
prostate	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

Although	NN	O	O
HPC1	NN	O	O
accounts	NN	O	O
for	NN	O	O
only	NN	O	O
a	NN	O	O
small	NN	O	O
proportion	NN	O	O
of	NN	O	O
all	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
hereditary	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
more	NN	O	O
prominent	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
subset	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
several	NN	O	O
members	NN	O	O
affected	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
age	NN	O	O
and	NN	O	O
with	NN	O	O
male	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
male	NN	O	O
disease	NN	O	O
transmission	NN	O	O
.	NN	O	O

A	NN	O	O
recurrent	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
maternal	NN	O	O
allele	NN	O	O
with	NN	O	O
36	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
causes	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
two	NN	O	O
sisters	NN	O	O
.	NN	O	O

Large	NN	O	O
intergenerational	NN	O	O
repeat	NN	O	O
expansions	NN	O	O
of	NN	O	O
the	NN	O	O
CAG	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
have	NN	O	O
been	NN	O	O
well	NN	O	O
documented	NN	O	O
for	NN	O	O
the	NN	O	O
male	NN	O	O
germline	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
recurrent	NN	O	O
large	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
maternal	NN	O	O
allele	NN	O	O
with	NN	O	O
36	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
(	NN	O	O
to	NN	O	O
66	NN	O	O
and	NN	O	O
57	NN	O	O
repeats	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
in	NN	O	O
two	NN	O	O
daughters	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
onset	NN	O	O
of	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
and	NN	O	O
third	NN	O	O
decade	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
without	NN	O	O
history	NN	O	O
of	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
give	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
gonadal	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
the	NN	O	O
unaffected	NN	O	O
mother	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
large	NN	O	O
expansions	NN	O	O
also	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
female	NN	O	O
germline	NN	O	O
and	NN	O	O
that	NN	O	O
a	NN	O	O
negative	NN	O	O
selection	NN	O	O
of	NN	O	O
oocytes	NN	O	O
with	NN	O	O
long	NN	O	O
repeats	NN	O	O
might	NN	O	O
explain	NN	O	O
the	NN	O	O
different	NN	O	O
instability	NN	O	O
behavior	NN	O	O
of	NN	O	O
the	NN	O	O
male	NN	O	O
and	NN	O	O
the	NN	O	O
female	NN	O	O
germlines	NN	O	O
.	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
development	NN	O	O
of	NN	O	O
Purkinje	NN	O	O
cells	NN	O	O
and	NN	O	O
lymphocytes	NN	O	O
in	NN	O	O
Atm	NN	O	O
mutant	NN	O	O
mice	NN	O	O
.	NN	O	O

Motor	NN	O	B-Disease
incoordination	NN	O	I-Disease
,	NN	O	O
immune	NN	O	B-Disease
deficiencies	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
are	NN	O	O
the	NN	O	O
characteristic	NN	O	O
features	NN	O	O
of	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
disease	NN	O	I-Disease
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
.	NN	O	O

Through	NN	O	O
gene	NN	O	O
targeting	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
line	NN	O	O
of	NN	O	O
Atm	NN	O	O
mutant	NN	O	O
mice	NN	O	O
,	NN	O	O
Atm	NN	O	O
(	NN	O	O
y	NN	O	O
/	NN	O	O
y	NN	O	O
)	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
other	NN	O	O
Atm	NN	O	O
mutant	NN	O	O
mice	NN	O	O
,	NN	O	O
Atm	NN	O	O
(	NN	O	O
y	NN	O	O
/	NN	O	O
y	NN	O	O
)	NN	O	O
mice	NN	O	O
show	NN	O	O
a	NN	O	O
lower	NN	O	O
incidence	NN	O	O
of	NN	O	O
thymic	NN	O	B-Disease
lymphoma	NN	O	I-Disease
and	NN	O	O
survive	NN	O	O
beyond	NN	O	O
a	NN	O	O
few	NN	O	O
months	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

Atm	NN	O	O
(	NN	O	O
y	NN	O	O
/	NN	O	O
y	NN	O	O
)	NN	O	O
mice	NN	O	O
exhibit	NN	O	O
deficits	NN	O	O
in	NN	O	O
motor	NN	O	O
learning	NN	O	O
indicative	NN	O	O
of	NN	O	O
cerebellar	NN	O	B-Disease
dysfunction	NN	O	I-Disease
.	NN	O	O

Even	NN	O	O
though	NN	O	O
we	NN	O	O
found	NN	O	O
no	NN	O	O
gross	NN	O	O
cerebellar	NN	O	B-Disease
degeneration	NN	O	I-Disease
in	NN	O	O
older	NN	O	O
Atm	NN	O	O
(	NN	O	O
y	NN	O	O
/	NN	O	O
y	NN	O	O
)	NN	O	O
animals	NN	O	O
,	NN	O	O
ectopic	NN	O	O
and	NN	O	O
abnormally	NN	O	O
differentiated	NN	O	O
Purkinje	NN	O	O
cells	NN	O	O
were	NN	O	O
apparent	NN	O	O
in	NN	O	O
mutant	NN	O	O
mice	NN	O	O
of	NN	O	O
all	NN	O	O
ages	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
establish	NN	O	O
that	NN	O	O
some	NN	O	O
neuropathological	NN	O	B-Disease
abnormalities	NN	O	I-Disease
seen	NN	O	O
in	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
also	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
Atm	NN	O	O
mutant	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
effect	NN	O	O
of	NN	O	O
Atm	NN	O	B-Disease
deficiency	NN	O	I-Disease
on	NN	O	O
development	NN	O	O
or	NN	O	O
maintenance	NN	O	O
of	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
thymocytes	NN	O	O
.	NN	O	O

We	NN	O	O
discuss	NN	O	O
these	NN	O	O
findings	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
abnormal	NN	O	O
development	NN	O	O
of	NN	O	O
Purkinje	NN	O	O
cells	NN	O	O
and	NN	O	O
lymphocytes	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Novel	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
ATP7B	NN	O	O
gene	NN	O	O
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
WD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
copper	NN	O	O
accumulation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
,	NN	O	O
brain	NN	O	O
,	NN	O	O
kidneys	NN	O	O
,	NN	O	O
and	NN	O	O
corneas	NN	O	O
,	NN	O	O
and	NN	O	O
culminating	NN	O	O
in	NN	O	O
copper	NN	O	O
toxication	NN	O	O
in	NN	O	O
these	NN	O	O
organs	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
responsible	NN	O	O
gene	NN	O	O
,	NN	O	O
ATP7B	NN	O	O
,	NN	O	O
in	NN	O	O
four	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
WD	NN	O	B-Disease
.	NN	O	O

By	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
five	NN	O	O
mutations	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
two	NN	O	O
were	NN	O	O
novel	NN	O	O
,	NN	O	O
and	NN	O	O
16	NN	O	O
polymorphisms	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
6	NN	O	O
were	NN	O	O
novel	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
2871delC	NN	O	O
and	NN	O	O
2513delA	NN	O	O
shift	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
so	NN	O	O
that	NN	O	O
truncated	NN	O	O
abnormal	NN	O	O
protein	NN	O	O
is	NN	O	O
expected	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
these	NN	O	O
mutations	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
hepatic	NN	O	O
-	NN	O	O
type	NN	O	O
of	NN	O	O
early	NN	O	O
onset	NN	O	O
,	NN	O	O
the	NN	O	O
mutations	NN	O	O
A874V	NN	O	O
,	NN	O	O
R778L	NN	O	O
,	NN	O	O
and	NN	O	O
3892delGTC	NN	O	O
were	NN	O	O
either	NN	O	O
missense	NN	O	O
mutations	NN	O	O
or	NN	O	O
in	NN	O	O
frame	NN	O	O
1	NN	O	O
-	NN	O	O
amino	NN	O	O
acid	NN	O	O
deletion	NN	O	O
,	NN	O	O
and	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
hepato	NN	O	O
-	NN	O	O
neurologic	NN	O	O
type	NN	O	O
of	NN	O	O
late	NN	O	O
onset	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
2871delC	NN	O	O
and	NN	O	O
R778L	NN	O	O
have	NN	O	O
been	NN	O	O
previously	NN	O	O
reported	NN	O	O
in	NN	O	O
a	NN	O	O
relatively	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
R778L	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
prevalent	NN	O	O
in	NN	O	O
Asian	NN	O	O
countries	NN	O	O
than	NN	O	O
in	NN	O	O
other	NN	O	O
countries	NN	O	O
of	NN	O	O
the	NN	O	O
world	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
mutations	NN	O	O
tend	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
types	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
WD	NN	O	B-Disease
will	NN	O	O
facilitate	NN	O	O
the	NN	O	O
fast	NN	O	O
and	NN	O	O
effective	NN	O	O
genetic	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
WD	NN	O	B-Disease
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Autoinhibition	NN	O	O
and	NN	O	O
activation	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
protein	NN	O	O
.	NN	O	O

The	NN	O	O
Rho	NN	O	O
-	NN	O	O
family	NN	O	O
GTPase	NN	O	O
,	NN	O	O
Cdc42	NN	O	O
,	NN	O	O
can	NN	O	O
regulate	NN	O	O
the	NN	O	O
actin	NN	O	O
cytoskeleton	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
protein	NN	O	O
(	NN	O	O
WASP	NN	O	O
)	NN	O	O
family	NN	O	O
members	NN	O	O
.	NN	O	O

Activation	NN	O	O
relieves	NN	O	O
an	NN	O	O
autoinhibitory	NN	O	O
contact	NN	O	O
between	NN	O	O
the	NN	O	O
GTPase	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
and	NN	O	O
the	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminal	NN	O	O
region	NN	O	O
of	NN	O	O
WASP	NN	O	O
proteins	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
autoinhibited	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
GTPase	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
WASP	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
region	NN	O	O
or	NN	O	O
by	NN	O	O
organic	NN	O	O
co	NN	O	O
-	NN	O	O
solvents	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
autoinhibited	NN	O	O
complex	NN	O	O
,	NN	O	O
intramolecular	NN	O	O
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
GTPase	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
occlude	NN	O	O
residues	NN	O	O
of	NN	O	O
the	NN	O	O
C	NN	O	O
terminus	NN	O	O
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
Arp2	NN	O	O
/	NN	O	O
3	NN	O	O
actin	NN	O	O
-	NN	O	O
nucleating	NN	O	O
complex	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
Cdc42	NN	O	O
to	NN	O	O
the	NN	O	O
GTPase	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
causes	NN	O	O
a	NN	O	O
dramatic	NN	O	O
conformational	NN	O	O
change	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
hydrophobic	NN	O	O
core	NN	O	O
and	NN	O	O
release	NN	O	O
of	NN	O	O
the	NN	O	O
C	NN	O	O
terminus	NN	O	O
,	NN	O	O
enabling	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
actin	NN	O	O
regulatory	NN	O	O
machinery	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
intrinsically	NN	O	O
unstructured	NN	O	O
peptides	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
GTPase	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
WASP	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
into	NN	O	O
distinct	NN	O	O
structural	NN	O	O
and	NN	O	O
functional	NN	O	O
states	NN	O	O
depending	NN	O	O
on	NN	O	O
context	NN	O	O
.	NN	O	O
.	NN	O	O

hCds1	NN	O	O
-	NN	O	O
mediated	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
regulates	NN	O	O
the	NN	O	O
DNA	NN	O	O
damage	NN	O	O
response	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
with	NN	O	O
inherited	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
and	NN	O	O
about	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
with	NN	O	O
only	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
the	NN	O	O
biochemical	NN	O	O
function	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
is	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
DNA	NN	O	O
damage	NN	O	O
repair	NN	O	O
and	NN	O	O
cell	NN	O	O
-	NN	O	O
cycle	NN	O	O
checkpoint	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
exists	NN	O	O
in	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
but	NN	O	O
is	NN	O	O
hyperphosphorylated	NN	O	O
and	NN	O	O
disperses	NN	O	O
after	NN	O	O
DNA	NN	O	O
damage	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
whether	NN	O	O
BRCA1	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
dispersion	NN	O	O
and	NN	O	O
its	NN	O	O
function	NN	O	O
in	NN	O	O
DNA	NN	O	O
damage	NN	O	O
response	NN	O	O
are	NN	O	O
related	NN	O	O
.	NN	O	O

In	NN	O	O
yeast	NN	O	O
the	NN	O	O
DNA	NN	O	O
damage	NN	O	O
response	NN	O	O
and	NN	O	O
the	NN	O	O
replication	NN	O	O
-	NN	O	O
block	NN	O	O
checkpoint	NN	O	O
are	NN	O	O
mediated	NN	O	O
partly	NN	O	O
through	NN	O	O
the	NN	O	O
Cds1	NN	O	O
kinase	NN	O	O
family	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	O
Cds1	NN	O	O
kinase	NN	O	O
(	NN	O	O
hCds1	NN	O	O
/	NN	O	O
Chk2	NN	O	O
)	NN	O	O
regulates	NN	O	O
BRCA1	NN	O	O
function	NN	O	O
after	NN	O	O
DNA	NN	O	O
damage	NN	O	O
by	NN	O	O
phosphorylating	NN	O	O
serine	NN	O	O
988	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
hCds1	NN	O	O
and	NN	O	O
BRCA1	NN	O	O
interact	NN	O	O
and	NN	O	O
co	NN	O	O
-	NN	O	O
localize	NN	O	O
within	NN	O	O
discrete	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
but	NN	O	O
separate	NN	O	O
after	NN	O	O
gamma	NN	O	O
irradiation	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
at	NN	O	O
serine	NN	O	O
988	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
from	NN	O	O
hCds1	NN	O	O
.	NN	O	O

This	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
also	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
to	NN	O	O
restore	NN	O	O
survival	NN	O	O
after	NN	O	O
DNA	NN	O	O
damage	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
mutated	NN	O	O
cell	NN	O	O
line	NN	O	O
HCC1937	NN	O	O
.	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
rat	NN	O	O
spinocerebellar	NN	O	O
ataxia	NN	O	O
type	NN	O	O
3	NN	O	O
gene	NN	O	O
.	NN	O	O

Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
belongs	NN	O	O
to	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
clinically	NN	O	O
and	NN	O	O
genetically	NN	O	O
heterogeneous	NN	O	O
neurodegenerative	NN	O	B-Disease
disorders	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
progressive	NN	O	B-Disease
cerebellar	NN	O	I-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
mutation	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
unstable	NN	O	O
and	NN	O	O
expanded	NN	O	O
(	NN	O	O
CAG	NN	O	O
)	NN	O	O
n	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

In	NN	O	O
Caucasians	NN	O	O
,	NN	O	O
repeat	NN	O	O
expansions	NN	O	O
in	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
have	NN	O	O
also	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
spinocerebellar	NN	O	B-Disease
ataxia	NN	O	I-Disease
type	NN	O	I-Disease
3	NN	O	I-Disease
(	NN	O	O
SCA3	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
gain	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
the	NN	O	O
MJD1	NN	O	O
/	NN	O	O
SCA3	NN	O	O
gene	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
rat	NN	O	O
homologue	NN	O	O
and	NN	O	O
studied	NN	O	O
its	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
rat	NN	O	O
and	NN	O	O
human	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
genes	NN	O	O
are	NN	O	O
highly	NN	O	O
homologous	NN	O	O
with	NN	O	O
an	NN	O	O
overall	NN	O	O
sequence	NN	O	O
identity	NN	O	O
of	NN	O	O
approximately	NN	O	O
88	NN	O	O
%	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
protein	NN	O	O
differs	NN	O	O
strongly	NN	O	O
from	NN	O	O
the	NN	O	O
published	NN	O	O
human	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
(	NN	O	O
CAG	NN	O	O
)	NN	O	O
n	NN	O	O
block	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
cDNA	NN	O	O
consists	NN	O	O
of	NN	O	O
just	NN	O	O
three	NN	O	O
interrupted	NN	O	O
units	NN	O	O
suggesting	NN	O	O
that	NN	O	O
a	NN	O	O
long	NN	O	O
polyglutamine	NN	O	O
stretch	NN	O	O
is	NN	O	O
not	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
normal	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
ataxin	NN	O	O
-	NN	O	O
3	NN	O	O
protein	NN	O	O
in	NN	O	O
rodents	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
the	NN	O	O
SCA3	NN	O	O
gene	NN	O	O
in	NN	O	O
various	NN	O	O
rat	NN	O	O
and	NN	O	O
human	NN	O	O
tissues	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
.	NN	O	O

The	NN	O	O
mature	NN	O	O
transcript	NN	O	O
is	NN	O	O
approximately	NN	O	O
6	NN	O	O
kb	NN	O	O
in	NN	O	O
length	NN	O	O
.	NN	O	O

In	NN	O	O
rat	NN	O	O
testis	NN	O	O
,	NN	O	O
a	NN	O	O
smaller	NN	O	O
transcript	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

3	NN	O	O
kb	NN	O	O
was	NN	O	O
identified	NN	O	O
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
rsca3	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
most	NN	O	O
rat	NN	O	O
tissues	NN	O	O
including	NN	O	O
brain	NN	O	O
.	NN	O	O

Analyzing	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
SCA3	NN	O	O
gene	NN	O	O
in	NN	O	O
several	NN	O	O
human	NN	O	O
brain	NN	O	O
sections	NN	O	O
revealed	NN	O	O
no	NN	O	O
significant	NN	O	O
higher	NN	O	O
mRNA	NN	O	O
level	NN	O	O
in	NN	O	O
regions	NN	O	O
predominantly	NN	O	O
affected	NN	O	O
in	NN	O	O
MJD	NN	O	B-Disease
.	NN	O	O

Thus	NN	O	O
additional	NN	O	O
molecules	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
regulatory	NN	O	O
events	NN	O	O
are	NN	O	O
necessary	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
exclusive	NN	O	O
degeneration	NN	O	B-Disease
of	NN	O	I-Disease
certain	NN	O	I-Disease
brain	NN	O	I-Disease
areas	NN	O	I-Disease
.	NN	O	O

Emerin	NN	O	B-Disease
,	NN	O	I-Disease
deficiency	NN	O	I-Disease
of	NN	O	O
which	NN	O	O
causes	NN	O	O
Emery	NN	O	B-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
localized	NN	O	O
at	NN	O	O
the	NN	O	O
inner	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EDMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
muscle	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
clinical	NN	O	O
triad	NN	O	O
of	NN	O	O
progressive	NN	O	O
wasting	NN	O	B-Disease
of	NN	O	I-Disease
humero	NN	O	I-Disease
-	NN	O	I-Disease
peroneal	NN	O	I-Disease
muscles	NN	O	I-Disease
,	NN	O	O
early	NN	O	O
contractures	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
elbows	NN	O	I-Disease
,	NN	O	I-Disease
Achilles	NN	O	I-Disease
tendons	NN	O	I-Disease
and	NN	O	I-Disease
postcervical	NN	O	I-Disease
muscles	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
cardiac	NN	O	B-Disease
conduction	NN	O	I-Disease
block	NN	O	I-Disease
with	NN	O	O
a	NN	O	O
high	NN	O	O
risk	NN	O	O
of	NN	O	O
sudden	NN	O	B-Disease
death	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
EDMD	NN	O	B-Disease
on	NN	O	O
Xq28	NN	O	O
encodes	NN	O	O
a	NN	O	O
novel	NN	O	O
protein	NN	O	O
named	NN	O	O
emerin	NN	O	O
that	NN	O	O
localizes	NN	O	O
at	NN	O	O
the	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
of	NN	O	O
skeletal	NN	O	O
,	NN	O	O
cardiac	NN	O	O
and	NN	O	O
smooth	NN	O	O
muscles	NN	O	O
and	NN	O	O
some	NN	O	O
other	NN	O	O
non	NN	O	O
-	NN	O	O
muscle	NN	O	O
tissues	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
a	NN	O	O
possible	NN	O	O
physiological	NN	O	O
role	NN	O	O
for	NN	O	O
emerin	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
ultrastructural	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
in	NN	O	O
human	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
and	NN	O	O
HeLa	NN	O	O
cells	NN	O	O
,	NN	O	O
using	NN	O	O
ultrathin	NN	O	O
cryosections	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
immune	NN	O	O
-	NN	O	O
labeled	NN	O	O
colloidal	NN	O	O
gold	NN	O	O
particles	NN	O	O
were	NN	O	O
localized	NN	O	O
on	NN	O	O
the	NN	O	O
nucleoplasmic	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
inner	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
the	NN	O	O
nuclear	NN	O	O
pore	NN	O	O
.	NN	O	O

Emerin	NN	O	O
stayed	NN	O	O
on	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
lamina	NN	O	O
,	NN	O	O
even	NN	O	O
after	NN	O	O
detergent	NN	O	O
treatment	NN	O	O
that	NN	O	O
solubilizes	NN	O	O
membrane	NN	O	O
lipids	NN	O	O
and	NN	O	O
washes	NN	O	O
out	NN	O	O
membrane	NN	O	O
proteins	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
emerin	NN	O	O
anchors	NN	O	O
at	NN	O	O
the	NN	O	O
inner	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
through	NN	O	O
the	NN	O	O
hydrophobic	NN	O	O
stretch	NN	O	O
,	NN	O	O
and	NN	O	O
protrudes	NN	O	O
from	NN	O	O
the	NN	O	O
hydrophilic	NN	O	O
region	NN	O	O
to	NN	O	O
the	NN	O	O
nucleoplasm	NN	O	O
where	NN	O	O
it	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	O
lamina	NN	O	O
.	NN	O	O

We	NN	O	O
speculate	NN	O	O
that	NN	O	O
emerin	NN	O	O
contributes	NN	O	O
to	NN	O	O
maintain	NN	O	O
the	NN	O	O
nuclear	NN	O	O
structure	NN	O	O
and	NN	O	O
stability	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
nuclear	NN	O	O
functions	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
muscle	NN	O	O
tissues	NN	O	O
that	NN	O	O
have	NN	O	O
severe	NN	O	O
stress	NN	O	O
with	NN	O	O
rigorous	NN	O	O
contraction	NN	O	O
-	NN	O	O
relaxation	NN	O	O
movements	NN	O	O
and	NN	O	O
calcium	NN	O	O
flux	NN	O	O
.	NN	O	O
.	NN	O	O

Locus	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
disease	NN	O	O
locus	NN	O	O
was	NN	O	O
assigned	NN	O	O
to	NN	O	O
chromosome	NN	O	O
9	NN	O	O
and	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
,	NN	O	O
STM7	NN	O	O
/	NN	O	O
X25	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
isolated	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
most	NN	O	O
data	NN	O	O
suggest	NN	O	O
locus	NN	O	O
homogeneity	NN	O	O
in	NN	O	O
FRDA	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
now	NN	O	O
provide	NN	O	O
strong	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
second	NN	O	O
FRDA	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

Studying	NN	O	O
two	NN	O	O
siblings	NN	O	O
with	NN	O	O
FRDA	NN	O	B-Disease
from	NN	O	O
two	NN	O	O
families	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
detect	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
STM7	NN	O	O
/	NN	O	O
X25	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
STM7	NN	O	O
/	NN	O	O
X25	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
9	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
relevant	NN	O	O
portion	NN	O	O
of	NN	O	O
chromosome	NN	O	O
9	NN	O	O
differs	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
patients	NN	O	O
studied	NN	O	O
had	NN	O	O
typical	NN	O	O
FRDA	NN	O	B-Disease
,	NN	O	O
one	NN	O	O
sibpair	NN	O	O
had	NN	O	O
the	NN	O	O
uncommon	NN	O	O
symptom	NN	O	O
of	NN	O	O
retained	NN	O	O
tendon	NN	O	O
reflexes	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
retained	NN	O	O
tendon	NN	O	O
reflexes	NN	O	O
are	NN	O	O
characteristic	NN	O	O
of	NN	O	O
FRDA	NN	O	B-Disease
caused	NN	O	O
by	NN	O	O
the	NN	O	O
second	NN	O	O
locus	NN	O	O
,	NN	O	O
FRDA2	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
an	NN	O	O
unrelated	NN	O	O
FRDA	NN	O	B-Disease
patient	NN	O	O
with	NN	O	O
retained	NN	O	O
tendon	NN	O	O
reflexes	NN	O	O
.	NN	O	O

The	NN	O	O
observation	NN	O	O
of	NN	O	O
typical	NN	O	O
mutations	NN	O	O
in	NN	O	O
STM7	NN	O	O
/	NN	O	O
X25	NN	O	O
(	NN	O	O
GAA	NN	O	O
expansions	NN	O	O
)	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
genetically	NN	O	O
different	NN	O	O
forms	NN	O	O
of	NN	O	O
FRDA	NN	O	B-Disease
cannot	NN	O	O
be	NN	O	O
distinguished	NN	O	O
clinically	NN	O	O
.	NN	O	O
.	NN	O	O

Glycerol	NN	O	O
as	NN	O	O
a	NN	O	O
correlate	NN	O	O
of	NN	O	O
impaired	NN	O	B-Disease
glucose	NN	O	I-Disease
tolerance	NN	O	I-Disease
:	NN	O	O
dissection	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
system	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
simple	NN	O	O
genetic	NN	O	O
trait	NN	O	O
.	NN	O	O

Glycerol	NN	O	O
kinase	NN	O	O
(	NN	O	O
GK	NN	O	O
)	NN	O	O
represents	NN	O	O
the	NN	O	O
primary	NN	O	O
entry	NN	O	O
of	NN	O	O
glycerol	NN	O	O
into	NN	O	O
glucose	NN	O	O
and	NN	O	O
triglyceride	NN	O	O
metabolism	NN	O	O
.	NN	O	O

Impaired	NN	O	B-Disease
glucose	NN	O	I-Disease
tolerance	NN	O	I-Disease
(	NN	O	O
IGT	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
hypertriglyceridemia	NN	O	B-Disease
are	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
diabetes	NN	O	B-Disease
mellitus	NN	O	I-Disease
and	NN	O	O
cardiovascular	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
glycerol	NN	O	O
and	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
IGT	NN	O	B-Disease
,	NN	O	O
however	NN	O	O
,	NN	O	O
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
undertook	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
fasting	NN	O	O
plasma	NN	O	O
glycerol	NN	O	O
levels	NN	O	O
in	NN	O	O
a	NN	O	O
cohort	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
056	NN	O	O
unrelated	NN	O	O
men	NN	O	O
and	NN	O	O
women	NN	O	O
of	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
descent	NN	O	O
.	NN	O	O

Family	NN	O	O
screening	NN	O	O
in	NN	O	O
the	NN	O	O
initial	NN	O	O
cohort	NN	O	O
identified	NN	O	O
18	NN	O	O
men	NN	O	O
from	NN	O	O
five	NN	O	O
families	NN	O	O
with	NN	O	O
severe	NN	O	O
hyperglycerolemia	NN	O	B-Disease
(	NN	O	O
values	NN	O	O
above	NN	O	O
2	NN	O	O
.	NN	O	O
0	NN	O	O
mmol	NN	O	O
/	NN	O	O
liter	NN	O	O
)	NN	O	O
and	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
pattern	NN	O	O
of	NN	O	O
inheritance	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
data	NN	O	O
from	NN	O	O
12	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
surrounding	NN	O	O
the	NN	O	O
Xp21	NN	O	O
.	NN	O	O

3	NN	O	O
GK	NN	O	O
gene	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
peak	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
3	NN	O	O
.	NN	O	O

46	NN	O	O
,	NN	O	O
centered	NN	O	O
around	NN	O	O
marker	NN	O	O
DXS8039	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
since	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
originated	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
with	NN	O	O
a	NN	O	O
proven	NN	O	O
founder	NN	O	O
effect	NN	O	O
-	NN	O	O
the	NN	O	O
Saguenay	NN	O	O
Lac	NN	O	O
-	NN	O	O
St	NN	O	O
.	NN	O	O

-	NN	O	O
Jean	NN	O	O
region	NN	O	O
of	NN	O	O
Quebec	NN	O	O
-	NN	O	O
a	NN	O	O
common	NN	O	O
disease	NN	O	O
haplotype	NN	O	O
was	NN	O	O
sought	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
a	NN	O	O
six	NN	O	O
-	NN	O	O
marker	NN	O	O
haplotype	NN	O	O
extending	NN	O	O
over	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
5	NN	O	O
.	NN	O	O

5	NN	O	O
cM	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
families	NN	O	O
.	NN	O	O

Resequencing	NN	O	O
of	NN	O	O
the	NN	O	O
GK	NN	O	O
gene	NN	O	O
in	NN	O	O
family	NN	O	O
members	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
N288D	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
10	NN	O	O
,	NN	O	O
which	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
asparagine	NN	O	O
residue	NN	O	O
by	NN	O	O
a	NN	O	O
negatively	NN	O	O
charged	NN	O	O
aspartic	NN	O	O
acid	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetics	NN	O	O
of	NN	O	O
primary	NN	O	B-Disease
dystonias	NN	O	I-Disease
.	NN	O	O

Primary	NN	O	B-Disease
dystonias	NN	O	I-Disease
are	NN	O	O
movement	NN	O	B-Disease
disorders	NN	O	I-Disease
with	NN	O	O
dystonia	NN	O	B-Disease
as	NN	O	O
a	NN	O	O
major	NN	O	O
symptom	NN	O	O
.	NN	O	O

They	NN	O	O
are	NN	O	O
frequently	NN	O	O
inherited	NN	O	O
as	NN	O	O
Mendelian	NN	O	O
traits	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
at	NN	O	O
least	NN	O	O
eight	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
and	NN	O	O
two	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
recessive	NN	O	O
forms	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
pedigree	NN	O	O
analyses	NN	O	O
suggest	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
variant	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
classification	NN	O	O
is	NN	O	O
increasingly	NN	O	O
being	NN	O	O
replaced	NN	O	O
by	NN	O	O
a	NN	O	O
genetic	NN	O	O
one	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
gene	NN	O	O
loci	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
six	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
forms	NN	O	O
,	NN	O	O
i	NN	O	O
.	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
in	NN	O	O
idiopathic	NN	O	B-Disease
torsion	NN	O	I-Disease
dystonia	NN	O	I-Disease
(	NN	O	O
9q34	NN	O	O
)	NN	O	O
,	NN	O	O
focal	NN	O	B-Disease
dystonia	NN	O	I-Disease
(	NN	O	O
18p	NN	O	O
)	NN	O	O
,	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
idiopathic	NN	O	B-Disease
torsion	NN	O	I-Disease
dystonia	NN	O	I-Disease
of	NN	O	O
mixed	NN	O	O
type	NN	O	O
(	NN	O	O
8p21	NN	O	O
-	NN	O	O
q22	NN	O	O
)	NN	O	O
,	NN	O	O
dopa	NN	O	B-Disease
-	NN	O	I-Disease
responsive	NN	O	I-Disease
dystonia	NN	O	I-Disease
(	NN	O	O
14q22	NN	O	O
.	NN	O	O
1	NN	O	O
-	NN	O	O
q22	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
paroxysmal	NN	O	B-Disease
dystonic	NN	O	I-Disease
choreoathetosis	NN	O	I-Disease
(	NN	O	O
2q25	NN	O	O
-	NN	O	O
q33	NN	O	O
;	NN	O	O
1p21	NN	O	O
-	NN	O	O
p13	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

Gene	NN	O	O
loci	NN	O	O
in	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
recessive	NN	O	O
forms	NN	O	O
have	NN	O	O
been	NN	O	O
assigned	NN	O	O
to	NN	O	O
Xq13	NN	O	O
.	NN	O	O

1	NN	O	O
in	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
dystonia	NN	O	I-Disease
parkinsonism	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
to	NN	O	O
Xq22	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
sensorineural	NN	O	I-Disease
deafness	NN	O	I-Disease
,	NN	O	O
dystonia	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
disease	NN	O	O
genes	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
two	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
forms	NN	O	O
and	NN	O	O
in	NN	O	O
one	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
recessive	NN	O	O
form	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
a	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
an	NN	O	O
ATP	NN	O	O
-	NN	O	O
binding	NN	O	O
protein	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
idiopathic	NN	O	B-Disease
torsion	NN	O	I-Disease
dystonia	NN	O	I-Disease
(	NN	O	O
DYT1	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
GTP	NN	O	O
cyclohydrolase	NN	O	O
1	NN	O	O
gene	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
dopa	NN	O	B-Disease
-	NN	O	I-Disease
responsive	NN	O	I-Disease
dystonia	NN	O	I-Disease
(	NN	O	O
DYT5	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
sensorineural	NN	O	B-Disease
deafness	NN	O	I-Disease
,	NN	O	O
dystonia	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
DDP	NN	O	O
coding	NN	O	O
for	NN	O	O
a	NN	O	O
polypeptide	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

This	NN	O	O
article	NN	O	O
reviews	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetics	NN	O	O
of	NN	O	O
primary	NN	O	B-Disease
dystonias	NN	O	I-Disease
,	NN	O	O
critically	NN	O	O
discusses	NN	O	O
present	NN	O	O
findings	NN	O	O
,	NN	O	O
and	NN	O	O
proposes	NN	O	O
referring	NN	O	O
to	NN	O	O
the	NN	O	O
known	NN	O	O
forms	NN	O	O
,	NN	O	O
most	NN	O	O
of	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
distinguished	NN	O	O
by	NN	O	O
genetic	NN	O	O
criteria	NN	O	O
,	NN	O	O
as	NN	O	O
dystonias	NN	O	B-Disease
1	NN	O	I-Disease
-	NN	O	I-Disease
12	NN	O	I-Disease
.	NN	O	O

Determination	NN	O	O
of	NN	O	O
30	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
mutations	NN	O	O
,	NN	O	O
including	NN	O	O
15	NN	O	O
not	NN	O	O
previously	NN	O	O
described	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
peroxisomal	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

It	NN	O	O
mainly	NN	O	O
involves	NN	O	O
the	NN	O	O
nervous	NN	O	O
system	NN	O	O
white	NN	O	O
matter	NN	O	O
,	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
and	NN	O	O
testes	NN	O	O
.	NN	O	O

Several	NN	O	O
distinct	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
are	NN	O	O
known	NN	O	O
.	NN	O	O

The	NN	O	O
principal	NN	O	O
biochemical	NN	O	O
abnormality	NN	O	O
is	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
saturated	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
(	NN	O	O
VLCFAs	NN	O	O
>	NN	O	O
C22	NN	O	O
0	NN	O	O
,	NN	O	O
mainly	NN	O	O
C26	NN	O	O
0	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
impaired	NN	O	O
capacity	NN	O	O
for	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
in	NN	O	O
peroxisomes	NN	O	O
.	NN	O	O

Diagnosis	NN	O	O
is	NN	O	O
usually	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
VLCFA	NN	O	O
levels	NN	O	O
in	NN	O	O
plasma	NN	O	O
or	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
in	NN	O	O
both	NN	O	O
patients	NN	O	O
and	NN	O	O
carriers	NN	O	O
.	NN	O	O

In	NN	O	O
0	NN	O	O
.	NN	O	O

1	NN	O	O
%	NN	O	O
of	NN	O	O
affected	NN	O	O
males	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
plasma	NN	O	O
C26	NN	O	O
0	NN	O	O
level	NN	O	O
is	NN	O	O
borderline	NN	O	O
normal	NN	O	O
,	NN	O	O
and	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
obligate	NN	O	O
female	NN	O	O
carriers	NN	O	O
have	NN	O	O
normal	NN	O	O
results	NN	O	O
.	NN	O	O

Effective	NN	O	O
mutation	NN	O	O
detection	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
is	NN	O	O
therefore	NN	O	O
fundamental	NN	O	O
to	NN	O	O
unambiguously	NN	O	O
determine	NN	O	O
the	NN	O	O
genetic	NN	O	O
status	NN	O	O
of	NN	O	O
each	NN	O	O
individual	NN	O	O
at	NN	O	O
risk	NN	O	O
.	NN	O	O

Of	NN	O	O
particular	NN	O	O
concern	NN	O	O
are	NN	O	O
female	NN	O	O
members	NN	O	O
of	NN	O	O
kindreds	NN	O	O
segregating	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
mutations	NN	O	O
,	NN	O	O
because	NN	O	O
normal	NN	O	O
VLCFA	NN	O	O
levels	NN	O	O
do	NN	O	O
not	NN	O	O
guarantee	NN	O	O
lack	NN	O	O
of	NN	O	O
carrier	NN	O	O
status	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
fast	NN	O	O
method	NN	O	O
for	NN	O	O
detection	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
method	NN	O	O
is	NN	O	O
based	NN	O	O
on	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
of	NN	O	O
nested	NN	O	O
PCR	NN	O	O
fragments	NN	O	O
followed	NN	O	O
by	NN	O	O
sequence	NN	O	O
-	NN	O	O
determination	NN	O	O
reactions	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
methodology	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
mutations	NN	O	O
in	NN	O	O
30	NN	O	O
kindreds	NN	O	O
,	NN	O	O
including	NN	O	O
15	NN	O	O
not	NN	O	O
previously	NN	O	O
reported	NN	O	O
.	NN	O	O

beta	NN	O	O
-	NN	O	O
galactosidase	NN	O	O
gene	NN	O	O
mutations	NN	O	O
affecting	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
enzyme	NN	O	O
and	NN	O	O
the	NN	O	O
elastin	NN	O	O
-	NN	O	O
binding	NN	O	O
protein	NN	O	O
in	NN	O	O
GM1	NN	O	B-Disease
-	NN	O	I-Disease
gangliosidosis	NN	O	I-Disease
patients	NN	O	O
with	NN	O	O
cardiac	NN	O	B-Disease
involvement	NN	O	I-Disease
.	NN	O	O

GM1	NN	O	B-Disease
-	NN	O	I-Disease
gangliosidosis	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
lysosomal	NN	O	B-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
acid	NN	O	I-Disease
beta	NN	O	I-Disease
-	NN	O	I-Disease
galactosidase	NN	O	I-Disease
(	NN	O	O
GLB1	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
five	NN	O	O
new	NN	O	O
beta	NN	O	O
-	NN	O	O
galactosidase	NN	O	O
gene	NN	O	O
mutations	NN	O	O
in	NN	O	O
nine	NN	O	O
Italian	NN	O	O
patients	NN	O	O
and	NN	O	O
one	NN	O	O
fetus	NN	O	O
,	NN	O	O
segregating	NN	O	O
in	NN	O	O
seven	NN	O	O
unrelated	NN	O	O
families	NN	O	O
.	NN	O	O

Six	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
infantile	NN	O	O
,	NN	O	O
severe	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
presented	NN	O	O
cardiac	NN	O	B-Disease
involvement	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
feature	NN	O	O
rarely	NN	O	O
associated	NN	O	O
with	NN	O	O
GM1	NN	O	B-Disease
-	NN	O	I-Disease
gangliosidosis	NN	O	I-Disease
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
RNA	NN	O	O
and	NN	O	O
DNA	NN	O	O
identified	NN	O	O
two	NN	O	O
new	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
defects	NN	O	O
,	NN	O	O
three	NN	O	O
new	NN	O	O
and	NN	O	O
three	NN	O	O
previously	NN	O	O
described	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
all	NN	O	O
patients	NN	O	O
with	NN	O	O
cardiac	NN	O	B-Disease
involvement	NN	O	I-Disease
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
R59H	NN	O	O
,	NN	O	O
Y591C	NN	O	O
,	NN	O	O
Y591N	NN	O	O
,	NN	O	O
or	NN	O	O
IVS14	NN	O	O
-	NN	O	O
2A	NN	O	O
>	NN	O	O
G	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
all	NN	O	O
other	NN	O	O
patients	NN	O	O
were	NN	O	O
compound	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
following	NN	O	O
mutations	NN	O	O
R201H	NN	O	O
,	NN	O	O
R482H	NN	O	O
,	NN	O	O
G579D	NN	O	O
,	NN	O	O
IVS8	NN	O	O
+	NN	O	O
2T	NN	O	O
>	NN	O	O
C	NN	O	O
.	NN	O	O

Although	NN	O	O
we	NN	O	O
could	NN	O	O
not	NN	O	O
directly	NN	O	O
correlate	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cardiac	NN	O	B-Disease
abnormalities	NN	O	I-Disease
with	NN	O	O
specific	NN	O	O
genetic	NN	O	B-Disease
lesions	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
mutations	NN	O	O
identified	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
cardiomyopathy	NN	O	B-Disease
fell	NN	O	O
in	NN	O	O
the	NN	O	O
GLB1	NN	O	O
cDNA	NN	O	O
region	NN	O	O
common	NN	O	O
to	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
enzyme	NN	O	O
and	NN	O	O
the	NN	O	O
Hbeta	NN	O	O
-	NN	O	O
Gal	NN	O	O
-	NN	O	O
related	NN	O	O
protein	NN	O	O
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
the	NN	O	O
elastin	NN	O	O
binding	NN	O	O
protein	NN	O	O
(	NN	O	O
EBP	NN	O	O
)	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
both	NN	O	O
molecules	NN	O	O
are	NN	O	O
affected	NN	O	O
by	NN	O	O
the	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
may	NN	O	O
contribute	NN	O	O
differently	NN	O	O
to	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
specific	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
.	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
modulation	NN	O	O
of	NN	O	O
Hmgic	NN	O	O
reduces	NN	O	O
obesity	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
HMGI	NN	O	O
family	NN	O	O
of	NN	O	O
proteins	NN	O	O
consists	NN	O	O
of	NN	O	O
three	NN	O	O
members	NN	O	O
,	NN	O	O
HMGIC	NN	O	O
,	NN	O	O
HMGI	NN	O	O
and	NN	O	O
HMGI	NN	O	O
(	NN	O	O
Y	NN	O	O
)	NN	O	O
,	NN	O	O
that	NN	O	O
function	NN	O	O
as	NN	O	O
architectural	NN	O	O
factors	NN	O	O
and	NN	O	O
are	NN	O	O
essential	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
enhancesome	NN	O	O
.	NN	O	O

HMGIC	NN	O	O
is	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
proliferating	NN	O	O
,	NN	O	O
undifferentiated	NN	O	O
mesenchymal	NN	O	O
cells	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
adult	NN	O	O
tissues	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
disrupted	NN	O	O
and	NN	O	O
misexpressed	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
mesenchymal	NN	O	B-Disease
tumour	NN	O	I-Disease
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
fat	NN	O	B-Disease
-	NN	O	I-Disease
cell	NN	O	I-Disease
tumours	NN	O	I-Disease
(	NN	O	O
lipomas	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
Hmgic	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
have	NN	O	O
a	NN	O	O
deficiency	NN	O	O
in	NN	O	O
fat	NN	O	O
tissue	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
adipogenesis	NN	O	O
and	NN	O	O
obesity	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
examined	NN	O	O
Hmgic	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
adipose	NN	O	O
tissue	NN	O	O
of	NN	O	O
adult	NN	O	O
,	NN	O	O
obese	NN	O	B-Disease
mice	NN	O	O
.	NN	O	O

Mice	NN	O	O
with	NN	O	O
a	NN	O	O
partial	NN	O	B-Disease
or	NN	O	I-Disease
complete	NN	O	I-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
Hmgic	NN	O	I-Disease
resisted	NN	O	O
diet	NN	O	O
-	NN	O	O
induced	NN	O	O
obesity	NN	O	B-Disease
.	NN	O	O

Disruption	NN	O	O
of	NN	O	O
Hmgic	NN	O	O
caused	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
obesity	NN	O	B-Disease
induced	NN	O	O
by	NN	O	O
leptin	NN	O	B-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
Lepob	NN	O	O
/	NN	O	O
Lepob	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
gene	NN	O	O
-	NN	O	O
dose	NN	O	O
-	NN	O	O
dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
implicate	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
HMGIC	NN	O	O
in	NN	O	O
fat	NN	O	O
-	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
adipose	NN	O	O
-	NN	O	O
specific	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
obesity	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
relationship	NN	O	O
in	NN	O	O
factor	NN	O	B-Disease
X	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Factor	NN	O	B-Disease
X	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
haemorrhagic	NN	O	B-Disease
condition	NN	O	I-Disease
,	NN	O	O
normally	NN	O	O
inherited	NN	O	O
as	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
trait	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
a	NN	O	O
variable	NN	O	O
clinical	NN	O	O
presentation	NN	O	O
correlates	NN	O	O
poorly	NN	O	O
with	NN	O	O
laboratory	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
factor	NN	O	O
X	NN	O	O
(	NN	O	O
F10	NN	O	O
)	NN	O	O
genes	NN	O	O
of	NN	O	O
14	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
with	NN	O	O
factor	NN	O	B-Disease
X	NN	O	I-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
12	NN	O	O
familial	NN	O	O
and	NN	O	O
two	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
)	NN	O	O
were	NN	O	O
sequenced	NN	O	O
yielding	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
13	NN	O	O
novel	NN	O	O
mutations	NN	O	O
.	NN	O	O

Family	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
distinguish	NN	O	O
the	NN	O	O
contributions	NN	O	O
of	NN	O	O
individual	NN	O	O
mutant	NN	O	O
F10	NN	O	O
alleles	NN	O	O
to	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
laboratory	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
studied	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
molecular	NN	O	O
modelling	NN	O	O
,	NN	O	O
whereas	NN	O	O
single	NN	O	O
basepair	NN	O	O
substitutions	NN	O	O
in	NN	O	O
splice	NN	O	O
sites	NN	O	O
and	NN	O	O
the	NN	O	O
5	NN	O	O
flanking	NN	O	O
region	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
splicing	NN	O	O
assay	NN	O	O
and	NN	O	O
luciferase	NN	O	O
reporter	NN	O	O
gene	NN	O	O
assay	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
allele	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
hexanucleotide	NN	O	O
insertion	NN	O	O
/	NN	O	O
deletion	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
the	NN	O	O
F10	NN	O	O
gene	NN	O	O
promoter	NN	O	O
region	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
reporter	NN	O	O
gene	NN	O	O
assay	NN	O	O
,	NN	O	O
to	NN	O	O
reduce	NN	O	O
promoter	NN	O	O
activity	NN	O	O
by	NN	O	O
approximately	NN	O	O
20	NN	O	O
%	NN	O	O
.	NN	O	O

One	NN	O	O
family	NN	O	O
manifesting	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
pattern	NN	O	O
of	NN	O	O
inheritance	NN	O	O
possessed	NN	O	O
three	NN	O	O
clinically	NN	O	O
affected	NN	O	O
members	NN	O	O
who	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
that	NN	O	O
was	NN	O	O
predicted	NN	O	O
to	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
protein	NN	O	O
product	NN	O	O
.	NN	O	O

A	NN	O	O
model	NN	O	O
which	NN	O	O
accounts	NN	O	O
for	NN	O	O
the	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
lesion	NN	O	O
is	NN	O	O
presented	NN	O	O
.	NN	O	O

Variation	NN	O	O
in	NN	O	O
the	NN	O	O
antigen	NN	O	O
level	NN	O	O
of	NN	O	O
heterozygous	NN	O	O
relatives	NN	O	O
of	NN	O	O
probands	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
significantly	NN	O	O
higher	NN	O	O
between	NN	O	O
families	NN	O	O
than	NN	O	O
within	NN	O	O
families	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
F10	NN	O	O
lesion	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
segregating	NN	O	O
in	NN	O	O
a	NN	O	O
given	NN	O	O
family	NN	O	O
is	NN	O	O
a	NN	O	O
prime	NN	O	O
determinant	NN	O	O
of	NN	O	O
the	NN	O	O
laboratory	NN	O	O
phenotype	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
no	NN	O	O
such	NN	O	O
relationship	NN	O	O
could	NN	O	O
be	NN	O	O
discerned	NN	O	O
between	NN	O	O
laboratory	NN	O	O
phenotype	NN	O	O
and	NN	O	O
polymorphism	NN	O	O
genotype	NN	O	O
.	NN	O	O
.	NN	O	O

Transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
a	NN	O	O
truncated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
high	NN	O	O
mobility	NN	O	O
group	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
protein	NN	O	O
develop	NN	O	O
adiposity	NN	O	O
and	NN	O	O
an	NN	O	O
abnormally	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
lipomas	NN	O	B-Disease
.	NN	O	O

Chromosomal	NN	O	O
translocations	NN	O	O
in	NN	O	O
human	NN	O	O
lipomas	NN	O	B-Disease
frequently	NN	O	O
create	NN	O	O
fusion	NN	O	O
transcripts	NN	O	O
encoding	NN	O	O
high	NN	O	O
mobility	NN	O	O
group	NN	O	O
(	NN	O	O
HMG	NN	O	O
)	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
domains	NN	O	O
and	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
sequences	NN	O	O
from	NN	O	O
different	NN	O	O
presumed	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
for	NN	O	O
HMG	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lipomas	NN	O	B-Disease
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
HMG	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
component	NN	O	O
,	NN	O	O
the	NN	O	O
three	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
domains	NN	O	O
of	NN	O	O
HMG	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
expressed	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
ubiquitous	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
truncated	NN	O	O
HMG	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
protein	NN	O	O
,	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
develop	NN	O	O
a	NN	O	O
selective	NN	O	O
abundance	NN	O	O
of	NN	O	O
fat	NN	O	O
tissue	NN	O	O
early	NN	O	O
in	NN	O	O
life	NN	O	O
,	NN	O	O
show	NN	O	O
marked	NN	O	O
adipose	NN	O	B-Disease
tissue	NN	O	I-Disease
inflammation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
have	NN	O	O
an	NN	O	O
abnormally	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
lipomas	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
domains	NN	O	O
of	NN	O	O
HMG	NN	O	O
I	NN	O	O
-	NN	O	O
C	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
fusion	NN	O	O
partner	NN	O	O
,	NN	O	O
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
perturb	NN	O	O
adipogenesis	NN	O	O
and	NN	O	O
predispose	NN	O	O
to	NN	O	O
lipomas	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
provide	NN	O	O
data	NN	O	O
supporting	NN	O	O
the	NN	O	O
central	NN	O	O
utility	NN	O	O
of	NN	O	O
this	NN	O	O
animal	NN	O	O
model	NN	O	O
as	NN	O	O
a	NN	O	O
tool	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
kind	NN	O	O
of	NN	O	O
human	NN	O	O
benign	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

ATM	NN	O	O
phosphorylates	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
in	NN	O	O
an	NN	O	O
S	NN	O	O
-	NN	O	O
phase	NN	O	O
checkpoint	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
rare	NN	O	B-Disease
diseases	NN	O	I-Disease
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
AT	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ATM	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
Nijmegen	NN	O	B-Disease
breakage	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
NBS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
gene	NN	O	O
,	NN	O	O
share	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
phenotypic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
such	NN	O	O
as	NN	O	O
chromosomal	NN	O	B-Disease
instability	NN	O	I-Disease
,	NN	O	O
radiation	NN	O	O
sensitivity	NN	O	O
and	NN	O	O
defects	NN	O	O
in	NN	O	O
cell	NN	O	O
-	NN	O	O
cycle	NN	O	O
checkpoints	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

The	NN	O	O
ATM	NN	O	O
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
kinase	NN	O	O
that	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
or	NN	O	O
radiomimetic	NN	O	O
drugs	NN	O	O
,	NN	O	O
whereas	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
complex	NN	O	O
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
responses	NN	O	O
to	NN	O	O
DNA	NN	O	O
double	NN	O	O
-	NN	O	O
strand	NN	O	O
breaks	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
similarities	NN	O	O
between	NN	O	O
AT	NN	O	B-Disease
and	NN	O	O
NBS	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
functional	NN	O	O
interactions	NN	O	O
between	NN	O	O
ATM	NN	O	O
and	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
ATM	NN	O	O
kinase	NN	O	O
by	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
ATM	NN	O	O
-	NN	O	O
dependent	NN	O	O
responses	NN	O	O
in	NN	O	O
NBS	NN	O	B-Disease
cells	NN	O	O
indicated	NN	O	O
that	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
signalling	NN	O	O
to	NN	O	O
ATM	NN	O	O
after	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
was	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
serine	NN	O	O
343	NN	O	O
in	NN	O	O
an	NN	O	O
ATM	NN	O	O
-	NN	O	O
dependent	NN	O	O
manner	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
after	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

A	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
construct	NN	O	O
mutated	NN	O	O
at	NN	O	O
the	NN	O	O
ATM	NN	O	O
phosphorylation	NN	O	O
site	NN	O	O
abrogated	NN	O	O
an	NN	O	O
S	NN	O	O
-	NN	O	O
phase	NN	O	O
checkpoint	NN	O	O
induced	NN	O	O
by	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
in	NN	O	O
normal	NN	O	O
cells	NN	O	O
and	NN	O	O
failed	NN	O	O
to	NN	O	O
compensate	NN	O	O
for	NN	O	O
this	NN	O	O
functional	NN	O	O
deficiency	NN	O	O
in	NN	O	O
NBS	NN	O	B-Disease
cells	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
link	NN	O	O
ATM	NN	O	O
and	NN	O	O
p95	NN	O	O
/	NN	O	O
nbs1	NN	O	O
in	NN	O	O
a	NN	O	O
common	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
and	NN	O	O
provide	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
phenotypic	NN	O	O
similarities	NN	O	O
in	NN	O	O
these	NN	O	O
two	NN	O	O
diseases	NN	O	O
.	NN	O	O
.	NN	O	O

Understanding	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
inherited	NN	O	B-Disease
mental	NN	O	I-Disease
retardation	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
mainly	NN	O	O
caused	NN	O	O
by	NN	O	O
massive	NN	O	O
expansion	NN	O	O
of	NN	O	O
CGG	NN	O	O
triplet	NN	O	O
repeats	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
-	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
mental	NN	O	I-Disease
retardation	NN	O	I-Disease
-	NN	O	O
1	NN	O	O
(	NN	O	O
FMR1	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
with	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
expanded	NN	O	O
CGG	NN	O	O
triplet	NN	O	O
repeats	NN	O	O
are	NN	O	O
hypermethylated	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
is	NN	O	O
repressed	NN	O	O
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
FMR1	NN	O	O
protein	NN	O	O
(	NN	O	O
FMRP	NN	O	O
)	NN	O	O
and	NN	O	O
subsequent	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
.	NN	O	O

FMRP	NN	O	O
is	NN	O	O
an	NN	O	O
RNA	NN	O	O
-	NN	O	O
binding	NN	O	O
protein	NN	O	O
that	NN	O	O
shuttles	NN	O	O
between	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

This	NN	O	O
protein	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
protein	NN	O	O
translation	NN	O	O
as	NN	O	O
it	NN	O	O
is	NN	O	O
found	NN	O	O
associated	NN	O	O
with	NN	O	O
polyribosomes	NN	O	O
and	NN	O	O
the	NN	O	O
rough	NN	O	O
endoplasmic	NN	O	O
reticulum	NN	O	O
.	NN	O	O

We	NN	O	O
discuss	NN	O	O
here	NN	O	O
the	NN	O	O
recent	NN	O	O
progress	NN	O	O
made	NN	O	O
towards	NN	O	O
understanding	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
CGG	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
and	NN	O	O
physiological	NN	O	O
function	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
FMRP	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
will	NN	O	O
not	NN	O	O
only	NN	O	O
help	NN	O	O
to	NN	O	O
illuminate	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
class	NN	O	O
of	NN	O	O
human	NN	O	O
diseases	NN	O	O
with	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
but	NN	O	O
also	NN	O	O
provide	NN	O	O
an	NN	O	O
avenue	NN	O	O
to	NN	O	O
understand	NN	O	O
aspects	NN	O	O
of	NN	O	O
human	NN	O	O
cognition	NN	O	O
and	NN	O	O
intelligence	NN	O	O
.	NN	O	O
.	NN	O	O

Haploinsufficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
transcription	NN	O	I-Disease
factors	NN	O	I-Disease
FOXC1	NN	O	I-Disease
and	NN	O	I-Disease
FOXC2	NN	O	I-Disease
results	NN	O	O
in	NN	O	O
aberrant	NN	O	O
ocular	NN	O	O
development	NN	O	O
.	NN	O	O

Anterior	NN	O	B-Disease
segment	NN	O	I-Disease
developmental	NN	O	I-Disease
disorders	NN	O	I-Disease
,	NN	O	O
including	NN	O	O
Axenfeld	NN	O	B-Disease
-	NN	O	I-Disease
Rieger	NN	O	I-Disease
anomaly	NN	O	I-Disease
(	NN	O	O
ARA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
variably	NN	O	O
associate	NN	O	O
with	NN	O	O
harmfully	NN	O	O
elevated	NN	O	O
intraocular	NN	O	O
pressure	NN	O	O
(	NN	O	O
IOP	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
causes	NN	O	O
glaucoma	NN	O	B-Disease
.	NN	O	O

Clinically	NN	O	O
observed	NN	O	O
dysgenesis	NN	O	O
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
IOP	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
etiology	NN	O	O
of	NN	O	O
glaucoma	NN	O	B-Disease
development	NN	O	O
is	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
forkhead	NN	O	O
transcription	NN	O	O
factor	NN	O	O
genes	NN	O	O
Foxc1	NN	O	O
(	NN	O	O
formerly	NN	O	O
Mf1	NN	O	O
)	NN	O	O
and	NN	O	O
Foxc2	NN	O	O
(	NN	O	O
formerly	NN	O	O
Mfh1	NN	O	O
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
mesenchyme	NN	O	O
from	NN	O	O
which	NN	O	O
the	NN	O	O
ocular	NN	O	O
drainage	NN	O	O
structures	NN	O	O
derive	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
of	NN	O	O
Foxc1	NN	O	O
,	NN	O	O
FKHL7	NN	O	O
,	NN	O	O
cause	NN	O	O
dominant	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
defects	NN	O	I-Disease
and	NN	O	O
glaucoma	NN	O	B-Disease
in	NN	O	O
various	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
Foxc1	NN	O	O
(	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
)	NN	O	O
mice	NN	O	O
have	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
abnormalities	NN	O	I-Disease
similar	NN	O	O
to	NN	O	O
those	NN	O	O
reported	NN	O	O
in	NN	O	O
human	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
abnormalities	NN	O	O
include	NN	O	O
small	NN	O	O
or	NN	O	O
absent	NN	O	O
Schlemms	NN	O	O
canal	NN	O	O
,	NN	O	O
aberrantly	NN	O	O
developed	NN	O	O
trabecular	NN	O	O
meshwork	NN	O	O
,	NN	O	O
iris	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
severely	NN	O	O
eccentric	NN	O	O
pupils	NN	O	O
and	NN	O	O
displaced	NN	O	O
Schwalbes	NN	O	O
line	NN	O	O
.	NN	O	O

The	NN	O	O
penetrance	NN	O	O
of	NN	O	O
clinically	NN	O	O
obvious	NN	O	O
abnormalities	NN	O	O
varies	NN	O	O
with	NN	O	O
genetic	NN	O	O
background	NN	O	O
.	NN	O	O

In	NN	O	O
some	NN	O	O
affected	NN	O	O
eyes	NN	O	O
,	NN	O	O
collagen	NN	O	O
bundles	NN	O	O
were	NN	O	O
half	NN	O	O
normal	NN	O	O
diameter	NN	O	O
,	NN	O	O
or	NN	O	O
collagen	NN	O	O
and	NN	O	O
elastic	NN	O	O
tissue	NN	O	O
were	NN	O	O
very	NN	O	O
sparse	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
synthesis	NN	O	O
or	NN	O	O
organization	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
ocular	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
ocular	NN	O	O
drainage	NN	O	O
structures	NN	O	O
in	NN	O	O
Foxc1	NN	O	O
(	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
)	NN	O	O
mice	NN	O	O
,	NN	O	O
IOP	NN	O	O
was	NN	O	O
normal	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
mice	NN	O	O
analyzed	NN	O	O
,	NN	O	O
on	NN	O	O
all	NN	O	O
genetic	NN	O	O
backgrounds	NN	O	O
and	NN	O	O
at	NN	O	O
all	NN	O	O
ages	NN	O	O
.	NN	O	O

Similar	NN	O	O
abnormalities	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
Foxc2	NN	O	O
(	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
)	NN	O	O
mice	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
disease	NN	O	O
-	NN	O	O
associated	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
FKHL14	NN	O	O
in	NN	O	O
32	NN	O	O
ARA	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Foxc1	NN	O	O
(	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
)	NN	O	O
and	NN	O	O
Foxc2	NN	O	O
(	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
)	NN	O	O
mice	NN	O	O
are	NN	O	O
useful	NN	O	O
models	NN	O	O
for	NN	O	O
studying	NN	O	O
anterior	NN	O	O
segment	NN	O	O
development	NN	O	O
and	NN	O	O
its	NN	O	O
anomalies	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
allow	NN	O	O
identification	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
Foxc1	NN	O	O
and	NN	O	O
Foxc2	NN	O	O
(	NN	O	O
or	NN	O	O
FKHL7	NN	O	O
and	NN	O	O
FKHL14	NN	O	O
)	NN	O	O
to	NN	O	O
produce	NN	O	O
a	NN	O	O
phenotype	NN	O	O
with	NN	O	O
elevated	NN	O	O
IOP	NN	O	O
and	NN	O	O
glaucoma	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

(	NN	O	O
Over	NN	O	O
)	NN	O	O
correction	NN	O	O
of	NN	O	O
FMR1	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
YAC	NN	O	O
transgenics	NN	O	O
:	NN	O	O
behavioral	NN	O	O
and	NN	O	O
physical	NN	O	O
features	NN	O	O
.	NN	O	O

Fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
common	NN	O	O
cause	NN	O	O
of	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
involving	NN	O	O
loss	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
FMR1	NN	O	O
remains	NN	O	O
undetermined	NN	O	O
but	NN	O	O
the	NN	O	O
protein	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
RNA	NN	O	O
metabolism	NN	O	O
.	NN	O	O

Fmr1	NN	O	O
knockout	NN	O	O
mice	NN	O	O
exhibit	NN	O	O
a	NN	O	O
phenotype	NN	O	O
with	NN	O	O
some	NN	O	O
similarities	NN	O	O
to	NN	O	O
humans	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
macroorchidism	NN	O	B-Disease
and	NN	O	O
behavioral	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
step	NN	O	O
toward	NN	O	O
understanding	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
FMR1	NN	O	O
and	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
the	NN	O	O
potential	NN	O	O
for	NN	O	O
therapeutic	NN	O	O
approaches	NN	O	O
to	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosome	NN	O	O
(	NN	O	O
YAC	NN	O	O
)	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
were	NN	O	O
generated	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
Fmr1	NN	O	O
knockout	NN	O	O
mouse	NN	O	O
phenotype	NN	O	O
could	NN	O	O
be	NN	O	O
rescued	NN	O	O
.	NN	O	O

Several	NN	O	O
transgenic	NN	O	O
lines	NN	O	O
were	NN	O	O
generated	NN	O	O
that	NN	O	O
carried	NN	O	O
the	NN	O	O
entire	NN	O	O
FMR1	NN	O	O
locus	NN	O	O
with	NN	O	O
extensive	NN	O	O
amounts	NN	O	O
of	NN	O	O
flanking	NN	O	O
sequence	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
the	NN	O	O
YAC	NN	O	O
transgene	NN	O	O
supported	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
protein	NN	O	O
(	NN	O	O
FMRP	NN	O	O
)	NN	O	O
which	NN	O	O
was	NN	O	O
present	NN	O	O
at	NN	O	O
levels	NN	O	O
10	NN	O	O
to	NN	O	O
15	NN	O	O
times	NN	O	O
that	NN	O	O
of	NN	O	O
endogenous	NN	O	O
protein	NN	O	O
and	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
-	NN	O	O
and	NN	O	O
tissue	NN	O	O
-	NN	O	O
specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Macro	NN	O	O
-	NN	O	O
orchidism	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
knockout	NN	O	O
mice	NN	O	O
carrying	NN	O	O
the	NN	O	O
YAC	NN	O	O
transgene	NN	O	O
indicating	NN	O	O
functional	NN	O	O
rescue	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	O
protein	NN	O	O
.	NN	O	O

Given	NN	O	O
the	NN	O	O
complex	NN	O	O
behavioral	NN	O	O
phenotype	NN	O	O
in	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
patients	NN	O	O
and	NN	O	O
the	NN	O	O
mild	NN	O	O
phenotype	NN	O	O
previously	NN	O	O
reported	NN	O	O
for	NN	O	O
the	NN	O	O
Fmr1	NN	O	O
knockout	NN	O	O
mouse	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
a	NN	O	O
more	NN	O	O
thorough	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
Fmr1	NN	O	O
knockout	NN	O	O
phenotype	NN	O	O
using	NN	O	O
additional	NN	O	O
behavioral	NN	O	O
assays	NN	O	O
that	NN	O	O
had	NN	O	O
not	NN	O	O
previously	NN	O	O
been	NN	O	O
reported	NN	O	O
for	NN	O	O
this	NN	O	O
animal	NN	O	O
model	NN	O	O
.	NN	O	O

The	NN	O	O
mouse	NN	O	O
displayed	NN	O	O
reduced	NN	O	O
anxiety	NN	O	B-Disease
-	NN	O	O
related	NN	O	O
responses	NN	O	O
with	NN	O	O
increased	NN	O	O
exploratory	NN	O	O
behavior	NN	O	O
.	NN	O	O

FMR1	NN	O	O
YAC	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
overexpressing	NN	O	O
the	NN	O	O
human	NN	O	O
protein	NN	O	O
did	NN	O	O
produce	NN	O	O
opposing	NN	O	O
behavioral	NN	O	O
responses	NN	O	O
and	NN	O	O
additional	NN	O	O
abnormal	NN	O	O
behaviors	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
significant	NN	O	O
implications	NN	O	O
for	NN	O	O
gene	NN	O	O
therapy	NN	O	O
for	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
since	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
may	NN	O	O
harbor	NN	O	O
its	NN	O	O
own	NN	O	O
phenotype	NN	O	O
.	NN	O	O
.	NN	O	O

Transgenic	NN	O	O
mice	NN	O	O
carrying	NN	O	O
large	NN	O	O
human	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
with	NN	O	O
expanded	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
mimic	NN	O	O
closely	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
CTG	NN	O	O
repeat	NN	O	O
intergenerational	NN	O	O
and	NN	O	O
somatic	NN	O	O
instability	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
in	NN	O	O
the	NN	O	O
3UTR	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
DMPK	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

A	NN	O	O
very	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
instability	NN	O	O
is	NN	O	O
observed	NN	O	O
through	NN	O	O
successive	NN	O	O
generations	NN	O	O
and	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
is	NN	O	O
generally	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
and	NN	O	O
with	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
tissues	NN	O	O
from	NN	O	O
DM	NN	O	B-Disease
patients	NN	O	O
exhibit	NN	O	O
somatic	NN	O	O
mosaicism	NN	O	O
that	NN	O	O
increases	NN	O	O
with	NN	O	O
age	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
carrying	NN	O	O
large	NN	O	O
human	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
with	NN	O	O
20	NN	O	O
,	NN	O	O
55	NN	O	O
or	NN	O	O
>	NN	O	O
300	NN	O	O
CTG	NN	O	O
,	NN	O	O
cloned	NN	O	O
from	NN	O	O
patients	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
affected	NN	O	O
DM	NN	O	B-Disease
family	NN	O	O
.	NN	O	O

Using	NN	O	O
large	NN	O	O
human	NN	O	O
flanking	NN	O	O
sequences	NN	O	O
and	NN	O	O
a	NN	O	O
large	NN	O	O
amplification	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
intergenerational	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
instability	NN	O	O
is	NN	O	O
reproduced	NN	O	O
in	NN	O	O
mice	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
strong	NN	O	O
bias	NN	O	O
towards	NN	O	O
expansions	NN	O	O
and	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
sex	NN	O	O
-	NN	O	O
and	NN	O	O
size	NN	O	O
-	NN	O	O
dependent	NN	O	O
characteristics	NN	O	O
as	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
instability	NN	O	O
,	NN	O	O
increasing	NN	O	O
with	NN	O	O
age	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
observed	NN	O	O
in	NN	O	O
tissues	NN	O	O
and	NN	O	O
in	NN	O	O
sperm	NN	O	O
.	NN	O	O

Although	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
observe	NN	O	O
dramatic	NN	O	O
expansions	NN	O	O
(	NN	O	O
or	NN	O	O
big	NN	O	O
jumps	NN	O	O
over	NN	O	O
several	NN	O	O
hundred	NN	O	O
CTG	NN	O	O
repeats	NN	O	O
)	NN	O	O
as	NN	O	O
in	NN	O	O
congenital	NN	O	O
forms	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
,	NN	O	O
our	NN	O	O
model	NN	O	O
carrying	NN	O	O
>	NN	O	O
300	NN	O	O
CTG	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
show	NN	O	O
instability	NN	O	O
so	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
DM	NN	O	B-Disease
situation	NN	O	O
.	NN	O	O

Our	NN	O	O
three	NN	O	O
models	NN	O	O
carrying	NN	O	O
different	NN	O	O
sizes	NN	O	O
of	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
provide	NN	O	O
insight	NN	O	O
on	NN	O	O
the	NN	O	O
different	NN	O	O
factors	NN	O	O
modulating	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
instability	NN	O	O
.	NN	O	O
.	NN	O	O

Inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
mouse	NN	O	O
gene	NN	O	O
leads	NN	O	O
to	NN	O	O
early	NN	O	O
embryonic	NN	O	B-Disease
lethality	NN	O	I-Disease
without	NN	O	O
iron	NN	O	O
accumulation	NN	O	O
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
ataxia	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
caused	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
cases	NN	O	O
by	NN	O	O
homozygous	NN	O	O
intronic	NN	O	O
expansions	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
frataxin	NN	O	O
,	NN	O	O
a	NN	O	O
mitochondrial	NN	O	O
protein	NN	O	O
conserved	NN	O	O
through	NN	O	O
evolution	NN	O	O
,	NN	O	O
and	NN	O	O
involved	NN	O	O
in	NN	O	O
mitochondrial	NN	O	O
iron	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Yeast	NN	O	O
knockout	NN	O	O
models	NN	O	O
,	NN	O	O
and	NN	O	O
histological	NN	O	O
and	NN	O	O
biochemical	NN	O	O
data	NN	O	O
from	NN	O	O
patient	NN	O	O
heart	NN	O	O
biopsies	NN	O	O
or	NN	O	O
autopsies	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
frataxin	NN	O	O
defect	NN	O	O
causes	NN	O	O
a	NN	O	O
specific	NN	O	O
iron	NN	O	B-Disease
-	NN	O	I-Disease
sulfur	NN	O	I-Disease
protein	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
mitochondrial	NN	O	O
iron	NN	O	O
accumulation	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
pathological	NN	O	O
changes	NN	O	O
.	NN	O	O

Affected	NN	O	O
human	NN	O	O
tissues	NN	O	O
are	NN	O	O
rarely	NN	O	O
available	NN	O	O
to	NN	O	O
further	NN	O	O
examine	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
mouse	NN	O	O
model	NN	O	O
by	NN	O	O
deletion	NN	O	O
of	NN	O	O
exon	NN	O	O
4	NN	O	O
leading	NN	O	O
to	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
Frda	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
homozygous	NN	O	O
deletions	NN	O	O
cause	NN	O	O
embryonic	NN	O	B-Disease
lethality	NN	O	I-Disease
a	NN	O	O
few	NN	O	O
days	NN	O	O
after	NN	O	O
implantation	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
frataxin	NN	O	O
during	NN	O	O
early	NN	O	O
development	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
milder	NN	O	O
phenotype	NN	O	O
in	NN	O	O
humans	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
residual	NN	O	O
frataxin	NN	O	O
expression	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
expansion	NN	O	O
mutations	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
frataxin	NN	O	O
knockout	NN	O	O
mouse	NN	O	O
,	NN	O	O
no	NN	O	O
iron	NN	O	O
accumulation	NN	O	O
was	NN	O	O
observed	NN	O	O
during	NN	O	O
embryonic	NN	O	O
resorption	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
cell	NN	O	O
death	NN	O	O
could	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
mechanism	NN	O	O
independent	NN	O	O
of	NN	O	O
iron	NN	O	O
accumulation	NN	O	O
.	NN	O	O
.	NN	O	O

Gaucher	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
the	NN	O	O
origins	NN	O	O
of	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
N370S	NN	O	O
and	NN	O	O
84GG	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
glucosidase	NN	O	O
mutations	NN	O	O
.	NN	O	O

Type	NN	O	O
1	NN	O	O
Gaucher	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
GD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
non	NN	O	O
-	NN	O	O
neuronopathic	NN	O	O
lysosomal	NN	O	B-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
results	NN	O	O
from	NN	O	O
the	NN	O	O
deficient	NN	O	O
activity	NN	O	O
of	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
glucosidase	NN	O	O
(	NN	O	O
GBA	NN	O	O
)	NN	O	O
.	NN	O	O

Type	NN	O	O
1	NN	O	O
disease	NN	O	O
is	NN	O	O
panethnic	NN	O	O
but	NN	O	O
is	NN	O	O
more	NN	O	O
prevalent	NN	O	O
in	NN	O	O
individuals	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
(	NN	O	O
AJ	NN	O	O
)	NN	O	O
descent	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
causative	NN	O	O
GBA	NN	O	O
mutations	NN	O	O
,	NN	O	O
N370S	NN	O	O
is	NN	O	O
particularly	NN	O	O
frequent	NN	O	O
in	NN	O	O
the	NN	O	O
AJ	NN	O	O
population	NN	O	O
,	NN	O	O
(	NN	O	O
q	NN	O	O
approximately	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
84GG	NN	O	O
insertion	NN	O	O
(	NN	O	O
q	NN	O	O
approximately	NN	O	O
.	NN	O	O
003	NN	O	O
)	NN	O	O
occurs	NN	O	O
exclusively	NN	O	O
in	NN	O	O
the	NN	O	O
Ashkenazim	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
genetic	NN	O	O
history	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AJ	NN	O	O
population	NN	O	O
,	NN	O	O
short	NN	O	O
tandem	NN	O	O
repeat	NN	O	O
(	NN	O	O
STR	NN	O	O
)	NN	O	O
markers	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
map	NN	O	O
a	NN	O	O
9	NN	O	O
.	NN	O	O

3	NN	O	O
-	NN	O	O
cM	NN	O	O
region	NN	O	O
containing	NN	O	O
the	NN	O	O
GBA	NN	O	O
locus	NN	O	O
and	NN	O	O
to	NN	O	O
genotype	NN	O	O
261	NN	O	O
AJ	NN	O	O
N370S	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
60	NN	O	O
European	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
N370S	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
and	NN	O	O
62	NN	O	O
AJ	NN	O	O
84GG	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

A	NN	O	O
highly	NN	O	O
conserved	NN	O	O
haplotype	NN	O	O
at	NN	O	O
four	NN	O	O
markers	NN	O	O
flanking	NN	O	O
GBA	NN	O	O
(	NN	O	O
PKLR	NN	O	O
,	NN	O	O
D1S1595	NN	O	O
,	NN	O	O
D1S2721	NN	O	O
,	NN	O	O
and	NN	O	O
D1S2777	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
on	NN	O	O
both	NN	O	O
the	NN	O	O
AJ	NN	O	O
chromosomes	NN	O	O
and	NN	O	O
the	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
N370S	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
a	NN	O	O
founder	NN	O	O
common	NN	O	O
to	NN	O	O
both	NN	O	O
populations	NN	O	O
.	NN	O	O

Of	NN	O	O
note	NN	O	O
,	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
different	NN	O	O
divergent	NN	O	O
haplotypes	NN	O	O
suggested	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
,	NN	O	O
recurrent	NN	O	O
N370S	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
different	NN	O	O
conserved	NN	O	O
haplotype	NN	O	O
at	NN	O	O
these	NN	O	O
markers	NN	O	O
was	NN	O	O
identified	NN	O	O
on	NN	O	O
the	NN	O	O
84GG	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
unique	NN	O	O
to	NN	O	O
the	NN	O	O
AJ	NN	O	O
population	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
(	NN	O	O
LD	NN	O	O
)	NN	O	O
delta	NN	O	O
values	NN	O	O
,	NN	O	O
the	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
European	NN	O	O
N370S	NN	O	O
chromosomes	NN	O	O
had	NN	O	O
greater	NN	O	O
haplotype	NN	O	O
diversity	NN	O	O
and	NN	O	O
less	NN	O	O
LD	NN	O	O
at	NN	O	O
the	NN	O	O
markers	NN	O	O
flanking	NN	O	O
the	NN	O	O
conserved	NN	O	O
haplotype	NN	O	O
than	NN	O	O
did	NN	O	O
the	NN	O	O
AJ	NN	O	O
N370S	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
N370S	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
European	NN	O	O
population	NN	O	O
prior	NN	O	O
to	NN	O	O
the	NN	O	O
founding	NN	O	O
of	NN	O	O
the	NN	O	O
AJ	NN	O	O
population	NN	O	O
.	NN	O	O

Coalescence	NN	O	O
analyses	NN	O	O
for	NN	O	O
the	NN	O	O
N370S	NN	O	O
and	NN	O	O
84GG	NN	O	O
mutations	NN	O	O
estimated	NN	O	O
similar	NN	O	O
coalescence	NN	O	O
times	NN	O	O
,	NN	O	O
of	NN	O	O
48	NN	O	O
and	NN	O	O
55	NN	O	O
.	NN	O	O
5	NN	O	O
generations	NN	O	O
ago	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
these	NN	O	O
studies	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
significant	NN	O	O
bottleneck	NN	O	O
occurring	NN	O	O
in	NN	O	O
the	NN	O	O
AJ	NN	O	O
population	NN	O	O
during	NN	O	O
the	NN	O	O
first	NN	O	O
millennium	NN	O	O
,	NN	O	O
when	NN	O	O
the	NN	O	O
population	NN	O	O
became	NN	O	O
established	NN	O	O
in	NN	O	O
Europe	NN	O	O
.	NN	O	O

HLA	NN	O	O
B27	NN	O	O
and	NN	O	O
the	NN	O	O
genetics	NN	O	O
of	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
.	NN	O	O

One	NN	O	O
hundred	NN	O	O
and	NN	O	O
twenty	NN	O	O
-	NN	O	O
eight	NN	O	O
of	NN	O	O
145	NN	O	O
patients	NN	O	O
with	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
HLA	NN	O	O
B27	NN	O	O
positive	NN	O	O
.	NN	O	O

Five	NN	O	O
patients	NN	O	O
had	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
sero	NN	O	O
-	NN	O	O
negative	NN	O	O
peripheral	NN	O	B-Disease
arthritis	NN	O	I-Disease
resembling	NN	O	O
peripheral	NN	O	B-Disease
psoriatic	NN	O	I-Disease
arthritis	NN	O	I-Disease
and	NN	O	O
3	NN	O	O
of	NN	O	O
these	NN	O	O
were	NN	O	O
B27	NN	O	O
negative	NN	O	O
.	NN	O	O

One	NN	O	O
further	NN	O	O
B27	NN	O	O
negative	NN	O	O
patients	NN	O	O
had	NN	O	O
a	NN	O	O
sister	NN	O	O
with	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
and	NN	O	O
ulcerative	NN	O	B-Disease
colitis	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
mother	NN	O	O
with	NN	O	O
ulcerative	NN	O	B-Disease
colitis	NN	O	I-Disease
.	NN	O	O

There	NN	O	O
was	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
somewhat	NN	O	O
later	NN	O	O
age	NN	O	O
of	NN	O	O
onset	NN	O	O
of	NN	O	O
symptoms	NN	O	O
in	NN	O	O
B27	NN	O	O
negative	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
interpreted	NN	O	O
as	NN	O	O
suggesting	NN	O	O
some	NN	O	O
degree	NN	O	O
of	NN	O	O
clinical	NN	O	O
and	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
with	NN	O	O
genes	NN	O	O
for	NN	O	O
psoriasis	NN	O	B-Disease
and	NN	O	O
inflammatory	NN	O	B-Disease
bowel	NN	O	I-Disease
disease	NN	O	I-Disease
being	NN	O	O
important	NN	O	O
in	NN	O	O
some	NN	O	O
individuals	NN	O	O
,	NN	O	O
particularly	NN	O	O
those	NN	O	O
who	NN	O	O
are	NN	O	O
B27	NN	O	O
negative	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
five	NN	O	O
first	NN	O	O
-	NN	O	O
degree	NN	O	O
relatives	NN	O	O
with	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
were	NN	O	O
all	NN	O	O
B27	NN	O	O
positive	NN	O	O
.	NN	O	O

The	NN	O	O
only	NN	O	O
instance	NN	O	O
of	NN	O	O
disassociation	NN	O	O
of	NN	O	O
B27	NN	O	O
and	NN	O	O
spondylitis	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
family	NN	O	O
was	NN	O	O
where	NN	O	O
the	NN	O	O
proband	NN	O	O
had	NN	O	O
ulcerative	NN	O	B-Disease
colitis	NN	O	I-Disease
as	NN	O	O
well	NN	O	O
as	NN	O	O
spondylitis	NN	O	B-Disease
.	NN	O	O

Of	NN	O	O
13	NN	O	O
B27	NN	O	O
positive	NN	O	O
fathers	NN	O	O
3	NN	O	O
could	NN	O	O
be	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
definite	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
(	NN	O	O
23	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
provide	NN	O	O
evidence	NN	O	O
against	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
is	NN	O	O
not	NN	O	O
B27	NN	O	O
but	NN	O	O
a	NN	O	O
closely	NN	O	O
linked	NN	O	O
gene	NN	O	O
and	NN	O	O
favour	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
an	NN	O	O
environmental	NN	O	O
event	NN	O	O
affecting	NN	O	O
approximately	NN	O	O
one	NN	O	O
-	NN	O	O
fifth	NN	O	O
of	NN	O	O
B27	NN	O	O
positive	NN	O	O
males	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
disease	NN	O	O
.	NN	O	O
.	NN	O	O

Founder	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
in	NN	O	O
Polish	NN	O	O
families	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
undertaken	NN	O	O
a	NN	O	O
hospital	NN	O	O
-	NN	O	O
based	NN	O	O
study	NN	O	O
,	NN	O	O
to	NN	O	O
identify	NN	O	O
possible	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
BRCA2	NN	O	O
founder	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Polish	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
group	NN	O	O
consisted	NN	O	O
of	NN	O	O
66	NN	O	O
Polish	NN	O	O
families	NN	O	O
with	NN	O	O
cancer	NN	O	B-Disease
who	NN	O	O
have	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
related	NN	O	O
females	NN	O	O
affected	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
who	NN	O	O
had	NN	O	O
cancer	NN	O	B-Disease
diagnosed	NN	O	O
,	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
affected	NN	O	O
females	NN	O	O
,	NN	O	O
at	NN	O	O
age	NN	O	O
<	NN	O	O
50	NN	O	O
years	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
26	NN	O	O
families	NN	O	O
had	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
,	NN	O	O
4	NN	O	O
families	NN	O	O
had	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
only	NN	O	O
,	NN	O	O
and	NN	O	O
36	NN	O	O
families	NN	O	O
had	NN	O	O
breast	NN	O	B-Disease
cancers	NN	O	I-Disease
only	NN	O	O
.	NN	O	O

Genomic	NN	O	O
DNA	NN	O	O
was	NN	O	O
prepared	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
affected	NN	O	O
woman	NN	O	O
from	NN	O	O
each	NN	O	O
family	NN	O	O
.	NN	O	O

The	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
BRCA2	NN	O	O
was	NN	O	O
screened	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
,	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
SSCP	NN	O	O
followed	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
observed	NN	O	O
variants	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
35	NN	O	O
(	NN	O	O
53	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
66	NN	O	O
families	NN	O	O
studied	NN	O	O
.	NN	O	O

All	NN	O	O
but	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
within	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
.	NN	O	O

BRCA1	NN	O	B-Disease
abnormalities	NN	O	I-Disease
were	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
four	NN	O	O
families	NN	O	O
with	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
only	NN	O	O
,	NN	O	O
in	NN	O	O
67	NN	O	O
%	NN	O	O
of	NN	O	O
27	NN	O	O
families	NN	O	O
with	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
in	NN	O	O
34	NN	O	O
%	NN	O	O
of	NN	O	O
35	NN	O	O
families	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
only	NN	O	O
.	NN	O	O

The	NN	O	O
single	NN	O	O
family	NN	O	O
with	NN	O	O
a	NN	O	O
BRCA2	NN	O	O
mutation	NN	O	O
had	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Seven	NN	O	O
distinct	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
;	NN	O	O
five	NN	O	O
of	NN	O	O
these	NN	O	O
occurred	NN	O	O
in	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
total	NN	O	O
,	NN	O	O
recurrent	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
33	NN	O	O
(	NN	O	O
94	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
35	NN	O	O
families	NN	O	O
with	NN	O	O
detected	NN	O	O
mutations	NN	O	O
.	NN	O	O

Three	NN	O	O
BRCA1	NN	O	B-Disease
abnormalities	NN	O	I-Disease
-	NN	O	O
5382insC	NN	O	O
,	NN	O	O
C61G	NN	O	O
,	NN	O	O
and	NN	O	O
4153delA	NN	O	O
-	NN	O	O
accounted	NN	O	O
for	NN	O	O
51	NN	O	O
%	NN	O	O
,	NN	O	O
20	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
11	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
identified	NN	O	O
mutations	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
very	NN	O	B-Disease
long	NN	O	I-Disease
chain	NN	O	I-Disease
acyl	NN	O	I-Disease
-	NN	O	I-Disease
CoA	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
three	NN	O	O
Israeli	NN	O	O
patients	NN	O	O
:	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
mutant	NN	O	O
allele	NN	O	O
with	NN	O	O
P65L	NN	O	O
and	NN	O	O
K247Q	NN	O	O
mutations	NN	O	O
,	NN	O	O
the	NN	O	O
former	NN	O	O
being	NN	O	O
an	NN	O	O
exonic	NN	O	O
mutation	NN	O	O
causing	NN	O	O
exon	NN	O	O
3	NN	O	O
skipping	NN	O	O
.	NN	O	O

Very	NN	O	B-Disease
long	NN	O	I-Disease
chain	NN	O	I-Disease
acyl	NN	O	I-Disease
-	NN	O	I-Disease
CoA	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
VLCAD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
life	NN	O	O
-	NN	O	O
threatening	NN	O	O
disorder	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
fatty	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
four	NN	O	O
novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
three	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
had	NN	O	O
the	NN	O	O
severe	NN	O	O
childhood	NN	O	O
form	NN	O	O
of	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
early	NN	O	O
onset	NN	O	O
and	NN	O	O
high	NN	O	O
mortality	NN	O	O
.	NN	O	O

Immunoblot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
VLCAD	NN	O	O
protein	NN	O	O
was	NN	O	O
undetectable	NN	O	O
in	NN	O	O
patients	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
,	NN	O	O
whereas	NN	O	O
normal	NN	O	O
-	NN	O	O
size	NN	O	O
VLCAD	NN	O	O
protein	NN	O	O
and	NN	O	O
an	NN	O	O
aberrant	NN	O	O
form	NN	O	O
of	NN	O	O
VLCAD	NN	O	O
(	NN	O	O
4kDa	NN	O	O
smaller	NN	O	O
)	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
patient	NN	O	O
1	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
null	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
patient	NN	O	O
2	NN	O	O
is	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
,	NN	O	O
del	NN	O	O
4	NN	O	O
bp	NN	O	O
at	NN	O	O
798	NN	O	O
-	NN	O	O
801	NN	O	O
,	NN	O	O
and	NN	O	O
patient	NN	O	O
3	NN	O	O
is	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
65C	NN	O	O
>	NN	O	O
A	NN	O	O
(	NN	O	O
S22X	NN	O	O
)	NN	O	O
.	NN	O	O

Patient	NN	O	O
1	NN	O	O
was	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
complex	NN	O	O
mutant	NN	O	O
allele	NN	O	O
containing	NN	O	O
two	NN	O	O
alterations	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
194C	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
(	NN	O	O
P65L	NN	O	O
)	NN	O	O
and	NN	O	O
739A	NN	O	O
>	NN	O	O
C	NN	O	O
transversion	NN	O	O
(	NN	O	O
K247Q	NN	O	O
)	NN	O	O
;	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
P65L	NN	O	O
,	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
change	NN	O	O
does	NN	O	O
not	NN	O	O
reduce	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
change	NN	O	O
resulted	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
skipping	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
latter	NN	O	O
K247Q	NN	O	O
mutation	NN	O	O
had	NN	O	O
a	NN	O	O
drastic	NN	O	O
effect	NN	O	O
on	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
verified	NN	O	O
these	NN	O	O
events	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
splicing	NN	O	O
experiments	NN	O	O
and	NN	O	O
transient	NN	O	O
expression	NN	O	O
analysis	NN	O	O
of	NN	O	O
mutant	NN	O	O
cDNAs	NN	O	O
.	NN	O	O

The	NN	O	O
P65L	NN	O	O
mutation	NN	O	O
locates	NN	O	O
11	NN	O	O
bases	NN	O	O
upstream	NN	O	O
of	NN	O	O
a	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
3	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
an	NN	O	O
example	NN	O	O
of	NN	O	O
an	NN	O	O
exonic	NN	O	O
mutation	NN	O	O
which	NN	O	O
affects	NN	O	O
exon	NN	O	O
-	NN	O	O
splicing	NN	O	O
.	NN	O	O
.	NN	O	O

Submicroscopic	NN	O	O
deletion	NN	O	O
in	NN	O	O
cousins	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
causes	NN	O	O
a	NN	O	O
grandmatrilineal	NN	O	O
inheritance	NN	O	O
pattern	NN	O	O
:	NN	O	O
effects	NN	O	O
of	NN	O	O
imprinting	NN	O	O
.	NN	O	O

The	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
critical	NN	O	O
region	NN	O	O
on	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
is	NN	O	O
subject	NN	O	O
to	NN	O	O
imprinting	NN	O	O
.	NN	O	O

PWS	NN	O	B-Disease
becomes	NN	O	O
apparent	NN	O	O
when	NN	O	O
genes	NN	O	O
on	NN	O	O
the	NN	O	O
paternally	NN	O	O
inherited	NN	O	O
chromosome	NN	O	O
are	NN	O	O
not	NN	O	O
expressed	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
PWS	NN	O	I-Disease
is	NN	O	O
rare	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
a	NN	O	O
male	NN	O	O
and	NN	O	O
a	NN	O	O
female	NN	O	O
paternal	NN	O	O
first	NN	O	O
cousin	NN	O	O
both	NN	O	O
have	NN	O	O
PWS	NN	O	B-Disease
with	NN	O	O
cytogenetically	NN	O	O
normal	NN	O	O
karyotypes	NN	O	O
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
analysis	NN	O	O
shows	NN	O	O
a	NN	O	O
submicroscopic	NN	O	O
deletion	NN	O	O
of	NN	O	O
SNRPN	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
closely	NN	O	O
associated	NN	O	O
loci	NN	O	O
D15S10	NN	O	O
,	NN	O	O
D15S11	NN	O	O
,	NN	O	O
D15S63	NN	O	O
,	NN	O	O
and	NN	O	O
GABRB3	NN	O	O
.	NN	O	O

The	NN	O	O
cousins	NN	O	O
fathers	NN	O	O
and	NN	O	O
two	NN	O	O
paternal	NN	O	O
aunts	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
deletion	NN	O	O
and	NN	O	O
are	NN	O	O
clinically	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
grandmother	NN	O	O
of	NN	O	O
the	NN	O	O
cousins	NN	O	O
is	NN	O	O
deceased	NN	O	O
and	NN	O	O
not	NN	O	O
available	NN	O	O
for	NN	O	O
study	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
grandfather	NN	O	O
is	NN	O	O
not	NN	O	O
deleted	NN	O	O
for	NN	O	O
SNRPN	NN	O	O
.	NN	O	O

DNA	NN	O	O
methylation	NN	O	O
analysis	NN	O	O
of	NN	O	O
D15S63	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
an	NN	O	O
abnormality	NN	O	O
of	NN	O	O
the	NN	O	O
imprinting	NN	O	O
center	NN	O	O
associated	NN	O	O
with	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

"	NN	O	O
Grandmatrilineal	NN	O	O
"	NN	O	O
inheritance	NN	O	O
occurs	NN	O	O
when	NN	O	O
a	NN	O	O
woman	NN	O	O
with	NN	O	O
deletion	NN	O	O
of	NN	O	O
an	NN	O	O
imprinted	NN	O	O
,	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
gene	NN	O	O
is	NN	O	O
at	NN	O	O
risk	NN	O	O
of	NN	O	O
having	NN	O	O
affected	NN	O	O
grandchildren	NN	O	O
through	NN	O	O
her	NN	O	O
sons	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
case	NN	O	O
,	NN	O	O
PWS	NN	O	B-Disease
does	NN	O	O
not	NN	O	O
become	NN	O	O
evident	NN	O	O
as	NN	O	O
long	NN	O	O
as	NN	O	O
the	NN	O	O
deletion	NN	O	O
is	NN	O	O
passed	NN	O	O
through	NN	O	O
the	NN	O	O
matrilineal	NN	O	O
line	NN	O	O
.	NN	O	O

This	NN	O	O
represents	NN	O	O
a	NN	O	O
unique	NN	O	O
inheritance	NN	O	O
pattern	NN	O	O
due	NN	O	O
to	NN	O	O
imprinting	NN	O	O
.	NN	O	O
.	NN	O	O

Human	NN	O	O
glycine	NN	O	O
decarboxylase	NN	O	O
gene	NN	O	O
(	NN	O	O
GLDC	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
highly	NN	O	O
conserved	NN	O	O
processed	NN	O	O
pseudogene	NN	O	O
(	NN	O	O
psiGLDC	NN	O	O
)	NN	O	O
:	NN	O	O
their	NN	O	O
structure	NN	O	O
and	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
deletion	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
nonketotic	NN	O	B-Disease
hyperglycinemia	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
glycine	NN	O	O
decarboxylase	NN	O	O
gene	NN	O	O
(	NN	O	O
GLDC	NN	O	O
)	NN	O	O
cause	NN	O	O
nonketotic	NN	O	B-Disease
hyperglycinemia	NN	O	I-Disease
(	NN	O	O
NKH	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
an	NN	O	O
in	NN	O	B-Disease
-	NN	O	I-Disease
born	NN	O	I-Disease
error	NN	O	I-Disease
of	NN	O	I-Disease
metabolism	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
severe	NN	O	O
neurological	NN	O	B-Disease
disturbance	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
GLDC	NN	O	O
and	NN	O	O
of	NN	O	O
its	NN	O	O
pseudogene	NN	O	O
(	NN	O	O
psiGLDC	NN	O	O
)	NN	O	O
and	NN	O	O
studied	NN	O	O
their	NN	O	O
expression	NN	O	O
for	NN	O	O
a	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
NKH	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
GLDC	NN	O	O
gene	NN	O	O
spans	NN	O	O
at	NN	O	O
least	NN	O	O
135	NN	O	O
kb	NN	O	O
and	NN	O	O
consists	NN	O	O
of	NN	O	O
25	NN	O	O
exons	NN	O	O
.	NN	O	O

All	NN	O	O
donor	NN	O	O
and	NN	O	O
acceptor	NN	O	O
sites	NN	O	O
adhere	NN	O	O
to	NN	O	O
the	NN	O	O
canonical	NN	O	O
GT	NN	O	O
-	NN	O	O
AG	NN	O	O
rule	NN	O	O
,	NN	O	O
except	NN	O	O
for	NN	O	O
the	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
21	NN	O	O
,	NN	O	O
where	NN	O	O
a	NN	O	O
variant	NN	O	O
form	NN	O	O
GC	NN	O	O
is	NN	O	O
used	NN	O	O
instead	NN	O	O
of	NN	O	O
GT	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
site	NN	O	O
has	NN	O	O
been	NN	O	O
assigned	NN	O	O
to	NN	O	O
a	NN	O	O
residue	NN	O	O
163	NN	O	O
bp	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
translation	NN	O	O
initiation	NN	O	O
triplet	NN	O	O
by	NN	O	O
primer	NN	O	O
extension	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
psiGLDC	NN	O	O
gene	NN	O	O
has	NN	O	O
no	NN	O	O
intron	NN	O	O
and	NN	O	O
shares	NN	O	O
97	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
functional	NN	O	O
GLDC	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
psiGLDC	NN	O	O
is	NN	O	O
a	NN	O	O
processed	NN	O	O
pseudogene	NN	O	O
that	NN	O	O
arose	NN	O	O
from	NN	O	O
the	NN	O	O
GLDC	NN	O	O
transcript	NN	O	O
about	NN	O	O
4	NN	O	O
-	NN	O	O
8	NN	O	O
million	NN	O	O
years	NN	O	O
ago	NN	O	O
.	NN	O	O

RNA	NN	O	O
blotting	NN	O	O
analysis	NN	O	O
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
GLDC	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	O
liver	NN	O	O
,	NN	O	O
kidney	NN	O	O
,	NN	O	O
brain	NN	O	O
,	NN	O	O
and	NN	O	O
placenta	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
examined	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
NKH	NN	O	B-Disease
with	NN	O	O
no	NN	O	O
detectable	NN	O	O
GLDC	NN	O	O
mRNA	NN	O	O
in	NN	O	O
his	NN	O	O
lymphoblasts	NN	O	O
.	NN	O	O

Exons	NN	O	O
1	NN	O	O
-	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
GLDC	NN	O	O
gene	NN	O	O
from	NN	O	O
this	NN	O	O
patient	NN	O	O
are	NN	O	O
not	NN	O	O
amplified	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
those	NN	O	O
from	NN	O	O
control	NN	O	O
subjects	NN	O	O
are	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
large	NN	O	O
homozygous	NN	O	O
deletion	NN	O	O
(	NN	O	O
at	NN	O	O
least	NN	O	O
30	NN	O	O
kb	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
devised	NN	O	O
a	NN	O	O
semi	NN	O	O
-	NN	O	O
quantitative	NN	O	O
PCR	NN	O	O
to	NN	O	O
estimate	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GLDC	NN	O	O
alleles	NN	O	O
by	NN	O	O
using	NN	O	O
psiGLDC	NN	O	O
as	NN	O	O
an	NN	O	O
internal	NN	O	O
control	NN	O	O
and	NN	O	O
have	NN	O	O
confirmed	NN	O	O
the	NN	O	O
homozygosity	NN	O	O
and	NN	O	O
heterozygosity	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
his	NN	O	O
parents	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Structural	NN	O	O
information	NN	O	O
of	NN	O	O
GLDC	NN	O	O
and	NN	O	O
psiGLDC	NN	O	O
should	NN	O	O
facilitate	NN	O	O
the	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
NKH	NN	O	B-Disease
.	NN	O	O

De	NN	O	O
novo	NN	O	O
deletions	NN	O	O
of	NN	O	O
SNRPN	NN	O	O
exon	NN	O	O
1	NN	O	O
in	NN	O	O
early	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
embryos	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
paternal	NN	O	O
to	NN	O	O
maternal	NN	O	O
imprint	NN	O	O
switch	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
neurogenetic	NN	O	B-Disease
disease	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
infantile	NN	O	B-Disease
hypotonia	NN	O	I-Disease
,	NN	O	O
gonadal	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
obsessive	NN	O	O
behaviour	NN	O	O
and	NN	O	O
neonatal	NN	O	O
feeding	NN	O	O
difficulties	NN	O	O
followed	NN	O	O
by	NN	O	O
hyperphagia	NN	O	B-Disease
,	NN	O	O
leading	NN	O	O
to	NN	O	O
profound	NN	O	O
obesity	NN	O	B-Disease
.	NN	O	O

PWS	NN	O	B-Disease
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
paternal	NN	O	O
genetic	NN	O	O
information	NN	O	O
at	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

Five	NN	O	O
imprinted	NN	O	O
,	NN	O	O
paternally	NN	O	O
expressed	NN	O	O
genes	NN	O	O
map	NN	O	O
to	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
region	NN	O	O
,	NN	O	O
MKRN3	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
NDN	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
NDNL1	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
5	NN	O	O
)	NN	O	O
,	NN	O	O
SNRPN	NN	O	O
(	NN	O	O
refs	NN	O	O
6	NN	O	O
-	NN	O	O
8	NN	O	O
)	NN	O	O
and	NN	O	O
IPW	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
9	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
two	NN	O	O
poorly	NN	O	O
characterized	NN	O	O
framents	NN	O	O
designated	NN	O	O
PAR	NN	O	O
-	NN	O	O
1	NN	O	O
and	NN	O	O
PAR	NN	O	O
-	NN	O	O
5	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
10	NN	O	O
)	NN	O	O
.	NN	O	O

Imprinting	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
involves	NN	O	O
a	NN	O	O
bipartite	NN	O	O
imprinting	NN	O	O
centre	NN	O	O
(	NN	O	O
IC	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
overlaps	NN	O	O
SNRPN	NN	O	O
(	NN	O	O
refs	NN	O	O
10	NN	O	O
,	NN	O	O
11	NN	O	O
)	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
SNRPN	NN	O	O
promoter	NN	O	O
/	NN	O	O
exon	NN	O	O
1	NN	O	O
region	NN	O	O
(	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
IC	NN	O	O
element	NN	O	O
)	NN	O	O
appears	NN	O	O
to	NN	O	O
impair	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
imprint	NN	O	O
in	NN	O	O
the	NN	O	O
male	NN	O	O
germ	NN	O	O
line	NN	O	O
and	NN	O	O
leads	NN	O	O
to	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
a	NN	O	O
PWS	NN	O	B-Disease
family	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
father	NN	O	O
is	NN	O	O
mosaic	NN	O	O
for	NN	O	O
an	NN	O	O
IC	NN	O	O
deletion	NN	O	O
on	NN	O	O
his	NN	O	O
paternal	NN	O	O
chromosome	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
chromosome	NN	O	O
has	NN	O	O
acquired	NN	O	O
a	NN	O	O
maternal	NN	O	O
methylation	NN	O	O
imprint	NN	O	O
in	NN	O	O
his	NN	O	O
somatic	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
made	NN	O	O
identical	NN	O	O
findings	NN	O	O
in	NN	O	O
chimaeric	NN	O	O
mice	NN	O	O
generated	NN	O	O
from	NN	O	O
two	NN	O	O
independent	NN	O	O
embryonic	NN	O	O
stem	NN	O	O
(	NN	O	O
ES	NN	O	O
)	NN	O	O
cell	NN	O	O
lines	NN	O	O
harbouring	NN	O	O
a	NN	O	O
similar	NN	O	O
deletion	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
IC	NN	O	O
element	NN	O	O
is	NN	O	O
not	NN	O	O
only	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
imprint	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
for	NN	O	O
its	NN	O	O
postzygotic	NN	O	O
maintenance	NN	O	O
.	NN	O	O
.	NN	O	O

Mice	NN	O	O
deficient	NN	O	B-Disease
in	NN	O	I-Disease
Six5	NN	O	I-Disease
develop	NN	O	O
cataracts	NN	O	B-Disease
:	NN	O	O
implications	NN	O	O
for	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Expansion	NN	O	O
of	NN	O	O
a	NN	O	O
CTG	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
UTR	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
DMPK	NN	O	O
at	NN	O	O
the	NN	O	O
DM1	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
19	NN	O	O
causes	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
dominantly	NN	O	B-Disease
inherited	NN	O	I-Disease
disease	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
skeletal	NN	O	O
muscle	NN	O	B-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
myotonia	NN	O	B-Disease
,	NN	O	O
cataracts	NN	O	B-Disease
and	NN	O	O
cardiac	NN	O	B-Disease
conduction	NN	O	I-Disease
defects	NN	O	I-Disease
.	NN	O	O

Targeted	NN	O	O
deletion	NN	O	O
of	NN	O	O
Dm15	NN	O	O
,	NN	O	O
the	NN	O	O
mouse	NN	O	O
orthologue	NN	O	O
of	NN	O	O
human	NN	O	O
DMPK	NN	O	O
,	NN	O	O
produced	NN	O	O
mice	NN	O	O
with	NN	O	O
a	NN	O	O
mild	NN	O	O
myopathy	NN	O	B-Disease
and	NN	O	O
cardiac	NN	O	B-Disease
conduction	NN	O	I-Disease
abnormalities	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
without	NN	O	O
other	NN	O	O
features	NN	O	O
of	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
such	NN	O	O
as	NN	O	O
myotonia	NN	O	B-Disease
and	NN	O	O
cataracts	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
,	NN	O	O
and	NN	O	O
others	NN	O	O
,	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
decreases	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
adjacent	NN	O	O
gene	NN	O	O
SIX5	NN	O	O
(	NN	O	O
refs	NN	O	O
7	NN	O	O
,	NN	O	O
8	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
homeodomain	NN	O	O
transcription	NN	O	O
factor	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
SIX5	NN	O	B-Disease
deficiency	NN	O	I-Disease
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
phenotype	NN	O	O
,	NN	O	O
we	NN	O	O
disrupted	NN	O	O
mouse	NN	O	O
Six5	NN	O	O
by	NN	O	O
replacing	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	O
with	NN	O	O
a	NN	O	O
beta	NN	O	O
-	NN	O	O
galactosidase	NN	O	O
reporter	NN	O	O
.	NN	O	O

Six5	NN	O	O
-	NN	O	O
mutant	NN	O	O
mice	NN	O	O
showed	NN	O	O
reporter	NN	O	O
expression	NN	O	O
in	NN	O	O
multiple	NN	O	O
tissues	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
developing	NN	O	O
lens	NN	O	O
.	NN	O	O

Homozygous	NN	O	O
mutant	NN	O	O
mice	NN	O	O
had	NN	O	O
no	NN	O	O
apparent	NN	O	O
abnormalities	NN	O	B-Disease
of	NN	O	I-Disease
skeletal	NN	O	I-Disease
muscle	NN	O	I-Disease
function	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
developed	NN	O	O
lenticular	NN	O	B-Disease
opacities	NN	O	I-Disease
at	NN	O	O
a	NN	O	O
higher	NN	O	O
rate	NN	O	O
than	NN	O	O
controls	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
SIX5	NN	O	B-Disease
deficiency	NN	O	I-Disease
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
cataract	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
that	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
represents	NN	O	O
a	NN	O	O
multigenic	NN	O	B-Disease
disorder	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Heterozygous	NN	O	O
loss	NN	O	O
of	NN	O	O
Six5	NN	O	O
in	NN	O	O
mice	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
cause	NN	O	O
ocular	NN	O	O
cataracts	NN	O	B-Disease
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
skeletal	NN	O	O
muscle	NN	O	B-Disease
wasting	NN	O	I-Disease
,	NN	O	O
myotonia	NN	O	B-Disease
,	NN	O	O
cardiac	NN	O	B-Disease
arrhythmia	NN	O	I-Disease
,	NN	O	O
hyperinsulinaemia	NN	O	B-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
and	NN	O	O
ocular	NN	O	O
cataracts	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
in	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
DMPK	NN	O	O
and	NN	O	O
5	NN	O	O
of	NN	O	O
a	NN	O	O
homeodomain	NN	O	O
-	NN	O	O
encoding	NN	O	O
gene	NN	O	O
,	NN	O	O
SIX5	NN	O	O
(	NN	O	O
formerly	NN	O	O
DMAHP	NN	O	O
;	NN	O	O
refs	NN	O	O
2	NN	O	O
-	NN	O	O
5	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
three	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

First	NN	O	O
,	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
may	NN	O	O
alter	NN	O	O
the	NN	O	O
processing	NN	O	O
or	NN	O	O
transport	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
DMPK	NN	O	O
mRNA	NN	O	O
and	NN	O	O
consequently	NN	O	O
reduce	NN	O	O
DMPK	NN	O	O
levels	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
may	NN	O	O
establish	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
heterochromatin	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
sequence	NN	O	O
and	NN	O	O
decrease	NN	O	O
SIX5	NN	O	O
transcription	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
toxic	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
may	NN	O	O
be	NN	O	O
intrinsic	NN	O	O
to	NN	O	O
the	NN	O	O
repeated	NN	O	O
elements	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
DNA	NN	O	O
or	NN	O	O
RNA	NN	O	O
(	NN	O	O
refs	NN	O	O
10	NN	O	O
,	NN	O	O
11	NN	O	O
)	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
dose	NN	O	O
-	NN	O	O
dependent	NN	O	O
loss	NN	O	O
of	NN	O	O
Dm15	NN	O	O
(	NN	O	O
the	NN	O	O
mouse	NN	O	O
DMPK	NN	O	O
homologue	NN	O	O
)	NN	O	O
in	NN	O	O
mice	NN	O	O
produces	NN	O	O
a	NN	O	O
partial	NN	O	O
DM	NN	O	B-Disease
phenotype	NN	O	O
characterized	NN	O	O
by	NN	O	O
decreased	NN	O	O
development	NN	O	O
of	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
force	NN	O	O
and	NN	O	O
cardiac	NN	O	B-Disease
conduction	NN	O	I-Disease
disorders	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
test	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Six5	NN	O	O
loss	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
a	NN	O	O
strain	NN	O	O
of	NN	O	O
mice	NN	O	O
in	NN	O	O
which	NN	O	O
Six5	NN	O	O
was	NN	O	O
deleted	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
rate	NN	O	O
and	NN	O	O
severity	NN	O	O
of	NN	O	O
cataract	NN	O	B-Disease
formation	NN	O	O
is	NN	O	O
inversely	NN	O	O
related	NN	O	O
to	NN	O	O
Six5	NN	O	O
dosage	NN	O	O
and	NN	O	O
is	NN	O	O
temporally	NN	O	O
progressive	NN	O	O
.	NN	O	O

Six5	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
and	NN	O	O
Six5	NN	O	O
-	NN	O	O
/	NN	O	O
-	NN	O	O
mice	NN	O	O
show	NN	O	O
increased	NN	O	O
steady	NN	O	O
-	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
K	NN	O	O
+	NN	O	O
-	NN	O	O
ATPase	NN	O	O
alpha	NN	O	O
-	NN	O	O
1	NN	O	O
subunit	NN	O	O
and	NN	O	O
decreased	NN	O	O
Dm15	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
altered	NN	O	O
ion	NN	O	O
homeostasis	NN	O	O
within	NN	O	O
the	NN	O	O
lens	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
cataract	NN	O	B-Disease
formation	NN	O	O
.	NN	O	O

As	NN	O	O
ocular	NN	O	O
cataracts	NN	O	B-Disease
are	NN	O	O
a	NN	O	O
characteristic	NN	O	O
feature	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
decreased	NN	O	O
SIX5	NN	O	O
transcription	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
aetiology	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
contiguous	NN	O	O
gene	NN	O	O
syndrome	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
partial	NN	O	O
loss	NN	O	O
of	NN	O	O
both	NN	O	O
DMPK	NN	O	O
and	NN	O	O
SIX5	NN	O	O
.	NN	O	O
.	NN	O	O

ATM	NN	O	O
-	NN	O	O
dependent	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
nibrin	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
radiation	NN	O	O
exposure	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
ATM	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
disorder	NN	O	I-Disease
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
cerebellar	NN	O	B-Disease
dysfunction	NN	O	I-Disease
,	NN	O	O
radiosensitivity	NN	O	O
,	NN	O	O
chromosomal	NN	O	O
instability	NN	O	O
and	NN	O	O
cancer	NN	O	B-Disease
predisposition	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
phenotype	NN	O	O
and	NN	O	O
the	NN	O	O
similarity	NN	O	O
of	NN	O	O
the	NN	O	O
ATM	NN	O	O
protein	NN	O	O
to	NN	O	O
other	NN	O	O
DNA	NN	O	O
-	NN	O	O
damage	NN	O	O
sensors	NN	O	O
suggests	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
ATM	NN	O	O
in	NN	O	O
biochemical	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
recognition	NN	O	O
,	NN	O	O
signalling	NN	O	O
and	NN	O	O
repair	NN	O	O
of	NN	O	O
DNA	NN	O	O
double	NN	O	O
-	NN	O	O
strand	NN	O	O
breaks	NN	O	O
(	NN	O	O
DSBs	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
strong	NN	O	O
parallels	NN	O	O
between	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
radiosensitivity	NN	O	O
,	NN	O	O
chromosomal	NN	O	O
instability	NN	O	O
and	NN	O	O
cancer	NN	O	B-Disease
predisposition	NN	O	O
in	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
and	NN	O	O
that	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Nijmegen	NN	O	B-Disease
breakage	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
NBS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
defective	NN	O	O
in	NN	O	O
NBS	NN	O	B-Disease
,	NN	O	O
nibrin	NN	O	O
(	NN	O	O
encoded	NN	O	O
by	NN	O	O
NBS1	NN	O	O
)	NN	O	O
,	NN	O	O
forms	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
MRE11	NN	O	O
and	NN	O	O
RAD50	NN	O	O
(	NN	O	O
refs	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
complex	NN	O	O
localizes	NN	O	O
to	NN	O	O
DSBs	NN	O	O
within	NN	O	O
30	NN	O	O
minutes	NN	O	O
after	NN	O	O
cellular	NN	O	O
exposure	NN	O	O
to	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
(	NN	O	O
IR	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
observed	NN	O	O
in	NN	O	O
brightly	NN	O	O
staining	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
after	NN	O	O
a	NN	O	O
longer	NN	O	O
period	NN	O	O
of	NN	O	O
time	NN	O	O
.	NN	O	O

The	NN	O	O
overlap	NN	O	O
between	NN	O	O
clinical	NN	O	O
and	NN	O	O
cellular	NN	O	O
phenotypes	NN	O	O
in	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
and	NN	O	O
NBS	NN	O	B-Disease
suggests	NN	O	O
that	NN	O	O
ATM	NN	O	O
and	NN	O	O
nibrin	NN	O	O
may	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
biochemical	NN	O	O
pathway	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
nibrin	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
within	NN	O	O
one	NN	O	O
hour	NN	O	O
of	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
IR	NN	O	O
.	NN	O	O

This	NN	O	O
response	NN	O	O
is	NN	O	O
abrogated	NN	O	O
in	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
cells	NN	O	O
that	NN	O	O
either	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
ATM	NN	O	O
protein	NN	O	O
or	NN	O	O
express	NN	O	O
near	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
mutant	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
ATM	NN	O	O
physically	NN	O	O
interacts	NN	O	O
with	NN	O	O
and	NN	O	O
phosphorylates	NN	O	O
nibrin	NN	O	O
on	NN	O	O
serine	NN	O	O
343	NN	O	O
both	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
functionally	NN	O	O
important	NN	O	O
because	NN	O	O
mutated	NN	O	O
nibrin	NN	O	O
(	NN	O	O
S343A	NN	O	O
)	NN	O	O
does	NN	O	O
not	NN	O	O
completely	NN	O	O
complement	NN	O	O
radiosensitivity	NN	O	O
in	NN	O	O
NBS	NN	O	B-Disease
cells	NN	O	O
.	NN	O	O

ATM	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
nibrin	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
nibrin	NN	O	O
-	NN	O	O
MRE11	NN	O	O
-	NN	O	O
RAD50	NN	O	O
association	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
radiation	NN	O	O
-	NN	O	O
induced	NN	O	O
foci	NN	O	O
formation	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
a	NN	O	O
biochemical	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
similarity	NN	O	O
in	NN	O	O
phenotype	NN	O	O
between	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
and	NN	O	O
NBS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Clinicopathologic	NN	O	O
features	NN	O	O
of	NN	O	O
BRCA	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
and	NN	O	I-Disease
sporadic	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

CONTEXT	NN	O	O
Most	NN	O	O
hereditary	NN	O	B-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
are	NN	O	O
associated	NN	O	O
with	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
.	NN	O	O

Attempts	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
BRCA	NN	O	O
mutation	NN	O	O
status	NN	O	O
in	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
have	NN	O	O
produced	NN	O	O
conflicting	NN	O	O
results	NN	O	O
,	NN	O	O
especially	NN	O	O
regarding	NN	O	O
survival	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
To	NN	O	O
determine	NN	O	O
whether	NN	O	O
hereditary	NN	O	B-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
have	NN	O	O
distinct	NN	O	O
clinical	NN	O	O
and	NN	O	O
pathological	NN	O	O
features	NN	O	O
compared	NN	O	O
with	NN	O	O
sporadic	NN	O	B-Disease
(	NN	O	I-Disease
nonhereditary	NN	O	I-Disease
)	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

DESIGN	NN	O	O
AND	NN	O	O
SETTING	NN	O	O
Retrospective	NN	O	O
cohort	NN	O	O
study	NN	O	O
of	NN	O	O
a	NN	O	O
consecutive	NN	O	O
series	NN	O	O
of	NN	O	O
933	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
diagnosed	NN	O	O
and	NN	O	O
treated	NN	O	O
at	NN	O	O
our	NN	O	O
institution	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
cancer	NN	O	B-Disease
center	NN	O	O
as	NN	O	O
designated	NN	O	O
by	NN	O	O
the	NN	O	O
National	NN	O	O
Cancer	NN	O	B-Disease
Institute	NN	O	O
,	NN	O	O
over	NN	O	O
a	NN	O	O
12	NN	O	O
-	NN	O	O
year	NN	O	O
period	NN	O	O
(	NN	O	O
December	NN	O	O
1986	NN	O	O
to	NN	O	O
August	NN	O	O
1998	NN	O	O
)	NN	O	O
.	NN	O	O

PATIENTS	NN	O	O
The	NN	O	O
study	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
patients	NN	O	O
of	NN	O	O
Jewish	NN	O	O
origin	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
ease	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
BRCA2	NN	O	O
genotyping	NN	O	O
in	NN	O	O
this	NN	O	O
ethnic	NN	O	O
group	NN	O	O
.	NN	O	O

From	NN	O	O
the	NN	O	O
189	NN	O	O
patients	NN	O	O
who	NN	O	O
identified	NN	O	O
themselves	NN	O	O
as	NN	O	O
Jewish	NN	O	O
,	NN	O	O
88	NN	O	O
hereditary	NN	O	O
cases	NN	O	O
were	NN	O	O
identified	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
germline	NN	O	O
founder	NN	O	O
mutation	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
BRCA2	NN	O	O
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
101	NN	O	O
cases	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
series	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
BRCA	NN	O	O
mutation	NN	O	O
and	NN	O	O
2	NN	O	O
additional	NN	O	O
groups	NN	O	O
(	NN	O	O
Gynecologic	NN	O	O
Oncology	NN	O	O
Group	NN	O	O
protocols	NN	O	O
52	NN	O	O
and	NN	O	O
111	NN	O	O
)	NN	O	O
with	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
from	NN	O	O
clinical	NN	O	O
trials	NN	O	O
(	NN	O	O
for	NN	O	O
the	NN	O	O
survival	NN	O	O
analysis	NN	O	O
)	NN	O	O
were	NN	O	O
included	NN	O	O
for	NN	O	O
comparison	NN	O	O
.	NN	O	O

MAIN	NN	O	O
OUTCOME	NN	O	O
MEASURES	NN	O	O
Age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
surgical	NN	O	O
stage	NN	O	O
,	NN	O	O
histologic	NN	O	O
cell	NN	O	O
type	NN	O	O
and	NN	O	O
grade	NN	O	O
,	NN	O	O
and	NN	O	O
surgical	NN	O	O
outcome	NN	O	O
;	NN	O	O
and	NN	O	O
response	NN	O	O
to	NN	O	O
chemotherapy	NN	O	O
and	NN	O	O
survival	NN	O	O
for	NN	O	O
advanced	NN	O	O
-	NN	O	O
stage	NN	O	O
(	NN	O	O
II	NN	O	O
and	NN	O	O
IV	NN	O	O
)	NN	O	O
cases	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
Hereditary	NN	O	B-Disease
cancers	NN	O	I-Disease
were	NN	O	O
rarely	NN	O	O
diagnosed	NN	O	O
before	NN	O	O
age	NN	O	O
40	NN	O	O
years	NN	O	O
and	NN	O	O
were	NN	O	O
common	NN	O	O
after	NN	O	O
age	NN	O	O
60	NN	O	O
years	NN	O	O
,	NN	O	O
with	NN	O	O
mean	NN	O	O
age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
being	NN	O	O
significantly	NN	O	O
younger	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
vs	NN	O	O
BRCA2	NN	O	O
-	NN	O	O
linked	NN	O	O
patients	NN	O	O
(	NN	O	O
54	NN	O	O
vs	NN	O	O
62	NN	O	O
years	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
04	NN	O	O
)	NN	O	O
.	NN	O	O

Histology	NN	O	O
,	NN	O	O
grade	NN	O	O
,	NN	O	O
stage	NN	O	O
,	NN	O	O
and	NN	O	O
success	NN	O	O
of	NN	O	O
cytoreductive	NN	O	O
surgery	NN	O	O
were	NN	O	O
similar	NN	O	O
for	NN	O	O
hereditary	NN	O	O
and	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
.	NN	O	O

The	NN	O	O
hereditary	NN	O	O
group	NN	O	O
had	NN	O	O
a	NN	O	O
longer	NN	O	O
disease	NN	O	O
-	NN	O	O
free	NN	O	O
interval	NN	O	O
following	NN	O	O
primary	NN	O	O
chemotherapy	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
the	NN	O	O
nonhereditary	NN	O	O
group	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
median	NN	O	O
time	NN	O	O
to	NN	O	O
recurrence	NN	O	O
of	NN	O	O
14	NN	O	O
months	NN	O	O
and	NN	O	O
7	NN	O	O
months	NN	O	O
,	NN	O	O
respectively	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
.	NN	O	O

Those	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
cancers	NN	O	I-Disease
had	NN	O	O
improved	NN	O	O
survival	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
nonhereditary	NN	O	O
group	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
004	NN	O	O
)	NN	O	O
.	NN	O	O

For	NN	O	O
stage	NN	O	B-Disease
III	NN	O	I-Disease
cancers	NN	O	I-Disease
,	NN	O	O
BRCA	NN	O	O
mutation	NN	O	O
status	NN	O	O
was	NN	O	O
an	NN	O	O
independent	NN	O	O
prognostic	NN	O	O
variable	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
Although	NN	O	O
BRCA	NN	O	B-Disease
-	NN	O	I-Disease
associated	NN	O	I-Disease
hereditary	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
population	NN	O	O
have	NN	O	O
surgical	NN	O	O
and	NN	O	O
pathological	NN	O	O
characteristics	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
sporadic	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
advanced	NN	O	B-Disease
-	NN	O	I-Disease
stage	NN	O	I-Disease
hereditary	NN	O	I-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
survive	NN	O	O
longer	NN	O	O
than	NN	O	O
nonhereditary	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Age	NN	O	O
penetrance	NN	O	O
is	NN	O	O
greater	NN	O	O
for	NN	O	O
BRCA1	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
than	NN	O	I-Disease
for	NN	O	I-Disease
BRCA2	NN	O	I-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
cancers	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
,	NN	O	O
MEFV	NN	O	O
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
early	NN	O	O
leukocyte	NN	O	O
development	NN	O	O
and	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
recessive	NN	O	B-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
episodes	NN	O	O
of	NN	O	O
fever	NN	O	B-Disease
and	NN	O	O
neutrophil	NN	O	B-Disease
-	NN	O	I-Disease
mediated	NN	O	I-Disease
serosal	NN	O	I-Disease
inflammation	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
recently	NN	O	O
identified	NN	O	O
the	NN	O	O
gene	NN	O	O
causing	NN	O	O
FMF	NN	O	B-Disease
,	NN	O	O
designated	NN	O	O
MEFV	NN	O	O
,	NN	O	O
and	NN	O	O
found	NN	O	O
it	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	O
neutrophils	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
functions	NN	O	O
as	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
regulator	NN	O	O
.	NN	O	O

To	NN	O	O
facilitate	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
function	NN	O	O
of	NN	O	O
MEFV	NN	O	O
,	NN	O	O
we	NN	O	O
extended	NN	O	O
our	NN	O	O
previous	NN	O	O
studies	NN	O	O
.	NN	O	O

MEFV	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
-	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
leukocytes	NN	O	O
,	NN	O	O
with	NN	O	O
differential	NN	O	O
expression	NN	O	O
observed	NN	O	O
among	NN	O	O
cells	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

CD34	NN	O	O
hematopoietic	NN	O	O
stem	NN	O	O
-	NN	O	O
cell	NN	O	O
cultures	NN	O	O
induced	NN	O	O
toward	NN	O	O
the	NN	O	O
granulocytic	NN	O	O
lineage	NN	O	O
expressed	NN	O	O
MEFV	NN	O	O
at	NN	O	O
the	NN	O	O
myelocyte	NN	O	O
stage	NN	O	O
,	NN	O	O
concurrently	NN	O	O
with	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

The	NN	O	O
prepromyelocytic	NN	O	O
cell	NN	O	O
line	NN	O	O
HL60	NN	O	O
expressed	NN	O	O
MEFV	NN	O	O
only	NN	O	O
at	NN	O	O
granulocytic	NN	O	O
and	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

MEFV	NN	O	O
was	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
lines	NN	O	O
U937	NN	O	O
and	NN	O	O
THP	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O

Among	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
,	NN	O	O
MEFV	NN	O	O
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
neutrophils	NN	O	O
,	NN	O	O
eosinophils	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
varying	NN	O	O
degrees	NN	O	O
,	NN	O	O
monocytes	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
expression	NN	O	O
,	NN	O	O
complete	NN	O	O
sequencing	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
upstream	NN	O	O
regulatory	NN	O	O
regions	NN	O	O
of	NN	O	O
MEFV	NN	O	O
revealed	NN	O	O
homology	NN	O	O
to	NN	O	O
myeloid	NN	O	O
-	NN	O	O
specific	NN	O	O
promoters	NN	O	O
and	NN	O	O
to	NN	O	O
more	NN	O	O
broadly	NN	O	O
expressed	NN	O	O
inflammatory	NN	O	O
promoter	NN	O	O
elements	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocytes	NN	O	O
with	NN	O	O
the	NN	O	O
proinflammatory	NN	O	O
agents	NN	O	O
interferon	NN	O	O
(	NN	O	O
IFN	NN	O	O
)	NN	O	O
gamma	NN	O	O
,	NN	O	O
tumor	NN	O	B-Disease
necrosis	NN	O	O
factor	NN	O	O
,	NN	O	O
and	NN	O	O
lipopolysaccharide	NN	O	O
induced	NN	O	O
MEFV	NN	O	O
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
antiinflammatory	NN	O	O
cytokines	NN	O	O
interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
4	NN	O	O
,	NN	O	O
IL	NN	O	O
-	NN	O	O
10	NN	O	O
,	NN	O	O
and	NN	O	O
transforming	NN	O	O
growth	NN	O	O
factor	NN	O	O
beta	NN	O	O
inhibited	NN	O	O
such	NN	O	O
expression	NN	O	O
.	NN	O	O

Induction	NN	O	O
by	NN	O	O
IFN	NN	O	O
-	NN	O	O
gamma	NN	O	O
occurred	NN	O	O
rapidly	NN	O	O
and	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

IFN	NN	O	O
-	NN	O	O
alpha	NN	O	O
also	NN	O	O
induced	NN	O	O
MEFV	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
granulocytes	NN	O	O
,	NN	O	O
MEFV	NN	O	O
was	NN	O	O
up	NN	O	O
-	NN	O	O
regulated	NN	O	O
by	NN	O	O
IFN	NN	O	O
-	NN	O	O
gamma	NN	O	O
and	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IFN	NN	O	O
-	NN	O	O
alpha	NN	O	O
and	NN	O	O
colchicine	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
refine	NN	O	O
understanding	NN	O	O
of	NN	O	O
MEFV	NN	O	O
by	NN	O	O
placing	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
myelomonocytic	NN	O	O
-	NN	O	O
specific	NN	O	O
proinflammatory	NN	O	O
pathway	NN	O	O
and	NN	O	O
identifying	NN	O	O
it	NN	O	O
as	NN	O	O
an	NN	O	O
IFN	NN	O	O
-	NN	O	O
gamma	NN	O	O
immediate	NN	O	O
early	NN	O	O
gene	NN	O	O
.	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
and	NN	O	O
structural	NN	O	O
analysis	NN	O	O
of	NN	O	O
missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
N	NN	O	O
-	NN	O	O
acetylgalactosamine	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
sulfate	NN	O	O
sulfatase	NN	O	O
causing	NN	O	O
mucopolysaccharidosis	NN	O	B-Disease
IVA	NN	O	I-Disease
phenotypes	NN	O	O
.	NN	O	O

Mucopolysaccharidosis	NN	O	B-Disease
IVA	NN	O	I-Disease
(	NN	O	O
MPS	NN	O	B-Disease
IVA	NN	O	I-Disease
;	NN	O	O
OMIM	NN	O	O
#	NN	O	O
253000	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
lysosomal	NN	O	B-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
N	NN	O	I-Disease
-	NN	O	I-Disease
acetylgalactosamine	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
sulfate	NN	O	I-Disease
sulfatase	NN	O	I-Disease
(	NN	O	O
GALNS	NN	O	O
)	NN	O	O
,	NN	O	O
has	NN	O	O
variable	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
65	NN	O	O
missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
GALNS	NN	O	O
gene	NN	O	O
from	NN	O	O
MPS	NN	O	B-Disease
IVA	NN	O	I-Disease
patients	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
correlation	NN	O	O
between	NN	O	O
genotype	NN	O	O
and	NN	O	O
phenotype	NN	O	O
has	NN	O	O
remained	NN	O	O
unclear	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
17	NN	O	O
missense	NN	O	O
mutations	NN	O	O
using	NN	O	O
biochemical	NN	O	O
approaches	NN	O	O
and	NN	O	O
32	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
using	NN	O	O
structural	NN	O	O
analyses	NN	O	O
.	NN	O	O

Fifteen	NN	O	O
missense	NN	O	O
mutations	NN	O	O
and	NN	O	O
two	NN	O	O
newly	NN	O	O
engineered	NN	O	O
active	NN	O	O
site	NN	O	O
mutations	NN	O	O
(	NN	O	O
C79S	NN	O	O
,	NN	O	O
C79T	NN	O	O
)	NN	O	O
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
analysis	NN	O	O
.	NN	O	O

Mutant	NN	O	O
proteins	NN	O	O
,	NN	O	O
except	NN	O	O
for	NN	O	O
C79S	NN	O	O
and	NN	O	O
C79T	NN	O	O
,	NN	O	O
were	NN	O	O
destabilized	NN	O	O
and	NN	O	O
detected	NN	O	O
as	NN	O	O
insoluble	NN	O	O
precursor	NN	O	O
forms	NN	O	O
while	NN	O	O
the	NN	O	O
C79S	NN	O	O
and	NN	O	O
C79T	NN	O	O
mutants	NN	O	O
were	NN	O	O
of	NN	O	O
a	NN	O	O
soluble	NN	O	O
mature	NN	O	O
size	NN	O	O
.	NN	O	O

Mutants	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
had	NN	O	O
no	NN	O	O
activity	NN	O	O
.	NN	O	O

Mutants	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
mild	NN	O	O
phenotype	NN	O	O
had	NN	O	O
a	NN	O	O
considerable	NN	O	O
residual	NN	O	O
activity	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
3	NN	O	O
-	NN	O	O
13	NN	O	O
.	NN	O	O
3	NN	O	O
%	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
GALNS	NN	O	O
activity	NN	O	O
)	NN	O	O
.	NN	O	O

Sulfatases	NN	O	O
,	NN	O	O
including	NN	O	O
GALNS	NN	O	O
,	NN	O	O
are	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
gene	NN	O	O
family	NN	O	O
sharing	NN	O	O
an	NN	O	O
extensive	NN	O	O
sequence	NN	O	O
homology	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
a	NN	O	O
tertiary	NN	O	O
structural	NN	O	O
model	NN	O	O
of	NN	O	O
human	NN	O	O
GALNS	NN	O	O
was	NN	O	O
constructed	NN	O	O
from	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
ray	NN	O	O
crystal	NN	O	O
structure	NN	O	O
of	NN	O	O
N	NN	O	O
-	NN	O	O
acetylgalacto	NN	O	O
-	NN	O	O
samine	NN	O	O
-	NN	O	O
4	NN	O	O
-	NN	O	O
sulfatase	NN	O	O
and	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
,	NN	O	O
using	NN	O	O
homology	NN	O	O
modeling	NN	O	O
,	NN	O	O
and	NN	O	O
32	NN	O	O
missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
different	NN	O	O
reasons	NN	O	O
for	NN	O	O
the	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
destruction	NN	O	O
of	NN	O	O
the	NN	O	O
hydrophobic	NN	O	O
core	NN	O	O
or	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
packing	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
removal	NN	O	O
of	NN	O	O
a	NN	O	O
salt	NN	O	O
bridge	NN	O	O
to	NN	O	O
destabilize	NN	O	O
the	NN	O	O
entire	NN	O	O
conformation	NN	O	O
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	O
site	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mild	NN	O	O
mutations	NN	O	O
were	NN	O	O
mostly	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
GALNS	NN	O	O
protein	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
shed	NN	O	O
further	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
of	NN	O	O
MPS	NN	O	B-Disease
IVA	NN	O	I-Disease
and	NN	O	O
structure	NN	O	O
-	NN	O	O
function	NN	O	O
relationship	NN	O	O
in	NN	O	O
the	NN	O	O
sulfatase	NN	O	O
family	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
locus	NN	O	O
and	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
band	NN	O	O
11q22	NN	O	O
cause	NN	O	O
a	NN	O	O
distinctive	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
syndrome	NN	O	I-Disease
in	NN	O	O
homozygotes	NN	O	O
and	NN	O	O
predispose	NN	O	O
heterozygotes	NN	O	O
to	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
ischemic	NN	O	B-Disease
heart	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
early	NN	O	O
mortality	NN	O	O
.	NN	O	O

PCR	NN	O	O
amplification	NN	O	O
from	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
and	NN	O	O
automated	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
(	NN	O	O
66	NN	O	O
exons	NN	O	O
)	NN	O	O
and	NN	O	O
splice	NN	O	O
junctions	NN	O	O
detected	NN	O	O
77	NN	O	O
mutations	NN	O	O
(	NN	O	O
85	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
90	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
chromosomes	NN	O	O
.	NN	O	O

Heteroduplex	NN	O	O
analysis	NN	O	O
detected	NN	O	O
another	NN	O	O
42	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
locus	NN	O	O
.	NN	O	O

Out	NN	O	O
of	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
71	NN	O	O
unique	NN	O	O
mutations	NN	O	O
,	NN	O	O
50	NN	O	O
were	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
a	NN	O	O
single	NN	O	O
family	NN	O	O
,	NN	O	O
and	NN	O	O
51	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
.	NN	O	O

Most	NN	O	O
(	NN	O	O
58	NN	O	O
/	NN	O	O
71	NN	O	O
,	NN	O	O
82	NN	O	O
%	NN	O	O
)	NN	O	O
mutations	NN	O	O
were	NN	O	O
frameshift	NN	O	O
and	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
that	NN	O	O
are	NN	O	O
predicted	NN	O	O
to	NN	O	O
cause	NN	O	O
truncation	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
protein	NN	O	O
;	NN	O	O
the	NN	O	O
less	NN	O	O
common	NN	O	O
mutation	NN	O	O
types	NN	O	O
were	NN	O	O
missense	NN	O	O
(	NN	O	O
9	NN	O	O
/	NN	O	O
71	NN	O	O
,	NN	O	O
13	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
splicing	NN	O	O
(	NN	O	O
3	NN	O	O
/	NN	O	O
71	NN	O	O
,	NN	O	O
4	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
one	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
,	NN	O	O
2546	NN	O	O
3	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
71	NN	O	O
,	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
survival	NN	O	O
and	NN	O	O
height	NN	O	O
distribution	NN	O	O
of	NN	O	O
134	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
correlated	NN	O	O
significantly	NN	O	O
with	NN	O	O
the	NN	O	O
specific	NN	O	O
mutations	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

Patients	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
single	NN	O	O
truncating	NN	O	O
mutation	NN	O	O
,	NN	O	O
typically	NN	O	O
near	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
or	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
2546	NN	O	O
3	NN	O	O
,	NN	O	O
were	NN	O	O
shorter	NN	O	O
and	NN	O	O
had	NN	O	O
significantly	NN	O	O
shorter	NN	O	O
survival	NN	O	O
than	NN	O	O
those	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
splice	NN	O	O
site	NN	O	O
or	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
or	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
two	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
.	NN	O	O

Alterations	NN	O	O
of	NN	O	O
the	NN	O	O
length	NN	O	O
or	NN	O	O
amino	NN	O	O
acid	NN	O	O
composition	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
gene	NN	O	O
product	NN	O	O
affect	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
clinical	NN	O	O
phenotype	NN	O	O
in	NN	O	O
different	NN	O	O
ways	NN	O	O
.	NN	O	O

Mutation	NN	O	O
analysis	NN	O	O
at	NN	O	O
the	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
locus	NN	O	O
may	NN	O	O
help	NN	O	O
estimate	NN	O	O
the	NN	O	O
prognosis	NN	O	O
of	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Isolation	NN	O	O
,	NN	O	O
genomic	NN	O	O
organization	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
rat	NN	O	O
homologs	NN	O	O
of	NN	O	O
MEFV	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
familial	NN	O	B-Disease
mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
.	NN	O	O

Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
recessive	NN	O	B-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
episodes	NN	O	O
of	NN	O	O
fever	NN	O	B-Disease
with	NN	O	O
serositis	NN	O	B-Disease
or	NN	O	O
synovitis	NN	O	B-Disease
.	NN	O	O

Recently	NN	O	O
the	NN	O	O
FMF	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
MEFV	NN	O	O
)	NN	O	O
was	NN	O	O
cloned	NN	O	O
;	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
,	NN	O	O
pyrin	NN	O	O
/	NN	O	O
marenostrin	NN	O	O
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
regulate	NN	O	O
inflammation	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
manuscript	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
rat	NN	O	O
homologs	NN	O	O
of	NN	O	O
MEFV	NN	O	O
.	NN	O	O

The	NN	O	O
murine	NN	O	O
gene	NN	O	O
contains	NN	O	O
ten	NN	O	O
exons	NN	O	O
with	NN	O	O
a	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
2304	NN	O	O
bp	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
rat	NN	O	O
homolog	NN	O	O
has	NN	O	O
nine	NN	O	O
exons	NN	O	O
with	NN	O	O
a	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
2253	NN	O	O
bp	NN	O	O
.	NN	O	O

A	NN	O	O
considerable	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
homology	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
(	NN	O	O
47	NN	O	O
.	NN	O	O
6	NN	O	O
%	NN	O	O
identity	NN	O	O
and	NN	O	O
65	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
similarity	NN	O	O
)	NN	O	O
and	NN	O	O
between	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
rat	NN	O	O
genes	NN	O	O
(	NN	O	O
73	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
identity	NN	O	O
and	NN	O	O
82	NN	O	O
.	NN	O	O
1	NN	O	O
%	NN	O	O
similarity	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
rodent	NN	O	O
proteins	NN	O	O
have	NN	O	O
several	NN	O	O
important	NN	O	O
domains	NN	O	O
and	NN	O	O
signals	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	O
pyrin	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
B	NN	O	O
-	NN	O	O
box	NN	O	O
zinc	NN	O	O
finger	NN	O	O
domain	NN	O	O
,	NN	O	O
Robbins	NN	O	O
-	NN	O	O
Dingwall	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
signal	NN	O	O
,	NN	O	O
and	NN	O	O
coiled	NN	O	O
-	NN	O	O
coil	NN	O	O
domain	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
perhaps	NN	O	O
because	NN	O	O
of	NN	O	O
an	NN	O	O
ancient	NN	O	O
frame	NN	O	O
-	NN	O	O
shift	NN	O	O
mutation	NN	O	O
,	NN	O	O
neither	NN	O	O
the	NN	O	O
mouse	NN	O	O
nor	NN	O	O
the	NN	O	O
rat	NN	O	O
protein	NN	O	O
has	NN	O	O
an	NN	O	O
intact	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
B30	NN	O	O
.	NN	O	O

2	NN	O	O
domain	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
most	NN	O	O
FMF	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	O
MEFV	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
like	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
,	NN	O	O
mouse	NN	O	O
Mefv	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
granulocytes	NN	O	O
but	NN	O	O
not	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
granulocytes	NN	O	O
,	NN	O	O
Mefv	NN	O	O
was	NN	O	O
detected	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
primary	NN	O	O
follicles	NN	O	O
and	NN	O	O
marginal	NN	O	O
zones	NN	O	O
of	NN	O	O
the	NN	O	O
splenic	NN	O	O
white	NN	O	O
pulp	NN	O	O
.	NN	O	O

Mefv	NN	O	O
is	NN	O	O
localized	NN	O	O
on	NN	O	O
mouse	NN	O	O
Chromosome	NN	O	O
(	NN	O	O
Chr	NN	O	O
)	NN	O	O
16	NN	O	O
,	NN	O	O
region	NN	O	O
A3	NN	O	O
-	NN	O	O
B1	NN	O	O
,	NN	O	O
extending	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
synteny	NN	O	O
with	NN	O	O
human	NN	O	O
Chr	NN	O	O
16p13	NN	O	O
.	NN	O	O

3	NN	O	O
.	NN	O	O

Development	NN	O	O
of	NN	O	O
knockout	NN	O	O
and	NN	O	O
knockin	NN	O	O
mouse	NN	O	O
models	NN	O	O
may	NN	O	O
provide	NN	O	O
further	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
functional	NN	O	O
evolution	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

Additional	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
gene	NN	O	O
causing	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
arise	NN	O	O
by	NN	O	O
separate	NN	O	O
integration	NN	O	O
into	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

The	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
gene	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
is	NN	O	O
normally	NN	O	O
present	NN	O	O
at	NN	O	O
chromosome	NN	O	O
Xq22	NN	O	O
.	NN	O	O

Mutations	NN	O	O
and	NN	O	O
duplications	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
PMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
two	NN	O	O
new	NN	O	O
families	NN	O	O
in	NN	O	O
which	NN	O	O
males	NN	O	O
affected	NN	O	O
with	NN	O	O
PMD	NN	O	B-Disease
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
copy	NN	O	O
of	NN	O	O
PLP	NN	O	O
on	NN	O	O
the	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
normal	NN	O	O
copy	NN	O	O
on	NN	O	O
Xq22	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
family	NN	O	O
,	NN	O	O
the	NN	O	O
extra	NN	O	O
copy	NN	O	O
was	NN	O	O
first	NN	O	O
detected	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
of	NN	O	O
the	NN	O	O
AhaII	NN	O	O
dimorphism	NN	O	O
within	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
FISH	NN	O	O
analysis	NN	O	O
showed	NN	O	O
an	NN	O	O
additional	NN	O	O
copy	NN	O	O
of	NN	O	O
PLP	NN	O	O
in	NN	O	O
Xp22	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
although	NN	O	O
no	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
by	NN	O	O
standard	NN	O	O
karyotype	NN	O	O
analysis	NN	O	O
.	NN	O	O

Another	NN	O	O
three	NN	O	O
affected	NN	O	O
males	NN	O	O
from	NN	O	O
the	NN	O	O
family	NN	O	O
had	NN	O	O
similar	NN	O	O
findings	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
second	NN	O	O
unrelated	NN	O	O
family	NN	O	O
with	NN	O	O
signs	NN	O	O
of	NN	O	O
PMD	NN	O	B-Disease
,	NN	O	O
cytogenetic	NN	O	O
analysis	NN	O	O
showed	NN	O	O
a	NN	O	O
pericentric	NN	O	O
inversion	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
inv	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
carried	NN	O	O
by	NN	O	O
several	NN	O	O
affected	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
FISH	NN	O	O
showed	NN	O	O
PLP	NN	O	O
signals	NN	O	O
at	NN	O	O
Xp11	NN	O	O
.	NN	O	O

4	NN	O	O
and	NN	O	O
Xq22	NN	O	O
.	NN	O	O

A	NN	O	O
third	NN	O	O
family	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
reported	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
affected	NN	O	O
members	NN	O	O
had	NN	O	O
an	NN	O	O
extra	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
detected	NN	O	O
at	NN	O	O
Xq26	NN	O	O
in	NN	O	O
a	NN	O	O
chromosome	NN	O	O
with	NN	O	O
an	NN	O	O
otherwise	NN	O	O
normal	NN	O	O
banding	NN	O	O
pattern	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
three	NN	O	O
separate	NN	O	O
families	NN	O	O
in	NN	O	O
which	NN	O	O
PLP	NN	O	O
is	NN	O	O
duplicated	NN	O	O
at	NN	O	O
a	NN	O	O
noncontiguous	NN	O	O
site	NN	O	O
suggests	NN	O	O
that	NN	O	O
such	NN	O	O
duplications	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
relatively	NN	O	O
common	NN	O	O
but	NN	O	O
previously	NN	O	O
undetected	NN	O	O
cause	NN	O	O
of	NN	O	O
genetic	NN	O	B-Disease
disorders	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
exon	NN	O	O
13	NN	O	O
duplication	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
is	NN	O	O
a	NN	O	O
founder	NN	O	O
mutation	NN	O	O
present	NN	O	O
in	NN	O	O
geographically	NN	O	O
diverse	NN	O	O
populations	NN	O	O
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
Exon	NN	O	O
13	NN	O	O
Duplication	NN	O	O
Screening	NN	O	O
Group	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
6	NN	O	O
-	NN	O	O
kb	NN	O	O
duplication	NN	O	O
of	NN	O	O
exon	NN	O	O
13	NN	O	O
,	NN	O	O
which	NN	O	O
creates	NN	O	O
a	NN	O	O
frameshift	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
three	NN	O	O
unrelated	NN	O	O
U	NN	O	O
.	NN	O	O

S	NN	O	O
S	NN	O	O
.	NN	O	O
families	NN	O	O
of	NN	O	O
European	NN	O	O
ancestry	NN	O	O
and	NN	O	O
in	NN	O	O
one	NN	O	O
Portuguese	NN	O	O
family	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
our	NN	O	O
goal	NN	O	O
was	NN	O	O
to	NN	O	O
estimate	NN	O	O
the	NN	O	O
frequency	NN	O	O
and	NN	O	O
geographic	NN	O	O
diversity	NN	O	O
of	NN	O	O
carriers	NN	O	O
of	NN	O	O
this	NN	O	O
duplication	NN	O	O
.	NN	O	O

To	NN	O	O
do	NN	O	O
this	NN	O	O
,	NN	O	O
a	NN	O	O
collaborative	NN	O	O
screening	NN	O	O
study	NN	O	O
was	NN	O	O
set	NN	O	O
up	NN	O	O
that	NN	O	O
involved	NN	O	O
39	NN	O	O
institutions	NN	O	O
from	NN	O	O
19	NN	O	O
countries	NN	O	O
and	NN	O	O
included	NN	O	O
3	NN	O	O
,	NN	O	O
580	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
with	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
and	NN	O	O
934	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
cases	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
11	NN	O	O
additional	NN	O	O
families	NN	O	O
carrying	NN	O	O
this	NN	O	O
mutation	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
Australia	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
Belgium	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
Canada	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
Great	NN	O	O
Britain	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

Haplotyping	NN	O	O
showed	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
derive	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
ancestor	NN	O	O
,	NN	O	O
possibly	NN	O	O
of	NN	O	O
northern	NN	O	O
British	NN	O	O
origin	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
strongly	NN	O	O
advisable	NN	O	O
,	NN	O	O
for	NN	O	O
laboratories	NN	O	O
carrying	NN	O	O
out	NN	O	O
screening	NN	O	O
either	NN	O	O
in	NN	O	O
English	NN	O	O
-	NN	O	O
speaking	NN	O	O
countries	NN	O	O
or	NN	O	O
in	NN	O	O
countries	NN	O	O
with	NN	O	O
historical	NN	O	O
links	NN	O	O
with	NN	O	O
Britain	NN	O	O
,	NN	O	O
to	NN	O	O
include	NN	O	O
within	NN	O	O
their	NN	O	O
BRCA1	NN	O	O
screening	NN	O	O
protocols	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
based	NN	O	O
assay	NN	O	O
described	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
.	NN	O	O

Genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	O
VHL	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	I-Disease
VHL	NN	O	I-Disease
)	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
hereditary	NN	O	B-Disease
tumor	NN	O	I-Disease
syndrome	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
predisposition	NN	O	O
for	NN	O	O
bilateral	NN	O	B-Disease
and	NN	O	I-Disease
multi	NN	O	I-Disease
-	NN	O	I-Disease
centric	NN	O	I-Disease
hemangioblastoma	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
retina	NN	O	O
and	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
pheochromocytoma	NN	O	B-Disease
,	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
cysts	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
kidney	NN	O	I-Disease
,	NN	O	I-Disease
pancreas	NN	O	I-Disease
,	NN	O	I-Disease
and	NN	O	I-Disease
epididymis	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
describe	NN	O	O
five	NN	O	O
families	NN	O	O
for	NN	O	O
which	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
had	NN	O	O
failed	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
family	NN	O	O
-	NN	O	O
specific	NN	O	O
mutation	NN	O	O
.	NN	O	O

Further	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
deletions	NN	O	O
involving	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
four	NN	O	O
families	NN	O	O
,	NN	O	O
partial	NN	O	O
deletions	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
exons	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
fifth	NN	O	O
family	NN	O	O
,	NN	O	O
FISH	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
(	NN	O	O
quantitative	NN	O	O
)	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
is	NN	O	O
a	NN	O	O
sensitive	NN	O	O
method	NN	O	O
for	NN	O	O
detecting	NN	O	O
germline	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
should	NN	O	O
be	NN	O	O
implemented	NN	O	O
in	NN	O	O
routine	NN	O	O
DNA	NN	O	O
diagnosis	NN	O	O
for	NN	O	O
VHL	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
previously	NN	O	O
established	NN	O	O
observation	NN	O	O
that	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
germline	NN	O	O
deletion	NN	O	O
have	NN	O	O
a	NN	O	O
low	NN	O	O
risk	NN	O	O
for	NN	O	O
pheochromocytoma	NN	O	B-Disease
.	NN	O	O

Further	NN	O	O
unraveling	NN	O	O
of	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
in	NN	O	O
VHL	NN	O	B-Disease
disease	NN	O	I-Disease
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
full	NN	O	O
or	NN	O	O
partial	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
exhibit	NN	O	O
a	NN	O	O
phenotype	NN	O	O
with	NN	O	O
a	NN	O	O
preponderance	NN	O	O
of	NN	O	O
central	NN	O	B-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
hemangioblastoma	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Age	NN	O	O
of	NN	O	O
the	NN	O	O
intronic	NN	O	O
GAA	NN	O	O
triplet	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
mutation	NN	O	O
in	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
frequently	NN	O	O
inherited	NN	O	B-Disease
ataxia	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
due	NN	O	O
in	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
cases	NN	O	O
to	NN	O	O
a	NN	O	O
large	NN	O	O
expansion	NN	O	O
of	NN	O	O
an	NN	O	O
intronic	NN	O	O
GAA	NN	O	O
repeat	NN	O	O
.	NN	O	O

Using	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
analysis	NN	O	O
based	NN	O	O
on	NN	O	O
haplotype	NN	O	O
data	NN	O	O
of	NN	O	O
seven	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
frataxin	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
FRDA	NN	O	B-Disease
founding	NN	O	O
mutational	NN	O	O
event	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
is	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
at	NN	O	O
least	NN	O	O
682	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
203	NN	O	O
generations	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
564	NN	O	O
-	NN	O	O
801	NN	O	O
g	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
dating	NN	O	O
which	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
negative	NN	O	O
selection	NN	O	O
and	NN	O	O
provides	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
ancient	NN	O	O
spread	NN	O	O
of	NN	O	O
a	NN	O	O
pre	NN	O	O
-	NN	O	O
mutation	NN	O	O
(	NN	O	O
at	NN	O	O
-	NN	O	O
risk	NN	O	O
alleles	NN	O	O
)	NN	O	O
in	NN	O	O
western	NN	O	O
Europe	NN	O	O
.	NN	O	O
.	NN	O	O

Functional	NN	O	O
link	NN	O	O
between	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
and	NN	O	O
Nijmegen	NN	O	B-Disease
breakage	NN	O	I-Disease
syndrome	NN	O	I-Disease
gene	NN	O	O
products	NN	O	O
.	NN	O	O

Ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
and	NN	O	O
Nijmegen	NN	O	B-Disease
breakage	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
NBS	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
recessive	NN	O	B-Disease
genetic	NN	O	I-Disease
disorders	NN	O	I-Disease
with	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
cancer	NN	O	B-Disease
and	NN	O	O
similar	NN	O	O
cellular	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
,	NN	O	O
designated	NN	O	O
ATM	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
kinases	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminal	NN	O	O
phosphatidylinositol	NN	O	O
3	NN	O	O
-	NN	O	O
kinase	NN	O	O
-	NN	O	O
like	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
NBS1	NN	O	O
protein	NN	O	O
is	NN	O	O
specifically	NN	O	O
mutated	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Nijmegen	NN	O	B-Disease
breakage	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
forms	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	O
repair	NN	O	O
proteins	NN	O	O
Rad50	NN	O	O
and	NN	O	O
Mrel1	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
NBS1	NN	O	O
,	NN	O	O
induced	NN	O	O
by	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
,	NN	O	O
requires	NN	O	O
catalytically	NN	O	O
active	NN	O	O
ATM	NN	O	O
.	NN	O	O

Complexes	NN	O	O
containing	NN	O	O
ATM	NN	O	O
and	NN	O	O
NBS1	NN	O	O
exist	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
both	NN	O	O
untreated	NN	O	O
cells	NN	O	O
and	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
residues	NN	O	O
of	NN	O	O
NBS1	NN	O	O
,	NN	O	O
Ser	NN	O	O
278	NN	O	O
and	NN	O	O
Ser	NN	O	O
343	NN	O	O
that	NN	O	O
are	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
ATM	NN	O	O
and	NN	O	O
whose	NN	O	O
modification	NN	O	O
in	NN	O	O
vivo	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage	NN	O	O
.	NN	O	O

This	NN	O	O
response	NN	O	O
includes	NN	O	O
S	NN	O	O
-	NN	O	O
phase	NN	O	O
checkpoint	NN	O	O
activation	NN	O	O
,	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
NBS1	NN	O	O
/	NN	O	O
Mrel1	NN	O	O
/	NN	O	O
Rad50	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
and	NN	O	O
rescue	NN	O	O
of	NN	O	O
hypersensitivity	NN	O	B-Disease
to	NN	O	I-Disease
ionizing	NN	O	I-Disease
radiation	NN	O	I-Disease
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
biochemical	NN	O	O
link	NN	O	O
between	NN	O	O
cell	NN	O	O
-	NN	O	O
cycle	NN	O	O
checkpoints	NN	O	O
activated	NN	O	O
by	NN	O	O
DNA	NN	O	O
damage	NN	O	O
and	NN	O	O
DNA	NN	O	O
repair	NN	O	O
in	NN	O	O
two	NN	O	O
genetic	NN	O	B-Disease
diseases	NN	O	I-Disease
with	NN	O	O
overlapping	NN	O	O
phenotypes	NN	O	O
.	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
mapping	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
microcephaly	NN	O	B-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
short	NN	O	B-Disease
stature	NN	O	I-Disease
to	NN	O	O
a	NN	O	O
Cohen	NN	O	B-Disease
syndrome	NN	O	I-Disease
region	NN	O	O
on	NN	O	O
8q21	NN	O	O
.	NN	O	O
3	NN	O	O
-	NN	O	O
8q22	NN	O	O
.	NN	O	O
1	NN	O	O
:	NN	O	O
redefining	NN	O	O
a	NN	O	O
clinical	NN	O	O
entity	NN	O	O
.	NN	O	O

A	NN	O	O
syndrome	NN	O	O
of	NN	O	O
microcephaly	NN	O	B-Disease
,	NN	O	O
progressive	NN	O	O
postnatal	NN	O	B-Disease
growth	NN	O	I-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
was	NN	O	O
observed	NN	O	O
in	NN	O	O
two	NN	O	O
brothers	NN	O	O
and	NN	O	O
their	NN	O	O
cousin	NN	O	O
from	NN	O	O
a	NN	O	O
multiply	NN	O	O
consanguineous	NN	O	O
kindred	NN	O	O
of	NN	O	O
Lebanese	NN	O	O
descent	NN	O	O
.	NN	O	O

Hypotonia	NN	O	B-Disease
,	NN	O	O
chorioretinal	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
myopia	NN	O	B-Disease
were	NN	O	O
also	NN	O	O
identified	NN	O	O
.	NN	O	O

The	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
condition	NN	O	O
varied	NN	O	O
among	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
patients	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
distinctive	NN	O	O
facial	NN	O	O
appearance	NN	O	O
,	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
short	NN	O	B-Disease
stature	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
initially	NN	O	O
considered	NN	O	O
clinical	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Cohen	NN	O	B-Disease
syndrome	NN	O	I-Disease
was	NN	O	O
withdrawn	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
genetic	NN	O	O
entity	NN	O	O
was	NN	O	O
assumed	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
mapping	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
assigned	NN	O	O
the	NN	O	O
gene	NN	O	O
to	NN	O	O
a	NN	O	O
26	NN	O	O
.	NN	O	O

8	NN	O	O
-	NN	O	O
cM	NN	O	O
region	NN	O	O
on	NN	O	O
the	NN	O	O
chromosome	NN	O	O
band	NN	O	O
8q21	NN	O	O
.	NN	O	O

3	NN	O	O
-	NN	O	O
22	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
between	NN	O	O
the	NN	O	O
microsatellites	NN	O	O
at	NN	O	O
D8S270	NN	O	O
and	NN	O	O
D8S514	NN	O	O
.	NN	O	O

The	NN	O	O
maximum	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
LOD	NN	O	O
score	NN	O	O
was	NN	O	O
found	NN	O	O
for	NN	O	O
marker	NN	O	O
at	NN	O	O
D8S267	NN	O	O
(	NN	O	O
Zmax	NN	O	O
=	NN	O	O
3	NN	O	O
.	NN	O	O
.	NN	O	O
237	NN	O	O
at	NN	O	O
Omax	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
00	NN	O	O
)	NN	O	O
.	NN	O	O

Intriguingly	NN	O	O
enough	NN	O	O
,	NN	O	O
the	NN	O	O
identified	NN	O	O
gene	NN	O	O
region	NN	O	O
overlaps	NN	O	O
the	NN	O	O
refined	NN	O	O
gene	NN	O	O
region	NN	O	O
for	NN	O	O
Cohen	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
COH1	NN	O	O
)	NN	O	O
[	NN	O	O
Kolehmainen	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1997	NN	O	O
Euro	NN	O	O
J	NN	O	O
Hum	NN	O	O
Genet	NN	O	O
5	NN	O	O
206	NN	O	O
-	NN	O	O
213	NN	O	O
]	NN	O	O
.	NN	O	O

This	NN	O	O
fact	NN	O	O
encourages	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
described	NN	O	O
kindred	NN	O	O
segregates	NN	O	O
for	NN	O	O
a	NN	O	O
variant	NN	O	O
of	NN	O	O
Cohen	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
suggests	NN	O	O
a	NN	O	O
redefinition	NN	O	O
of	NN	O	O
its	NN	O	O
phenotype	NN	O	O

Polymorphisms	NN	O	O
of	NN	O	O
the	NN	O	O
CYP2D6	NN	O	O
gene	NN	O	O
increase	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
.	NN	O	O

Ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
and	NN	O	O
highly	NN	O	O
familial	NN	O	O
rheumatic	NN	O	B-Disease
disorder	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
sibling	NN	O	O
recurrence	NN	O	O
risk	NN	O	O
ratio	NN	O	O
for	NN	O	O
the	NN	O	O
disease	NN	O	O
is	NN	O	O
63	NN	O	O
and	NN	O	O
heritability	NN	O	O
assessed	NN	O	O
in	NN	O	O
twins	NN	O	O
>	NN	O	O
90	NN	O	O
%	NN	O	O
.	NN	O	O

Although	NN	O	O
MHC	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
HLA	NN	O	O
-	NN	O	O
B27	NN	O	O
,	NN	O	O
contribute	NN	O	O
only	NN	O	O
20	NN	O	O
-	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
risk	NN	O	O
for	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
no	NN	O	O
non	NN	O	O
-	NN	O	O
MHC	NN	O	O
gene	NN	O	O
has	NN	O	O
yet	NN	O	O
been	NN	O	O
convincingly	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
influence	NN	O	O
either	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
the	NN	O	O
disease	NN	O	O
or	NN	O	O
its	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
.	NN	O	O

Previous	NN	O	O
linkage	NN	O	O
and	NN	O	O
association	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
susceptibility	NN	O	O
gene	NN	O	O
for	NN	O	O
AS	NN	O	B-Disease
close	NN	O	O
to	NN	O	O
,	NN	O	O
or	NN	O	O
within	NN	O	O
,	NN	O	O
the	NN	O	O
cytochrome	NN	O	O
P450	NN	O	O
2D6	NN	O	O
gene	NN	O	O
(	NN	O	O
CYP2D6	NN	O	O
,	NN	O	O
debrisoquine	NN	O	O
hydroxylase	NN	O	O
)	NN	O	O
located	NN	O	O
at	NN	O	O
chromosome	NN	O	O
22q13	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
a	NN	O	O
linkage	NN	O	O
study	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22	NN	O	O
in	NN	O	O
200	NN	O	O
families	NN	O	O
with	NN	O	O
AS	NN	O	B-Disease
affected	NN	O	O
sibling	NN	O	O
-	NN	O	O
pairs	NN	O	O
.	NN	O	O

Association	NN	O	O
of	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
CYP2D6	NN	O	O
gene	NN	O	O
was	NN	O	O
examined	NN	O	O
by	NN	O	O
both	NN	O	O
case	NN	O	O
-	NN	O	O
control	NN	O	O
and	NN	O	O
within	NN	O	O
-	NN	O	O
family	NN	O	O
means	NN	O	O
.	NN	O	O

For	NN	O	O
case	NN	O	O
-	NN	O	O
control	NN	O	O
studies	NN	O	O
,	NN	O	O
617	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
with	NN	O	O
AS	NN	O	B-Disease
(	NN	O	O
361	NN	O	O
probands	NN	O	O
from	NN	O	O
sibling	NN	O	O
-	NN	O	O
pair	NN	O	O
and	NN	O	O
parent	NN	O	O
-	NN	O	O
case	NN	O	O
trio	NN	O	O
families	NN	O	O
and	NN	O	O
256	NN	O	O
unrelated	NN	O	O
non	NN	O	O
-	NN	O	O
familial	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
)	NN	O	O
and	NN	O	O
402	NN	O	O
healthy	NN	O	O
ethnically	NN	O	O
matched	NN	O	O
controls	NN	O	O
were	NN	O	O
employed	NN	O	O
.	NN	O	O

For	NN	O	O
within	NN	O	O
-	NN	O	O
family	NN	O	O
association	NN	O	O
studies	NN	O	O
,	NN	O	O
361	NN	O	O
families	NN	O	O
including	NN	O	O
161	NN	O	O
parent	NN	O	O
-	NN	O	O
case	NN	O	O
trios	NN	O	O
and	NN	O	O
200	NN	O	O
affected	NN	O	O
sibling	NN	O	O
-	NN	O	O
pair	NN	O	O
families	NN	O	O
were	NN	O	O
employed	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
for	NN	O	O
poor	NN	O	O
metabolizer	NN	O	O
alleles	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
AS	NN	O	B-Disease
.	NN	O	O

Heterozygosity	NN	O	O
for	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
poor	NN	O	O
metabolizer	NN	O	O
allele	NN	O	O
(	NN	O	O
CYP2D6	NN	O	O
*	NN	O	O
4	NN	O	O
)	NN	O	O
was	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
AS	NN	O	B-Disease
.	NN	O	O

Significant	NN	O	O
within	NN	O	O
-	NN	O	O
family	NN	O	O
association	NN	O	O
of	NN	O	O
CYP2D6	NN	O	O
*	NN	O	O
4	NN	O	O
alleles	NN	O	O
and	NN	O	O
AS	NN	O	B-Disease
was	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

Weak	NN	O	O
linkage	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
between	NN	O	O
CYP2D6	NN	O	O
and	NN	O	O
AS	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
postulate	NN	O	O
that	NN	O	O
altered	NN	O	O
metabolism	NN	O	O
of	NN	O	O
a	NN	O	O
natural	NN	O	O
toxin	NN	O	O
or	NN	O	O
antigen	NN	O	O
by	NN	O	O
the	NN	O	O
CYP2D6	NN	O	O
gene	NN	O	O
may	NN	O	O
increase	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
AS	NN	O	B-Disease
.	NN	O	O

Functional	NN	O	O
differences	NN	O	O
of	NN	O	O
the	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
phenotypic	NN	O	O
variation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
(	NN	O	O
DFNB4	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
PDS	NN	O	B-Disease
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
transmembrane	NN	O	O
protein	NN	O	O
,	NN	O	O
known	NN	O	O
as	NN	O	O
pendrin	NN	O	O
,	NN	O	O
which	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transporter	NN	O	O
of	NN	O	O
iodide	NN	O	O
and	NN	O	O
chloride	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
non	NN	O	I-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
DFNB4	NN	O	B-Disease
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
7q31	NN	O	O
.	NN	O	O

A	NN	O	O
screen	NN	O	O
of	NN	O	O
20	NN	O	O
individuals	NN	O	O
from	NN	O	O
the	NN	O	O
midwestern	NN	O	O
USA	NN	O	O
with	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
and	NN	O	O
dilated	NN	O	O
vestibular	NN	O	O
aqueducts	NN	O	O
identified	NN	O	O
three	NN	O	O
people	NN	O	O
(	NN	O	O
15	NN	O	O
%	NN	O	O
)	NN	O	O
with	NN	O	O
PDS	NN	O	B-Disease
mutations	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
PDS	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
differ	NN	O	O
functionally	NN	O	O
from	NN	O	O
PDS	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
compared	NN	O	O
three	NN	O	O
common	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
allele	NN	O	O
variants	NN	O	O
(	NN	O	O
L236P	NN	O	O
,	NN	O	O
T416P	NN	O	O
and	NN	O	O
E384G	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
three	NN	O	O
PDS	NN	O	B-Disease
mutations	NN	O	O
reported	NN	O	O
only	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
syndromic	NN	O	I-Disease
hearing	NN	O	I-Disease
loss	NN	O	I-Disease
(	NN	O	O
V480D	NN	O	O
,	NN	O	O
V653A	NN	O	O
and	NN	O	O
I490L	NN	O	O
/	NN	O	O
G497S	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
associated	NN	O	O
with	NN	O	O
Pendred	NN	O	B-Disease
syndrome	NN	O	I-Disease
have	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
pendrin	NN	O	O
-	NN	O	O
induced	NN	O	O
chloride	NN	O	O
and	NN	O	O
iodide	NN	O	O
transport	NN	O	O
,	NN	O	O
while	NN	O	O
alleles	NN	O	O
unique	NN	O	O
to	NN	O	O
people	NN	O	O
with	NN	O	O
DFNB4	NN	O	B-Disease
are	NN	O	O
able	NN	O	O
to	NN	O	O
transport	NN	O	O
both	NN	O	O
iodide	NN	O	O
and	NN	O	O
chloride	NN	O	O
,	NN	O	O
albeit	NN	O	O
at	NN	O	O
a	NN	O	O
much	NN	O	O
lower	NN	O	O
level	NN	O	O
than	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
pendrin	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
this	NN	O	O
residual	NN	O	O
level	NN	O	O
of	NN	O	O
anion	NN	O	O
transport	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
eliminate	NN	O	O
or	NN	O	O
postpone	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
goiter	NN	O	B-Disease
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
DFNB4	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
pendrin	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
thyroid	NN	O	O
in	NN	O	O
which	NN	O	O
pendrin	NN	O	O
transports	NN	O	O
iodide	NN	O	O
across	NN	O	O
the	NN	O	O
apical	NN	O	O
membrane	NN	O	O
of	NN	O	O
the	NN	O	O
thyrocyte	NN	O	O
into	NN	O	O
the	NN	O	O
colloid	NN	O	O
space	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
factor	NN	O	O
IX	NN	O	O
gene	NN	O	O
as	NN	O	O
germline	NN	O	O
mutagen	NN	O	O
test	NN	O	O
:	NN	O	O
samples	NN	O	O
from	NN	O	O
Mainland	NN	O	O
China	NN	O	O
have	NN	O	O
the	NN	O	O
putatively	NN	O	O
endogenous	NN	O	O
pattern	NN	O	O
of	NN	O	O
mutation	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
are	NN	O	O
the	NN	O	O
major	NN	O	O
source	NN	O	O
of	NN	O	O
genetic	NN	O	O
variation	NN	O	O
that	NN	O	O
allows	NN	O	O
a	NN	O	O
species	NN	O	O
to	NN	O	O
evolve	NN	O	O
over	NN	O	O
time	NN	O	O
but	NN	O	O
at	NN	O	O
the	NN	O	O
cost	NN	O	O
of	NN	O	O
Mendelian	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
genetic	NN	O	O
predisposition	NN	O	O
to	NN	O	O
multifactorial	NN	O	B-Disease
diseases	NN	O	I-Disease
.	NN	O	O

Previous	NN	O	O
analyses	NN	O	O
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
factor	NN	O	O
IX	NN	O	O
gene	NN	O	O
(	NN	O	O
F9	NN	O	O
)	NN	O	O
is	NN	O	O
similar	NN	O	O
among	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
ethnically	NN	O	O
and	NN	O	O
geographically	NN	O	O
diverse	NN	O	O
populations	NN	O	O
and	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
ancient	NN	O	O
pattern	NN	O	O
that	NN	O	O
has	NN	O	O
shaped	NN	O	O
the	NN	O	O
mammalian	NN	O	O
genome	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
compare	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
hemophilia	NN	O	B-Disease
B	NN	O	I-Disease
patients	NN	O	O
from	NN	O	O
Mainland	NN	O	O
China	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
66	NN	O	O
)	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
U	NN	O	O
.	NN	O	O

S	NN	O	O
.	NN	O	O

Caucasians	NN	O	O
,	NN	O	O
Blacks	NN	O	O
,	NN	O	O
and	NN	O	O
Mexican	NN	O	O
Hispanics	NN	O	O
and	NN	O	O
stratify	NN	O	O
by	NN	O	O
disease	NN	O	O
severity	NN	O	O
and	NN	O	O
ethnicity	NN	O	O
.	NN	O	O

The	NN	O	O
similar	NN	O	O
pattern	NN	O	O
of	NN	O	O
germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
all	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
studied	NN	O	O
to	NN	O	O
date	NN	O	O
provides	NN	O	O
additional	NN	O	O
data	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
inference	NN	O	O
that	NN	O	O
endogenous	NN	O	O
processes	NN	O	O
predominate	NN	O	O
in	NN	O	O
germline	NN	O	O
mutations	NN	O	O
.	NN	O	O

Fas	NN	O	O
preassociation	NN	O	O
required	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
signaling	NN	O	O
and	NN	O	O
dominant	NN	O	O
inhibition	NN	O	O
by	NN	O	O
pathogenic	NN	O	O
mutations	NN	O	O
.	NN	O	O

Heterozygous	NN	O	O
mutations	NN	O	O
encoding	NN	O	O
abnormal	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
death	NN	O	O
receptor	NN	O	O
Fas	NN	O	O
dominantly	NN	O	O
interfere	NN	O	O
with	NN	O	O
Fas	NN	O	O
-	NN	O	O
induced	NN	O	O
lymphocyte	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	O
autoimmune	NN	O	B-Disease
lymphoproliferative	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
effect	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
depending	NN	O	O
on	NN	O	O
ligand	NN	O	O
-	NN	O	O
induced	NN	O	O
receptor	NN	O	O
oligomerization	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
stem	NN	O	O
from	NN	O	O
ligand	NN	O	O
-	NN	O	O
independent	NN	O	O
interaction	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
and	NN	O	O
mutant	NN	O	O
Fas	NN	O	O
receptors	NN	O	O
through	NN	O	O
a	NN	O	O
specific	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
extracellular	NN	O	O
domain	NN	O	O
.	NN	O	O

Preassociated	NN	O	O
Fas	NN	O	O
complexes	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
living	NN	O	O
cells	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
fluorescence	NN	O	O
resonance	NN	O	O
energy	NN	O	O
transfer	NN	O	O
between	NN	O	O
variants	NN	O	O
of	NN	O	O
green	NN	O	O
fluorescent	NN	O	O
protein	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
formation	NN	O	O
of	NN	O	O
preassociated	NN	O	O
receptor	NN	O	O
complexes	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
Fas	NN	O	O
signaling	NN	O	O
and	NN	O	O
dominant	NN	O	O
interference	NN	O	O
in	NN	O	O
human	NN	O	O
disease	NN	O	O
.	NN	O	O
.	NN	O	O

Determination	NN	O	O
of	NN	O	O
carrier	NN	O	O
status	NN	O	O
for	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
by	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
of	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
mononuclear	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
protein	NN	O	O
(	NN	O	O
WASP	NN	O	O
)	NN	O	O
gene	NN	O	O
on	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Previous	NN	O	O
study	NN	O	O
disclosed	NN	O	O
that	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
of	NN	O	O
intracellular	NN	O	O
WASP	NN	O	O
expression	NN	O	O
(	NN	O	O
FCM	NN	O	O
-	NN	O	O
WASP	NN	O	O
analysis	NN	O	O
)	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
was	NN	O	O
useful	NN	O	O
for	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	O
from	NN	O	O
all	NN	O	O
WAS	NN	O	B-Disease
patients	NN	O	O
showed	NN	O	O
WASPdim	NN	O	O
instead	NN	O	O
of	NN	O	O
WASPbright	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
FCM	NN	O	O
-	NN	O	O
WASP	NN	O	O
analysis	NN	O	O
in	NN	O	O
monocytes	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
carrier	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

Monocytes	NN	O	O
from	NN	O	O
all	NN	O	O
nine	NN	O	O
WAS	NN	O	B-Disease
carriers	NN	O	O
showed	NN	O	O
varied	NN	O	O
population	NN	O	O
of	NN	O	O
WASPdim	NN	O	O
together	NN	O	O
with	NN	O	O
WASPbright	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
control	NN	O	O
individuals	NN	O	O
possessed	NN	O	O
the	NN	O	O
WASPdim	NN	O	O
population	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
all	NN	O	O
the	NN	O	O
carriers	NN	O	O
except	NN	O	O
two	NN	O	O
lacked	NN	O	O
the	NN	O	O
WASPdim	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
of	NN	O	O
the	NN	O	O
WASPdim	NN	O	O
population	NN	O	O
in	NN	O	O
monocytes	NN	O	O
and	NN	O	O
lymphocytes	NN	O	O
observed	NN	O	O
in	NN	O	O
WAS	NN	O	B-Disease
carriers	NN	O	O
suggests	NN	O	O
that	NN	O	O
WASP	NN	O	O
plays	NN	O	O
a	NN	O	O
more	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
than	NN	O	O
in	NN	O	O
that	NN	O	O
of	NN	O	O
monocytes	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
skewed	NN	O	O
X	NN	O	O
-	NN	O	O
chromosomal	NN	O	O
inactivation	NN	O	O
pattern	NN	O	O
observed	NN	O	O
in	NN	O	O
WAS	NN	O	B-Disease
carrier	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
cells	NN	O	O
is	NN	O	O
not	NN	O	O
fixed	NN	O	O
at	NN	O	O
the	NN	O	O
hemopoietic	NN	O	O
stem	NN	O	O
cell	NN	O	O
level	NN	O	O
but	NN	O	O
progresses	NN	O	O
after	NN	O	O
the	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O
.	NN	O	O

Restoration	NN	O	O
of	NN	O	O
photoreceptor	NN	O	O
ultrastructure	NN	O	O
and	NN	O	O
function	NN	O	O
in	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
slow	NN	O	O
mice	NN	O	O
by	NN	O	O
gene	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
Prph2	NN	O	O
encodes	NN	O	O
a	NN	O	O
photoreceptor	NN	O	O
-	NN	O	O
specific	NN	O	O
membrane	NN	O	O
glycoprotein	NN	O	O
,	NN	O	O
peripherin	NN	O	O
-	NN	O	O
2	NN	O	O
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
peripherin	NN	O	O
/	NN	O	O
rds	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
rims	NN	O	O
of	NN	O	O
photoreceptor	NN	O	O
outer	NN	O	O
segment	NN	O	O
discs	NN	O	O
in	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
rom	NN	O	O
-	NN	O	O
1	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
complex	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
discs	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
renewed	NN	O	O
constantly	NN	O	O
throughout	NN	O	O
life	NN	O	O
,	NN	O	O
and	NN	O	O
which	NN	O	O
contain	NN	O	O
the	NN	O	O
visual	NN	O	O
pigments	NN	O	O
necessary	NN	O	O
for	NN	O	O
photon	NN	O	O
capture	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
Prph2	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
photoreceptor	NN	O	B-Disease
dystrophies	NN	O	I-Disease
,	NN	O	O
including	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
retinitis	NN	O	I-Disease
pigmentosa	NN	O	I-Disease
and	NN	O	O
macular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
common	NN	O	O
feature	NN	O	O
of	NN	O	O
these	NN	O	O
diseases	NN	O	O
is	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
photoreceptor	NN	O	O
function	NN	O	O
,	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
slow	NN	O	O
(	NN	O	O
rds	NN	O	O
or	NN	O	O
Prph2	NN	O	O
Rd2	NN	O	O
/	NN	O	O
Rd2	NN	O	O
)	NN	O	O
mouse	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
null	NN	O	O
mutation	NN	O	O
in	NN	O	O
Prph2	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
complete	NN	O	O
failure	NN	O	O
to	NN	O	O
develop	NN	O	O
photoreceptor	NN	O	O
discs	NN	O	O
and	NN	O	O
outer	NN	O	O
segments	NN	O	O
,	NN	O	O
downregulation	NN	O	O
of	NN	O	O
rhodopsin	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
loss	NN	O	O
of	NN	O	O
photoreceptor	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
electroretinograms	NN	O	O
(	NN	O	O
ERGs	NN	O	O
)	NN	O	O
of	NN	O	O
Prph2Rd2	NN	O	O
/	NN	O	O
Rd2	NN	O	O
mice	NN	O	O
have	NN	O	O
greatly	NN	O	O
diminished	NN	O	O
a	NN	O	O
-	NN	O	O
wave	NN	O	O
and	NN	O	O
b	NN	O	O
-	NN	O	O
wave	NN	O	O
amplitudes	NN	O	O
,	NN	O	O
which	NN	O	O
decline	NN	O	O
to	NN	O	O
virtually	NN	O	O
undetectable	NN	O	O
concentrations	NN	O	O
by	NN	O	O
two	NN	O	O
months	NN	O	O
.	NN	O	O

Subretinal	NN	O	O
injection	NN	O	O
of	NN	O	O
recombinant	NN	O	O
adeno	NN	O	O
-	NN	O	O
associated	NN	O	O
virus	NN	O	O
(	NN	O	O
AAV	NN	O	O
)	NN	O	O
encoding	NN	O	O
a	NN	O	O
Prph2	NN	O	O
transgene	NN	O	O
results	NN	O	O
in	NN	O	O
stable	NN	O	O
generation	NN	O	O
of	NN	O	O
outer	NN	O	O
segment	NN	O	O
structures	NN	O	O
and	NN	O	O
formation	NN	O	O
of	NN	O	O
new	NN	O	O
stacks	NN	O	O
of	NN	O	O
discs	NN	O	O
containing	NN	O	O
both	NN	O	O
perpherin	NN	O	O
-	NN	O	O
2	NN	O	O
and	NN	O	O
rhodopsin	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
many	NN	O	O
cases	NN	O	O
are	NN	O	O
morphologically	NN	O	O
similar	NN	O	O
to	NN	O	O
normal	NN	O	O
outer	NN	O	O
segments	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
re	NN	O	O
-	NN	O	O
establishment	NN	O	O
of	NN	O	O
the	NN	O	O
structural	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
photoreceptor	NN	O	O
layer	NN	O	O
also	NN	O	O
results	NN	O	O
in	NN	O	O
electrophysiological	NN	O	O
correction	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
a	NN	O	O
complex	NN	O	O
ultrastructural	NN	O	O
cell	NN	O	O
defect	NN	O	O
can	NN	O	O
be	NN	O	O
corrected	NN	O	O
both	NN	O	O
morphologically	NN	O	O
and	NN	O	O
functionally	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
gene	NN	O	O
transfer	NN	O	O
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
fibrinogen	NN	O	O
aalpha	NN	O	O
gene	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
cases	NN	O	O
of	NN	O	O
congenital	NN	O	B-Disease
afibrinogenemia	NN	O	I-Disease
.	NN	O	O

Congenital	NN	O	B-Disease
afibrinogenemia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
,	NN	O	O
autosomal	NN	O	B-Disease
,	NN	O	I-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
complete	NN	O	O
absence	NN	O	O
of	NN	O	O
detectable	NN	O	O
fibrinogen	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
identified	NN	O	O
the	NN	O	O
first	NN	O	O
causative	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
nonconsanguineous	NN	O	O
Swiss	NN	O	O
family	NN	O	O
;	NN	O	O
the	NN	O	O
4	NN	O	O
affected	NN	O	O
persons	NN	O	O
have	NN	O	O
homozygous	NN	O	O
deletions	NN	O	O
of	NN	O	O
approximately	NN	O	O
11	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
fibrinogen	NN	O	O
alpha	NN	O	O
(	NN	O	O
FGA	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
data	NN	O	O
implied	NN	O	O
that	NN	O	O
these	NN	O	O
deletions	NN	O	O
occurred	NN	O	O
on	NN	O	O
distinct	NN	O	O
ancestral	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
region	NN	O	O
may	NN	O	O
be	NN	O	O
susceptible	NN	O	O
to	NN	O	O
deletion	NN	O	O
by	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
.	NN	O	O

We	NN	O	O
subsequently	NN	O	O
showed	NN	O	O
that	NN	O	O
all	NN	O	O
the	NN	O	O
deletions	NN	O	O
were	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
base	NN	O	O
pair	NN	O	O
and	NN	O	O
probably	NN	O	O
resulted	NN	O	O
from	NN	O	O
a	NN	O	O
nonhomologous	NN	O	O
recombination	NN	O	O
mediated	NN	O	O
by	NN	O	O
7	NN	O	O
-	NN	O	O
bp	NN	O	O
direct	NN	O	O
repeats	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
collected	NN	O	O
data	NN	O	O
on	NN	O	O
13	NN	O	O
additional	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
causative	NN	O	O
mutations	NN	O	O
and	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
the	NN	O	O
11	NN	O	O
-	NN	O	O
kb	NN	O	O
deletion	NN	O	O
.	NN	O	O

A	NN	O	O
common	NN	O	O
recurrent	NN	O	O
mutation	NN	O	O
,	NN	O	O
at	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
FGA	NN	O	O
intron	NN	O	O
4	NN	O	O
(	NN	O	O
IVS4	NN	O	O
+	NN	O	O
1	NN	O	O
G	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
,	NN	O	O
accounted	NN	O	O
for	NN	O	O
14	NN	O	O
of	NN	O	O
the	NN	O	O
26	NN	O	O
(	NN	O	O
54	NN	O	O
%	NN	O	O
)	NN	O	O
alleles	NN	O	O
.	NN	O	O

One	NN	O	O
patient	NN	O	O
was	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
previously	NN	O	O
identified	NN	O	O
deletion	NN	O	O
.	NN	O	O

Three	NN	O	O
more	NN	O	O
frameshift	NN	O	O
mutations	NN	O	O
,	NN	O	O
2	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
second	NN	O	O
splice	NN	O	O
site	NN	O	O
mutation	NN	O	O
were	NN	O	O
also	NN	O	O
identified	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
86	NN	O	O
%	NN	O	O
of	NN	O	O
afibrinogenemia	NN	O	B-Disease
alleles	NN	O	O
analyzed	NN	O	O
to	NN	O	O
date	NN	O	O
have	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
of	NN	O	O
FGA	NN	O	O
,	NN	O	O
though	NN	O	O
mutations	NN	O	O
in	NN	O	O
all	NN	O	O
3	NN	O	O
fibrinogen	NN	O	O
genes	NN	O	O
,	NN	O	O
FGG	NN	O	O
,	NN	O	O
FGA	NN	O	O
,	NN	O	O
and	NN	O	O
FGB	NN	O	O
,	NN	O	O
might	NN	O	O
be	NN	O	O
predicted	NN	O	O
to	NN	O	O
cause	NN	O	O
congenital	NN	O	B-Disease
afibrinogenemia	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
:	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
CUG	NN	O	O
triplet	NN	O	O
repeats	NN	O	O
in	NN	O	O
splicing	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
DMPK	NN	O	O
exon	NN	O	O
and	NN	O	O
altered	NN	O	O
cytoplasmic	NN	O	O
DMPK	NN	O	O
mRNA	NN	O	O
isoform	NN	O	O
ratios	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
expansion	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
UTR	NN	O	O
of	NN	O	O
the	NN	O	O
DMPK	NN	O	O
gene	NN	O	O
causes	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
four	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
factors	NN	O	O
-	NN	O	O
-	NN	O	O
hnRNP	NN	O	O
C	NN	O	O
,	NN	O	O
U2AF	NN	O	O
(	NN	O	O
U2	NN	O	O
auxiliary	NN	O	O
factor	NN	O	O
)	NN	O	O
,	NN	O	O
PTB	NN	O	O
(	NN	O	O
polypyrimidine	NN	O	O
tract	NN	O	O
binding	NN	O	O
protein	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
PSF	NN	O	O
(	NN	O	O
PTB	NN	O	O
associated	NN	O	O
splicing	NN	O	O
factor	NN	O	O
)	NN	O	O
-	NN	O	O
-	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
two	NN	O	O
short	NN	O	O
regions	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
(	NN	O	O
CUG	NN	O	O
)	NN	O	O
n	NN	O	O
,	NN	O	O
and	NN	O	O
found	NN	O	O
a	NN	O	O
novel	NN	O	O
3	NN	O	O
DMPK	NN	O	O
exon	NN	O	O
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
mRNA	NN	O	O
lacking	NN	O	O
the	NN	O	O
repeats	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
(	NN	O	O
CUG	NN	O	O
)	NN	O	O
n	NN	O	O
is	NN	O	O
an	NN	O	O
essential	NN	O	O
cis	NN	O	O
acting	NN	O	O
element	NN	O	O
for	NN	O	O
this	NN	O	O
splicing	NN	O	O
event	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
(	NN	O	O
CUG	NN	O	O
)	NN	O	O
n	NN	O	O
containing	NN	O	O
mRNAs	NN	O	O
,	NN	O	O
the	NN	O	O
novel	NN	O	O
isoform	NN	O	O
is	NN	O	O
not	NN	O	O
retained	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
cells	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
imbalances	NN	O	O
in	NN	O	O
relative	NN	O	O
levels	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	O
DMPK	NN	O	O
mRNA	NN	O	O
isoforms	NN	O	O
and	NN	O	O
a	NN	O	O
new	NN	O	O
dominant	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
on	NN	O	O
DMPK	NN	O	O
.	NN	O	O
.	NN	O	O

Expression	NN	O	O
and	NN	O	O
imprinting	NN	O	O
of	NN	O	O
MAGEL2	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
homologous	NN	O	O
murine	NN	O	O
imprinting	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
imprinted	NN	O	O
genes	NN	O	O
in	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
.	NN	O	O

Affected	NN	O	O
individuals	NN	O	O
exhibit	NN	O	O
neonatal	NN	O	B-Disease
hypotonia	NN	O	I-Disease
,	NN	O	O
developmental	NN	O	B-Disease
delay	NN	O	I-Disease
and	NN	O	O
childhood	NN	O	B-Disease
-	NN	O	I-Disease
onset	NN	O	I-Disease
obesity	NN	O	I-Disease
.	NN	O	O

Necdin	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
neurons	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
only	NN	O	O
PWS	NN	O	B-Disease
candidate	NN	O	O
gene	NN	O	O
to	NN	O	O
reduce	NN	O	O
viability	NN	O	O
when	NN	O	O
disrupted	NN	O	O
in	NN	O	O
a	NN	O	O
mouse	NN	O	O
model	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
MAGEL2	NN	O	O
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
NDNL1	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
with	NN	O	O
51	NN	O	O
%	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
necdin	NN	O	O
and	NN	O	O
located	NN	O	O
41	NN	O	O
kb	NN	O	O
distal	NN	O	O
to	NN	O	O
NDN	NN	O	O
in	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
deletion	NN	O	O
region	NN	O	O
.	NN	O	O

MAGEL2	NN	O	O
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
brain	NN	O	O
,	NN	O	O
the	NN	O	O
primary	NN	O	O
tissue	NN	O	O
affected	NN	O	O
in	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
in	NN	O	O
several	NN	O	O
fetal	NN	O	O
tissues	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

MAGEL2	NN	O	O
is	NN	O	O
imprinted	NN	O	O
with	NN	O	O
monoallelic	NN	O	O
expression	NN	O	O
in	NN	O	O
control	NN	O	O
brain	NN	O	O
,	NN	O	O
and	NN	O	O
paternal	NN	O	O
-	NN	O	O
only	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
its	NN	O	O
lack	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
brain	NN	O	O
from	NN	O	O
a	NN	O	O
PWS	NN	O	B-Disease
-	NN	O	O
affected	NN	O	O
individual	NN	O	O
.	NN	O	O

The	NN	O	O
orthologous	NN	O	O
mouse	NN	O	O
gene	NN	O	O
(	NN	O	O
Magel2	NN	O	O
)	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
150	NN	O	O
kb	NN	O	O
of	NN	O	O
NDN	NN	O	O
,	NN	O	O
is	NN	O	O
imprinted	NN	O	O
with	NN	O	O
paternal	NN	O	O
-	NN	O	O
only	NN	O	O
expression	NN	O	O
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
late	NN	O	O
developmental	NN	O	O
stages	NN	O	O
and	NN	O	O
adult	NN	O	O
brain	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
northern	NN	O	O
blotting	NN	O	O
,	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
and	NN	O	O
whole	NN	O	O
-	NN	O	O
mount	NN	O	O
RNA	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

Magel2	NN	O	O
distribution	NN	O	O
partially	NN	O	O
overlaps	NN	O	O
that	NN	O	O
of	NN	O	O
NDN	NN	O	O
,	NN	O	O
with	NN	O	O
strong	NN	O	O
expression	NN	O	O
being	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
in	NN	O	O
mid	NN	O	O
-	NN	O	O
gestation	NN	O	O
mouse	NN	O	O
embryos	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
,	NN	O	O
although	NN	O	O
loss	NN	O	O
of	NN	O	O
necdin	NN	O	O
expression	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
neonatal	NN	O	O
presentation	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
,	NN	O	O
loss	NN	O	O
of	NN	O	O
MAGEL2	NN	O	O
may	NN	O	O
be	NN	O	O
critical	NN	O	O
to	NN	O	O
abnormalities	NN	O	B-Disease
in	NN	O	I-Disease
brain	NN	O	I-Disease
development	NN	O	I-Disease
and	NN	O	O
dysmorphic	NN	O	B-Disease
features	NN	O	I-Disease
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Retinoschisin	NN	O	O
,	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
retinoschisis	NN	O	I-Disease
protein	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
secreted	NN	O	O
photoreceptor	NN	O	O
protein	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
and	NN	O	O
released	NN	O	O
by	NN	O	O
Weri	NN	O	O
-	NN	O	O
Rb1	NN	O	O
cells	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
retinoschisis	NN	O	I-Disease
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
microcystic	NN	O	O
-	NN	O	O
like	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
macular	NN	O	O
region	NN	O	O
and	NN	O	O
schisis	NN	O	O
within	NN	O	O
the	NN	O	O
inner	NN	O	O
retinal	NN	O	O
layers	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
visual	NN	O	B-Disease
deterioration	NN	O	I-Disease
in	NN	O	O
males	NN	O	O
.	NN	O	O

Many	NN	O	O
missense	NN	O	O
and	NN	O	O
protein	NN	O	O
-	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
causative	NN	O	O
gene	NN	O	O
RS1	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
identified	NN	O	O
and	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
inactivating	NN	O	O
.	NN	O	O

RS1	NN	O	O
encodes	NN	O	O
a	NN	O	O
224	NN	O	O
amino	NN	O	O
acid	NN	O	O
protein	NN	O	O
,	NN	O	O
retinoschisin	NN	O	O
,	NN	O	O
which	NN	O	O
contains	NN	O	O
a	NN	O	O
discoidin	NN	O	O
domain	NN	O	O
but	NN	O	O
is	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
antibody	NN	O	O
against	NN	O	O
a	NN	O	O
peptide	NN	O	O
from	NN	O	O
a	NN	O	O
unique	NN	O	O
region	NN	O	O
within	NN	O	O
retinoschisin	NN	O	O
,	NN	O	O
which	NN	O	O
detects	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
approximately	NN	O	O
28	NN	O	O
kDa	NN	O	O
in	NN	O	O
retinal	NN	O	O
samples	NN	O	O
reduced	NN	O	O
with	NN	O	O
dithiothreitol	NN	O	O
,	NN	O	O
but	NN	O	O
multimers	NN	O	O
sized	NN	O	O
>	NN	O	O
40	NN	O	O
kDa	NN	O	O
under	NN	O	O
non	NN	O	O
-	NN	O	O
reducing	NN	O	O
conditions	NN	O	O
.	NN	O	O

A	NN	O	O
screen	NN	O	O
of	NN	O	O
human	NN	O	O
tissues	NN	O	O
with	NN	O	O
this	NN	O	O
antibody	NN	O	O
reveals	NN	O	O
retinoschisin	NN	O	O
to	NN	O	O
be	NN	O	O
retina	NN	O	O
specific	NN	O	O
and	NN	O	O
the	NN	O	O
antibody	NN	O	O
detects	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
similar	NN	O	O
size	NN	O	O
in	NN	O	O
bovine	NN	O	O
and	NN	O	O
murine	NN	O	O
retinae	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
in	NN	O	O
the	NN	O	O
retina	NN	O	O
of	NN	O	O
both	NN	O	O
RS1	NN	O	O
mRNA	NN	O	O
(	NN	O	O
using	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
with	NN	O	O
riboprobes	NN	O	O
)	NN	O	O
and	NN	O	O
retinoschisin	NN	O	O
(	NN	O	O
using	NN	O	O
immunohistochemistry	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
antisense	NN	O	O
riboprobe	NN	O	O
detected	NN	O	O
RS1	NN	O	O
mRNA	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
photoreceptor	NN	O	O
layer	NN	O	O
but	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
was	NN	O	O
present	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
photoreceptors	NN	O	O
and	NN	O	O
within	NN	O	O
the	NN	O	O
inner	NN	O	O
portions	NN	O	O
of	NN	O	O
the	NN	O	O
retina	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
differentiated	NN	O	O
retinoblastoma	NN	O	B-Disease
cells	NN	O	O
(	NN	O	O
Weri	NN	O	O
-	NN	O	O
Rb1	NN	O	O
cells	NN	O	O
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
express	NN	O	O
RS1	NN	O	O
mRNA	NN	O	O
and	NN	O	O
to	NN	O	O
release	NN	O	O
retinoschisin	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
retinoschisin	NN	O	O
is	NN	O	O
released	NN	O	O
by	NN	O	O
photo	NN	O	O
-	NN	O	O
receptors	NN	O	O
and	NN	O	O
has	NN	O	O
functions	NN	O	O
within	NN	O	O
the	NN	O	O
inner	NN	O	O
retinal	NN	O	O
layers	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
retinoschisis	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
a	NN	O	O
putative	NN	O	O
secreted	NN	O	O
photoreceptor	NN	O	O
protein	NN	O	O
and	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
secreted	NN	O	O
photo	NN	O	O
-	NN	O	O
receptor	NN	O	O
protein	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
retinal	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Study	NN	O	O
of	NN	O	O
the	NN	O	O
voltage	NN	O	O
-	NN	O	O
gated	NN	O	O
sodium	NN	O	O
channel	NN	O	O
beta	NN	O	O
1	NN	O	O
subunit	NN	O	O
gene	NN	O	O
(	NN	O	O
SCN1B	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
benign	NN	O	B-Disease
familial	NN	O	I-Disease
infantile	NN	O	I-Disease
convulsions	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
BFIC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Benign	NN	O	B-Disease
familial	NN	O	I-Disease
infantile	NN	O	I-Disease
convulsions	NN	O	I-Disease
(	NN	O	O
BFIC	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
epilepsy	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
syndrome	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
described	NN	O	O
in	NN	O	O
Italian	NN	O	O
and	NN	O	O
French	NN	O	O
pedigrees	NN	O	O
.	NN	O	O

Patients	NN	O	O
present	NN	O	O
with	NN	O	O
partial	NN	O	O
,	NN	O	O
then	NN	O	O
generalized	NN	O	O
seizures	NN	O	O
,	NN	O	O
with	NN	O	O
onset	NN	O	O
at	NN	O	O
age	NN	O	O
three	NN	O	O
months	NN	O	O
.	NN	O	O

The	NN	O	O
seizures	NN	O	O
usually	NN	O	O
spontaneously	NN	O	O
cease	NN	O	O
after	NN	O	O
one	NN	O	O
year	NN	O	O
without	NN	O	O
treatment	NN	O	O
,	NN	O	O
leaving	NN	O	O
no	NN	O	O
neurological	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
mapped	NN	O	O
BFIC	NN	O	B-Disease
to	NN	O	O
chromosome	NN	O	O
19q	NN	O	O
in	NN	O	O
five	NN	O	O
Italian	NN	O	O
pedigrees	NN	O	O
.	NN	O	O

The	NN	O	O
sodium	NN	O	O
channel	NN	O	O
beta1	NN	O	O
subunit	NN	O	O
gene	NN	O	O
(	NN	O	O
SCN1B	NN	O	O
)	NN	O	O
maps	NN	O	O
to	NN	O	O
this	NN	O	O
candidate	NN	O	O
region	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
one	NN	O	O
Australian	NN	O	O
pedigree	NN	O	O
with	NN	O	O
generalized	NN	O	B-Disease
epilepsy	NN	O	I-Disease
and	NN	O	I-Disease
febrile	NN	O	I-Disease
seizures	NN	O	I-Disease
"	NN	O	I-Disease
plus	NN	O	I-Disease
"	NN	O	I-Disease
(	NN	O	O
GEFS	NN	O	O
+	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
SCN1B	NN	O	O
cosegregates	NN	O	O
with	NN	O	O
the	NN	O	O
GEFS	NN	O	O
+	NN	O	O
phenotype	NN	O	O
.	NN	O	O

BFIC	NN	O	B-Disease
and	NN	O	O
GEFS	NN	O	O
+	NN	O	O
have	NN	O	O
clinical	NN	O	O
features	NN	O	O
in	NN	O	O
common	NN	O	O
,	NN	O	O
therefore	NN	O	O
SCN1B	NN	O	O
is	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
BFIC	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
studied	NN	O	O
SCN1B	NN	O	O
exons	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
and	NN	O	O
5	NN	O	O
,	NN	O	O
using	NN	O	O
four	NN	O	O
SSCP	NN	O	O
methods	NN	O	O
in	NN	O	O
10	NN	O	O
Caucasian	NN	O	O
BFIC	NN	O	B-Disease
probands	NN	O	O
of	NN	O	O
Western	NN	O	O
Europe	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
no	NN	O	O
exon	NN	O	O
variants	NN	O	O
.	NN	O	O

One	NN	O	O
variant	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
intron	NN	O	O
5	NN	O	O
(	NN	O	O
IVS5	NN	O	O
-	NN	O	O
10C	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
segregate	NN	O	O
with	NN	O	O
BFIC	NN	O	B-Disease
and	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
9	NN	O	O
.	NN	O	O

2	NN	O	O
%	NN	O	O
controls	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
variant	NN	O	O
in	NN	O	O
intron	NN	O	O
5	NN	O	O
was	NN	O	O
identified	NN	O	O
(	NN	O	O
IVS5	NN	O	O
+	NN	O	O
30G	NN	O	O
>	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
rare	NN	O	O
,	NN	O	O
as	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
controls	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
segregating	NN	O	O
with	NN	O	O
the	NN	O	O
BFIC	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

Inactivation	NN	O	O
of	NN	O	O
germline	NN	O	O
mutant	NN	O	O
APC	NN	O	B-Disease
alleles	NN	O	O
by	NN	O	O
attenuated	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
:	NN	O	O
a	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
mechanism	NN	O	O
for	NN	O	O
attenuated	NN	O	B-Disease
familial	NN	O	I-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	I-Disease
APC	NN	O	I-Disease
)	NN	O	I-Disease
tumor	NN	O	I-Disease
-	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
result	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
FAP	NN	O	B-Disease
typically	NN	O	O
develop	NN	O	O
hundreds	NN	O	O
to	NN	O	O
thousands	NN	O	O
of	NN	O	O
benign	NN	O	B-Disease
colorectal	NN	O	I-Disease
tumors	NN	O	I-Disease
and	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
subset	NN	O	O
of	NN	O	O
germline	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
attenuated	NN	O	B-Disease
FAP	NN	O	I-Disease
(	NN	O	O
AFAP	NN	O	B-Disease
)	NN	O	O
phenotype	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
patients	NN	O	O
develop	NN	O	O
fewer	NN	O	O
tumors	NN	O	B-Disease
and	NN	O	O
develop	NN	O	O
them	NN	O	O
at	NN	O	O
an	NN	O	O
older	NN	O	O
age	NN	O	O
.	NN	O	O

Although	NN	O	O
a	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
locations	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
germline	NN	O	O
mutations	NN	O	O
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
AFAP	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
well	NN	O	O
documented	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
AFAP	NN	O	B-Disease
has	NN	O	O
not	NN	O	O
been	NN	O	O
well	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
AFAP	NN	O	B-Disease
in	NN	O	O
patients	NN	O	O
carrying	NN	O	O
a	NN	O	O
mutant	NN	O	O
APC	NN	O	B-Disease
allele	NN	O	O
(	NN	O	O
APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
)	NN	O	O
that	NN	O	O
has	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
region	NN	O	O
of	NN	O	O
exon	NN	O	O
9	NN	O	O
.	NN	O	O

APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
down	NN	O	O
-	NN	O	O
regulate	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
-	NN	O	O
regulated	NN	O	O
transcription	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
tumor	NN	O	B-Disease
-	NN	O	O
suppressor	NN	O	O
function	NN	O	O
of	NN	O	O
APC	NN	O	O
,	NN	O	O
as	NN	O	O
did	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	O
.	NN	O	O

Mutation	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	B-Disease
alleles	NN	O	O
were	NN	O	O
somatically	NN	O	O
mutated	NN	O	O
in	NN	O	O
most	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
from	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
4666insA	NN	O	O
,	NN	O	O
a	NN	O	O
common	NN	O	O
somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
in	NN	O	O
these	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
did	NN	O	O
not	NN	O	O
inactivate	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
carriers	NN	O	O
of	NN	O	O
APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
develop	NN	O	O
fewer	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
than	NN	O	O
do	NN	O	O
typical	NN	O	O
patients	NN	O	O
with	NN	O	O
FAP	NN	O	B-Disease
because	NN	O	O
somatic	NN	O	O
inactivation	NN	O	O
of	NN	O	O
both	NN	O	O
APC	NN	O	B-Disease
alleles	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
colorectal	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
patients	NN	O	O
develop	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
more	NN	O	O
frequently	NN	O	O
than	NN	O	O
does	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
because	NN	O	O
APC	NN	O	O
(	NN	O	O
AS9	NN	O	O
)	NN	O	O
is	NN	O	O
inactivated	NN	O	O
by	NN	O	O
mutations	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
inactivate	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
APC	NN	O	O
.	NN	O	O
.	NN	O	O

Iron	NN	O	O
-	NN	O	O
dependent	NN	O	O
self	NN	O	O
-	NN	O	O
assembly	NN	O	O
of	NN	O	O
recombinant	NN	O	O
yeast	NN	O	O
frataxin	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
.	NN	O	O

Frataxin	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
primary	NN	O	O
cause	NN	O	O
of	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
cardiodegenerative	NN	O	I-Disease
and	NN	O	I-Disease
neurodegenerative	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Frataxin	NN	O	O
is	NN	O	O
a	NN	O	O
nuclear	NN	O	O
-	NN	O	O
encoded	NN	O	O
mitochondrial	NN	O	O
protein	NN	O	O
that	NN	O	O
is	NN	O	O
widely	NN	O	O
conserved	NN	O	O
among	NN	O	O
eukaryotes	NN	O	O
.	NN	O	O

Genetic	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
yeast	NN	O	O
frataxin	NN	O	O
homologue	NN	O	O
(	NN	O	O
Yfh1p	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
mitochondrial	NN	O	B-Disease
iron	NN	O	I-Disease
accumulation	NN	O	I-Disease
and	NN	O	O
hypersensitivity	NN	O	B-Disease
to	NN	O	I-Disease
oxidative	NN	O	I-Disease
stress	NN	O	I-Disease
.	NN	O	O

Increased	NN	O	O
iron	NN	O	O
deposition	NN	O	O
and	NN	O	O
evidence	NN	O	O
of	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
have	NN	O	O
also	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
cardiac	NN	O	O
tissue	NN	O	O
and	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
FRDA	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
frataxin	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
mitochondrial	NN	O	O
iron	NN	O	O
homeostasis	NN	O	O
and	NN	O	O
protection	NN	O	O
from	NN	O	O
iron	NN	O	O
-	NN	O	O
induced	NN	O	O
formation	NN	O	O
of	NN	O	O
free	NN	O	O
radicals	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
mechanism	NN	O	O
of	NN	O	O
frataxin	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
is	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
expressed	NN	O	O
the	NN	O	O
mature	NN	O	O
form	NN	O	O
of	NN	O	O
Yfh1p	NN	O	O
(	NN	O	O
mYfh1p	NN	O	O
)	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
and	NN	O	O
have	NN	O	O
analyzed	NN	O	O
its	NN	O	O
function	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Isolated	NN	O	O
mYfh1p	NN	O	O
is	NN	O	O
a	NN	O	O
soluble	NN	O	O
monomer	NN	O	O
(	NN	O	O
13	NN	O	O
,	NN	O	O
783	NN	O	O
Da	NN	O	O
)	NN	O	O
that	NN	O	O
contains	NN	O	O
no	NN	O	O
iron	NN	O	O
and	NN	O	O
shows	NN	O	O
no	NN	O	O
significant	NN	O	O
tendency	NN	O	O
to	NN	O	O
self	NN	O	O
-	NN	O	O
associate	NN	O	O
.	NN	O	O

Aerobic	NN	O	O
addition	NN	O	O
of	NN	O	O
ferrous	NN	O	O
iron	NN	O	O
to	NN	O	O
mYfh1p	NN	O	O
results	NN	O	O
in	NN	O	O
assembly	NN	O	O
of	NN	O	O
regular	NN	O	O
spherical	NN	O	O
multimers	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
approximately	NN	O	O
1	NN	O	O
.	NN	O	O

1	NN	O	O
MDa	NN	O	O
(	NN	O	O
megadaltons	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
diameter	NN	O	O
of	NN	O	O
13	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
2	NN	O	O
nm	NN	O	O
.	NN	O	O

Each	NN	O	O
multimer	NN	O	O
consists	NN	O	O
of	NN	O	O
approximately	NN	O	O
60	NN	O	O
subunits	NN	O	O
and	NN	O	O
can	NN	O	O
sequester	NN	O	O
>	NN	O	O
3	NN	O	O
,	NN	O	O
000	NN	O	O
atoms	NN	O	O
of	NN	O	O
iron	NN	O	O
.	NN	O	O

Titration	NN	O	O
of	NN	O	O
mYfh1p	NN	O	O
with	NN	O	O
increasing	NN	O	O
iron	NN	O	O
concentrations	NN	O	O
supports	NN	O	O
a	NN	O	O
stepwise	NN	O	O
mechanism	NN	O	O
of	NN	O	O
multimer	NN	O	O
assembly	NN	O	O
.	NN	O	O

Sequential	NN	O	O
addition	NN	O	O
of	NN	O	O
an	NN	O	O
iron	NN	O	O
chelator	NN	O	O
and	NN	O	O
a	NN	O	O
reducing	NN	O	O
agent	NN	O	O
results	NN	O	O
in	NN	O	O
quantitative	NN	O	O
iron	NN	O	O
release	NN	O	O
with	NN	O	O
concomitant	NN	O	O
disassembly	NN	O	O
of	NN	O	O
the	NN	O	O
multimer	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
mYfh1p	NN	O	O
sequesters	NN	O	O
iron	NN	O	O
in	NN	O	O
an	NN	O	O
available	NN	O	O
form	NN	O	O
.	NN	O	O

In	NN	O	O
yeast	NN	O	O
mitochondria	NN	O	O
,	NN	O	O
native	NN	O	O
mYfh1p	NN	O	O
exists	NN	O	O
as	NN	O	O
monomer	NN	O	O
and	NN	O	O
a	NN	O	O
higher	NN	O	O
-	NN	O	O
order	NN	O	O
species	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
>	NN	O	O
600	NN	O	O
,	NN	O	O
000	NN	O	O
.	NN	O	O

After	NN	O	O
addition	NN	O	O
of	NN	O	O
(	NN	O	O
55	NN	O	O
)	NN	O	O
Fe	NN	O	O
to	NN	O	O
the	NN	O	O
medium	NN	O	O
,	NN	O	O
immunoprecipitates	NN	O	O
of	NN	O	O
this	NN	O	O
species	NN	O	O
contain	NN	O	O
>	NN	O	O
16	NN	O	O
atoms	NN	O	O
of	NN	O	O
(	NN	O	O
55	NN	O	O
)	NN	O	O
Fe	NN	O	O
per	NN	O	O
molecule	NN	O	O
of	NN	O	O
mYfh1p	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
iron	NN	O	O
-	NN	O	O
dependent	NN	O	O
self	NN	O	O
-	NN	O	O
assembly	NN	O	O
of	NN	O	O
recombinant	NN	O	O
mYfh1p	NN	O	O
reflects	NN	O	O
a	NN	O	O
physiological	NN	O	O
role	NN	O	O
for	NN	O	O
frataxin	NN	O	O
in	NN	O	O
mitochondrial	NN	O	O
iron	NN	O	O
sequestration	NN	O	O
and	NN	O	O
bioavailability	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
pleckstrin	NN	O	O
homology	NN	O	O
(	NN	O	O
PH	NN	O	O
)	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
in	NN	O	O
an	NN	O	O
Italian	NN	O	O
family	NN	O	O
with	NN	O	O
faciogenital	NN	O	B-Disease
dysplasia	NN	O	I-Disease
(	NN	O	O
Aarskog	NN	O	B-Disease
-	NN	O	I-Disease
Scott	NN	O	I-Disease
syndrome	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

Aarskog	NN	O	B-Disease
-	NN	O	I-Disease
Scott	NN	O	I-Disease
Syndrome	NN	O	I-Disease
(	NN	O	O
AAS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disorder	NN	O	I-Disease
characterised	NN	O	O
by	NN	O	O
short	NN	O	B-Disease
stature	NN	O	I-Disease
and	NN	O	O
multiple	NN	O	O
facial	NN	O	B-Disease
,	NN	O	I-Disease
limb	NN	O	I-Disease
and	NN	O	I-Disease
genital	NN	O	I-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
gene	NN	O	O
,	NN	O	O
FGD1	NN	O	O
,	NN	O	O
altered	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
AAS	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
and	NN	O	O
found	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
homology	NN	O	O
to	NN	O	O
Rho	NN	O	O
/	NN	O	O
Rac	NN	O	O
guanine	NN	O	O
nucleotide	NN	O	O
exchange	NN	O	O
factors	NN	O	O
(	NN	O	O
Rho	NN	O	O
/	NN	O	O
Rac	NN	O	O
GEF	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
since	NN	O	O
this	NN	O	O
original	NN	O	O
report	NN	O	O
on	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
mutated	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
in	NN	O	O
an	NN	O	O
AAS	NN	O	B-Disease
patient	NN	O	O
,	NN	O	O
no	NN	O	O
additional	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
13	NN	O	O
independent	NN	O	O
patients	NN	O	O
with	NN	O	O
clinical	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
AAS	NN	O	B-Disease
.	NN	O	O

One	NN	O	O
patient	NN	O	O
presented	NN	O	O
a	NN	O	O
mutation	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
nucleotide	NN	O	O
change	NN	O	O
in	NN	O	O
exon	NN	O	O
10	NN	O	O
of	NN	O	O
the	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
(	NN	O	O
G2559	NN	O	O
>	NN	O	O
A	NN	O	O
)	NN	O	O
substituting	NN	O	O
a	NN	O	O
Gln	NN	O	O
for	NN	O	O
Arg	NN	O	O
in	NN	O	O
position	NN	O	O
610	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
segregate	NN	O	O
with	NN	O	O
the	NN	O	O
AAS	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
carrier	NN	O	O
females	NN	O	O
in	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
Arg	NN	O	O
-	NN	O	O
610	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
pleckstrin	NN	O	O
homology	NN	O	O
(	NN	O	O
PH	NN	O	O
)	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
and	NN	O	O
it	NN	O	O
corresponds	NN	O	O
to	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
residue	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
involved	NN	O	O
in	NN	O	O
InsP	NN	O	O
binding	NN	O	O
in	NN	O	O
PH	NN	O	O
domains	NN	O	O
of	NN	O	O
other	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
residue	NN	O	O
is	NN	O	O
often	NN	O	O
mutated	NN	O	O
in	NN	O	O
the	NN	O	O
Brutons	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
(	NN	O	O
Btk	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
agammaglobulinemia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Arg610Gln	NN	O	O
mutation	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
case	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
PH	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
FGD1	NN	O	O
gene	NN	O	O
and	NN	O	O
additional	NN	O	O
evidence	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
PH	NN	O	O
domains	NN	O	O
can	NN	O	O
be	NN	O	O
associated	NN	O	O
to	NN	O	O
human	NN	O	O
diseases	NN	O	O
.	NN	O	O
.	NN	O	O

Amino	NN	O	O
-	NN	O	O
terminal	NN	O	O
fragments	NN	O	O
of	NN	O	O
mutant	NN	O	O
huntingtin	NN	O	O
show	NN	O	O
selective	NN	O	O
accumulation	NN	O	O
in	NN	O	O
striatal	NN	O	O
neurons	NN	O	O
and	NN	O	O
synaptic	NN	O	O
toxicity	NN	O	O
.	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
glutamine	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
amino	NN	O	O
-	NN	O	O
terminal	NN	O	O
region	NN	O	O
of	NN	O	O
huntingtin	NN	O	O
.	NN	O	O

Despite	NN	O	O
its	NN	O	O
widespread	NN	O	O
expression	NN	O	O
,	NN	O	O
mutant	NN	O	O
huntingtin	NN	O	O
induces	NN	O	O
selective	NN	O	O
neuronal	NN	O	O
loss	NN	O	O
in	NN	O	O
striatal	NN	O	O
neurons	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
mutant	NN	O	O
mice	NN	O	O
expressing	NN	O	O
HD	NN	O	B-Disease
repeats	NN	O	O
,	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
aggregation	NN	O	O
of	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
huntingtin	NN	O	O
fragments	NN	O	O
preferentially	NN	O	O
occur	NN	O	O
in	NN	O	O
HD	NN	O	B-Disease
-	NN	O	O
affected	NN	O	O
neurons	NN	O	O
and	NN	O	O
their	NN	O	O
processes	NN	O	O
and	NN	O	O
axonal	NN	O	O
terminals	NN	O	O
.	NN	O	O

N	NN	O	O
-	NN	O	O
terminal	NN	O	O
fragments	NN	O	O
of	NN	O	O
mutant	NN	O	O
huntingtin	NN	O	O
form	NN	O	O
aggregates	NN	O	O
and	NN	O	O
induce	NN	O	O
neuritic	NN	O	B-Disease
degeneration	NN	O	I-Disease
in	NN	O	O
cultured	NN	O	O
striatal	NN	O	O
neurons	NN	O	O
.	NN	O	O

N	NN	O	O
-	NN	O	O
terminal	NN	O	O
mutant	NN	O	O
huntingtin	NN	O	O
also	NN	O	O
binds	NN	O	O
to	NN	O	O
synaptic	NN	O	O
vesicles	NN	O	O
and	NN	O	O
inhibits	NN	O	O
their	NN	O	O
glutamate	NN	O	O
uptake	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
specific	NN	O	O
processing	NN	O	O
and	NN	O	O
accumulation	NN	O	O
of	NN	O	O
toxic	NN	O	O
fragments	NN	O	O
of	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
huntingtin	NN	O	O
in	NN	O	O
HD	NN	O	B-Disease
-	NN	O	O
affected	NN	O	O
striatal	NN	O	O
neurons	NN	O	O
,	NN	O	O
especially	NN	O	O
in	NN	O	O
their	NN	O	O
neuronal	NN	O	O
processes	NN	O	O
and	NN	O	O
axonal	NN	O	O
terminals	NN	O	O
,	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
selective	NN	O	O
neuropathology	NN	O	O
of	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
human	NN	O	O
SWI	NN	O	O
/	NN	O	O
SNF	NN	O	O
-	NN	O	O
related	NN	O	O
complex	NN	O	O
:	NN	O	O
linking	NN	O	O
chromatin	NN	O	O
remodeling	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
predispose	NN	O	O
individuals	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

Using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
affinity	NN	O	O
-	NN	O	O
and	NN	O	O
conventional	NN	O	O
chromatographic	NN	O	O
techniques	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
a	NN	O	O
predominant	NN	O	O
form	NN	O	O
of	NN	O	O
a	NN	O	O
multiprotein	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
containing	NN	O	O
complex	NN	O	O
from	NN	O	O
human	NN	O	O
cells	NN	O	O
displaying	NN	O	O
chromatin	NN	O	O
-	NN	O	O
remodeling	NN	O	O
activity	NN	O	O
.	NN	O	O

Mass	NN	O	O
spectrometric	NN	O	O
sequencing	NN	O	O
of	NN	O	O
components	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
indicated	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
SWI	NN	O	O
/	NN	O	O
SNF	NN	O	O
-	NN	O	O
related	NN	O	O
complex	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
can	NN	O	O
directly	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
BRG1	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
SWI	NN	O	O
/	NN	O	O
SNF	NN	O	O
complex	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
p53	NN	O	O
-	NN	O	O
mediated	NN	O	O
stimulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
BRCA1	NN	O	O
was	NN	O	O
completely	NN	O	O
abrogated	NN	O	O
by	NN	O	O
either	NN	O	O
a	NN	O	O
dominant	NN	O	O
-	NN	O	O
negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
BRG1	NN	O	O
or	NN	O	O
the	NN	O	O
cancer	NN	O	B-Disease
-	NN	O	O
causing	NN	O	O
deletion	NN	O	O
in	NN	O	O
exon	NN	O	O
11	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
reveal	NN	O	O
a	NN	O	O
direct	NN	O	O
function	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
through	NN	O	O
modulation	NN	O	O
of	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
.	NN	O	O
.	NN	O	O

Asef	NN	O	O
,	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
APC	NN	O	O
and	NN	O	O
G	NN	O	O
-	NN	O	O
protein	NN	O	O
signaling	NN	O	O
.	NN	O	O

The	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
APC	NN	O	O
)	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
and	NN	O	O
in	NN	O	O
sporadic	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

Here	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
through	NN	O	O
its	NN	O	O
armadillo	NN	O	O
repeat	NN	O	O
domain	NN	O	O
to	NN	O	O
a	NN	O	O
Rac	NN	O	O
-	NN	O	O
specific	NN	O	O
guanine	NN	O	O
nucleotide	NN	O	O
exchange	NN	O	O
factor	NN	O	O
(	NN	O	O
GEF	NN	O	O
)	NN	O	O
,	NN	O	O
termed	NN	O	O
Asef	NN	O	O
.	NN	O	O

Endogenous	NN	O	O
APC	NN	O	O
colocalized	NN	O	O
with	NN	O	O
Asef	NN	O	O
in	NN	O	O
mouse	NN	O	O
colon	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
and	NN	O	O
neuronal	NN	O	O
cells	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
APC	NN	O	O
enhanced	NN	O	O
the	NN	O	O
GEF	NN	O	O
activity	NN	O	O
of	NN	O	O
Asef	NN	O	O
and	NN	O	O
stimulated	NN	O	O
Asef	NN	O	O
-	NN	O	O
mediated	NN	O	O
cell	NN	O	O
flattening	NN	O	O
,	NN	O	O
membrane	NN	O	O
ruffling	NN	O	O
,	NN	O	O
and	NN	O	O
lamellipodia	NN	O	O
formation	NN	O	O
in	NN	O	O
MDCK	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
APC	NN	O	O
-	NN	O	O
Asef	NN	O	O
complex	NN	O	O
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
actin	NN	O	O
cytoskeletal	NN	O	O
network	NN	O	O
,	NN	O	O
cell	NN	O	O
morphology	NN	O	O
and	NN	O	O
migration	NN	O	O
,	NN	O	O
and	NN	O	O
neuronal	NN	O	O
function	NN	O	O
.	NN	O	O
.	NN	O	O

Function	NN	O	O
of	NN	O	O
an	NN	O	O
axonal	NN	O	O
chemoattractant	NN	O	O
modulated	NN	O	O
by	NN	O	O
metalloprotease	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
axonal	NN	O	O
chemoattractant	NN	O	O
netrin	NN	O	O
-	NN	O	O
1	NN	O	O
guides	NN	O	O
spinal	NN	O	O
commissural	NN	O	O
axons	NN	O	O
by	NN	O	O
activating	NN	O	O
its	NN	O	O
receptor	NN	O	O
DCC	NN	O	O
(	NN	O	O
Deleted	NN	O	O
in	NN	O	O
Colorectal	NN	O	B-Disease
Cancer	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
chemical	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
metalloproteases	NN	O	O
potentiate	NN	O	O
netrin	NN	O	O
-	NN	O	O
mediated	NN	O	O
axon	NN	O	O
outgrowth	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
DCC	NN	O	O
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
metalloprotease	NN	O	O
-	NN	O	O
dependent	NN	O	O
ectodomain	NN	O	O
shedding	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
inhibitors	NN	O	O
block	NN	O	O
proteolytic	NN	O	O
processing	NN	O	O
of	NN	O	O
DCC	NN	O	O
and	NN	O	O
cause	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
DCC	NN	O	O
protein	NN	O	O
levels	NN	O	O
on	NN	O	O
axons	NN	O	O
within	NN	O	O
spinal	NN	O	O
cord	NN	O	O
explants	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
potentiation	NN	O	O
of	NN	O	O
netrin	NN	O	O
activity	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
may	NN	O	O
result	NN	O	O
from	NN	O	O
stabilization	NN	O	O
of	NN	O	O
DCC	NN	O	O
on	NN	O	O
the	NN	O	O
axons	NN	O	O
,	NN	O	O
and	NN	O	O
proteolytic	NN	O	O
activity	NN	O	O
may	NN	O	O
regulate	NN	O	O
axon	NN	O	O
migration	NN	O	O
by	NN	O	O
controlling	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
functional	NN	O	O
extracellular	NN	O	O
axon	NN	O	O
guidance	NN	O	O
receptors	NN	O	O
.	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
an	NN	O	O
expanded	NN	O	O
CUG	NN	O	O
repeat	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
in	NN	O	O
adult	NN	O	O
humans	NN	O	O
,	NN	O	O
results	NN	O	O
from	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
DMPK	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
DMPK	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
(	NN	O	O
mRNA	NN	O	O
)	NN	O	O
contains	NN	O	O
an	NN	O	O
expanded	NN	O	O
CUG	NN	O	O
repeat	NN	O	O
and	NN	O	O
is	NN	O	O
retained	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
expressed	NN	O	O
an	NN	O	O
untranslated	NN	O	O
CUG	NN	O	O
repeat	NN	O	O
in	NN	O	O
an	NN	O	O
unrelated	NN	O	O
mRNA	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Mice	NN	O	O
that	NN	O	O
expressed	NN	O	O
expanded	NN	O	O
CUG	NN	O	O
repeats	NN	O	O
developed	NN	O	O
myotonia	NN	O	B-Disease
and	NN	O	O
myopathy	NN	O	B-Disease
,	NN	O	O
whereas	NN	O	O
mice	NN	O	O
expressing	NN	O	O
a	NN	O	O
nonexpanded	NN	O	O
repeat	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
transcripts	NN	O	O
with	NN	O	O
expanded	NN	O	O
CUG	NN	O	O
repeats	NN	O	O
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
generate	NN	O	O
a	NN	O	O
DM	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
supports	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
RNA	NN	O	O
gain	NN	O	O
of	NN	O	O
function	NN	O	O
in	NN	O	O
disease	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O
.	NN	O	O

Genomic	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
tumor	NN	O	I-Disease
-	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	I-Disease
APC	NN	O	I-Disease
)	NN	O	I-Disease
tumor	NN	O	I-Disease
-	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
colorectal	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Almost	NN	O	O
all	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
are	NN	O	O
single	NN	O	O
-	NN	O	O
nucleotide	NN	O	O
alterations	NN	O	O
,	NN	O	O
small	NN	O	O
insertions	NN	O	O
,	NN	O	O
or	NN	O	O
small	NN	O	O
deletions	NN	O	O
that	NN	O	O
would	NN	O	O
truncate	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

No	NN	O	O
well	NN	O	O
-	NN	O	O
characterized	NN	O	O
intragenic	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
APC	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
this	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
.	NN	O	O

We	NN	O	O
screened	NN	O	O
49	NN	O	O
potential	NN	O	O
FAP	NN	O	B-Disease
families	NN	O	O
and	NN	O	O
identified	NN	O	O
26	NN	O	O
different	NN	O	O
germline	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
30	NN	O	O
families	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
were	NN	O	O
genomic	NN	O	O
rearrangements	NN	O	O
resulting	NN	O	O
from	NN	O	O
homologous	NN	O	O
and	NN	O	O
nonhomologous	NN	O	O
recombinations	NN	O	O
mediated	NN	O	O
by	NN	O	O
Alu	NN	O	O
elements	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
four	NN	O	O
rearrangements	NN	O	O
were	NN	O	O
complex	NN	O	O
,	NN	O	O
involving	NN	O	O
deletion	NN	O	O
and	NN	O	O
insertion	NN	O	O
of	NN	O	O
nucleotides	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
four	NN	O	O
rearrangements	NN	O	O
,	NN	O	O
one	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
exons	NN	O	O
11	NN	O	O
and	NN	O	O
12	NN	O	O
and	NN	O	O
two	NN	O	O
others	NN	O	O
resulted	NN	O	O
in	NN	O	O
either	NN	O	O
complete	NN	O	O
or	NN	O	O
partial	NN	O	O
deletion	NN	O	O
of	NN	O	O
exon	NN	O	O
14	NN	O	O
.	NN	O	O

The	NN	O	O
fourth	NN	O	O
rearrangement	NN	O	O
grossly	NN	O	O
altered	NN	O	O
the	NN	O	O
sequence	NN	O	O
within	NN	O	O
intron	NN	O	O
14	NN	O	O
.	NN	O	O

Although	NN	O	O
this	NN	O	O
rearrangement	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
any	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
APC	NN	O	O
at	NN	O	O
the	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
level	NN	O	O
,	NN	O	O
it	NN	O	O
caused	NN	O	O
inappropriate	NN	O	O
splicing	NN	O	O
of	NN	O	O
exon	NN	O	O
14	NN	O	O
.	NN	O	O

These	NN	O	O
rearrangements	NN	O	O
were	NN	O	O
initially	NN	O	O
revealed	NN	O	O
by	NN	O	O
analyzing	NN	O	O
cDNAs	NN	O	O
and	NN	O	O
could	NN	O	O
not	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
by	NN	O	O
using	NN	O	O
mutation	NN	O	O
detection	NN	O	O
methods	NN	O	O
that	NN	O	O
screened	NN	O	O
each	NN	O	O
exon	NN	O	O
individually	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
rearrangement	NN	O	O
that	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
any	NN	O	O
coding	NN	O	O
exons	NN	O	O
yet	NN	O	O
affected	NN	O	O
the	NN	O	O
splicing	NN	O	O
further	NN	O	O
underscores	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
using	NN	O	O
cDNA	NN	O	O
for	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
four	NN	O	O
genomic	NN	O	O
rearrangements	NN	O	O
among	NN	O	O
30	NN	O	O
mutations	NN	O	O
suggests	NN	O	O
that	NN	O	O
genomic	NN	O	O
rearrangements	NN	O	O
are	NN	O	O
frequent	NN	O	O
germline	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
missense	NN	O	O
mutation	NN	O	O
and	NN	O	O
second	NN	O	O
recurrent	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
CACNA1A	NN	O	O
gene	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
EA	NN	O	B-Disease
-	NN	O	I-Disease
2	NN	O	I-Disease
and	NN	O	O
FHM	NN	O	B-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
specific	NN	O	O
P	NN	O	O
/	NN	O	O
Q	NN	O	O
type	NN	O	O
Ca2	NN	O	O
+	NN	O	O
channel	NN	O	O
alpha1	NN	O	O
subunit	NN	O	O
gene	NN	O	O
,	NN	O	O
CACNA1A	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
three	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
disorders	NN	O	O
,	NN	O	O
viz	NN	O	O
.	NN	O	O
episodic	NN	O	B-Disease
ataxia	NN	O	I-Disease
type	NN	O	I-Disease
2	NN	O	I-Disease
(	NN	O	O
EA	NN	O	B-Disease
-	NN	O	I-Disease
2	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
familial	NN	O	B-Disease
hemiplegic	NN	O	I-Disease
migraine	NN	O	I-Disease
(	NN	O	O
FHM	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
spinocerebellar	NN	O	B-Disease
ataxia	NN	O	I-Disease
6	NN	O	I-Disease
(	NN	O	O
SCA6	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

For	NN	O	O
individuals	NN	O	O
with	NN	O	O
EA	NN	O	B-Disease
-	NN	O	I-Disease
2	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
mutations	NN	O	O
described	NN	O	O
thus	NN	O	O
far	NN	O	O
are	NN	O	O
presumed	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
truncated	NN	O	O
protein	NN	O	O
product	NN	O	O
.	NN	O	O

Several	NN	O	O
different	NN	O	O
missense	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
FHM	NN	O	B-Disease
.	NN	O	O

At	NN	O	O
least	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
on	NN	O	O
two	NN	O	O
different	NN	O	O
chromosome	NN	O	O
19p13	NN	O	O
haplotypes	NN	O	O
and	NN	O	O
thus	NN	O	O
represent	NN	O	O
recurrent	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
screened	NN	O	O
several	NN	O	O
individuals	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
all	NN	O	O
47	NN	O	O
exons	NN	O	O
in	NN	O	O
the	NN	O	O
CACNA1A	NN	O	O
gene	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
analysis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
characterised	NN	O	O
a	NN	O	O
novel	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
G5260A	NN	O	O
,	NN	O	O
in	NN	O	O
exon	NN	O	O
32	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
segregating	NN	O	O
for	NN	O	O
EA	NN	O	B-Disease
-	NN	O	I-Disease
2	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
consequence	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
is	NN	O	O
an	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
at	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
position	NN	O	O
within	NN	O	O
the	NN	O	O
CACNA1A	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
point	NN	O	O
mutation	NN	O	O
not	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
proposed	NN	O	O
truncated	NN	O	O
protein	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
this	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
member	NN	O	O
with	NN	O	O
mild	NN	O	O
clinical	NN	O	O
signs	NN	O	O
including	NN	O	O
only	NN	O	O
migraine	NN	O	B-Disease
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
a	NN	O	O
second	NN	O	O
previously	NN	O	O
identified	NN	O	O
recurrent	NN	O	O
muta	NN	O	O
tion	NN	O	O
,	NN	O	O
C2272T	NN	O	O
,	NN	O	O
in	NN	O	O
exon	NN	O	O
16	NN	O	O
has	NN	O	O
been	NN	O	O
discovered	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
FHM	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Analbuminemia	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
neonate	NN	O	O
.	NN	O	O

A	NN	O	O
small	NN	O	O
-	NN	O	O
for	NN	O	O
-	NN	O	O
gestational	NN	O	O
-	NN	O	O
age	NN	O	O
infant	NN	O	O
,	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
analbuminemia	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
neonatal	NN	O	O
period	NN	O	O
,	NN	O	O
is	NN	O	O
reported	NN	O	O
and	NN	O	O
the	NN	O	O
twelve	NN	O	O
cases	NN	O	O
recorded	NN	O	O
in	NN	O	O
the	NN	O	O
world	NN	O	O
literature	NN	O	O
are	NN	O	O
reviewed	NN	O	O
.	NN	O	O

Patients	NN	O	O
lacking	NN	O	O
this	NN	O	O
serum	NN	O	O
protein	NN	O	O
are	NN	O	O
essentially	NN	O	O
asymptomatic	NN	O	O
,	NN	O	O
apart	NN	O	O
from	NN	O	O
minimal	NN	O	O
ankle	NN	O	B-Disease
edema	NN	O	I-Disease
and	NN	O	O
ease	NN	O	B-Disease
of	NN	O	I-Disease
fatigue	NN	O	I-Disease
.	NN	O	O

Apparent	NN	O	O
compensatory	NN	O	O
mechanisms	NN	O	O
which	NN	O	O
come	NN	O	O
into	NN	O	O
play	NN	O	O
when	NN	O	O
serum	NN	O	O
albumin	NN	O	O
is	NN	O	O
low	NN	O	O
include	NN	O	O
prolonged	NN	O	O
half	NN	O	O
-	NN	O	O
life	NN	O	O
of	NN	O	O
albumin	NN	O	O
and	NN	O	O
transferrin	NN	O	O
,	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
serum	NN	O	O
globulins	NN	O	O
,	NN	O	O
beta	NN	O	O
lipoprotein	NN	O	O
,	NN	O	O
and	NN	O	O
glycoproteins	NN	O	O
,	NN	O	O
arterial	NN	O	B-Disease
hypotension	NN	O	I-Disease
with	NN	O	O
reduced	NN	O	O
capillary	NN	O	O
hydrostatic	NN	O	O
pressure	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
respond	NN	O	O
with	NN	O	O
rapid	NN	O	O
sodium	NN	O	O
and	NN	O	O
chloride	NN	O	O
diuresis	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
small	NN	O	O
volume	NN	O	O
changes	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
plasma	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
an	NN	O	O
investigation	NN	O	O
not	NN	O	O
previously	NN	O	O
reported	NN	O	O
,	NN	O	O
revealed	NN	O	O
an	NN	O	O
extremely	NN	O	O
low	NN	O	O
plasma	NN	O	O
tryptophan	NN	O	O
level	NN	O	O
,	NN	O	O
a	NN	O	O
finding	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
tryptophan	NN	O	O
in	NN	O	O
albumin	NN	O	O
synthesis	NN	O	O
.	NN	O	O
.	NN	O	O

Histidinemia	NN	O	B-Disease
.	NN	O	O

Classical	NN	O	O
and	NN	O	O
atypical	NN	O	O
form	NN	O	O
in	NN	O	O
siblings	NN	O	O
.	NN	O	O

Two	NN	O	O
brothers	NN	O	O
,	NN	O	O
6	NN	O	O
and	NN	O	O
13	NN	O	O
years	NN	O	O
old	NN	O	O
,	NN	O	O
had	NN	O	O
histidinemia	NN	O	B-Disease
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
observations	NN	O	O
,	NN	O	O
the	NN	O	O
younger	NN	O	O
boy	NN	O	O
was	NN	O	O
considered	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
classical	NN	O	O
type	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
older	NN	O	O
boy	NN	O	O
had	NN	O	O
an	NN	O	O
atypical	NN	O	O
form	NN	O	O
characterized	NN	O	O
by	NN	O	O
partial	NN	O	O
impairment	NN	O	O
of	NN	O	O
the	NN	O	O
skin	NN	O	O
histidase	NN	O	O
activity	NN	O	O
and	NN	O	O
a	NN	O	O
moderately	NN	O	O
prolonged	NN	O	O
half	NN	O	O
-	NN	O	O
life	NN	O	O
of	NN	O	O
blood	NN	O	O
histidine	NN	O	O
.	NN	O	O

The	NN	O	O
mother	NN	O	O
is	NN	O	O
a	NN	O	O
heterozygous	NN	O	O
carrier	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
father	NN	O	O
and	NN	O	O
sister	NN	O	O
seem	NN	O	O
to	NN	O	O
be	NN	O	O
normal	NN	O	O
.	NN	O	O
.	NN	O	O

Carrier	NN	O	O
detection	NN	O	O
of	NN	O	O
pyruvate	NN	O	B-Disease
carboxylase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
fibroblasts	NN	O	O
and	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Pyruvate	NN	O	O
carboxylase	NN	O	O
(	NN	O	O
E	NN	O	O
.	NN	O	O
C	NN	O	O
.	NN	O	O
6	NN	O	O
.	NN	O	O
4	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
activity	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
the	NN	O	O
circulating	NN	O	O
peripheral	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
hepatic	NN	O	O
,	NN	O	O
cerebral	NN	O	O
,	NN	O	O
renal	NN	O	O
cortical	NN	O	O
,	NN	O	O
leukocyte	NN	O	O
,	NN	O	O
and	NN	O	O
fibroblast	NN	O	O
pyruvate	NN	O	B-Disease
carboxylase	NN	O	I-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
PC	NN	O	B-Disease
Portland	NN	O	I-Disease
deficiency	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

Lymphocyte	NN	O	O
activities	NN	O	O
were	NN	O	O
mother	NN	O	O
,	NN	O	O
33	NN	O	O
-	NN	O	O
-	NN	O	O
39	NN	O	O
%	NN	O	O
;	NN	O	O
father	NN	O	O
,	NN	O	O
11	NN	O	O
-	NN	O	O
-	NN	O	O
29	NN	O	O
%	NN	O	O
;	NN	O	O
brother	NN	O	O
,	NN	O	O
82	NN	O	O
-	NN	O	O
-	NN	O	O
103	NN	O	O
%	NN	O	O
;	NN	O	O
and	NN	O	O
sister	NN	O	O
,	NN	O	O
38	NN	O	O
-	NN	O	O
-	NN	O	O
48	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
lowest	NN	O	O
normal	NN	O	O
.	NN	O	O

Fibroblasts	NN	O	O
from	NN	O	O
the	NN	O	O
patients	NN	O	O
mother	NN	O	O
and	NN	O	O
father	NN	O	O
had	NN	O	O
42	NN	O	O
and	NN	O	O
34	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lowest	NN	O	O
normal	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
disease	NN	O	O
is	NN	O	O
inherited	NN	O	O
in	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
manner	NN	O	O
and	NN	O	O
that	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
fibroblasts	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
carriers	NN	O	O
.	NN	O	O

Neither	NN	O	O
pyruvate	NN	O	O
carboxylase	NN	O	O
nor	NN	O	O
mitochondrial	NN	O	O
PEPCK	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
a	NN	O	O
21	NN	O	O
-	NN	O	O
hr	NN	O	O
fast	NN	O	O
.	NN	O	O

Glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variants	NN	O	O
from	NN	O	O
Italian	NN	O	O
subjects	NN	O	O
associated	NN	O	O
with	NN	O	O
severe	NN	O	B-Disease
neonatal	NN	O	I-Disease
jaundice	NN	O	I-Disease
.	NN	O	O

Screening	NN	O	O
for	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
carried	NN	O	O
out	NN	O	O
at	NN	O	O
the	NN	O	O
Maternity	NN	O	O
Division	NN	O	O
of	NN	O	O
the	NN	O	O
Galliera	NN	O	O
Hospital	NN	O	O
in	NN	O	O
Genoa	NN	O	O
,	NN	O	O
Italy	NN	O	O
.	NN	O	O

Two	NN	O	O
groups	NN	O	O
of	NN	O	O
subjects	NN	O	O
with	NN	O	O
hyperbilirubinaemia	NN	O	B-Disease
of	NN	O	O
non	NN	O	O
-	NN	O	O
immunological	NN	O	O
origin	NN	O	O
were	NN	O	O
examined	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
302	NN	O	O
newborn	NN	O	O
babies	NN	O	O
of	NN	O	O
Sardinian	NN	O	O
extraction	NN	O	O
(	NN	O	O
on	NN	O	O
cord	NN	O	O
blood	NN	O	O
)	NN	O	O
and	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
201	NN	O	O
newborn	NN	O	O
babies	NN	O	O
of	NN	O	O
south	NN	O	O
Italian	NN	O	O
ancestry	NN	O	O
(	NN	O	O
on	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
)	NN	O	O
.	NN	O	O

Among	NN	O	O
503	NN	O	O
subjects	NN	O	O
,	NN	O	O
43	NN	O	O
showed	NN	O	O
an	NN	O	O
enzyme	NN	O	O
deficiency	NN	O	O
;	NN	O	O
in	NN	O	O
39	NN	O	O
the	NN	O	O
defect	NN	O	O
was	NN	O	O
of	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
type	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
case	NN	O	O
,	NN	O	O
previously	NN	O	O
described	NN	O	O
,	NN	O	O
the	NN	O	O
enzyme	NN	O	O
was	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	O
-	NN	O	O
type	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
remaining	NN	O	O
cases	NN	O	O
three	NN	O	O
different	NN	O	O
variants	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
work	NN	O	O
these	NN	O	O
three	NN	O	O
cases	NN	O	O
,	NN	O	O
each	NN	O	O
with	NN	O	O
severe	NN	O	B-Disease
neonatal	NN	O	I-Disease
jaundice	NN	O	I-Disease
,	NN	O	O
are	NN	O	O
reported	NN	O	O
.	NN	O	O

Their	NN	O	O
parents	NN	O	O
originated	NN	O	O
from	NN	O	O
Calabria	NN	O	O
,	NN	O	O
from	NN	O	O
Sardinia	NN	O	O
and	NN	O	O
from	NN	O	O
Sicily	NN	O	O
.	NN	O	O

The	NN	O	O
abnormal	NN	O	O
enzymes	NN	O	O
are	NN	O	O
respectively	NN	O	O
designated	NN	O	O
as	NN	O	O
GdDcbrousse	NN	O	O
-	NN	O	O
like	NN	O	O
,	NN	O	O
GdGallura	NN	O	O
and	NN	O	O
GdAgrigento	NN	O	O
.	NN	O	O
.	NN	O	O

Cytogenetic	NN	O	O
investigations	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
.	NN	O	O

Chromosomal	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
on	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
five	NN	O	O
Israeli	NN	O	O
-	NN	O	O
Moroccan	NN	O	O
families	NN	O	O
with	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
24	NN	O	O
individuals	NN	O	O
,	NN	O	O
including	NN	O	O
seven	NN	O	O
propositi	NN	O	O
,	NN	O	O
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

Among	NN	O	O
the	NN	O	O
probands	NN	O	O
,	NN	O	O
significantly	NN	O	O
elevated	NN	O	O
rates	NN	O	O
of	NN	O	O
chromosome	NN	O	O
damage	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
both	NN	O	O
blood	NN	O	O
and	NN	O	O
skin	NN	O	O
.	NN	O	O

Skin	NN	O	O
fibroblasts	NN	O	O
of	NN	O	O
affected	NN	O	O
individuals	NN	O	O
showed	NN	O	O
several	NN	O	O
orders	NN	O	O
of	NN	O	O
magnitude	NN	O	O
more	NN	O	O
chromosome	NN	O	O
breakage	NN	O	O
than	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Increased	NN	O	O
rates	NN	O	O
of	NN	O	O
chromosome	NN	O	O
damage	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
fibroblasts	NN	O	O
of	NN	O	O
some	NN	O	O
phenotypically	NN	O	O
normal	NN	O	O
family	NN	O	O
members	NN	O	O
(	NN	O	O
obligate	NN	O	O
heterozygotes	NN	O	O
and	NN	O	O
sibs	NN	O	O
)	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

An	NN	O	O
apparent	NN	O	O
abnormal	NN	O	O
clone	NN	O	O
of	NN	O	O
cells	NN	O	O
,	NN	O	O
possessing	NN	O	O
a	NN	O	O
large	NN	O	O
acrocentric	NN	O	O
marker	NN	O	O
chromosome	NN	O	O
(	NN	O	O
14q	NN	O	O
+	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
varying	NN	O	O
proportions	NN	O	O
among	NN	O	O
cells	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
propositi	NN	O	O
(	NN	O	O
2	NN	O	O
-	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
;	NN	O	O
1	NN	O	O
-	NN	O	O
9	NN	O	O
%	NN	O	O
of	NN	O	O
fibroblasts	NN	O	O
)	NN	O	O
.	NN	O	O
.	NN	O	O

Prenatal	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Wolman	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
pregnancies	NN	O	O
at	NN	O	O
risk	NN	O	O
for	NN	O	O
Wolman	NN	O	B-Disease
disease	NN	O	I-Disease
were	NN	O	O
monitored	NN	O	O
by	NN	O	O
assay	NN	O	O
and	NN	O	O
electrophoresis	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
in	NN	O	O
cultured	NN	O	O
amniotic	NN	O	O
-	NN	O	O
fluid	NN	O	O
cells	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
patient	NN	O	O
1	NN	O	O
had	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
control	NN	O	O
levels	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
,	NN	O	O
using	NN	O	O
14C	NN	O	O
-	NN	O	O
triolein	NN	O	O
as	NN	O	O
substrate	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
when	NN	O	O
artificial	NN	O	O
substrates	NN	O	O
(	NN	O	O
esters	NN	O	O
of	NN	O	O
4	NN	O	O
-	NN	O	O
methylumbelliferone	NN	O	O
and	NN	O	O
p	NN	O	O
-	NN	O	O
nitrophenol	NN	O	O
)	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
acid	NN	O	O
lipase	NN	O	O
,	NN	O	O
these	NN	O	O
cells	NN	O	O
had	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
control	NN	O	O
levels	NN	O	O
.	NN	O	O

Electrophoresis	NN	O	O
of	NN	O	O
cell	NN	O	O
extracts	NN	O	O
revealed	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	O
form	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Wolman	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
fetal	NN	O	O
tissues	NN	O	O
following	NN	O	O
prostaglandin	NN	O	O
termination	NN	O	O
of	NN	O	O
this	NN	O	O
pregnancy	NN	O	O
confirmed	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

Assay	NN	O	O
of	NN	O	O
fetal	NN	O	O
-	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
with	NN	O	O
14C	NN	O	O
-	NN	O	O
triolein	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
with	NN	O	O
artificial	NN	O	O
substrates	NN	O	O
,	NN	O	O
showed	NN	O	O
marked	NN	O	O
deficiency	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
activity	NN	O	O
.	NN	O	O

Electrophoresis	NN	O	O
of	NN	O	O
fetal	NN	O	O
-	NN	O	O
tissue	NN	O	O
extracts	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	O
form	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
.	NN	O	O

Amniotic	NN	O	O
-	NN	O	O
fluid	NN	O	O
cells	NN	O	O
from	NN	O	O
patient	NN	O	O
2	NN	O	O
showed	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
with	NN	O	O
all	NN	O	O
substrates	NN	O	O
tested	NN	O	O
;	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
pattern	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
was	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Wolman	NN	O	B-Disease
disease	NN	O	I-Disease
be	NN	O	O
made	NN	O	O
using	NN	O	O
the	NN	O	O
radioassay	NN	O	O
of	NN	O	O
acid	NN	O	O
lipase	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
electrophoresis	NN	O	O
.	NN	O	O
.	NN	O	O

Malignant	NN	O	B-Disease
neoplasms	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
families	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
.	NN	O	O

Ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
(	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
syndrome	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
a	NN	O	O
greatly	NN	O	O
increased	NN	O	O
incidence	NN	O	O
of	NN	O	O
malignant	NN	O	B-Disease
neoplasms	NN	O	I-Disease
in	NN	O	O
homozygous	NN	O	O
affected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Heterozygotes	NN	O	O
for	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
are	NN	O	O
thought	NN	O	O
to	NN	O	O
comprise	NN	O	O
about	NN	O	O
1	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
and	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
to	NN	O	O
know	NN	O	O
whether	NN	O	O
this	NN	O	O
gene	NN	O	O
also	NN	O	O
predisposes	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
carrier	NN	O	O
to	NN	O	O
cancers	NN	O	B-Disease
.	NN	O	O

Heterozygous	NN	O	O
carriers	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
are	NN	O	O
common	NN	O	O
among	NN	O	O
the	NN	O	O
close	NN	O	O
relatives	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
,	NN	O	O
although	NN	O	O
individual	NN	O	O
carriers	NN	O	O
cannot	NN	O	O
be	NN	O	O
identified	NN	O	O
by	NN	O	O
any	NN	O	O
clinical	NN	O	O
criterion	NN	O	O
or	NN	O	O
laboratory	NN	O	O
test	NN	O	O
.	NN	O	O

For	NN	O	O
this	NN	O	O
reason	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
incidence	NN	O	O
of	NN	O	O
death	NN	O	O
from	NN	O	O
malignant	NN	O	B-Disease
neoplasms	NN	O	I-Disease
in	NN	O	O
2	NN	O	O
families	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
to	NN	O	O
that	NN	O	O
expected	NN	O	O
in	NN	O	O
a	NN	O	O
random	NN	O	O
sample	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
59	NN	O	O
deaths	NN	O	O
from	NN	O	O
malignant	NN	O	B-Disease
neoplasms	NN	O	I-Disease
in	NN	O	O
relatives	NN	O	O
dying	NN	O	O
before	NN	O	O
age	NN	O	O
75	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
42	NN	O	O
.	NN	O	O

6	NN	O	O
expected	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0	NN	O	O
.	NN	O	O
02	NN	O	O
)	NN	O	O
.	NN	O	O

For	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
heterozygotes	NN	O	O
younger	NN	O	O
than	NN	O	O
age	NN	O	O
45	NN	O	O
,	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
dying	NN	O	O
from	NN	O	O
a	NN	O	O
malignant	NN	O	B-Disease
neoplasm	NN	O	I-Disease
was	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
greater	NN	O	O
than	NN	O	O
5	NN	O	O
times	NN	O	O
the	NN	O	O
risk	NN	O	O
for	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O

A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
heterozygotes	NN	O	O
may	NN	O	O
comprise	NN	O	O
more	NN	O	O
than	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
all	NN	O	O
persons	NN	O	O
dying	NN	O	O
from	NN	O	O
a	NN	O	O
cancer	NN	O	B-Disease
before	NN	O	O
age	NN	O	O
45	NN	O	O
.	NN	O	O

The	NN	O	O
incidence	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
,	NN	O	I-Disease
gastric	NN	O	I-Disease
,	NN	O	I-Disease
and	NN	O	I-Disease
biliary	NN	O	I-Disease
system	NN	O	I-Disease
carcinomas	NN	O	I-Disease
and	NN	O	O
of	NN	O	O
leukemia	NN	O	B-Disease
and	NN	O	O
lymphoma	NN	O	B-Disease
was	NN	O	O
increased	NN	O	O
in	NN	O	O
these	NN	O	O
A	NN	O	B-Disease
-	NN	O	I-Disease
T	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

Other	NN	O	O
neoplasms	NN	O	B-Disease
that	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
heterozygotes	NN	O	O
include	NN	O	O
pancreatic	NN	O	B-Disease
,	NN	O	I-Disease
basal	NN	O	I-Disease
cell	NN	O	I-Disease
,	NN	O	I-Disease
colonic	NN	O	I-Disease
,	NN	O	I-Disease
breast	NN	O	I-Disease
,	NN	O	I-Disease
and	NN	O	I-Disease
cervical	NN	O	I-Disease
carcinomas	NN	O	I-Disease
.	NN	O	O

Striking	NN	O	O
prevalence	NN	O	O
of	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
in	NN	O	O
"	NN	O	O
healthy	NN	O	O
"	NN	O	O
w27	NN	O	O
positive	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O
.	NN	O	O

Ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
is	NN	O	O
diagnosed	NN	O	O
once	NN	O	O
or	NN	O	O
twice	NN	O	O
in	NN	O	O
each	NN	O	O
1000	NN	O	O
males	NN	O	O
and	NN	O	O
one	NN	O	O
tenth	NN	O	O
as	NN	O	O
frequently	NN	O	O
in	NN	O	O
females	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
true	NN	O	O
prevalence	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Indentification	NN	O	O
of	NN	O	O
genetic	NN	O	O
marker	NN	O	O
,	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
W27	NN	O	O
,	NN	O	O
for	NN	O	O
susceptible	NN	O	O
persons	NN	O	O
has	NN	O	O
provided	NN	O	O
a	NN	O	O
tool	NN	O	O
facilitating	NN	O	O
epidemiologic	NN	O	O
studies	NN	O	O
and	NN	O	O
allowing	NN	O	O
identification	NN	O	O
of	NN	O	O
"	NN	O	O
control	NN	O	O
"	NN	O	O
populations	NN	O	O
without	NN	O	O
the	NN	O	O
marker	NN	O	O
.	NN	O	O

Evaluation	NN	O	O
by	NN	O	O
postal	NN	O	O
questionnaires	NN	O	O
,	NN	O	O
and	NN	O	O
pelvic	NN	O	O
radiography	NN	O	O
of	NN	O	O
78	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
27W	NN	O	O
-	NN	O	O
positive	NN	O	O
blood	NN	O	O
donors	NN	O	O
selected	NN	O	O
from	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
apparently	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
revealed	NN	O	O
14	NN	O	O
who	NN	O	O
satisfied	NN	O	O
the	NN	O	O
criteria	NN	O	O
for	NN	O	O
definite	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
prevalence	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
sexes	NN	O	O
.	NN	O	O

One	NN	O	O
hundred	NN	O	O
and	NN	O	O
twenty	NN	O	O
-	NN	O	O
six	NN	O	O
W27	NN	O	O
-	NN	O	O
negative	NN	O	O
controls	NN	O	O
matched	NN	O	O
for	NN	O	O
race	NN	O	O
,	NN	O	O
sex	NN	O	O
,	NN	O	O
and	NN	O	O
age	NN	O	O
failed	NN	O	O
to	NN	O	O
yield	NN	O	O
a	NN	O	O
single	NN	O	O
case	NN	O	O
.	NN	O	O

For	NN	O	O
a	NN	O	O
person	NN	O	O
of	NN	O	O
either	NN	O	O
sex	NN	O	O
with	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
W27	NN	O	O
,	NN	O	O
there	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
about	NN	O	O
a	NN	O	O
20	NN	O	O
per	NN	O	O
cent	NN	O	O
chance	NN	O	O
that	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
will	NN	O	O
develop	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
prevalence	NN	O	O
of	NN	O	O
10	NN	O	O
to	NN	O	O
15	NN	O	O
per	NN	O	O
thousand	NN	O	O
.	NN	O	O

Hitherto	NN	O	O
accepted	NN	O	O
figures	NN	O	O
may	NN	O	O
underestimate	NN	O	O
the	NN	O	O
frequency	NN	O	O
by	NN	O	O
a	NN	O	O
factor	NN	O	O
of	NN	O	O
10	NN	O	O
to	NN	O	O
20	NN	O	O
.	NN	O	O
.	NN	O	O

Analbuminemia	NN	O	B-Disease
in	NN	O	O
an	NN	O	O
American	NN	O	O
Indian	NN	O	O
girl	NN	O	O
.	NN	O	O

Analbuminemia	NN	O	B-Disease
was	NN	O	O
fortuitously	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
nonedematous	NN	O	O
12	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
American	NN	O	O
Indian	NN	O	O
girl	NN	O	O
with	NN	O	O
atopic	NN	O	B-Disease
dermatitis	NN	O	I-Disease
,	NN	O	O
mild	NN	O	O
bronchial	NN	O	B-Disease
asthma	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
mild	NN	O	O
seizure	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hyperlipoproteinemia	NN	O	B-Disease
with	NN	O	O
a	NN	O	O
corneal	NN	O	B-Disease
arcus	NN	O	I-Disease
.	NN	O	O

Immunologic	NN	O	O
methods	NN	O	O
revealed	NN	O	O
trace	NN	O	O
amounts	NN	O	O
(	NN	O	O
17	NN	O	O
mg	NN	O	O
/	NN	O	O
100	NN	O	O
ml	NN	O	O
)	NN	O	O
of	NN	O	O
apparently	NN	O	O
normal	NN	O	O
serum	NN	O	O
albumin	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
parents	NN	O	O
were	NN	O	O
remotely	NN	O	O
related	NN	O	O
.	NN	O	O

The	NN	O	O
pedigree	NN	O	O
and	NN	O	O
clinical	NN	O	O
findings	NN	O	O
were	NN	O	O
compatible	NN	O	O
with	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
transmission	NN	O	O
of	NN	O	O
analbuminemia	NN	O	B-Disease
.	NN	O	O

Heterozygotes	NN	O	O
had	NN	O	O
subnormal	NN	O	O
levels	NN	O	O
of	NN	O	O
serum	NN	O	O
albumin	NN	O	O
.	NN	O	O

The	NN	O	O
Gc	NN	O	O
-	NN	O	O
locus	NN	O	O
is	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
structural	NN	O	O
albumin	NN	O	O
locus	NN	O	O
.	NN	O	O

Gc	NN	O	O
-	NN	O	O
protein	NN	O	O
levels	NN	O	O
were	NN	O	O
normal	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
together	NN	O	O
with	NN	O	O
normal	NN	O	O
chromosomal	NN	O	O
banding	NN	O	O
studies	NN	O	O
make	NN	O	O
it	NN	O	O
unlikely	NN	O	O
that	NN	O	O
a	NN	O	O
chromosomal	NN	O	O
deletion	NN	O	O
caused	NN	O	O
analbuminemia	NN	O	B-Disease
.	NN	O	O

Gc	NN	O	O
-	NN	O	O
types	NN	O	O
in	NN	O	O
the	NN	O	O
family	NN	O	O
were	NN	O	O
compatible	NN	O	O
with	NN	O	O
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
prove	NN	O	O
,	NN	O	O
linkage	NN	O	O
of	NN	O	O
analbuminemia	NN	O	B-Disease
to	NN	O	O
the	NN	O	O
Gc	NN	O	O
-	NN	O	O
locus	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
a	NN	O	O
"	NN	O	O
thalassemia	NN	O	B-Disease
"	NN	O	O
-	NN	O	O
like	NN	O	O
mutation	NN	O	O
for	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
KIT	NN	O	O
and	NN	O	O
PDGFRA	NN	O	O
genes	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
piebaldism	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
human	NN	O	O
piebaldism	NN	O	B-Disease
results	NN	O	O
from	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
KIT	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
the	NN	O	O
receptor	NN	O	O
for	NN	O	O
the	NN	O	O
mast	NN	O	O
/	NN	O	O
stem	NN	O	O
cell	NN	O	O
growth	NN	O	O
factor	NN	O	O
and	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
chromosome	NN	O	O
segment	NN	O	O
4q12	NN	O	O
.	NN	O	O

Using	NN	O	O
DNA	NN	O	O
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
piebaldism	NN	O	B-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
multiple	NN	O	B-Disease
congenital	NN	O	I-Disease
anomalies	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
a	NN	O	O
46	NN	O	O
,	NN	O	O
XY	NN	O	O
,	NN	O	O
del	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
(	NN	O	O
q12q21	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
karyotype	NN	O	O
,	NN	O	O
we	NN	O	O
carried	NN	O	O
out	NN	O	O
quantitative	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
KIT	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
adjacent	NN	O	O
PDGFRA	NN	O	O
(	NN	O	O
platelet	NN	O	O
-	NN	O	O
derived	NN	O	O
growth	NN	O	O
factor	NN	O	O
receptor	NN	O	O
alpha	NN	O	O
subunit	NN	O	O
)	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
hemizygous	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
KIT	NN	O	O
and	NN	O	O
PDGFRA	NN	O	O
genes	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
are	NN	O	O
included	NN	O	O
within	NN	O	O
the	NN	O	O
deleted	NN	O	O
region	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
KIT	NN	O	O
and	NN	O	O
PDGFRA	NN	O	O
genes	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
piebald	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O

A	NN	O	O
germ	NN	O	O
line	NN	O	O
mutation	NN	O	O
within	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
for	NN	O	O
the	NN	O	O
putative	NN	O	O
5	NN	O	O
-	NN	O	O
phosphoribosyl	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
pyrophosphate	NN	O	O
binding	NN	O	O
site	NN	O	O
of	NN	O	O
hypoxanthine	NN	O	O
-	NN	O	O
guanine	NN	O	O
phosphoribosyltransferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patient	NN	O	O
:	NN	O	O
missense	NN	O	O
mutations	NN	O	O
within	NN	O	O
a	NN	O	O
functionally	NN	O	O
important	NN	O	O
region	NN	O	O
probably	NN	O	O
cause	NN	O	O
disease	NN	O	O
.	NN	O	O

Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
complete	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
hypoxanthine	NN	O	I-Disease
guanine	NN	O	I-Disease
phosphoribosyltransferase	NN	O	I-Disease
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
germ	NN	O	O
line	NN	O	O
mutations	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
each	NN	O	O
mutant	NN	O	O
gene	NN	O	O
provides	NN	O	O
valuable	NN	O	O
information	NN	O	O
as	NN	O	O
to	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
that	NN	O	O
occurs	NN	O	O
spontaneously	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
HPRT	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
patient	NN	O	O
with	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
gene	NN	O	O
,	NN	O	O
designated	NN	O	O
HPRT	NN	O	O
Tokyo	NN	O	O
,	NN	O	O
had	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
change	NN	O	O
from	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
,	NN	O	O
as	NN	O	O
identified	NN	O	O
by	NN	O	O
sequencing	NN	O	O
cDNA	NN	O	O
amplified	NN	O	O
by	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

Allele	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
hybridization	NN	O	O
analysis	NN	O	O
using	NN	O	O
amplified	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
mutant	NN	O	O
gene	NN	O	O
was	NN	O	O
transmitted	NN	O	O
from	NN	O	O
the	NN	O	O
maternal	NN	O	O
germ	NN	O	O
line	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
would	NN	O	O
lead	NN	O	O
to	NN	O	O
an	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
of	NN	O	O
Asp	NN	O	O
for	NN	O	O
Gly	NN	O	O
at	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
position	NN	O	O
140	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
putative	NN	O	O
5	NN	O	O
-	NN	O	O
phosphoribosyl	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
pyrophosphate	NN	O	O
(	NN	O	O
PRPP	NN	O	O
)	NN	O	O
binding	NN	O	O
region	NN	O	O
.	NN	O	O

Missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
human	NN	O	O
HPRT	NN	O	B-Disease
deficient	NN	O	I-Disease
patients	NN	O	O
thus	NN	O	O
far	NN	O	O
reported	NN	O	O
tend	NN	O	O
to	NN	O	O
accumulate	NN	O	O
in	NN	O	O
this	NN	O	O
functionally	NN	O	O
active	NN	O	O
region	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
data	NN	O	O
suggested	NN	O	O
that	NN	O	O
both	NN	O	O
missense	NN	O	O
and	NN	O	O
synonymous	NN	O	O
mutations	NN	O	O
can	NN	O	O
occur	NN	O	O
at	NN	O	O
any	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
germ	NN	O	O
line	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
,	NN	O	O
but	NN	O	O
that	NN	O	O
a	NN	O	O
limited	NN	O	O
percentage	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
missense	NN	O	O
mutations	NN	O	O
cause	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
probability	NN	O	O
that	NN	O	O
a	NN	O	O
mutation	NN	O	O
will	NN	O	O
cause	NN	O	O
disease	NN	O	O
tends	NN	O	O
to	NN	O	O
be	NN	O	O
higher	NN	O	O
when	NN	O	O
the	NN	O	O
missense	NN	O	O
mutation	NN	O	O
is	NN	O	O
within	NN	O	O
a	NN	O	O
functionally	NN	O	O
important	NN	O	O
sequence	NN	O	O
.	NN	O	O
.	NN	O	O

Characterisation	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
rare	NN	O	O
fragile	NN	O	O
site	NN	O	O
easily	NN	O	O
confused	NN	O	O
with	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
new	NN	O	O
fragile	NN	O	O
site	NN	O	O
(	NN	O	O
FRAXE	NN	O	O
)	NN	O	O
in	NN	O	O
Xq28	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
typical	NN	O	O
folate	NN	O	O
sensitive	NN	O	O
fragile	NN	O	O
site	NN	O	O
.	NN	O	O

The	NN	O	O
fragile	NN	O	O
site	NN	O	O
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
give	NN	O	O
abnormal	NN	O	O
results	NN	O	O
when	NN	O	O
subjected	NN	O	O
to	NN	O	O
Southern	NN	O	O
analysis	NN	O	O
with	NN	O	O
probe	NN	O	O
pfxa3	NN	O	O
which	NN	O	O
detects	NN	O	O
the	NN	O	O
unstable	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
characteristic	NN	O	O
of	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
mapping	NN	O	O
locates	NN	O	O
the	NN	O	O
fragile	NN	O	O
site	NN	O	O
between	NN	O	O
150	NN	O	O
kb	NN	O	O
and	NN	O	O
600	NN	O	O
kb	NN	O	O
distal	NN	O	O
to	NN	O	O
FRAXA	NN	O	O
.	NN	O	O

The	NN	O	O
distinction	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
fragile	NN	O	O
sites	NN	O	O
is	NN	O	O
important	NN	O	O
clinically	NN	O	O
since	NN	O	O
cytogenetic	NN	O	O
detection	NN	O	O
of	NN	O	O
FRAXE	NN	O	O
,	NN	O	O
without	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
,	NN	O	O
could	NN	O	O
result	NN	O	O
in	NN	O	O
misdiagnosis	NN	O	O
of	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

The	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
maps	NN	O	O
to	NN	O	O
a	NN	O	O
150	NN	O	O
kb	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
Xp11	NN	O	O
.	NN	O	O
3	NN	O	O
.	NN	O	O

Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
human	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	O
unknown	NN	O	O
etiology	NN	O	O
characterized	NN	O	O
by	NN	O	O
congenital	NN	O	B-Disease
blindness	NN	O	I-Disease
,	NN	O	O
sensory	NN	O	B-Disease
neural	NN	O	I-Disease
deafness	NN	O	I-Disease
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
disease	NN	O	O
gene	NN	O	O
was	NN	O	O
previously	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
DXS7	NN	O	O
(	NN	O	O
L1	NN	O	O
.	NN	O	O
28	NN	O	O
)	NN	O	O
locus	NN	O	O
and	NN	O	O
the	NN	O	O
MAO	NN	O	O
genes	NN	O	O
in	NN	O	O
band	NN	O	O
Xp11	NN	O	O
.	NN	O	O

3	NN	O	O
3	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
fine	NN	O	O
physical	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
obligate	NN	O	O
region	NN	O	O
containing	NN	O	O
the	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
NDP	NN	O	O
)	NN	O	O
defined	NN	O	O
by	NN	O	O
a	NN	O	O
recombination	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
smallest	NN	O	O
submicroscopic	NN	O	O
chromosomal	NN	O	O
deletion	NN	O	O
associated	NN	O	O
with	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
identified	NN	O	O
to	NN	O	O
date	NN	O	O
.	NN	O	O

Analysis	NN	O	O
,	NN	O	O
using	NN	O	O
in	NN	O	O
addition	NN	O	O
two	NN	O	O
overlapping	NN	O	O
YAC	NN	O	O
clones	NN	O	O
from	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
allowed	NN	O	O
orientation	NN	O	O
of	NN	O	O
the	NN	O	O
MAOA	NN	O	O
and	NN	O	O
MAOB	NN	O	O
genes	NN	O	O
in	NN	O	O
a	NN	O	O
5	NN	O	O
-	NN	O	O
3	NN	O	O
-	NN	O	O
3	NN	O	O
-	NN	O	O
5	NN	O	O
configuration	NN	O	O
.	NN	O	O

A	NN	O	O
recombination	NN	O	O
event	NN	O	O
between	NN	O	O
a	NN	O	O
(	NN	O	O
GT	NN	O	O
)	NN	O	O
n	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
intron	NN	O	O
2	NN	O	O
of	NN	O	O
the	NN	O	O
MAOB	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
NDP	NN	O	O
locus	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
previously	NN	O	O
reported	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
recombination	NN	O	O
between	NN	O	O
DXS7	NN	O	O
and	NN	O	O
NDP	NN	O	O
,	NN	O	O
delineates	NN	O	O
a	NN	O	O
flanking	NN	O	O
marker	NN	O	O
telomeric	NN	O	O
to	NN	O	O
this	NN	O	O
disease	NN	O	O
gene	NN	O	O
.	NN	O	O

An	NN	O	O
anonymous	NN	O	O
DNA	NN	O	O
probe	NN	O	O
,	NN	O	O
dc12	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
YACs	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
submicroscopic	NN	O	O
deletion	NN	O	O
which	NN	O	O
includes	NN	O	O
MAOA	NN	O	O
and	NN	O	O
MAOB	NN	O	O
but	NN	O	O
not	NN	O	O
L1	NN	O	O
.	NN	O	O

28	NN	O	O
,	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
flanking	NN	O	O
marker	NN	O	O
centromeric	NN	O	O
to	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
.	NN	O	O

An	NN	O	O
Alu	NN	O	O
-	NN	O	O
PCR	NN	O	O
fragment	NN	O	O
from	NN	O	O
the	NN	O	O
right	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
MAO	NN	O	O
YAC	NN	O	O
(	NN	O	O
YMAO	NN	O	O
.	NN	O	O
AluR	NN	O	O
)	NN	O	O
is	NN	O	O
not	NN	O	O
deleted	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
and	NN	O	O
also	NN	O	O
delineates	NN	O	O
the	NN	O	O
centromeric	NN	O	O
extent	NN	O	O
of	NN	O	O
the	NN	O	O
obligate	NN	O	O
disease	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
order	NN	O	O
of	NN	O	O
these	NN	O	O
loci	NN	O	O
is	NN	O	O
telomere	NN	O	O
.	NN	O	O

DXS7	NN	O	O
-	NN	O	O
MAOA	NN	O	O
-	NN	O	O
MAOB	NN	O	O
-	NN	O	O
NDP	NN	O	O
-	NN	O	O
dc12	NN	O	O
-	NN	O	O
YMAO	NN	O	O
DXS7	NN	O	O
-	NN	O	O
MAOA	NN	O	O
-	NN	O	O
MAOB	NN	O	O
-	NN	O	O
NDP	NN	O	O
-	NN	O	O
dc12	NN	O	O
-	NN	O	O
YMAO	NN	O	O
.	NN	O	O

AluR	NN	O	O
.	NN	O	O
centromere	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
data	NN	O	O
define	NN	O	O
the	NN	O	O
obligate	NN	O	O
region	NN	O	O
containing	NN	O	O
the	NN	O	O
NDP	NN	O	O
gene	NN	O	O
to	NN	O	O
a	NN	O	O
chromosomal	NN	O	O
segment	NN	O	O
less	NN	O	O
than	NN	O	O
150	NN	O	O
kb	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
phenylketonuria	NN	O	B-Disease
and	NN	O	O
related	NN	O	O
hyperphenylalaninemias	NN	O	B-Disease
:	NN	O	O
mutations	NN	O	O
and	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
producing	NN	O	O
phenylketonuria	NN	O	B-Disease
or	NN	O	O
hyperphenylalaninemia	NN	O	B-Disease
have	NN	O	O
now	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
many	NN	O	O
patients	NN	O	O
from	NN	O	O
various	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
all	NN	O	O
exhibit	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
association	NN	O	O
with	NN	O	O
specific	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
-	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
haplotypes	NN	O	O
at	NN	O	O
the	NN	O	O
PAH	NN	O	O
locus	NN	O	O
.	NN	O	O

About	NN	O	O
50	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
are	NN	O	O
single	NN	O	O
-	NN	O	O
base	NN	O	O
substitutions	NN	O	O
,	NN	O	O
including	NN	O	O
six	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
and	NN	O	O
eight	NN	O	O
splicing	NN	O	O
mutations	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
remainder	NN	O	O
being	NN	O	O
missense	NN	O	O
mutations	NN	O	O
.	NN	O	O

One	NN	O	O
splicing	NN	O	O
mutation	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
3	NN	O	O
amino	NN	O	O
acid	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
insertion	NN	O	O
.	NN	O	O

Two	NN	O	O
or	NN	O	O
3	NN	O	O
large	NN	O	O
deletions	NN	O	O
,	NN	O	O
2	NN	O	O
single	NN	O	O
codon	NN	O	O
deletions	NN	O	O
,	NN	O	O
and	NN	O	O
2	NN	O	O
single	NN	O	O
base	NN	O	O
deletions	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
.	NN	O	O

Twelve	NN	O	O
of	NN	O	O
the	NN	O	O
missense	NN	O	O
mutations	NN	O	O
apparently	NN	O	O
result	NN	O	O
from	NN	O	O
the	NN	O	O
methylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
deamination	NN	O	O
of	NN	O	O
highly	NN	O	O
mutagenic	NN	O	O
CpG	NN	O	O
dinucleotides	NN	O	O
.	NN	O	O

Recurrent	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
at	NN	O	O
several	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
,	NN	O	O
producing	NN	O	O
associations	NN	O	O
with	NN	O	O
different	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
different	NN	O	O
populations	NN	O	O
.	NN	O	O

About	NN	O	O
half	NN	O	O
of	NN	O	O
all	NN	O	O
missense	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
examined	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
expression	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
between	NN	O	O
residual	NN	O	O
PAH	NN	O	O
activity	NN	O	O
and	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Since	NN	O	O
continuing	NN	O	O
advances	NN	O	O
in	NN	O	O
molecular	NN	O	O
methodologies	NN	O	O
have	NN	O	O
dramatically	NN	O	O
accelerated	NN	O	O
the	NN	O	O
rate	NN	O	O
in	NN	O	O
which	NN	O	O
new	NN	O	O
mutations	NN	O	O
are	NN	O	O
being	NN	O	O
identified	NN	O	O
and	NN	O	O
characterized	NN	O	O
,	NN	O	O
this	NN	O	O
register	NN	O	O
of	NN	O	O
mutations	NN	O	O
will	NN	O	O
be	NN	O	O
updated	NN	O	O
periodically	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
glycine250	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
aspartate	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
of	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
causes	NN	O	O
juvenile	NN	O	O
-	NN	O	O
onset	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
Lebanese	NN	O	O
-	NN	O	O
Canadian	NN	O	O
family	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
causing	NN	O	O
juvenile	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
two	NN	O	O
sibs	NN	O	O
of	NN	O	O
Lebanese	NN	O	O
-	NN	O	O
Maronite	NN	O	O
origin	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

An	NN	O	O
mRNA	NN	O	O
-	NN	O	O
containing	NN	O	O
extract	NN	O	O
of	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
obtained	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
probands	NN	O	O
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
template	NN	O	O
to	NN	O	O
amplify	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
(	NN	O	O
Hex	NN	O	O
A	NN	O	O
)	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
amplified	NN	O	O
cDNA	NN	O	O
fragments	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
alteration	NN	O	O
,	NN	O	O
guanine	NN	O	O
to	NN	O	O
adenine	NN	O	O
at	NN	O	O
nt	NN	O	O
749	NN	O	O
creating	NN	O	O
a	NN	O	O
G250D	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
introduces	NN	O	O
a	NN	O	O
new	NN	O	O
recognition	NN	O	O
site	NN	O	O
for	NN	O	O
the	NN	O	O
restriction	NN	O	O
enzyme	NN	O	O
Eco	NN	O	O
RV	NN	O	O
,	NN	O	O
permitting	NN	O	O
identification	NN	O	O
of	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
this	NN	O	O
allele	NN	O	O
following	NN	O	O
PCR	NN	O	O
amplification	NN	O	O
and	NN	O	O
Eco	NN	O	O
RV	NN	O	O
digestion	NN	O	O
of	NN	O	O
exon	NN	O	O
7	NN	O	O
sequences	NN	O	O
from	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
templates	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
test	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
substitution	NN	O	O
,	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
mutagenized	NN	O	O
cDNA	NN	O	O
construct	NN	O	O
was	NN	O	O
introduced	NN	O	O
into	NN	O	O
a	NN	O	O
mammalian	NN	O	O
expression	NN	O	O
vector	NN	O	O
and	NN	O	O
transfected	NN	O	O
into	NN	O	O
monkey	NN	O	O
Cos	NN	O	O
-	NN	O	O
1	NN	O	O
cells	NN	O	O
separately	NN	O	O
or	NN	O	O
along	NN	O	O
with	NN	O	O
a	NN	O	O
beta	NN	O	O
-	NN	O	O
cDNA	NN	O	O
expression	NN	O	O
vector	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
mutant	NN	O	O
alpha	NN	O	O
-	NN	O	O
cDNA	NN	O	O
was	NN	O	O
the	NN	O	O
only	NN	O	O
gene	NN	O	O
introduced	NN	O	O
into	NN	O	O
COS	NN	O	O
cells	NN	O	O
no	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
above	NN	O	O
endogenous	NN	O	O
COS	NN	O	O
cell	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
normal	NN	O	O
alpha	NN	O	O
-	NN	O	O
cDNA	NN	O	O
and	NN	O	O
beta	NN	O	O
-	NN	O	O
cDNA	NN	O	O
followed	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
of	NN	O	O
human	NN	O	O
Hex	NN	O	O
A	NN	O	O
resulted	NN	O	O
in	NN	O	O
20	NN	O	O
-	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
ratio	NN	O	O
between	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
(	NN	O	O
mock	NN	O	O
transfection	NN	O	O
)	NN	O	O
control	NN	O	O
values	NN	O	O
.	NN	O	O

This	NN	O	O
allowed	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
some	NN	O	O
residual	NN	O	O
activity	NN	O	O
(	NN	O	O
12	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
positive	NN	O	O
control	NN	O	O
)	NN	O	O
when	NN	O	O
the	NN	O	O
mutant	NN	O	O
alpha	NN	O	O
-	NN	O	O
cDNA	NN	O	O
replaced	NN	O	O
its	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
counterpart	NN	O	O
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
protein	NN	O	O
environment	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
is	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
mutation	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
Gly269	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Ser	NN	O	O
substitution	NN	O	O
in	NN	O	O
exon	NN	O	O
7	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O
.	NN	O	O

Novel	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
mutations	NN	O	O
from	NN	O	O
China	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
three	NN	O	O
HEXA	NN	O	O
mutations	NN	O	O
associated	NN	O	O
with	NN	O	O
infantile	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
three	NN	O	O
unrelated	NN	O	O
nonconsanguineous	NN	O	O
Chinese	NN	O	O
families	NN	O	O
.	NN	O	O

Novel	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
families	NN	O	O
.	NN	O	O

The	NN	O	O
third	NN	O	O
is	NN	O	O
a	NN	O	O
previously	NN	O	O
reported	NN	O	O
mutation	NN	O	O
(	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
nt	NN	O	O
1444	NN	O	O
)	NN	O	O
(	NN	O	O
Nakano	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1988	NN	O	O
)	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
PCR	NN	O	O
products	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
insertion	NN	O	O
of	NN	O	O
an	NN	O	O
A	NN	O	O
after	NN	O	O
nt	NN	O	O
547	NN	O	O
in	NN	O	O
family	NN	O	O
1	NN	O	O
.	NN	O	O

This	NN	O	O
change	NN	O	O
generates	NN	O	O
an	NN	O	O
early	NN	O	O
termination	NN	O	O
codon	NN	O	O
6	NN	O	O
bp	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
insertion	NN	O	O
site	NN	O	O
.	NN	O	O

Allele	NN	O	O
-	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
hybridization	NN	O	O
confirmed	NN	O	O
homozygosity	NN	O	O
in	NN	O	O
the	NN	O	O
proband	NN	O	O
.	NN	O	O

Single	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
amplified	NN	O	O
exon	NN	O	O
13	NN	O	O
revealed	NN	O	O
a	NN	O	O
T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
transition	NN	O	O
at	NN	O	O
nt	NN	O	O
1453	NN	O	O
with	NN	O	O
the	NN	O	O
corresponding	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
W485R	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
family	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
creates	NN	O	O
an	NN	O	O
Fnu4HI	NN	O	O
restriction	NN	O	O
site	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
is	NN	O	O
homozygous	NN	O	O
for	NN	O	O
this	NN	O	O
allele	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
site	NN	O	O
-	NN	O	O
specific	NN	O	O
mutagenized	NN	O	O
alpha	NN	O	O
cDNA	NN	O	O
carrying	NN	O	O
the	NN	O	O
T	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
transition	NN	O	O
at	NN	O	O
nt	NN	O	O
1453	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
COS	NN	O	O
1	NN	O	O
cells	NN	O	O
hexosaminidase	NN	O	O
S	NN	O	O
activity	NN	O	O
was	NN	O	O
not	NN	O	O
detectable	NN	O	O
above	NN	O	O
background	NN	O	O
.	NN	O	O

A	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
nt	NN	O	O
1444	NN	O	O
(	NN	O	O
exon	NN	O	O
13	NN	O	O
)	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
E482K	NN	O	O
substitution	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	O
family	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
at	NN	O	O
a	NN	O	O
CpG	NN	O	O
dinucleotide	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
an	NN	O	O
Italian	NN	O	O
TSD	NN	O	B-Disease
proband	NN	O	O
and	NN	O	O
causes	NN	O	O
defective	NN	O	O
intracellular	NN	O	O
transport	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
from	NN	O	O
the	NN	O	O
rough	NN	O	O
endoplasmic	NN	O	O
reticulum	NN	O	O
to	NN	O	O
the	NN	O	O
Golgi	NN	O	O
apparatus	NN	O	O
.	NN	O	O

Two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
causing	NN	O	O
mild	NN	O	O
hyperphenylalaninemia	NN	O	B-Disease
associated	NN	O	O
with	NN	O	O
DNA	NN	O	O
haplotype	NN	O	O
12	NN	O	O
.	NN	O	O

The	NN	O	O
genetic	NN	O	B-Disease
defects	NN	O	I-Disease
responsible	NN	O	O
for	NN	O	O
most	NN	O	O
phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
hyperphenylalaninemia	NN	O	B-Disease
(	NN	O	O
HPA	NN	O	B-Disease
)	NN	O	O
cases	NN	O	O
are	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Approximately	NN	O	O
50	NN	O	O
-	NN	O	O
60	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
Caucasians	NN	O	O
and	NN	O	O
are	NN	O	O
reflected	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
clinical	NN	O	O
severities	NN	O	O
.	NN	O	O

Most	NN	O	O
mutations	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
specific	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
as	NN	O	O
defined	NN	O	O
by	NN	O	O
eight	NN	O	O
polymorphic	NN	O	O
restriction	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mild	NN	O	O
mutation	NN	O	O
linked	NN	O	O
to	NN	O	O
haplotype	NN	O	O
12	NN	O	O
in	NN	O	O
the	NN	O	O
Swedish	NN	O	O
PKU	NN	O	B-Disease
/	NN	O	O
HPA	NN	O	B-Disease
population	NN	O	O
,	NN	O	O
since	NN	O	O
7	NN	O	O
of	NN	O	O
8	NN	O	O
patients	NN	O	O
carrying	NN	O	O
haplotype	NN	O	O
12	NN	O	O
had	NN	O	O
mild	NN	O	O
HPA	NN	O	B-Disease
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
G	NN	O	O
transversion	NN	O	O
at	NN	O	O
the	NN	O	O
second	NN	O	O
base	NN	O	O
of	NN	O	O
codon	NN	O	O
322	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
substitution	NN	O	O
of	NN	O	O
glycine	NN	O	O
for	NN	O	O
alanine	NN	O	O
,	NN	O	O
in	NN	O	O
four	NN	O	O
mutant	NN	O	O
haplotype	NN	O	O
12	NN	O	O
genes	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
second	NN	O	O
base	NN	O	O
of	NN	O	O
codon	NN	O	O
408	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
substitution	NN	O	O
of	NN	O	O
glutamine	NN	O	O
for	NN	O	O
arginine	NN	O	O
,	NN	O	O
in	NN	O	O
another	NN	O	O
three	NN	O	O
mutant	NN	O	O
haplotype	NN	O	O
12	NN	O	O
genes	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
segregated	NN	O	O
with	NN	O	O
mutant	NN	O	O
haplotype	NN	O	O
12	NN	O	O
alleles	NN	O	O
in	NN	O	O
nuclear	NN	O	O
families	NN	O	O
but	NN	O	O
were	NN	O	O
not	NN	O	O
present	NN	O	O
on	NN	O	O
normal	NN	O	O
or	NN	O	O
other	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
were	NN	O	O
tested	NN	O	O
in	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
expression	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
enzyme	NN	O	O
activities	NN	O	O
of	NN	O	O
different	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
enzymes	NN	O	O
reflect	NN	O	O
the	NN	O	O
relative	NN	O	O
severities	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
,	NN	O	O
although	NN	O	O
these	NN	O	O
in	NN	O	O
vitro	NN	O	O
activities	NN	O	O
cannot	NN	O	O
be	NN	O	O
translated	NN	O	O
directly	NN	O	O
into	NN	O	O
in	NN	O	O
vivo	NN	O	O
hepatic	NN	O	O
activities	NN	O	O
.	NN	O	O

The	NN	O	O
A322G	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
had	NN	O	O
about	NN	O	O
75	NN	O	O
%	NN	O	O
and	NN	O	O
the	NN	O	O
R408Q	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
about	NN	O	O
55	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
PAH	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
in	NN	O	O
vitro	NN	O	O
activities	NN	O	O
are	NN	O	O
the	NN	O	O
highest	NN	O	O
reported	NN	O	O
for	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
enzymes	NN	O	O
produced	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
expression	NN	O	O
system	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
correlations	NN	O	O
for	NN	O	O
I65T	NN	O	O
and	NN	O	O
M1V	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
locus	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
locus	NN	O	O
are	NN	O	O
the	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
hyperphenylalaninemia	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
described	NN	O	O
four	NN	O	O
mutations	NN	O	O
(	NN	O	O
M1V	NN	O	O
,	NN	O	O
IVS12nt1	NN	O	O
,	NN	O	O
R408W	NN	O	O
,	NN	O	O
and	NN	O	O
S349P	NN	O	O
)	NN	O	O
at	NN	O	O
the	NN	O	O
PAH	NN	O	O
locus	NN	O	O
in	NN	O	O
French	NN	O	O
Canadians	NN	O	O
with	NN	O	O
ancestry	NN	O	O
in	NN	O	O
eastern	NN	O	O
Quebec	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
identification	NN	O	O
of	NN	O	O
another	NN	O	O
mutation	NN	O	O
,	NN	O	O
on	NN	O	O
a	NN	O	O
haplotype	NN	O	O
9	NN	O	O
chromosome	NN	O	O
,	NN	O	O
which	NN	O	O
converts	NN	O	O
codon	NN	O	O
65	NN	O	O
from	NN	O	O
isoleucine	NN	O	O
(	NN	O	O
ATT	NN	O	O
)	NN	O	O
to	NN	O	O
threonine	NN	O	O
(	NN	O	O
ACT	NN	O	O
)	NN	O	O
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
expression	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
I65T	NN	O	O
mutation	NN	O	O
in	NN	O	O
COS	NN	O	O
cells	NN	O	O
demonstrating	NN	O	O
75	NN	O	O
%	NN	O	O
loss	NN	O	O
of	NN	O	O
both	NN	O	O
immunoreactive	NN	O	O
protein	NN	O	O
and	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
expression	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
prevalent	NN	O	O
PKU	NN	O	B-Disease
allele	NN	O	O
(	NN	O	O
M1V	NN	O	O
)	NN	O	O
in	NN	O	O
eastern	NN	O	O
Quebec	NN	O	O
,	NN	O	O
showing	NN	O	O
nondetectable	NN	O	O
levels	NN	O	O
of	NN	O	O
PAH	NN	O	O
protein	NN	O	O
and	NN	O	O
activity	NN	O	O
,	NN	O	O
a	NN	O	O
finding	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
translation	NN	O	O
initiation	NN	O	O
codon	NN	O	O
.	NN	O	O

Homozygosity	NN	O	O
for	NN	O	O
M1V	NN	O	O
and	NN	O	O
codominant	NN	O	O
inheritance	NN	O	O
of	NN	O	O
I65T	NN	O	O
/	NN	O	O
R408W	NN	O	O
were	NN	O	O
both	NN	O	O
associated	NN	O	O
with	NN	O	O
classical	NN	O	B-Disease
phenylketonuria	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
hexosaminidase	NN	O	B-Disease
A	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
pseudodeficiency	NN	O	O
in	NN	O	O
the	NN	O	O
Berks	NN	O	O
County	NN	O	O
Pennsylvania	NN	O	O
Dutch	NN	O	O
.	NN	O	O

Following	NN	O	O
the	NN	O	O
birth	NN	O	O
of	NN	O	O
two	NN	O	O
infants	NN	O	O
with	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
,	NN	O	O
Pennsylvania	NN	O	O
Dutch	NN	O	O
kindred	NN	O	O
was	NN	O	O
screened	NN	O	O
for	NN	O	O
TSD	NN	O	B-Disease
carriers	NN	O	O
using	NN	O	O
the	NN	O	O
biochemical	NN	O	O
assay	NN	O	O
.	NN	O	O

A	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
individuals	NN	O	O
who	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
TSD	NN	O	B-Disease
heterozygotes	NN	O	O
was	NN	O	O
detected	NN	O	O
(	NN	O	O
Kelly	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1975	NN	O	O
)	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
evidence	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
increased	NN	O	O
carrier	NN	O	O
frequency	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
altered	NN	O	O
alleles	NN	O	O
for	NN	O	O
the	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
two	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
in	NN	O	O
this	NN	O	O
Pennsylvania	NN	O	O
Dutch	NN	O	O
kindred	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

One	NN	O	O
allele	NN	O	O
,	NN	O	O
reported	NN	O	O
originally	NN	O	O
in	NN	O	O
a	NN	O	O
French	NN	O	O
TSD	NN	O	B-Disease
patient	NN	O	O
(	NN	O	O
Akli	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
GT	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
AT	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
9	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
,	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
739	NN	O	O
(	NN	O	O
Arg247Trp	NN	O	O
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
by	NN	O	O
Triggs	NN	O	O
-	NN	O	O
Raine	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O

(	NN	O	O
1992	NN	O	O
)	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
clinically	NN	O	O
benign	NN	O	O
"	NN	O	O
pseudodeficient	NN	O	O
"	NN	O	O
allele	NN	O	O
associated	NN	O	O
with	NN	O	O
reduced	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
against	NN	O	O
artificial	NN	O	O
substrate	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
polymorphism	NN	O	O
[	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
(	NN	O	O
759	NN	O	O
)	NN	O	O
]	NN	O	O
,	NN	O	O
which	NN	O	O
leaves	NN	O	O
valine	NN	O	O
at	NN	O	O
codon	NN	O	O
253	NN	O	O
unchanged	NN	O	O
,	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

A	NN	O	O
mutation	NN	O	O
common	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
:	NN	O	O
frequency	NN	O	O
and	NN	O	O
RNA	NN	O	O
studies	NN	O	O
.	NN	O	O

Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
resulting	NN	O	O
from	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
enzyme	NN	O	O
,	NN	O	O
beta	NN	O	O
-	NN	O	O
N	NN	O	O
-	NN	O	O
acetylhexosaminidase	NN	O	O
A	NN	O	O
(	NN	O	O
Hex	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
discovered	NN	O	O
that	NN	O	O
a	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
mutation	NN	O	O
,	NN	O	O
IVS	NN	O	O
-	NN	O	O
9	NN	O	O
+	NN	O	O
1	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
first	NN	O	O
detected	NN	O	O
by	NN	O	O
Akli	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O

(	NN	O	O
Genomics	NN	O	O
11	NN	O	O
124	NN	O	O
-	NN	O	O
134	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
disease	NN	O	O
allele	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Caucasians	NN	O	O
(	NN	O	O
10	NN	O	O
/	NN	O	O
58	NN	O	O
alleles	NN	O	O
examined	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
PCR	NN	O	O
-	NN	O	O
based	NN	O	O
diagnostic	NN	O	O
test	NN	O	O
,	NN	O	O
which	NN	O	O
detects	NN	O	O
an	NN	O	O
NlaIII	NN	O	O
site	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
mutation	NN	O	O
,	NN	O	O
revealed	NN	O	O
a	NN	O	O
frequency	NN	O	O
among	NN	O	O
enzyme	NN	O	O
-	NN	O	O
defined	NN	O	O
carriers	NN	O	O
of	NN	O	O
9	NN	O	O
/	NN	O	O
64	NN	O	O
(	NN	O	O
14	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
those	NN	O	O
carrying	NN	O	O
the	NN	O	O
allele	NN	O	O
trace	NN	O	O
their	NN	O	O
origins	NN	O	O
to	NN	O	O
the	NN	O	O
United	NN	O	O
Kingdom	NN	O	O
,	NN	O	O
Ireland	NN	O	O
,	NN	O	O
or	NN	O	O
Western	NN	O	O
Europe	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
not	NN	O	O
identified	NN	O	O
among	NN	O	O
12	NN	O	O
Black	NN	O	O
American	NN	O	O
TSD	NN	O	B-Disease
alleles	NN	O	O
or	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
18	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
,	NN	O	O
enzyme	NN	O	O
-	NN	O	O
defined	NN	O	O
carriers	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
carry	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
common	NN	O	O
to	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

No	NN	O	O
normally	NN	O	O
spliced	NN	O	O
RNA	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
PCR	NN	O	O
products	NN	O	O
generated	NN	O	O
from	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
of	NN	O	O
RNA	NN	O	O
carrying	NN	O	O
the	NN	O	O
IVS	NN	O	O
-	NN	O	O
9	NN	O	O
mutation	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
the	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
mRNA	NN	O	O
from	NN	O	O
this	NN	O	O
allele	NN	O	O
were	NN	O	O
comprised	NN	O	O
of	NN	O	O
aberrant	NN	O	O
species	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
either	NN	O	O
of	NN	O	O
two	NN	O	O
cryptic	NN	O	O
donor	NN	O	O
sites	NN	O	O
,	NN	O	O
one	NN	O	O
truncating	NN	O	O
exon	NN	O	O
9	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
within	NN	O	O
IVS	NN	O	O
-	NN	O	O
9	NN	O	O
,	NN	O	O
spliced	NN	O	O
to	NN	O	O
exon	NN	O	O
10	NN	O	O
.	NN	O	O

Numerous	NN	O	O
additional	NN	O	O
splice	NN	O	O
products	NN	O	O
were	NN	O	O
detected	NN	O	O
,	NN	O	O
most	NN	O	O
involving	NN	O	O
skipping	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
surrounding	NN	O	O
exons	NN	O	O
.	NN	O	O

Together	NN	O	O
with	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
allele	NN	O	O
responsible	NN	O	O
for	NN	O	O
Hex	NN	O	O
A	NN	O	O
pseudodeficiency	NN	O	O
(	NN	O	O
Triggs	NN	O	O
-	NN	O	O
Raine	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
Am	NN	O	O
J	NN	O	O
Hum	NN	O	O
Genet	NN	O	O
,	NN	O	O
1992	NN	O	O
)	NN	O	O
,	NN	O	O
these	NN	O	O
two	NN	O	O
alleles	NN	O	O
accounted	NN	O	O
for	NN	O	O
almost	NN	O	O
50	NN	O	O
%	NN	O	O
(	NN	O	O
29	NN	O	O
/	NN	O	O
64	NN	O	O
)	NN	O	O
of	NN	O	O
TSD	NN	O	B-Disease
or	NN	O	O
carrier	NN	O	O
alleles	NN	O	O
ascertained	NN	O	O
by	NN	O	O
enzyme	NN	O	O
screening	NN	O	O
tests	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Caucasians	NN	O	O
.	NN	O	O
.	NN	O	O

Aberrant	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
a	NN	O	O
significant	NN	O	O
cause	NN	O	O
of	NN	O	O
choroideremia	NN	O	B-Disease
.	NN	O	O

Choroideremia	NN	O	B-Disease
(	NN	O	O
CHM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
progressive	NN	O	I-Disease
degeneration	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
choroid	NN	O	I-Disease
and	NN	O	I-Disease
retina	NN	O	I-Disease
.	NN	O	O

12	NN	O	O
%	NN	O	O
of	NN	O	O
unrelated	NN	O	O
male	NN	O	O
patients	NN	O	O
carry	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
partially	NN	O	O
cloned	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

In	NN	O	O
Finland	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
more	NN	O	O
than	NN	O	O
120	NN	O	O
living	NN	O	O
CHM	NN	O	B-Disease
patients	NN	O	O
belonging	NN	O	O
to	NN	O	O
eight	NN	O	O
apparently	NN	O	O
unrelated	NN	O	O
pedigrees	NN	O	O
.	NN	O	O

Molecular	NN	O	O
deletions	NN	O	O
involving	NN	O	O
the	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
three	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
screened	NN	O	O
the	NN	O	O
remaining	NN	O	O
five	NN	O	O
families	NN	O	O
for	NN	O	O
point	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
large	NN	O	O
family	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
(	NN	O	O
T	NN	O	O
)	NN	O	O
insertion	NN	O	O
into	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
exon	NN	O	O
C	NN	O	O
leads	NN	O	O
to	NN	O	O
two	NN	O	O
aberrantly	NN	O	O
spliced	NN	O	O
mRNAs	NN	O	O
both	NN	O	O
producing	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
can	NN	O	O
be	NN	O	O
assayed	NN	O	O
easily	NN	O	O
by	NN	O	O
amplification	NN	O	O
and	NN	O	O
digestion	NN	O	O
with	NN	O	O
Msel	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
provide	NN	O	O
additional	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
pathogenetic	NN	O	O
role	NN	O	O
of	NN	O	O
CHM	NN	O	B-Disease
mutations	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
diagnostic	NN	O	O
tool	NN	O	O
for	NN	O	O
one	NN	O	O
fifth	NN	O	O
of	NN	O	O
the	NN	O	O
worlds	NN	O	O
known	NN	O	O
CHM	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Germline	NN	O	O
intronic	NN	O	O
and	NN	O	O
exonic	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
'	NN	O	I-Disease
tumour	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
WT1	NN	O	O
)	NN	O	O
affecting	NN	O	O
urogenital	NN	O	O
development	NN	O	O
.	NN	O	O

Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
human	NN	O	O
developmental	NN	O	B-Disease
disorder	NN	O	I-Disease
affecting	NN	O	O
the	NN	O	O
urogenital	NN	O	O
system	NN	O	O
and	NN	O	O
leading	NN	O	O
to	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
,	NN	O	O
intersex	NN	O	B-Disease
disorders	NN	O	I-Disease
and	NN	O	O
Wilms	NN	O	B-Disease
tumour	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
four	NN	O	O
individuals	NN	O	O
with	NN	O	O
this	NN	O	O
syndrome	NN	O	O
are	NN	O	O
described	NN	O	O
carrying	NN	O	O
germline	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
tumour	NN	O	I-Disease
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
WT1	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
were	NN	O	O
in	NN	O	O
the	NN	O	O
zinc	NN	O	O
finger	NN	O	O
domains	NN	O	O
of	NN	O	O
WT1	NN	O	O
.	NN	O	O

The	NN	O	O
fourth	NN	O	O
occurred	NN	O	O
within	NN	O	O
intron	NN	O	O
9	NN	O	O
,	NN	O	O
preventing	NN	O	O
splicing	NN	O	O
at	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
alternatively	NN	O	O
chosen	NN	O	O
splice	NN	O	O
donor	NN	O	O
sites	NN	O	O
of	NN	O	O
exon	NN	O	O
9	NN	O	O
when	NN	O	O
assayed	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
genetic	NN	O	O
evidence	NN	O	O
for	NN	O	O
distinct	NN	O	O
functional	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
WT1	NN	O	O
isoforms	NN	O	O
in	NN	O	O
urogenital	NN	O	O
development	NN	O	O
.	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
region	NN	O	O
predicts	NN	O	O
multiple	NN	O	O
protein	NN	O	O
isoform	NN	O	O
-	NN	O	O
encoding	NN	O	O
mRNAs	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
underlying	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
polymorphic	NN	O	O
CTG	NN	O	O
-	NN	O	O
repeat	NN	O	O
in	NN	O	O
a	NN	O	O
gene	NN	O	O
encoding	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

Brain	NN	O	O
and	NN	O	O
heart	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	O
-	NN	O	O
kinase	NN	O	O
(	NN	O	O
DMR	NN	O	O
-	NN	O	O
B15	NN	O	O
)	NN	O	O
gene	NN	O	O
are	NN	O	O
subject	NN	O	O
to	NN	O	O
alternative	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
in	NN	O	O
both	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
.	NN	O	O

The	NN	O	O
unstable	NN	O	O
[	NN	O	O
CTG	NN	O	O
]	NN	O	O
5	NN	O	O
-	NN	O	O
30	NN	O	O
motif	NN	O	O
is	NN	O	O
found	NN	O	O
uniquely	NN	O	O
in	NN	O	O
humans	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
flanking	NN	O	O
nucleotides	NN	O	O
are	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
mouse	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
region	NN	O	O
of	NN	O	O
both	NN	O	O
species	NN	O	O
reveals	NN	O	O
another	NN	O	O
active	NN	O	O
gene	NN	O	O
(	NN	O	O
DMR	NN	O	O
-	NN	O	O
N9	NN	O	O
)	NN	O	O
in	NN	O	O
close	NN	O	O
proximity	NN	O	O
to	NN	O	O
the	NN	O	O
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

DMR	NN	O	O
-	NN	O	O
N9	NN	O	O
transcripts	NN	O	O
,	NN	O	O
mainly	NN	O	O
expressed	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
testis	NN	O	O
,	NN	O	O
possess	NN	O	O
a	NN	O	O
single	NN	O	O
,	NN	O	O
large	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
its	NN	O	O
protein	NN	O	O
product	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Clinical	NN	O	O
manifestation	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
expanded	NN	O	O
CTG	NN	O	O
-	NN	O	O
repeat	NN	O	O
compromising	NN	O	O
the	NN	O	O
(	NN	O	O
alternative	NN	O	O
)	NN	O	O
expression	NN	O	O
of	NN	O	O
DM	NN	O	O
-	NN	O	O
kinase	NN	O	O
or	NN	O	O
DMR	NN	O	O
-	NN	O	O
N9	NN	O	O
proteins	NN	O	O
.	NN	O	O
.	NN	O	O

Fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
without	NN	O	O
CCG	NN	O	O
amplification	NN	O	O
has	NN	O	O
an	NN	O	O
FMR1	NN	O	O
deletion	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
typical	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
without	NN	O	O
cytogenetic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
or	NN	O	O
an	NN	O	O
amplified	NN	O	O
CCG	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
fragment	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
has	NN	O	O
a	NN	O	O
previously	NN	O	O
uncharacterized	NN	O	O
submicroscopic	NN	O	O
deletion	NN	O	O
encompassing	NN	O	O
the	NN	O	O
CCG	NN	O	O
repeat	NN	O	O
,	NN	O	O
the	NN	O	O
entire	NN	O	O
FMR1	NN	O	O
gene	NN	O	O
and	NN	O	O
about	NN	O	O
2	NN	O	O
.	NN	O	O

5	NN	O	O
megabases	NN	O	O
of	NN	O	O
flanking	NN	O	O
sequences	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
confirms	NN	O	O
that	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
phenotype	NN	O	O
can	NN	O	O
exist	NN	O	O
,	NN	O	O
without	NN	O	O
amplification	NN	O	O
of	NN	O	O
the	NN	O	O
CCG	NN	O	O
repeat	NN	O	O
or	NN	O	O
cytogenetic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
that	NN	O	O
fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
genetically	NN	O	O
homogeneous	NN	O	O
disorder	NN	O	O
involving	NN	O	O
FMR1	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
random	NN	O	O
X	NN	O	O
-	NN	O	O
inactivation	NN	O	O
in	NN	O	O
the	NN	O	O
mother	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
who	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
carrier	NN	O	O
of	NN	O	O
this	NN	O	O
deletion	NN	O	O
.	NN	O	O

Cloning	NN	O	O
of	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
region	NN	O	O
in	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
2	NN	O	O
.	NN	O	O

5	NN	O	O
million	NN	O	O
base	NN	O	O
pair	NN	O	O
(	NN	O	O
Mb	NN	O	O
)	NN	O	O
region	NN	O	O
between	NN	O	O
the	NN	O	O
loci	NN	O	O
D4S10	NN	O	O
and	NN	O	O
D4S168	NN	O	O
on	NN	O	O
the	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
4	NN	O	O
.	NN	O	O

As	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
strategy	NN	O	O
to	NN	O	O
clone	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
its	NN	O	O
chromosomal	NN	O	O
location	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
region	NN	O	O
as	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
overlapping	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosome	NN	O	O
(	NN	O	O
YAC	NN	O	O
)	NN	O	O
clones	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
eight	NN	O	O
YAC	NN	O	O
clones	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
screening	NN	O	O
human	NN	O	O
YAC	NN	O	O
libraries	NN	O	O
with	NN	O	O
twelve	NN	O	O
PCR	NN	O	O
-	NN	O	O
based	NN	O	O
sequence	NN	O	O
-	NN	O	O
tagged	NN	O	O
sites	NN	O	O
(	NN	O	O
STSs	NN	O	O
)	NN	O	O
from	NN	O	O
the	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
assembled	NN	O	O
the	NN	O	O
YAC	NN	O	O
clones	NN	O	O
into	NN	O	O
overlapping	NN	O	O
sets	NN	O	O
by	NN	O	O
hybridizing	NN	O	O
them	NN	O	O
to	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
DNA	NN	O	O
probes	NN	O	O
from	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
region	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
STSs	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
the	NN	O	O
ends	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
DNA	NN	O	O
inserts	NN	O	O
of	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
YAC	NN	O	O
clones	NN	O	O
to	NN	O	O
assist	NN	O	O
in	NN	O	O
the	NN	O	O
construction	NN	O	O
of	NN	O	O
the	NN	O	O
contig	NN	O	O
.	NN	O	O

Although	NN	O	O
almost	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
YACs	NN	O	O
appear	NN	O	O
to	NN	O	O
contain	NN	O	O
chimeric	NN	O	O
inserts	NN	O	O
and	NN	O	O
several	NN	O	O
contain	NN	O	O
internal	NN	O	O
deletions	NN	O	O
or	NN	O	O
other	NN	O	O
rearrangements	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
obtain	NN	O	O
over	NN	O	O
2	NN	O	O
.	NN	O	O

2	NN	O	O
Mb	NN	O	O
of	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
region	NN	O	O
in	NN	O	O
YACs	NN	O	O
,	NN	O	O
including	NN	O	O
one	NN	O	O
continuous	NN	O	O
segment	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

0	NN	O	O
Mb	NN	O	O
covering	NN	O	O
the	NN	O	O
region	NN	O	O
that	NN	O	O
most	NN	O	O
likely	NN	O	O
contains	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Ten	NN	O	O
of	NN	O	O
the	NN	O	O
twenty	NN	O	O
eight	NN	O	O
YAC	NN	O	O
clones	NN	O	O
comprise	NN	O	O
a	NN	O	O
minimal	NN	O	O
set	NN	O	O
spanning	NN	O	O
the	NN	O	O
2	NN	O	O
.	NN	O	O

2	NN	O	O
Mb	NN	O	O
.	NN	O	O

These	NN	O	O
clones	NN	O	O
provide	NN	O	O
reagents	NN	O	O
for	NN	O	O
the	NN	O	O
complete	NN	O	O
characterization	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
genome	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
eventual	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
YAC	NN	O	O
containing	NN	O	O
part	NN	O	O
or	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
locus	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
from	NN	O	O
pulsed	NN	O	O
-	NN	O	O
field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
PFGE	NN	O	O
)	NN	O	O
that	NN	O	O
the	NN	O	O
monoamine	NN	O	O
oxidase	NN	O	O
genes	NN	O	O
A	NN	O	O
and	NN	O	O
B	NN	O	O
(	NN	O	O
MAOA	NN	O	O
&	NN	O	O
MAOB	NN	O	O
)	NN	O	O
and	NN	O	O
DXS7	NN	O	O
loci	NN	O	O
are	NN	O	O
physically	NN	O	O
very	NN	O	O
close	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
therefore	NN	O	O
extended	NN	O	O
studies	NN	O	O
on	NN	O	O
their	NN	O	O
relationship	NN	O	O
through	NN	O	O
the	NN	O	O
characterisation	NN	O	O
of	NN	O	O
a	NN	O	O
650	NN	O	O
kb	NN	O	O
YAC	NN	O	O
isolated	NN	O	O
using	NN	O	O
L1	NN	O	O
.	NN	O	O

28	NN	O	O
(	NN	O	O
recognising	NN	O	O
the	NN	O	O
DXS7	NN	O	O
locus	NN	O	O
)	NN	O	O
as	NN	O	O
a	NN	O	O
probe	NN	O	O
.	NN	O	O

Restriction	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
YAC	NN	O	O
indicates	NN	O	O
that	NN	O	O
it	NN	O	O
contains	NN	O	O
both	NN	O	O
MAOA	NN	O	O
and	NN	O	O
MAOB	NN	O	O
genes	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
DXS7	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
map	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
YL1	NN	O	O
.	NN	O	O

28	NN	O	O
-	NN	O	O
YAC	NN	O	O
is	NN	O	O
compatible	NN	O	O
both	NN	O	O
with	NN	O	O
the	NN	O	O
map	NN	O	O
from	NN	O	O
an	NN	O	O
independently	NN	O	O
derived	NN	O	O
YAC	NN	O	O
carrying	NN	O	O
MAOA	NN	O	O
and	NN	O	O
B	NN	O	O
genes	NN	O	O
and	NN	O	O
with	NN	O	O
the	NN	O	O
long	NN	O	O
range	NN	O	O
genomic	NN	O	O
map	NN	O	O
for	NN	O	O
the	NN	O	O
region	NN	O	O
.	NN	O	O

A	NN	O	O
series	NN	O	O
of	NN	O	O
subclones	NN	O	O
prepared	NN	O	O
from	NN	O	O
a	NN	O	O
phage	NN	O	O
library	NN	O	O
(	NN	O	O
lambda	NN	O	O
DASH	NN	O	O
II	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
YAC	NN	O	O
have	NN	O	O
been	NN	O	O
characterised	NN	O	O
and	NN	O	O
have	NN	O	O
been	NN	O	O
employed	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
end	NN	O	O
point	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
a	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
NDP	NN	O	B-Disease
)	NN	O	O
patient	NN	O	O
who	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
lack	NN	O	O
both	NN	O	O
DXS7	NN	O	O
and	NN	O	O
MAO	NN	O	O
coding	NN	O	O
sequences	NN	O	O
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
retention	NN	O	O
of	NN	O	O
subclones	NN	O	O
in	NN	O	O
the	NN	O	O
deletion	NN	O	O
patient	NN	O	O
place	NN	O	O
the	NN	O	O
end	NN	O	O
point	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
within	NN	O	O
30	NN	O	O
-	NN	O	O
130	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
YAC	NN	O	O
.	NN	O	O

By	NN	O	O
combining	NN	O	O
the	NN	O	O
data	NN	O	O
with	NN	O	O
established	NN	O	O
recombination	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
all	NN	O	O
or	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
NDP	NN	O	O
lies	NN	O	O
in	NN	O	O
the	NN	O	O
interval	NN	O	O
of	NN	O	O
approximately	NN	O	O
250kb	NN	O	O
within	NN	O	O
the	NN	O	O
YAC	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
in	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
are	NN	O	O
necessary	NN	O	O
to	NN	O	O
produce	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
deficient	NN	O	I-Disease
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
African	NN	O	O
populations	NN	O	O
is	NN	O	O
due	NN	O	O
almost	NN	O	O
entirely	NN	O	O
to	NN	O	O
the	NN	O	O
enzyme	NN	O	O
variant	NN	O	O
A	NN	O	O
-	NN	O	O
,	NN	O	O
which	NN	O	O
differs	NN	O	O
from	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
G6PD	NN	O	O
B	NN	O	O
by	NN	O	O
two	NN	O	O
amino	NN	O	O
acid	NN	O	O
replacements	NN	O	O
,	NN	O	O
68	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
and	NN	O	O
126	NN	O	O
Asn	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Asp	NN	O	O
.	NN	O	O

The	NN	O	O
non	NN	O	O
-	NN	O	O
deficient	NN	O	O
polymorphic	NN	O	O
variant	NN	O	O
G6PD	NN	O	O
A	NN	O	O
contains	NN	O	O
only	NN	O	O
the	NN	O	O
mutation	NN	O	O
126	NN	O	O
Asn	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Asp	NN	O	O
.	NN	O	O

The	NN	O	O
frequencies	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	O
A	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
genes	NN	O	O
in	NN	O	O
parts	NN	O	O
of	NN	O	O
Africa	NN	O	O
are	NN	O	O
both	NN	O	O
about	NN	O	O
0	NN	O	O
.	NN	O	O

2	NN	O	O
.	NN	O	O

The	NN	O	O
68	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
mutation	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
B	NN	O	O
background	NN	O	O
.	NN	O	O

This	NN	O	O
could	NN	O	O
be	NN	O	O
because	NN	O	O
the	NN	O	O
68	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
mutation	NN	O	O
happened	NN	O	O
to	NN	O	O
arise	NN	O	O
in	NN	O	O
an	NN	O	O
A	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
instance	NN	O	O
,	NN	O	O
or	NN	O	O
because	NN	O	O
the	NN	O	O
68	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
mutation	NN	O	O
alone	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
cause	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
approached	NN	O	O
this	NN	O	O
question	NN	O	O
by	NN	O	O
producing	NN	O	O
G6PD	NN	O	O
B	NN	O	O
,	NN	O	O
A	NN	O	O
,	NN	O	O
A	NN	O	O
-	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
68	NN	O	O
Val	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
in	NN	O	O
a	NN	O	O
bacterial	NN	O	O
expression	NN	O	O
system	NN	O	O
and	NN	O	O
analysing	NN	O	O
their	NN	O	O
biochemical	NN	O	O
properties	NN	O	O
.	NN	O	O

With	NN	O	O
each	NN	O	O
single	NN	O	O
mutation	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
slight	NN	O	O
decrease	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
specific	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
yield	NN	O	O
of	NN	O	O
enzyme	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
G6PD	NN	O	O
B	NN	O	O
.	NN	O	O

When	NN	O	O
both	NN	O	O
mutations	NN	O	O
were	NN	O	O
introduced	NN	O	O
together	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
roughly	NN	O	O
additive	NN	O	O
effect	NN	O	O
on	NN	O	O
specific	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
a	NN	O	O
much	NN	O	O
more	NN	O	O
drastic	NN	O	O
effect	NN	O	O
on	NN	O	O
enzyme	NN	O	O
yield	NN	O	O
(	NN	O	O
4	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
on	NN	O	O
thermal	NN	O	O
stability	NN	O	O
,	NN	O	O
especially	NN	O	O
at	NN	O	O
low	NN	O	O
NADP	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Comparable	NN	O	O
results	NN	O	O
were	NN	O	O
produced	NN	O	O
when	NN	O	O
the	NN	O	O
replacement	NN	O	O
119	NN	O	O
Gln	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Glu	NN	O	O
was	NN	O	O
studied	NN	O	O
instead	NN	O	O
of	NN	O	O
126	NN	O	O
Asn	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Asp	NN	O	O
.	NN	O	O

We	NN	O	O
infer	NN	O	O
that	NN	O	O
the	NN	O	O
coexistence	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	I-Disease
G6PD	NN	O	I-Disease
A	NN	O	O
-	NN	O	O
because	NN	O	O
they	NN	O	O
act	NN	O	O
synergistically	NN	O	O
in	NN	O	O
causing	NN	O	O
instability	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
.	NN	O	O

Small	NN	O	O
nuclear	NN	O	O
ribonucleoprotein	NN	O	O
polypeptide	NN	O	O
N	NN	O	O
(	NN	O	O
SNRPN	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
expressed	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
critical	NN	O	O
region	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
paternally	NN	O	O
derived	NN	O	O
chromosomal	NN	O	O
deletions	NN	O	O
in	NN	O	O
region	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
or	NN	O	O
with	NN	O	O
maternal	NN	O	B-Disease
disomy	NN	O	I-Disease
for	NN	O	I-Disease
chromosome	NN	O	I-Disease
15	NN	O	I-Disease
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
expressed	NN	O	O
paternal	NN	O	O
alleles	NN	O	O
of	NN	O	O
maternally	NN	O	O
imprinted	NN	O	O
genes	NN	O	O
must	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
mapped	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
small	NN	O	O
nuclear	NN	O	O
RNA	NN	O	O
associated	NN	O	O
polypeptide	NN	O	O
SmN	NN	O	O
(	NN	O	O
SNRPN	NN	O	O
)	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
15q12	NN	O	O
and	NN	O	O
a	NN	O	O
processed	NN	O	O
pseudogene	NN	O	O
SNRPNP1	NN	O	O
to	NN	O	O
chromosome	NN	O	O
region	NN	O	O
6pter	NN	O	O
-	NN	O	O
p21	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
SNRPN	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
minimal	NN	O	O
deletion	NN	O	O
interval	NN	O	O
that	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
fact	NN	O	O
that	NN	O	O
the	NN	O	O
mouse	NN	O	O
Snrpn	NN	O	O
gene	NN	O	O
is	NN	O	O
maternally	NN	O	O
imprinted	NN	O	O
in	NN	O	O
brain	NN	O	O
suggests	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
paternally	NN	O	O
derived	NN	O	O
SNRPN	NN	O	O
allele	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
infantile	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
Cajun	NN	O	O
population	NN	O	O
.	NN	O	O

A	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
mutation	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
a	NN	O	O
Cajun	NN	O	O
population	NN	O	O
in	NN	O	O
southwest	NN	O	O
Louisiana	NN	O	O
and	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
origins	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
.	NN	O	O

Eleven	NN	O	O
of	NN	O	O
12	NN	O	O
infantile	NN	O	O
TSD	NN	O	B-Disease
alleles	NN	O	O
examined	NN	O	O
in	NN	O	O
six	NN	O	O
families	NN	O	O
had	NN	O	O
the	NN	O	O
beta	NN	O	O
-	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
(	NN	O	O
Hex	NN	O	O
A	NN	O	O
)	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
exon	NN	O	O
11	NN	O	O
insertion	NN	O	O
mutation	NN	O	O
that	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
approximately	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
TSD	NN	O	B-Disease
heterozygotes	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
allele	NN	O	O
was	NN	O	O
a	NN	O	O
single	NN	O	O
-	NN	O	O
base	NN	O	O
transition	NN	O	O
in	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
intron	NN	O	O
9	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
origins	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Cajun	NN	O	O
population	NN	O	O
,	NN	O	O
the	NN	O	O
TSD	NN	O	B-Disease
carrier	NN	O	O
status	NN	O	O
was	NN	O	O
enzymatically	NN	O	O
determined	NN	O	O
for	NN	O	O
90	NN	O	O
members	NN	O	O
of	NN	O	O
four	NN	O	O
of	NN	O	O
the	NN	O	O
six	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
extensive	NN	O	O
pedigrees	NN	O	O
were	NN	O	O
constructed	NN	O	O
for	NN	O	O
all	NN	O	O
carriers	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
ancestral	NN	O	O
couple	NN	O	O
from	NN	O	O
France	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
common	NN	O	O
to	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
exon	NN	O	O
11	NN	O	O
insertion	NN	O	O
.	NN	O	O

Pedigree	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
in	NN	O	O
the	NN	O	O
Cajun	NN	O	O
population	NN	O	O
since	NN	O	O
its	NN	O	O
founding	NN	O	O
over	NN	O	O
2	NN	O	O
centuries	NN	O	O
ago	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
widely	NN	O	O
distributed	NN	O	O
within	NN	O	O
the	NN	O	O
population	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
intron	NN	O	O
9	NN	O	O
mutation	NN	O	O
apparently	NN	O	O
was	NN	O	O
introduced	NN	O	O
within	NN	O	O
the	NN	O	O
last	NN	O	O
century	NN	O	O
and	NN	O	O
probably	NN	O	O
is	NN	O	O
limited	NN	O	O
to	NN	O	O
a	NN	O	O
few	NN	O	O
Louisiana	NN	O	O
families	NN	O	O
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
isolated	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
linked	NN	O	I-Disease
Norrie	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
ND	NN	O	B-Disease
)	NN	O	O
which	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
deleted	NN	O	O
or	NN	O	O
disrupted	NN	O	O
in	NN	O	O
several	NN	O	O
patients	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ND	NN	O	B-Disease
gene	NN	O	O
we	NN	O	O
have	NN	O	O
established	NN	O	O
the	NN	O	O
exon	NN	O	O
-	NN	O	O
intron	NN	O	O
structure	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
17	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
and	NN	O	O
15	NN	O	O
controls	NN	O	O
,	NN	O	O
PCR	NN	O	O
products	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
promoter	NN	O	O
region	NN	O	O
,	NN	O	O
exons	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
coding	NN	O	O
part	NN	O	O
of	NN	O	O
exon	NN	O	O
3	NN	O	O
were	NN	O	O
analysed	NN	O	O
with	NN	O	O
the	NN	O	O
single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
technique	NN	O	O
.	NN	O	O

In	NN	O	O
12	NN	O	O
patients	NN	O	O
altered	NN	O	O
PCR	NN	O	O
fragments	NN	O	O
were	NN	O	O
detected	NN	O	O
which	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
detail	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Eleven	NN	O	O
different	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
but	NN	O	O
one	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
significant	NN	O	O
structural	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
predicted	NN	O	O
protein	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
functionally	NN	O	O
relevant	NN	O	O
base	NN	O	O
changes	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
,	NN	O	O
emphasize	NN	O	O
the	NN	O	O
causal	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
candidate	NN	O	O
gene	NN	O	O
in	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
pave	NN	O	O
the	NN	O	O
way	NN	O	O
for	NN	O	O
reliable	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
carrier	NN	O	O
detection	NN	O	O
.	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
by	NN	O	O
multiple	NN	O	O
SSCP	NN	O	O
.	NN	O	O

A	NN	O	O
combination	NN	O	O
of	NN	O	O
multiplex	NN	O	O
PCR	NN	O	O
with	NN	O	O
the	NN	O	O
single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
technique	NN	O	O
was	NN	O	O
employed	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
.	NN	O	O

Co	NN	O	O
-	NN	O	O
amplification	NN	O	O
of	NN	O	O
11	NN	O	O
exons	NN	O	O
from	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
patients	NN	O	O
with	NN	O	O
no	NN	O	O
deletion	NN	O	O
or	NN	O	O
duplication	NN	O	O
was	NN	O	O
performed	NN	O	O
and	NN	O	O
the	NN	O	O
samples	NN	O	O
subjected	NN	O	O
to	NN	O	O
multiple	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
Duchenne	NN	O	B-Disease
patient	NN	O	O
identified	NN	O	O
by	NN	O	O
this	NN	O	O
approach	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
introduces	NN	O	O
a	NN	O	O
termination	NN	O	O
codon	NN	O	O
within	NN	O	O
exon	NN	O	O
8	NN	O	O
of	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
cause	NN	O	O
a	NN	O	O
very	NN	O	O
premature	NN	O	O
translational	NN	O	O
termination	NN	O	O
accounting	NN	O	O
for	NN	O	O
the	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
inherited	NN	O	O
this	NN	O	O
mutation	NN	O	O
from	NN	O	O
his	NN	O	O
mother	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
the	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
5	NN	O	O
polymorphisms	NN	O	O
useful	NN	O	O
for	NN	O	O
internal	NN	O	O
control	NN	O	O
.	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
-	NN	O	O
related	NN	O	O
expression	NN	O	O
of	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
nucleoside	NN	O	O
diphosphate	NN	O	O
kinase	NN	O	O
.	NN	O	O

High	NN	O	O
level	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
nm23	NN	O	O
-	NN	O	O
H1	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
for	NN	O	O
a	NN	O	O
nucleoside	NN	O	O
diphosphate	NN	O	O
kinase	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
diminished	NN	O	O
metastasis	NN	O	O
in	NN	O	O
some	NN	O	O
tumors	NN	O	B-Disease
but	NN	O	O
not	NN	O	O
in	NN	O	O
others	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
nm23	NN	O	O
-	NN	O	O
H1	NN	O	O
gene	NN	O	O
in	NN	O	O
two	NN	O	O
-	NN	O	O
dimensional	NN	O	O
electrophoretic	NN	O	O
gels	NN	O	O
and	NN	O	O
have	NN	O	O
designated	NN	O	O
it	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
.	NN	O	O

In	NN	O	O
neuroblastoma	NN	O	B-Disease
,	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
amplification	NN	O	O
of	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
myc	NN	O	O
oncogene	NN	O	O
,	NN	O	O
large	NN	O	O
tumor	NN	O	B-Disease
mass	NN	O	O
,	NN	O	O
and	NN	O	O
metastasis	NN	O	O
,	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
advanced	NN	O	O
stage	NN	O	O
tumors	NN	O	B-Disease
compared	NN	O	O
with	NN	O	O
limited	NN	O	O
stage	NN	O	O
disease	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
variable	NN	O	O
expression	NN	O	O
of	NN	O	O
nm23	NN	O	O
-	NN	O	O
H1	NN	O	O
in	NN	O	O
different	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
and	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
were	NN	O	O
compared	NN	O	O
between	NN	O	O
resting	NN	O	O
and	NN	O	O
mitotically	NN	O	O
stimulated	NN	O	O
normal	NN	O	O
human	NN	O	O
PBLs	NN	O	O
and	NN	O	O
in	NN	O	O
leukemia	NN	O	B-Disease
cells	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
increased	NN	O	O
in	NN	O	O
normal	NN	O	O
lymphocytes	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitotic	NN	O	O
stimulation	NN	O	O
and	NN	O	O
paralleled	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
.	NN	O	O

In	NN	O	O
leukemia	NN	O	B-Disease
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
different	NN	O	O
subtypes	NN	O	O
of	NN	O	O
acute	NN	O	B-Disease
leukemia	NN	O	I-Disease
,	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
levels	NN	O	O
were	NN	O	O
also	NN	O	O
increased	NN	O	O
relative	NN	O	O
to	NN	O	O
resting	NN	O	O
normal	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
mitotically	NN	O	O
stimulated	NN	O	O
lymphocytes	NN	O	O
with	NN	O	O
cyclosporin	NN	O	O
,	NN	O	O
which	NN	O	O
inhibits	NN	O	O
proliferation	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
;	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
leukemia	NN	O	B-Disease
cell	NN	O	O
line	NN	O	O
HL	NN	O	O
-	NN	O	O
60	NN	O	O
with	NN	O	O
dimethylsulfoxide	NN	O	O
,	NN	O	O
which	NN	O	O
induces	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
diminished	NN	O	O
levels	NN	O	O
of	NN	O	O
p19	NN	O	O
/	NN	O	O
nm23	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
therefore	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
nm23	NN	O	O
-	NN	O	O
H1	NN	O	O
expression	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
cell	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
.	NN	O	O
.	NN	O	O

An	NN	O	O
intrachromosomal	NN	O	O
insertion	NN	O	O
causing	NN	O	O
5q22	NN	O	O
deletion	NN	O	O
and	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
in	NN	O	O
two	NN	O	O
generations	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
FAPC	NN	O	B-Disease
)	NN	O	O
with	NN	O	O
epidermoid	NN	O	B-Disease
cysts	NN	O	I-Disease
,	NN	O	O
osteomata	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
areas	NN	O	O
of	NN	O	O
congenital	NN	O	B-Disease
hypertrophy	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
retinal	NN	O	I-Disease
pigment	NN	O	I-Disease
epithelium	NN	O	I-Disease
(	NN	O	O
CHRPEs	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
a	NN	O	O
male	NN	O	O
patient	NN	O	O
and	NN	O	O
his	NN	O	O
maternal	NN	O	O
aunt	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
whom	NN	O	O
suffered	NN	O	O
a	NN	O	O
mild	NN	O	O
to	NN	O	O
moderate	NN	O	O
degree	NN	O	O
of	NN	O	O
mental	NN	O	B-Disease
handicap	NN	O	I-Disease
.	NN	O	O

Both	NN	O	O
had	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
(	NN	O	O
del	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
(	NN	O	O
q22q23	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
normal	NN	O	O
family	NN	O	O
members	NN	O	O
had	NN	O	O
the	NN	O	O
underlying	NN	O	O
direct	NN	O	O
insertion	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
(	NN	O	O
dir	NN	O	O
ins	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
(	NN	O	O
q31	NN	O	O
.	NN	O	O
3q22q23	NN	O	O
3q22q23	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
.	NN	O	O

Molecular	NN	O	O
genetic	NN	O	O
and	NN	O	O
fluorescent	NN	O	O
hybridisation	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
loci	NN	O	O
D5S37	NN	O	O
and	NN	O	O
D5S98	NN	O	O
are	NN	O	O
outside	NN	O	O
the	NN	O	O
deletion	NN	O	O
whereas	NN	O	O
loci	NN	O	O
detected	NN	O	O
by	NN	O	O
probes	NN	O	O
EF5	NN	O	O
.	NN	O	O

44	NN	O	O
and	NN	O	O
YN5	NN	O	O
.	NN	O	O

48	NN	O	O
are	NN	O	O
lost	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
loss	NN	O	O
of	NN	O	O
one	NN	O	O
allele	NN	O	O
at	NN	O	O
the	NN	O	O
MCC	NN	O	O
and	NN	O	O
APC	NN	O	B-Disease
loci	NN	O	O
.	NN	O	O

The	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
band	NN	O	O
5q22	NN	O	O
.	NN	O	O

Familial	NN	O	O
direct	NN	O	O
insertions	NN	O	O
should	NN	O	O
be	NN	O	O
considered	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
recurrent	NN	O	O
microdeletion	NN	O	O
syndromes	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
rod	NN	O	O
photoreceptor	NN	O	O
cGMP	NN	O	O
phosphodiesterase	NN	O	O
beta	NN	O	O
-	NN	O	O
subunit	NN	O	O
gene	NN	O	O
in	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
:	NN	O	O
tight	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
region	NN	O	O
(	NN	O	O
4p16	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
mouse	NN	O	O
(	NN	O	O
gene	NN	O	O
symbol	NN	O	O
,	NN	O	O
rd	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
certain	NN	O	O
forms	NN	O	O
of	NN	O	O
human	NN	O	O
hereditary	NN	O	B-Disease
retinopathies	NN	O	I-Disease
.	NN	O	O

Recent	NN	O	O
findings	NN	O	O
of	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
rd	NN	O	O
mouse	NN	O	O
PDE	NN	O	O
beta	NN	O	O
-	NN	O	O
subunit	NN	O	O
gene	NN	O	O
(	NN	O	O
Pdeb	NN	O	O
)	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
chromosome	NN	O	O
locations	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
utilized	NN	O	O
backcross	NN	O	O
analysis	NN	O	O
in	NN	O	O
mice	NN	O	O
to	NN	O	O
verify	NN	O	O
and	NN	O	O
define	NN	O	O
more	NN	O	O
precisely	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
Pdeb	NN	O	O
locus	NN	O	O
6	NN	O	O
.	NN	O	O

1	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O

3	NN	O	O
cM	NN	O	O
distal	NN	O	O
of	NN	O	O
Mgsa	NN	O	O
on	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
5	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
(	NN	O	O
PDEB	NN	O	O
)	NN	O	O
maps	NN	O	O
to	NN	O	O
4p16	NN	O	O
.	NN	O	O

3	NN	O	O
,	NN	O	O
very	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
region	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
comparative	NN	O	O
map	NN	O	O
for	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
will	NN	O	O
reside	NN	O	O
on	NN	O	O
chromosome	NN	O	O
5	NN	O	O
.	NN	O	O

Linkage	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
Pdeb	NN	O	O
locus	NN	O	O
with	NN	O	O
other	NN	O	O
homologues	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
4p16	NN	O	O
.	NN	O	O

3	NN	O	O
region	NN	O	O
is	NN	O	O
maintained	NN	O	O
but	NN	O	O
gene	NN	O	O
order	NN	O	O
is	NN	O	O
not	NN	O	O
,	NN	O	O
suggesting	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
possible	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
corresponding	NN	O	O
mouse	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Coincident	NN	O	O
Kaposi	NN	O	B-Disease
sarcoma	NN	O	I-Disease
and	NN	O	O
T	NN	O	B-Disease
-	NN	O	I-Disease
cell	NN	O	I-Disease
lymphoma	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
24	NN	O	O
year	NN	O	O
old	NN	O	O
male	NN	O	O
with	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
eczema	NN	O	B-Disease
,	NN	O	O
recurrent	NN	O	O
mild	NN	O	O
infections	NN	O	O
,	NN	O	O
and	NN	O	O
thrombocytopenia	NN	O	B-Disease
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
presented	NN	O	O
with	NN	O	O
a	NN	O	O
mediastinal	NN	O	O
mass	NN	O	O
,	NN	O	O
generalized	NN	O	O
lymphadenopathy	NN	O	B-Disease
,	NN	O	O
splenomegaly	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
severe	NN	O	O
thrombocytopenia	NN	O	B-Disease
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
immune	NN	O	O
function	NN	O	O
including	NN	O	O
immunoglobulin	NN	O	O
levels	NN	O	O
and	NN	O	O
T	NN	O	O
-	NN	O	O
cell	NN	O	O
subsets	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
his	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
proliferated	NN	O	O
normally	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
phytohemagglutinin	NN	O	O
,	NN	O	O
concanavalin	NN	O	O
A	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
neuraminidase	NN	O	O
/	NN	O	O
galactose	NN	O	O
oxidase	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
their	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
to	NN	O	O
anti	NN	O	O
-	NN	O	O
CD43	NN	O	O
antibody	NN	O	O
and	NN	O	O
periodate	NN	O	O
were	NN	O	O
diminished	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
clinical	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O

An	NN	O	O
initial	NN	O	O
inguinal	NN	O	O
lymph	NN	O	O
node	NN	O	O
biopsy	NN	O	O
surprisingly	NN	O	O
revealed	NN	O	O
Kaposi	NN	O	B-Disease
sarcoma	NN	O	I-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
following	NN	O	O
splenectomy	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
platelet	NN	O	O
count	NN	O	O
,	NN	O	O
biopsy	NN	O	O
of	NN	O	O
the	NN	O	O
mediastinal	NN	O	O
mass	NN	O	O
revealed	NN	O	O
T	NN	O	B-Disease
-	NN	O	I-Disease
cell	NN	O	I-Disease
large	NN	O	I-Disease
cell	NN	O	I-Disease
lymphoma	NN	O	I-Disease
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
biopsied	NN	O	O
tissue	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Epstein	NN	O	B-Disease
-	NN	O	I-Disease
Barr	NN	O	I-Disease
virus	NN	O	I-Disease
and	NN	O	O
cytomegalovirus	NN	O	B-Disease
were	NN	O	O
negative	NN	O	O
,	NN	O	O
as	NN	O	O
were	NN	O	O
studies	NN	O	O
of	NN	O	O
blood	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-Disease
immunodeficiency	NN	O	I-Disease
virus	NN	O	I-Disease
(	NN	O	O
HIV	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
Kaposi	NN	O	B-Disease
sarcoma	NN	O	I-Disease
arising	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
congenital	NN	O	B-Disease
immunodeficiency	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
Kaposi	NN	O	B-Disease
sarcoma	NN	O	I-Disease
can	NN	O	O
arise	NN	O	O
in	NN	O	O
the	NN	O	O
face	NN	O	O
of	NN	O	O
the	NN	O	O
severe	NN	O	O
immunosuppression	NN	O	O
that	NN	O	O
follows	NN	O	O
allograft	NN	O	O
transplantation	NN	O	O
and	NN	O	O
in	NN	O	O
patients	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
postulate	NN	O	O
that	NN	O	O
longevity	NN	O	O
in	NN	O	O
the	NN	O	O
face	NN	O	O
of	NN	O	O
mild	NN	O	O
immunosuppression	NN	O	O
was	NN	O	O
the	NN	O	O
major	NN	O	O
factor	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
Kaposi	NN	O	B-Disease
sarcoma	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
responsible	NN	O	O
for	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
mutated	NN	O	O
in	NN	O	O
human	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
cancer	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
world	NN	O	O
,	NN	O	O
genetic	NN	O	O
changes	NN	O	O
during	NN	O	O
its	NN	O	O
carcinogenesis	NN	O	O
are	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
.	NN	O	O

Since	NN	O	O
some	NN	O	O
gastric	NN	O	B-Disease
cancers	NN	O	I-Disease
are	NN	O	O
considered	NN	O	O
to	NN	O	O
originate	NN	O	O
from	NN	O	O
the	NN	O	O
intestinal	NN	O	O
metaplasia	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
mutation	NN	O	O
of	NN	O	O
which	NN	O	O
causes	NN	O	O
adenomatous	NN	O	B-Disease
polyps	NN	O	I-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
colon	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
carcinogenesis	NN	O	O
of	NN	O	O
gastric	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Based	NN	O	O
on	NN	O	O
this	NN	O	O
idea	NN	O	O
,	NN	O	O
DNAs	NN	O	O
isolated	NN	O	O
from	NN	O	O
gastric	NN	O	B-Disease
cancers	NN	O	I-Disease
were	NN	O	O
examined	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
RNase	NN	O	O
protection	NN	O	O
analysis	NN	O	O
coupled	NN	O	O
with	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
followed	NN	O	O
by	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
products	NN	O	O
.	NN	O	O

By	NN	O	O
screening	NN	O	O
nearly	NN	O	O
one	NN	O	O
-	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
44	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
three	NN	O	O
tumors	NN	O	B-Disease
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
5	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
frame	NN	O	O
shift	NN	O	O
which	NN	O	O
causes	NN	O	O
truncation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
product	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
also	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
during	NN	O	O
the	NN	O	O
carcinogenesis	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
gastric	NN	O	B-Disease
cancers	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
71	NN	O	O
-	NN	O	O
kilodalton	NN	O	O
protein	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
other	NN	O	O
nonmuscle	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
known	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
products	NN	O	O
,	NN	O	O
the	NN	O	O
muscle	NN	O	O
-	NN	O	O
and	NN	O	O
brain	NN	O	O
-	NN	O	O
type	NN	O	O
dystrophin	NN	O	O
isoforms	NN	O	O
,	NN	O	O
are	NN	O	O
427	NN	O	O
-	NN	O	O
kDa	NN	O	O
proteins	NN	O	O
translated	NN	O	O
from	NN	O	O
14	NN	O	O
-	NN	O	O
kilobase	NN	O	O
(	NN	O	O
kb	NN	O	O
)	NN	O	O
mRNAs	NN	O	O
.	NN	O	O

Recently	NN	O	O
we	NN	O	O
described	NN	O	O
a	NN	O	O
6	NN	O	O
.	NN	O	O

5	NN	O	O
-	NN	O	O
kb	NN	O	O
mRNA	NN	O	O
that	NN	O	O
also	NN	O	O
is	NN	O	O
transcribed	NN	O	O
from	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Cloning	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
6	NN	O	O
.	NN	O	O

5	NN	O	O
-	NN	O	O
kb	NN	O	O
mRNA	NN	O	O
encodes	NN	O	O
a	NN	O	O
70	NN	O	O
.	NN	O	O

8	NN	O	O
-	NN	O	O
kDa	NN	O	O
protein	NN	O	O
that	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

It	NN	O	O
contains	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
and	NN	O	O
the	NN	O	O
cysteine	NN	O	O
-	NN	O	O
rich	NN	O	O
domains	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
,	NN	O	O
seven	NN	O	O
additional	NN	O	O
amino	NN	O	O
acids	NN	O	O
at	NN	O	O
the	NN	O	O
N	NN	O	O
terminus	NN	O	O
,	NN	O	O
and	NN	O	O
some	NN	O	O
modifications	NN	O	O
formed	NN	O	O
by	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
in	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
domain	NN	O	O
.	NN	O	O

It	NN	O	O
lacks	NN	O	O
the	NN	O	O
entire	NN	O	O
large	NN	O	O
domain	NN	O	O
of	NN	O	O
spectrin	NN	O	O
-	NN	O	O
like	NN	O	O
repeats	NN	O	O
and	NN	O	O
the	NN	O	O
actin	NN	O	O
-	NN	O	O
binding	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
domain	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
.	NN	O	O

This	NN	O	O
protein	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
other	NN	O	O
nonmuscle	NN	O	O
tissues	NN	O	O
but	NN	O	O
is	NN	O	O
undetectable	NN	O	O
in	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
extracts	NN	O	O
.	NN	O	O

Correlation	NN	O	O
between	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
polyps	NN	O	I-Disease
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Recently	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
which	NN	O	O
causes	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
substantial	NN	O	O
number	NN	O	O
of	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
this	NN	O	O
information	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
22	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
(	NN	O	O
12	NN	O	O
of	NN	O	O
whom	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
)	NN	O	O
with	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
polyps	NN	O	I-Disease
developed	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
;	NN	O	O
17	NN	O	O
were	NN	O	O
sparse	NN	O	O
types	NN	O	O
and	NN	O	O
five	NN	O	O
were	NN	O	O
profuse	NN	O	O
types	NN	O	O
.	NN	O	O

All	NN	O	O
but	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
considered	NN	O	O
to	NN	O	O
cause	NN	O	O
truncation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
product	NN	O	O
by	NN	O	O
frame	NN	O	O
-	NN	O	O
shift	NN	O	O
due	NN	O	O
to	NN	O	O
deletion	NN	O	O
(	NN	O	O
14	NN	O	O
cases	NN	O	O
)	NN	O	O
or	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
(	NN	O	O
seven	NN	O	O
cases	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
seems	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
two	NN	O	O
clinical	NN	O	O
types	NN	O	O
;	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
five	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
profuse	NN	O	O
polyps	NN	O	B-Disease
were	NN	O	O
observed	NN	O	O
between	NN	O	O
codon	NN	O	O
1250	NN	O	O
and	NN	O	O
codon	NN	O	O
1464	NN	O	O
,	NN	O	O
whereas	NN	O	O
mutations	NN	O	O
in	NN	O	O
17	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
fewer	NN	O	O
polyps	NN	O	B-Disease
were	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
result	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
polyps	NN	O	I-Disease
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
stability	NN	O	O
or	NN	O	O
biological	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
truncated	NN	O	O
APC	NN	O	B-Disease
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
and	NN	O	O
rapid	NN	O	O
detection	NN	O	O
of	NN	O	O
three	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Moroccan	NN	O	O
Jewish	NN	O	O
population	NN	O	O
.	NN	O	O

Infantile	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
that	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
complete	NN	O	O
absence	NN	O	O
of	NN	O	O
beta	NN	O	O
-	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
activity	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
well	NN	O	O
known	NN	O	O
that	NN	O	O
an	NN	O	O
elevated	NN	O	O
frequency	NN	O	O
of	NN	O	O
TSD	NN	O	B-Disease
mutations	NN	O	O
exists	NN	O	O
among	NN	O	O
Ashkenazi	NN	O	O
Jews	NN	O	O
.	NN	O	O

More	NN	O	O
recently	NN	O	O
it	NN	O	O
has	NN	O	O
become	NN	O	O
apparent	NN	O	O
that	NN	O	O
elevated	NN	O	O
carrier	NN	O	O
frequencies	NN	O	O
for	NN	O	O
TSD	NN	O	B-Disease
also	NN	O	O
occur	NN	O	O
in	NN	O	O
several	NN	O	O
other	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
,	NN	O	O
including	NN	O	O
Moroccan	NN	O	O
Jews	NN	O	O
,	NN	O	O
a	NN	O	O
subgroup	NN	O	O
of	NN	O	O
Sephardic	NN	O	O
Jews	NN	O	O
.	NN	O	O

Elsewhere	NN	O	O
we	NN	O	O
reported	NN	O	O
an	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
adjacent	NN	O	O
phenylalanine	NN	O	O
codons	NN	O	O
at	NN	O	O
position	NN	O	O
304	NN	O	O
or	NN	O	O
305	NN	O	O
(	NN	O	O
delta	NN	O	O
F304	NN	O	O
/	NN	O	O
305	NN	O	O
)	NN	O	O
in	NN	O	O
one	NN	O	O
HEXA	NN	O	O
allele	NN	O	O
of	NN	O	O
a	NN	O	O
Moroccan	NN	O	O
Jewish	NN	O	O
TSD	NN	O	B-Disease
patient	NN	O	O
and	NN	O	O
in	NN	O	O
three	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
from	NN	O	O
six	NN	O	O
unrelated	NN	O	O
Moroccan	NN	O	O
Jewish	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
identified	NN	O	O
two	NN	O	O
additional	NN	O	O
mutations	NN	O	O
within	NN	O	O
exon	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
that	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
remaining	NN	O	O
TSD	NN	O	B-Disease
alleles	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
carriers	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
G	NN	O	O
transversion	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
replacement	NN	O	O
of	NN	O	O
Tyr180	NN	O	O
by	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
Arg170	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
Gln	NN	O	O
substitution	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
is	NN	O	O
at	NN	O	O
a	NN	O	O
CpG	NN	O	O
site	NN	O	O
in	NN	O	O
a	NN	O	O
Japanese	NN	O	O
infant	NN	O	O
with	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
and	NN	O	O
was	NN	O	O
described	NN	O	O
elsewhere	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
nine	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
from	NN	O	O
seven	NN	O	O
unrelated	NN	O	O
families	NN	O	O
showed	NN	O	O
that	NN	O	O
four	NN	O	O
harbor	NN	O	O
the	NN	O	O
delta	NN	O	O
F304	NN	O	O
/	NN	O	O
305	NN	O	O
mutation	NN	O	O
,	NN	O	O
two	NN	O	O
the	NN	O	O
Arg170	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Gln	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
the	NN	O	O
Tyr180	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Stop	NN	O	O
mutation	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
have	NN	O	O
developed	NN	O	O
rapid	NN	O	O
,	NN	O	O
nonradioactive	NN	O	O
assays	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
each	NN	O	O
mutation	NN	O	O
,	NN	O	O
which	NN	O	O
should	NN	O	O
be	NN	O	O
helpful	NN	O	O
for	NN	O	O
carrier	NN	O	O
screening	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
by	NN	O	O
natural	NN	O	O
and	NN	O	O
amplification	NN	O	O
created	NN	O	O
restriction	NN	O	O
sites	NN	O	O
:	NN	O	O
five	NN	O	O
mutations	NN	O	O
account	NN	O	O
for	NN	O	O
most	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
cases	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
simple	NN	O	O
method	NN	O	O
to	NN	O	O
diagnose	NN	O	O
the	NN	O	O
molecular	NN	O	O
defects	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Chinese	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

This	NN	O	O
method	NN	O	O
involves	NN	O	O
the	NN	O	O
selective	NN	O	O
amplification	NN	O	O
of	NN	O	O
a	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
from	NN	O	O
human	NN	O	O
G6PD	NN	O	O
gene	NN	O	O
with	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
primers	NN	O	O
followed	NN	O	O
by	NN	O	O
digestion	NN	O	O
with	NN	O	O
restriction	NN	O	O
enzymes	NN	O	O
that	NN	O	O
recognize	NN	O	O
artificially	NN	O	O
created	NN	O	O
or	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
restriction	NN	O	O
sites	NN	O	O
.	NN	O	O

Ninety	NN	O	O
-	NN	O	O
four	NN	O	O
Chinese	NN	O	O
males	NN	O	O
with	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
50	NN	O	O
%	NN	O	O
(	NN	O	O
47	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
were	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
mutation	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
(	NN	O	O
nt	NN	O	O
)	NN	O	O
1376	NN	O	O
,	NN	O	O
21	NN	O	O
.	NN	O	O

3	NN	O	O
%	NN	O	O
(	NN	O	O
20	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
were	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
1388	NN	O	O
,	NN	O	O
7	NN	O	O
.	NN	O	O

4	NN	O	O
%	NN	O	O
(	NN	O	O
7	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
were	NN	O	O
A	NN	O	O
to	NN	O	O
G	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
493	NN	O	O
,	NN	O	O
7	NN	O	O
.	NN	O	O

4	NN	O	O
%	NN	O	O
(	NN	O	O
7	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
were	NN	O	O
A	NN	O	O
to	NN	O	O
G	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
95	NN	O	O
,	NN	O	O
4	NN	O	O
.	NN	O	O

2	NN	O	O
%	NN	O	O
(	NN	O	O
4	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
were	NN	O	O
C	NN	O	O
to	NN	O	O
T	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
1024	NN	O	O
,	NN	O	O
1	NN	O	O
.	NN	O	O

1	NN	O	O
%	NN	O	O
(	NN	O	O
1	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
was	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
392	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
.	NN	O	O

1	NN	O	O
%	NN	O	O
(	NN	O	O
1	NN	O	O
of	NN	O	O
94	NN	O	O
)	NN	O	O
was	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
mutation	NN	O	O
at	NN	O	O
nt	NN	O	O
487	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
former	NN	O	O
five	NN	O	O
mutations	NN	O	O
account	NN	O	O
for	NN	O	O
more	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
cases	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

Aside	NN	O	O
from	NN	O	O
showing	NN	O	O
that	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
change	NN	O	O
at	NN	O	O
nt	NN	O	O
1376	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
,	NN	O	O
our	NN	O	O
research	NN	O	O
indicates	NN	O	O
that	NN	O	O
nt	NN	O	O
493	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
frequent	NN	O	O
mutation	NN	O	O
among	NN	O	O
Chinese	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
G6PD	NN	O	O
activity	NN	O	O
among	NN	O	O
different	NN	O	O
mutations	NN	O	O
,	NN	O	O
without	NN	O	O
discovering	NN	O	O
significant	NN	O	O
differences	NN	O	O
between	NN	O	O
them	NN	O	O
.	NN	O	O

Eight	NN	O	O
novel	NN	O	O
inactivating	NN	O	O
germ	NN	O	O
line	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
identified	NN	O	O
by	NN	O	O
denaturing	NN	O	O
gradient	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
dominantly	NN	O	O
inherited	NN	O	O
condition	NN	O	O
predisposing	NN	O	O
to	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
recent	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
responsible	NN	O	O
gene	NN	O	O
(	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
or	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
facilitated	NN	O	O
the	NN	O	O
search	NN	O	O
for	NN	O	O
germ	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
affected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Previous	NN	O	O
authors	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
RNase	NN	O	O
protection	NN	O	O
assay	NN	O	O
and	NN	O	O
the	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphisms	NN	O	O
procedure	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
used	NN	O	O
denaturing	NN	O	O
gradient	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
DGGE	NN	O	O
)	NN	O	O
.	NN	O	O

DGGE	NN	O	O
analysis	NN	O	O
of	NN	O	O
10	NN	O	O
APC	NN	O	B-Disease
exons	NN	O	O
(	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8	NN	O	O
,	NN	O	O
9	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
12	NN	O	O
,	NN	O	O
13	NN	O	O
,	NN	O	O
14	NN	O	O
,	NN	O	O
and	NN	O	O
part	NN	O	O
of	NN	O	O
15	NN	O	O
)	NN	O	O
in	NN	O	O
33	NN	O	O
unrelated	NN	O	O
Dutch	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
eight	NN	O	O
novel	NN	O	O
germ	NN	O	O
line	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
stop	NN	O	O
codons	NN	O	O
or	NN	O	O
frameshifts	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
reported	NN	O	O
here	NN	O	O
indicate	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
an	NN	O	O
extremely	NN	O	O
heterogeneous	NN	O	O
spectrum	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
all	NN	O	O
the	NN	O	O
mutations	NN	O	O
found	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
are	NN	O	O
chain	NN	O	O
terminating	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
DGGE	NN	O	O
represents	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
sensitive	NN	O	O
technique	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
unusually	NN	O	O
large	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

An	NN	O	O
extension	NN	O	O
of	NN	O	O
the	NN	O	O
DGGE	NN	O	O
analysis	NN	O	O
to	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
in	NN	O	O
a	NN	O	O
sufficient	NN	O	O
number	NN	O	O
of	NN	O	O
clinically	NN	O	O
well	NN	O	O
-	NN	O	O
characterized	NN	O	O
,	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
will	NN	O	O
facilitate	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
an	NN	O	O
extremely	NN	O	O
heterogeneous	NN	O	O
spectrum	NN	O	O
of	NN	O	O
mutations	NN	O	O
spread	NN	O	O
throughout	NN	O	O
the	NN	O	O
entire	NN	O	O
length	NN	O	O
of	NN	O	O
the	NN	O	O
large	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
among	NN	O	O
the	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
indicates	NN	O	O
that	NN	O	O
this	NN	O	O
approach	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
useful	NN	O	O
as	NN	O	O
a	NN	O	O
rapid	NN	O	O
presymptomatic	NN	O	O
diagnostic	NN	O	O
procedure	NN	O	O
in	NN	O	O
a	NN	O	O
routine	NN	O	O
laboratory	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
the	NN	O	O
above	NN	O	O
DGGE	NN	O	O
approach	NN	O	O
has	NN	O	O
incidentally	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
exon	NN	O	O
13	NN	O	O
.	NN	O	O

Such	NN	O	O
intragenic	NN	O	O
polymorphisms	NN	O	O
offer	NN	O	O
a	NN	O	O
practical	NN	O	O
approach	NN	O	O
to	NN	O	O
a	NN	O	O
more	NN	O	O
rapid	NN	O	O
procedure	NN	O	O
for	NN	O	O
presymptomatic	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
FAP	NN	O	B-Disease
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
informative	NN	O	O
families	NN	O	O
.	NN	O	O
.	NN	O	O

Yeast	NN	O	O
artificial	NN	O	O
chromosomes	NN	O	O
for	NN	O	O
the	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
familial	NN	O	B-Disease
polyposis	NN	O	I-Disease
APC	NN	O	I-Disease
gene	NN	O	O
region	NN	O	O
.	NN	O	O

Two	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosomes	NN	O	O
(	NN	O	O
YACs	NN	O	O
)	NN	O	O
spanning	NN	O	O
a	NN	O	O
total	NN	O	O
distance	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

1	NN	O	O
megabase	NN	O	O
pairs	NN	O	O
of	NN	O	O
DNA	NN	O	O
around	NN	O	O
the	NN	O	O
MCC	NN	O	O
(	NN	O	O
for	NN	O	O
mutated	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
carcinoma	NN	O	I-Disease
)	NN	O	O
and	NN	O	O
APC	NN	O	B-Disease
(	NN	O	O
for	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
)	NN	O	O
genes	NN	O	O
at	NN	O	O
5q21	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
and	NN	O	O
characterized	NN	O	O
.	NN	O	O

Starting	NN	O	O
from	NN	O	O
the	NN	O	O
MCC	NN	O	O
gene	NN	O	O
,	NN	O	O
a	NN	O	O
strategy	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
identify	NN	O	O
constitutional	NN	O	O
submicroscopic	NN	O	O
deletions	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
patients	NN	O	O
that	NN	O	O
might	NN	O	O
considerably	NN	O	O
narrow	NN	O	O
down	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

To	NN	O	O
this	NN	O	O
end	NN	O	O
,	NN	O	O
YACs	NN	O	O
identified	NN	O	O
by	NN	O	O
the	NN	O	O
MCC	NN	O	O
gene	NN	O	O
were	NN	O	O
screened	NN	O	O
across	NN	O	O
a	NN	O	O
chromosome	NN	O	O
5	NN	O	O
-	NN	O	O
specific	NN	O	O
cosmid	NN	O	O
library	NN	O	O
to	NN	O	O
provide	NN	O	O
a	NN	O	O
source	NN	O	O
of	NN	O	O
DNA	NN	O	O
probes	NN	O	O
for	NN	O	O
genomic	NN	O	O
scanning	NN	O	O
.	NN	O	O

The	NN	O	O
cosmids	NN	O	O
isolated	NN	O	O
from	NN	O	O
these	NN	O	O
experiments	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
screen	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
containing	NN	O	O
chromosome	NN	O	O
5	NN	O	O
segregated	NN	O	O
from	NN	O	O
patients	NN	O	O
suspected	NN	O	O
to	NN	O	O
carry	NN	O	O
putative	NN	O	O
interstitial	NN	O	O
deletions	NN	O	O
.	NN	O	O

This	NN	O	O
screening	NN	O	O
approach	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
confirmation	NN	O	O
of	NN	O	O
a	NN	O	O
small	NN	O	O
heterozygous	NN	O	O
deletion	NN	O	O
in	NN	O	O
a	NN	O	O
polyposis	NN	O	B-Disease
patient	NN	O	O
that	NN	O	O
overlaps	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
isolated	NN	O	O
YACs	NN	O	O
.	NN	O	O

This	NN	O	O
YAC	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
the	NN	O	O
entire	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
significant	NN	O	O
portion	NN	O	O
of	NN	O	O
DNA	NN	O	O
flanking	NN	O	O
the	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
should	NN	O	O
therefore	NN	O	O
prove	NN	O	O
a	NN	O	O
valuable	NN	O	O
resource	NN	O	O
for	NN	O	O
functional	NN	O	O
studies	NN	O	O
by	NN	O	O
transfer	NN	O	O
to	NN	O	O
colorectal	NN	O	B-Disease
tumor	NN	O	I-Disease
-	NN	O	O
derived	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

Inherited	NN	O	O
WT1	NN	O	O
mutation	NN	O	O
in	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
the	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
,	NN	O	O
genital	NN	O	B-Disease
anomalies	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
nephropathy	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
carry	NN	O	O
de	NN	O	O
novo	NN	O	O
constitutional	NN	O	O
mutations	NN	O	O
in	NN	O	O
WT1	NN	O	O
,	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
gene	NN	O	O
at	NN	O	O
chromosome	NN	O	O
11p13	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
three	NN	O	O
new	NN	O	O
cases	NN	O	O
,	NN	O	O
two	NN	O	O
carrying	NN	O	O
a	NN	O	O
previously	NN	O	O
described	NN	O	O
WT1	NN	O	O
exon	NN	O	O
9	NN	O	O
mutation	NN	O	O
and	NN	O	O
one	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
WT1	NN	O	O
exon	NN	O	O
8	NN	O	O
mutation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
unlike	NN	O	O
patients	NN	O	O
in	NN	O	O
previous	NN	O	O
reports	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
our	NN	O	O
three	NN	O	O
patients	NN	O	O
inherited	NN	O	O
the	NN	O	O
affected	NN	O	O
allele	NN	O	O
from	NN	O	O
his	NN	O	O
phenotypically	NN	O	O
unaffected	NN	O	O
father	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
WT1	NN	O	O
exon	NN	O	O
9	NN	O	O
mutation	NN	O	O
affecting	NN	O	O
394Arg	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
over	NN	O	O
one	NN	O	O
-	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
may	NN	O	O
exhibit	NN	O	O
incomplete	NN	O	O
penetrance	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
familial	NN	O	O
studies	NN	O	O
in	NN	O	O
patients	NN	O	O
affected	NN	O	O
by	NN	O	O
this	NN	O	O
syndrome	NN	O	O
are	NN	O	O
recommended	NN	O	O
.	NN	O	O
.	NN	O	O

Submicroscopic	NN	O	O
deletions	NN	O	O
at	NN	O	O
the	NN	O	O
WAGR	NN	O	B-Disease
locus	NN	O	O
,	NN	O	O
revealed	NN	O	O
by	NN	O	O
nonradioactive	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
with	NN	O	O
biotin	NN	O	O
-	NN	O	O
labeled	NN	O	O
probes	NN	O	O
mapping	NN	O	O
to	NN	O	O
11p13	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
for	NN	O	O
the	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
WAGR	NN	O	B-Disease
(	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
,	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
genitourinary	NN	O	B-Disease
abnormalities	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
)	NN	O	O
locus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
a	NN	O	O
submicroscopic	NN	O	O
11p13	NN	O	O
deletion	NN	O	O
in	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
inherited	NN	O	O
aniridia	NN	O	B-Disease
who	NN	O	O
subsequently	NN	O	O
presented	NN	O	O
with	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
horseshoe	NN	O	O
kidney	NN	O	O
,	NN	O	O
only	NN	O	O
revealed	NN	O	O
at	NN	O	O
surgery	NN	O	O
.	NN	O	O

The	NN	O	O
mother	NN	O	O
,	NN	O	O
who	NN	O	O
has	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
a	NN	O	O
deletion	NN	O	O
including	NN	O	O
both	NN	O	O
the	NN	O	O
aniridia	NN	O	B-Disease
candidate	NN	O	O
gene	NN	O	O
(	NN	O	O
AN2	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
predisposition	NN	O	O
gene	NN	O	O
(	NN	O	O
WT1	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
therefore	NN	O	O
a	NN	O	O
rare	NN	O	O
case	NN	O	O
of	NN	O	O
an	NN	O	O
inherited	NN	O	O
WAGR	NN	O	B-Disease
deletion	NN	O	O
.	NN	O	O

Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
has	NN	O	O
so	NN	O	O
far	NN	O	O
only	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
sporadic	NN	O	O
de	NN	O	O
novo	NN	O	O
aniridia	NN	O	B-Disease
cases	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
cosmid	NN	O	O
probe	NN	O	O
for	NN	O	O
a	NN	O	O
candidate	NN	O	O
aniridia	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
mouse	NN	O	O
Pax	NN	O	O
-	NN	O	O
6	NN	O	O
gene	NN	O	O
,	NN	O	O
is	NN	O	O
deleted	NN	O	O
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
from	NN	O	O
aniridia	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
previously	NN	O	O
characterized	NN	O	O
deletions	NN	O	O
at	NN	O	O
11p13	NN	O	O
,	NN	O	O
while	NN	O	O
another	NN	O	O
cosmid	NN	O	O
marker	NN	O	O
mapping	NN	O	O
between	NN	O	O
two	NN	O	O
aniridia	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
(	NN	O	O
and	NN	O	O
hence	NN	O	O
a	NN	O	O
second	NN	O	O
candidate	NN	O	O
marker	NN	O	O
)	NN	O	O
is	NN	O	O
present	NN	O	O
on	NN	O	O
both	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
Pax	NN	O	O
-	NN	O	O
6	NN	O	O
homologue	NN	O	O
as	NN	O	O
a	NN	O	O
strong	NN	O	O
candidate	NN	O	O
for	NN	O	O
the	NN	O	O
AN2	NN	O	O
gene	NN	O	O
.	NN	O	O

FISH	NN	O	O
with	NN	O	O
cosmid	NN	O	O
probes	NN	O	O
has	NN	O	O
proved	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
fast	NN	O	O
and	NN	O	O
reliable	NN	O	O
technique	NN	O	O
for	NN	O	O
the	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
deletions	NN	O	O
.	NN	O	O

It	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
with	NN	O	O
limited	NN	O	O
amounts	NN	O	O
of	NN	O	O
material	NN	O	O
and	NN	O	O
has	NN	O	O
strong	NN	O	O
potential	NN	O	O
for	NN	O	O
clinical	NN	O	O
applications	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
of	NN	O	O
gene	NN	O	O
for	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
complex	NN	O	O
MHC	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Family	NN	O	O
study	NN	O	O
of	NN	O	O
a	NN	O	O
further	NN	O	O
case	NN	O	O
.	NN	O	O

Close	NN	O	O
linkage	NN	O	O
between	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
and	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
first	NN	O	O
reported	NN	O	O
by	NN	O	O
FU	NN	O	O
and	NN	O	O
co	NN	O	O
-	NN	O	O
workers	NN	O	O
in	NN	O	O
1974	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
here	NN	O	O
a	NN	O	O
pedigree	NN	O	O
of	NN	O	O
a	NN	O	O
31	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
individual	NN	O	O
with	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
Hodgkins	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
following	NN	O	O
markers	NN	O	O
were	NN	O	O
tested	NN	O	O
C2	NN	O	O
levels	NN	O	O
,	NN	O	O
factor	NN	O	O
B	NN	O	O
polymorphism	NN	O	O
,	NN	O	O
blood	NN	O	O
groups	NN	O	O
,	NN	O	O
and	NN	O	O
enzyme	NN	O	O
typing	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
and	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
both	NN	O	O
parents	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
Bf	NN	O	O
(	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
linked	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
variation	NN	O	O
of	NN	O	O
B	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
HL	NN	O	O
-	NN	O	O
A	NN	O	O
haplotypes	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
are	NN	O	O
different	NN	O	O
2	NN	O	O
,	NN	O	O
W18	NN	O	O
and	NN	O	O
W24	NN	O	O
,	NN	O	O
W18	NN	O	O
.	NN	O	O

They	NN	O	O
share	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
SD2	NN	O	O
antigen	NN	O	O
W18	NN	O	O
and	NN	O	O
the	NN	O	O
LD	NN	O	O
type	NN	O	O
7a	NN	O	O
.	NN	O	O
.	NN	O	O

Screening	NN	O	O
for	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
patients	NN	O	O
:	NN	O	O
61	NN	O	O
new	NN	O	O
patients	NN	O	O
and	NN	O	O
a	NN	O	O
summary	NN	O	O
of	NN	O	O
150	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
screening	NN	O	O
for	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
in	NN	O	O
61	NN	O	O
new	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
patients	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
summary	NN	O	O
of	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
150	NN	O	O
patients	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
a	NN	O	O
ribonuclease	NN	O	O
protection	NN	O	O
assay	NN	O	O
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
,	NN	O	O
disclosed	NN	O	O
mutations	NN	O	O
that	NN	O	O
were	NN	O	O
considered	NN	O	O
to	NN	O	O
cause	NN	O	O
significant	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
product	NN	O	O
in	NN	O	O
97	NN	O	O
of	NN	O	O
150	NN	O	O
unrelated	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
revealed	NN	O	O
the	NN	O	O
following	NN	O	O
characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
of	NN	O	O
APC	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
great	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
truncate	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
product	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
almost	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
;	NN	O	O
3	NN	O	O
)	NN	O	O
no	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
locations	NN	O	O
of	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
and	NN	O	O
extracolonic	NN	O	O
manifestations	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
;	NN	O	O
4	NN	O	O
)	NN	O	O
more	NN	O	O
than	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
base	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
were	NN	O	O
from	NN	O	O
cytosine	NN	O	O
to	NN	O	O
other	NN	O	O
nucleotides	NN	O	O
,	NN	O	O
nearly	NN	O	O
one	NN	O	O
-	NN	O	O
third	NN	O	O
of	NN	O	O
which	NN	O	O
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
GpG	NN	O	O
site	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
information	NN	O	O
helpful	NN	O	O
to	NN	O	O
an	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
will	NN	O	O
also	NN	O	O
contribute	NN	O	O
to	NN	O	O
presymptomatic	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
members	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
families	NN	O	O
.	NN	O	O
.	NN	O	O

Somatic	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
:	NN	O	O
mutation	NN	O	O
cluster	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
in	NN	O	O
63	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
(	NN	O	O
16	NN	O	O
adenomas	NN	O	B-Disease
and	NN	O	O
47	NN	O	O
carcinomas	NN	O	B-Disease
)	NN	O	O
developed	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
non	NN	O	O
-	NN	O	O
FAP	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
(	NN	O	O
LOH	NN	O	O
)	NN	O	O
at	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
locus	NN	O	O
in	NN	O	O
30	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
43	NN	O	O
other	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
one	NN	O	O
of	NN	O	O
them	NN	O	O
were	NN	O	O
point	NN	O	O
mutations	NN	O	O
;	NN	O	O
16	NN	O	O
nonsense	NN	O	O
and	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
three	NN	O	O
occurred	NN	O	O
in	NN	O	O
introns	NN	O	O
at	NN	O	O
the	NN	O	O
splicing	NN	O	O
site	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
two	NN	O	O
tumors	NN	O	B-Disease
had	NN	O	O
frameshift	NN	O	O
mutations	NN	O	O
due	NN	O	O
to	NN	O	O
deletion	NN	O	O
or	NN	O	O
insertion	NN	O	O
;	NN	O	O
nineteen	NN	O	O
of	NN	O	O
them	NN	O	O
were	NN	O	O
deletions	NN	O	O
of	NN	O	O
one	NN	O	O
to	NN	O	O
31	NN	O	O
bp	NN	O	O
and	NN	O	O
three	NN	O	O
were	NN	O	O
a	NN	O	O
1	NN	O	O
-	NN	O	O
bp	NN	O	O
insertion	NN	O	O
.	NN	O	O

One	NN	O	O
tumor	NN	O	B-Disease
had	NN	O	O
a	NN	O	O
1	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
in	NN	O	O
an	NN	O	O
intron	NN	O	O
near	NN	O	O
the	NN	O	O
splicing	NN	O	O
site	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
41	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
43	NN	O	O
mutations	NN	O	O
resulted	NN	O	O
in	NN	O	O
truncation	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
protein	NN	O	O
.	NN	O	O

Over	NN	O	O
60	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
were	NN	O	O
clustered	NN	O	O
within	NN	O	O
a	NN	O	O
small	NN	O	O
region	NN	O	O
of	NN	O	O
exon	NN	O	O
15	NN	O	O
,	NN	O	O
designated	NN	O	O
as	NN	O	O
MCR	NN	O	O
(	NN	O	O
mutation	NN	O	O
cluster	NN	O	O
region	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
accounted	NN	O	O
for	NN	O	O
less	NN	O	O
than	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
.	NN	O	O

Combining	NN	O	O
these	NN	O	O
data	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
LOH	NN	O	O
,	NN	O	O
more	NN	O	O
than	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
tumors	NN	O	B-Disease
(	NN	O	O
14	NN	O	O
adenomas	NN	O	B-Disease
and	NN	O	O
39	NN	O	O
carcinomas	NN	O	B-Disease
)	NN	O	O
had	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
more	NN	O	O
than	NN	O	O
60	NN	O	O
%	NN	O	O
(	NN	O	O
9	NN	O	O
adenomas	NN	O	B-Disease
and	NN	O	O
23	NN	O	O
carcinomas	NN	O	B-Disease
)	NN	O	O
had	NN	O	O
two	NN	O	O
mutations	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
great	NN	O	O
majority	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Constitutional	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
WT1	NN	O	O
gene	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
characterised	NN	O	O
by	NN	O	O
a	NN	O	O
typical	NN	O	O
nephropathy	NN	O	B-Disease
,	NN	O	O
genital	NN	O	B-Disease
abnormalities	NN	O	I-Disease
and	NN	O	O
also	NN	O	O
predisposes	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
patients	NN	O	O
eventually	NN	O	O
go	NN	O	O
into	NN	O	O
end	NN	O	O
stage	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
candidate	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
gene	NN	O	O
,	NN	O	O
WT1	NN	O	O
,	NN	O	O
from	NN	O	O
the	NN	O	O
11p13	NN	O	O
chromosome	NN	O	O
region	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
cloned	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
in	NN	O	O
constitutional	NN	O	O
cells	NN	O	O
from	NN	O	O
eight	NN	O	O
patients	NN	O	O
and	NN	O	O
have	NN	O	O
shown	NN	O	O
heterozygous	NN	O	O
mutations	NN	O	O
in	NN	O	O
six	NN	O	O
of	NN	O	O
them	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
in	NN	O	O
exon	NN	O	O
9	NN	O	O
,	NN	O	O
all	NN	O	O
resulting	NN	O	O
in	NN	O	O
missense	NN	O	O
mutations	NN	O	O
.	NN	O	O

Three	NN	O	O
were	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
1180	NN	O	O
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
arg	NN	O	O
>	NN	O	O
trp	NN	O	O
amino	NN	O	O
acid	NN	O	O
change	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
was	NN	O	O
at	NN	O	O
position	NN	O	O
1186	NN	O	O
converting	NN	O	O
an	NN	O	O
asp	NN	O	O
>	NN	O	O
asn	NN	O	O
in	NN	O	O
the	NN	O	O
predicted	NN	O	O
resultant	NN	O	O
protein	NN	O	O
.	NN	O	O

One	NN	O	O
patient	NN	O	O
had	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
,	NN	O	O
converting	NN	O	O
an	NN	O	O
arg	NN	O	O
>	NN	O	O
his	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
base	NN	O	O
pair	NN	O	O
insertion	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
821	NN	O	O
in	NN	O	O
exon	NN	O	O
6	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
patient	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
find	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
patient	NN	O	O
despite	NN	O	O
complete	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
genomic	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
last	NN	O	O
patient	NN	O	O
carried	NN	O	O
a	NN	O	O
constitutional	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
11p13	NN	O	O
region	NN	O	O
and	NN	O	O
no	NN	O	O
additional	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
obvious	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
and	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
WT1	NN	O	O
gene	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
kidney	NN	O	O
and	NN	O	O
the	NN	O	O
genito	NN	O	O
-	NN	O	O
urinary	NN	O	O
system	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
disequilibrium	NN	O	O
mapping	NN	O	O
in	NN	O	O
isolated	NN	O	O
founder	NN	O	O
populations	NN	O	O
:	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
in	NN	O	O
Finland	NN	O	O
.	NN	O	O

Linkage	NN	O	O
disequilibrium	NN	O	O
mapping	NN	O	O
in	NN	O	O
isolated	NN	O	O
populations	NN	O	O
provides	NN	O	O
a	NN	O	O
powerful	NN	O	O
tool	NN	O	O
for	NN	O	O
fine	NN	O	O
structure	NN	O	O
localization	NN	O	O
of	NN	O	O
disease	NN	O	O
genes	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
Luria	NN	O	O
and	NN	O	O
Delbrucks	NN	O	O
classical	NN	O	O
methods	NN	O	O
for	NN	O	O
analysing	NN	O	O
bacterial	NN	O	O
cultures	NN	O	O
are	NN	O	O
adapted	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
human	NN	O	O
isolated	NN	O	O
founder	NN	O	O
populations	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
estimate	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
the	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
between	NN	O	O
a	NN	O	O
disease	NN	O	O
locus	NN	O	O
and	NN	O	O
a	NN	O	O
marker	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
the	NN	O	O
expected	NN	O	O
degree	NN	O	O
of	NN	O	O
allelic	NN	O	O
homogeneity	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
the	NN	O	O
mutation	NN	O	O
rate	NN	O	O
of	NN	O	O
marker	NN	O	O
loci	NN	O	O
.	NN	O	O

Using	NN	O	O
these	NN	O	O
methods	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
striking	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
for	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
(	NN	O	O
DTD	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
Finland	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
DTD	NN	O	B-Disease
gene	NN	O	O
should	NN	O	O
lie	NN	O	O
within	NN	O	O
0	NN	O	O
.	NN	O	O

06	NN	O	O
centimorgans	NN	O	O
(	NN	O	O
or	NN	O	O
about	NN	O	O
60	NN	O	O
kilobases	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
CSF1R	NN	O	O
gene	NN	O	O
.	NN	O	O

Predictions	NN	O	O
about	NN	O	O
allelic	NN	O	O
homogeneity	NN	O	O
in	NN	O	O
Finland	NN	O	O
and	NN	O	O
mutation	NN	O	O
rates	NN	O	O
in	NN	O	O
simple	NN	O	O
sequence	NN	O	O
repeats	NN	O	O
are	NN	O	O
confirmed	NN	O	O
by	NN	O	O
independent	NN	O	O
observations	NN	O	O
.	NN	O	O

The	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
refinement	NN	O	O
of	NN	O	O
the	NN	O	O
localization	NN	O	O
on	NN	O	O
Xp	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
another	NN	O	O
closely	NN	O	O
linked	NN	O	O
marker	NN	O	O
locus	NN	O	O
,	NN	O	O
OATL1	NN	O	O
.	NN	O	O

The	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
proximal	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
between	NN	O	O
the	NN	O	O
DXS14	NN	O	O
and	NN	O	O
DXS7	NN	O	O
loci	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
further	NN	O	O
segregation	NN	O	O
analysis	NN	O	O
has	NN	O	O
been	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
WAS	NN	O	B-Disease
family	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
additional	NN	O	O
marker	NN	O	O
probe	NN	O	O
,	NN	O	O
HOATL1	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
and	NN	O	O
OATL1	NN	O	O
loci	NN	O	O
(	NN	O	O
Z	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
08	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
00	NN	O	O
)	NN	O	O
and	NN	O	O
localize	NN	O	O
the	NN	O	O
TIMP	NN	O	O
,	NN	O	O
OATL1	NN	O	O
,	NN	O	O
DXS255	NN	O	O
,	NN	O	O
and	NN	O	O
WAS	NN	O	B-Disease
loci	NN	O	O
distal	NN	O	O
to	NN	O	O
DXS146	NN	O	O
and	NN	O	O
the	NN	O	O
OATL1	NN	O	O
and	NN	O	O
WAS	NN	O	B-Disease
loci	NN	O	O
proximal	NN	O	O
to	NN	O	O
TIMP	NN	O	O
.	NN	O	O

These	NN	O	O
linkage	NN	O	O
data	NN	O	O
narrow	NN	O	O
the	NN	O	O
boundaries	NN	O	O
within	NN	O	O
which	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
locus	NN	O	O
maps	NN	O	O
to	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
region	NN	O	O
bracketed	NN	O	O
by	NN	O	O
TIMP	NN	O	O
and	NN	O	O
DXS146	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
loci	NN	O	O
order	NN	O	O
Xpter	NN	O	O
-	NN	O	O
DXS7	NN	O	O
-	NN	O	O
TIMP	NN	O	O
-	NN	O	O
(	NN	O	O
OATL1	NN	O	O
,	NN	O	O
WAS	NN	O	O
,	NN	O	O
DXS255	NN	O	O
)	NN	O	O
-	NN	O	O
DXS146	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
an	NN	O	O
unstable	NN	O	O
fragment	NN	O	O
of	NN	O	O
DNA	NN	O	O
specific	NN	O	O
to	NN	O	O
individuals	NN	O	O
with	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
adult	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
with	NN	O	O
a	NN	O	O
prevalence	NN	O	O
of	NN	O	O
2	NN	O	O
-	NN	O	O
14	NN	O	O
per	NN	O	O
100	NN	O	O
,	NN	O	O
000	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
disease	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
progressive	NN	O	O
muscle	NN	O	B-Disease
weakness	NN	O	I-Disease
and	NN	O	O
sustained	NN	O	B-Disease
muscle	NN	O	I-Disease
contraction	NN	O	I-Disease
,	NN	O	O
often	NN	O	O
with	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
accompanying	NN	O	O
symptoms	NN	O	O
.	NN	O	O

The	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
and	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
show	NN	O	O
extreme	NN	O	O
variation	NN	O	O
,	NN	O	O
both	NN	O	O
within	NN	O	O
and	NN	O	O
between	NN	O	O
families	NN	O	O
.	NN	O	O

Despite	NN	O	O
its	NN	O	O
clinical	NN	O	O
variability	NN	O	O
,	NN	O	O
this	NN	O	O
dominant	NN	O	O
condition	NN	O	O
segregates	NN	O	O
as	NN	O	O
a	NN	O	O
single	NN	O	O
locus	NN	O	O
at	NN	O	O
chromosome	NN	O	O
19q13	NN	O	O
.	NN	O	O

3	NN	O	O
in	NN	O	O
every	NN	O	O
population	NN	O	O
studied	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
flanked	NN	O	O
by	NN	O	O
the	NN	O	O
tightly	NN	O	O
linked	NN	O	O
genetic	NN	O	O
markers	NN	O	O
ERCC1	NN	O	O
proximally	NN	O	O
and	NN	O	O
D19S51	NN	O	O
distally	NN	O	O
;	NN	O	O
these	NN	O	O
define	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
critical	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
an	NN	O	O
expressed	NN	O	O
sequence	NN	O	O
from	NN	O	O
this	NN	O	O
region	NN	O	O
which	NN	O	O
detects	NN	O	O
a	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
that	NN	O	O
is	NN	O	O
larger	NN	O	O
in	NN	O	O
affected	NN	O	O
individuals	NN	O	O
than	NN	O	O
in	NN	O	O
normal	NN	O	O
siblings	NN	O	O
or	NN	O	O
unaffected	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
size	NN	O	O
of	NN	O	O
this	NN	O	O
fragment	NN	O	O
varies	NN	O	O
between	NN	O	O
affected	NN	O	O
siblings	NN	O	O
,	NN	O	O
and	NN	O	O
increases	NN	O	O
in	NN	O	O
size	NN	O	O
through	NN	O	O
generations	NN	O	O
in	NN	O	O
parallel	NN	O	O
with	NN	O	O
increasing	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
postulate	NN	O	O
that	NN	O	O
this	NN	O	O
unstable	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
is	NN	O	O
the	NN	O	O
molecular	NN	O	O
feature	NN	O	O
that	NN	O	O
underlies	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

Common	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
at	NN	O	O
the	NN	O	O
rearrangement	NN	O	O
sites	NN	O	O
of	NN	O	O
a	NN	O	O
constitutional	NN	O	O
X	NN	O	O
/	NN	O	O
autosome	NN	O	O
translocation	NN	O	O
and	NN	O	O
associated	NN	O	O
deletion	NN	O	O
.	NN	O	O

Reciprocal	NN	O	O
chromosome	NN	O	O
translocations	NN	O	O
are	NN	O	O
common	NN	O	O
de	NN	O	O
novo	NN	O	O
rearrangements	NN	O	O
that	NN	O	O
occur	NN	O	O
randomly	NN	O	O
throughout	NN	O	O
the	NN	O	O
human	NN	O	O
genome	NN	O	O
.	NN	O	O

To	NN	O	O
learn	NN	O	O
about	NN	O	O
causative	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
a	NN	O	O
cytologically	NN	O	O
balanced	NN	O	O
constitutional	NN	O	O
reciprocal	NN	O	O
translocation	NN	O	O
,	NN	O	O
t	NN	O	O
(	NN	O	O
X	NN	O	O
;	NN	O	O
4	NN	O	O
)	NN	O	O
(	NN	O	O
p21	NN	O	O
.	NN	O	O
2	NN	O	O
;	NN	O	O
q31	NN	O	O
.	NN	O	O
22	NN	O	O
)	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
girl	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Physical	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
derivative	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
after	NN	O	O
their	NN	O	O
separation	NN	O	O
in	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
,	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
translocation	NN	O	O
disrupts	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
Xp21	NN	O	O
within	NN	O	O
the	NN	O	O
18	NN	O	O
-	NN	O	O
kb	NN	O	O
intron	NN	O	O
16	NN	O	O
.	NN	O	O

Restriction	NN	O	O
mapping	NN	O	O
and	NN	O	O
sequencing	NN	O	O
of	NN	O	O
clones	NN	O	O
that	NN	O	O
span	NN	O	O
both	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
corresponding	NN	O	O
normal	NN	O	O
regions	NN	O	O
indicate	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
approximately	NN	O	O
5	NN	O	O
kb	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
derivative	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
with	NN	O	O
4	NN	O	O
-	NN	O	O
6	NN	O	O
bp	NN	O	O
deleted	NN	O	O
from	NN	O	O
chromosome	NN	O	O
4	NN	O	O
.	NN	O	O

RFLP	NN	O	O
and	NN	O	O
Southern	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
de	NN	O	O
novo	NN	O	O
translocation	NN	O	O
is	NN	O	O
a	NN	O	O
paternal	NN	O	O
origin	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
fathers	NN	O	O
X	NN	O	O
chromosome	NN	O	O
contains	NN	O	O
the	NN	O	O
DNA	NN	O	O
that	NN	O	O
is	NN	O	O
deleted	NN	O	O
in	NN	O	O
the	NN	O	O
derivative	NN	O	O
X	NN	O	O
.	NN	O	O

Most	NN	O	O
likely	NN	O	O
,	NN	O	O
deletion	NN	O	O
and	NN	O	O
translation	NN	O	O
arose	NN	O	O
simultaneously	NN	O	O
from	NN	O	O
a	NN	O	O
complex	NN	O	O
rearrangement	NN	O	O
event	NN	O	O
that	NN	O	O
involves	NN	O	O
three	NN	O	O
chromosomal	NN	O	O
breakpoints	NN	O	O
.	NN	O	O

Short	NN	O	O
regions	NN	O	O
of	NN	O	O
sequence	NN	O	O
homology	NN	O	O
were	NN	O	O
present	NN	O	O
at	NN	O	O
the	NN	O	O
three	NN	O	O
sites	NN	O	O
.	NN	O	O

A	NN	O	O
5	NN	O	O
-	NN	O	O
bp	NN	O	O
sequence	NN	O	O
,	NN	O	O
GGAAT	NN	O	O
,	NN	O	O
found	NN	O	O
exactly	NN	O	O
at	NN	O	O
the	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
on	NN	O	O
both	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
X	NN	O	O
and	NN	O	O
4	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
preserved	NN	O	O
only	NN	O	O
on	NN	O	O
the	NN	O	O
der	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
chromosome	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
derived	NN	O	O
sequence	NN	O	O
GGAATCA	NN	O	O
has	NN	O	O
been	NN	O	O
lost	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
der	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
chromosome	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
matches	NN	O	O
an	NN	O	O
inverted	NN	O	O
GAATCA	NN	O	O
sequence	NN	O	O
present	NN	O	O
on	NN	O	O
the	NN	O	O
opposite	NN	O	O
strand	NN	O	O
exactly	NN	O	O
at	NN	O	O
the	NN	O	O
other	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
deleted	NN	O	O
5	NN	O	O
-	NN	O	O
kb	NN	O	O
fragment	NN	O	O
.	NN	O	O

A	NN	O	O
genetic	NN	O	O
etiology	NN	O	O
for	NN	O	O
DiGeorge	NN	O	B-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
consistent	NN	O	O
deletions	NN	O	O
and	NN	O	O
microdeletions	NN	O	O
of	NN	O	O
22q11	NN	O	O
.	NN	O	O

DiGeorge	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
DGS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
developmental	NN	O	O
field	NN	O	O
defect	NN	O	O
of	NN	O	O
the	NN	O	O
third	NN	O	O
and	NN	O	O
fourth	NN	O	O
pharyngeal	NN	O	O
pouches	NN	O	O
,	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
aplasia	NN	O	B-Disease
or	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
thymus	NN	O	I-Disease
and	NN	O	I-Disease
parathyroid	NN	O	I-Disease
glands	NN	O	I-Disease
and	NN	O	O
by	NN	O	O
conotruncal	NN	O	B-Disease
cardiac	NN	O	I-Disease
malformations	NN	O	I-Disease
.	NN	O	O

Cytogenetic	NN	O	O
studies	NN	O	O
support	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
DGS	NN	O	B-Disease
critical	NN	O	O
region	NN	O	O
in	NN	O	O
band	NN	O	O
22q11	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
clinical	NN	O	O
,	NN	O	O
cytogenetic	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
studies	NN	O	O
of	NN	O	O
14	NN	O	O
patients	NN	O	O
with	NN	O	O
DGS	NN	O	B-Disease
.	NN	O	O

Chromosome	NN	O	O
analysis	NN	O	O
,	NN	O	O
utilizing	NN	O	O
high	NN	O	O
-	NN	O	O
resolution	NN	O	O
banding	NN	O	O
techniques	NN	O	O
,	NN	O	O
detected	NN	O	O
interstitial	NN	O	O
deletions	NN	O	O
in	NN	O	O
five	NN	O	O
probands	NN	O	O
and	NN	O	O
was	NN	O	O
inconclusive	NN	O	O
for	NN	O	O
a	NN	O	O
deletion	NN	O	O
in	NN	O	O
three	NN	O	O
probands	NN	O	O
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
six	NN	O	O
patients	NN	O	O
had	NN	O	O
normal	NN	O	O
karyotypes	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
detected	NN	O	O
DNA	NN	O	O
deletions	NN	O	O
in	NN	O	O
all	NN	O	O
14	NN	O	O
probands	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
10	NN	O	O
loci	NN	O	O
tested	NN	O	O
,	NN	O	O
D22S75	NN	O	O
and	NN	O	O
D22S259	NN	O	O
,	NN	O	O
are	NN	O	O
deleted	NN	O	O
in	NN	O	O
all	NN	O	O
14	NN	O	O
patients	NN	O	O
.	NN	O	O

A	NN	O	O
third	NN	O	O
locus	NN	O	O
,	NN	O	O
D22S66	NN	O	O
,	NN	O	O
is	NN	O	O
deleted	NN	O	O
in	NN	O	O
the	NN	O	O
eight	NN	O	O
DGS	NN	O	B-Disease
probands	NN	O	O
tested	NN	O	O
.	NN	O	O

Physical	NN	O	O
mapping	NN	O	O
using	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
places	NN	O	O
D22S66	NN	O	O
between	NN	O	O
D22S75	NN	O	O
and	NN	O	O
D22S259	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
should	NN	O	O
be	NN	O	O
deleted	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
six	NN	O	O
cases	NN	O	O
.	NN	O	O

Parent	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
origin	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
in	NN	O	O
five	NN	O	O
families	NN	O	O
.	NN	O	O

Four	NN	O	O
probands	NN	O	O
failed	NN	O	O
to	NN	O	O
inherit	NN	O	O
a	NN	O	O
maternal	NN	O	O
allele	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
failed	NN	O	O
to	NN	O	O
inherit	NN	O	O
a	NN	O	O
paternal	NN	O	O
allele	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
these	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
of	NN	O	O
six	NN	O	O
maternally	NN	O	O
and	NN	O	O
five	NN	O	O
paternally	NN	O	O
derived	NN	O	O
unbalanced	NN	O	O
-	NN	O	O
translocation	NN	O	O
DGS	NN	O	B-Disease
probands	NN	O	O
in	NN	O	O
the	NN	O	O
literature	NN	O	O
,	NN	O	O
parent	NN	O	O
of	NN	O	O
origin	NN	O	O
or	NN	O	O
imprinting	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
DGS	NN	O	B-Disease
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
three	NN	O	O
loci	NN	O	O
in	NN	O	O
all	NN	O	O
14	NN	O	O
DGS	NN	O	B-Disease
probands	NN	O	O
begins	NN	O	O
to	NN	O	O
delineate	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22	NN	O	O
critical	NN	O	O
for	NN	O	O
DGS	NN	O	B-Disease
and	NN	O	O
confirms	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
submicroscopic	NN	O	O
deletions	NN	O	O
of	NN	O	O
22q11	NN	O	O
are	NN	O	O
etiologic	NN	O	O
in	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
cases	NN	O	O
.	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
normal	NN	O	O
allele	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
an	NN	O	O
adrenocortical	NN	O	B-Disease
carcinoma	NN	O	I-Disease
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
.	NN	O	O

Endocrine	NN	O	B-Disease
neoplasms	NN	O	I-Disease
have	NN	O	O
been	NN	O	O
reported	NN	O	O
occasionally	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

An	NN	O	O
adrenocorotical	NN	O	B-Disease
carcinoma	NN	O	I-Disease
was	NN	O	O
studied	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
FAP	NN	O	B-Disease
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
(	NN	O	O
LOH	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
this	NN	O	O
carcinoma	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
evidence	NN	O	O
was	NN	O	O
obtained	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
allele	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
of	NN	O	O
LOH	NN	O	O
at	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
locus	NN	O	O
in	NN	O	O
adrenocortical	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
present	NN	O	O
results	NN	O	O
and	NN	O	O
our	NN	O	O
previous	NN	O	O
data	NN	O	O
on	NN	O	O
LOH	NN	O	O
in	NN	O	O
a	NN	O	O
recurring	NN	O	O
desmoid	NN	O	B-Disease
tumor	NN	O	I-Disease
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
mutant	NN	O	O
/	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
condition	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
may	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
benign	NN	O	B-Disease
tumors	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
that	NN	O	O
functional	NN	O	O
loss	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
leads	NN	O	O
to	NN	O	O
development	NN	O	O
of	NN	O	O
tumors	NN	O	B-Disease
not	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
colon	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
other	NN	O	O
various	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
body	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Oncogenic	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
exon	NN	O	O
20	NN	O	O
of	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
in	NN	O	O
families	NN	O	O
showing	NN	O	O
incomplete	NN	O	O
penetrance	NN	O	O
and	NN	O	O
mild	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
retinoblastoma	NN	O	B-Disease
-	NN	O	O
predisposition	NN	O	O
gene	NN	O	O
,	NN	O	O
RB1	NN	O	O
,	NN	O	O
segregates	NN	O	O
as	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
trait	NN	O	O
with	NN	O	O
high	NN	O	O
(	NN	O	O
90	NN	O	O
%	NN	O	O
)	NN	O	O
penetrance	NN	O	O
.	NN	O	O

Certain	NN	O	O
families	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
show	NN	O	O
an	NN	O	O
unusual	NN	O	O
low	NN	O	O
-	NN	O	O
penetrance	NN	O	O
phenotype	NN	O	O
with	NN	O	O
many	NN	O	O
individuals	NN	O	O
being	NN	O	O
unaffected	NN	O	O
,	NN	O	O
unilaterally	NN	O	O
affected	NN	O	O
,	NN	O	O
or	NN	O	O
with	NN	O	O
evidence	NN	O	O
of	NN	O	O
spontaneously	NN	O	O
regressed	NN	O	O
tumors	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
and	NN	O	O
PCR	NN	O	O
sequencing	NN	O	O
to	NN	O	O
study	NN	O	O
two	NN	O	O
such	NN	O	O
families	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
exon	NN	O	O
20	NN	O	O
of	NN	O	O
RB1	NN	O	O
in	NN	O	O
both	NN	O	O
cases	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
family	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
codon	NN	O	O
661	NN	O	O
converts	NN	O	O
an	NN	O	O
arginine	NN	O	O
(	NN	O	O
CGG	NN	O	O
)	NN	O	O
to	NN	O	O
a	NN	O	O
tryptophan	NN	O	O
(	NN	O	O
TGG	NN	O	O
)	NN	O	O
codon	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
incomplete	NN	O	O
penetrance	NN	O	O
and	NN	O	O
mild	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
virtually	NN	O	O
all	NN	O	O
patients	NN	O	O
,	NN	O	O
possibly	NN	O	O
indicating	NN	O	O
that	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
changes	NN	O	O
may	NN	O	O
modify	NN	O	O
protein	NN	O	O
structure	NN	O	O
/	NN	O	O
function	NN	O	O
such	NN	O	O
that	NN	O	O
tumorigenesis	NN	O	O
is	NN	O	O
not	NN	O	O
inevitable	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
second	NN	O	O
family	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
codon	NN	O	O
675	NN	O	O
is	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
transversion	NN	O	O
that	NN	O	O
converts	NN	O	O
a	NN	O	O
glutamine	NN	O	O
(	NN	O	O
GAA	NN	O	O
)	NN	O	O
to	NN	O	O
a	NN	O	O
stop	NN	O	O
(	NN	O	O
TAA	NN	O	O
)	NN	O	O
codon	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
mutation	NN	O	O
also	NN	O	O
occurs	NN	O	O
near	NN	O	O
a	NN	O	O
potential	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
acceptor	NN	O	O
site	NN	O	O
,	NN	O	O
raising	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
less	NN	O	O
severely	NN	O	O
disrupted	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Late	NN	O	O
-	NN	O	O
onset	NN	O	O
metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
:	NN	O	O
molecular	NN	O	O
pathology	NN	O	O
in	NN	O	O
two	NN	O	O
siblings	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
new	NN	O	O
allele	NN	O	O
at	NN	O	O
the	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
locus	NN	O	O
causing	NN	O	O
late	NN	O	O
-	NN	O	O
onset	NN	O	O
metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
that	NN	O	O
allele	NN	O	O
arginine84	NN	O	O
,	NN	O	O
a	NN	O	O
residue	NN	O	O
that	NN	O	O
is	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
arylsulfatase	NN	O	O
gene	NN	O	O
family	NN	O	O
,	NN	O	O
is	NN	O	O
replaced	NN	O	O
by	NN	O	O
glutamine	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
alleles	NN	O	O
that	NN	O	O
cause	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
MLD	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
arginine84	NN	O	O
to	NN	O	O
glutamine	NN	O	O
substitution	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
some	NN	O	O
residual	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
genotypes	NN	O	O
,	NN	O	O
ARSA	NN	O	O
activities	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
data	NN	O	O
on	NN	O	O
4	NN	O	O
individuals	NN	O	O
carrying	NN	O	O
the	NN	O	O
allele	NN	O	O
of	NN	O	O
81	NN	O	O
patients	NN	O	O
with	NN	O	O
MLD	NN	O	B-Disease
examined	NN	O	O
,	NN	O	O
further	NN	O	O
validates	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
different	NN	O	O
degrees	NN	O	O
of	NN	O	O
residual	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
are	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
phenotypical	NN	O	O
variation	NN	O	O
in	NN	O	O
MLD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Trisomy	NN	O	B-Disease
15	NN	O	I-Disease
with	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
15	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
maternal	NN	O	B-Disease
disomy	NN	O	I-Disease
.	NN	O	O

Uniparental	NN	O	B-Disease
disomy	NN	O	I-Disease
has	NN	O	O
recently	NN	O	O
been	NN	O	O
recognized	NN	O	O
to	NN	O	O
cause	NN	O	O
human	NN	O	O
disorders	NN	O	O
,	NN	O	O
including	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
particularly	NN	O	O
instructive	NN	O	O
case	NN	O	O
which	NN	O	O
raises	NN	O	O
important	NN	O	O
issues	NN	O	O
concerning	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
producing	NN	O	O
uniparental	NN	O	B-Disease
disomy	NN	O	I-Disease
and	NN	O	O
whose	NN	O	O
evaluation	NN	O	O
provides	NN	O	O
evidence	NN	O	O
that	NN	O	O
trisomy	NN	O	O
may	NN	O	O
precede	NN	O	O
uniparental	NN	O	B-Disease
disomy	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
fetus	NN	O	O
.	NN	O	O

Chorionic	NN	O	O
villus	NN	O	O
sampling	NN	O	O
performed	NN	O	O
for	NN	O	O
advanced	NN	O	O
maternal	NN	O	O
age	NN	O	O
revealed	NN	O	O
trisomy	NN	O	B-Disease
15	NN	O	I-Disease
in	NN	O	O
all	NN	O	O
direct	NN	O	O
and	NN	O	O
cultured	NN	O	O
cells	NN	O	O
,	NN	O	O
though	NN	O	O
the	NN	O	O
fetus	NN	O	O
appeared	NN	O	O
normal	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
analysis	NN	O	O
of	NN	O	O
amniocytes	NN	O	O
obtained	NN	O	O
at	NN	O	O
15	NN	O	O
wk	NN	O	O
was	NN	O	O
normal	NN	O	O
in	NN	O	O
over	NN	O	O
100	NN	O	O
cells	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
child	NN	O	O
was	NN	O	O
hypotonic	NN	O	B-Disease
at	NN	O	O
birth	NN	O	O
,	NN	O	O
and	NN	O	O
high	NN	O	O
-	NN	O	O
resolution	NN	O	O
banding	NN	O	O
failed	NN	O	O
to	NN	O	O
reveal	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
,	NN	O	O
a	NN	O	O
deletion	NN	O	O
which	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
50	NN	O	O
%	NN	O	O
-	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Over	NN	O	O
time	NN	O	O
,	NN	O	O
typical	NN	O	O
features	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
developed	NN	O	O
.	NN	O	O

Molecular	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
using	NN	O	O
probes	NN	O	O
for	NN	O	O
chromosome	NN	O	O
15	NN	O	O
revealed	NN	O	O
maternal	NN	O	O
disomy	NN	O	O
.	NN	O	O

Maternal	NN	O	O
nondisjunction	NN	O	O
with	NN	O	O
fertilization	NN	O	O
of	NN	O	O
a	NN	O	O
disomic	NN	O	O
egg	NN	O	O
by	NN	O	O
a	NN	O	O
normal	NN	O	O
sperm	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
15	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
likely	NN	O	O
cause	NN	O	O
of	NN	O	O
confined	NN	O	O
placental	NN	O	O
mosaicism	NN	O	O
and	NN	O	O
uniparental	NN	O	B-Disease
disomy	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
case	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
advanced	NN	O	O
maternal	NN	O	O
age	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
predisposing	NN	O	O
factor	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
amelogenesis	NN	O	I-Disease
imperfecta	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
AMELX	NN	O	O
gene	NN	O	O
located	NN	O	O
at	NN	O	O
Xp22	NN	O	O
.	NN	O	O

1	NN	O	O
-	NN	O	O
p22	NN	O	O
.	NN	O	O

3	NN	O	O
encodes	NN	O	O
for	NN	O	O
the	NN	O	O
enamel	NN	O	O
protein	NN	O	O
amelogenin	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
as	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
dental	NN	O	I-Disease
abnormality	NN	O	I-Disease
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
amelogenesis	NN	O	I-Disease
imperfecta	NN	O	I-Disease
(	NN	O	O
XAI	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Three	NN	O	O
families	NN	O	O
with	NN	O	O
XAI	NN	O	B-Disease
have	NN	O	O
been	NN	O	O
investigated	NN	O	O
using	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
markers	NN	O	O
flanking	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
AMELX	NN	O	O
.	NN	O	O

Using	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
,	NN	O	O
linkage	NN	O	O
was	NN	O	O
established	NN	O	O
between	NN	O	O
XAI	NN	O	B-Disease
and	NN	O	O
several	NN	O	O
of	NN	O	O
these	NN	O	O
markers	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
combined	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
6	NN	O	O
.	NN	O	O

05	NN	O	O
for	NN	O	O
DXS16	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

04	NN	O	O
04	NN	O	O
.	NN	O	O

This	NN	O	O
supports	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
AMELX	NN	O	O
,	NN	O	O
located	NN	O	O
close	NN	O	O
to	NN	O	O
DXS16	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
XAI	NN	O	B-Disease
disease	NN	O	I-Disease
process	NN	O	O
(	NN	O	O
AIH1	NN	O	O
)	NN	O	O
in	NN	O	O
those	NN	O	O
families	NN	O	O
.	NN	O	O

Using	NN	O	O
multipoint	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
,	NN	O	O
the	NN	O	O
combined	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
for	NN	O	O
these	NN	O	O
two	NN	O	O
families	NN	O	O
was	NN	O	O
7	NN	O	O
.	NN	O	O

30	NN	O	O
for	NN	O	O
a	NN	O	O
location	NN	O	O
of	NN	O	O
AIH1	NN	O	O
at	NN	O	O
2	NN	O	O
cM	NN	O	O
distal	NN	O	O
to	NN	O	O
DXS16	NN	O	O
.	NN	O	O

The	NN	O	O
support	NN	O	O
interval	NN	O	O
around	NN	O	O
this	NN	O	O
location	NN	O	O
extended	NN	O	O
about	NN	O	O
8	NN	O	O
cM	NN	O	O
proximal	NN	O	O
to	NN	O	O
DXS92	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
AIH1	NN	O	O
location	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
precisely	NN	O	O
defined	NN	O	O
by	NN	O	O
multipoint	NN	O	O
mapping	NN	O	O
.	NN	O	O

Study	NN	O	O
of	NN	O	O
recombination	NN	O	O
events	NN	O	O
indicated	NN	O	O
that	NN	O	O
AIH1	NN	O	O
lies	NN	O	O
in	NN	O	O
the	NN	O	O
interval	NN	O	O
between	NN	O	O
DXS143	NN	O	O
and	NN	O	O
DXS85	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
significant	NN	O	O
evidence	NN	O	O
against	NN	O	O
linkage	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	O
family	NN	O	O
,	NN	O	O
indicating	NN	O	O
locus	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
XAI	NN	O	B-Disease
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
with	NN	O	O
markers	NN	O	O
on	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
showed	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
DXS144E	NN	O	O
and	NN	O	O
F9	NN	O	O
with	NN	O	O
no	NN	O	O
recombination	NN	O	O
with	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
markers	NN	O	O
.	NN	O	O

Two	NN	O	O
-	NN	O	O
point	NN	O	O
analysis	NN	O	O
gave	NN	O	O
a	NN	O	O
peak	NN	O	O
lod	NN	O	O
score	NN	O	O
at	NN	O	O
DXS144E	NN	O	O
with	NN	O	O
a	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

83	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
peak	NN	O	O
lod	NN	O	O
score	NN	O	O
in	NN	O	O
multipoint	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

84	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

The	NN	O	O
support	NN	O	O
interval	NN	O	O
extended	NN	O	O
9	NN	O	O
cM	NN	O	O
proximal	NN	O	O
to	NN	O	O
DXS144E	NN	O	O
and	NN	O	O
14	NN	O	O
cM	NN	O	O
distal	NN	O	O
to	NN	O	O
F9	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O
.	NN	O	O

Multiple	NN	O	O
origins	NN	O	O
for	NN	O	O
phenylketonuria	NN	O	B-Disease
in	NN	O	O
Europe	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
disorder	NN	O	B-Disease
of	NN	O	I-Disease
amino	NN	O	I-Disease
acid	NN	O	I-Disease
metabolism	NN	O	I-Disease
prevalent	NN	O	O
among	NN	O	O
Caucasians	NN	O	O
and	NN	O	O
other	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
,	NN	O	O
is	NN	O	O
caused	NN	O	O
primarily	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
hepatic	NN	O	I-Disease
enzyme	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

PKU	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
highly	NN	O	O
heterogeneous	NN	O	O
disorder	NN	O	O
,	NN	O	O
with	NN	O	O
more	NN	O	O
than	NN	O	O
60	NN	O	O
molecular	NN	O	O
lesions	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
haplotype	NN	O	O
associations	NN	O	O
,	NN	O	O
relative	NN	O	O
frequencies	NN	O	O
,	NN	O	O
and	NN	O	O
distributions	NN	O	O
of	NN	O	O
five	NN	O	O
prevalent	NN	O	O
PAH	NN	O	O
mutations	NN	O	O
(	NN	O	O
R158Q	NN	O	O
,	NN	O	O
R261Q	NN	O	O
,	NN	O	O
IVS10nt546	NN	O	O
,	NN	O	O
R408W	NN	O	O
,	NN	O	O
and	NN	O	O
IVS12n1	NN	O	O
)	NN	O	O
were	NN	O	O
established	NN	O	O
in	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
European	NN	O	O
sample	NN	O	O
population	NN	O	O
and	NN	O	O
subsequently	NN	O	O
were	NN	O	O
examined	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
potential	NN	O	O
roles	NN	O	O
of	NN	O	O
several	NN	O	O
genetic	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
explaining	NN	O	O
the	NN	O	O
present	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
PKU	NN	O	B-Disease
alleles	NN	O	O
.	NN	O	O

Each	NN	O	O
of	NN	O	O
these	NN	O	O
five	NN	O	O
mutations	NN	O	O
was	NN	O	O
strongly	NN	O	O
associated	NN	O	O
with	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
more	NN	O	O
than	NN	O	O
70	NN	O	O
chromosomal	NN	O	O
haplotypes	NN	O	O
defined	NN	O	O
by	NN	O	O
eight	NN	O	O
RFLPs	NN	O	O
in	NN	O	O
or	NN	O	O
near	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
arose	NN	O	O
through	NN	O	O
a	NN	O	O
single	NN	O	O
founding	NN	O	O
event	NN	O	O
that	NN	O	O
occurred	NN	O	O
within	NN	O	O
time	NN	O	O
periods	NN	O	O
ranging	NN	O	O
from	NN	O	O
several	NN	O	O
hundred	NN	O	O
to	NN	O	O
several	NN	O	O
thousand	NN	O	O
years	NN	O	O
ago	NN	O	O
.	NN	O	O

From	NN	O	O
the	NN	O	O
significant	NN	O	O
differences	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
relative	NN	O	O
frequencies	NN	O	O
and	NN	O	O
distributions	NN	O	O
of	NN	O	O
these	NN	O	O
five	NN	O	O
alleles	NN	O	O
throughout	NN	O	O
Europe	NN	O	O
,	NN	O	O
four	NN	O	O
of	NN	O	O
these	NN	O	O
putative	NN	O	O
founding	NN	O	O
events	NN	O	O
could	NN	O	O
be	NN	O	O
localized	NN	O	O
to	NN	O	O
specific	NN	O	O
ethnic	NN	O	O
subgroups	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
there	NN	O	O
were	NN	O	O
multiple	NN	O	O
,	NN	O	O
geographically	NN	O	O
and	NN	O	O
ethnically	NN	O	O
distinct	NN	O	O
origins	NN	O	O
for	NN	O	O
PKU	NN	O	B-Disease
within	NN	O	O
the	NN	O	O
European	NN	O	O
population	NN	O	O
.	NN	O	O
.	NN	O	O

Complement	NN	O	B-Disease
factor	NN	O	I-Disease
2	NN	O	I-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
clinical	NN	O	O
and	NN	O	O
serological	NN	O	O
family	NN	O	O
study	NN	O	O
.	NN	O	O

Inherited	NN	O	B-Disease
complement	NN	O	I-Disease
deficiencies	NN	O	I-Disease
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
connective	NN	O	O
tissue	NN	O	O
diseases	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
with	NN	O	O
inherited	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
factor	NN	O	I-Disease
2	NN	O	I-Disease
(	NN	O	O
C2	NN	O	O
)	NN	O	O
is	NN	O	O
described	NN	O	O
in	NN	O	O
which	NN	O	O
two	NN	O	O
family	NN	O	O
members	NN	O	O
with	NN	O	O
homozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
developed	NN	O	O
cutaneous	NN	O	B-Disease
vasculitis	NN	O	I-Disease
and	NN	O	O
sicca	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
other	NN	O	O
family	NN	O	O
members	NN	O	O
had	NN	O	O
heterozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
each	NN	O	O
member	NN	O	O
had	NN	O	O
the	NN	O	O
HLA	NN	O	O
-	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
,	NN	O	O
DR2	NN	O	O
(	NN	O	O
w15	NN	O	O
)	NN	O	O
haplotype	NN	O	O
.	NN	O	O

The	NN	O	O
mother	NN	O	O
had	NN	O	O
seropositive	NN	O	B-Disease
rheumatoid	NN	O	I-Disease
arthritis	NN	O	I-Disease
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
showed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cryoglobulins	NN	O	O
,	NN	O	O
antibodies	NN	O	O
against	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
anticardiolipin	NN	O	O
antibodies	NN	O	O
.	NN	O	O
.	NN	O	O

New	NN	O	O
variant	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
greater	NN	O	O
than	NN	O	O
G	NN	O	O
transversion	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
of	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
their	NN	O	O
mother	NN	O	O
in	NN	O	O
a	NN	O	O
single	NN	O	O
sibship	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
PMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
transversion	NN	O	O
should	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
an	NN	O	O
amino	NN	O	O
acid	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
protein	NN	O	O
but	NN	O	O
it	NN	O	O
does	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
a	NN	O	O
HaeIII	NN	O	O
restriction	NN	O	O
endonuclease	NN	O	O
cleavage	NN	O	O
site	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
concordant	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

One	NN	O	O
-	NN	O	O
hundred	NN	O	O
-	NN	O	O
ten	NN	O	O
unrelated	NN	O	O
X	NN	O	O
chromosomes	NN	O	O
are	NN	O	O
negative	NN	O	O
for	NN	O	O
this	NN	O	O
mutation	NN	O	O
.	NN	O	O

No	NN	O	O
other	NN	O	O
sequence	NN	O	O
defect	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
PLP	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
males	NN	O	O
.	NN	O	O

The	NN	O	O
cause	NN	O	O
of	NN	O	O
disease	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
remains	NN	O	O
unknown	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
association	NN	O	O
between	NN	O	O
this	NN	O	O
rare	NN	O	O
mutation	NN	O	O
and	NN	O	O
PMD	NN	O	B-Disease
is	NN	O	O
intriguing	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
following	NN	O	O
segregation	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
.	NN	O	O
.	NN	O	O

Uncoupling	NN	O	O
of	NN	O	O
hypomyelination	NN	O	B-Disease
and	NN	O	O
glial	NN	O	B-Disease
cell	NN	O	I-Disease
death	NN	O	I-Disease
by	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
gene	NN	O	O
.	NN	O	O

Proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
;	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
30	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
major	NN	O	O
polytopic	NN	O	O
membrane	NN	O	O
protein	NN	O	O
in	NN	O	O
myelin	NN	O	O
but	NN	O	O
its	NN	O	O
cellular	NN	O	O
function	NN	O	O
remains	NN	O	O
obscure	NN	O	O
.	NN	O	O

Neurological	NN	O	O
mutant	NN	O	O
mice	NN	O	O
can	NN	O	O
often	NN	O	O
provide	NN	O	O
model	NN	O	O
systems	NN	O	O
for	NN	O	O
human	NN	O	O
genetic	NN	O	B-Disease
disorders	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
-	NN	O	O
linked	NN	O	O
PLP	NN	O	O
gene	NN	O	O
are	NN	O	O
lethal	NN	O	O
,	NN	O	O
identified	NN	O	O
first	NN	O	O
in	NN	O	O
the	NN	O	O
jimpy	NN	O	O
mouse	NN	O	O
and	NN	O	O
subsequently	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
unexplained	NN	O	O
phenotype	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
includes	NN	O	O
degeneration	NN	O	B-Disease
and	NN	O	I-Disease
premature	NN	O	I-Disease
cell	NN	O	I-Disease
death	NN	O	I-Disease
of	NN	O	I-Disease
oligodendrocytes	NN	O	I-Disease
with	NN	O	O
associated	NN	O	O
hypomyelination	NN	O	B-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
new	NN	O	O
mouse	NN	O	O
mutant	NN	O	O
rumpshaker	NN	O	O
is	NN	O	O
defined	NN	O	O
by	NN	O	O
the	NN	O	O
amino	NN	O	O
-	NN	O	O
acid	NN	O	O
substitution	NN	O	O
Ile	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
Thr	NN	O	O
at	NN	O	O
residue	NN	O	O
186	NN	O	O
in	NN	O	O
a	NN	O	O
membrane	NN	O	O
-	NN	O	O
embedded	NN	O	O
domain	NN	O	O
of	NN	O	O
PLP	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
rumpshaker	NN	O	O
mice	NN	O	O
,	NN	O	O
although	NN	O	O
myelin	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
,	NN	O	O
have	NN	O	O
normal	NN	O	O
longevity	NN	O	O
and	NN	O	O
a	NN	O	O
full	NN	O	O
complement	NN	O	O
of	NN	O	O
morphologically	NN	O	O
normal	NN	O	O
oligodendrocytes	NN	O	O
.	NN	O	O

Hypomyelination	NN	O	B-Disease
can	NN	O	O
thus	NN	O	O
be	NN	O	O
genetically	NN	O	O
separated	NN	O	O
from	NN	O	O
the	NN	O	O
PLP	NN	O	O
-	NN	O	O
dependent	NN	O	O
oligodendrocyte	NN	O	B-Disease
degeneration	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
PLP	NN	O	O
has	NN	O	O
a	NN	O	O
vital	NN	O	O
function	NN	O	O
in	NN	O	O
glial	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
distinct	NN	O	O
from	NN	O	O
its	NN	O	O
later	NN	O	O
role	NN	O	O
in	NN	O	O
myelin	NN	O	O
assembly	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
dichotomy	NN	O	O
of	NN	O	O
action	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
clinical	NN	O	O
spectrum	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
pseudodeficiency	NN	O	O
allele	NN	O	O
common	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
carriers	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
carrier	NN	O	O
screening	NN	O	O
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
beta	NN	O	I-Disease
-	NN	O	I-Disease
hexosaminidase	NN	O	I-Disease
A	NN	O	I-Disease
(	NN	O	O
Hex	NN	O	O
A	NN	O	O
)	NN	O	O
activity	NN	O	O
typically	NN	O	O
results	NN	O	O
in	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
deficient	NN	O	B-Disease
in	NN	O	I-Disease
Hex	NN	O	I-Disease
A	NN	O	I-Disease
activity	NN	O	O
(	NN	O	O
i	NN	O	O
.	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
pseudodeficient	NN	O	O
)	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
biochemical	NN	O	O
tests	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
of	NN	O	O
one	NN	O	O
pseudodeficient	NN	O	O
subject	NN	O	O
and	NN	O	O
identified	NN	O	O
both	NN	O	O
a	NN	O	O
C739	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
substitution	NN	O	O
that	NN	O	O
changes	NN	O	O
Arg247	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Trp	NN	O	O
on	NN	O	O
one	NN	O	O
allele	NN	O	O
and	NN	O	O
a	NN	O	O
previously	NN	O	O
identified	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
mutation	NN	O	O
on	NN	O	O
the	NN	O	O
second	NN	O	O
allele	NN	O	O
.	NN	O	O

Six	NN	O	O
additional	NN	O	O
pseudodeficient	NN	O	O
subjects	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
C739	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
mutation	NN	O	O
.	NN	O	O

This	NN	O	O
allele	NN	O	O
accounted	NN	O	O
for	NN	O	O
32	NN	O	O
%	NN	O	O
(	NN	O	O
20	NN	O	O
/	NN	O	O
62	NN	O	O
)	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
enzyme	NN	O	O
-	NN	O	O
defined	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
carriers	NN	O	O
but	NN	O	O
for	NN	O	O
none	NN	O	O
of	NN	O	O
36	NN	O	O
Jewish	NN	O	O
enzyme	NN	O	O
-	NN	O	O
defined	NN	O	O
carriers	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
one	NN	O	O
of	NN	O	O
three	NN	O	O
known	NN	O	O
mutations	NN	O	O
common	NN	O	O
to	NN	O	O
this	NN	O	O
group	NN	O	O
.	NN	O	O

The	NN	O	O
C739	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
allele	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
"	NN	O	O
true	NN	O	O
"	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
allele	NN	O	O
,	NN	O	O
causes	NN	O	O
Hex	NN	O	O
A	NN	O	O
pseudodeficiency	NN	O	O
.	NN	O	O

Given	NN	O	O
both	NN	O	O
the	NN	O	O
large	NN	O	O
proportion	NN	O	O
of	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
carriers	NN	O	O
with	NN	O	O
this	NN	O	O
allele	NN	O	O
and	NN	O	O
that	NN	O	O
standard	NN	O	O
biochemical	NN	O	O
screening	NN	O	O
cannot	NN	O	O
differentiate	NN	O	O
between	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
the	NN	O	O
C739	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
mutations	NN	O	O
and	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
carriers	NN	O	O
,	NN	O	O
DNA	NN	O	O
testing	NN	O	O
for	NN	O	O
this	NN	O	O
mutation	NN	O	O
in	NN	O	O
at	NN	O	O
-	NN	O	O
risk	NN	O	O
couples	NN	O	O
is	NN	O	O
essential	NN	O	O
.	NN	O	O

Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
:	NN	O	O
detection	NN	O	O
of	NN	O	O
mutations	NN	O	O
Thr181	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Pro	NN	O	O
and	NN	O	O
Leu223	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Pro	NN	O	O
in	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
history	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
presented	NN	O	O
for	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
,	NN	O	O
requesting	NN	O	O
carrier	NN	O	O
detection	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

RFLP	NN	O	O
analysis	NN	O	O
using	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
probe	NN	O	O
was	NN	O	O
uninformative	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

A	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
on	NN	O	O
a	NN	O	O
chorionic	NN	O	O
villus	NN	O	O
sample	NN	O	O
(	NN	O	O
CVS	NN	O	O
)	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
using	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
variant	NN	O	O
in	NN	O	O
exon	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
fetus	NN	O	O
was	NN	O	O
predicted	NN	O	O
to	NN	O	O
be	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
exon	NN	O	O
from	NN	O	O
the	NN	O	O
CVS	NN	O	O
,	NN	O	O
the	NN	O	O
predicted	NN	O	O
-	NN	O	O
carrier	NN	O	O
mother	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
obligate	NN	O	O
-	NN	O	O
carrier	NN	O	O
grandmother	NN	O	O
revealed	NN	O	O
an	NN	O	O
A	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
C	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
541	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
women	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
fetus	NN	O	O
.	NN	O	O

As	NN	O	O
this	NN	O	O
change	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
Thr	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
Pro	NN	O	O
change	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
181	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
predicted	NN	O	O
to	NN	O	O
be	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
transmembrane	NN	O	O
segment	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
this	NN	O	O
was	NN	O	O
the	NN	O	O
mutation	NN	O	O
causing	NN	O	O
the	NN	O	O
disease	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
second	NN	O	O
family	NN	O	O
,	NN	O	O
an	NN	O	O
exon	NN	O	O
5	NN	O	O
variant	NN	O	O
band	NN	O	O
pattern	NN	O	O
on	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
sequencing	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
T	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
C	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
668	NN	O	O
.	NN	O	O

This	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
Leu	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
Pro	NN	O	O
change	NN	O	O
in	NN	O	O
a	NN	O	O
carrier	NN	O	O
mother	NN	O	O
and	NN	O	O
in	NN	O	O
her	NN	O	O
two	NN	O	O
affected	NN	O	O
sons	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
further	NN	O	O
examples	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
PLP	NN	O	O
that	NN	O	O
cause	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
and	NN	O	O
illustrate	NN	O	O
the	NN	O	O
value	NN	O	O
of	NN	O	O
SSCP	NN	O	O
in	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
.	NN	O	O
.	NN	O	O

von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
type	NN	O	I-Disease
B	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
selectively	NN	O	O
abolishes	NN	O	O
ristocetin	NN	O	O
-	NN	O	O
induced	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
binding	NN	O	O
to	NN	O	O
platelet	NN	O	O
glycoprotein	NN	O	O
Ib	NN	O	O
.	NN	O	O

von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
(	NN	O	O
vWF	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
multimeric	NN	O	O
glycoprotein	NN	O	O
that	NN	O	O
mediates	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
platelets	NN	O	O
to	NN	O	O
the	NN	O	O
subendothelium	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
platelet	NN	O	O
glycoprotein	NN	O	O
Ib	NN	O	O
.	NN	O	O

For	NN	O	O
human	NN	O	O
vWF	NN	O	O
,	NN	O	O
this	NN	O	O
interaction	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
the	NN	O	O
antibiotic	NN	O	O
ristocetin	NN	O	O
or	NN	O	O
the	NN	O	O
snake	NN	O	O
venom	NN	O	O
protein	NN	O	O
botrocetin	NN	O	O
.	NN	O	O

A	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
Gly	NN	O	O
-	NN	O	O
561	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Ser	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
within	NN	O	O
the	NN	O	O
proposed	NN	O	O
glycoprotein	NN	O	O
Ib	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
vWF	NN	O	O
in	NN	O	O
the	NN	O	O
proband	NN	O	O
with	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
type	NN	O	I-Disease
B	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
unique	NN	O	O
variant	NN	O	O
characterized	NN	O	O
by	NN	O	O
no	NN	O	O
ristocetin	NN	O	O
-	NN	O	O
induced	NN	O	O
,	NN	O	O
but	NN	O	O
normal	NN	O	O
botrocetin	NN	O	O
-	NN	O	O
induced	NN	O	O
,	NN	O	O
binding	NN	O	O
to	NN	O	O
glycoprotein	NN	O	O
Ib	NN	O	O
.	NN	O	O

The	NN	O	O
corresponding	NN	O	O
mutant	NN	O	O
recombinant	NN	O	O
protein	NN	O	O
,	NN	O	O
rvWF	NN	O	O
(	NN	O	O
G561S	NN	O	O
)	NN	O	O
,	NN	O	O
formed	NN	O	O
normal	NN	O	O
multimers	NN	O	O
and	NN	O	O
exhibited	NN	O	O
the	NN	O	O
same	NN	O	O
functional	NN	O	O
defect	NN	O	O
as	NN	O	O
the	NN	O	O
patients	NN	O	O
plasma	NN	O	O
vWF	NN	O	O
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
causes	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
type	NN	O	I-Disease
B	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
botrocetin	NN	O	O
and	NN	O	O
ristocetin	NN	O	O
cofactor	NN	O	O
activities	NN	O	O
of	NN	O	O
vWF	NN	O	O
can	NN	O	O
be	NN	O	O
dissociated	NN	O	O
by	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
and	NN	O	O
confirm	NN	O	O
that	NN	O	O
these	NN	O	O
mediators	NN	O	O
promote	NN	O	O
vWF	NN	O	O
binding	NN	O	O
to	NN	O	O
platelets	NN	O	O
by	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

The	NN	O	O
normal	NN	O	O
botrocetin	NN	O	O
-	NN	O	O
induced	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
defective	NN	O	O
ristocetin	NN	O	O
-	NN	O	O
induced	NN	O	O
binding	NN	O	O
of	NN	O	O
rvWF	NN	O	O
(	NN	O	O
G561S	NN	O	O
)	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
primary	NN	O	O
defect	NN	O	O
in	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
type	NN	O	I-Disease
B	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
a	NN	O	O
failure	NN	O	O
of	NN	O	O
normal	NN	O	O
allosteric	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glycoprotein	NN	O	O
Ib	NN	O	O
binding	NN	O	O
function	NN	O	O
of	NN	O	O
vWF	NN	O	O
.	NN	O	O
.	NN	O	O

Typical	NN	O	O
and	NN	O	O
partial	NN	O	O
cat	NN	O	B-Disease
eye	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
marker	NN	O	O
chromosome	NN	O	O
by	NN	O	O
FISH	NN	O	O
.	NN	O	O

Three	NN	O	O
children	NN	O	O
are	NN	O	O
reported	NN	O	O
with	NN	O	O
typical	NN	O	O
cat	NN	O	B-Disease
eye	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
CES	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
three	NN	O	O
more	NN	O	O
children	NN	O	O
with	NN	O	O
partial	NN	O	O
CES	NN	O	B-Disease
because	NN	O	O
of	NN	O	O
absence	NN	O	O
of	NN	O	O
coloboma	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
supernumerary	NN	O	O
marker	NN	O	O
chromosome	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
FISH	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
genomic	NN	O	O
library	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
a	NN	O	O
centromeric	NN	O	O
and	NN	O	O
particularly	NN	O	O
a	NN	O	O
cosmid	NN	O	O
probe	NN	O	O
of	NN	O	O
22q11	NN	O	O
,	NN	O	O
partial	NN	O	O
tetrasomy	NN	O	B-Disease
was	NN	O	O
shown	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
.	NN	O	O
.	NN	O	O

[	NN	O	O
Genetic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
of	NN	O	O
G6PD	NN	O	O
in	NN	O	O
the	NN	O	O
Shekii	NN	O	O
district	NN	O	O
of	NN	O	O
Azerbaijan	NN	O	O
]	NN	O	O

Examination	NN	O	O
on	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
349	NN	O	O
patients	NN	O	O
of	NN	O	O
Shekii	NN	O	O
district	NN	O	O
hospital	NN	O	O
(	NN	O	O
Azerbaijan	NN	O	O
)	NN	O	O
revealed	NN	O	O
16	NN	O	O
hemi	NN	O	O
-	NN	O	O
,	NN	O	O
4	NN	O	O
homo	NN	O	O
-	NN	O	O
and	NN	O	O
9	NN	O	O
heterozygotic	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
defect	NN	O	O
.	NN	O	O

Gd	NN	O	O
-	NN	O	O
frequency	NN	O	O
,	NN	O	O
calculated	NN	O	O
from	NN	O	O
the	NN	O	O
data	NN	O	O
obtained	NN	O	O
(	NN	O	O
7	NN	O	O
.	NN	O	O
7	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
compared	NN	O	O
to	NN	O	O
neighbouring	NN	O	O
regions	NN	O	O
frequencies	NN	O	O
(	NN	O	O
6	NN	O	O
-	NN	O	O
30	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Carriers	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
are	NN	O	O
residents	NN	O	O
of	NN	O	O
11	NN	O	O
villages	NN	O	O
located	NN	O	O
in	NN	O	O
Alasani	NN	O	O
-	NN	O	O
Aphtalan	NN	O	O
valley	NN	O	O
,	NN	O	O
highly	NN	O	O
endemic	NN	O	O
with	NN	O	O
malaria	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
past	NN	O	O
;	NN	O	O
nearly	NN	O	O
all	NN	O	O
marriages	NN	O	O
are	NN	O	O
endogamic	NN	O	O
.	NN	O	O

Physico	NN	O	O
-	NN	O	O
chemical	NN	O	O
and	NN	O	O
kinetic	NN	O	O
study	NN	O	O
of	NN	O	O
10	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
G6PD	NN	O	O
,	NN	O	O
according	NN	O	O
to	NN	O	O
WHO	NN	O	O
program	NN	O	O
,	NN	O	O
led	NN	O	O
to	NN	O	O
identification	NN	O	O
of	NN	O	O
5	NN	O	O
variants	NN	O	O
of	NN	O	O
the	NN	O	O
II	NN	O	O
class	NN	O	O
(	NN	O	O
Shekii	NN	O	O
,	NN	O	O
Bideiz	NN	O	O
,	NN	O	O
Shirin	NN	O	O
-	NN	O	O
Bulakh	NN	O	O
,	NN	O	O
Okhut	NN	O	O
I	NN	O	O
and	NN	O	O
Zakataly	NN	O	O
)	NN	O	O
and	NN	O	O
2	NN	O	O
variants	NN	O	O
of	NN	O	O
the	NN	O	O
III	NN	O	O
class	NN	O	O
(	NN	O	O
Okhut	NN	O	O
II	NN	O	O
and	NN	O	O
Martinique	NN	O	O
-	NN	O	O
like	NN	O	O
)	NN	O	O
.	NN	O	O

Resemblance	NN	O	O
of	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
variants	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
and	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
erythrocyte	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
permit	NN	O	O
to	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
parental	NN	O	O
mutant	NN	O	O
G6PD	NN	O	O
allele	NN	O	O
distributed	NN	O	O
in	NN	O	O
this	NN	O	O
area	NN	O	O
.	NN	O	O

Craniofrontonasal	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
nine	NN	O	O
patients	NN	O	O
with	NN	O	O
craniofrontonasal	NN	O	B-Disease
dysplasia	NN	O	I-Disease
(	NN	O	O
CFND	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Seven	NN	O	O
classical	NN	O	O
cases	NN	O	O
had	NN	O	O
facial	NN	O	O
features	NN	O	O
suggestive	NN	O	O
of	NN	O	O
frontonasal	NN	O	B-Disease
dysplasia	NN	O	I-Disease
and	NN	O	O
coronal	NN	O	B-Disease
craniosynostosis	NN	O	I-Disease
.	NN	O	O

Extracranial	NN	O	B-Disease
abnormalities	NN	O	I-Disease
such	NN	O	O
as	NN	O	O
brittle	NN	O	B-Disease
nails	NN	O	I-Disease
with	NN	O	I-Disease
prominent	NN	O	I-Disease
longitudinal	NN	O	I-Disease
grooves	NN	O	I-Disease
or	NN	O	O
syndactyly	NN	O	B-Disease
of	NN	O	I-Disease
fingers	NN	O	I-Disease
and	NN	O	I-Disease
toes	NN	O	I-Disease
were	NN	O	O
observed	NN	O	O
in	NN	O	O
individual	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
families	NN	O	O
the	NN	O	O
father	NN	O	O
of	NN	O	O
classical	NN	O	O
cases	NN	O	O
showed	NN	O	O
a	NN	O	O
milder	NN	O	O
pattern	NN	O	O
of	NN	O	O
abnormalities	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
2	NN	O	O
-	NN	O	O
to	NN	O	O
13	NN	O	O
-	NN	O	O
year	NN	O	O
follow	NN	O	O
-	NN	O	O
up	NN	O	O
on	NN	O	O
our	NN	O	O
patients	NN	O	O
.	NN	O	O

Hypotonia	NN	O	B-Disease
and	NN	O	O
laxity	NN	O	B-Disease
of	NN	O	I-Disease
joints	NN	O	I-Disease
are	NN	O	O
common	NN	O	O
and	NN	O	O
may	NN	O	O
necessitate	NN	O	O
supportive	NN	O	O
measures	NN	O	O
.	NN	O	O

Mild	NN	O	O
developmental	NN	O	B-Disease
delay	NN	O	I-Disease
was	NN	O	O
noted	NN	O	O
in	NN	O	O
three	NN	O	O
out	NN	O	O
of	NN	O	O
six	NN	O	O
classical	NN	O	O
cases	NN	O	O
studied	NN	O	O
in	NN	O	O
detail	NN	O	O
.	NN	O	O

Unlike	NN	O	O
almost	NN	O	O
all	NN	O	O
other	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disorders	NN	O	I-Disease
,	NN	O	O
clinical	NN	O	O
expression	NN	O	O
in	NN	O	O
CFND	NN	O	B-Disease
is	NN	O	O
generally	NN	O	O
much	NN	O	O
more	NN	O	O
severe	NN	O	O
in	NN	O	O
females	NN	O	O
than	NN	O	O
in	NN	O	O
males	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
previous	NN	O	O
reports	NN	O	O
of	NN	O	O
this	NN	O	O
condition	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
our	NN	O	O
severely	NN	O	O
affected	NN	O	O
cases	NN	O	O
is	NN	O	O
a	NN	O	O
male	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
intron	NN	O	O
7	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
transition	NN	O	O
:	NN	O	O
a	NN	O	O
second	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
mutation	NN	O	O
in	NN	O	O
French	NN	O	O
Canada	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
hexosaminidase	NN	O	O
A	NN	O	O
(	NN	O	O
HEXA	NN	O	O
)	NN	O	O
gene	NN	O	O
which	NN	O	O
cause	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
elevated	NN	O	O
frequency	NN	O	O
in	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
and	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
populations	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
novel	NN	O	O
TSD	NN	O	B-Disease
allele	NN	O	O
in	NN	O	O
the	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
population	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
infantile	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
+	NN	O	O
1	NN	O	O
position	NN	O	O
of	NN	O	O
intron	NN	O	O
7	NN	O	O
,	NN	O	O
abolishes	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
.	NN	O	O

Cultured	NN	O	O
human	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
a	NN	O	O
compound	NN	O	O
heterozygote	NN	O	O
for	NN	O	O
this	NN	O	O
transition	NN	O	O
(	NN	O	O
and	NN	O	O
for	NN	O	O
a	NN	O	O
deletion	NN	O	O
mutation	NN	O	O
)	NN	O	O
produce	NN	O	O
no	NN	O	O
detectable	NN	O	O
HEXA	NN	O	O
mRNA	NN	O	O
.	NN	O	O

The	NN	O	O
intron	NN	O	O
7	NN	O	O
+	NN	O	O
1	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
base	NN	O	O
adjacent	NN	O	O
to	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
TSD	NN	O	B-Disease
mutation	NN	O	O
(	NN	O	O
G805A	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
mutations	NN	O	O
a	NN	O	O
restriction	NN	O	O
site	NN	O	O
for	NN	O	O
the	NN	O	O
endonuclease	NN	O	O
EcoRII	NN	O	O
is	NN	O	O
abolished	NN	O	O
.	NN	O	O

Unambiguous	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
requires	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
hybridization	NN	O	O
to	NN	O	O
distinguish	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
.	NN	O	O

The	NN	O	O
intron	NN	O	O
7	NN	O	O
+	NN	O	O
1	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
three	NN	O	O
unrelated	NN	O	O
families	NN	O	O
.	NN	O	O

Obligate	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
the	NN	O	O
intron	NN	O	O
7	NN	O	O
+	NN	O	O
1	NN	O	O
mutation	NN	O	O
were	NN	O	O
born	NN	O	O
in	NN	O	O
the	NN	O	O
Saguenay	NN	O	O
-	NN	O	O
Lac	NN	O	O
-	NN	O	O
St	NN	O	O
-	NN	O	O
Jean	NN	O	O
region	NN	O	O
of	NN	O	O
Quebec	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
recent	NN	O	O
ancestors	NN	O	O
common	NN	O	O
to	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
were	NN	O	O
from	NN	O	O
the	NN	O	O
Charlevoix	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
province	NN	O	O
of	NN	O	O
Quebec	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
thus	NN	O	O
has	NN	O	O
a	NN	O	O
different	NN	O	O
geographic	NN	O	O
centre	NN	O	O
of	NN	O	O
diffusion	NN	O	O
and	NN	O	O
is	NN	O	O
probably	NN	O	O
less	NN	O	O
common	NN	O	O
than	NN	O	O
the	NN	O	O
exon	NN	O	O
1	NN	O	O
deletion	NN	O	O
TSD	NN	O	B-Disease
mutation	NN	O	O
in	NN	O	O
French	NN	O	O
Canadians	NN	O	O
.	NN	O	O

Neither	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
France	NN	O	O
,	NN	O	O
the	NN	O	O
ancestral	NN	O	O
homeland	NN	O	O
of	NN	O	O
French	NN	O	O
Canada	NN	O	O
.	NN	O	O
.	NN	O	O

Assignment	NN	O	O
of	NN	O	O
the	NN	O	O
aspartylglucosaminidase	NN	O	O
gene	NN	O	O
(	NN	O	O
AGA	NN	O	O
)	NN	O	O
to	NN	O	O
4q33	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q35	NN	O	O
based	NN	O	O
on	NN	O	O
decreased	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
girl	NN	O	O
with	NN	O	O
a	NN	O	O
46	NN	O	O
,	NN	O	O
XX	NN	O	O
,	NN	O	O
del	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
(	NN	O	O
q33	NN	O	O
)	NN	O	O
karyotype	NN	O	O
.	NN	O	O

Aspartylglucosaminuria	NN	O	B-Disease
(	NN	O	O
AGU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
recessive	NN	O	O
autosomally	NN	O	O
inherited	NN	O	O
lysosomal	NN	O	B-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
enzyme	NN	O	I-Disease
aspartylglucosaminidase	NN	O	I-Disease
(	NN	O	O
AGA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
structural	NN	O	O
gene	NN	O	O
for	NN	O	O
this	NN	O	O
human	NN	O	O
enzyme	NN	O	O
(	NN	O	O
AGA	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
assigned	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
4q21	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
the	NN	O	O
AGA	NN	O	O
activity	NN	O	O
in	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
of	NN	O	O
a	NN	O	O
girl	NN	O	O
with	NN	O	O
a	NN	O	O
46	NN	O	O
,	NN	O	O
XX	NN	O	O
,	NN	O	O
del	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
(	NN	O	O
q33	NN	O	O
)	NN	O	O
karyotype	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
girl	NN	O	O
is	NN	O	O
a	NN	O	O
hemizygote	NN	O	O
for	NN	O	O
AGA	NN	O	O
,	NN	O	O
permitting	NN	O	O
the	NN	O	O
assignment	NN	O	O
of	NN	O	O
human	NN	O	O
AGA	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
4q33	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O
.	NN	O	O

Resolution	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
loci	NN	O	O
for	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
AN1	NN	O	O
and	NN	O	O
AN2	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
single	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
11p13	NN	O	O
.	NN	O	O

Two	NN	O	O
distinct	NN	O	O
loci	NN	O	O
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
for	NN	O	O
aniridia	NN	O	B-Disease
;	NN	O	O
AN1	NN	O	O
for	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
on	NN	O	O
chromosome	NN	O	O
2p	NN	O	O
and	NN	O	O
AN2	NN	O	O
for	NN	O	O
the	NN	O	O
aniridia	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
WAGR	NN	O	B-Disease
contiguous	NN	O	I-Disease
gene	NN	O	I-Disease
syndrome	NN	O	I-Disease
on	NN	O	O
chromosome	NN	O	O
11p13	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
the	NN	O	O
kindred	NN	O	O
segregating	NN	O	O
for	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
which	NN	O	O
suggested	NN	O	O
linkage	NN	O	O
to	NN	O	O
acid	NN	O	O
phosphatase	NN	O	O
-	NN	O	O
1	NN	O	O
(	NN	O	O
ACP1	NN	O	O
)	NN	O	O
and	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
assignment	NN	O	O
of	NN	O	O
the	NN	O	O
AN1	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
2p	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
updated	NN	O	O
and	NN	O	O
expanded	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
between	NN	O	O
the	NN	O	O
aniridia	NN	O	B-Disease
phenotype	NN	O	O
and	NN	O	O
ACP1	NN	O	O
does	NN	O	O
not	NN	O	O
support	NN	O	O
the	NN	O	O
original	NN	O	O
linkage	NN	O	O
results	NN	O	O
,	NN	O	O
excluding	NN	O	O
linkage	NN	O	O
up	NN	O	O
to	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

17	NN	O	O
with	NN	O	O
Z	NN	O	O
=	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O

Tests	NN	O	O
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
other	NN	O	O
chromosome	NN	O	O
2p	NN	O	O
markers	NN	O	O
.	NN	O	O

APOB	NN	O	O
,	NN	O	O
D2S71	NN	O	O
,	NN	O	O
D2S5	NN	O	O
,	NN	O	O
and	NN	O	O
D2S1	NN	O	O
,	NN	O	O
also	NN	O	O
excluded	NN	O	O
linkage	NN	O	O
to	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O

Markers	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
chromosome	NN	O	O
11p13	NN	O	O
region	NN	O	O
were	NN	O	O
then	NN	O	O
analyzed	NN	O	O
in	NN	O	O
this	NN	O	O
aniridia	NN	O	B-Disease
family	NN	O	O
.	NN	O	O

Two	NN	O	O
RFLPs	NN	O	O
at	NN	O	O
the	NN	O	O
D11S323	NN	O	O
locus	NN	O	O
give	NN	O	O
significant	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
.	NN	O	O

The	NN	O	O
PvuII	NN	O	O
polymorphism	NN	O	O
detected	NN	O	O
by	NN	O	O
probe	NN	O	O
p5S1	NN	O	O
.	NN	O	O

6	NN	O	O
detects	NN	O	O
no	NN	O	O
recombinants	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
Z	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O

97	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

00	NN	O	O
00	NN	O	O
.	NN	O	O

The	NN	O	O
HaeIII	NN	O	O
polymorphism	NN	O	O
detected	NN	O	O
by	NN	O	O
the	NN	O	O
probe	NN	O	O
p5BE1	NN	O	O
.	NN	O	O

2	NN	O	O
gives	NN	O	O
a	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
Z	NN	O	O
=	NN	O	O
2	NN	O	O
.	NN	O	O

57	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

00	NN	O	O
00	NN	O	O
.	NN	O	O

Locus	NN	O	O
D11S325	NN	O	O
gives	NN	O	O
a	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
Z	NN	O	O
=	NN	O	O
1	NN	O	O
.	NN	O	O

53	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

00	NN	O	O
00	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
locus	NN	O	O
for	NN	O	O
aniridia	NN	O	B-Disease
(	NN	O	O
AN1	NN	O	O
)	NN	O	O
on	NN	O	O
chromosome	NN	O	O
2p	NN	O	O
has	NN	O	O
been	NN	O	O
misassigned	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
family	NN	O	O
is	NN	O	O
segregating	NN	O	O
for	NN	O	O
an	NN	O	O
aniridia	NN	O	B-Disease
mutation	NN	O	O
linked	NN	O	O
to	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
11p13	NN	O	O
region	NN	O	O
.	NN	O	O

Fatal	NN	O	O
pyoderma	NN	O	B-Disease
gangrenosum	NN	O	I-Disease
in	NN	O	O
association	NN	O	O
with	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
pyoderma	NN	O	B-Disease
gangrenosum	NN	O	I-Disease
(	NN	O	O
PG	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
often	NN	O	O
associated	NN	O	O
with	NN	O	O
systemic	NN	O	B-Disease
diseases	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
complement	NN	O	I-Disease
deficiencies	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
describe	NN	O	O
an	NN	O	O
aggressive	NN	O	O
and	NN	O	O
ultimately	NN	O	O
fatal	NN	O	O
case	NN	O	O
of	NN	O	O
PG	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
congenital	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Deficiencies	NN	O	B-Disease
of	NN	O	I-Disease
C7	NN	O	I-Disease
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
neutrophil	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
phagocytosis	NN	O	O
,	NN	O	O
and	NN	O	O
opsonization	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
immunologic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
described	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
PG	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
patients	NN	O	O
decreased	NN	O	O
complement	NN	O	O
level	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
directly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
PG	NN	O	B-Disease
,	NN	O	O
may	NN	O	O
have	NN	O	O
contributed	NN	O	O
to	NN	O	O
the	NN	O	O
aggressive	NN	O	O
nature	NN	O	O
of	NN	O	O
her	NN	O	O
disease	NN	O	O
.	NN	O	O
.	NN	O	O

Diverse	NN	O	O
point	NN	O	O
mutations	NN	O	O
result	NN	O	O
in	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
PHOSPHATE	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
;	NN	O	I-Disease
EC	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
49	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
human	NN	O	O
enzymopathy	NN	O	B-Disease
,	NN	O	O
affecting	NN	O	O
more	NN	O	O
than	NN	O	O
200	NN	O	O
million	NN	O	O
people	NN	O	O
worldwide	NN	O	O
.	NN	O	O

Although	NN	O	O
greater	NN	O	O
than	NN	O	O
400	NN	O	O
variants	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
based	NN	O	O
on	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
criteria	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
these	NN	O	O
G6PD	NN	O	B-Disease
deficiencies	NN	O	I-Disease
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
human	NN	O	O
G6PD	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
,	NN	O	O
which	NN	O	O
enables	NN	O	O
us	NN	O	O
to	NN	O	O
examine	NN	O	O
directly	NN	O	O
the	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	O
at	NN	O	O
the	NN	O	O
DNA	NN	O	O
level	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
past	NN	O	O
10	NN	O	O
years	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
G6PD	NN	O	O
activity	NN	O	O
in	NN	O	O
21	NN	O	O
,	NN	O	O
271	NN	O	O
newborn	NN	O	O
Chinese	NN	O	O
infants	NN	O	O
(	NN	O	O
11	NN	O	O
,	NN	O	O
400	NN	O	O
males	NN	O	O
and	NN	O	O
9	NN	O	O
,	NN	O	O
871	NN	O	O
females	NN	O	O
)	NN	O	O
and	NN	O	O
identified	NN	O	O
314	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
males	NN	O	O
and	NN	O	O
246	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
females	NN	O	O
having	NN	O	O
low	NN	O	O
G6PD	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
G6PD	NN	O	O
gene	NN	O	O
from	NN	O	O
10	NN	O	O
randomly	NN	O	O
selected	NN	O	O
affected	NN	O	O
individuals	NN	O	O
and	NN	O	O
their	NN	O	O
relatives	NN	O	O
was	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
amplified	NN	O	O
,	NN	O	O
subcloned	NN	O	O
,	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
four	NN	O	O
types	NN	O	O
of	NN	O	O
mutation	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
G6PD	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
(	NN	O	O
487	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
an	NN	O	O
affected	NN	O	O
Chinese	NN	O	O
with	NN	O	O
a	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
487	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
(	NN	O	O
163	NN	O	O
)	NN	O	O
Gly	NN	O	O
to	NN	O	O
Ser	NN	O	O
substitution	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
(	NN	O	O
493	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
G	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
that	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
any	NN	O	O
other	NN	O	O
ethnic	NN	O	O
group	NN	O	O
and	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
two	NN	O	O
affected	NN	O	O
Chinese	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
causes	NN	O	O
an	NN	O	O
A	NN	O	O
to	NN	O	O
G	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
493	NN	O	O
,	NN	O	O
producing	NN	O	O
an	NN	O	O
(	NN	O	O
165	NN	O	O
)	NN	O	O
Asn	NN	O	O
to	NN	O	O
Asp	NN	O	O
substitution	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
487	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
and	NN	O	O
493	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
G	NN	O	O
mutations	NN	O	O
create	NN	O	O
Alu	NN	O	O
I	NN	O	O
and	NN	O	O
Ava	NN	O	O
II	NN	O	O
recognition	NN	O	O
sites	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
which	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
rapidly	NN	O	O
detect	NN	O	O
these	NN	O	O
two	NN	O	O
mutations	NN	O	O
by	NN	O	O
PCR	NN	O	O
/	NN	O	O
restriction	NN	O	O
enzyme	NN	O	O
(	NN	O	O
RE	NN	O	O
)	NN	O	O
digestion	NN	O	O
method	NN	O	O
.	NN	O	O

The	NN	O	O
third	NN	O	O
mutation	NN	O	O
(	NN	O	O
1376	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
four	NN	O	O
affected	NN	O	O
Chinese	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
causes	NN	O	O
a	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
1376	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
(	NN	O	O
459	NN	O	O
)	NN	O	O
Arg	NN	O	O
to	NN	O	O
Leu	NN	O	O
substitution	NN	O	O
.	NN	O	O

The	NN	O	O
1376	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
mutation	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
dominant	NN	O	O
allele	NN	O	O
that	NN	O	O
causes	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Taiwan	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
two	NN	O	O
affected	NN	O	O
Chinese	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
having	NN	O	O
the	NN	O	O
fourth	NN	O	O
mutation	NN	O	O
(	NN	O	O
1388	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
causes	NN	O	O
a	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
change	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1388	NN	O	O
that	NN	O	O
produces	NN	O	O
an	NN	O	O
(	NN	O	O
463	NN	O	O
)	NN	O	O
Arg	NN	O	O
to	NN	O	O
His	NN	O	O
substitution	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
provide	NN	O	O
the	NN	O	O
direct	NN	O	O
proof	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
Chinese	NN	O	O
populations	NN	O	O
of	NN	O	O
Taiwan	NN	O	O
and	NN	O	O
the	NN	O	O
PCR	NN	O	O
/	NN	O	O
RE	NN	O	O
digestion	NN	O	O
method	NN	O	O
is	NN	O	O
suitable	NN	O	O
for	NN	O	O
simultaneous	NN	O	O
detection	NN	O	O
of	NN	O	O
the	NN	O	O
487	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
and	NN	O	O
493	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
G	NN	O	O
mutations	NN	O	O
.	NN	O	O

An	NN	O	O
error	NN	O	O
in	NN	O	O
dystrophin	NN	O	O
mRNA	NN	O	O
processing	NN	O	O
in	NN	O	O
golden	NN	O	B-Disease
retriever	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
an	NN	O	O
animal	NN	O	O
homologue	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Golden	NN	O	B-Disease
retriever	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
GRMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
spontaneous	NN	O	O
,	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
,	NN	O	O
progressively	NN	O	O
fatal	NN	O	O
disease	NN	O	O
of	NN	O	O
dogs	NN	O	O
and	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
homologue	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Two	NN	O	O
-	NN	O	O
thirds	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
carry	NN	O	O
detectable	NN	O	O
deletions	NN	O	O
in	NN	O	O
their	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
defect	NN	O	O
underlying	NN	O	O
the	NN	O	O
remaining	NN	O	O
one	NN	O	O
-	NN	O	O
third	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
is	NN	O	O
undetermined	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
canine	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
GRMD	NN	O	B-Disease
dogs	NN	O	O
has	NN	O	O
failed	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
any	NN	O	O
detectable	NN	O	O
loss	NN	O	O
of	NN	O	O
exons	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
RNA	NN	O	O
processing	NN	O	O
error	NN	O	O
in	NN	O	O
GRMD	NN	O	B-Disease
that	NN	O	O
results	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
consensus	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
6	NN	O	O
.	NN	O	O

The	NN	O	O
seventh	NN	O	O
exon	NN	O	O
is	NN	O	O
then	NN	O	O
skipped	NN	O	O
,	NN	O	O
which	NN	O	O
predicts	NN	O	O
a	NN	O	O
termination	NN	O	O
of	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
reading	NN	O	O
frame	NN	O	O
within	NN	O	O
its	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
domain	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
dystrophin	NN	O	B-Disease
deficiency	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
.	NN	O	O
.	NN	O	O

Type	NN	O	B-Disease
I	NN	O	I-Disease
human	NN	O	I-Disease
complement	NN	O	I-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
gene	NN	O	O
deletion	NN	O	O
causes	NN	O	O
skipping	NN	O	O
of	NN	O	O
exon	NN	O	O
6	NN	O	O
during	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
.	NN	O	O

Two	NN	O	O
variants	NN	O	O
of	NN	O	O
a	NN	O	O
genetic	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
protein	NN	O	I-Disease
C2	NN	O	I-Disease
(	NN	O	O
C2D	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
previously	NN	O	O
identified	NN	O	O
.	NN	O	O

No	NN	O	O
C2	NN	O	O
protein	NN	O	O
translation	NN	O	O
is	NN	O	O
detected	NN	O	O
in	NN	O	O
type	NN	O	O
I	NN	O	O
deficiency	NN	O	O
,	NN	O	O
while	NN	O	O
type	NN	O	O
II	NN	O	O
deficiency	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
selective	NN	O	O
block	NN	O	O
in	NN	O	O
C2	NN	O	O
secretion	NN	O	O
.	NN	O	O

Type	NN	O	B-Disease
I	NN	O	I-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
described	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
C2	NN	O	O
null	NN	O	O
allele	NN	O	O
(	NN	O	O
C2Q0	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
haplotype	NN	O	O
/	NN	O	O
complotype	NN	O	O
HLA	NN	O	O
-	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
,	NN	O	O
C2Q0	NN	O	O
,	NN	O	O
BfS	NN	O	O
,	NN	O	O
C4A4	NN	O	O
,	NN	O	O
C4B2	NN	O	O
,	NN	O	O
Drw2	NN	O	O
;	NN	O	O
this	NN	O	O
extended	NN	O	O
haplotype	NN	O	O
occurs	NN	O	O
in	NN	O	O
over	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
individuals	NN	O	O
(	NN	O	O
common	NN	O	O
complotype	NN	O	O
/	NN	O	O
haplotype	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
C2	NN	O	O
gene	NN	O	O
and	NN	O	O
cDNA	NN	O	O
were	NN	O	O
characterized	NN	O	O
from	NN	O	O
a	NN	O	O
homozygous	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
C2	NN	O	I-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
individual	NN	O	O
with	NN	O	O
the	NN	O	O
common	NN	O	O
associated	NN	O	O
haplotype	NN	O	O
/	NN	O	O
complotype	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
type	NN	O	O
I	NN	O	O
C2Q0	NN	O	O
gene	NN	O	O
,	NN	O	O
beginning	NN	O	O
9	NN	O	O
base	NN	O	O
pairs	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
3	NN	O	O
-	NN	O	O
end	NN	O	O
of	NN	O	O
exon	NN	O	O
6	NN	O	O
,	NN	O	O
that	NN	O	O
generates	NN	O	O
a	NN	O	O
C2	NN	O	O
transcript	NN	O	O
with	NN	O	O
a	NN	O	O
complete	NN	O	O
deletion	NN	O	O
of	NN	O	O
exon	NN	O	O
6	NN	O	O
(	NN	O	O
134	NN	O	O
base	NN	O	O
pair	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
premature	NN	O	O
termination	NN	O	O
codon	NN	O	O
.	NN	O	O

In	NN	O	O
studies	NN	O	O
of	NN	O	O
eight	NN	O	O
kindred	NN	O	O
,	NN	O	O
the	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
C2Q0	NN	O	O
alleles	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
common	NN	O	O
type	NN	O	O
I	NN	O	O
deficient	NN	O	O
complotype	NN	O	O
/	NN	O	O
haplotype	NN	O	O
;	NN	O	O
this	NN	O	O
deletion	NN	O	O
was	NN	O	O
not	NN	O	O
present	NN	O	O
in	NN	O	O
normal	NN	O	O
C2	NN	O	O
nor	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
C2	NN	O	I-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
genes	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
1	NN	O	O
)	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
human	NN	O	I-Disease
complement	NN	O	I-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
genomic	NN	O	O
deletion	NN	O	O
that	NN	O	O
causes	NN	O	O
skipping	NN	O	O
of	NN	O	O
exon	NN	O	O
6	NN	O	O
during	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
premature	NN	O	O
termination	NN	O	O
codon	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
type	NN	O	O
I	NN	O	O
C2Q0	NN	O	O
gene	NN	O	O
is	NN	O	O
strongly	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
HLA	NN	O	O
haplotype	NN	O	O
/	NN	O	O
complotype	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
,	NN	O	O
C2Q0	NN	O	O
,	NN	O	O
BfS	NN	O	O
,	NN	O	O
C4A4	NN	O	O
,	NN	O	O
C4B2	NN	O	O
,	NN	O	O
Drw2	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
all	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
individuals	NN	O	O
with	NN	O	O
this	NN	O	O
haplotype	NN	O	O
/	NN	O	O
complotype	NN	O	O
will	NN	O	O
harbor	NN	O	O
the	NN	O	O
28	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
C2	NN	O	O
gene	NN	O	O
deletion	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
different	NN	O	O
,	NN	O	O
as	NN	O	O
yet	NN	O	O
uncharacterized	NN	O	O
,	NN	O	O
molecular	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
two	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
and	NN	O	O
one	NN	O	O
polymorphism	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
structure	NN	O	O
-	NN	O	O
function	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyltransferase	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
two	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
,	NN	O	O
L74P	NN	O	O
and	NN	O	O
F171S	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
polymorphism	NN	O	O
,	NN	O	O
S135L	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyltransferase	NN	O	O
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
result	NN	O	O
in	NN	O	O
reduced	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
when	NN	O	O
reconstructed	NN	O	O
in	NN	O	O
the	NN	O	O
cDNA	NN	O	O
and	NN	O	O
overexpressed	NN	O	O
.	NN	O	O

The	NN	O	O
polymorphism	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
,	NN	O	O
has	NN	O	O
near	NN	O	O
normal	NN	O	O
activity	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
affect	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
residues	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
functionally	NN	O	O
important	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
polymorphism	NN	O	O
occurs	NN	O	O
in	NN	O	O
a	NN	O	O
nonconserved	NN	O	O
domain	NN	O	O
which	NN	O	O
is	NN	O	O
presumably	NN	O	O
not	NN	O	O
critical	NN	O	O
for	NN	O	O
enzymatic	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
F171S	NN	O	O
mutation	NN	O	O
is	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	O
active	NN	O	O
-	NN	O	O
site	NN	O	O
nucleophile	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
further	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
of	NN	O	O
molecular	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
galactosemia	NN	O	B-Disease
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
and	NN	O	O
GALT	NN	O	O
polymorphisms	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
tools	NN	O	O
in	NN	O	O
highlighting	NN	O	O
different	NN	O	O
functional	NN	O	O
domains	NN	O	O
in	NN	O	O
human	NN	O	O
GALT	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
relationship	NN	O	O
of	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
HLA	NN	O	O
and	NN	O	O
glyoxalase	NN	O	O
I	NN	O	O
loci	NN	O	O
.	NN	O	O

Immunogenetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
homozygous	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
individual	NN	O	O
and	NN	O	O
family	NN	O	O
members	NN	O	O
demonstrated	NN	O	O
linkage	NN	O	O
of	NN	O	O
HLA	NN	O	O
-	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
and	NN	O	O
C2o	NN	O	O
.	NN	O	O

HLA	NN	O	O
-	NN	O	O
D	NN	O	O
typing	NN	O	O
showed	NN	O	O
that	NN	O	O
5	NN	O	O
members	NN	O	O
typed	NN	O	O
with	NN	O	O
homozygous	NN	O	O
Dw2	NN	O	O
typing	NN	O	O
cells	NN	O	O
from	NN	O	O
an	NN	O	O
individual	NN	O	O
with	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
but	NN	O	O
not	NN	O	O
with	NN	O	O
Dw2	NN	O	O
typing	NN	O	O
cells	NN	O	O
from	NN	O	O
2	NN	O	O
individuals	NN	O	O
with	NN	O	O
normal	NN	O	O
C2	NN	O	O
.	NN	O	O

The	NN	O	O
homozygous	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
propositus	NN	O	O
and	NN	O	O
brother	NN	O	O
were	NN	O	O
HLA	NN	O	O
-	NN	O	O
A	NN	O	O
and	NN	O	O
B	NN	O	O
homozygous	NN	O	O
but	NN	O	O
heterozygous	NN	O	O
at	NN	O	O
the	NN	O	O
HLA	NN	O	O
-	NN	O	O
D	NN	O	O
and	NN	O	O
glyoxalase	NN	O	O
I	NN	O	O
loci	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
the	NN	O	O
C2o	NN	O	O
gene	NN	O	O
is	NN	O	O
linked	NN	O	O
with	NN	O	O
two	NN	O	O
distinct	NN	O	O
haplotypes	NN	O	O
HLA	NN	O	O
-	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
,	NN	O	O
Dw2	NN	O	O
,	NN	O	O
GLO1	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
A25	NN	O	O
,	NN	O	O
B18	NN	O	O
,	NN	O	O
D	NN	O	O
unknown	NN	O	O
,	NN	O	O
GL02	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
could	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
an	NN	O	O
ancestral	NN	O	O
recombinant	NN	O	O
event	NN	O	O
,	NN	O	O
which	NN	O	O
occurred	NN	O	O
between	NN	O	O
the	NN	O	O
C2o	NN	O	O
locus	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
D	NN	O	O
locus	NN	O	O
in	NN	O	O
which	NN	O	O
C2o	NN	O	O
segregated	NN	O	O
with	NN	O	O
HLA	NN	O	O
-	NN	O	O
B	NN	O	O
.	NN	O	O

This	NN	O	O
would	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
locus	NN	O	O
for	NN	O	O
the	NN	O	O
C2o	NN	O	O
gene	NN	O	O
maps	NN	O	O
between	NN	O	O
HLA	NN	O	O
-	NN	O	O
B	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
D	NN	O	O
on	NN	O	O
the	NN	O	O
sixth	NN	O	O
chromosome	NN	O	O
.	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
'	NN	O	I-Disease
tumor	NN	O	I-Disease
suppressor	NN	O	O
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
abnormal	NN	O	O
urogenital	NN	O	O
development	NN	O	O
in	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
human	NN	O	O
condition	NN	O	O
in	NN	O	O
which	NN	O	O
severe	NN	O	O
urogenital	NN	O	B-Disease
aberrations	NN	O	I-Disease
result	NN	O	O
in	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
,	NN	O	O
pseudohermaphroditism	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
(	NN	O	O
nephroblastoma	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
its	NN	O	O
possible	NN	O	O
role	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
coding	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
suppressor	NN	O	O
gene	NN	O	O
(	NN	O	O
WT1	NN	O	O
)	NN	O	O
for	NN	O	O
germline	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
ten	NN	O	O
independent	NN	O	O
cases	NN	O	O
of	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
zinc	NN	O	O
finger	NN	O	O
domains	NN	O	O
of	NN	O	O
one	NN	O	O
WT1	NN	O	O
gene	NN	O	O
copy	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

Nine	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
are	NN	O	O
found	NN	O	O
within	NN	O	O
exon	NN	O	O
9	NN	O	O
(	NN	O	O
zinc	NN	O	O
finger	NN	O	O
III	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
remaining	NN	O	O
mutation	NN	O	O
is	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
(	NN	O	O
zinc	NN	O	O
finger	NN	O	O
II	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
directly	NN	O	O
affect	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
recognition	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
families	NN	O	O
analyzed	NN	O	O
,	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
arise	NN	O	O
de	NN	O	O
novo	NN	O	O
.	NN	O	O

Wilms	NN	O	B-Disease
tumors	NN	O	I-Disease
from	NN	O	O
three	NN	O	O
individuals	NN	O	O
and	NN	O	O
one	NN	O	O
juvenile	NN	O	O
granulosa	NN	O	O
cell	NN	O	O
tumor	NN	O	B-Disease
demonstrate	NN	O	O
reduction	NN	O	O
to	NN	O	O
homozygosity	NN	O	O
for	NN	O	O
the	NN	O	O
mutated	NN	O	O
WT1	NN	O	O
allele	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
direct	NN	O	O
role	NN	O	O
for	NN	O	O
WT1	NN	O	O
in	NN	O	O
Denys	NN	O	B-Disease
-	NN	O	I-Disease
Drash	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
thus	NN	O	O
urogenital	NN	O	O
system	NN	O	O
development	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
and	NN	O	O
application	NN	O	O
in	NN	O	O
post	NN	O	O
-	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
performed	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	O
markers	NN	O	O
DXS52	NN	O	O
and	NN	O	O
the	NN	O	O
clotting	NN	O	O
factor	NN	O	O
VIII	NN	O	O
gene	NN	O	O
(	NN	O	O
F8C	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
several	NN	O	O
large	NN	O	O
families	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
tight	NN	O	O
linkage	NN	O	O
to	NN	O	O
DXS52	NN	O	O
could	NN	O	O
be	NN	O	O
extended	NN	O	O
giving	NN	O	O
a	NN	O	O
maximal	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
22	NN	O	O
.	NN	O	O

5	NN	O	O
at	NN	O	O
1	NN	O	O
cM	NN	O	O
.	NN	O	O

F8C	NN	O	O
was	NN	O	O
also	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
ALD	NN	O	B-Disease
with	NN	O	O
a	NN	O	O
maximal	NN	O	O
LOD	NN	O	O
score	NN	O	O
of	NN	O	O
7	NN	O	O
.	NN	O	O

8	NN	O	O
without	NN	O	O
recombination	NN	O	O
.	NN	O	O

Multipoint	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
the	NN	O	O
markers	NN	O	O
DXS304	NN	O	O
,	NN	O	O
DXS52	NN	O	O
,	NN	O	O
and	NN	O	O
F8C	NN	O	O
indicated	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
for	NN	O	O
F8C	NN	O	O
are	NN	O	O
distal	NN	O	O
to	NN	O	O
DXS52	NN	O	O
.	NN	O	O

In	NN	O	O
four	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
,	NN	O	O
no	NN	O	O
major	NN	O	O
structural	NN	O	O
rearrangement	NN	O	O
in	NN	O	O
the	NN	O	O
Xqter	NN	O	O
region	NN	O	O
was	NN	O	O
observed	NN	O	O
;	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
abnormalities	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
vision	NN	O	I-Disease
blindness	NN	O	I-Disease
genes	NN	O	I-Disease
.	NN	O	O

DNA	NN	O	O
analysis	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
use	NN	O	O
in	NN	O	O
determination	NN	O	O
of	NN	O	O
the	NN	O	O
carrier	NN	O	O
status	NN	O	O
of	NN	O	O
females	NN	O	O
at	NN	O	O
risk	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
particular	NN	O	O
family	NN	O	O
,	NN	O	O
and	NN	O	O
for	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

Two	NN	O	O
distinct	NN	O	O
mutations	NN	O	O
at	NN	O	O
a	NN	O	O
single	NN	O	O
BamHI	NN	O	O
site	NN	O	O
in	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O

Classical	NN	O	B-Disease
phenylketonuria	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hepatic	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
abolition	NN	O	O
of	NN	O	O
an	NN	O	O
invariant	NN	O	O
BamHI	NN	O	O
site	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
(	NN	O	O
exon	NN	O	O
7	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
two	NN	O	O
new	NN	O	O
point	NN	O	O
mutations	NN	O	O
at	NN	O	O
codon	NN	O	O
272	NN	O	O
and	NN	O	O
273	NN	O	O
(	NN	O	O
272gly	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
stop	NN	O	O
and	NN	O	O
273ser	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
phe	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
north	NN	O	O
eastern	NN	O	O
France	NN	O	O
or	NN	O	O
Belgium	NN	O	O
and	NN	O	O
occurred	NN	O	O
on	NN	O	O
the	NN	O	O
background	NN	O	O
of	NN	O	O
RFLP	NN	O	O
haplotype	NN	O	O
7	NN	O	O
alleles	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
supports	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
the	NN	O	O
clinical	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
PKU	NN	O	B-Disease
is	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
the	NN	O	O
large	NN	O	O
variety	NN	O	O
of	NN	O	O
mutant	NN	O	O
genotypes	NN	O	O
associated	NN	O	O
with	NN	O	O
PAH	NN	O	B-Disease
deficiencies	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
detailed	NN	O	O
multipoint	NN	O	O
map	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
4	NN	O	O
provides	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
heterogeneity	NN	O	O
and	NN	O	O
position	NN	O	O
-	NN	O	O
specific	NN	O	O
recombination	NN	O	O
rates	NN	O	O
.	NN	O	O

Utilizing	NN	O	O
the	NN	O	O
CEPH	NN	O	O
reference	NN	O	O
panel	NN	O	O
and	NN	O	O
genotypic	NN	O	O
data	NN	O	O
for	NN	O	O
53	NN	O	O
markers	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
constructed	NN	O	O
a	NN	O	O
20	NN	O	O
-	NN	O	O
locus	NN	O	O
multipoint	NN	O	O
genetic	NN	O	O
map	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
4	NN	O	O
.	NN	O	O

New	NN	O	O
RFLPs	NN	O	O
are	NN	O	O
reported	NN	O	O
for	NN	O	O
four	NN	O	O
loci	NN	O	O
.	NN	O	O

The	NN	O	O
map	NN	O	O
integrates	NN	O	O
a	NN	O	O
high	NN	O	O
-	NN	O	O
resolution	NN	O	O
genetic	NN	O	O
map	NN	O	O
of	NN	O	O
4p16	NN	O	O
into	NN	O	O
a	NN	O	O
continuous	NN	O	O
map	NN	O	O
extending	NN	O	O
to	NN	O	O
4q31	NN	O	O
and	NN	O	O
an	NN	O	O
unlinked	NN	O	O
cluster	NN	O	O
of	NN	O	O
three	NN	O	O
loci	NN	O	O
at	NN	O	O
4q35	NN	O	O
.	NN	O	O

The	NN	O	O
20	NN	O	O
linked	NN	O	O
markers	NN	O	O
form	NN	O	O
a	NN	O	O
continuous	NN	O	O
linkage	NN	O	O
group	NN	O	O
of	NN	O	O
152	NN	O	O
cM	NN	O	O
in	NN	O	O
males	NN	O	O
and	NN	O	O
202	NN	O	O
cM	NN	O	O
in	NN	O	O
females	NN	O	O
.	NN	O	O

Likely	NN	O	O
genetic	NN	O	O
locations	NN	O	O
are	NN	O	O
provided	NN	O	O
for	NN	O	O
25	NN	O	O
polymorphic	NN	O	O
anonymous	NN	O	O
sequences	NN	O	O
and	NN	O	O
28	NN	O	O
gene	NN	O	O
-	NN	O	O
specific	NN	O	O
RFLPs	NN	O	O
.	NN	O	O

The	NN	O	O
map	NN	O	O
was	NN	O	O
constructed	NN	O	O
employing	NN	O	O
the	NN	O	O
LINKAGE	NN	O	O
and	NN	O	O
CRIMAP	NN	O	O
computational	NN	O	O
methodologies	NN	O	O
to	NN	O	O
build	NN	O	O
the	NN	O	O
multipoint	NN	O	O
map	NN	O	O
via	NN	O	O
a	NN	O	O
stepwise	NN	O	O
algorithm	NN	O	O
.	NN	O	O

A	NN	O	O
detailed	NN	O	O
10	NN	O	O
-	NN	O	O
point	NN	O	O
map	NN	O	O
of	NN	O	O
the	NN	O	O
4p16	NN	O	O
region	NN	O	O
constructed	NN	O	O
from	NN	O	O
the	NN	O	O
CEPH	NN	O	O
panel	NN	O	O
provides	NN	O	O
evidence	NN	O	O
for	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
the	NN	O	O
linkage	NN	O	O
maps	NN	O	O
constructed	NN	O	O
from	NN	O	O
families	NN	O	O
segregating	NN	O	O
for	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

It	NN	O	O
additionally	NN	O	O
provides	NN	O	O
evidence	NN	O	O
for	NN	O	O
position	NN	O	O
-	NN	O	O
specific	NN	O	O
recombination	NN	O	O
frequencies	NN	O	O
in	NN	O	O
the	NN	O	O
telomeric	NN	O	O
region	NN	O	O
of	NN	O	O
4p	NN	O	O
.	NN	O	O
.	NN	O	O

Carrier	NN	O	O
detection	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
anonymous	NN	O	O
DNA	NN	O	O
polymorphisms	NN	O	O
and	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
cDNA	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
carrier	NN	O	O
identification	NN	O	O
and	NN	O	O
a	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
using	NN	O	O
DNA	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
2	NN	O	O
families	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Pelizaeus	NN	O	I-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
PMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
families	NN	O	O
,	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
single	NN	O	O
affected	NN	O	O
male	NN	O	O
could	NN	O	O
be	NN	O	O
traced	NN	O	O
back	NN	O	O
to	NN	O	O
his	NN	O	O
unaffected	NN	O	O
maternal	NN	O	O
grandfather	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
each	NN	O	O
family	NN	O	O
contains	NN	O	O
a	NN	O	O
new	NN	O	O
mutation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
the	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
the	NN	O	O
fetus	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
cytogenetic	NN	O	O
analysis	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
female	NN	O	O
,	NN	O	O
who	NN	O	O
we	NN	O	O
predict	NN	O	O
will	NN	O	O
be	NN	O	O
a	NN	O	O
noncarrier	NN	O	O
of	NN	O	O
PMD	NN	O	B-Disease
based	NN	O	O
on	NN	O	O
her	NN	O	O
genotype	NN	O	O
with	NN	O	O
the	NN	O	O
PLP	NN	O	O
intragenic	NN	O	O
polymorphism	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
deletion	NN	O	O
mutations	NN	O	O
and	NN	O	O
three	NN	O	O
new	NN	O	O
genes	NN	O	O
at	NN	O	O
the	NN	O	O
familial	NN	O	B-Disease
polyposis	NN	O	I-Disease
locus	NN	O	O
.	NN	O	O

Small	NN	O	O
(	NN	O	O
100	NN	O	O
-	NN	O	O
260	NN	O	O
kb	NN	O	O
)	NN	O	O
,	NN	O	O
nested	NN	O	O
deletions	NN	O	O
were	NN	O	O
characterized	NN	O	O
in	NN	O	O
DNA	NN	O	O
from	NN	O	O
two	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Three	NN	O	O
candidate	NN	O	O
genes	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
deleted	NN	O	O
region	NN	O	O
were	NN	O	O
ascertained	NN	O	O
and	NN	O	O
a	NN	O	O
previous	NN	O	O
candidate	NN	O	O
gene	NN	O	O
,	NN	O	O
MCC	NN	O	O
,	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
located	NN	O	O
outside	NN	O	O
the	NN	O	O
deleted	NN	O	O
region	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
new	NN	O	O
genes	NN	O	O
contained	NN	O	O
sequence	NN	O	O
identical	NN	O	O
to	NN	O	O
SRP19	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
19	NN	O	O
kd	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
ribosomal	NN	O	O
signal	NN	O	O
recognition	NN	O	O
particle	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
,	NN	O	O
provisionally	NN	O	O
designated	NN	O	O
DP1	NN	O	O
(	NN	O	O
deleted	NN	O	O
in	NN	O	O
polyposis	NN	O	O
1	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
transcribed	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
orientation	NN	O	O
as	NN	O	O
MCC	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
cDNAs	NN	O	O
,	NN	O	O
DP2	NN	O	O
and	NN	O	O
DP3	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
overlap	NN	O	O
,	NN	O	O
forming	NN	O	O
a	NN	O	O
single	NN	O	O
gene	NN	O	O
,	NN	O	O
DP2	NN	O	O
.	NN	O	O

5	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
transcribed	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
orientation	NN	O	O
as	NN	O	O
SRP19	NN	O	O
.	NN	O	O

Exclusion	NN	O	O
of	NN	O	O
linkage	NN	O	O
between	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
human	NN	O	O
serum	NN	O	O
amyloid	NN	O	O
A	NN	O	O
(	NN	O	O
SAA	NN	O	O
)	NN	O	O
gene	NN	O	O
cluster	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
trait	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
the	NN	O	O
serum	NN	O	O
amyloid	NN	O	O
A	NN	O	O
(	NN	O	O
SAA	NN	O	O
)	NN	O	O
genes	NN	O	O
by	NN	O	O
comparing	NN	O	O
alleles	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
polymorphic	NN	O	O
dinucleotide	NN	O	O
repeat	NN	O	O
and	NN	O	O
a	NN	O	O
conventional	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
RFLP	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
SAA	NN	O	O
gene	NN	O	O
cluster	NN	O	O
in	NN	O	O
Israeli	NN	O	O
FMF	NN	O	B-Disease
kindreds	NN	O	O
.	NN	O	O

By	NN	O	O
haplotype	NN	O	O
analysis	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
a	NN	O	O
minimum	NN	O	O
crossover	NN	O	O
frequency	NN	O	O
of	NN	O	O
22	NN	O	O
%	NN	O	O
between	NN	O	O
the	NN	O	O
SAA	NN	O	O
gene	NN	O	O
marker	NN	O	O
and	NN	O	O
FMF	NN	O	B-Disease
.	NN	O	O

By	NN	O	O
conventional	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
this	NN	O	O
eliminates	NN	O	O
a	NN	O	O
minimum	NN	O	O
of	NN	O	O
10	NN	O	O
.	NN	O	O

4	NN	O	O
cM	NN	O	O
including	NN	O	O
and	NN	O	O
surrounding	NN	O	O
the	NN	O	O
SAA	NN	O	O
gene	NN	O	O
cluster	NN	O	O
as	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
FMF	NN	O	B-Disease
mutation	NN	O	O
although	NN	O	O
SAA	NN	O	O
proteins	NN	O	O
are	NN	O	O
prominent	NN	O	O
physiologic	NN	O	O
markers	NN	O	O
of	NN	O	O
the	NN	O	O
acute	NN	O	O
attacks	NN	O	O
.	NN	O	O

PRAD1	NN	O	O
,	NN	O	O
a	NN	O	O
candidate	NN	O	O
BCL1	NN	O	O
oncogene	NN	O	O
:	NN	O	O
mapping	NN	O	O
and	NN	O	O
expression	NN	O	O
in	NN	O	O
centrocytic	NN	O	B-Disease
lymphoma	NN	O	I-Disease
.	NN	O	O

Rearrangement	NN	O	O
of	NN	O	O
the	NN	O	O
BCL1	NN	O	O
(	NN	O	O
B	NN	O	O
-	NN	O	O
cell	NN	O	O
lymphoma	NN	O	O
1	NN	O	O
)	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
11q13	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
characteristic	NN	O	O
of	NN	O	O
centrocytic	NN	O	B-Disease
lymphoma	NN	O	I-Disease
and	NN	O	O
also	NN	O	O
is	NN	O	O
found	NN	O	O
infrequently	NN	O	O
in	NN	O	O
other	NN	O	O
B	NN	O	B-Disease
-	NN	O	I-Disease
cell	NN	O	I-Disease
neoplasms	NN	O	I-Disease
.	NN	O	O

Rearrangement	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
deregulate	NN	O	O
a	NN	O	O
nearby	NN	O	O
protooncogene	NN	O	O
,	NN	O	O
but	NN	O	O
transcribed	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
immediate	NN	O	O
vicinity	NN	O	O
of	NN	O	O
BCL1	NN	O	O
breakpoints	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

PRAD1	NN	O	O
,	NN	O	O
previously	NN	O	O
designated	NN	O	O
D11S287E	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
on	NN	O	O
11q13	NN	O	O
as	NN	O	O
a	NN	O	O
chromosomal	NN	O	O
breakpoint	NN	O	O
region	NN	O	O
rearranged	NN	O	O
with	NN	O	O
the	NN	O	O
parathyroid	NN	O	O
hormone	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
parathyroid	NN	O	B-Disease
adenomas	NN	O	I-Disease
;	NN	O	O
this	NN	O	O
highly	NN	O	O
conserved	NN	O	O
putative	NN	O	O
oncogene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
novel	NN	O	O
cyclin	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
BCL1	NN	O	O
and	NN	O	O
implicated	NN	O	O
also	NN	O	O
in	NN	O	O
subsets	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
squamous	NN	O	I-Disease
cell	NN	O	I-Disease
neoplasms	NN	O	I-Disease
with	NN	O	O
11q13	NN	O	O
amplification	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
pulsed	NN	O	O
-	NN	O	O
field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
data	NN	O	O
showing	NN	O	O
BCL1	NN	O	O
and	NN	O	O
PRAD1	NN	O	O
to	NN	O	O
be	NN	O	O
no	NN	O	O
more	NN	O	O
than	NN	O	O
130	NN	O	O
kilobases	NN	O	O
apart	NN	O	O
.	NN	O	O

PRAD1	NN	O	O
mRNA	NN	O	O
is	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
seven	NN	O	O
of	NN	O	O
seven	NN	O	O
centrocytic	NN	O	B-Disease
lymphomas	NN	O	I-Disease
(	NN	O	O
Kiel	NN	O	O
classification	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
13	NN	O	O
closely	NN	O	O
related	NN	O	O
but	NN	O	O
noncentrocytic	NN	O	B-Disease
lymphomas	NN	O	I-Disease
.	NN	O	O

Three	NN	O	O
of	NN	O	O
the	NN	O	O
seven	NN	O	O
centrocytic	NN	O	B-Disease
lymphomas	NN	O	I-Disease
had	NN	O	O
detectable	NN	O	O
BCL1	NN	O	O
DNA	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
two	NN	O	O
unusual	NN	O	O
cases	NN	O	O
of	NN	O	O
CLL	NN	O	O
with	NN	O	O
BCL1	NN	O	O
rearrangement	NN	O	O
overexpressed	NN	O	O
PRAD1	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
five	NN	O	O
CLL	NN	O	O
controls	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
PRAD1	NN	O	O
is	NN	O	O
an	NN	O	O
excellent	NN	O	O
candidate	NN	O	O
"	NN	O	O
BCL1	NN	O	O
oncogene	NN	O	O
.	NN	O	O

"	NN	O	O
Its	NN	O	O
overexpression	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
key	NN	O	O
consequence	NN	O	O
of	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
the	NN	O	O
BCL1	NN	O	O
vicinity	NN	O	O
in	NN	O	O
B	NN	O	B-Disease
-	NN	O	I-Disease
cell	NN	O	I-Disease
neoplasms	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
unifying	NN	O	O
pathogenetic	NN	O	O
feature	NN	O	O
in	NN	O	O
centrocytic	NN	O	B-Disease
lymphoma	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
new	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
juvenile	NN	O	O
metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
patient	NN	O	O
.	NN	O	O

Fragments	NN	O	O
of	NN	O	O
the	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
gene	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
juvenile	NN	O	O
-	NN	O	O
onset	NN	O	O
metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
were	NN	O	O
amplified	NN	O	O
by	NN	O	O
PCR	NN	O	O
and	NN	O	O
ligated	NN	O	O
into	NN	O	O
MP13	NN	O	O
cloning	NN	O	O
vectors	NN	O	O
.	NN	O	O

Clones	NN	O	O
hybridizing	NN	O	O
with	NN	O	O
cDNA	NN	O	O
for	NN	O	O
human	NN	O	O
ARSA	NN	O	O
were	NN	O	O
selected	NN	O	O
,	NN	O	O
examined	NN	O	O
for	NN	O	O
appropriate	NN	O	O
size	NN	O	O
inserts	NN	O	O
,	NN	O	O
and	NN	O	O
used	NN	O	O
to	NN	O	O
prepare	NN	O	O
single	NN	O	O
-	NN	O	O
stranded	NN	O	O
phage	NN	O	O
DNA	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
and	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
intronic	NN	O	O
sequence	NN	O	O
revealed	NN	O	O
two	NN	O	O
putative	NN	O	O
disease	NN	O	O
-	NN	O	O
related	NN	O	O
mutations	NN	O	O
.	NN	O	O

One	NN	O	O
,	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
isoleucine	NN	O	O
by	NN	O	O
serine	NN	O	O
.	NN	O	O

Introduction	NN	O	O
of	NN	O	O
this	NN	O	O
alteration	NN	O	O
into	NN	O	O
the	NN	O	O
normal	NN	O	O
ARSA	NN	O	O
cDNA	NN	O	O
sequence	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
substantial	NN	O	O
decrease	NN	O	O
in	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
on	NN	O	O
transient	NN	O	O
expression	NN	O	O
in	NN	O	O
cultured	NN	O	O
baby	NN	O	O
hamster	NN	O	O
kidney	NN	O	O
cells	NN	O	O
.	NN	O	O

About	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
small	NN	O	O
residual	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
mutated	NN	O	O
ARSA	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
,	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
allele	NN	O	O
and	NN	O	O
resulted	NN	O	O
in	NN	O	O
an	NN	O	O
altered	NN	O	O
splice	NN	O	O
-	NN	O	O
recognition	NN	O	O
sequence	NN	O	O
between	NN	O	O
exon	NN	O	O
7	NN	O	O
and	NN	O	O
the	NN	O	O
following	NN	O	O
intron	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
a	NN	O	O
restriction	NN	O	O
site	NN	O	O
.	NN	O	O

Apparently	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
mRNA	NN	O	O
were	NN	O	O
generated	NN	O	O
from	NN	O	O
this	NN	O	O
allele	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
or	NN	O	O
immuno	NN	O	O
-	NN	O	O
cross	NN	O	O
-	NN	O	O
reactive	NN	O	O
material	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

A	NN	O	O
collection	NN	O	O
of	NN	O	O
DNA	NN	O	O
samples	NN	O	O
from	NN	O	O
known	NN	O	O
or	NN	O	O
suspected	NN	O	O
MLD	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
their	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
was	NN	O	O
screened	NN	O	O
for	NN	O	O
these	NN	O	O
mutations	NN	O	O
.	NN	O	O

Four	NN	O	O
additional	NN	O	O
individuals	NN	O	O
carrying	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
among	NN	O	O
the	NN	O	O
nearly	NN	O	O
100	NN	O	O
MLD	NN	O	B-Disease
patients	NN	O	O
in	NN	O	O
the	NN	O	O
sample	NN	O	O
.	NN	O	O

Gene	NN	O	O
segregation	NN	O	O
in	NN	O	O
the	NN	O	O
original	NN	O	O
patients	NN	O	O
family	NN	O	O
was	NN	O	O
consistent	NN	O	O
with	NN	O	O
available	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
data	NN	O	O
.	NN	O	O

No	NN	O	O
individuals	NN	O	O
homozygous	NN	O	O
for	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
combinations	NN	O	O
with	NN	O	O
other	NN	O	O
MLD	NN	O	B-Disease
mutations	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
does	NN	O	O
result	NN	O	O
in	NN	O	O
some	NN	O	O
residual	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
late	NN	O	O
-	NN	O	O
onset	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
following	NN	O	O
exon	NN	O	O
7	NN	O	O
produces	NN	O	O
late	NN	O	O
-	NN	O	O
infantile	NN	O	O
MLD	NN	O	B-Disease
when	NN	O	O
combined	NN	O	O
with	NN	O	O
other	NN	O	O
enzyme	NN	O	O
-	NN	O	O
null	NN	O	O
mutations	NN	O	O
,	NN	O	O
implying	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
completely	NN	O	O
silent	NN	O	O
enzymatically	NN	O	O
.	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
and	NN	O	O
presumptive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
in	NN	O	O
hematopoietic	NN	O	O
cell	NN	O	O
lineages	NN	O	O
of	NN	O	O
a	NN	O	O
thrombocytopenic	NN	O	O
carrier	NN	O	O
female	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
thrombocytopenic	NN	O	B-Disease
female	NN	O	O
belonging	NN	O	O
to	NN	O	O
a	NN	O	O
pedigree	NN	O	O
with	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
RFLP	NN	O	O
)	NN	O	O
analysis	NN	O	O
with	NN	O	O
probe	NN	O	O
M27	NN	O	O
beta	NN	O	O
,	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
she	NN	O	O
is	NN	O	O
a	NN	O	O
carrier	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O

Both	NN	O	O
small	NN	O	O
-	NN	O	O
sized	NN	O	O
and	NN	O	O
normal	NN	O	O
-	NN	O	O
sized	NN	O	O
platelets	NN	O	O
were	NN	O	O
present	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
carriers	NN	O	O
,	NN	O	O
she	NN	O	O
does	NN	O	O
not	NN	O	O
manifest	NN	O	O
nonrandom	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
in	NN	O	O
the	NN	O	O
thrombopoietic	NN	O	O
cell	NN	O	O
lineage	NN	O	O
.	NN	O	O

Study	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
RFLP	NN	O	O
and	NN	O	O
methylation	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
was	NN	O	O
nonrandom	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
but	NN	O	O
random	NN	O	O
in	NN	O	O
granulocytes	NN	O	O
.	NN	O	O

While	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
complete	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
thrombocytopenia	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
carrier	NN	O	O
female	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
as	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
atypical	NN	O	O
lyonization	NN	O	O
,	NN	O	O
it	NN	O	O
also	NN	O	O
suggests	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
occurs	NN	O	O
at	NN	O	O
(	NN	O	O
or	NN	O	O
extends	NN	O	O
up	NN	O	O
to	NN	O	O
)	NN	O	O
a	NN	O	O
later	NN	O	O
stage	NN	O	O
than	NN	O	O
the	NN	O	O
multipotent	NN	O	O
stem	NN	O	O
cell	NN	O	O
along	NN	O	O
the	NN	O	O
hematopoietic	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
German	NN	O	O
family	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
exon	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
2	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
two	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
in	NN	O	O
a	NN	O	O
German	NN	O	O
family	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
,	NN	O	O
which	NN	O	O
causes	NN	O	O
loss	NN	O	O
of	NN	O	O
an	NN	O	O
HphI	NN	O	O
site	NN	O	O
and	NN	O	O
changes	NN	O	O
amino	NN	O	O
acid	NN	O	O
155	NN	O	O
from	NN	O	O
threonine	NN	O	O
to	NN	O	O
isoleucine	NN	O	O
,	NN	O	O
was	NN	O	O
absent	NN	O	O
from	NN	O	O
108	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
5	NN	O	O
concordances	NN	O	O
and	NN	O	O
1	NN	O	O
discrepancy	NN	O	O
between	NN	O	O
these	NN	O	O
results	NN	O	O
and	NN	O	O
those	NN	O	O
obtained	NN	O	O
by	NN	O	O
magnetic	NN	O	O
resonance	NN	O	O
imaging	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
3	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
kinetic	NN	O	O
variant	NN	O	O
of	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
a	NN	O	I-Disease
hepatic	NN	O	I-Disease
enzyme	NN	O	I-Disease
,	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
PAH	NN	O	O
activity	NN	O	O
results	NN	O	O
in	NN	O	O
typical	NN	O	O
PKU	NN	O	B-Disease
while	NN	O	O
persistence	NN	O	O
of	NN	O	O
a	NN	O	O
residual	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
gives	NN	O	O
rise	NN	O	O
to	NN	O	O
variant	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
a	NN	O	O
3	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
(	NN	O	O
delta	NN	O	O
194	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
mild	NN	O	O
variant	NN	O	O
,	NN	O	O
with	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
for	NN	O	O
phenylalanine	NN	O	O
(	NN	O	O
Km	NN	O	O
=	NN	O	O
160	NN	O	O
nM	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
provide	NN	O	O
functional	NN	O	O
evidence	NN	O	O
for	NN	O	O
responsibility	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
mutant	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
deletion	NN	O	O
was	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
presents	NN	O	O
no	NN	O	O
homology	NN	O	O
with	NN	O	O
other	NN	O	O
hydroxylases	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
exon	NN	O	O
3	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
for	NN	O	O
phenylalanine	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
since	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
98	NN	O	O
PKU	NN	O	B-Disease
patients	NN	O	O
tested	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
this	NN	O	O
particular	NN	O	O
deletion	NN	O	O
,	NN	O	O
our	NN	O	O
study	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
molecular	NN	O	O
event	NN	O	O
probably	NN	O	O
occurred	NN	O	O
recently	NN	O	O
on	NN	O	O
the	NN	O	O
background	NN	O	O
of	NN	O	O
a	NN	O	O
haplotype	NN	O	O
2	NN	O	O
gene	NN	O	O
in	NN	O	O
Portugal	NN	O	O
.	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
KIT	NN	O	O
(	NN	O	O
mast	NN	O	O
/	NN	O	O
stem	NN	O	O
cell	NN	O	O
growth	NN	O	O
factor	NN	O	O
receptor	NN	O	O
)	NN	O	O
protooncogene	NN	O	O
in	NN	O	O
human	NN	O	O
piebaldism	NN	O	B-Disease
.	NN	O	O

Piebaldism	NN	O	B-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
cogenital	NN	O	O
patches	NN	O	O
of	NN	O	O
skin	NN	O	O
and	NN	O	O
hair	NN	O	O
from	NN	O	O
which	NN	O	O
melanocytes	NN	O	O
are	NN	O	O
completely	NN	O	O
absent	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
disorder	NN	O	O
of	NN	O	O
mouse	NN	O	O
,	NN	O	O
dominant	NN	O	O
white	NN	O	O
spotting	NN	O	O
(	NN	O	O
W	NN	O	O
)	NN	O	O
,	NN	O	O
results	NN	O	O
from	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
c	NN	O	O
-	NN	O	O
Kit	NN	O	O
protooncogene	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
and	NN	O	O
receptor	NN	O	O
for	NN	O	O
mast	NN	O	O
/	NN	O	O
stem	NN	O	O
cell	NN	O	O
growth	NN	O	O
factor	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
KIT	NN	O	O
gene	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
proband	NN	O	O
with	NN	O	O
classic	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
piebaldism	NN	O	B-Disease
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
Gly	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
Arg	NN	O	O
substitution	NN	O	O
at	NN	O	O
codon	NN	O	O
664	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
domain	NN	O	O
.	NN	O	O

This	NN	O	O
substitution	NN	O	O
was	NN	O	O
not	NN	O	O
seen	NN	O	O
in	NN	O	O
any	NN	O	O
normal	NN	O	O
individuals	NN	O	O
and	NN	O	O
was	NN	O	O
completely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
piebald	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
probands	NN	O	O
family	NN	O	O
.	NN	O	O

Piebaldism	NN	O	B-Disease
in	NN	O	O
this	NN	O	O
family	NN	O	O
thus	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
human	NN	O	O
homologue	NN	O	O
to	NN	O	O
dominant	NN	O	O
white	NN	O	O
spotting	NN	O	O
(	NN	O	O
W	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
Japanese	NN	O	O
family	NN	O	O
with	NN	O	O
normotriglyceridemic	NN	O	B-Disease
abetalipoproteinemia	NN	O	I-Disease
indicates	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
B	NN	O	O
gene	NN	O	O
.	NN	O	O

Normotriglyceridemic	NN	O	B-Disease
abetalipoproteinemia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
familial	NN	O	B-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
an	NN	O	O
isolated	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
apoB	NN	O	I-Disease
-	NN	O	I-Disease
100	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
this	NN	O	O
disease	NN	O	O
,	NN	O	O
who	NN	O	O
had	NN	O	O
normal	NN	O	O
apoB	NN	O	O
-	NN	O	O
48	NN	O	O
but	NN	O	O
no	NN	O	O
apoB	NN	O	O
-	NN	O	O
100	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
linkage	NN	O	O
of	NN	O	O
apoB	NN	O	O
gene	NN	O	O
using	NN	O	O
three	NN	O	O
genetic	NN	O	O
markers	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
and	NN	O	O
her	NN	O	O
affected	NN	O	O
brother	NN	O	O
showed	NN	O	O
completely	NN	O	O
different	NN	O	O
apoB	NN	O	O
gene	NN	O	O
alleles	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
apoB	NN	O	O
gene	NN	O	O
itself	NN	O	O
is	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
this	NN	O	O
disorder	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
an	NN	O	O
American	NN	O	O
case	NN	O	O
had	NN	O	O
a	NN	O	O
point	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
apoB	NN	O	O
gene	NN	O	O
generating	NN	O	O
an	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
this	NN	O	O
disorder	NN	O	O
can	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
defect	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
either	NN	O	O
an	NN	O	O
apoB	NN	O	O
gene	NN	O	O
or	NN	O	O
other	NN	O	O
genes	NN	O	O
.	NN	O	O
.	NN	O	O

Localisation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
to	NN	O	O
between	NN	O	O
DXS7	NN	O	O
and	NN	O	O
DXS426	NN	O	O
on	NN	O	O
Xp	NN	O	O
.	NN	O	O

A	NN	O	O
highly	NN	O	O
informative	NN	O	O
microsatellite	NN	O	O
marker	NN	O	O
,	NN	O	O
DXS426	NN	O	O
,	NN	O	O
which	NN	O	O
maps	NN	O	O
proximal	NN	O	O
to	NN	O	O
DXS7	NN	O	O
in	NN	O	O
the	NN	O	O
interval	NN	O	O
Xp11	NN	O	O
.	NN	O	O

4	NN	O	O
-	NN	O	O
Xp11	NN	O	O
4	NN	O	O
-	NN	O	O
Xp11	NN	O	O
.	NN	O	O

23	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
refine	NN	O	O
further	NN	O	O
the	NN	O	O
localisation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
NDP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
from	NN	O	O
a	NN	O	O
multiply	NN	O	O
informative	NN	O	O
crossover	NN	O	O
localize	NN	O	O
the	NN	O	O
NDP	NN	O	B-Disease
gene	NN	O	O
proximal	NN	O	O
to	NN	O	O
DXS7	NN	O	O
.	NN	O	O

In	NN	O	O
conjunction	NN	O	O
with	NN	O	O
information	NN	O	O
from	NN	O	O
2	NN	O	O
NDP	NN	O	B-Disease
patients	NN	O	O
who	NN	O	O
have	NN	O	O
a	NN	O	O
deletion	NN	O	O
for	NN	O	O
DXS7	NN	O	O
but	NN	O	O
not	NN	O	O
for	NN	O	O
DSX426	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
NDP	NN	O	B-Disease
gene	NN	O	O
lies	NN	O	O
between	NN	O	O
DXS7	NN	O	O
and	NN	O	O
DXS426	NN	O	O
on	NN	O	O
proximal	NN	O	O
Xp	NN	O	O
.	NN	O	O

Aberrant	NN	O	O
splicing	NN	O	O
of	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
mRNA	NN	O	O
:	NN	O	O
the	NN	O	O
major	NN	O	O
cause	NN	O	O
for	NN	O	O
phenylketonuria	NN	O	B-Disease
in	NN	O	O
parts	NN	O	O
of	NN	O	O
southern	NN	O	O
Europe	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
mutation	NN	O	O
within	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
gene	NN	O	O
that	NN	O	O
causes	NN	O	O
aberrant	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
that	NN	O	O
is	NN	O	O
in	NN	O	O
tight	NN	O	O
association	NN	O	O
with	NN	O	O
chromosomal	NN	O	O
haplotypes	NN	O	O
6	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
and	NN	O	O
36	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
these	NN	O	O
particular	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
Bulgaria	NN	O	O
,	NN	O	O
Italy	NN	O	O
,	NN	O	O
and	NN	O	O
Turkey	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
more	NN	O	O
frequent	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
causing	NN	O	O
classical	NN	O	O
phenylketonuria	NN	O	B-Disease
in	NN	O	O
this	NN	O	O
part	NN	O	O
of	NN	O	O
Europe	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
position	NN	O	O
546	NN	O	O
in	NN	O	O
intron	NN	O	O
10	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
,	NN	O	O
11	NN	O	O
bp	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
intron	NN	O	O
10	NN	O	O
/	NN	O	O
exon	NN	O	O
11	NN	O	O
boundary	NN	O	O
.	NN	O	O

It	NN	O	O
activates	NN	O	O
a	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
site	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
insertion	NN	O	O
of	NN	O	O
9	NN	O	O
nucleotides	NN	O	O
between	NN	O	O
exon	NN	O	O
10	NN	O	O
and	NN	O	O
exon	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
processed	NN	O	O
mRNA	NN	O	O
.	NN	O	O

Normal	NN	O	O
amounts	NN	O	O
of	NN	O	O
liver	NN	O	O
PAH	NN	O	O
protein	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
homozygous	NN	O	O
patients	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
catalytic	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

This	NN	O	O
loss	NN	O	O
of	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
is	NN	O	O
probably	NN	O	O
caused	NN	O	O
by	NN	O	O
conformational	NN	O	O
changes	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
insertion	NN	O	O
of	NN	O	O
three	NN	O	O
additional	NN	O	O
amino	NN	O	O
acids	NN	O	O
(	NN	O	O
Gly	NN	O	O
-	NN	O	O
Leu	NN	O	O
-	NN	O	O
Gln	NN	O	O
)	NN	O	O
between	NN	O	O
the	NN	O	O
normal	NN	O	O
sequences	NN	O	O
encoded	NN	O	O
by	NN	O	O
exon	NN	O	O
10	NN	O	O
and	NN	O	O
exon	NN	O	O
11	NN	O	O
.	NN	O	O
.	NN	O	O

Gardner	NN	O	B-Disease
syndrome	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
.	NN	O	O

We	NN	O	O
described	NN	O	O
a	NN	O	O
15	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
boy	NN	O	O
with	NN	O	O
Gardner	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
GS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
craniofacial	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

High	NN	O	O
-	NN	O	O
resolution	NN	O	O
banding	NN	O	O
analysis	NN	O	O
showed	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
(	NN	O	O
q22	NN	O	O
.	NN	O	O
1	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q31	NN	O	O
1	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q31	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
case	NN	O	O
and	NN	O	O
in	NN	O	O
3	NN	O	O
previously	NN	O	O
reported	NN	O	O
5q	NN	O	O
-	NN	O	O
patients	NN	O	O
with	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
GS	NN	O	B-Disease
/	NN	O	O
or	NN	O	O
familial	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
FPC	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
in	NN	O	O
the	NN	O	O
5q22	NN	O	O
region	NN	O	O
,	NN	O	O
a	NN	O	O
result	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
findings	NN	O	O
of	NN	O	O
linkage	NN	O	O
studies	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
childhood	NN	O	O
-	NN	O	O
onset	NN	O	O
Tourette	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
40	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
man	NN	O	O
with	NN	O	O
childhood	NN	O	O
-	NN	O	O
onset	NN	O	O
Tourette	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
TS	NN	O	B-Disease
)	NN	O	O
developed	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
believe	NN	O	O
this	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
first	NN	O	O
reported	NN	O	O
case	NN	O	O
of	NN	O	O
childhood	NN	O	O
-	NN	O	O
onset	NN	O	O
TS	NN	O	B-Disease
with	NN	O	O
adult	NN	O	O
onset	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

Discovery	NN	O	O
of	NN	O	O
other	NN	O	O
cases	NN	O	O
with	NN	O	O
both	NN	O	O
disorders	NN	O	O
may	NN	O	O
provide	NN	O	O
clues	NN	O	O
to	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
both	NN	O	O
conditions	NN	O	O
.	NN	O	O
.	NN	O	O

Sequence	NN	O	O
of	NN	O	O
DNA	NN	O	O
flanking	NN	O	O
the	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
rapid	NN	O	O
identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
causing	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
requires	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
readily	NN	O	O
screen	NN	O	O
the	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
most	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
mutation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
sequenced	NN	O	O
the	NN	O	O
portions	NN	O	O
of	NN	O	O
the	NN	O	O
introns	NN	O	O
flanking	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
14	NN	O	O
HEXA	NN	O	O
exons	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
specify	NN	O	O
oligonucleotide	NN	O	O
primers	NN	O	O
for	NN	O	O
the	NN	O	O
PCR	NN	O	O
-	NN	O	O
dependent	NN	O	O
amplification	NN	O	O
of	NN	O	O
each	NN	O	O
exon	NN	O	O
and	NN	O	O
splice	NN	O	O
-	NN	O	O
junction	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
amplified	NN	O	O
products	NN	O	O
were	NN	O	O
analyzed	NN	O	O
,	NN	O	O
by	NN	O	O
electrophoresis	NN	O	O
in	NN	O	O
nondenaturing	NN	O	O
polyacrylamide	NN	O	O
gels	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
either	NN	O	O
heteroduplexes	NN	O	O
,	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
annealing	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
mutant	NN	O	O
DNA	NN	O	O
strands	NN	O	O
,	NN	O	O
or	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
,	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
renaturation	NN	O	O
of	NN	O	O
single	NN	O	O
-	NN	O	O
stranded	NN	O	O
DNA	NN	O	O
.	NN	O	O

Five	NN	O	O
novel	NN	O	O
mutations	NN	O	O
from	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
patients	NN	O	O
were	NN	O	O
detected	NN	O	O
a	NN	O	O
5	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
of	NN	O	O
TCTCC	NN	O	O
in	NN	O	O
IVS	NN	O	O
-	NN	O	O
9	NN	O	O
;	NN	O	O
a	NN	O	O
2	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
of	NN	O	O
TG	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
;	NN	O	O
G78	NN	O	O
to	NN	O	O
A	NN	O	O
,	NN	O	O
giving	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
in	NN	O	O
exon	NN	O	O
1	NN	O	O
;	NN	O	O
G533	NN	O	O
to	NN	O	O
T	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
,	NN	O	O
producing	NN	O	O
the	NN	O	O
third	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
detected	NN	O	O
at	NN	O	O
this	NN	O	O
site	NN	O	O
;	NN	O	O
and	NN	O	O
G	NN	O	O
to	NN	O	O
C	NN	O	O
at	NN	O	O
position	NN	O	O
1	NN	O	O
of	NN	O	O
IVS	NN	O	O
-	NN	O	O
2	NN	O	O
,	NN	O	O
expected	NN	O	O
to	NN	O	O
produce	NN	O	O
abnormal	NN	O	O
splicing	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
two	NN	O	O
mutations	NN	O	O
,	NN	O	O
(	NN	O	O
G1496	NN	O	O
to	NN	O	O
A	NN	O	O
in	NN	O	O
exon	NN	O	O
13	NN	O	O
and	NN	O	O
a	NN	O	O
4	NN	O	O
-	NN	O	O
bp	NN	O	O
insertion	NN	O	O
in	NN	O	O
exon	NN	O	O
11	NN	O	O
)	NN	O	O
that	NN	O	O
have	NN	O	O
previously	NN	O	O
been	NN	O	O
reported	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
two	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
:	NN	O	O
correlation	NN	O	O
of	NN	O	O
mutations	NN	O	O
with	NN	O	O
highly	NN	O	O
conserved	NN	O	O
domains	NN	O	O
in	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyl	NN	O	O
transferase	NN	O	O
.	NN	O	O

Galactosemia	NN	O	B-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	I-Disease
human	NN	O	I-Disease
galactose	NN	O	I-Disease
metabolism	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
enzyme	NN	O	I-Disease
galactose	NN	O	I-Disease
-	NN	O	I-Disease
1	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
uridyl	NN	O	I-Disease
transferase	NN	O	I-Disease
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
this	NN	O	O
disorder	NN	O	O
is	NN	O	O
at	NN	O	O
present	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
which	NN	O	O
result	NN	O	O
in	NN	O	O
low	NN	O	O
or	NN	O	O
undetectable	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
591	NN	O	O
a	NN	O	O
transition	NN	O	O
which	NN	O	O
substitutes	NN	O	O
glutamine	NN	O	O
188	NN	O	O
by	NN	O	O
arginine	NN	O	O
.	NN	O	O

The	NN	O	O
mutated	NN	O	O
glutamine	NN	O	O
is	NN	O	O
not	NN	O	O
only	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
evolution	NN	O	O
(	NN	O	O
conserved	NN	O	O
also	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
and	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
also	NN	O	O
two	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
active	NN	O	O
site	NN	O	O
histidine	NN	O	O
-	NN	O	O
proline	NN	O	O
-	NN	O	O
histidine	NN	O	O
triad	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
about	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
arginine	NN	O	O
188	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
galactosemia	NN	O	B-Disease
mutation	NN	O	O
characterized	NN	O	O
to	NN	O	O
date	NN	O	O
.	NN	O	O

It	NN	O	O
accounts	NN	O	O
for	NN	O	O
one	NN	O	O
-	NN	O	O
fourth	NN	O	O
of	NN	O	O
the	NN	O	O
galactosemia	NN	O	B-Disease
alleles	NN	O	O
studied	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
arginine	NN	O	O
333	NN	O	O
by	NN	O	O
tryptophan	NN	O	O
,	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1025	NN	O	O
.	NN	O	O

The	NN	O	O
area	NN	O	O
surrounding	NN	O	O
this	NN	O	O
missense	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
highly	NN	O	O
conserved	NN	O	O
domain	NN	O	O
in	NN	O	O
the	NN	O	O
homologous	NN	O	O
enzymes	NN	O	O
from	NN	O	O
E	NN	O	O
.	NN	O	O
coli	NN	O	O
,	NN	O	O
yeast	NN	O	O
,	NN	O	O
and	NN	O	O
humans	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
mutation	NN	O	O
results	NN	O	O
in	NN	O	O
undetectable	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
is	NN	O	O
a	NN	O	O
severe	NN	O	O
mutation	NN	O	O
.	NN	O	O

This	NN	O	O
second	NN	O	O
mutation	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
rare	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
we	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
galactosemia	NN	O	B-Disease
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
,	NN	O	O
heterogeneity	NN	O	O
which	NN	O	O
might	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
variable	NN	O	O
clinical	NN	O	O
outcome	NN	O	O
observed	NN	O	O
in	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O
.	NN	O	O

Hypoxanthine	NN	O	B-Disease
-	NN	O	I-Disease
guanine	NN	O	I-Disease
phosphoribosyltransferase	NN	O	I-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
analysis	NN	O	O
of	NN	O	O
HPRT	NN	O	O
mutations	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
and	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
amplification	NN	O	O
.	NN	O	O

The	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
severe	NN	O	O
X	NN	O	B-Disease
chromosome	NN	O	I-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
human	NN	O	I-Disease
disease	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
virtual	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
hypoxanthine	NN	O	I-Disease
-	NN	O	I-Disease
guanine	NN	O	I-Disease
phosphoribosyltransferase	NN	O	I-Disease
(	NN	O	I-Disease
HPRT	NN	O	I-Disease
)	NN	O	I-Disease
activity	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
partial	NN	O	O
deficiency	NN	O	O
in	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
gouty	NN	O	B-Disease
arthritis	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
genetic	NN	O	O
basis	NN	O	O
for	NN	O	O
reduction	NN	O	O
or	NN	O	O
loss	NN	O	O
of	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
amplified	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
HPRT	NN	O	O
cDNA	NN	O	O
from	NN	O	O
four	NN	O	O
patients	NN	O	O
one	NN	O	O
with	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
HPRTPerth	NN	O	O
)	NN	O	O
and	NN	O	O
three	NN	O	O
with	NN	O	O
partial	NN	O	B-Disease
deficiencies	NN	O	I-Disease
of	NN	O	I-Disease
HPRT	NN	O	I-Disease
activity	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
designated	NN	O	O
HPRTUrangan	NN	O	O
,	NN	O	O
HPRTSwan	NN	O	O
and	NN	O	O
HPRTToowong	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
four	NN	O	O
patients	NN	O	O
,	NN	O	O
the	NN	O	O
only	NN	O	O
mutation	NN	O	O
identified	NN	O	O
was	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
substitution	NN	O	O
in	NN	O	O
exons	NN	O	O
2	NN	O	O
or	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
predicts	NN	O	O
a	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
translated	NN	O	O
protein	NN	O	O
.	NN	O	O

Each	NN	O	O
base	NN	O	O
change	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
amplification	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
interesting	NN	O	O
to	NN	O	O
note	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
found	NN	O	O
for	NN	O	O
HPRTPerth	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
reported	NN	O	O
for	NN	O	O
HPRTFlint	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
mutations	NN	O	O
are	NN	O	O
de	NN	O	O
novo	NN	O	O
events	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
and	NN	O	O
metabolic	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
metabolic	NN	O	B-Disease
disorder	NN	O	I-Disease
normotriglyceridemic	NN	O	B-Disease
abetalipoproteinemia	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
described	NN	O	O
a	NN	O	O
disorder	NN	O	O
,	NN	O	O
normotriglyceridemic	NN	O	B-Disease
abetalipoproteinemia	NN	O	I-Disease
,	NN	O	O
that	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
virtual	NN	O	O
absence	NN	O	O
of	NN	O	O
plasma	NN	O	O
low	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
and	NN	O	O
complete	NN	O	O
absence	NN	O	O
of	NN	O	O
apoB	NN	O	O
-	NN	O	O
100	NN	O	O
,	NN	O	O
but	NN	O	O
with	NN	O	O
apparently	NN	O	O
normal	NN	O	O
secretion	NN	O	O
of	NN	O	O
triglyceride	NN	O	O
-	NN	O	O
rich	NN	O	O
lipoproteins	NN	O	O
containing	NN	O	O
apoB	NN	O	O
-	NN	O	O
48	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
plasma	NN	O	O
lipoproteins	NN	O	O
were	NN	O	O
shown	NN	O	O
on	NN	O	O
polyacrylamide	NN	O	O
gels	NN	O	O
and	NN	O	O
by	NN	O	O
antibody	NN	O	O
mapping	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
new	NN	O	O
truncated	NN	O	O
apoB	NN	O	O
variant	NN	O	O
,	NN	O	O
apoB	NN	O	O
-	NN	O	O
50	NN	O	O
,	NN	O	O
circulating	NN	O	O
along	NN	O	O
with	NN	O	O
her	NN	O	O
apoB	NN	O	O
-	NN	O	O
48	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
this	NN	O	O
individual	NN	O	O
to	NN	O	O
be	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
single	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
nucleotide	NN	O	O
substitution	NN	O	O
at	NN	O	O
apoB	NN	O	O
codon	NN	O	O
2252	NN	O	O
,	NN	O	O
which	NN	O	O
produces	NN	O	O
a	NN	O	O
premature	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
example	NN	O	O
of	NN	O	O
homozygous	NN	O	B-Disease
hypobetalipoproteinemia	NN	O	I-Disease
.	NN	O	O

Electron	NN	O	O
photomicrographs	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
diameters	NN	O	O
of	NN	O	O
particles	NN	O	O
in	NN	O	O
the	NN	O	O
d	NN	O	O
less	NN	O	O
than	NN	O	O
1	NN	O	O
.	NN	O	O

006	NN	O	O
g	NN	O	O
/	NN	O	O
ml	NN	O	O
lipoprotein	NN	O	O
fraction	NN	O	O
,	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
postprandial	NN	O	O
and	NN	O	O
postabsorptive	NN	O	O
state	NN	O	O
,	NN	O	O
are	NN	O	O
bimodally	NN	O	O
distributed	NN	O	O
.	NN	O	O

The	NN	O	O
molar	NN	O	O
ratio	NN	O	O
of	NN	O	O
apoE	NN	O	O
to	NN	O	O
apoB	NN	O	O
in	NN	O	O
these	NN	O	O
particles	NN	O	O
is	NN	O	O
3	NN	O	O
.	NN	O	O

5	NN	O	O
1	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
normal	NN	O	O
VLDL	NN	O	O
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
LDL	NN	O	O
interval	NN	O	O
contains	NN	O	O
both	NN	O	O
spherical	NN	O	O
and	NN	O	O
cuboidal	NN	O	O
particles	NN	O	O
.	NN	O	O

Autologous	NN	O	O
reinfusion	NN	O	O
of	NN	O	O
labeled	NN	O	O
d	NN	O	O
less	NN	O	O
than	NN	O	O
1	NN	O	O
.	NN	O	O

006	NN	O	O
g	NN	O	O
/	NN	O	O
ml	NN	O	O
lipoproteins	NN	O	O
showed	NN	O	O
exponential	NN	O	O
disappearance	NN	O	O
from	NN	O	O
plasma	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
half	NN	O	O
-	NN	O	O
removal	NN	O	O
time	NN	O	O
of	NN	O	O
50	NN	O	O
min	NN	O	O
,	NN	O	O
somewhat	NN	O	O
slower	NN	O	O
than	NN	O	O
for	NN	O	O
normal	NN	O	O
chylomicrons	NN	O	O
but	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
for	NN	O	O
VLDL	NN	O	O
.	NN	O	O

The	NN	O	O
calculated	NN	O	O
production	NN	O	O
rate	NN	O	O
for	NN	O	O
apoB	NN	O	O
was	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
in	NN	O	O
this	NN	O	O
subject	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
origin	NN	O	O
of	NN	O	O
phenylketonuria	NN	O	B-Disease
in	NN	O	O
Yemenite	NN	O	O
Jews	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
metabolic	NN	O	B-Disease
disease	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
recessive	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
hepatic	NN	O	O
enzyme	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
incidence	NN	O	O
of	NN	O	O
PKU	NN	O	B-Disease
varies	NN	O	O
widely	NN	O	O
across	NN	O	O
different	NN	O	O
geographic	NN	O	O
areas	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
highest	NN	O	O
(	NN	O	O
about	NN	O	O
1	NN	O	O
in	NN	O	O
5	NN	O	O
,	NN	O	O
000	NN	O	O
live	NN	O	O
births	NN	O	O
)	NN	O	O
in	NN	O	O
Ireland	NN	O	O
and	NN	O	O
western	NN	O	O
Scotland	NN	O	O
,	NN	O	O
and	NN	O	O
among	NN	O	O
Yemenite	NN	O	O
Jews	NN	O	O
.	NN	O	O

A	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
account	NN	O	O
for	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
PKU	NN	O	B-Disease
cases	NN	O	O
in	NN	O	O
the	NN	O	O
European	NN	O	O
population	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
molecular	NN	O	O
defect	NN	O	O
-	NN	O	O
-	NN	O	O
a	NN	O	O
deletion	NN	O	O
spanning	NN	O	O
the	NN	O	O
third	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
-	NN	O	O
-	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
all	NN	O	O
the	NN	O	O
PKU	NN	O	B-Disease
cases	NN	O	O
among	NN	O	O
the	NN	O	O
Yemenite	NN	O	O
Jews	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
a	NN	O	O
random	NN	O	O
sample	NN	O	O
of	NN	O	O
Yemenite	NN	O	O
Jews	NN	O	O
using	NN	O	O
a	NN	O	O
molecular	NN	O	O
probe	NN	O	O
that	NN	O	O
detects	NN	O	O
the	NN	O	O
carriers	NN	O	O
of	NN	O	O
this	NN	O	O
deletion	NN	O	O
indicated	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
defective	NN	O	O
gene	NN	O	O
in	NN	O	O
this	NN	O	O
community	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
deleted	NN	O	O
PAH	NN	O	O
gene	NN	O	O
was	NN	O	O
traced	NN	O	O
to	NN	O	O
25	NN	O	O
different	NN	O	O
locations	NN	O	O
throughout	NN	O	O
Yemen	NN	O	O
,	NN	O	O
family	NN	O	O
histories	NN	O	O
and	NN	O	O
official	NN	O	O
documents	NN	O	O
of	NN	O	O
the	NN	O	O
Yemenite	NN	O	O
Jewish	NN	O	O
community	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
common	NN	O	O
ancestor	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
carriers	NN	O	O
of	NN	O	O
this	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
lived	NN	O	O
in	NN	O	O
Sana	NN	O	O
,	NN	O	O
the	NN	O	O
capital	NN	O	O
of	NN	O	O
Yemen	NN	O	O
,	NN	O	O
before	NN	O	O
the	NN	O	O
eighteenth	NN	O	O
century	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
of	NN	O	O
aspartylglucosaminuria	NN	O	B-Disease
(	NN	O	O
AGU	NN	O	B-Disease
)	NN	O	O
to	NN	O	O
marker	NN	O	O
loci	NN	O	O
on	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
4	NN	O	O
.	NN	O	O

Aspartylglucosaminuria	NN	O	B-Disease
(	NN	O	O
AGU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
deficient	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
aspartylglucosaminidase	NN	O	O
(	NN	O	O
AGA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
structural	NN	O	O
gene	NN	O	O
for	NN	O	O
AGA	NN	O	O
has	NN	O	O
been	NN	O	O
assigned	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
4q21	NN	O	O
-	NN	O	O
qter	NN	O	O
of	NN	O	O
chromosome	NN	O	O
4	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
map	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
AGU	NN	O	O
locus	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
other	NN	O	O
marker	NN	O	O
loci	NN	O	O
on	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
4	NN	O	O
using	NN	O	O
linkage	NN	O	O
analyses	NN	O	O
.	NN	O	O

Restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
alleles	NN	O	O
for	NN	O	O
the	NN	O	O
ADH2	NN	O	O
,	NN	O	O
ADH3	NN	O	O
,	NN	O	O
EGF	NN	O	O
,	NN	O	O
FG	NN	O	O
alpha	NN	O	O
and	NN	O	O
FG	NN	O	O
beta	NN	O	O
loci	NN	O	O
and	NN	O	O
blood	NN	O	O
group	NN	O	O
antigens	NN	O	O
for	NN	O	O
the	NN	O	O
MNS	NN	O	O
locus	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
12	NN	O	O
Finnish	NN	O	O
AGU	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

The	NN	O	O
heterozygous	NN	O	O
family	NN	O	O
members	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
reduced	NN	O	O
activity	NN	O	O
of	NN	O	O
AGA	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Linkage	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
using	NN	O	O
both	NN	O	O
pairwise	NN	O	O
and	NN	O	O
multipoint	NN	O	O
analyses	NN	O	O
.	NN	O	O

Loose	NN	O	O
linkage	NN	O	O
of	NN	O	O
the	NN	O	O
AGU	NN	O	B-Disease
locus	NN	O	O
to	NN	O	O
the	NN	O	O
FG	NN	O	O
and	NN	O	O
MNS	NN	O	O
loci	NN	O	O
was	NN	O	O
observed	NN	O	O
(	NN	O	O
z	NN	O	O
=	NN	O	O
1	NN	O	O
.	NN	O	O
16	NN	O	O
,	NN	O	O
z	NN	O	O
=	NN	O	O
1	NN	O	O
.	NN	O	O
39	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

Multipoint	NN	O	O
analysis	NN	O	O
to	NN	O	O
the	NN	O	O
fixed	NN	O	O
map	NN	O	O
[	NN	O	O
ADH	NN	O	O
-	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
-	NN	O	O
EGF	NN	O	O
-	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
35	NN	O	O
)	NN	O	O
-	NN	O	O
FG	NN	O	O
-	NN	O	O
(	NN	O	O
0	NN	O	O
.	NN	O	O
11	NN	O	O
)	NN	O	O
-	NN	O	O
MNS	NN	O	O
]	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
AGU	NN	O	O
locus	NN	O	O
is	NN	O	O
0	NN	O	O
.	NN	O	O

05	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

30	NN	O	O
recombination	NN	O	O
units	NN	O	O
distal	NN	O	O
to	NN	O	O
MNS	NN	O	O
(	NN	O	O
z	NN	O	O
=	NN	O	O
3	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
order	NN	O	O
cen	NN	O	O
-	NN	O	O
ADH	NN	O	O
-	NN	O	O
EGF	NN	O	O
-	NN	O	O
FG	NN	O	O
-	NN	O	O
MNS	NN	O	O
-	NN	O	O
AGU	NN	O	O
is	NN	O	O
35	NN	O	O
times	NN	O	O
more	NN	O	O
likely	NN	O	O
than	NN	O	O
the	NN	O	O
next	NN	O	O
best	NN	O	O
order	NN	O	O
cen	NN	O	O
-	NN	O	O
ADH	NN	O	O
-	NN	O	O
EGF	NN	O	O
-	NN	O	O
AGU	NN	O	O
-	NN	O	O
FG	NN	O	O
-	NN	O	O
MNS	NN	O	O
.	NN	O	O

Linkage	NN	O	O
relationships	NN	O	O
of	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
C1	NN	O	O
gene	NN	O	O
and	NN	O	O
a	NN	O	O
cytochrome	NN	O	O
P450	NN	O	O
gene	NN	O	O
(	NN	O	O
CYP2A	NN	O	O
)	NN	O	O
to	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
genetic	NN	O	O
linkage	NN	O	O
of	NN	O	O
two	NN	O	O
markers	NN	O	O
,	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
C1	NN	O	O
(	NN	O	O
APOC1	NN	O	O
)	NN	O	O
gene	NN	O	O
and	NN	O	O
a	NN	O	O
cytochrome	NN	O	O
P450	NN	O	O
(	NN	O	O
CYP2A	NN	O	O
)	NN	O	O
gene	NN	O	O
,	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

A	NN	O	O
peak	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
9	NN	O	O
.	NN	O	O

29	NN	O	O
at	NN	O	O
2	NN	O	O
cM	NN	O	O
was	NN	O	O
observed	NN	O	O
for	NN	O	O
APOC1	NN	O	O
-	NN	O	O
DM	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
8	NN	O	O
.	NN	O	O

55	NN	O	O
at	NN	O	O
4	NN	O	O
cM	NN	O	O
for	NN	O	O
CYP2A	NN	O	O
-	NN	O	O
DM	NN	O	O
.	NN	O	O

These	NN	O	O
two	NN	O	O
markers	NN	O	O
also	NN	O	O
show	NN	O	O
close	NN	O	O
linkage	NN	O	O
to	NN	O	O
each	NN	O	O
other	NN	O	O
(	NN	O	O
theta	NN	O	O
max	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
,	NN	O	O
Zmax	NN	O	O
=	NN	O	O
9	NN	O	O
.	NN	O	O
09	NN	O	O
)	NN	O	O
.	NN	O	O

From	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
genotypes	NN	O	O
of	NN	O	O
the	NN	O	O
recombinant	NN	O	O
individuals	NN	O	O
,	NN	O	O
CYP2A	NN	O	O
appears	NN	O	O
to	NN	O	O
map	NN	O	O
proximal	NN	O	O
to	NN	O	O
DM	NN	O	O
because	NN	O	O
in	NN	O	O
one	NN	O	O
recombinant	NN	O	O
individual	NN	O	O
CYP2A	NN	O	O
,	NN	O	O
APOC2	NN	O	O
and	NN	O	O
CKMM	NN	O	O
had	NN	O	O
all	NN	O	O
recombined	NN	O	O
with	NN	O	O
DM	NN	O	O
.	NN	O	O

Evidence	NN	O	O
from	NN	O	O
another	NN	O	O
CYP2A	NN	O	O
-	NN	O	O
DM	NN	O	O
recombinant	NN	O	O
individual	NN	O	O
places	NN	O	O
CYP2A	NN	O	O
proximal	NN	O	O
to	NN	O	O
APOC2	NN	O	O
and	NN	O	O
CKMM	NN	O	O
.	NN	O	O

Localisation	NN	O	O
of	NN	O	O
CYP2A	NN	O	O
on	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
also	NN	O	O
suggests	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
proximal	NN	O	O
to	NN	O	O
DM	NN	O	O
and	NN	O	O
APOC2	NN	O	O
/	NN	O	O
C1	NN	O	O
/	NN	O	O
E	NN	O	O
gene	NN	O	O
cluster	NN	O	O
.	NN	O	O

Genetic	NN	O	O
linkage	NN	O	O
map	NN	O	O
of	NN	O	O
six	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
markers	NN	O	O
around	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
on	NN	O	O
chromosome	NN	O	O
5	NN	O	O
.	NN	O	O

A	NN	O	O
genetic	NN	O	O
linkage	NN	O	O
map	NN	O	O
of	NN	O	O
six	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
markers	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
(	NN	O	O
APC	NN	O	O
)	NN	O	O
for	NN	O	O
familial	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
on	NN	O	O
chromosome	NN	O	O
5q	NN	O	O
is	NN	O	O
reported	NN	O	O
.	NN	O	O

One	NN	O	O
hundred	NN	O	O
fifty	NN	O	O
-	NN	O	O
five	NN	O	O
typed	NN	O	O
members	NN	O	O
of	NN	O	O
nine	NN	O	O
FAP	NN	O	B-Disease
kindred	NN	O	O
provided	NN	O	O
more	NN	O	O
than	NN	O	O
90	NN	O	O
meioses	NN	O	O
for	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
crucial	NN	O	O
recombination	NN	O	O
events	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
which	NN	O	O
are	NN	O	O
informative	NN	O	O
at	NN	O	O
three	NN	O	O
or	NN	O	O
more	NN	O	O
loci	NN	O	O
,	NN	O	O
allowing	NN	O	O
confident	NN	O	O
ordering	NN	O	O
of	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
map	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
,	NN	O	O
with	NN	O	O
all	NN	O	O
families	NN	O	O
showing	NN	O	O
linkage	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
chromosome	NN	O	O
5	NN	O	O
marker	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Recombination	NN	O	O
data	NN	O	O
and	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
support	NN	O	O
a	NN	O	O
locus	NN	O	O
order	NN	O	O
of	NN	O	O
centromere	NN	O	O
-	NN	O	O
pi	NN	O	O
227	NN	O	O
-	NN	O	O
C11P11	NN	O	O
-	NN	O	O
ECB27	NN	O	O
-	NN	O	O
L5	NN	O	O
.	NN	O	O

62	NN	O	O
-	NN	O	O
APC	NN	O	O
-	NN	O	O
EF5	NN	O	O
62	NN	O	O
-	NN	O	O
APC	NN	O	O
-	NN	O	O
EF5	NN	O	O
.	NN	O	O

44	NN	O	O
-	NN	O	O
YN5	NN	O	O
44	NN	O	O
-	NN	O	O
YN5	NN	O	O
.	NN	O	O

48	NN	O	O
-	NN	O	O
telomer	NN	O	O
e	NN	O	O
,	NN	O	O
although	NN	O	O
EF5	NN	O	O
.	NN	O	O

44	NN	O	O
could	NN	O	O
lie	NN	O	O
in	NN	O	O
the	NN	O	O
interval	NN	O	O
L5	NN	O	O
.	NN	O	O

62	NN	O	O
-	NN	O	O
APC	NN	O	O
or	NN	O	O
ECB27	NN	O	O
-	NN	O	O
L5	NN	O	O
.	NN	O	O

62	NN	O	O
.	NN	O	O

No	NN	O	O
recombinants	NN	O	O
were	NN	O	O
identified	NN	O	O
between	NN	O	O
APC	NN	O	O
and	NN	O	O
either	NN	O	O
EF5	NN	O	O
.	NN	O	O

44	NN	O	O
or	NN	O	O
YN5	NN	O	O
.	NN	O	O

48	NN	O	O
,	NN	O	O
but	NN	O	O
published	NN	O	O
deletion	NN	O	O
mapping	NN	O	O
in	NN	O	O
colorectal	NN	O	B-Disease
carcinomas	NN	O	I-Disease
and	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
FAP	NN	O	B-Disease
suggest	NN	O	O
that	NN	O	O
YN5	NN	O	O
.	NN	O	O

48	NN	O	O
is	NN	O	O
1	NN	O	O
-	NN	O	O
3	NN	O	O
cM	NN	O	O
from	NN	O	O
APC	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
suggests	NN	O	O
that	NN	O	O
YN5	NN	O	O
.	NN	O	O

48	NN	O	O
and	NN	O	O
L5	NN	O	O
.	NN	O	O

62	NN	O	O
delineate	NN	O	O
a	NN	O	O
small	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
within	NN	O	O
which	NN	O	O
the	NN	O	O
EF5	NN	O	O
.	NN	O	O

44	NN	O	O
locus	NN	O	O
lies	NN	O	O
very	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
not	NN	O	O
only	NN	O	O
allow	NN	O	O
use	NN	O	O
of	NN	O	O
flanking	NN	O	O
markers	NN	O	O
for	NN	O	O
presymptomatic	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
FAP	NN	O	B-Disease
but	NN	O	O
also	NN	O	O
provide	NN	O	O
a	NN	O	O
high	NN	O	O
-	NN	O	O
density	NN	O	O
map	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
for	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
itself	NN	O	O
and	NN	O	O
for	NN	O	O
further	NN	O	O
assessment	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
sporadic	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Serum	NN	O	O
amyloid	NN	O	O
A	NN	O	O
and	NN	O	O
P	NN	O	O
protein	NN	O	O
genes	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
serum	NN	O	O
amyloid	NN	O	O
A	NN	O	O
(	NN	O	O
SAA	NN	O	O
)	NN	O	O
and	NN	O	O
P	NN	O	O
(	NN	O	O
APCS	NN	O	O
)	NN	O	O
genes	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
MEF	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
SAA	NN	O	O
and	NN	O	O
APCS	NN	O	O
in	NN	O	O
MEF	NN	O	B-Disease
and	NN	O	O
MEF	NN	O	B-Disease
-	NN	O	I-Disease
amyloidosis	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
studied	NN	O	O
17	NN	O	O
informative	NN	O	O
families	NN	O	O
(	NN	O	O
15	NN	O	O
Armenians	NN	O	O
,	NN	O	O
2	NN	O	O
non	NN	O	O
-	NN	O	O
Ashkenazi	NN	O	O
Jews	NN	O	O
)	NN	O	O
and	NN	O	O
8	NN	O	O
MEF	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
amyloidosis	NN	O	B-Disease
using	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
approach	NN	O	O
.	NN	O	O

No	NN	O	O
evidence	NN	O	O
for	NN	O	O
any	NN	O	O
MEF	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
polymorphism	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
41	NN	O	O
Armenian	NN	O	O
and	NN	O	O
Jewish	NN	O	O
MEF	NN	O	B-Disease
patients	NN	O	O
tested	NN	O	O
.	NN	O	O

Our	NN	O	O
family	NN	O	O
studies	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
rule	NN	O	O
out	NN	O	O
tight	NN	O	O
linkage	NN	O	O
between	NN	O	O
SAA	NN	O	O
and	NN	O	O
MEF	NN	O	B-Disease
(	NN	O	O
lod	NN	O	O
score	NN	O	O
=	NN	O	O
-	NN	O	O
2	NN	O	O
.	NN	O	O
16	NN	O	O
,	NN	O	O
theta	NN	O	O
less	NN	O	O
than	NN	O	O
or	NN	O	O
equal	NN	O	O
to	NN	O	O
0	NN	O	O
.	NN	O	O
06	NN	O	O
)	NN	O	O
.	NN	O	O

For	NN	O	O
APCS	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
allele	NN	O	O
frequency	NN	O	O
in	NN	O	O
the	NN	O	O
MEF	NN	O	B-Disease
-	NN	O	I-Disease
amyloidosis	NN	O	I-Disease
patients	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
18	NN	O	O
unrelated	NN	O	O
MEF	NN	O	B-Disease
patients	NN	O	O
without	NN	O	O
amyloidosis	NN	O	B-Disease
and	NN	O	O
their	NN	O	O
33	NN	O	O
healthy	NN	O	O
parents	NN	O	O
.	NN	O	O

Some	NN	O	O
Mexican	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variants	NN	O	O
revisited	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
appears	NN	O	O
to	NN	O	O
be	NN	O	O
fairly	NN	O	O
common	NN	O	O
in	NN	O	O
Mexico	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
examined	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
three	NN	O	O
previously	NN	O	O
reported	NN	O	O
electrophoretically	NN	O	O
fast	NN	O	O
Mexican	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
,	NN	O	O
-	NN	O	O
G6PD	NN	O	O
Distrito	NN	O	O
Federal	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Tepic	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Castilla	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
of	NN	O	O
these	NN	O	O
variants	NN	O	O
,	NN	O	O
believed	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
biochemical	NN	O	O
characterization	NN	O	O
and	NN	O	O
population	NN	O	O
origin	NN	O	O
to	NN	O	O
be	NN	O	O
unique	NN	O	O
,	NN	O	O
have	NN	O	O
the	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
202	NN	O	O
and	NN	O	O
the	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
G	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
376	NN	O	O
,	NN	O	O
mutations	NN	O	O
that	NN	O	O
we	NN	O	O
now	NN	O	O
recognize	NN	O	O
to	NN	O	O
be	NN	O	O
characteristic	NN	O	O
of	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
Mexican	NN	O	O
males	NN	O	O
with	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
mutation	NN	O	O
.	NN	O	O

All	NN	O	O
five	NN	O	O
have	NN	O	O
the	NN	O	O
(	NN	O	O
NlaIII	NN	O	O
/	NN	O	O
FokI	NN	O	O
/	NN	O	O
PvuII	NN	O	O
/	NN	O	O
PstI	NN	O	O
)	NN	O	O
haplotype	NN	O	O
characteristic	NN	O	O
of	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
in	NN	O	O
Africa	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
PvuII	NN	O	O
+	NN	O	O
genotype	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
rare	NN	O	O
in	NN	O	O
Europe	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
all	NN	O	O
of	NN	O	O
these	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
genes	NN	O	O
had	NN	O	O
their	NN	O	O
ancient	NN	O	O
origin	NN	O	O
in	NN	O	O
Africa	NN	O	O
,	NN	O	O
although	NN	O	O
in	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
Mexican	NN	O	O
patients	NN	O	O
with	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
202A	NN	O	O
/	NN	O	O
376G	NN	O	O
the	NN	O	O
gene	NN	O	O
may	NN	O	O
have	NN	O	O
been	NN	O	O
imported	NN	O	O
more	NN	O	O
recently	NN	O	O
from	NN	O	O
Spain	NN	O	O
,	NN	O	O
where	NN	O	O
this	NN	O	O
variant	NN	O	O
,	NN	O	O
formerly	NN	O	O
known	NN	O	O
as	NN	O	O
G6PD	NN	O	O
Betica	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
prevalent	NN	O	O
.	NN	O	O
.	NN	O	O

Hereditary	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
C5	NN	O	I-Disease
in	NN	O	O
association	NN	O	O
with	NN	O	O
discoid	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
29	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
woman	NN	O	O
with	NN	O	O
discoid	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
had	NN	O	O
undetectable	NN	O	O
classic	NN	O	O
pathway	NN	O	O
complement	NN	O	O
activity	NN	O	O
.	NN	O	O

Hypocomplementemia	NN	O	B-Disease
was	NN	O	O
due	NN	O	O
to	NN	O	O
selective	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
C5	NN	O	I-Disease
.	NN	O	O

One	NN	O	O
of	NN	O	O
her	NN	O	O
children	NN	O	O
was	NN	O	O
also	NN	O	O
deficient	NN	O	O
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
documented	NN	O	O
case	NN	O	O
of	NN	O	O
an	NN	O	O
association	NN	O	O
between	NN	O	O
discoid	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
and	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Founder	NN	O	O
effect	NN	O	O
of	NN	O	O
a	NN	O	O
prevalent	NN	O	O
phenylketonuria	NN	O	B-Disease
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
Oriental	NN	O	O
population	NN	O	O
.	NN	O	O

A	NN	O	O
missense	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
[	NN	O	O
PAH	NN	O	O
;	NN	O	O
phenylalanine	NN	O	O
4	NN	O	O
-	NN	O	O
monooxygenase	NN	O	O
;	NN	O	O
L	NN	O	O
-	NN	O	O
phenylalanine	NN	O	O
,	NN	O	O
tetrahydrobiopterin	NN	O	O
oxygen	NN	O	O
oxidoreductase	NN	O	O
(	NN	O	O
4	NN	O	O
-	NN	O	O
hydroxylating	NN	O	O
)	NN	O	O
,	NN	O	O
EC	NN	O	O
1	NN	O	O
.	NN	O	O
14	NN	O	O
.	NN	O	O
16	NN	O	O
.	NN	O	O
1	NN	O	O
]	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
Chinese	NN	O	O
patient	NN	O	O
with	NN	O	O
classic	NN	O	O
phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

A	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
C	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
second	NN	O	O
base	NN	O	O
of	NN	O	O
codon	NN	O	O
413	NN	O	O
in	NN	O	O
exon	NN	O	O
12	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
Pro413	NN	O	O
for	NN	O	O
Arg413	NN	O	O
in	NN	O	O
the	NN	O	O
mutant	NN	O	O
protein	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
(	NN	O	O
R413P	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
negligible	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
when	NN	O	O
expressed	NN	O	O
in	NN	O	O
heterologous	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
and	NN	O	O
is	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
classic	NN	O	O
PKU	NN	O	B-Disease
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

Population	NN	O	O
genetic	NN	O	O
studies	NN	O	O
reveal	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
is	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
haplotype	NN	O	O
4	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
predominant	NN	O	O
haplotype	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
Oriental	NN	O	O
population	NN	O	O
.	NN	O	O

It	NN	O	O
accounts	NN	O	O
for	NN	O	O
13	NN	O	O
.	NN	O	O

8	NN	O	O
%	NN	O	O
of	NN	O	O
northern	NN	O	O
Chinese	NN	O	O
and	NN	O	O
27	NN	O	O
%	NN	O	O
of	NN	O	O
Japanese	NN	O	O
PKU	NN	O	B-Disease
alleles	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
rare	NN	O	O
in	NN	O	O
southern	NN	O	O
Chinese	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
2	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
absent	NN	O	O
in	NN	O	O
the	NN	O	O
Caucasian	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
unambiguously	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
occurred	NN	O	O
after	NN	O	O
racial	NN	O	O
divergence	NN	O	O
of	NN	O	O
Orientals	NN	O	O
and	NN	O	O
Caucasians	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
allele	NN	O	O
has	NN	O	O
spread	NN	O	O
throughout	NN	O	O
the	NN	O	O
Orient	NN	O	O
by	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
.	NN	O	O

Previous	NN	O	O
protein	NN	O	O
polymorphism	NN	O	O
studies	NN	O	O
in	NN	O	O
eastern	NN	O	O
Asia	NN	O	O
have	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
"	NN	O	O
northern	NN	O	O
Mongoloids	NN	O	O
"	NN	O	O
represented	NN	O	O
a	NN	O	O
founding	NN	O	O
population	NN	O	O
in	NN	O	O
Asia	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
in	NN	O	O
that	NN	O	O
the	NN	O	O
PKU	NN	O	B-Disease
mutation	NN	O	O
might	NN	O	O
have	NN	O	O
occurred	NN	O	O
in	NN	O	O
northern	NN	O	O
Mongoloids	NN	O	O
and	NN	O	O
subsequently	NN	O	O
spread	NN	O	O
to	NN	O	O
the	NN	O	O
Chinese	NN	O	O
and	NN	O	O
Japanese	NN	O	O
populations	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
(	NN	O	O
GGM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
manifesting	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
weeks	NN	O	O
of	NN	O	O
life	NN	O	O
and	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
selective	NN	O	O
failure	NN	O	O
to	NN	O	O
absorb	NN	O	O
dietary	NN	O	O
glucose	NN	O	O
and	NN	O	O
galactose	NN	O	O
from	NN	O	O
the	NN	O	O
intestine	NN	O	O
.	NN	O	O

The	NN	O	O
consequent	NN	O	O
severe	NN	O	O
diarrhoea	NN	O	B-Disease
and	NN	O	O
dehydration	NN	O	B-Disease
are	NN	O	O
usually	NN	O	O
fatal	NN	O	O
unless	NN	O	O
these	NN	O	O
sugars	NN	O	O
are	NN	O	O
eliminated	NN	O	O
from	NN	O	O
the	NN	O	O
diet	NN	O	O
.	NN	O	O

Intestinal	NN	O	O
biopsies	NN	O	O
of	NN	O	O
GGM	NN	O	B-Disease
patients	NN	O	O
have	NN	O	O
revealed	NN	O	O
a	NN	O	O
specific	NN	O	O
defect	NN	O	O
in	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
dependent	NN	O	O
absorption	NN	O	O
of	NN	O	O
glucose	NN	O	O
in	NN	O	O
the	NN	O	O
brush	NN	O	O
border	NN	O	O
.	NN	O	O

Normal	NN	O	O
glucose	NN	O	O
absorption	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
in	NN	O	O
the	NN	O	O
brush	NN	O	O
border	NN	O	O
membrane	NN	O	O
of	NN	O	O
the	NN	O	O
intestinal	NN	O	O
epithelium	NN	O	O
.	NN	O	O

Cellular	NN	O	O
influx	NN	O	O
is	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
transmembrane	NN	O	O
Na	NN	O	O
+	NN	O	O
electrochemical	NN	O	O
potential	NN	O	O
gradient	NN	O	O
;	NN	O	O
thereafter	NN	O	O
the	NN	O	O
sugar	NN	O	O
moves	NN	O	O
to	NN	O	O
the	NN	O	O
blood	NN	O	O
across	NN	O	O
the	NN	O	O
basolateral	NN	O	O
membrane	NN	O	O
via	NN	O	O
the	NN	O	O
facilitated	NN	O	O
glucose	NN	O	O
carrier	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
a	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
from	NN	O	O
normal	NN	O	O
human	NN	O	O
ileum	NN	O	O
and	NN	O	O
shown	NN	O	O
that	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
SGLT1	NN	O	O
,	NN	O	O
resides	NN	O	O
on	NN	O	O
the	NN	O	O
distal	NN	O	O
q	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
amplified	NN	O	O
SGLT1	NN	O	O
complementary	NN	O	O
DNA	NN	O	O
and	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
affected	NN	O	O
with	NN	O	O
GGM	NN	O	B-Disease
by	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
amplified	NN	O	O
products	NN	O	O
has	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
SGLT1	NN	O	O
which	NN	O	O
cosegregates	NN	O	O
with	NN	O	O
the	NN	O	O
GGM	NN	O	B-Disease
phenotype	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
dependent	NN	O	O
glucose	NN	O	O
transport	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
oocytes	NN	O	O
injected	NN	O	O
with	NN	O	O
this	NN	O	O
complementary	NN	O	O
RNA	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
de	NN	O	O
novo	NN	O	O
unbalanced	NN	O	O
reciprocal	NN	O	O
translocation	NN	O	O
identified	NN	O	O
as	NN	O	O
paternal	NN	O	O
in	NN	O	O
origin	NN	O	O
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Interstitial	NN	O	O
cytogenetic	NN	O	O
deletions	NN	O	O
involving	NN	O	O
the	NN	O	O
paternally	NN	O	O
derived	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
unbalanced	NN	O	O
karyotype	NN	O	O
-	NN	O	O
45	NN	O	O
,	NN	O	O
XY	NN	O	O
,	NN	O	O
-	NN	O	O
9	NN	O	O
,	NN	O	O
-	NN	O	O
15	NN	O	O
,	NN	O	O
+	NN	O	O
der	NN	O	O
(	NN	O	O
9	NN	O	O
)	NN	O	O
t	NN	O	O
(	NN	O	O
9	NN	O	O
;	NN	O	O
15	NN	O	O
)	NN	O	O
(	NN	O	O
q34	NN	O	O
;	NN	O	O
q13	NN	O	O
)	NN	O	O
.	NN	O	O

Molecular	NN	O	O
studies	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	O
probe	NN	O	O
pML34	NN	O	O
confirmed	NN	O	O
that	NN	O	O
only	NN	O	O
a	NN	O	O
single	NN	O	O
Prader	NN	O	B-Disease
Willi	NN	O	I-Disease
critical	NN	O	O
region	NN	O	O
(	NN	O	O
PWCR	NN	O	O
15q11	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
q12	NN	O	O
)	NN	O	O
copy	NN	O	O
was	NN	O	O
present	NN	O	O
.	NN	O	O

Hybridisation	NN	O	O
of	NN	O	O
patient	NN	O	O
and	NN	O	O
parental	NN	O	O
DNA	NN	O	O
with	NN	O	O
the	NN	O	O
multi	NN	O	O
-	NN	O	O
allelic	NN	O	O
probe	NN	O	O
CMW1	NN	O	O
,	NN	O	O
which	NN	O	O
maps	NN	O	O
to	NN	O	O
pter	NN	O	O
-	NN	O	O
15q13	NN	O	O
,	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
chromosome	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
translocation	NN	O	O
was	NN	O	O
paternal	NN	O	O
in	NN	O	O
origin	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
paternally	NN	O	O
-	NN	O	O
derived	NN	O	O
PWCR	NN	O	O
allele	NN	O	O
loss	NN	O	O
caused	NN	O	O
by	NN	O	O
an	NN	O	O
unbalanced	NN	O	O
translocation	NN	O	O
that	NN	O	O
has	NN	O	O
arisen	NN	O	O
de	NN	O	O
novo	NN	O	O
.	NN	O	O

Localisation	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
locus	NN	O	O
to	NN	O	O
19q13	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
19q13	NN	O	O
.	NN	O	O
3	NN	O	O
and	NN	O	O
its	NN	O	O
relationship	NN	O	O
to	NN	O	O
twelve	NN	O	O
polymorphic	NN	O	O
loci	NN	O	O
on	NN	O	O
19q	NN	O	O
.	NN	O	O

The	NN	O	O
order	NN	O	O
of	NN	O	O
fourteen	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
localised	NN	O	O
to	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
19	NN	O	O
has	NN	O	O
been	NN	O	O
established	NN	O	O
by	NN	O	O
multipoint	NN	O	O
mapping	NN	O	O
in	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
40	NN	O	O
CEPH	NN	O	O
(	NN	O	O
Centre	NN	O	O
dEtude	NN	O	O
de	NN	O	O
Polymorphisme	NN	O	O
Humain	NN	O	O
,	NN	O	O
Paris	NN	O	O
)	NN	O	O
reference	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
linkage	NN	O	O
relationship	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
locus	NN	O	O
to	NN	O	O
twelve	NN	O	O
of	NN	O	O
these	NN	O	O
markers	NN	O	O
as	NN	O	O
studied	NN	O	O
in	NN	O	O
45	NN	O	O
families	NN	O	O
with	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
genetic	NN	O	O
map	NN	O	O
is	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
localisation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
markers	NN	O	O
in	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
.	NN	O	O

Ten	NN	O	O
of	NN	O	O
the	NN	O	O
twelve	NN	O	O
markers	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
proximal	NN	O	O
to	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
two	NN	O	O
,	NN	O	O
PRKCG	NN	O	O
and	NN	O	O
D19S22	NN	O	O
,	NN	O	O
distal	NN	O	O
but	NN	O	O
at	NN	O	O
distances	NN	O	O
of	NN	O	O
approximately	NN	O	O
25	NN	O	O
cM	NN	O	O
and	NN	O	O
15	NN	O	O
cM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
closest	NN	O	O
proximal	NN	O	O
markers	NN	O	O
are	NN	O	O
APOC2	NN	O	O
(	NN	O	O
apolipoprotein	NN	O	O
C	NN	O	O
-	NN	O	O
II	NN	O	O
)	NN	O	O
and	NN	O	O
CKM	NN	O	O
(	NN	O	O
creatine	NN	O	O
kinase	NN	O	O
,	NN	O	O
muscle	NN	O	O
)	NN	O	O
approximately	NN	O	O
3	NN	O	O
cM	NN	O	O
and	NN	O	O
2	NN	O	O
cM	NN	O	O
from	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
gene	NN	O	O
respectively	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
order	NN	O	O
APOC2	NN	O	O
-	NN	O	O
CKM	NN	O	O
-	NN	O	O
DM	NN	O	O
.	NN	O	O

The	NN	O	O
distance	NN	O	O
between	NN	O	O
APOC2	NN	O	O
,	NN	O	O
CKM	NN	O	O
and	NN	O	O
DM	NN	O	B-Disease
(	NN	O	O
of	NN	O	O
the	NN	O	O
order	NN	O	O
of	NN	O	O
2	NN	O	O
million	NN	O	O
base	NN	O	O
pairs	NN	O	O
)	NN	O	O
and	NN	O	O
their	NN	O	O
known	NN	O	O
orientation	NN	O	O
should	NN	O	O
permit	NN	O	O
directional	NN	O	O
chromosome	NN	O	O
walking	NN	O	O
and	NN	O	O
jumping	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
should	NN	O	O
enable	NN	O	O
us	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
or	NN	O	O
not	NN	O	O
new	NN	O	O
markers	NN	O	O
are	NN	O	O
distal	NN	O	O
to	NN	O	O
APOC2	NN	O	O
/	NN	O	O
CKM	NN	O	O
and	NN	O	O
thus	NN	O	O
potentially	NN	O	O
flank	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O
.	NN	O	O

Localization	NN	O	O
of	NN	O	O
histidase	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
region	NN	O	O
12q22	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q24	NN	O	O
.	NN	O	O
1	NN	O	O
and	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
region	NN	O	O
10C2	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
D1	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
gene	NN	O	O
for	NN	O	O
histidase	NN	O	O
(	NN	O	O
histidine	NN	O	O
ammonia	NN	O	O
-	NN	O	O
lyase	NN	O	O
;	NN	O	O
HAL	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
enzyme	NN	O	O
deficient	NN	O	O
in	NN	O	O
histidinemia	NN	O	B-Disease
,	NN	O	O
was	NN	O	O
assigned	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	O
X	NN	O	O
mouse	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrid	NN	O	O
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
was	NN	O	O
sublocalized	NN	O	O
to	NN	O	O
region	NN	O	O
12q22	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q24	NN	O	O
.	NN	O	O

1	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
human	NN	O	O
histidase	NN	O	O
cDNA	NN	O	O
.	NN	O	O

The	NN	O	O
homologous	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
(	NN	O	O
Hal	NN	O	O
)	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
region	NN	O	O
10C2	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
D1	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
from	NN	O	O
a	NN	O	O
mouse	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
1	NN	O	O
.	NN	O	O

10	NN	O	O
Robertsonian	NN	O	O
translocation	NN	O	O
.	NN	O	O

These	NN	O	O
assignments	NN	O	O
extend	NN	O	O
the	NN	O	O
conserved	NN	O	O
syntenic	NN	O	O
region	NN	O	O
between	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O
and	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
10	NN	O	O
that	NN	O	O
includes	NN	O	O
the	NN	O	O
genes	NN	O	O
for	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
,	NN	O	O
gamma	NN	O	O
interferon	NN	O	O
,	NN	O	O
peptidase	NN	O	O
,	NN	O	O
and	NN	O	O
citrate	NN	O	O
synthase	NN	O	O
.	NN	O	O

The	NN	O	O
localization	NN	O	O
of	NN	O	O
histidase	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
10	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
histidase	NN	O	O
regulatory	NN	O	O
locus	NN	O	O
(	NN	O	O
Hsd	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
histidinemia	NN	O	O
mutation	NN	O	O
(	NN	O	O
his	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
both	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
on	NN	O	O
chromosome	NN	O	O
10	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
histidase	NN	O	O
structural	NN	O	O
gene	NN	O	O
locus	NN	O	O
.	NN	O	O

Determination	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
in	NN	O	O
17	NN	O	O
subjects	NN	O	O
.	NN	O	O

Hypoxanthine	NN	O	O
-	NN	O	O
-	NN	O	O
guanine	NN	O	O
phosphoribosyltransferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
purine	NN	O	O
salvage	NN	O	O
enzyme	NN	O	O
that	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
hypoxanthine	NN	O	O
to	NN	O	O
inosine	NN	O	O
monophosphate	NN	O	O
and	NN	O	O
guanine	NN	O	O
to	NN	O	O
guanosine	NN	O	O
monophosphate	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
of	NN	O	O
mutant	NN	O	O
HPRT	NN	O	O
proteins	NN	O	O
analyzed	NN	O	O
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
have	NN	O	O
shown	NN	O	O
a	NN	O	O
significant	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

This	NN	O	O
investigation	NN	O	O
further	NN	O	O
verifies	NN	O	O
this	NN	O	O
heterogeneity	NN	O	O
and	NN	O	O
identifies	NN	O	O
insertions	NN	O	O
,	NN	O	O
deletions	NN	O	O
,	NN	O	O
and	NN	O	O
point	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
amplified	NN	O	O
product	NN	O	O
of	NN	O	O
reverse	NN	O	O
-	NN	O	O
transcribed	NN	O	O
HPRT	NN	O	O
mRNA	NN	O	O
enabled	NN	O	O
the	NN	O	O
rapid	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
found	NN	O	O
in	NN	O	O
17	NN	O	O
previously	NN	O	O
uncharacterized	NN	O	O
cell	NN	O	O
lines	NN	O	O
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
of	NN	O	O
DNA	NN	O	O
markers	NN	O	O
at	NN	O	O
Xq28	NN	O	O
to	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
and	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
present	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
large	NN	O	O
kindred	NN	O	O
that	NN	O	O
contained	NN	O	O
patients	NN	O	O
with	NN	O	O
either	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
or	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
pedigree	NN	O	O
clearly	NN	O	O
supported	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
mode	NN	O	O
of	NN	O	O
inheritance	NN	O	O
of	NN	O	O
the	NN	O	O
nonneonatal	NN	O	O
form	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
/	NN	O	O
AMN	NN	O	B-Disease
.	NN	O	O

Analysis	NN	O	O
with	NN	O	O
DNA	NN	O	O
markers	NN	O	O
at	NN	O	O
Xq28	NN	O	O
suggested	NN	O	O
segregation	NN	O	O
of	NN	O	O
both	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
with	NN	O	O
an	NN	O	O
identical	NN	O	O
haplotype	NN	O	O
.	NN	O	O

This	NN	O	O
indicated	NN	O	O
that	NN	O	O
nonneonatal	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
are	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
at	NN	O	O
Xq28	NN	O	O
.	NN	O	O

It	NN	O	O
showed	NN	O	O
,	NN	O	O
furthermore	NN	O	O
,	NN	O	O
that	NN	O	O
phenotypic	NN	O	O
differences	NN	O	O
between	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
are	NN	O	O
not	NN	O	O
necessarily	NN	O	O
the	NN	O	O
consequence	NN	O	O
of	NN	O	O
allelic	NN	O	O
heterogeneity	NN	O	O
due	NN	O	O
to	NN	O	O
different	NN	O	O
mutations	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
maximal	NN	O	O
lod	NN	O	O
score	NN	O	O
for	NN	O	O
linkage	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
/	NN	O	O
AMN	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
the	NN	O	O
multiallelic	NN	O	O
anonymous	NN	O	O
DNA	NN	O	O
marker	NN	O	O
at	NN	O	O
DXS52	NN	O	O
was	NN	O	O
3	NN	O	O
.	NN	O	O

0	NN	O	O
at	NN	O	O
a	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

00	NN	O	O
.	NN	O	O

This	NN	O	O
made	NN	O	O
a	NN	O	O
prenatal	NN	O	O
or	NN	O	O
presymptomatic	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
heterozygote	NN	O	O
detection	NN	O	O
by	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
with	NN	O	O
this	NN	O	O
marker	NN	O	O
reliable	NN	O	O
.	NN	O	O

Skewed	NN	O	O
X	NN	O	O
inactivation	NN	O	O
in	NN	O	O
a	NN	O	O
female	NN	O	O
MZ	NN	O	O
twin	NN	O	O
results	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

One	NN	O	O
of	NN	O	O
female	NN	O	O
MZ	NN	O	O
twins	NN	O	O
presented	NN	O	O
with	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Physical	NN	O	O
examination	NN	O	O
,	NN	O	O
creatine	NN	O	O
phosphokinase	NN	O	O
levels	NN	O	O
,	NN	O	O
and	NN	O	O
muscle	NN	O	O
biopsy	NN	O	O
were	NN	O	O
consistent	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
because	NN	O	O
of	NN	O	O
her	NN	O	O
sex	NN	O	O
she	NN	O	O
was	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
limb	NN	O	B-Disease
-	NN	O	I-Disease
girdle	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

With	NN	O	O
cDNA	NN	O	O
probes	NN	O	O
to	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
deletion	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
twins	NN	O	O
and	NN	O	O
their	NN	O	O
mother	NN	O	O
.	NN	O	O

The	NN	O	O
de	NN	O	O
novo	NN	O	O
mutation	NN	O	O
which	NN	O	O
arose	NN	O	O
in	NN	O	O
the	NN	O	O
mother	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
novel	NN	O	O
junction	NN	O	O
fragments	NN	O	O
generated	NN	O	O
by	NN	O	O
HindIII	NN	O	O
,	NN	O	O
PstI	NN	O	O
,	NN	O	O
or	NN	O	O
TaqI	NN	O	O
when	NN	O	O
probed	NN	O	O
with	NN	O	O
cDNA8	NN	O	O
.	NN	O	O

Additional	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
gene	NN	O	O
deletion	NN	O	O
was	NN	O	O
given	NN	O	O
by	NN	O	O
novel	NN	O	O
SfiI	NN	O	O
junction	NN	O	O
fragments	NN	O	O
detected	NN	O	O
by	NN	O	O
probes	NN	O	O
p20	NN	O	O
,	NN	O	O
J	NN	O	O
-	NN	O	O
Bir	NN	O	O
,	NN	O	O
and	NN	O	O
J	NN	O	O
-	NN	O	O
66	NN	O	O
on	NN	O	O
pulsed	NN	O	O
-	NN	O	O
field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
PFGE	NN	O	O
)	NN	O	O
.	NN	O	O

Immunoblot	NN	O	O
analysis	NN	O	O
of	NN	O	O
muscle	NN	O	O
from	NN	O	O
the	NN	O	O
affected	NN	O	O
twin	NN	O	O
showed	NN	O	O
dystrophin	NN	O	O
of	NN	O	O
normal	NN	O	O
size	NN	O	O
but	NN	O	O
of	NN	O	O
reduced	NN	O	O
amount	NN	O	O
.	NN	O	O

Immunofluorescent	NN	O	O
visualization	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
revealed	NN	O	O
foci	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
-	NN	O	O
positive	NN	O	O
fibers	NN	O	O
adjacent	NN	O	O
to	NN	O	O
foci	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
-	NN	O	O
negative	NN	O	O
fibers	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
affected	NN	O	O
twin	NN	O	O
is	NN	O	O
a	NN	O	O
manifesting	NN	O	O
carrier	NN	O	O
of	NN	O	O
an	NN	O	O
abnormal	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
her	NN	O	O
myopathy	NN	O	B-Disease
being	NN	O	O
a	NN	O	O
direct	NN	O	O
result	NN	O	O
of	NN	O	O
underexpression	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
.	NN	O	O

Cytogenetic	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
normal	NN	O	O
karyotypes	NN	O	O
,	NN	O	O
eliminating	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
translocation	NN	O	O
affecting	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
function	NN	O	O
.	NN	O	O

Both	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
and	NN	O	O
DNA	NN	O	O
fingerprint	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
each	NN	O	O
twin	NN	O	O
has	NN	O	O
two	NN	O	O
different	NN	O	O
X	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
eliminating	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
uniparental	NN	O	B-Disease
disomy	NN	O	I-Disease
as	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
expression	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
methylation	NN	O	O
differences	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
and	NN	O	O
maternal	NN	O	O
X	NN	O	O
chromosomes	NN	O	O
in	NN	O	O
these	NN	O	O
MZ	NN	O	O
twins	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
uneven	NN	O	O
lyonization	NN	O	O
(	NN	O	O
X	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
)	NN	O	O
as	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanism	NN	O	O
for	NN	O	O
disease	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
female	NN	O	O
.	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
and	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
associated	NN	O	O
with	NN	O	O
partial	NN	O	O
adrenal	NN	O	B-Disease
insufficiency	NN	O	I-Disease
in	NN	O	O
three	NN	O	O
generations	NN	O	O
of	NN	O	O
a	NN	O	O
kindred	NN	O	O
.	NN	O	O

Four	NN	O	O
cases	NN	O	O
of	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
one	NN	O	O
case	NN	O	O
of	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
developed	NN	O	O
in	NN	O	O
a	NN	O	O
kindred	NN	O	O
over	NN	O	O
three	NN	O	O
generations	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
AMN	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
clinical	NN	O	O
variant	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

Pituitary	NN	O	O
-	NN	O	O
adrenal	NN	O	O
function	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
in	NN	O	O
10	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
including	NN	O	O
two	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
four	NN	O	O
females	NN	O	O
identified	NN	O	O
as	NN	O	O
carriers	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
/	NN	O	O
AMN	NN	O	B-Disease
.	NN	O	O

No	NN	O	O
pituitary	NN	O	B-Disease
-	NN	O	I-Disease
adrenal	NN	O	I-Disease
abnormality	NN	O	I-Disease
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
carriers	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
basal	NN	O	O
morning	NN	O	O
plasma	NN	O	O
adrenocorticotropic	NN	O	O
hormone	NN	O	O
(	NN	O	O
ACTH	NN	O	O
)	NN	O	O
levels	NN	O	O
were	NN	O	O
markedly	NN	O	O
elevated	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
males	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
,	NN	O	O
despite	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
they	NN	O	O
had	NN	O	O
no	NN	O	O
clinical	NN	O	O
signs	NN	O	O
of	NN	O	O
adrenal	NN	O	B-Disease
insufficiency	NN	O	I-Disease
and	NN	O	O
that	NN	O	O
morning	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
and	NN	O	O
their	NN	O	O
response	NN	O	O
to	NN	O	O
maximal	NN	O	O
exogenous	NN	O	O
ACTH	NN	O	O
stimulation	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
integrated	NN	O	O
24	NN	O	O
-	NN	O	O
hour	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
administration	NN	O	O
were	NN	O	O
also	NN	O	O
subnormal	NN	O	O
in	NN	O	O
these	NN	O	O
two	NN	O	O
cases	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
people	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
may	NN	O	O
have	NN	O	O
subclinical	NN	O	O
partial	NN	O	O
adrenocrotical	NN	O	B-Disease
insufficiency	NN	O	I-Disease
.	NN	O	O

No	NN	O	O
other	NN	O	O
endocrinologic	NN	O	B-Disease
dysfunction	NN	O	I-Disease
was	NN	O	O
identified	NN	O	O
.	NN	O	O
.	NN	O	O

Regional	NN	O	O
localisation	NN	O	O
of	NN	O	O
the	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
locus	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
9q13	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
assigned	NN	O	O
the	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
locus	NN	O	O
(	NN	O	O
FRDA	NN	O	O
)	NN	O	O
to	NN	O	O
chromosome	NN	O	O
9	NN	O	O
;	NN	O	O
the	NN	O	O
current	NN	O	O
maximal	NN	O	O
lod	NN	O	O
score	NN	O	O
between	NN	O	O
FRDA	NN	O	O
and	NN	O	O
MCT112	NN	O	O
(	NN	O	O
D9S15	NN	O	O
)	NN	O	O
is	NN	O	O
greater	NN	O	O
than	NN	O	O
50	NN	O	O
at	NN	O	O
a	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
of	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

The	NN	O	O
physical	NN	O	O
assignment	NN	O	O
of	NN	O	O
the	NN	O	O
locus	NN	O	O
defined	NN	O	O
by	NN	O	O
MCT112	NN	O	O
,	NN	O	O
and	NN	O	O
hence	NN	O	O
FRDA	NN	O	O
,	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
determined	NN	O	O
,	NN	O	O
although	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
MCT112	NN	O	O
with	NN	O	O
other	NN	O	O
chromosome	NN	O	O
9	NN	O	O
markers	NN	O	O
inferred	NN	O	O
a	NN	O	O
location	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
centromere	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridisation	NN	O	O
with	NN	O	O
MCT112	NN	O	O
,	NN	O	O
a	NN	O	O
corresponding	NN	O	O
cosmid	NN	O	O
MJ1	NN	O	O
,	NN	O	O
and	NN	O	O
DR47	NN	O	O
(	NN	O	O
D9S5	NN	O	O
)	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
mapping	NN	O	O
studies	NN	O	O
on	NN	O	O
hybrid	NN	O	O
cell	NN	O	O
panels	NN	O	O
,	NN	O	O
to	NN	O	O
define	NN	O	O
more	NN	O	O
precisely	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
in	NN	O	O
situ	NN	O	O
location	NN	O	O
of	NN	O	O
all	NN	O	O
three	NN	O	O
probes	NN	O	O
is	NN	O	O
9q13	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
distal	NN	O	O
to	NN	O	O
the	NN	O	O
variable	NN	O	O
heterochromatin	NN	O	O
region	NN	O	O
.	NN	O	O

Physical	NN	O	O
assignment	NN	O	O
of	NN	O	O
FRDA	NN	O	O
will	NN	O	O
allow	NN	O	O
us	NN	O	O
to	NN	O	O
identify	NN	O	O
hybrid	NN	O	O
cell	NN	O	O
lines	NN	O	O
containing	NN	O	O
the	NN	O	O
mutated	NN	O	O
gene	NN	O	O
.	NN	O	O

Increased	NN	O	O
high	NN	O	O
-	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
levels	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
common	NN	O	O
cholesteryl	NN	O	O
-	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
gene	NN	O	O
mutation	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
cholesteryl	NN	O	O
-	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
(	NN	O	O
CETP	NN	O	O
)	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
transfer	NN	O	O
of	NN	O	O
cholesteryl	NN	O	O
esters	NN	O	O
from	NN	O	O
high	NN	O	O
-	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
to	NN	O	O
other	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
described	NN	O	O
a	NN	O	O
Japanese	NN	O	O
family	NN	O	O
with	NN	O	O
increased	NN	O	O
HDL	NN	O	O
levels	NN	O	O
and	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
a	NN	O	O
splicing	NN	O	O
defect	NN	O	O
of	NN	O	O
the	NN	O	O
CETP	NN	O	O
gene	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
frequency	NN	O	O
and	NN	O	O
phenotype	NN	O	O
of	NN	O	O
this	NN	O	O
condition	NN	O	O
,	NN	O	O
we	NN	O	O
screened	NN	O	O
11	NN	O	O
additional	NN	O	O
families	NN	O	O
with	NN	O	O
high	NN	O	O
HDL	NN	O	O
levels	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
radioimmunoassay	NN	O	O
for	NN	O	O
CETP	NN	O	O
and	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
the	NN	O	O
same	NN	O	O
CETP	NN	O	O
gene	NN	O	O
mutation	NN	O	O
in	NN	O	O
four	NN	O	O
families	NN	O	O
from	NN	O	O
three	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
Japan	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
restriction	NN	O	O
-	NN	O	O
fragment	NN	O	O
-	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
CETP	NN	O	O
allele	NN	O	O
showed	NN	O	O
that	NN	O	O
all	NN	O	O
probands	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
identical	NN	O	O
haplotype	NN	O	O
.	NN	O	O

Family	NN	O	O
members	NN	O	O
homozygous	NN	O	O
for	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
n	NN	O	O
=	NN	O	O
10	NN	O	O
)	NN	O	O
had	NN	O	O
moderate	NN	O	O
hypercholesterolemia	NN	O	B-Disease
(	NN	O	O
mean	NN	O	O
total	NN	O	O
cholesterol	NN	O	O
level	NN	O	O
[	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
SD	NN	O	O
]	NN	O	O
,	NN	O	O
7	NN	O	O
.	NN	O	O
01	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
83	NN	O	O
mmol	NN	O	O
per	NN	O	O
liter	NN	O	O
)	NN	O	O
,	NN	O	O
markedly	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
HDL	NN	O	O
cholesterol	NN	O	O
(	NN	O	O
4	NN	O	O
.	NN	O	O
24	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O
01	NN	O	O
mmol	NN	O	O
per	NN	O	O
liter	NN	O	O
)	NN	O	O
and	NN	O	O
apolipoprotein	NN	O	O
A	NN	O	O
-	NN	O	O
I	NN	O	O
,	NN	O	O
and	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
low	NN	O	O
-	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
cholesterol	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
99	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
80	NN	O	O
mmol	NN	O	O
per	NN	O	O
liter	NN	O	O
)	NN	O	O
and	NN	O	O
apolipoprotein	NN	O	O
B	NN	O	O
.	NN	O	O

Members	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
deficiency	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
20	NN	O	O
)	NN	O	O
,	NN	O	O
whose	NN	O	O
CETP	NN	O	O
levels	NN	O	O
were	NN	O	O
in	NN	O	O
the	NN	O	O
lower	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
,	NN	O	O
had	NN	O	O
moderately	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
HDL	NN	O	O
cholesterol	NN	O	O
and	NN	O	O
apolipoprotein	NN	O	O
A	NN	O	O
-	NN	O	O
I	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
ratio	NN	O	O
of	NN	O	O
HDL	NN	O	O
subclass	NN	O	O
2	NN	O	O
to	NN	O	O
HDL	NN	O	O
subclass	NN	O	O
3	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
unaffected	NN	O	O
family	NN	O	O
members	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
5	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
8	NN	O	O
vs	NN	O	O
.	NN	O	O
0	NN	O	O
.	NN	O	O
7	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
4	NN	O	O
)	NN	O	O
.	NN	O	O

CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
six	NN	O	O
unrelated	NN	O	O
subjects	NN	O	O
with	NN	O	O
elevated	NN	O	O
HDL	NN	O	O
cholesterol	NN	O	O
levels	NN	O	O
who	NN	O	O
were	NN	O	O
from	NN	O	O
different	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
.	NN	O	O

CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
frequent	NN	O	O
cause	NN	O	O
of	NN	O	O
increased	NN	O	O
HDL	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
population	NN	O	O
of	NN	O	O
Japan	NN	O	O
,	NN	O	O
possibly	NN	O	O
because	NN	O	O
of	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
that	NN	O	O
we	NN	O	O
observed	NN	O	O
in	NN	O	O
heterozygotes	NN	O	O
suggest	NN	O	O
that	NN	O	O
CETP	NN	O	O
normally	NN	O	O
plays	NN	O	O
a	NN	O	O
part	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
levels	NN	O	O
of	NN	O	O
HDL	NN	O	O
subclass	NN	O	O
2	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
premature	NN	O	B-Disease
atherosclerosis	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
families	NN	O	O
with	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
the	NN	O	O
lipoprotein	NN	O	O
profile	NN	O	O
of	NN	O	O
persons	NN	O	O
with	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
potentially	NN	O	O
antiatherogenic	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
life	NN	O	O
span	NN	O	O
.	NN	O	O

Further	NN	O	O
mapping	NN	O	O
of	NN	O	O
an	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
locus	NN	O	O
to	NN	O	O
the	NN	O	O
chromosome	NN	O	O
11q23	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
mapped	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
ataxia	NN	O	B-Disease
-	NN	O	I-Disease
telangiectasia	NN	O	I-Disease
group	NN	O	O
A	NN	O	O
(	NN	O	O
ATA	NN	O	O
)	NN	O	O
to	NN	O	O
chromosome	NN	O	O
11q22	NN	O	O
-	NN	O	O
23	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
,	NN	O	O
using	NN	O	O
the	NN	O	O
genetic	NN	O	O
markers	NN	O	O
THY1	NN	O	O
and	NN	O	O
pYNB3	NN	O	O
.	NN	O	O

12	NN	O	O
(	NN	O	O
D11S144	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
likely	NN	O	O
order	NN	O	O
was	NN	O	O
cent	NN	O	O
-	NN	O	O
AT	NN	O	O
-	NN	O	O
S144	NN	O	O
-	NN	O	O
THY1	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
paper	NN	O	O
describes	NN	O	O
further	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
AT	NN	O	B-Disease
locus	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
10	NN	O	O
markers	NN	O	O
that	NN	O	O
span	NN	O	O
approximately	NN	O	O
60	NN	O	O
cM	NN	O	O
in	NN	O	O
the	NN	O	O
11q22	NN	O	O
-	NN	O	O
23	NN	O	O
region	NN	O	O
centered	NN	O	O
around	NN	O	O
S144	NN	O	O
and	NN	O	O
THY1	NN	O	O
.	NN	O	O

Location	NN	O	O
scores	NN	O	O
indicate	NN	O	O
that	NN	O	O
three	NN	O	O
contiguous	NN	O	O
subsegments	NN	O	O
within	NN	O	O
the	NN	O	O
[	NN	O	O
S144	NN	O	O
-	NN	O	O
THY1	NN	O	O
]	NN	O	O
segment	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
three	NN	O	O
contiguous	NN	O	O
segments	NN	O	O
telomeric	NN	O	O
to	NN	O	O
THY1	NN	O	O
,	NN	O	O
are	NN	O	O
each	NN	O	O
unlikely	NN	O	O
to	NN	O	O
contain	NN	O	O
the	NN	O	O
AT	NN	O	B-Disease
locus	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
more	NN	O	O
centromeric	NN	O	O
[	NN	O	O
STMY	NN	O	O
-	NN	O	O
S144	NN	O	O
]	NN	O	O
segment	NN	O	O
is	NN	O	O
most	NN	O	O
likely	NN	O	O
to	NN	O	O
contain	NN	O	O
the	NN	O	O
AT	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
recent	NN	O	O
refinements	NN	O	O
in	NN	O	O
the	NN	O	O
linkage	NN	O	O
and	NN	O	O
physical	NN	O	O
maps	NN	O	O
of	NN	O	O
11q22	NN	O	O
-	NN	O	O
23	NN	O	O
,	NN	O	O
place	NN	O	O
the	NN	O	O
AT	NN	O	B-Disease
locus	NN	O	O
at	NN	O	O
11q23	NN	O	O
.	NN	O	O

Recurrent	NN	O	O
meningitis	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
C9	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
.	NN	O	O

First	NN	O	O
case	NN	O	O
of	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Europe	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
the	NN	O	O
first	NN	O	O
cases	NN	O	O
,	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
of	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Europe	NN	O	O
that	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
Swiss	NN	O	O
family	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
two	NN	O	O
members	NN	O	O
-	NN	O	O
-	NN	O	O
one	NN	O	O
with	NN	O	O
a	NN	O	O
complete	NN	O	O
deficiency	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
with	NN	O	O
approximately	NN	O	O
half	NN	O	O
-	NN	O	O
normal	NN	O	O
C9	NN	O	O
levels	NN	O	O
-	NN	O	O
-	NN	O	O
experienced	NN	O	O
bacterial	NN	O	B-Disease
meningitis	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
index	NN	O	O
patient	NN	O	O
,	NN	O	O
a	NN	O	O
56	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
white	NN	O	O
man	NN	O	O
with	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
purulent	NN	O	B-Disease
meningitis	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
23	NN	O	O
years	NN	O	O
,	NN	O	O
presented	NN	O	O
with	NN	O	O
an	NN	O	O
acute	NN	O	B-Disease
meningococcal	NN	O	I-Disease
meningitis	NN	O	I-Disease
.	NN	O	O

No	NN	O	O
impairment	NN	O	B-Disease
of	NN	O	I-Disease
cellular	NN	O	I-Disease
immunity	NN	O	I-Disease
or	NN	O	O
immunoglobulin	NN	O	B-Disease
deficiency	NN	O	I-Disease
could	NN	O	O
be	NN	O	O
found	NN	O	O
.	NN	O	O

Complement	NN	O	O
assays	NN	O	O
showed	NN	O	O
a	NN	O	O
complete	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
C9	NN	O	I-Disease
component	NN	O	I-Disease
,	NN	O	O
while	NN	O	O
the	NN	O	O
other	NN	O	O
individual	NN	O	O
component	NN	O	O
levels	NN	O	O
were	NN	O	O
normal	NN	O	O
and	NN	O	O
the	NN	O	O
hemolytic	NN	O	O
activity	NN	O	O
(	NN	O	O
measured	NN	O	O
using	NN	O	O
the	NN	O	O
CH50	NN	O	O
assay	NN	O	O
)	NN	O	O
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
reduced	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
study	NN	O	O
revealed	NN	O	O
complete	NN	O	B-Disease
C9	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
patients	NN	O	O
healthy	NN	O	O
brother	NN	O	O
and	NN	O	O
half	NN	O	O
-	NN	O	O
normal	NN	O	O
C9	NN	O	O
concentrations	NN	O	O
in	NN	O	O
his	NN	O	O
sister	NN	O	O
,	NN	O	O
his	NN	O	O
son	NN	O	O
(	NN	O	O
who	NN	O	O
also	NN	O	O
had	NN	O	O
experienced	NN	O	O
an	NN	O	O
episode	NN	O	O
of	NN	O	O
bacterial	NN	O	B-Disease
meningitis	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
his	NN	O	O
niece	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
an	NN	O	O
inherited	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
first	NN	O	O
case	NN	O	O
of	NN	O	O
recurrent	NN	O	B-Disease
meningitis	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
white	NN	O	O
patient	NN	O	O
with	NN	O	O
complete	NN	O	B-Disease
C9	NN	O	I-Disease
deficiency	NN	O	I-Disease
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
complement	NN	O	B-Disease
defect	NN	O	I-Disease
may	NN	O	O
also	NN	O	O
be	NN	O	O
a	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
bacterial	NN	O	B-Disease
,	NN	O	I-Disease
especially	NN	O	I-Disease
neisserial	NN	O	I-Disease
,	NN	O	I-Disease
infections	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
98	NN	O	O
%	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
gene	NN	O	O
deletions	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
oligonucleotide	NN	O	O
primer	NN	O	O
sequences	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
amplify	NN	O	O
eight	NN	O	O
exons	NN	O	O
plus	NN	O	O
the	NN	O	O
muscle	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
single	NN	O	O
multiplex	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
.	NN	O	O

When	NN	O	O
used	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
an	NN	O	O
existing	NN	O	O
primer	NN	O	O
set	NN	O	O
,	NN	O	O
these	NN	O	O
two	NN	O	O
multiplex	NN	O	O
reactions	NN	O	O
detect	NN	O	O
about	NN	O	O
98	NN	O	O
%	NN	O	O
of	NN	O	O
deletions	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
or	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
,	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
primers	NN	O	O
amplify	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
exons	NN	O	O
in	NN	O	O
the	NN	O	O
deletion	NN	O	O
prone	NN	O	O
"	NN	O	O
hot	NN	O	O
spot	NN	O	O
"	NN	O	O
region	NN	O	O
around	NN	O	O
exons	NN	O	O
44	NN	O	O
to	NN	O	O
53	NN	O	O
,	NN	O	O
allowing	NN	O	O
determination	NN	O	O
of	NN	O	O
deletion	NN	O	O
endpoints	NN	O	O
and	NN	O	O
prediction	NN	O	O
of	NN	O	O
mutational	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
translational	NN	O	O
reading	NN	O	O
frame	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
use	NN	O	O
of	NN	O	O
these	NN	O	O
PCR	NN	O	O
-	NN	O	O
based	NN	O	O
assays	NN	O	O
will	NN	O	O
allow	NN	O	O
deletion	NN	O	O
detection	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
for	NN	O	O
most	NN	O	O
DMD	NN	O	B-Disease
/	NN	O	O
BMD	NN	O	B-Disease
patients	NN	O	O
in	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
time	NN	O	O
required	NN	O	O
for	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
heterogeneity	NN	O	O
at	NN	O	O
the	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
locus	NN	O	O
in	NN	O	O
southern	NN	O	O
Italy	NN	O	O
:	NN	O	O
a	NN	O	O
study	NN	O	O
on	NN	O	O
a	NN	O	O
population	NN	O	O
from	NN	O	O
the	NN	O	O
Matera	NN	O	O
district	NN	O	O
.	NN	O	O

Glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
analyzed	NN	O	O
by	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
and	NN	O	O
by	NN	O	O
quantitative	NN	O	O
assay	NN	O	O
in	NN	O	O
an	NN	O	O
unselected	NN	O	O
sample	NN	O	O
of	NN	O	O
1524	NN	O	O
schoolboys	NN	O	O
from	NN	O	O
the	NN	O	O
province	NN	O	O
of	NN	O	O
Matera	NN	O	O
(	NN	O	O
Lucania	NN	O	O
)	NN	O	O
in	NN	O	O
southern	NN	O	O
Italy	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
43	NN	O	O
subjects	NN	O	O
with	NN	O	O
a	NN	O	O
G6PD	NN	O	O
variant	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
31	NN	O	O
had	NN	O	O
severe	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
nine	NN	O	O
had	NN	O	O
mild	NN	O	O
to	NN	O	O
moderate	NN	O	O
deficiency	NN	O	O
,	NN	O	O
and	NN	O	O
three	NN	O	O
had	NN	O	O
a	NN	O	O
non	NN	O	O
-	NN	O	O
deficient	NN	O	O
electrophoretic	NN	O	O
variant	NN	O	O
.	NN	O	O

The	NN	O	O
overall	NN	O	O
rate	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
2	NN	O	O
.	NN	O	O

6	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
ranging	NN	O	O
from	NN	O	O
7	NN	O	O
.	NN	O	O

2	NN	O	O
%	NN	O	O
on	NN	O	O
the	NN	O	O
Ionian	NN	O	O
Coast	NN	O	O
to	NN	O	O
zero	NN	O	O
on	NN	O	O
the	NN	O	O
eastern	NN	O	O
side	NN	O	O
of	NN	O	O
the	NN	O	O
Lucanian	NN	O	O
Apennines	NN	O	O
,	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
inversely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
distance	NN	O	O
of	NN	O	O
each	NN	O	O
town	NN	O	O
examined	NN	O	O
from	NN	O	O
the	NN	O	O
Ionian	NN	O	O
Coast	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
geographic	NN	O	O
distribution	NN	O	O
may	NN	O	O
reflect	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
gene	NN	O	O
flow	NN	O	O
from	NN	O	O
Greek	NN	O	O
settlers	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
characterization	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
population	NN	O	O
is	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
several	NN	O	O
examples	NN	O	O
of	NN	O	O
two	NN	O	O
other	NN	O	O
known	NN	O	O
polymorphic	NN	O	O
variants	NN	O	O
(	NN	O	O
G6PD	NN	O	O
Cagliari	NN	O	O
and	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
)	NN	O	O
;	NN	O	O
three	NN	O	O
new	NN	O	O
polymorphic	NN	O	O
variants	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Metaponto	NN	O	O
(	NN	O	O
class	NN	O	O
III	NN	O	O
)	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Montalbano	NN	O	O
(	NN	O	O
class	NN	O	O
III	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Pisticci	NN	O	O
(	NN	O	O
class	NN	O	O
IV	NN	O	O
)	NN	O	O
;	NN	O	O
and	NN	O	O
two	NN	O	O
sporadic	NN	O	O
variants	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Tursi	NN	O	O
(	NN	O	O
class	NN	O	O
III	NN	O	O
)	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Ferrandina	NN	O	O
(	NN	O	O
class	NN	O	O
II	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
marked	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
within	NN	O	O
a	NN	O	O
relatively	NN	O	O
narrow	NN	O	O
geographic	NN	O	O
area	NN	O	O
and	NN	O	O
they	NN	O	O
prove	NN	O	O
the	NN	O	O
presence	NN	O	O
in	NN	O	O
the	NN	O	O
Italian	NN	O	O
peninsula	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
(	NN	O	O
GdA	NN	O	O
-	NN	O	O
)	NN	O	O
regarded	NN	O	O
as	NN	O	O
characteristically	NN	O	O
African	NN	O	O
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
murine	NN	O	I-Disease
fifth	NN	O	I-Disease
complement	NN	O	I-Disease
component	NN	O	I-Disease
(	NN	O	I-Disease
C5	NN	O	I-Disease
)	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
2	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
gene	NN	O	O
deletion	NN	O	O
in	NN	O	O
a	NN	O	O
5	NN	O	O
'	NN	O	O
-	NN	O	O
exon	NN	O	O
.	NN	O	O

To	NN	O	O
ascertain	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
that	NN	O	O
causes	NN	O	O
murine	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
genomic	NN	O	O
and	NN	O	O
cDNA	NN	O	O
libraries	NN	O	O
were	NN	O	O
constructed	NN	O	O
from	NN	O	O
mouse	NN	O	O
liver	NN	O	O
DNA	NN	O	O
and	NN	O	O
mRNA	NN	O	O
employing	NN	O	O
the	NN	O	O
congenic	NN	O	O
strains	NN	O	O
B10	NN	O	O
.	NN	O	O

D2	NN	O	O
/	NN	O	O
nSnJ	NN	O	O
and	NN	O	O
B10	NN	O	O
.	NN	O	O

D2	NN	O	O
/	NN	O	O
oSnJ	NN	O	O
that	NN	O	O
are	NN	O	O
sufficient	NN	O	O
and	NN	O	O
deficient	NN	O	B-Disease
for	NN	O	I-Disease
C5	NN	O	I-Disease
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Genomic	NN	O	O
fragments	NN	O	O
were	NN	O	O
isolated	NN	O	O
which	NN	O	O
correspond	NN	O	O
to	NN	O	O
PvuII	NN	O	O
and	NN	O	O
HindIII	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
associated	NN	O	O
with	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Sequence	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
polymorphisms	NN	O	O
resulted	NN	O	O
from	NN	O	O
single	NN	O	O
base	NN	O	O
pair	NN	O	O
substitutions	NN	O	O
and	NN	O	O
that	NN	O	O
neither	NN	O	O
substitution	NN	O	O
would	NN	O	O
probably	NN	O	O
cause	NN	O	O
or	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Sequence	NN	O	O
analyses	NN	O	O
of	NN	O	O
C5	NN	O	O
sufficient	NN	O	O
and	NN	O	O
deficient	NN	O	O
cDNAs	NN	O	O
revealed	NN	O	O
a	NN	O	O
2	NN	O	O
base	NN	O	O
-	NN	O	O
pair	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
deficient	NN	O	O
cDNAs	NN	O	O
.	NN	O	O

The	NN	O	O
"	NN	O	O
TA	NN	O	O
"	NN	O	O
deletion	NN	O	O
was	NN	O	O
located	NN	O	O
near	NN	O	O
the	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	O
.	NN	O	O

This	NN	O	O
deletion	NN	O	O
shifts	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
of	NN	O	O
the	NN	O	O
C5	NN	O	O
mRNA	NN	O	O
so	NN	O	O
that	NN	O	O
the	NN	O	O
termination	NN	O	O
codon	NN	O	O
UGA	NN	O	O
is	NN	O	O
present	NN	O	O
4	NN	O	O
base	NN	O	O
pairs	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
deletion	NN	O	O
.	NN	O	O

Genomic	NN	O	O
DNA	NN	O	O
was	NN	O	O
amplified	NN	O	O
and	NN	O	O
sequenced	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
area	NN	O	O
surrounding	NN	O	O
the	NN	O	O
2	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
.	NN	O	O

Six	NN	O	O
C5	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
strains	NN	O	O
,	NN	O	O
A	NN	O	O
/	NN	O	O
HeJ	NN	O	O
,	NN	O	O
AKR	NN	O	O
/	NN	O	O
J	NN	O	O
,	NN	O	O
DBA	NN	O	O
/	NN	O	O
2J	NN	O	O
,	NN	O	O
NZB	NN	O	O
/	NN	O	O
B1NJ	NN	O	O
,	NN	O	O
SWR	NN	O	O
/	NN	O	O
J	NN	O	O
,	NN	O	O
and	NN	O	O
B10	NN	O	O
.	NN	O	O

D2	NN	O	O
/	NN	O	O
oSnJ	NN	O	O
,	NN	O	O
and	NN	O	O
four	NN	O	O
C5	NN	O	O
-	NN	O	O
sufficient	NN	O	O
strains	NN	O	O
,	NN	O	O
Balb	NN	O	O
/	NN	O	O
CJ	NN	O	O
,	NN	O	O
C57Bl	NN	O	O
/	NN	O	O
6J	NN	O	O
,	NN	O	O
DBA	NN	O	O
/	NN	O	O
1J	NN	O	O
,	NN	O	O
and	NN	O	O
B10	NN	O	O
.	NN	O	O

D2	NN	O	O
/	NN	O	O
nSnJ	NN	O	O
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

The	NN	O	O
sequencing	NN	O	O
data	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
2	NN	O	O
base	NN	O	O
pairs	NN	O	O
were	NN	O	O
deleted	NN	O	O
from	NN	O	O
the	NN	O	O
C5	NN	O	O
gene	NN	O	O
of	NN	O	O
each	NN	O	O
deficient	NN	O	O
mouse	NN	O	O
tested	NN	O	O
but	NN	O	O
not	NN	O	O
from	NN	O	O
the	NN	O	O
C5	NN	O	O
gene	NN	O	O
of	NN	O	O
any	NN	O	O
sufficient	NN	O	O
mouse	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
1	NN	O	O
)	NN	O	O
there	NN	O	O
is	NN	O	O
an	NN	O	O
identical	NN	O	O
2	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
in	NN	O	O
an	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
C5	NN	O	O
gene	NN	O	O
in	NN	O	O
several	NN	O	O
different	NN	O	O
C5	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
mouse	NN	O	O
strains	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
mRNA	NN	O	O
transcribed	NN	O	O
from	NN	O	O
the	NN	O	O
C5D	NN	O	O
gene	NN	O	O
retains	NN	O	O
this	NN	O	O
deletion	NN	O	O
;	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
this	NN	O	O
mutation	NN	O	O
should	NN	O	O
result	NN	O	O
in	NN	O	O
C5	NN	O	B-Disease
protein	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Molecular	NN	O	O
genetics	NN	O	O
of	NN	O	O
PKU	NN	O	B-Disease
in	NN	O	O
eastern	NN	O	O
Europe	NN	O	O
:	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
associated	NN	O	O
with	NN	O	O
haplotype	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
genetic	NN	O	B-Disease
disorder	NN	O	I-Disease
secondary	NN	O	O
to	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hepatic	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

Several	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
have	NN	O	O
recently	NN	O	O
been	NN	O	O
reported	NN	O	O
,	NN	O	O
and	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
RFLP	NN	O	O
haplotypes	NN	O	O
and	NN	O	O
specific	NN	O	O
mutations	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
molecular	NN	O	B-Disease
lesion	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
exon	NN	O	O
7	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
Hungarian	NN	O	O
PKU	NN	O	B-Disease
patient	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
PCR	NN	O	O
-	NN	O	O
amplified	NN	O	O
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
causes	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
Arg243	NN	O	O
to	NN	O	O
a	NN	O	O
termination	NN	O	O
codon	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
haplotype	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
two	NN	O	O
of	NN	O	O
nine	NN	O	O
mutant	NN	O	O
haplotype	NN	O	O
4	NN	O	O
alleles	NN	O	O
among	NN	O	O
Eastern	NN	O	O
Europeans	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
present	NN	O	O
among	NN	O	O
Western	NN	O	O
Europeans	NN	O	O
and	NN	O	O
Asians	NN	O	O
.	NN	O	O

The	NN	O	O
rarity	NN	O	O
of	NN	O	O
this	NN	O	O
mutant	NN	O	O
allele	NN	O	O
and	NN	O	O
its	NN	O	O
restricted	NN	O	O
geographic	NN	O	O
distribution	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
mutational	NN	O	O
event	NN	O	O
occurred	NN	O	O
recently	NN	O	O
on	NN	O	O
a	NN	O	O
normal	NN	O	O
haplotype	NN	O	O
4	NN	O	O
background	NN	O	O
in	NN	O	O
Eastern	NN	O	O
Europe	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
red	NN	O	O
-	NN	O	O
green	NN	O	O
visual	NN	O	O
pigment	NN	O	O
gene	NN	O	O
region	NN	O	O
in	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
.	NN	O	O

Although	NN	O	O
recent	NN	O	O
data	NN	O	O
established	NN	O	O
that	NN	O	O
a	NN	O	O
specific	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
synthetase	NN	O	O
is	NN	O	O
defective	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
still	NN	O	O
unidentified	NN	O	O
.	NN	O	O

The	NN	O	O
ALD	NN	O	B-Disease
locus	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
Xq28	NN	O	O
,	NN	O	O
like	NN	O	O
the	NN	O	O
red	NN	O	O
and	NN	O	O
green	NN	O	O
color	NN	O	O
pigment	NN	O	O
genes	NN	O	O
.	NN	O	O

Abnormal	NN	O	B-Disease
color	NN	O	I-Disease
vision	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
12	NN	O	O
of	NN	O	O
27	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
milder	NN	O	O
form	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
the	NN	O	O
color	NN	O	O
vision	NN	O	O
gene	NN	O	O
cluster	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
four	NN	O	O
of	NN	O	O
eight	NN	O	O
ALD	NN	O	B-Disease
kindreds	NN	O	O
.	NN	O	O

This	NN	O	O
led	NN	O	O
us	NN	O	O
to	NN	O	O
propose	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
DNA	NN	O	O
rearrangement	NN	O	O
could	NN	O	O
underlie	NN	O	O
both	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
abnormal	NN	O	B-Disease
color	NN	O	I-Disease
vision	NN	O	I-Disease
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

Study	NN	O	O
of	NN	O	O
34	NN	O	O
French	NN	O	O
ALD	NN	O	B-Disease
patients	NN	O	O
failed	NN	O	O
to	NN	O	O
reveal	NN	O	O
a	NN	O	O
higher	NN	O	O
than	NN	O	O
expected	NN	O	O
frequency	NN	O	O
of	NN	O	O
green	NN	O	O
/	NN	O	O
red	NN	O	O
visual	NN	O	O
pigment	NN	O	O
rearrangements	NN	O	O
3	NN	O	O
to	NN	O	O
the	NN	O	O
red	NN	O	O
/	NN	O	O
green	NN	O	O
color	NN	O	O
vision	NN	O	O
gene	NN	O	O
complex	NN	O	O
.	NN	O	O

The	NN	O	O
previous	NN	O	O
report	NN	O	O
of	NN	O	O
such	NN	O	O
rearrangements	NN	O	O
was	NN	O	O
based	NN	O	O
on	NN	O	O
small	NN	O	O
numbers	NN	O	O
and	NN	O	O
lack	NN	O	O
of	NN	O	O
knowledge	NN	O	O
that	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
"	NN	O	O
abnormal	NN	O	O
"	NN	O	O
color	NN	O	O
vision	NN	O	O
arrays	NN	O	O
on	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
was	NN	O	O
twice	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
expected	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
phenotypic	NN	O	B-Disease
color	NN	O	I-Disease
vision	NN	O	I-Disease
defects	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
red	NN	O	O
/	NN	O	O
green	NN	O	O
color	NN	O	O
pigment	NN	O	O
(	NN	O	O
R	NN	O	O
/	NN	O	O
GCP	NN	O	O
)	NN	O	O
region	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
pulsed	NN	O	O
-	NN	O	O
field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
in	NN	O	O
14	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
find	NN	O	O
any	NN	O	O
fragment	NN	O	O
size	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
patients	NN	O	O
and	NN	O	O
normal	NN	O	O
individuals	NN	O	O
who	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
number	NN	O	O
of	NN	O	O
pigment	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
R	NN	O	O
/	NN	O	O
GCP	NN	O	O
region	NN	O	O
was	NN	O	O
also	NN	O	O
analyzed	NN	O	O
in	NN	O	O
29	NN	O	O
French	NN	O	O
and	NN	O	O
seven	NN	O	O
North	NN	O	O
American	NN	O	O
ALD	NN	O	B-Disease
patients	NN	O	O
by	NN	O	O
using	NN	O	O
six	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
probes	NN	O	O
,	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
cosmid	NN	O	O
walk	NN	O	O
,	NN	O	O
that	NN	O	O
flank	NN	O	O
the	NN	O	O
color	NN	O	O
vision	NN	O	O
genes	NN	O	O
.	NN	O	O

No	NN	O	O
deletions	NN	O	O
were	NN	O	O
found	NN	O	O
with	NN	O	O
probes	NN	O	O
that	NN	O	O
lie	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
green	NN	O	O
pigment	NN	O	O
genes	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
previously	NN	O	O
reported	NN	O	O
ALD	NN	O	B-Disease
individuals	NN	O	O
has	NN	O	O
a	NN	O	O
long	NN	O	O
deletion	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
red	NN	O	O
pigment	NN	O	O
gene	NN	O	O
,	NN	O	O
a	NN	O	O
deletion	NN	O	O
causing	NN	O	O
blue	NN	O	O
cone	NN	O	O
monochromacy	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
and	NN	O	O
the	NN	O	O
previous	NN	O	O
findings	NN	O	O
of	NN	O	O
a	NN	O	O
45	NN	O	O
%	NN	O	O
frequency	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
color	NN	O	O
vision	NN	O	O
defects	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
AMN	NN	O	B-Disease
may	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
/	NN	O	O
AMN	NN	O	B-Disease
gene	NN	O	O
lies	NN	O	O
5	NN	O	O
to	NN	O	O
the	NN	O	O
red	NN	O	O
pigment	NN	O	O
gene	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
frequent	NN	O	O
phenotypic	NN	O	O
color	NN	O	B-Disease
vision	NN	O	I-Disease
anomalies	NN	O	I-Disease
owe	NN	O	O
their	NN	O	O
origin	NN	O	O
to	NN	O	O
deleted	NN	O	O
DNA	NN	O	O
that	NN	O	O
includes	NN	O	O
regulatory	NN	O	O
genes	NN	O	O
for	NN	O	O
color	NN	O	O
vision	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
phenotypic	NN	O	O
color	NN	O	B-Disease
vision	NN	O	I-Disease
anomalies	NN	O	I-Disease
in	NN	O	O
AMN	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
phenocopies	NN	O	O
secondary	NN	O	O
to	NN	O	O
retinal	NN	O	O
or	NN	O	O
neural	NN	O	O
involvement	NN	O	O
by	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
single	NN	O	O
case	NN	O	O
of	NN	O	O
blue	NN	O	O
cone	NN	O	O
monochromacy	NN	O	O
may	NN	O	O
therefore	NN	O	O
be	NN	O	O
a	NN	O	O
fortuitous	NN	O	O
coincidence	NN	O	O
of	NN	O	O
two	NN	O	O
diseases	NN	O	O
.	NN	O	O
.	NN	O	O

Paroxysmal	NN	O	B-Disease
nocturnal	NN	O	I-Disease
haemoglobinuria	NN	O	I-Disease
with	NN	O	O
coexisting	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
ninth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
:	NN	O	O
lack	NN	O	O
of	NN	O	O
massive	NN	O	O
haemolytic	NN	O	B-Disease
attack	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
47	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
woman	NN	O	O
with	NN	O	O
paroxysmal	NN	O	B-Disease
nocturnal	NN	O	I-Disease
haemoglobinuria	NN	O	I-Disease
(	NN	O	O
PNH	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
ninth	NN	O	I-Disease
complement	NN	O	I-Disease
component	NN	O	I-Disease
(	NN	O	O
C9	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
complement	NN	O	O
-	NN	O	O
sensitivity	NN	O	O
lysis	NN	O	O
tests	NN	O	O
,	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
her	NN	O	O
erythrocytes	NN	O	O
were	NN	O	O
markedly	NN	O	O
complement	NN	O	O
-	NN	O	O
sensitive	NN	O	O
(	NN	O	O
PNH	NN	O	O
-	NN	O	O
III	NN	O	O
)	NN	O	O
.	NN	O	O

Laser	NN	O	O
cytofluorimetry	NN	O	O
with	NN	O	O
a	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
against	NN	O	O
decay	NN	O	O
-	NN	O	O
accelerating	NN	O	O
factor	NN	O	O
(	NN	O	O
DAF	NN	O	O
)	NN	O	O
revealed	NN	O	O
that	NN	O	O
95	NN	O	O
%	NN	O	O
of	NN	O	O
her	NN	O	O
erythrocytes	NN	O	O
were	NN	O	O
DAF	NN	O	O
-	NN	O	O
negative	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
she	NN	O	O
has	NN	O	O
suffered	NN	O	O
only	NN	O	O
mild	NN	O	O
haemolysis	NN	O	B-Disease
and	NN	O	O
has	NN	O	O
never	NN	O	O
experienced	NN	O	O
massive	NN	O	O
spontaneous	NN	O	O
haemolysis	NN	O	B-Disease
.	NN	O	O

Gross	NN	O	O
haemoglobinuria	NN	O	B-Disease
and	NN	O	O
jaundice	NN	O	B-Disease
occurred	NN	O	O
only	NN	O	O
after	NN	O	O
receiving	NN	O	O
postoperative	NN	O	O
transfusion	NN	O	O
of	NN	O	O
whole	NN	O	O
blood	NN	O	O
.	NN	O	O

In	NN	O	O
her	NN	O	O
serum	NN	O	O
,	NN	O	O
C9	NN	O	O
was	NN	O	O
not	NN	O	O
detectable	NN	O	O
either	NN	O	O
by	NN	O	O
immunological	NN	O	O
or	NN	O	O
by	NN	O	O
functional	NN	O	O
assays	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
Ham	NN	O	O
test	NN	O	O
and	NN	O	O
the	NN	O	O
sugar	NN	O	O
water	NN	O	O
test	NN	O	O
using	NN	O	O
normal	NN	O	O
human	NN	O	O
serum	NN	O	O
or	NN	O	O
plasma	NN	O	O
yielded	NN	O	O
marked	NN	O	O
haemolysis	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
erythrocytes	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
patients	NN	O	O
serum	NN	O	O
or	NN	O	O
plasma	NN	O	O
was	NN	O	O
used	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
trace	NN	O	O
of	NN	O	O
lysis	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
purified	NN	O	O
human	NN	O	O
C9	NN	O	O
to	NN	O	O
her	NN	O	O
plasma	NN	O	O
fully	NN	O	O
restored	NN	O	O
haemolysis	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicated	NN	O	O
that	NN	O	O
C9	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
haemolytic	NN	O	B-Disease
attacks	NN	O	I-Disease
in	NN	O	O
patients	NN	O	O
with	NN	O	O
PNH	NN	O	B-Disease
and	NN	O	O
that	NN	O	O
characteristic	NN	O	O
haemolysis	NN	O	B-Disease
in	NN	O	O
PNH	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
tempered	NN	O	O
by	NN	O	O
coexisting	NN	O	O
C9	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Duplicational	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
locus	NN	O	O
:	NN	O	O
its	NN	O	O
frequency	NN	O	O
,	NN	O	O
distribution	NN	O	O
,	NN	O	O
origin	NN	O	O
,	NN	O	O
and	NN	O	O
phenotypegenotype	NN	O	O
correlation	NN	O	O
.	NN	O	O

Partial	NN	O	O
gene	NN	O	O
deletion	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
mutation	NN	O	O
leading	NN	O	O
to	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Partial	NN	O	O
gene	NN	O	O
duplication	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
recognized	NN	O	O
in	NN	O	O
a	NN	O	O
few	NN	O	O
cases	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
conducted	NN	O	O
a	NN	O	O
survey	NN	O	O
for	NN	O	O
duplication	NN	O	O
in	NN	O	O
72	NN	O	O
unrelated	NN	O	O
nondeletion	NN	O	O
patients	NN	O	O
,	NN	O	O
analyzed	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
with	NN	O	O
clones	NN	O	O
representing	NN	O	O
the	NN	O	O
entire	NN	O	O
DMD	NN	O	B-Disease
cDNA	NN	O	O
.	NN	O	O

With	NN	O	O
careful	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
of	NN	O	O
hybridization	NN	O	O
band	NN	O	O
intensity	NN	O	O
,	NN	O	O
10	NN	O	O
cases	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
carry	NN	O	O
a	NN	O	O
duplication	NN	O	O
of	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
a	NN	O	O
frequency	NN	O	O
of	NN	O	O
14	NN	O	O
%	NN	O	O
for	NN	O	O
nondeletion	NN	O	O
cases	NN	O	O
(	NN	O	O
10	NN	O	O
/	NN	O	O
72	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
6	NN	O	O
%	NN	O	O
for	NN	O	O
all	NN	O	O
cases	NN	O	O
(	NN	O	O
10	NN	O	O
/	NN	O	O
181	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
extent	NN	O	O
of	NN	O	O
these	NN	O	O
duplications	NN	O	O
has	NN	O	O
been	NN	O	O
characterized	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
published	NN	O	O
exon	NN	O	O
-	NN	O	O
containing	NN	O	O
HindIII	NN	O	O
fragment	NN	O	O
map	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
six	NN	O	O
of	NN	O	O
the	NN	O	O
10	NN	O	O
duplications	NN	O	O
a	NN	O	O
novel	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
that	NN	O	O
spanned	NN	O	O
the	NN	O	O
duplication	NN	O	O
junction	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
translational	NN	O	O
reading	NN	O	O
frame	NN	O	O
of	NN	O	O
mRNA	NN	O	O
has	NN	O	O
been	NN	O	O
predicted	NN	O	O
for	NN	O	O
nine	NN	O	O
duplications	NN	O	O
.	NN	O	O

A	NN	O	O
shift	NN	O	O
of	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
was	NN	O	O
predicted	NN	O	O
in	NN	O	O
four	NN	O	O
of	NN	O	O
the	NN	O	O
six	NN	O	O
DMD	NN	O	B-Disease
cases	NN	O	O
and	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
intermediate	NN	O	O
cases	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
remained	NN	O	O
uninterrupted	NN	O	O
in	NN	O	O
both	NN	O	O
BMD	NN	O	B-Disease
cases	NN	O	O
.	NN	O	O

RFLP	NN	O	O
and	NN	O	O
quantitative	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
revealed	NN	O	O
a	NN	O	O
grandpaternal	NN	O	O
origin	NN	O	O
of	NN	O	O
duplication	NN	O	O
in	NN	O	O
four	NN	O	O
families	NN	O	O
and	NN	O	O
grandmaternal	NN	O	O
origin	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
five	NN	O	O
families	NN	O	O
,	NN	O	O
the	NN	O	O
duplication	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
originate	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Unequal	NN	O	O
sister	NN	O	O
-	NN	O	O
chromatid	NN	O	O
exchange	NN	O	O
is	NN	O	O
proposed	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
these	NN	O	O
duplications	NN	O	O
.	NN	O	O
.	NN	O	O

Glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variants	NN	O	O
:	NN	O	O
Gd	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
Alexandra	NN	O	O
associated	NN	O	O
with	NN	O	O
neonatal	NN	O	B-Disease
jaundice	NN	O	I-Disease
and	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Camperdown	NN	O	O
in	NN	O	O
a	NN	O	O
young	NN	O	O
man	NN	O	O
with	NN	O	O
lamellar	NN	O	B-Disease
cataracts	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
male	NN	O	O
subjects	NN	O	O
are	NN	O	O
described	NN	O	O
,	NN	O	O
with	NN	O	O
unusual	NN	O	O
clinical	NN	O	O
presentations	NN	O	O
and	NN	O	O
with	NN	O	O
hitherto	NN	O	O
undescribed	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
,	NN	O	O
of	NN	O	O
Italian	NN	O	O
extraction	NN	O	O
,	NN	O	O
suffered	NN	O	O
from	NN	O	O
severe	NN	O	O
neonatal	NN	O	B-Disease
jaundice	NN	O	I-Disease
following	NN	O	O
maternal	NN	O	O
ingestion	NN	O	O
of	NN	O	O
fresh	NN	O	O
broad	NN	O	O
beans	NN	O	O
(	NN	O	O
Vicia	NN	O	O
fava	NN	O	O
)	NN	O	O
both	NN	O	O
prenatally	NN	O	O
and	NN	O	O
postnatally	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
enzymatic	NN	O	O
defect	NN	O	O
was	NN	O	O
much	NN	O	O
more	NN	O	O
severe	NN	O	O
in	NN	O	O
the	NN	O	O
neonatal	NN	O	O
period	NN	O	O
than	NN	O	O
on	NN	O	O
retesting	NN	O	O
in	NN	O	O
adolescence	NN	O	O
,	NN	O	O
when	NN	O	O
biochemical	NN	O	O
characterization	NN	O	O
showed	NN	O	O
unique	NN	O	O
features	NN	O	O
which	NN	O	O
justify	NN	O	O
designation	NN	O	O
as	NN	O	O
a	NN	O	O
new	NN	O	O
variant	NN	O	O
Gd	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
Alexandra	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
patient	NN	O	O
,	NN	O	O
a	NN	O	O
boy	NN	O	O
of	NN	O	O
Maltese	NN	O	O
extraction	NN	O	O
who	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
bilateral	NN	O	B-Disease
lamellar	NN	O	I-Disease
cataracts	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
4	NN	O	O
years	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
G6PD	NN	O	B-Disease
deficient	NN	O	I-Disease
only	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
survey	NN	O	O
of	NN	O	O
children	NN	O	O
of	NN	O	O
Mediterranean	NN	O	O
origin	NN	O	O
with	NN	O	O
unexplained	NN	O	O
cataract	NN	O	B-Disease
formation	NN	O	O
;	NN	O	O
he	NN	O	O
has	NN	O	O
approximately	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
,	NN	O	O
with	NN	O	O
another	NN	O	O
unique	NN	O	O
combination	NN	O	O
of	NN	O	O
biochemical	NN	O	O
characteristics	NN	O	O
which	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
its	NN	O	O
designation	NN	O	O
as	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Camperdown	NN	O	O
.	NN	O	O

Although	NN	O	O
this	NN	O	O
association	NN	O	O
may	NN	O	O
be	NN	O	O
coincidental	NN	O	O
,	NN	O	O
it	NN	O	O
prompts	NN	O	O
further	NN	O	O
attention	NN	O	O
to	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
under	NN	O	O
certain	NN	O	O
circumstances	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
may	NN	O	O
favor	NN	O	O
cataract	NN	O	B-Disease
formation	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
cases	NN	O	O
illustrate	NN	O	O
the	NN	O	O
value	NN	O	O
of	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
enzyme	NN	O	O
whenever	NN	O	O
unexpected	NN	O	O
clinical	NN	O	O
or	NN	O	O
laboratory	NN	O	O
results	NN	O	O
are	NN	O	O
obtained	NN	O	O
.	NN	O	O
.	NN	O	O

Statistical	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
stage	NN	O	O
mutation	NN	O	O
model	NN	O	O
in	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
in	NN	O	O
sporadic	NN	O	B-Disease
cerebellar	NN	O	I-Disease
haemangioblastoma	NN	O	I-Disease
and	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
age	NN	O	O
incidence	NN	O	O
curves	NN	O	O
for	NN	O	O
unilateral	NN	O	B-Disease
and	NN	O	I-Disease
bilateral	NN	O	I-Disease
retinoblastoma	NN	O	I-Disease
led	NN	O	O
Knudson	NN	O	O
to	NN	O	O
propose	NN	O	O
that	NN	O	O
hereditary	NN	O	B-Disease
tumours	NN	O	I-Disease
may	NN	O	O
arise	NN	O	O
by	NN	O	O
a	NN	O	O
single	NN	O	O
event	NN	O	O
and	NN	O	O
sporadic	NN	O	B-Disease
tumours	NN	O	I-Disease
by	NN	O	O
a	NN	O	O
two	NN	O	O
stage	NN	O	O
mutation	NN	O	O
process	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
recently	NN	O	O
that	NN	O	O
sporadic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
may	NN	O	O
arise	NN	O	O
from	NN	O	O
a	NN	O	O
two	NN	O	O
stage	NN	O	O
mutation	NN	O	O
process	NN	O	O
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
the	NN	O	O
age	NN	O	O
incidence	NN	O	O
curves	NN	O	O
for	NN	O	O
symptomatic	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
(	NN	O	O
n	NN	O	O
=	NN	O	O
26	NN	O	O
)	NN	O	O
and	NN	O	O
cerebellar	NN	O	B-Disease
haemangioblastoma	NN	O	I-Disease
(	NN	O	O
n	NN	O	O
=	NN	O	O
68	NN	O	O
)	NN	O	O
in	NN	O	O
109	NN	O	O
patients	NN	O	O
with	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	I-Disease
VHL	NN	O	I-Disease
)	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
compared	NN	O	O
them	NN	O	O
to	NN	O	O
104	NN	O	O
patients	NN	O	O
with	NN	O	O
sporadic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
and	NN	O	O
43	NN	O	O
patients	NN	O	O
with	NN	O	O
sporadic	NN	O	B-Disease
cerebellar	NN	O	I-Disease
haemangioblastoma	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
age	NN	O	O
incidence	NN	O	O
curves	NN	O	O
for	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
and	NN	O	O
cerebellar	NN	O	B-Disease
haemangioblastoma	NN	O	I-Disease
in	NN	O	O
VHL	NN	O	B-Disease
disease	NN	O	I-Disease
were	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
single	NN	O	O
mutation	NN	O	O
model	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
age	NN	O	O
incidence	NN	O	O
curves	NN	O	O
for	NN	O	O
sporadic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
and	NN	O	O
cerebellar	NN	O	B-Disease
haemangioblastoma	NN	O	I-Disease
suggested	NN	O	O
a	NN	O	O
two	NN	O	O
stage	NN	O	O
mutation	NN	O	O
process	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
functioning	NN	O	O
as	NN	O	O
a	NN	O	O
recessive	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
.	NN	O	O

Sporadic	NN	O	B-Disease
cerebellar	NN	O	I-Disease
haemangioblastoma	NN	O	I-Disease
and	NN	O	O
some	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
may	NN	O	O
arise	NN	O	O
from	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
inactivating	NN	O	O
both	NN	O	O
alleles	NN	O	O
at	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O
.	NN	O	O

Screening	NN	O	O
for	NN	O	O
carriers	NN	O	O
of	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
among	NN	O	O
Ashkenazi	NN	O	O
Jews	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
DNA	NN	O	O
-	NN	O	O
based	NN	O	O
and	NN	O	O
enzyme	NN	O	O
-	NN	O	O
based	NN	O	O
tests	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
.	NN	O	O

The	NN	O	O
prevention	NN	O	O
of	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
GM2	NN	O	B-Disease
gangliosidosis	NN	O	I-Disease
,	NN	O	I-Disease
type	NN	O	I-Disease
1	NN	O	I-Disease
)	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
this	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
compared	NN	O	O
the	NN	O	O
enzyme	NN	O	O
-	NN	O	O
based	NN	O	O
test	NN	O	O
widely	NN	O	O
used	NN	O	O
in	NN	O	O
screening	NN	O	O
for	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
with	NN	O	O
a	NN	O	O
test	NN	O	O
based	NN	O	O
on	NN	O	O
analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

We	NN	O	O
developed	NN	O	O
methods	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
three	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
that	NN	O	O
occur	NN	O	O
with	NN	O	O
high	NN	O	O
frequency	NN	O	O
among	NN	O	O
Ashkenazi	NN	O	O
Jews	NN	O	O
two	NN	O	O
mutations	NN	O	O
cause	NN	O	O
infantile	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
the	NN	O	O
third	NN	O	O
causes	NN	O	O
the	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

DNA	NN	O	O
segments	NN	O	O
containing	NN	O	O
these	NN	O	O
mutation	NN	O	O
sites	NN	O	O
were	NN	O	O
amplified	NN	O	O
with	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
analyzed	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Among	NN	O	O
62	NN	O	O
Ashkenazi	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
of	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
three	NN	O	O
specific	NN	O	O
mutations	NN	O	O
accounted	NN	O	O
for	NN	O	O
all	NN	O	O
but	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
(	NN	O	O
98	NN	O	O
percent	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
216	NN	O	O
Ashkenazi	NN	O	O
carriers	NN	O	O
identified	NN	O	O
by	NN	O	O
the	NN	O	O
enzyme	NN	O	O
test	NN	O	O
,	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
177	NN	O	O
(	NN	O	O
82	NN	O	O
percent	NN	O	O
)	NN	O	O
had	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
identified	NN	O	O
mutations	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
177	NN	O	O
,	NN	O	O
79	NN	O	O
percent	NN	O	O
had	NN	O	O
the	NN	O	O
exon	NN	O	O
11	NN	O	O
insertion	NN	O	O
mutation	NN	O	O
,	NN	O	O
18	NN	O	O
percent	NN	O	O
had	NN	O	O
the	NN	O	O
intron	NN	O	O
12	NN	O	O
splice	NN	O	O
-	NN	O	O
junction	NN	O	O
mutation	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
percent	NN	O	O
had	NN	O	O
the	NN	O	O
less	NN	O	O
severe	NN	O	O
exon	NN	O	O
7	NN	O	O
mutation	NN	O	O
associated	NN	O	O
with	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
tests	NN	O	O
in	NN	O	O
the	NN	O	O
39	NN	O	O
subjects	NN	O	O
(	NN	O	O
18	NN	O	O
percent	NN	O	O
)	NN	O	O
who	NN	O	O
were	NN	O	O
defined	NN	O	O
as	NN	O	O
carriers	NN	O	O
but	NN	O	O
in	NN	O	O
whom	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
did	NN	O	O
not	NN	O	O
identify	NN	O	O
a	NN	O	O
mutant	NN	O	O
allele	NN	O	O
were	NN	O	O
probably	NN	O	O
false	NN	O	O
positive	NN	O	O
(	NN	O	O
although	NN	O	O
there	NN	O	O
remains	NN	O	O
some	NN	O	O
possibility	NN	O	O
of	NN	O	O
unidentified	NN	O	O
mutations	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
of	NN	O	O
152	NN	O	O
persons	NN	O	O
defined	NN	O	O
as	NN	O	O
noncarriers	NN	O	O
by	NN	O	O
the	NN	O	O
enzyme	NN	O	O
-	NN	O	O
based	NN	O	O
test	NN	O	O
,	NN	O	O
1	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
carrier	NN	O	O
by	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
(	NN	O	O
i	NN	O	O
.	NN	O	O
e	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
a	NN	O	O
false	NN	O	O
negative	NN	O	O
enzyme	NN	O	O
-	NN	O	O
test	NN	O	O
result	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
.	NN	O	O

The	NN	O	O
increased	NN	O	O
specificity	NN	O	O
and	NN	O	O
predictive	NN	O	O
value	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
-	NN	O	O
based	NN	O	O
test	NN	O	O
make	NN	O	O
it	NN	O	O
a	NN	O	O
useful	NN	O	O
adjunct	NN	O	O
to	NN	O	O
the	NN	O	O
diagnostic	NN	O	O
tests	NN	O	O
currently	NN	O	O
used	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
carriers	NN	O	O
of	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Total	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
plasma	NN	O	I-Disease
cholesteryl	NN	O	I-Disease
ester	NN	O	I-Disease
transfer	NN	O	I-Disease
protein	NN	O	I-Disease
in	NN	O	O
subjects	NN	O	O
homozygous	NN	O	O
and	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
intron	NN	O	O
14	NN	O	O
splicing	NN	O	O
defect	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
cholesteryl	NN	O	B-Disease
ester	NN	O	I-Disease
transfer	NN	O	I-Disease
protein	NN	O	I-Disease
(	NN	O	I-Disease
CETP	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
4	NN	O	O
unrelated	NN	O	O
CETP	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
probands	NN	O	O
exceeded	NN	O	O
150	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
of	NN	O	O
the	NN	O	O
probands	NN	O	O
was	NN	O	O
totally	NN	O	O
deficient	NN	O	O
in	NN	O	O
CETP	NN	O	O
activity	NN	O	O
and	NN	O	O
mass	NN	O	O
.	NN	O	O

The	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
amplified	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
,	NN	O	O
using	NN	O	O
two	NN	O	O
oligonucleotide	NN	O	O
primers	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
intron	NN	O	O
12	NN	O	O
and	NN	O	O
14	NN	O	O
of	NN	O	O
the	NN	O	O
CETP	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
amplified	NN	O	O
products	NN	O	O
were	NN	O	O
directly	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Two	NN	O	O
patients	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
change	NN	O	O
at	NN	O	O
the	NN	O	O
5	NN	O	O
-	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	O
14	NN	O	O
.	NN	O	O

The	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
change	NN	O	O
would	NN	O	O
cause	NN	O	O
impaired	NN	O	O
splicing	NN	O	O
of	NN	O	O
pre	NN	O	O
-	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
two	NN	O	O
probands	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
,	NN	O	O
but	NN	O	O
totally	NN	O	O
lacked	NN	O	O
CETP	NN	O	O
.	NN	O	O

Their	NN	O	O
lipoprotein	NN	O	O
patterns	NN	O	O
were	NN	O	O
also	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
homozygotes	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
other	NN	O	O
genetic	NN	O	B-Disease
defects	NN	O	I-Disease
or	NN	O	O
metabolic	NN	O	O
factors	NN	O	O
influencing	NN	O	O
CETP	NN	O	O
expression	NN	O	O
are	NN	O	O
implicated	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
mutation	NN	O	O
may	NN	O	O
be	NN	O	O
common	NN	O	O
in	NN	O	O
human	NN	O	O
plasma	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
there	NN	O	O
could	NN	O	O
be	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
who	NN	O	O
totally	NN	O	O
lack	NN	O	O
plasma	NN	O	O
CETP	NN	O	O
and	NN	O	O
have	NN	O	O
lipoprotein	NN	O	O
profiles	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
homozygotes	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
genetics	NN	O	O
of	NN	O	O
the	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
Mediterranean	NN	O	O
variant	NN	O	O
and	NN	O	O
description	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
G6PD	NN	O	O
mutant	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Andalus1361A	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
;	NN	O	I-Disease
E	NN	O	I-Disease
.	NN	O	I-Disease
C	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
49	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
human	NN	O	O
enzymopathy	NN	O	B-Disease
;	NN	O	O
more	NN	O	O
than	NN	O	O
300	NN	O	O
different	NN	O	O
biochemical	NN	O	O
variants	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

In	NN	O	O
many	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
world	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
type	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
prevalent	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
has	NN	O	O
come	NN	O	O
to	NN	O	O
be	NN	O	O
regarded	NN	O	O
as	NN	O	O
a	NN	O	O
generic	NN	O	O
term	NN	O	O
applied	NN	O	O
to	NN	O	O
similar	NN	O	O
G6PD	NN	O	O
mutations	NN	O	O
thought	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
to	NN	O	O
represent	NN	O	O
a	NN	O	O
somewhat	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
.	NN	O	O

A	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
mutation	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
563	NN	O	O
of	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
by	NN	O	O
Vulliamy	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
same	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
by	NN	O	O
De	NN	O	O
Vita	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
in	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Sassari	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Cagliari	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
subjects	NN	O	O
had	NN	O	O
an	NN	O	O
additional	NN	O	O
mutation	NN	O	O
,	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1311	NN	O	O
,	NN	O	O
that	NN	O	O
did	NN	O	O
not	NN	O	O
produce	NN	O	O
a	NN	O	O
coding	NN	O	O
change	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
of	NN	O	O
five	NN	O	O
patients	NN	O	O
-	NN	O	O
-	NN	O	O
four	NN	O	O
of	NN	O	O
Spanish	NN	O	O
origin	NN	O	O
and	NN	O	O
one	NN	O	O
of	NN	O	O
Jewish	NN	O	O
origin	NN	O	O
-	NN	O	O
-	NN	O	O
having	NN	O	O
enzymatically	NN	O	O
documented	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
.	NN	O	O

All	NN	O	O
had	NN	O	O
both	NN	O	O
the	NN	O	O
mutation	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
563	NN	O	O
and	NN	O	O
that	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1311	NN	O	O
.	NN	O	O

A	NN	O	O
sixth	NN	O	O
sample	NN	O	O
,	NN	O	O
resembling	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
kinetically	NN	O	O
but	NN	O	O
with	NN	O	O
a	NN	O	O
slightly	NN	O	O
rapid	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
,	NN	O	O
was	NN	O	O
designated	NN	O	O
G6PD	NN	O	O
Andalus	NN	O	O
and	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
different	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1361	NN	O	O
,	NN	O	O
producing	NN	O	O
an	NN	O	O
arginine	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
histidine	NN	O	O
substitution	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
is	NN	O	O
,	NN	O	O
after	NN	O	O
all	NN	O	O
,	NN	O	O
relatively	NN	O	O
homogeneous	NN	O	O
.	NN	O	O

A	NN	O	O
normal	NN	O	O
male	NN	O	O
with	NN	O	O
an	NN	O	O
inherited	NN	O	O
deletion	NN	O	O
of	NN	O	O
one	NN	O	O
exon	NN	O	O
within	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
two	NN	O	O
brothers	NN	O	O
with	NN	O	O
identical	NN	O	O
inherited	NN	O	O
deletions	NN	O	O
of	NN	O	O
one	NN	O	O
single	NN	O	O
exon	NN	O	O
within	NN	O	O
the	NN	O	O
middle	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
;	NN	O	O
one	NN	O	O
brother	NN	O	O
has	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
diagnosed	NN	O	O
at	NN	O	O
11	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
older	NN	O	O
brother	NN	O	O
is	NN	O	O
normal	NN	O	O
at	NN	O	O
18	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
genetic	NN	O	O
counselling	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
diseased	NN	O	O
human	NN	O	O
muscle	NN	O	O
.	NN	O	O

A	NN	O	O
probe	NN	O	O
for	NN	O	O
the	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
study	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	O
muscle	NN	O	O
,	NN	O	O
myogenic	NN	O	O
cell	NN	O	O
cultures	NN	O	O
,	NN	O	O
and	NN	O	O
muscle	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

Expression	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
RNA	NN	O	O
from	NN	O	O
normal	NN	O	O
fetal	NN	O	O
muscle	NN	O	O
,	NN	O	O
adult	NN	O	O
cardiac	NN	O	O
and	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
,	NN	O	O
and	NN	O	O
cultured	NN	O	O
muscle	NN	O	O
after	NN	O	O
myoblast	NN	O	O
fusion	NN	O	O
.	NN	O	O

In	NN	O	O
DMD	NN	O	B-Disease
muscle	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
was	NN	O	O
also	NN	O	O
revealed	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
RNA	NN	O	O
hybridization	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
regenerating	NN	O	O
muscle	NN	O	O
fibers	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
and	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
deletions	NN	O	O
within	NN	O	O
the	NN	O	O
deletion	NN	O	O
-	NN	O	O
rich	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Eighty	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
or	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
major	NN	O	O
deletion	NN	O	O
-	NN	O	O
rich	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

This	NN	O	O
region	NN	O	O
includes	NN	O	O
the	NN	O	O
last	NN	O	O
five	NN	O	O
exons	NN	O	O
detected	NN	O	O
by	NN	O	O
cDNA5b	NN	O	O
-	NN	O	O
7	NN	O	O
,	NN	O	O
all	NN	O	O
exons	NN	O	O
detected	NN	O	O
by	NN	O	O
cDNA8	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
first	NN	O	O
two	NN	O	O
exons	NN	O	O
detected	NN	O	O
by	NN	O	O
cDNA9	NN	O	O
.	NN	O	O

These	NN	O	O
80	NN	O	O
individuals	NN	O	O
account	NN	O	O
for	NN	O	O
approximately	NN	O	O
75	NN	O	O
%	NN	O	O
of	NN	O	O
109	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
detected	NN	O	O
among	NN	O	O
181	NN	O	O
patients	NN	O	O
analyzed	NN	O	O
with	NN	O	O
the	NN	O	O
entire	NN	O	O
dystrophin	NN	O	O
cDNA	NN	O	O
.	NN	O	O

Endpoints	NN	O	O
for	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
deletions	NN	O	O
were	NN	O	O
further	NN	O	O
characterized	NN	O	O
using	NN	O	O
two	NN	O	O
genomic	NN	O	O
probes	NN	O	O
,	NN	O	O
p20	NN	O	O
(	NN	O	O
DXS269	NN	O	O
;	NN	O	O
Wapenaar	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
)	NN	O	O
and	NN	O	O
GMGX11	NN	O	O
(	NN	O	O
DXS239	NN	O	O
;	NN	O	O
present	NN	O	O
paper	NN	O	O
)	NN	O	O
.	NN	O	O

Clinical	NN	O	O
findings	NN	O	O
are	NN	O	O
presented	NN	O	O
for	NN	O	O
all	NN	O	O
80	NN	O	O
patients	NN	O	O
allowing	NN	O	O
a	NN	O	O
correlation	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
severity	NN	O	O
with	NN	O	O
the	NN	O	O
genotype	NN	O	O
.	NN	O	O

Thirty	NN	O	O
-	NN	O	O
eight	NN	O	O
independent	NN	O	O
patients	NN	O	O
were	NN	O	O
old	NN	O	O
enough	NN	O	O
to	NN	O	O
be	NN	O	O
classified	NN	O	O
as	NN	O	O
DMD	NN	O	B-Disease
,	NN	O	O
BMD	NN	O	B-Disease
,	NN	O	O
or	NN	O	O
intermediate	NN	O	O
phenotype	NN	O	O
and	NN	O	O
had	NN	O	O
deletions	NN	O	O
of	NN	O	O
exons	NN	O	O
with	NN	O	O
sequenced	NN	O	O
intron	NN	O	O
/	NN	O	O
exon	NN	O	O
boundaries	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
eight	NN	O	O
BMD	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
one	NN	O	O
intermediate	NN	O	O
patient	NN	O	O
had	NN	O	O
gene	NN	O	O
deletions	NN	O	O
predicted	NN	O	O
to	NN	O	O
leave	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
intact	NN	O	O
,	NN	O	O
while	NN	O	O
21	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
7	NN	O	O
intermediate	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
BMD	NN	O	B-Disease
patient	NN	O	O
had	NN	O	O
gene	NN	O	O
deletions	NN	O	O
predicted	NN	O	O
to	NN	O	O
disrupt	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
with	NN	O	O
two	NN	O	O
exceptions	NN	O	O
,	NN	O	O
frameshift	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
resulted	NN	O	O
in	NN	O	O
more	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
than	NN	O	O
did	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
deletions	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
recent	NN	O	O
findings	NN	O	O
by	NN	O	O
Baumbach	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
and	NN	O	O
Koenig	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
but	NN	O	O
is	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
findings	NN	O	O
,	NN	O	O
by	NN	O	O
Malhotra	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
at	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Cloning	NN	O	O
of	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
a	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
identifies	NN	O	O
the	NN	O	O
AN2	NN	O	O
locus	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
translocations	NN	O	O
involving	NN	O	O
11p13	NN	O	O
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
aniridia	NN	O	I-Disease
in	NN	O	O
two	NN	O	O
kindreds	NN	O	O
highlighting	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
AN2	NN	O	O
locus	NN	O	O
.	NN	O	O

This	NN	O	O
locus	NN	O	O
is	NN	O	O
also	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
WAGR	NN	O	B-Disease
complex	NN	O	O
(	NN	O	O
Wilms	NN	O	B-Disease
tumor	NN	O	I-Disease
,	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
genitourinary	NN	O	B-Disease
abnormalities	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
kindred	NN	O	O
,	NN	O	O
the	NN	O	O
translocation	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
deletion	NN	O	O
,	NN	O	O
and	NN	O	O
probes	NN	O	O
for	NN	O	O
this	NN	O	O
region	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
and	NN	O	O
clone	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
the	NN	O	O
translocation	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
kindred	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
phage	NN	O	O
restriction	NN	O	O
maps	NN	O	O
exclude	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
any	NN	O	O
sizable	NN	O	O
deletion	NN	O	O
in	NN	O	O
this	NN	O	O
case	NN	O	O
.	NN	O	O

Sequences	NN	O	O
at	NN	O	O
the	NN	O	O
chromosome	NN	O	O
11	NN	O	O
breakpoint	NN	O	O
are	NN	O	O
conserved	NN	O	O
in	NN	O	O
multiple	NN	O	O
species	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
translocation	NN	O	O
falls	NN	O	O
within	NN	O	O
the	NN	O	O
AN2	NN	O	O
gene	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
C2	NN	O	O
gene	NN	O	O
and	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
on	NN	O	O
human	NN	O	O
chromosome	NN	O	O
19	NN	O	O
reveals	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
in	NN	O	O
a	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
human	NN	O	O
apolipoprotein	NN	O	O
C2	NN	O	O
(	NN	O	O
APOC2	NN	O	O
)	NN	O	O
,	NN	O	O
situated	NN	O	O
on	NN	O	O
the	NN	O	O
proximal	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
19	NN	O	O
,	NN	O	O
is	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
form	NN	O	O
of	NN	O	O
adult	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Six	NN	O	O
APOC2	NN	O	O
RFLPs	NN	O	O
(	NN	O	O
TaqI	NN	O	O
,	NN	O	O
BglI	NN	O	O
,	NN	O	O
BanI	NN	O	O
,	NN	O	O
BamHI	NN	O	O
,	NN	O	O
NcoI	NN	O	O
,	NN	O	O
and	NN	O	O
AvaII	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
to	NN	O	O
date	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
conducted	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
DM	NN	O	B-Disease
linkage	NN	O	O
study	NN	O	O
utilizing	NN	O	O
all	NN	O	O
six	NN	O	O
RFLPs	NN	O	O
and	NN	O	O
involving	NN	O	O
50	NN	O	O
families	NN	O	O
and	NN	O	O
372	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
informative	NN	O	O
RFLPs	NN	O	O
are	NN	O	O
,	NN	O	O
in	NN	O	O
descending	NN	O	O
order	NN	O	O
,	NN	O	O
NcoI	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
64	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
)	NN	O	O
,	NN	O	O
BglI	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
12	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
)	NN	O	O
,	NN	O	O
AvaII	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
02	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
03	NN	O	O
)	NN	O	O
,	NN	O	O
BanI	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
5	NN	O	O
.	NN	O	O
76	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
04	NN	O	O
)	NN	O	O
,	NN	O	O
TaqI	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
4	NN	O	O
.	NN	O	O
29	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
06	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
BamHI	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
1	NN	O	O
.	NN	O	O
75	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
01	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
substantial	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
lod	NN	O	O
scores	NN	O	O
over	NN	O	O
those	NN	O	O
seen	NN	O	O
with	NN	O	O
the	NN	O	O
individual	NN	O	O
RFLPs	NN	O	O
was	NN	O	O
obtained	NN	O	O
when	NN	O	O
the	NN	O	O
linkage	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
APOC2	NN	O	O
haplotype	NN	O	O
(	NN	O	O
composed	NN	O	O
of	NN	O	O
the	NN	O	O
six	NN	O	O
RFLPs	NN	O	O
)	NN	O	O
was	NN	O	O
studied	NN	O	O
(	NN	O	O
lod	NN	O	O
=	NN	O	O
17	NN	O	O
.	NN	O	O
87	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
04	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
observed	NN	O	O
significant	NN	O	O
inter	NN	O	O
-	NN	O	O
APOC2	NN	O	O
RFLP	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
the	NN	O	O
three	NN	O	O
most	NN	O	O
informative	NN	O	O
RFLPs	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
BanI	NN	O	O
,	NN	O	O
TaqI	NN	O	O
,	NN	O	O
and	NN	O	O
either	NN	O	O
BglI	NN	O	O
,	NN	O	O
AvaII	NN	O	O
,	NN	O	O
or	NN	O	O
NcoI	NN	O	O
polymorphisms	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
between	NN	O	O
DM	NN	O	O
and	NN	O	O
APOC2	NN	O	O
in	NN	O	O
our	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
population	NN	O	O
(	NN	O	O
standardized	NN	O	O
disequilibrium	NN	O	O
constant	NN	O	O
phi	NN	O	O
=	NN	O	O
.	NN	O	O
22	NN	O	O
,	NN	O	O
chi	NN	O	O
2	NN	O	O
=	NN	O	O
5	NN	O	O
.	NN	O	O
12	NN	O	O
,	NN	O	O
df	NN	O	O
=	NN	O	O
1	NN	O	O
,	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0	NN	O	O
.	NN	O	O
04	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
between	NN	O	O
APOC2	NN	O	O
and	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

Phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
Polynesians	NN	O	O
:	NN	O	O
evolutionary	NN	O	O
origins	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
alleles	NN	O	O
associated	NN	O	O
with	NN	O	O
severe	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
630	NN	O	O
haplotypes	NN	O	O
for	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
gene	NN	O	O
locus	NN	O	O
were	NN	O	O
established	NN	O	O
in	NN	O	O
five	NN	O	O
groups	NN	O	O
of	NN	O	O
Polynesians	NN	O	O
comprising	NN	O	O
Samoans	NN	O	O
,	NN	O	O
Tongans	NN	O	O
,	NN	O	O
Cook	NN	O	O
Islanders	NN	O	O
,	NN	O	O
Maori	NN	O	O
,	NN	O	O
and	NN	O	O
Niueans	NN	O	O
.	NN	O	O

Considerable	NN	O	O
genetic	NN	O	O
continuity	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
between	NN	O	O
these	NN	O	O
widely	NN	O	O
dispersed	NN	O	O
populations	NN	O	O
,	NN	O	O
since	NN	O	O
three	NN	O	O
common	NN	O	O
haplotypes	NN	O	O
(	NN	O	O
4	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
)	NN	O	O
constituted	NN	O	O
over	NN	O	O
95	NN	O	O
%	NN	O	O
of	NN	O	O
alleles	NN	O	O
.	NN	O	O

A	NN	O	O
control	NN	O	O
group	NN	O	O
of	NN	O	O
individuals	NN	O	O
from	NN	O	O
Southeast	NN	O	O
Asia	NN	O	O
shared	NN	O	O
the	NN	O	O
same	NN	O	O
major	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
,	NN	O	O
with	NN	O	O
Polynesians	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
further	NN	O	O
support	NN	O	O
for	NN	O	O
the	NN	O	O
theories	NN	O	O
of	NN	O	O
genetic	NN	O	O
homogeneity	NN	O	O
and	NN	O	O
of	NN	O	O
Asian	NN	O	O
affinities	NN	O	O
of	NN	O	O
the	NN	O	O
Polynesian	NN	O	O
precursor	NN	O	O
populations	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
severe	NN	O	O
phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
both	NN	O	O
Polynesians	NN	O	O
and	NN	O	O
Southeast	NN	O	O
Asians	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
PAH	NN	O	O
haplotypes	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
,	NN	O	O
on	NN	O	O
which	NN	O	O
the	NN	O	O
severe	NN	O	O
PKU	NN	O	B-Disease
mutants	NN	O	O
have	NN	O	O
arisen	NN	O	O
among	NN	O	O
Caucasians	NN	O	O
.	NN	O	O
.	NN	O	O

Preferential	NN	O	O
germline	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
paternal	NN	O	O
allele	NN	O	O
in	NN	O	O
retinoblastoma	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
event	NN	O	O
triggering	NN	O	O
malignant	NN	O	O
proliferation	NN	O	O
in	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-Disease
tumours	NN	O	I-Disease
is	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
for	NN	O	O
chromosome	NN	O	O
13q14	NN	O	O
,	NN	O	O
whereby	NN	O	O
the	NN	O	O
normal	NN	O	O
retinoblastoma	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
RB1	NN	O	O
)	NN	O	O
allele	NN	O	O
is	NN	O	O
lost	NN	O	O
and	NN	O	O
an	NN	O	O
already	NN	O	O
mutated	NN	O	O
RB1	NN	O	O
allele	NN	O	O
remains	NN	O	O
in	NN	O	O
the	NN	O	O
tumour	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
first	NN	O	O
allele	NN	O	O
suffers	NN	O	O
a	NN	O	O
mutational	NN	O	O
event	NN	O	O
-	NN	O	O
-	NN	O	O
deletion	NN	O	O
,	NN	O	O
duplication	NN	O	O
or	NN	O	O
point	NN	O	O
mutation	NN	O	O
(	NN	O	O
manuscript	NN	O	O
in	NN	O	O
preparation	NN	O	O
)	NN	O	O
-	NN	O	O
-	NN	O	O
either	NN	O	O
in	NN	O	O
the	NN	O	O
germ	NN	O	O
line	NN	O	O
(	NN	O	O
all	NN	O	O
bilateral	NN	O	O
patients	NN	O	O
)	NN	O	O
or	NN	O	O
in	NN	O	O
a	NN	O	O
somatic	NN	O	O
retinal	NN	O	O
cell	NN	O	O
(	NN	O	O
most	NN	O	O
unilateral	NN	O	O
patients	NN	O	O
)	NN	O	O
.	NN	O	O

Most	NN	O	O
bilateral	NN	O	O
patients	NN	O	O
have	NN	O	O
no	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-Disease
and	NN	O	O
are	NN	O	O
presumed	NN	O	O
to	NN	O	O
have	NN	O	O
new	NN	O	O
germline	NN	O	O
mutations	NN	O	O
which	NN	O	O
arose	NN	O	O
in	NN	O	O
the	NN	O	O
egg	NN	O	O
,	NN	O	O
sperm	NN	O	O
or	NN	O	O
early	NN	O	O
embryo	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
parental	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
retained	NN	O	O
allele	NN	O	O
in	NN	O	O
nine	NN	O	O
retinoblastoma	NN	O	B-Disease
tumours	NN	O	I-Disease
from	NN	O	O
eight	NN	O	O
unrelated	NN	O	O
non	NN	O	O
-	NN	O	O
familial	NN	O	O
cases	NN	O	O
by	NN	O	O
using	NN	O	O
RB1	NN	O	O
-	NN	O	O
linked	NN	O	O
genetic	NN	O	O
markers	NN	O	O
.	NN	O	O

Six	NN	O	O
tumours	NN	O	B-Disease
retained	NN	O	O
the	NN	O	O
paternal	NN	O	O
allele	NN	O	O
and	NN	O	O
three	NN	O	O
retained	NN	O	O
the	NN	O	O
maternal	NN	O	O
allele	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
three	NN	O	O
unilateral	NN	O	B-Disease
tumours	NN	O	I-Disease
,	NN	O	O
only	NN	O	O
one	NN	O	O
retained	NN	O	O
the	NN	O	O
paternal	NN	O	O
RB1	NN	O	O
allele	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
paternal	NN	O	O
RB1	NN	O	O
allele	NN	O	O
is	NN	O	O
preferentially	NN	O	O
retained	NN	O	O
in	NN	O	O
retinoblastoma	NN	O	B-Disease
,	NN	O	O
as	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
case	NN	O	O
in	NN	O	O
osteosarcoma	NN	O	B-Disease
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
tumours	NN	O	B-Disease
from	NN	O	O
four	NN	O	O
of	NN	O	O
the	NN	O	O
five	NN	O	O
bilateral	NN	O	O
patients	NN	O	O
retained	NN	O	O
the	NN	O	O
paternal	NN	O	O
RB1	NN	O	O
allele	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
either	NN	O	O
that	NN	O	O
new	NN	O	O
germline	NN	O	O
RB1	NN	O	O
mutations	NN	O	O
arise	NN	O	O
more	NN	O	O
frequently	NN	O	O
during	NN	O	O
spermatogenesis	NN	O	O
than	NN	O	O
during	NN	O	O
oogenesis	NN	O	O
,	NN	O	O
or	NN	O	O
that	NN	O	O
imprinting	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
embryo	NN	O	O
affects	NN	O	O
chromosomal	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
mutation	NN	O	O
.	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
in	NN	O	O
Turkish	NN	O	O
phenylketonuria	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
estimated	NN	O	O
the	NN	O	O
haplotype	NN	O	O
distribution	NN	O	O
of	NN	O	O
mutant	NN	O	O
and	NN	O	O
normal	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
alleles	NN	O	O
for	NN	O	O
17	NN	O	O
Turkish	NN	O	O
phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
families	NN	O	O
20	NN	O	O
normal	NN	O	O
and	NN	O	O
27	NN	O	O
mutated	NN	O	O
PAH	NN	O	O
alleles	NN	O	O
could	NN	O	O
be	NN	O	O
identified	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
latter	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
prevalent	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
haplotype	NN	O	O
6	NN	O	O
(	NN	O	O
29	NN	O	O
.	NN	O	O
6	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
1	NN	O	O
(	NN	O	O
18	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
36	NN	O	O
(	NN	O	O
11	NN	O	O
.	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
normal	NN	O	O
alleles	NN	O	O
were	NN	O	O
preferentially	NN	O	O
associated	NN	O	O
with	NN	O	O
haplotype	NN	O	O
1	NN	O	O
(	NN	O	O
20	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
19	NN	O	O
different	NN	O	O
haplotypes	NN	O	O
observed	NN	O	O
,	NN	O	O
5	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
described	NN	O	O
previously	NN	O	O
.	NN	O	O

The	NN	O	O
haplotype	NN	O	O
distribution	NN	O	O
differed	NN	O	O
significantly	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
Northern	NN	O	O
European	NN	O	O
population	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
polymorphic	NN	O	O
sites	NN	O	O
were	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
PKU	NN	O	B-Disease
.	NN	O	O

No	NN	O	O
deletions	NN	O	O
of	NN	O	O
exon	NN	O	O
sequences	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
families	NN	O	O
analysed	NN	O	O
.	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
locus	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
63	NN	O	O
families	NN	O	O
with	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
linkage	NN	O	O
between	NN	O	O
HD	NN	O	B-Disease
and	NN	O	O
G8	NN	O	O
(	NN	O	O
D4S10	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
families	NN	O	O
included	NN	O	O
57	NN	O	O
Caucasian	NN	O	O
,	NN	O	O
four	NN	O	O
Black	NN	O	O
American	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
Japanese	NN	O	O
.	NN	O	O

The	NN	O	O
combined	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
was	NN	O	O
87	NN	O	O
.	NN	O	O

69	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

04	NN	O	O
(	NN	O	O
99	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
0	NN	O	O
.	NN	O	O
018	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
071	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
maximum	NN	O	O
frequency	NN	O	O
of	NN	O	O
recombination	NN	O	O
was	NN	O	O
0	NN	O	O
.	NN	O	O

03	NN	O	O
in	NN	O	O
males	NN	O	O
and	NN	O	O
0	NN	O	O
.	NN	O	O

05	NN	O	O
in	NN	O	O
females	NN	O	O
.	NN	O	O

Fifty	NN	O	O
-	NN	O	O
seven	NN	O	O
families	NN	O	O
gave	NN	O	O
positive	NN	O	O
lod	NN	O	O
scores	NN	O	O
;	NN	O	O
five	NN	O	O
small	NN	O	O
families	NN	O	O
gave	NN	O	O
mildly	NN	O	O
negative	NN	O	O
lod	NN	O	O
scores	NN	O	O
.	NN	O	O

The	NN	O	O
maximum	NN	O	O
likelihood	NN	O	O
estimate	NN	O	O
of	NN	O	O
alpha	NN	O	O
,	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
linked	NN	O	O
loci	NN	O	O
,	NN	O	O
was	NN	O	O
1	NN	O	O
.	NN	O	O

0	NN	O	O
with	NN	O	O
a	NN	O	O
lower	NN	O	O
99	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

88	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
only	NN	O	O
one	NN	O	O
HD	NN	O	B-Disease
locus	NN	O	O
,	NN	O	O
although	NN	O	O
a	NN	O	O
second	NN	O	O
rare	NN	O	O
locus	NN	O	O
cannot	NN	O	O
be	NN	O	O
ruled	NN	O	O
out	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
and	NN	O	O
multipoint	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
Finnish	NN	O	O
choroideremia	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

Multipoint	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
choroideremia	NN	O	B-Disease
(	NN	O	O
TCD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
seven	NN	O	O
X	NN	O	O
chromosomal	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
RFLPs	NN	O	O
)	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
in	NN	O	O
18	NN	O	O
Finnish	NN	O	O
TCD	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
place	NN	O	O
TCD	NN	O	B-Disease
distal	NN	O	O
to	NN	O	O
PGK	NN	O	O
and	NN	O	O
DXS72	NN	O	O
,	NN	O	O
very	NN	O	O
close	NN	O	O
to	NN	O	O
DXYS1	NN	O	O
and	NN	O	O
DXYS5	NN	O	O
(	NN	O	O
Zmax	NN	O	O
=	NN	O	O
24	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
)	NN	O	O
and	NN	O	O
proximal	NN	O	O
to	NN	O	O
DXYS4	NN	O	O
and	NN	O	O
DXYS12	NN	O	O
.	NN	O	O

This	NN	O	O
agrees	NN	O	O
with	NN	O	O
the	NN	O	O
data	NN	O	O
obtained	NN	O	O
from	NN	O	O
other	NN	O	O
linkage	NN	O	O
studies	NN	O	O
and	NN	O	O
from	NN	O	O
physical	NN	O	O
mapping	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
TCD	NN	O	B-Disease
males	NN	O	O
and	NN	O	O
carrier	NN	O	O
females	NN	O	O
studied	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
DXYS1	NN	O	O
allele	NN	O	O
in	NN	O	O
coupling	NN	O	O
with	NN	O	O
TCD	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
Northeastern	NN	O	O
Finland	NN	O	O
,	NN	O	O
66	NN	O	O
/	NN	O	O
69	NN	O	O
chromosomes	NN	O	O
carrying	NN	O	O
TCD	NN	O	B-Disease
had	NN	O	O
the	NN	O	O
same	NN	O	O
haplotype	NN	O	O
at	NN	O	O
loci	NN	O	O
DXS72	NN	O	O
,	NN	O	O
DXYS1	NN	O	O
,	NN	O	O
DXYS4	NN	O	O
,	NN	O	O
and	NN	O	O
DXYS12	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
haplotype	NN	O	O
is	NN	O	O
seen	NN	O	O
in	NN	O	O
only	NN	O	O
15	NN	O	O
/	NN	O	O
99	NN	O	O
chromosomes	NN	O	O
not	NN	O	O
carrying	NN	O	O
TCD	NN	O	B-Disease
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
71	NN	O	O
/	NN	O	O
104	NN	O	O
non	NN	O	O
-	NN	O	O
TCD	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
the	NN	O	O
haplotype	NN	O	O
at	NN	O	O
six	NN	O	O
marker	NN	O	O
loci	NN	O	O
is	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
seen	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
76	NN	O	O
TCD	NN	O	B-Disease
chromosomes	NN	O	O
.	NN	O	O

This	NN	O	O
supports	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
large	NN	O	O
Northern	NN	O	O
Finnish	NN	O	O
choroideremia	NN	O	B-Disease
pedigrees	NN	O	O
,	NN	O	O
comprising	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
over	NN	O	O
80	NN	O	O
living	NN	O	O
patients	NN	O	O
representing	NN	O	O
more	NN	O	O
than	NN	O	O
a	NN	O	O
fifth	NN	O	O
of	NN	O	O
all	NN	O	O
TCD	NN	O	B-Disease
patients	NN	O	O
described	NN	O	O
worldwide	NN	O	O
,	NN	O	O
carry	NN	O	O
the	NN	O	O
same	NN	O	O
mutation	NN	O	O
.	NN	O	O

These	NN	O	O
linkage	NN	O	O
and	NN	O	O
haplotype	NN	O	O
data	NN	O	O
provide	NN	O	O
improved	NN	O	O
opportunities	NN	O	O
for	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
based	NN	O	O
on	NN	O	O
RFLP	NN	O	O
studies	NN	O	O
.	NN	O	O
.	NN	O	O

Autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
linked	NN	O	O
to	NN	O	O
the	NN	O	O
chromosome	NN	O	O
11p13	NN	O	O
markers	NN	O	O
catalase	NN	O	O
and	NN	O	O
D11S151	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
Dutch	NN	O	O
family	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
large	NN	O	O
pedigree	NN	O	O
with	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
found	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
aniridia	NN	O	B-Disease
locus	NN	O	O
AN2	NN	O	O
and	NN	O	O
the	NN	O	O
markers	NN	O	O
catalase	NN	O	O
(	NN	O	O
CAT	NN	O	O
)	NN	O	O
(	NN	O	O
zeta	NN	O	O
=	NN	O	O
7	NN	O	O
.	NN	O	O
27	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
00	NN	O	O
)	NN	O	O
and	NN	O	O
D11S151	NN	O	O
(	NN	O	O
zeta	NN	O	O
=	NN	O	O
3	NN	O	O
.	NN	O	O
86	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
10	NN	O	O
)	NN	O	O
flanking	NN	O	O
the	NN	O	O
AN2	NN	O	O
locus	NN	O	O
on	NN	O	O
11p13	NN	O	O
.	NN	O	O

Positive	NN	O	O
lod	NN	O	O
scores	NN	O	O
were	NN	O	O
also	NN	O	O
obtained	NN	O	O
for	NN	O	O
the	NN	O	O
11p13	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
11p14	NN	O	O
markers	NN	O	O
D11S16	NN	O	O
and	NN	O	O
FSHB	NN	O	O
with	NN	O	O
the	NN	O	O
linkage	NN	O	O
group	NN	O	O
CAT	NN	O	O
/	NN	O	O
AN2	NN	O	O
/	NN	O	O
D11S151	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
aniridia	NN	O	B-Disease
in	NN	O	O
this	NN	O	O
family	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
AN2	NN	O	O
locus	NN	O	O
on	NN	O	O
11p13	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
excluded	NN	O	O
linkage	NN	O	O
(	NN	O	O
zeta	NN	O	O
less	NN	O	O
than	NN	O	O
-	NN	O	O
2	NN	O	O
at	NN	O	O
theta	NN	O	O
less	NN	O	O
than	NN	O	O
0	NN	O	O
.	NN	O	O
18	NN	O	O
)	NN	O	O
between	NN	O	O
the	NN	O	O
aniridia	NN	O	B-Disease
and	NN	O	O
the	NN	O	O
chromosome	NN	O	O
2p25	NN	O	O
marker	NN	O	O
D2S1	NN	O	O
(	NN	O	O
linked	NN	O	O
to	NN	O	O
ACP1	NN	O	O
)	NN	O	O
.	NN	O	O

Recombination	NN	O	O
events	NN	O	O
that	NN	O	O
locate	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
distal	NN	O	O
to	NN	O	O
APOC2	NN	O	O
on	NN	O	O
19q	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
a	NN	O	O
recombination	NN	O	O
in	NN	O	O
an	NN	O	O
individual	NN	O	O
with	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
which	NN	O	O
placed	NN	O	O
the	NN	O	O
markers	NN	O	O
D19S19	NN	O	O
and	NN	O	O
APOC2	NN	O	O
on	NN	O	O
the	NN	O	O
same	NN	O	O
side	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

Haplotyping	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
with	NN	O	O
more	NN	O	O
recently	NN	O	O
characterized	NN	O	O
probes	NN	O	O
which	NN	O	O
are	NN	O	O
either	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
DM	NN	O	B-Disease
or	NN	O	O
distal	NN	O	O
to	NN	O	O
the	NN	O	O
linkage	NN	O	O
group	NN	O	O
at	NN	O	O
q13	NN	O	O
.	NN	O	O

2	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
locus	NN	O	O
is	NN	O	O
distal	NN	O	O
to	NN	O	O
APOC2	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
confirmed	NN	O	O
by	NN	O	O
other	NN	O	O
recombinants	NN	O	O
where	NN	O	O
DM	NN	O	B-Disease
segregates	NN	O	O
with	NN	O	O
distal	NN	O	O
probes	NN	O	O
.	NN	O	O

Additional	NN	O	O
marker	NN	O	O
to	NN	O	O
marker	NN	O	O
recombinations	NN	O	O
in	NN	O	O
unaffected	NN	O	O
individuals	NN	O	O
are	NN	O	O
reported	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
order	NN	O	O
and	NN	O	O
orientation	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
linkage	NN	O	O
group	NN	O	O
as	NN	O	O
pter	NN	O	O
-	NN	O	O
(	NN	O	O
INSR	NN	O	O
,	NN	O	O
LDLR	NN	O	O
,	NN	O	O
S9	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
S19	NN	O	O
,	NN	O	O
BCL3	NN	O	O
,	NN	O	O
APOC2	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
CKMM	NN	O	O
,	NN	O	O
DM	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
S22	NN	O	O
,	NN	O	O
+	NN	O	O
+	NN	O	O
+	NN	O	O
PRKCG	NN	O	O
)	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
cannot	NN	O	O
determine	NN	O	O
whether	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
proximal	NN	O	O
or	NN	O	O
distal	NN	O	O
to	NN	O	O
CKMM	NN	O	O
.	NN	O	O

The	NN	O	O
consequences	NN	O	O
of	NN	O	O
this	NN	O	O
probe	NN	O	O
order	NN	O	O
for	NN	O	O
antenatal	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
future	NN	O	O
research	NN	O	O
aiming	NN	O	O
to	NN	O	O
isolate	NN	O	O
the	NN	O	O
gene	NN	O	O
which	NN	O	O
is	NN	O	O
affected	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
and	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
transcription	NN	O	O
.	NN	O	O

Inactivation	NN	O	O
of	NN	O	O
both	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
during	NN	O	O
normal	NN	O	O
retinal	NN	O	O
development	NN	O	O
initiates	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
retinoblastoma	NN	O	B-Disease
(	NN	O	I-Disease
RB	NN	O	I-Disease
)	NN	O	I-Disease
tumor	NN	O	I-Disease
.	NN	O	O

To	NN	O	O
identify	NN	O	O
the	NN	O	O
mutations	NN	O	O
which	NN	O	O
inactivate	NN	O	O
RB1	NN	O	O
,	NN	O	O
21	NN	O	O
RB	NN	O	B-Disease
tumors	NN	O	I-Disease
isolated	NN	O	O
from	NN	O	O
19	NN	O	O
patients	NN	O	O
were	NN	O	O
analyzed	NN	O	O
with	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
or	NN	O	O
an	NN	O	O
RNase	NN	O	O
protection	NN	O	O
assay	NN	O	O
or	NN	O	O
both	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
13	NN	O	O
of	NN	O	O
21	NN	O	O
RB	NN	O	B-Disease
tumors	NN	O	I-Disease
;	NN	O	O
in	NN	O	O
8	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
precise	NN	O	O
errors	NN	O	O
in	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
were	NN	O	O
characterized	NN	O	O
.	NN	O	O

Each	NN	O	O
of	NN	O	O
four	NN	O	O
germ	NN	O	O
line	NN	O	O
mutations	NN	O	O
involved	NN	O	O
a	NN	O	O
small	NN	O	O
deletion	NN	O	O
or	NN	O	O
duplication	NN	O	O
,	NN	O	O
while	NN	O	O
three	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
were	NN	O	O
point	NN	O	O
mutations	NN	O	O
leading	NN	O	O
to	NN	O	O
splice	NN	O	O
alterations	NN	O	O
and	NN	O	O
loss	NN	O	O
of	NN	O	O
an	NN	O	O
exon	NN	O	O
from	NN	O	O
the	NN	O	O
mature	NN	O	O
RB1	NN	O	O
mRNA	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
detect	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
in	NN	O	O
lymphoblasts	NN	O	O
of	NN	O	O
three	NN	O	O
bilaterally	NN	O	O
affected	NN	O	O
patients	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
mutation	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
and	NN	O	O
transcripts	NN	O	O
containing	NN	O	O
the	NN	O	O
mutations	NN	O	O
were	NN	O	O
obvious	NN	O	O
in	NN	O	O
the	NN	O	O
RB	NN	O	B-Disease
tumors	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
normal	NN	O	O
RB1	NN	O	O
allele	NN	O	O
.	NN	O	O

The	NN	O	O
variations	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
mutant	NN	O	O
transcripts	NN	O	O
suggest	NN	O	O
deregulation	NN	O	O
of	NN	O	O
RB1	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
RB1	NN	O	O
gene	NN	O	O
product	NN	O	O
.	NN	O	O
.	NN	O	O

Partial	NN	O	O
deletion	NN	O	O
8q	NN	O	O
without	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
recognisable	NN	O	O
syndrome	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
two	NN	O	O
de	NN	O	O
novo	NN	O	O
cases	NN	O	O
of	NN	O	O
del	NN	O	O
(	NN	O	O
8	NN	O	O
)	NN	O	O
(	NN	O	O
pter	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
q24	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
with	NN	O	O
breakpoints	NN	O	O
involving	NN	O	O
the	NN	O	O
distal	NN	O	O
part	NN	O	O
of	NN	O	O
band	NN	O	O
8q24	NN	O	O
.	NN	O	O

1	NN	O	O
1	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
features	NN	O	O
were	NN	O	O
similar	NN	O	O
and	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
obvious	NN	O	O
stigmata	NN	O	O
of	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
LGS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
three	NN	O	O
other	NN	O	O
cases	NN	O	O
reported	NN	O	O
with	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
chromosome	NN	O	O
8	NN	O	O
at	NN	O	O
approximately	NN	O	O
the	NN	O	O
same	NN	O	O
breakpoint	NN	O	O
,	NN	O	O
one	NN	O	O
without	NN	O	O
LGS	NN	O	B-Disease
and	NN	O	O
some	NN	O	O
similarities	NN	O	O
to	NN	O	O
our	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
other	NN	O	O
two	NN	O	O
with	NN	O	O
LGS	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
would	NN	O	O
support	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
the	NN	O	O
critical	NN	O	O
segment	NN	O	O
for	NN	O	O
the	NN	O	O
assignment	NN	O	O
of	NN	O	O
LGS	NN	O	B-Disease
is	NN	O	O
proximal	NN	O	O
to	NN	O	O
or	NN	O	O
involves	NN	O	O
the	NN	O	O
proximal	NN	O	O
part	NN	O	O
of	NN	O	O
8q24	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
but	NN	O	O
a	NN	O	O
review	NN	O	O
of	NN	O	O
published	NN	O	O
reports	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
aetiology	NN	O	O
of	NN	O	O
LGS	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
a	NN	O	O
more	NN	O	O
complex	NN	O	O
issue	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
is	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
muscle	NN	O	O
-	NN	O	O
type	NN	O	O
creatine	NN	O	O
kinase	NN	O	O
(	NN	O	O
CKMM	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
genetic	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
creatine	NN	O	O
kinase	NN	O	O
muscle	NN	O	O
type	NN	O	O
(	NN	O	O
CKMM	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
65	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
families	NN	O	O
from	NN	O	O
Canada	NN	O	O
and	NN	O	O
the	NN	O	O
Netherlands	NN	O	O
,	NN	O	O
a	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
(	NN	O	O
Zmax	NN	O	O
)	NN	O	O
of	NN	O	O
22	NN	O	O
.	NN	O	O

8	NN	O	O
at	NN	O	O
a	NN	O	O
recombination	NN	O	O
frequency	NN	O	O
(	NN	O	O
theta	NN	O	O
)	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

03	NN	O	O
was	NN	O	O
obtained	NN	O	O
.	NN	O	O

Tight	NN	O	O
linkage	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
for	NN	O	O
CKMM	NN	O	O
and	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
apolipoprotein	NN	O	O
C2	NN	O	O
(	NN	O	O
ApoC2	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
establishes	NN	O	O
CKMM	NN	O	O
as	NN	O	O
a	NN	O	O
useful	NN	O	O
marker	NN	O	O
for	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease

Color	NN	O	B-Disease
vision	NN	O	I-Disease
defects	NN	O	I-Disease
in	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
abnormal	NN	O	B-Disease
color	NN	O	I-Disease
vision	NN	O	I-Disease
and	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
27	NN	O	O
AMN	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
31	NN	O	O
age	NN	O	O
-	NN	O	O
matched	NN	O	O
controls	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
Farnsworth	NN	O	O
-	NN	O	O
Munsell	NN	O	O
100	NN	O	O
Hue	NN	O	O
test	NN	O	O
.	NN	O	O

Twelve	NN	O	O
(	NN	O	O
44	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
27	NN	O	O
patients	NN	O	O
showed	NN	O	O
test	NN	O	O
scores	NN	O	O
significantly	NN	O	O
above	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
axes	NN	O	O
of	NN	O	O
bipolarity	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
testing	NN	O	O
differed	NN	O	O
widely	NN	O	O
between	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
abnormal	NN	O	O
scores	NN	O	O
,	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
different	NN	O	O
alterations	NN	O	O
in	NN	O	O
visual	NN	O	O
pigment	NN	O	O
genes	NN	O	O
occur	NN	O	O
in	NN	O	O
different	NN	O	O
AMN	NN	O	B-Disease
kindreds	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
confirm	NN	O	O
our	NN	O	O
earlier	NN	O	O
impression	NN	O	O
that	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
abnormal	NN	O	B-Disease
color	NN	O	I-Disease
vision	NN	O	I-Disease
is	NN	O	O
increased	NN	O	O
in	NN	O	O
these	NN	O	O
kindreds	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
supports	NN	O	O
our	NN	O	O
contentions	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
AMN	NN	O	B-Disease
(	NN	O	O
and	NN	O	O
its	NN	O	O
companion	NN	O	O
,	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
very	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
visual	NN	O	O
pigment	NN	O	O
loci	NN	O	O
at	NN	O	O
Xq28	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
this	NN	O	O
proximity	NN	O	O
might	NN	O	O
provide	NN	O	O
the	NN	O	O
opportunity	NN	O	O
to	NN	O	O
observe	NN	O	O
contiguous	NN	O	B-Disease
gene	NN	O	I-Disease
defects	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
female	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patient	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
female	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
recessive	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
hypoxanthine	NN	O	B-Disease
phosphoribosyltransferase	NN	O	I-Disease
[	NN	O	I-Disease
HPRT	NN	O	I-Disease
]	NN	O	I-Disease
deficiency	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

Cytogenetic	NN	O	O
and	NN	O	O
carrier	NN	O	O
studies	NN	O	O
revealed	NN	O	O
structurally	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
for	NN	O	O
this	NN	O	O
patient	NN	O	O
and	NN	O	O
her	NN	O	O
parents	NN	O	O
and	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
arose	NN	O	O
through	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
gametic	NN	O	O
event	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
this	NN	O	O
patients	NN	O	O
DNA	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
her	NN	O	O
parents	NN	O	O
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
microdeletion	NN	O	O
,	NN	O	O
which	NN	O	O
occurred	NN	O	O
within	NN	O	O
a	NN	O	O
maternal	NN	O	O
gamete	NN	O	O
and	NN	O	O
involved	NN	O	O
the	NN	O	O
entire	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
,	NN	O	O
was	NN	O	O
partially	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
disease	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O

Somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
,	NN	O	O
generated	NN	O	O
to	NN	O	O
separate	NN	O	O
maternal	NN	O	O
and	NN	O	O
paternal	NN	O	O
X	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
showed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
additional	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
enzymes	NN	O	O
,	NN	O	O
phosphoglycerate	NN	O	O
kinase	NN	O	O
and	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
,	NN	O	O
were	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
contained	NN	O	O
the	NN	O	O
maternal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
functionally	NN	O	O
inactive	NN	O	O
paternal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
comparison	NN	O	O
of	NN	O	O
methylation	NN	O	O
patterns	NN	O	O
within	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
gene	NN	O	O
regulation	NN	O	O
revealed	NN	O	O
differences	NN	O	O
between	NN	O	O
DNA	NN	O	O
from	NN	O	O
the	NN	O	O
father	NN	O	O
and	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
in	NN	O	O
keeping	NN	O	O
with	NN	O	O
an	NN	O	O
active	NN	O	O
HPRT	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
father	NN	O	O
and	NN	O	O
an	NN	O	O
inactive	NN	O	O
HPRT	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
nonrandom	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
cytogenetically	NN	O	O
normal	NN	O	O
paternal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
and	NN	O	O
a	NN	O	O
microdeletion	NN	O	O
of	NN	O	O
the	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
on	NN	O	O
an	NN	O	O
active	NN	O	O
maternal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
were	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HPRT	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O
.	NN	O	O

Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
:	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
neurologic	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	I-Disease
myelin	NN	O	I-Disease
metabolism	NN	O	I-Disease
with	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
nosology	NN	O	O
of	NN	O	O
the	NN	O	O
inborn	NN	O	B-Disease
errors	NN	O	I-Disease
of	NN	O	I-Disease
myelin	NN	O	I-Disease
metabolism	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
stymied	NN	O	O
by	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
.	NN	O	O

Historically	NN	O	O
,	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
has	NN	O	O
encompassed	NN	O	O
a	NN	O	O
host	NN	O	O
of	NN	O	O
neurologic	NN	O	B-Disease
disorders	NN	O	I-Disease
that	NN	O	O
present	NN	O	O
with	NN	O	O
a	NN	O	O
deficit	NN	O	B-Disease
of	NN	O	I-Disease
myelin	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
membrane	NN	O	O
elaborated	NN	O	O
by	NN	O	O
glial	NN	O	O
cells	NN	O	O
that	NN	O	O
encircles	NN	O	O
and	NN	O	O
successively	NN	O	O
enwraps	NN	O	O
axons	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
a	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
pedigree	NN	O	O
of	NN	O	O
the	NN	O	O
classical	NN	O	O
type	NN	O	O
,	NN	O	O
with	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
inheritance	NN	O	O
,	NN	O	O
a	NN	O	O
typical	NN	O	O
clinical	NN	O	O
progression	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
pathologic	NN	O	O
loss	NN	O	O
of	NN	O	O
myelinating	NN	O	O
cells	NN	O	O
and	NN	O	O
myelin	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

To	NN	O	O
discriminate	NN	O	O
variants	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
oligonucleotide	NN	O	O
primers	NN	O	O
was	NN	O	O
constructed	NN	O	O
to	NN	O	O
polymerase	NN	O	O
-	NN	O	O
chain	NN	O	O
-	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
amplify	NN	O	O
and	NN	O	O
sequence	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
structural	NN	O	O
protein	NN	O	O
that	NN	O	O
comprises	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
of	NN	O	O
the	NN	O	O
myelin	NN	O	O
sheath	NN	O	O
.	NN	O	O

The	NN	O	O
PLP	NN	O	O
gene	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
the	NN	O	O
carrier	NN	O	O
mother	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
exhibited	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
more	NN	O	O
than	NN	O	O
2	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
sequenced	NN	O	O
,	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
that	NN	O	O
would	NN	O	O
create	NN	O	O
a	NN	O	O
serine	NN	O	O
substitution	NN	O	O
for	NN	O	O
proline	NN	O	O
at	NN	O	O
the	NN	O	O
carboxy	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
delineate	NN	O	O
the	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
define	NN	O	O
the	NN	O	O
possible	NN	O	O
molecular	NN	O	O
pathology	NN	O	O
of	NN	O	O
this	NN	O	O
dysmyelinating	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
address	NN	O	O
the	NN	O	O
molecular	NN	O	O
classification	NN	O	O
of	NN	O	O
inborn	NN	O	B-Disease
errors	NN	O	I-Disease
of	NN	O	I-Disease
myelin	NN	O	I-Disease
metabolism	NN	O	I-Disease
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
the	NN	O	O
classical	NN	O	O
form	NN	O	O
(	NN	O	O
type	NN	O	O
I	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
more	NN	O	O
severely	NN	O	O
affected	NN	O	O
,	NN	O	O
connatal	NN	O	O
variant	NN	O	O
of	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	I-Disease
type	NN	O	I-Disease
II	NN	O	I-Disease
)	NN	O	I-Disease
would	NN	O	O
be	NN	O	O
predicted	NN	O	O
to	NN	O	O
display	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
PLP	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
variants	NN	O	O
(	NN	O	O
types	NN	O	O
III	NN	O	O
-	NN	O	O
VI	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
have	NN	O	O
sometimes	NN	O	O
been	NN	O	O
categorized	NN	O	O
as	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
may	NN	O	O
represent	NN	O	O
mutations	NN	O	O
in	NN	O	O
genes	NN	O	O
encoding	NN	O	O
other	NN	O	O
structural	NN	O	O
myelin	NN	O	O
proteins	NN	O	O
or	NN	O	O
proteins	NN	O	O
critical	NN	O	O
to	NN	O	O
myelination	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
human	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
:	NN	O	O
analysis	NN	O	O
of	NN	O	O
platelet	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
mRNA	NN	O	O
.	NN	O	O

von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
vWD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
inherited	NN	O	B-Disease
bleeding	NN	O	I-Disease
disorder	NN	O	I-Disease
in	NN	O	O
humans	NN	O	O
,	NN	O	O
can	NN	O	O
result	NN	O	O
from	NN	O	O
either	NN	O	O
a	NN	O	O
quantitative	NN	O	O
or	NN	O	O
a	NN	O	O
qualitative	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
adhesive	NN	O	O
glycoprotein	NN	O	O
,	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
(	NN	O	O
vWF	NN	O	O
)	NN	O	O
.	NN	O	O

Molecular	NN	O	O
studies	NN	O	O
of	NN	O	O
vWD	NN	O	B-Disease
have	NN	O	O
been	NN	O	O
limited	NN	O	O
by	NN	O	O
the	NN	O	O
large	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
vWF	NN	O	O
gene	NN	O	O
and	NN	O	O
difficulty	NN	O	O
in	NN	O	O
obtaining	NN	O	O
the	NN	O	O
vWF	NN	O	O
mRNA	NN	O	O
from	NN	O	O
patients	NN	O	O
.	NN	O	O

By	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
adaptation	NN	O	O
of	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
,	NN	O	O
vWF	NN	O	O
mRNA	NN	O	O
was	NN	O	O
amplified	NN	O	O
and	NN	O	O
sequenced	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
platelets	NN	O	O
.	NN	O	O

A	NN	O	O
silent	NN	O	O
vWF	NN	O	O
allele	NN	O	O
was	NN	O	O
identified	NN	O	O
,	NN	O	O
resulting	NN	O	O
from	NN	O	O
a	NN	O	O
cis	NN	O	O
defect	NN	O	O
in	NN	O	O
vWF	NN	O	O
mRNA	NN	O	O
transcription	NN	O	O
or	NN	O	O
processing	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
type	NN	O	B-Disease
IIA	NN	O	I-Disease
vWD	NN	O	I-Disease
patients	NN	O	O
,	NN	O	O
two	NN	O	O
different	NN	O	O
but	NN	O	O
adjacent	NN	O	O
missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
the	NN	O	O
locations	NN	O	O
of	NN	O	O
which	NN	O	O
may	NN	O	O
identify	NN	O	O
an	NN	O	O
important	NN	O	O
vWF	NN	O	O
functional	NN	O	O
domain	NN	O	O
.	NN	O	O

Expression	NN	O	O
in	NN	O	O
heterologous	NN	O	O
cells	NN	O	O
of	NN	O	O
recombinant	NN	O	O
vWF	NN	O	O
containing	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
latter	NN	O	O
mutations	NN	O	O
reproduced	NN	O	O
the	NN	O	O
characteristic	NN	O	O
structural	NN	O	O
abnormality	NN	O	O
seen	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
IIA	NN	O	I-Disease
vWD	NN	O	I-Disease
plasma	NN	O	O
.	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
recurrent	NN	O	O
meningococcal	NN	O	B-Disease
infections	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
describe	NN	O	O
an	NN	O	O
11	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
girl	NN	O	O
suffering	NN	O	O
from	NN	O	O
recurrent	NN	O	B-Disease
meningitis	NN	O	I-Disease
with	NN	O	O
a	NN	O	O
complete	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C7	NN	O	O
)	NN	O	O
.	NN	O	O

Diagnosis	NN	O	O
was	NN	O	O
established	NN	O	O
by	NN	O	O
haemolytic	NN	O	O
titration	NN	O	O
and	NN	O	O
western	NN	O	O
blotting	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
serum	NN	O	O
lacked	NN	O	O
the	NN	O	O
85	NN	O	O
kDa	NN	O	O
C7	NN	O	O
chain	NN	O	O
.	NN	O	O

Haemolytic	NN	O	O
activity	NN	O	O
of	NN	O	O
serum	NN	O	O
was	NN	O	O
reconstituted	NN	O	O
with	NN	O	O
either	NN	O	O
pooled	NN	O	O
normal	NN	O	O
human	NN	O	O
serum	NN	O	O
or	NN	O	O
with	NN	O	O
purified	NN	O	O
C7	NN	O	O
.	NN	O	O

The	NN	O	O
relatives	NN	O	O
(	NN	O	O
parents	NN	O	O
and	NN	O	O
one	NN	O	O
sister	NN	O	O
)	NN	O	O
had	NN	O	O
half	NN	O	O
-	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
both	NN	O	O
immunochemically	NN	O	O
and	NN	O	O
functionally	NN	O	O
determined	NN	O	O
C7	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
heterozygous	NN	O	O
state	NN	O	O
for	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Translocation	NN	O	O
t	NN	O	O
(	NN	O	O
5	NN	O	O
;	NN	O	O
11	NN	O	O
)	NN	O	O
(	NN	O	O
q13	NN	O	O
.	NN	O	O
1	NN	O	O
;	NN	O	O
p13	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
familial	NN	O	O
isolated	NN	O	B-Disease
aniridia	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
father	NN	O	O
and	NN	O	O
daughter	NN	O	O
with	NN	O	O
isolated	NN	O	B-Disease
aniridia	NN	O	I-Disease
were	NN	O	O
observed	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
apparently	NN	O	O
balanced	NN	O	O
,	NN	O	O
reciprocal	NN	O	O
translocation	NN	O	O
involving	NN	O	O
chromosomes	NN	O	O
5	NN	O	O
and	NN	O	O
11	NN	O	O
[	NN	O	O
t	NN	O	O
(	NN	O	O
5	NN	O	O
;	NN	O	O
11	NN	O	O
)	NN	O	O
(	NN	O	O
q13	NN	O	O
.	NN	O	O
1	NN	O	O
;	NN	O	O
p13	NN	O	O
)	NN	O	O
]	NN	O	O
.	NN	O	O

No	NN	O	O
other	NN	O	O
clinical	NN	O	O
characteristics	NN	O	O
often	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
11p13	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
,	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
3	NN	O	O
other	NN	O	O
instances	NN	O	O
of	NN	O	O
single	NN	O	O
breaks	NN	O	O
at	NN	O	O
11p13	NN	O	O
and	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
supports	NN	O	O
the	NN	O	O
assignment	NN	O	O
of	NN	O	O
AN2	NN	O	O
to	NN	O	O
11p13	NN	O	O
.	NN	O	O

Homozygous	NN	O	B-Disease
hypobetalipoproteinemia	NN	O	I-Disease
:	NN	O	O
a	NN	O	O
disease	NN	O	O
distinct	NN	O	O
from	NN	O	O
abetalipoproproteinemia	NN	O	B-Disease
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
.	NN	O	O

apoB	NN	O	O
DNA	NN	O	O
,	NN	O	O
RNA	NN	O	O
,	NN	O	O
and	NN	O	O
protein	NN	O	O
from	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
homozygous	NN	O	B-Disease
hypobetalipoproteinemia	NN	O	I-Disease
(	NN	O	O
HBL	NN	O	B-Disease
)	NN	O	O
were	NN	O	O
evaluated	NN	O	O
and	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
individuals	NN	O	O
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
with	NN	O	O
10	NN	O	O
different	NN	O	O
cDNA	NN	O	O
probes	NN	O	O
revealed	NN	O	O
a	NN	O	O
normal	NN	O	O
gene	NN	O	O
without	NN	O	O
major	NN	O	O
insertions	NN	O	O
,	NN	O	O
deletions	NN	O	O
,	NN	O	O
or	NN	O	O
rearrangements	NN	O	O
.	NN	O	O

Northern	NN	O	O
and	NN	O	O
slot	NN	O	O
blot	NN	O	O
analyses	NN	O	O
of	NN	O	O
total	NN	O	O
liver	NN	O	O
mRNA	NN	O	O
from	NN	O	O
HBL	NN	O	B-Disease
patients	NN	O	O
documented	NN	O	O
a	NN	O	O
normal	NN	O	O
size	NN	O	O
apoB	NN	O	O
mRNA	NN	O	O
that	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
quantities	NN	O	O
.	NN	O	O

ApoB	NN	O	O
protein	NN	O	O
was	NN	O	O
detected	NN	O	O
within	NN	O	O
HBL	NN	O	B-Disease
hepatocytes	NN	O	O
utilizing	NN	O	O
immunohistochemical	NN	O	O
techniques	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
quantity	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
samples	NN	O	O
.	NN	O	O

No	NN	O	O
apoB	NN	O	O
was	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
of	NN	O	O
HBL	NN	O	B-Disease
individuals	NN	O	O
with	NN	O	O
an	NN	O	O
ELISA	NN	O	O
assay	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
most	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
apoB	NN	O	O
gene	NN	O	O
in	NN	O	O
HBL	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
an	NN	O	O
abnormal	NN	O	O
apoB	NN	O	O
protein	NN	O	O
and	NN	O	O
apoB	NN	O	O
mRNA	NN	O	O
instability	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
previously	NN	O	O
noted	NN	O	O
in	NN	O	O
abetalipoproteinemia	NN	O	B-Disease
,	NN	O	O
which	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
an	NN	O	O
elevated	NN	O	O
level	NN	O	O
of	NN	O	O
hepatic	NN	O	O
apoB	NN	O	O
mRNA	NN	O	O
and	NN	O	O
accumulation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
hepatic	NN	O	O
apoB	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Spontaneous	NN	O	O
reversion	NN	O	O
of	NN	O	O
novel	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
mutation	NN	O	O
by	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
an	NN	O	O
unusual	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
has	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
partial	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
duplication	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
the	NN	O	O
cloning	NN	O	O
and	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
HPRT	NN	O	O
cDNA	NN	O	O
which	NN	O	O
shows	NN	O	O
the	NN	O	O
precise	NN	O	O
duplication	NN	O	O
of	NN	O	O
exons	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
an	NN	O	O
internal	NN	O	O
duplication	NN	O	O
of	NN	O	O
16	NN	O	O
-	NN	O	O
20	NN	O	O
kilobases	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
gene	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
duplication	NN	O	O
was	NN	O	O
not	NN	O	O
generated	NN	O	O
by	NN	O	O
a	NN	O	O
single	NN	O	O
unequal	NN	O	O
crossing	NN	O	O
-	NN	O	O
over	NN	O	O
event	NN	O	O
between	NN	O	O
two	NN	O	O
normal	NN	O	O
HPRT	NN	O	O
alleles	NN	O	O
.	NN	O	O

Growth	NN	O	O
of	NN	O	O
Epstein	NN	O	B-Disease
-	NN	O	I-Disease
Barr	NN	O	I-Disease
virus	NN	O	O
-	NN	O	O
transformed	NN	O	O
lymphoblasts	NN	O	O
from	NN	O	O
this	NN	O	O
patient	NN	O	O
in	NN	O	O
selective	NN	O	O
medium	NN	O	O
has	NN	O	O
permitted	NN	O	O
isolation	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
HPRT	NN	O	O
+	NN	O	O
revertants	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
reversion	NN	O	O
event	NN	O	O
involves	NN	O	O
a	NN	O	O
second	NN	O	O
major	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
where	NN	O	O
most	NN	O	O
or	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
duplicated	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
gene	NN	O	O
is	NN	O	O
deleted	NN	O	O
.	NN	O	O

The	NN	O	O
original	NN	O	O
mutation	NN	O	O
therefore	NN	O	O
has	NN	O	O
the	NN	O	O
potential	NN	O	O
for	NN	O	O
spontaneous	NN	O	O
somatic	NN	O	O
reversion	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
relatively	NN	O	O
mild	NN	O	O
symptoms	NN	O	O
of	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
exhibited	NN	O	O
by	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O
.	NN	O	O

Complex	NN	O	B-Disease
glycerol	NN	O	I-Disease
kinase	NN	O	I-Disease
deficiency	NN	O	I-Disease
:	NN	O	O
molecular	NN	O	O
-	NN	O	O
genetic	NN	O	O
,	NN	O	O
cytogenetic	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
studies	NN	O	O
of	NN	O	O
five	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O

Five	NN	O	O
male	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
complex	NN	O	B-Disease
glycerol	NN	O	I-Disease
kinase	NN	O	I-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
CGKD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
their	NN	O	O
relatives	NN	O	O
were	NN	O	O
studied	NN	O	O
clinically	NN	O	O
,	NN	O	O
cytogenetically	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
-	NN	O	O
genetically	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
had	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
or	NN	O	O
muscle	NN	O	B-Disease
weakness	NN	O	I-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
congenital	NN	O	B-Disease
adrenal	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
glycerol	NN	O	B-Disease
kinase	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

High	NN	O	O
-	NN	O	O
resolution	NN	O	O
GTG	NN	O	O
-	NN	O	O
banded	NN	O	O
chromosomes	NN	O	O
showed	NN	O	O
a	NN	O	O
microdeletion	NN	O	O
in	NN	O	O
the	NN	O	O
Xp21	NN	O	O
region	NN	O	O
in	NN	O	O
all	NN	O	O
four	NN	O	O
patients	NN	O	O
examined	NN	O	O
and	NN	O	O
in	NN	O	O
all	NN	O	O
five	NN	O	O
mothers	NN	O	O
.	NN	O	O

Southern	NN	O	O
hybridizations	NN	O	O
,	NN	O	O
after	NN	O	O
digestions	NN	O	O
by	NN	O	O
restriction	NN	O	O
endonucleases	NN	O	O
,	NN	O	O
with	NN	O	O
various	NN	O	O
cloned	NN	O	O
DNAs	NN	O	O
(	NN	O	O
D2	NN	O	O
,	NN	O	O
99	NN	O	O
-	NN	O	O
6	NN	O	O
,	NN	O	O
B24	NN	O	O
,	NN	O	O
C7	NN	O	O
,	NN	O	O
L1	NN	O	O
-	NN	O	O
4	NN	O	O
,	NN	O	O
cDMD13	NN	O	O
-	NN	O	O
14	NN	O	O
,	NN	O	O
J66	NN	O	O
-	NN	O	O
HI	NN	O	O
,	NN	O	O
P20	NN	O	O
,	NN	O	O
J	NN	O	O
-	NN	O	O
Bir	NN	O	O
,	NN	O	O
ERT87	NN	O	O
-	NN	O	O
30	NN	O	O
,	NN	O	O
ERT87	NN	O	O
-	NN	O	O
15	NN	O	O
,	NN	O	O
ERT87	NN	O	O
-	NN	O	O
8	NN	O	O
,	NN	O	O
ERT87	NN	O	O
-	NN	O	O
1	NN	O	O
,	NN	O	O
XJ	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O
1	NN	O	O
,	NN	O	O
754	NN	O	O
,	NN	O	O
cx5	NN	O	O
.	NN	O	O
7	NN	O	O
,	NN	O	O
and	NN	O	O
OTC	NN	O	O
-	NN	O	O
1	NN	O	O
)	NN	O	O
that	NN	O	O
are	NN	O	O
located	NN	O	O
around	NN	O	O
Xp21	NN	O	O
also	NN	O	O
showed	NN	O	O
a	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
genome	NN	O	O
of	NN	O	O
all	NN	O	O
patients	NN	O	O
and	NN	O	O
mothers	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
deletion	NN	O	O
differed	NN	O	O
in	NN	O	O
size	NN	O	O
among	NN	O	O
patients	NN	O	O
,	NN	O	O
a	NN	O	O
segment	NN	O	O
commonly	NN	O	O
absent	NN	O	O
was	NN	O	O
located	NN	O	O
between	NN	O	O
the	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
corresponding	NN	O	O
to	NN	O	O
L1	NN	O	O
-	NN	O	O
4	NN	O	O
and	NN	O	O
cDMD13	NN	O	O
-	NN	O	O
14	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
glycerol	NN	O	O
kinase	NN	O	O
(	NN	O	O
GK	NN	O	O
)	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
this	NN	O	O
segment	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
five	NN	O	O
patients	NN	O	O
and	NN	O	O
reported	NN	O	O
CGKD	NN	O	B-Disease
or	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
patients	NN	O	O
with	NN	O	O
DNA	NN	O	O
deletion	NN	O	O
suggests	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
certain	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
gonadotropin	NN	O	B-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
GTD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
and	NN	O	O
results	NN	O	O
of	NN	O	O
previous	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
genes	NN	O	O
for	NN	O	O
ornithine	NN	O	O
transcarbamylase	NN	O	O
(	NN	O	O
OTC	NN	O	O
)	NN	O	O
,	NN	O	O
DMD	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
GK	NN	O	O
and	NN	O	O
putative	NN	O	O
genes	NN	O	O
responsible	NN	O	O
for	NN	O	O
congenital	NN	O	B-Disease
adrenal	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
(	NN	O	O
AHC	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
GTD	NN	O	B-Disease
are	NN	O	O
arranged	NN	O	O
from	NN	O	O
telomere	NN	O	O
to	NN	O	O
centromere	NN	O	O
as	NN	O	O
pter	NN	O	O
-	NN	O	O
-	NN	O	O
GTD	NN	O	O
-	NN	O	O
-	NN	O	O
AHC	NN	O	O
-	NN	O	O
-	NN	O	O
GK	NN	O	O
-	NN	O	O
-	NN	O	O
DMD	NN	O	O
-	NN	O	O
-	NN	O	O
OTC	NN	O	O
-	NN	O	O
-	NN	O	O
cen	NN	O	O

Genetically	NN	O	O
determined	NN	O	O
low	NN	O	O
C4	NN	O	O
:	NN	O	O
a	NN	O	O
predisposing	NN	O	O
factor	NN	O	O
to	NN	O	O
autoimmune	NN	O	B-Disease
chronic	NN	O	I-Disease
active	NN	O	I-Disease
hepatitis	NN	O	I-Disease
.	NN	O	O

Of	NN	O	O
26	NN	O	O
patients	NN	O	O
with	NN	O	O
autoimmune	NN	O	B-Disease
chronic	NN	O	I-Disease
active	NN	O	I-Disease
hepatitis	NN	O	I-Disease
(	NN	O	O
CAH	NN	O	B-Disease
)	NN	O	O
starting	NN	O	O
in	NN	O	O
childhood	NN	O	O
18	NN	O	O
(	NN	O	O
69	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
low	NN	O	O
C4	NN	O	O
and	NN	O	O
5	NN	O	O
(	NN	O	O
19	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
low	NN	O	O
C3	NN	O	O
serum	NN	O	O
levels	NN	O	O
.	NN	O	O

Impaired	NN	O	O
hepatic	NN	O	O
synthesis	NN	O	O
and	NN	O	O
immune	NN	O	O
-	NN	O	O
consumption	NN	O	O
were	NN	O	O
unlikely	NN	O	O
since	NN	O	O
transferrin	NN	O	O
levels	NN	O	O
were	NN	O	O
normal	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
,	NN	O	O
albumin	NN	O	O
levels	NN	O	O
were	NN	O	O
persistently	NN	O	O
low	NN	O	O
in	NN	O	O
only	NN	O	O
3	NN	O	O
,	NN	O	O
and	NN	O	O
only	NN	O	O
3	NN	O	O
had	NN	O	O
raised	NN	O	O
levels	NN	O	O
of	NN	O	O
activation	NN	O	O
fragment	NN	O	O
C3d	NN	O	O
.	NN	O	O

C4d	NN	O	O
was	NN	O	O
normal	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
studied	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
families	NN	O	O
of	NN	O	O
12	NN	O	O
probands	NN	O	O
with	NN	O	O
low	NN	O	O
C4	NN	O	O
,	NN	O	O
7	NN	O	O
parents	NN	O	O
had	NN	O	O
low	NN	O	O
C4	NN	O	O
and	NN	O	O
2	NN	O	O
had	NN	O	O
levels	NN	O	O
which	NN	O	O
were	NN	O	O
at	NN	O	O
the	NN	O	O
lower	NN	O	O
limit	NN	O	O
of	NN	O	O
normal	NN	O	O
.	NN	O	O

5	NN	O	O
of	NN	O	O
10	NN	O	O
siblings	NN	O	O
from	NN	O	O
5	NN	O	O
families	NN	O	O
had	NN	O	O
low	NN	O	O
C4	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
low	NN	O	O
C4	NN	O	O
levels	NN	O	O
in	NN	O	O
CAH	NN	O	B-Disease
are	NN	O	O
genetically	NN	O	O
determined	NN	O	O
.	NN	O	O

C4	NN	O	O
phenotyping	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
26	NN	O	O
parents	NN	O	O
showed	NN	O	O
that	NN	O	O
90	NN	O	O
%	NN	O	O
and	NN	O	O
81	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
had	NN	O	O
null	NN	O	O
allotypes	NN	O	O
at	NN	O	O
either	NN	O	O
the	NN	O	O
C4A	NN	O	O
or	NN	O	O
C4B	NN	O	O
locus	NN	O	O
compared	NN	O	O
with	NN	O	O
59	NN	O	O
%	NN	O	O
in	NN	O	O
controls	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
defective	NN	O	O
expression	NN	O	O
of	NN	O	O
structural	NN	O	O
genes	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
observed	NN	O	O
C4	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

An	NN	O	O
amino	NN	O	O
-	NN	O	O
acid	NN	O	O
substitution	NN	O	O
involved	NN	O	O
in	NN	O	O
phenylketonuria	NN	O	B-Disease
is	NN	O	O
in	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
with	NN	O	O
DNA	NN	O	O
haplotype	NN	O	O
2	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
human	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hepatic	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
,	NN	O	O
phenylalanine	NN	O	O
4	NN	O	O
-	NN	O	O
monooxygenase	NN	O	O
,	NN	O	O
EC	NN	O	O
1	NN	O	O
.	NN	O	O
14	NN	O	O
.	NN	O	O
16	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

PKU	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
common	NN	O	O
inborn	NN	O	B-Disease
error	NN	O	I-Disease
of	NN	O	I-Disease
amino	NN	O	I-Disease
-	NN	O	I-Disease
acid	NN	O	I-Disease
metabolism	NN	O	I-Disease
in	NN	O	O
caucasian	NN	O	O
populations	NN	O	O
and	NN	O	O
approximately	NN	O	O
1	NN	O	O
in	NN	O	O
50	NN	O	O
individuals	NN	O	O
are	NN	O	O
carriers	NN	O	O
of	NN	O	O
a	NN	O	O
PKU	NN	O	B-Disease
allele	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
PKU	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
twelve	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
-	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
RFLP	NN	O	O
)	NN	O	O
haplotypes	NN	O	O
of	NN	O	O
the	NN	O	O
PAH	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
northern	NN	O	O
European	NN	O	O
population	NN	O	O
and	NN	O	O
observed	NN	O	O
that	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
PKU	NN	O	B-Disease
alleles	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
are	NN	O	O
confined	NN	O	O
to	NN	O	O
four	NN	O	O
common	NN	O	O
RFLP	NN	O	O
haplotypes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
reported	NN	O	O
a	NN	O	O
splicing	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
RFLP	NN	O	O
haplotype	NN	O	O
3	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
at	NN	O	O
about	NN	O	O
40	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
the	NN	O	O
molecular	NN	O	B-Disease
lesion	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
the	NN	O	O
RFLP	NN	O	O
haplotype	NN	O	O
2	NN	O	O
mutant	NN	O	O
allele	NN	O	O
.	NN	O	O

This	NN	O	O
defect	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
exon	NN	O	O
12	NN	O	O
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
amino	NN	O	O
-	NN	O	O
acid	NN	O	O
substitution	NN	O	O
(	NN	O	O
Arg	NN	O	O
to	NN	O	O
Trp	NN	O	O
)	NN	O	O
at	NN	O	O
residue	NN	O	O
408	NN	O	O
of	NN	O	O
PAH	NN	O	O
.	NN	O	O

Direct	NN	O	O
hybridization	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
point	NN	O	O
mutation	NN	O	O
using	NN	O	O
a	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
mutation	NN	O	O
is	NN	O	O
also	NN	O	O
in	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
with	NN	O	O
RFLP	NN	O	O
haplotype	NN	O	O
2	NN	O	O
alleles	NN	O	O
that	NN	O	O
make	NN	O	O
up	NN	O	O
about	NN	O	O
20	NN	O	O
%	NN	O	O
of	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
genes	NN	O	O

Choroideremia	NN	O	B-Disease
:	NN	O	O
close	NN	O	O
linkage	NN	O	O
to	NN	O	O
DXYS1	NN	O	O
and	NN	O	O
DXYS12	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
segregation	NN	O	O
analysis	NN	O	O
and	NN	O	O
historical	NN	O	O
-	NN	O	O
genealogical	NN	O	O
evidence	NN	O	O
.	NN	O	O

Linkage	NN	O	O
studies	NN	O	O
using	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
were	NN	O	O
conducted	NN	O	O
in	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
choroideremia	NN	O	B-Disease
,	NN	O	O
designated	NN	O	O
TCD	NN	O	B-Disease
for	NN	O	O
Progressive	NN	O	B-Disease
Tapeto	NN	O	I-Disease
-	NN	O	I-Disease
Choroidal	NN	O	I-Disease
Dystrophy	NN	O	I-Disease
.	NN	O	O

Previously	NN	O	O
demonstrated	NN	O	O
close	NN	O	O
linkage	NN	O	O
with	NN	O	O
locus	NN	O	O
DXYS1	NN	O	O
was	NN	O	O
confirmed	NN	O	O
(	NN	O	O
lod	NN	O	O
11	NN	O	O
.	NN	O	O
44	NN	O	O
at	NN	O	O
0	NN	O	O
recombination	NN	O	O
distance	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
locus	NN	O	O
DXYS12	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
closely	NN	O	O
linked	NN	O	O
with	NN	O	O
TCD	NN	O	B-Disease
(	NN	O	O
lod	NN	O	O
3	NN	O	O
.	NN	O	O
31	NN	O	O
at	NN	O	O
0	NN	O	O
recombination	NN	O	O
distance	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
disease	NN	O	O
mainly	NN	O	O
occurs	NN	O	O
in	NN	O	O
three	NN	O	O
large	NN	O	O
kindreds	NN	O	O
in	NN	O	O
remote	NN	O	O
Northern	NN	O	O
Finland	NN	O	O
.	NN	O	O

While	NN	O	O
formal	NN	O	O
genealogical	NN	O	O
proof	NN	O	O
is	NN	O	O
lacking	NN	O	O
,	NN	O	O
all	NN	O	O
presently	NN	O	O
living	NN	O	O
(	NN	O	O
more	NN	O	O
than	NN	O	O
80	NN	O	O
affected	NN	O	O
males	NN	O	O
and	NN	O	O
120	NN	O	O
carrier	NN	O	O
females	NN	O	O
)	NN	O	O
probably	NN	O	O
originate	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
founder	NN	O	O
couple	NN	O	O
born	NN	O	O
in	NN	O	O
1644	NN	O	O
and	NN	O	O
1646	NN	O	O
,	NN	O	O
twelve	NN	O	O
generations	NN	O	O
ago	NN	O	O
.	NN	O	O

All	NN	O	O
36	NN	O	O
patients	NN	O	O
and	NN	O	O
48	NN	O	O
carriers	NN	O	O
tested	NN	O	O
from	NN	O	O
the	NN	O	O
three	NN	O	O
kindreds	NN	O	O
had	NN	O	O
the	NN	O	O
same	NN	O	O
haplotype	NN	O	O
(	NN	O	O
TCD	NN	O	B-Disease
/	NN	O	O
DXYS1	NN	O	O
,	NN	O	O
11kb	NN	O	O
/	NN	O	O
DXYS12	NN	O	O
,	NN	O	O
1	NN	O	O
.	NN	O	O
6kb	NN	O	O
)	NN	O	O
.	NN	O	O

Given	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
105	NN	O	O
female	NN	O	O
meioses	NN	O	O
transmitting	NN	O	O
TCD	NN	O	B-Disease
have	NN	O	O
occurred	NN	O	O
since	NN	O	O
1650	NN	O	O
in	NN	O	O
these	NN	O	O
kindreds	NN	O	O
,	NN	O	O
extremely	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
TCD	NN	O	B-Disease
,	NN	O	O
DXYS1	NN	O	O
and	NN	O	O
DXYS12	NN	O	O
is	NN	O	O
suggested	NN	O	O
.	NN	O	O

The	NN	O	O
above	NN	O	O
haplotype	NN	O	O
is	NN	O	O
a	NN	O	O
very	NN	O	O
useful	NN	O	O
diagnostic	NN	O	O
tool	NN	O	O
in	NN	O	O
these	NN	O	O
TCD	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
our	NN	O	O
historical	NN	O	O
-	NN	O	O
genealogical	NN	O	O
approach	NN	O	O
to	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
may	NN	O	O
be	NN	O	O
possible	NN	O	O
elsewhere	NN	O	O
in	NN	O	O
similar	NN	O	O
isolated	NN	O	O
populations	NN	O	O

Von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
disease	NN	O	I-Disease
maps	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
3	NN	O	O
associated	NN	O	O
with	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
.	NN	O	O

Von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
VHL	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
disorder	NN	O	I-Disease
with	NN	O	O
inherited	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
various	NN	O	O
forms	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
including	NN	O	O
hemangioblastomas	NN	O	B-Disease
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
phaeochromocytomas	NN	O	B-Disease
,	NN	O	O
pancreatic	NN	O	B-Disease
malignancies	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinomas	NN	O	I-Disease
.	NN	O	O

Renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinomas	NN	O	I-Disease
constitute	NN	O	O
a	NN	O	O
particularly	NN	O	O
frequent	NN	O	O
cause	NN	O	O
of	NN	O	O
death	NN	O	O
in	NN	O	O
this	NN	O	O
disorder	NN	O	O
,	NN	O	O
occurring	NN	O	O
as	NN	O	O
bilateral	NN	O	B-Disease
and	NN	O	I-Disease
multifocal	NN	O	I-Disease
tumours	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
presenting	NN	O	O
at	NN	O	O
an	NN	O	O
earlier	NN	O	O
age	NN	O	O
than	NN	O	O
in	NN	O	O
sporadic	NN	O	O
,	NN	O	O
non	NN	O	O
-	NN	O	O
familial	NN	O	O
cases	NN	O	O
of	NN	O	O
this	NN	O	O
tumour	NN	O	B-Disease
type	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
locus	NN	O	O
encoding	NN	O	O
the	NN	O	O
human	NN	O	O
homologoue	NN	O	O
of	NN	O	O
the	NN	O	O
RAF1	NN	O	O
oncogene	NN	O	O
,	NN	O	O
which	NN	O	O
maps	NN	O	O
to	NN	O	O
chromosome	NN	O	O
3p25	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
4	NN	O	O
)	NN	O	O
.	NN	O	O

Crossovers	NN	O	O
with	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
locus	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
defect	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
phenotype	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
RAF1	NN	O	O
gene	NN	O	O
itself	NN	O	O
.	NN	O	O

An	NN	O	O
alternative	NN	O	O
or	NN	O	O
prior	NN	O	O
event	NN	O	O
to	NN	O	O
oncogene	NN	O	O
activation	NN	O	O
in	NN	O	O
tumour	NN	O	B-Disease
formation	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
and	NN	O	I-Disease
sporadic	NN	O	I-Disease
forms	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Sporadic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinomas	NN	O	I-Disease
have	NN	O	O
previously	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
regions	NN	O	O
on	NN	O	O
chromosome	NN	O	O
3p	NN	O	O
(	NN	O	O
refs	NN	O	O
5	NN	O	O
,	NN	O	O
6	NN	O	O
)	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
sporadic	NN	O	O
and	NN	O	O
VHL	NN	O	O
-	NN	O	O
associated	NN	O	O
forms	NN	O	O
of	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
might	NN	O	O
both	NN	O	O
result	NN	O	O
from	NN	O	O
alterations	NN	O	O
causing	NN	O	O
loss	NN	O	O
of	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
on	NN	O	O
this	NN	O	O
chromosome	NN	O	O
.	NN	O	O
.	NN	O	O

Tightly	NN	O	O
linked	NN	O	O
flanking	NN	O	O
markers	NN	O	O
for	NN	O	O
the	NN	O	O
Lowe	NN	O	B-Disease
oculocerebrorenal	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
with	NN	O	O
application	NN	O	O
to	NN	O	O
carrier	NN	O	O
assessment	NN	O	O
.	NN	O	O

The	NN	O	O
Lowe	NN	O	B-Disease
oculocerebrorenal	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
OCRL	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
congenital	NN	O	O
cataract	NN	O	B-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
defective	NN	O	B-Disease
renal	NN	O	I-Disease
tubular	NN	O	I-Disease
function	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
map	NN	O	O
assignment	NN	O	O
of	NN	O	O
OCRL	NN	O	B-Disease
to	NN	O	O
Xq24	NN	O	O
-	NN	O	O
q26	NN	O	O
has	NN	O	O
been	NN	O	O
made	NN	O	O
previously	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
DXS42	NN	O	O
at	NN	O	O
Xq24	NN	O	O
-	NN	O	O
q26	NN	O	O
(	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
,	NN	O	O
z	NN	O	O
=	NN	O	O
5	NN	O	O
.	NN	O	O
09	NN	O	O
)	NN	O	O
and	NN	O	O
with	NN	O	O
DXS10	NN	O	O
at	NN	O	O
Xq26	NN	O	O
(	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
,	NN	O	O
z	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
45	NN	O	O
)	NN	O	O
.	NN	O	O

Two	NN	O	O
additional	NN	O	O
families	NN	O	O
were	NN	O	O
studied	NN	O	O
and	NN	O	O
three	NN	O	O
additional	NN	O	O
polymorphisms	NN	O	O
were	NN	O	O
identified	NN	O	O
at	NN	O	O
DXS42	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
35	NN	O	O
-	NN	O	O
kb	NN	O	O
sequence	NN	O	O
isolated	NN	O	O
with	NN	O	O
the	NN	O	O
probe	NN	O	O
detecting	NN	O	O
the	NN	O	O
original	NN	O	O
polymorphism	NN	O	O
at	NN	O	O
DXS42	NN	O	O
.	NN	O	O

With	NN	O	O
additional	NN	O	O
OCRL	NN	O	B-Disease
families	NN	O	O
made	NN	O	O
informative	NN	O	O
for	NN	O	O
DXS42	NN	O	O
,	NN	O	O
theta	NN	O	O
remained	NN	O	O
0	NN	O	O
with	NN	O	O
z	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O

63	NN	O	O
;	NN	O	O
and	NN	O	O
for	NN	O	O
DXS10	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

03	NN	O	O
and	NN	O	O
z	NN	O	O
=	NN	O	O
7	NN	O	O
.	NN	O	O

07	NN	O	O
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
placing	NN	O	O
OCRL	NN	O	O
at	NN	O	O
Xq25	NN	O	O
also	NN	O	O
comes	NN	O	O
from	NN	O	O
a	NN	O	O
female	NN	O	O
with	NN	O	O
Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
an	NN	O	O
X	NN	O	O
;	NN	O	O
3	NN	O	O
translocation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
Xq25	NN	O	O
breakpoint	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
OCRL	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
two	NN	O	O
linked	NN	O	O
markers	NN	O	O
.	NN	O	O

Each	NN	O	O
derivative	NN	O	O
chromosome	NN	O	O
was	NN	O	O
isolated	NN	O	O
away	NN	O	O
from	NN	O	O
its	NN	O	O
normal	NN	O	O
counterpart	NN	O	O
in	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
.	NN	O	O

DXS42	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
derivative	NN	O	O
chromosome	NN	O	O
X	NN	O	O
containing	NN	O	O
Xpterq25	NN	O	O
,	NN	O	O
and	NN	O	O
DXS10	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
derivative	NN	O	O
chromosome	NN	O	O
3	NN	O	O
containing	NN	O	O
Xq25	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O

The	NN	O	O
markers	NN	O	O
DXS10	NN	O	O
and	NN	O	O
DXS42	NN	O	O
therefore	NN	O	O
show	NN	O	O
tight	NN	O	O
linkage	NN	O	O
with	NN	O	O
OCRL	NN	O	B-Disease
in	NN	O	O
six	NN	O	O
families	NN	O	O
and	NN	O	O
flank	NN	O	O
the	NN	O	O
Xq25	NN	O	O
breakpoint	NN	O	O
in	NN	O	O
a	NN	O	O
female	NN	O	O
patient	NN	O	O
with	NN	O	O
an	NN	O	O
X	NN	O	O
;	NN	O	O
3	NN	O	O
translocation	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
flanking	NN	O	O
markers	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
assess	NN	O	O
OCRL	NN	O	B-Disease
carrier	NN	O	O
status	NN	O	O
in	NN	O	O
women	NN	O	O
at	NN	O	O
risk	NN	O	O
.	NN	O	O

Results	NN	O	O
,	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
carrier	NN	O	O
determination	NN	O	O
by	NN	O	O
ophthalmologic	NN	O	O
examination	NN	O	O
,	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
slit	NN	O	O
-	NN	O	O
lamp	NN	O	O
exam	NN	O	O
can	NN	O	O
be	NN	O	O
a	NN	O	O
sensitive	NN	O	O
and	NN	O	O
specific	NN	O	O
method	NN	O	O
of	NN	O	O
carrier	NN	O	O
determination	NN	O	O
in	NN	O	O
many	NN	O	O
cases	NN	O	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
hypoxanthine	NN	O	O
-	NN	O	O
guanine	NN	O	O
phosphoribosyltransferase	NN	O	O
gene	NN	O	O
(	NN	O	O
HPRTYale	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Complete	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hypoxanthine	NN	O	I-Disease
-	NN	O	I-Disease
guanine	NN	O	I-Disease
phosphoribosyltransferase	NN	O	I-Disease
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
causes	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Previous	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	O
form	NN	O	O
of	NN	O	O
HPRT	NN	O	O
,	NN	O	O
HPRTYale	NN	O	O
,	NN	O	O
from	NN	O	O
a	NN	O	O
subject	NN	O	O
with	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
revealed	NN	O	O
normal	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
concentrations	NN	O	O
,	NN	O	O
no	NN	O	O
residual	NN	O	O
catalytic	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
cathodal	NN	O	O
migration	NN	O	O
upon	NN	O	O
PAGE	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
HPRTYale	NN	O	O
cDNA	NN	O	O
.	NN	O	O

The	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
of	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
HPRTYale	NN	O	O
cDNA	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
substitution	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
HPRT	NN	O	O
cDNA	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
C	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
position	NN	O	O
211	NN	O	O
.	NN	O	O

This	NN	O	O
transversion	NN	O	O
predicts	NN	O	O
substitution	NN	O	O
of	NN	O	O
arginine	NN	O	O
for	NN	O	O
glycine	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
position	NN	O	O
71	NN	O	O
,	NN	O	O
explaining	NN	O	O
the	NN	O	O
cathodal	NN	O	O
migration	NN	O	O
of	NN	O	O
HPRTYale	NN	O	O
.	NN	O	O

Chou	NN	O	O
-	NN	O	O
Fasman	NN	O	O
secondary	NN	O	O
structure	NN	O	O
analysis	NN	O	O
predicts	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
beta	NN	O	O
-	NN	O	O
turn	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
containing	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

Inclusion	NN	O	O
of	NN	O	O
the	NN	O	O
bulky	NN	O	O
arginine	NN	O	O
side	NN	O	O
chain	NN	O	O
in	NN	O	O
place	NN	O	O
of	NN	O	O
glycine	NN	O	O
probably	NN	O	O
disrupts	NN	O	O
protein	NN	O	O
folding	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

Cloning	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
cDNA	NN	O	O
allows	NN	O	O
identification	NN	O	O
of	NN	O	O
specific	NN	O	O
mutations	NN	O	O
,	NN	O	O
provides	NN	O	O
insight	NN	O	O
into	NN	O	O
mutational	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
and	NN	O	O
facilitates	NN	O	O
structure	NN	O	O
-	NN	O	O
function	NN	O	O
analysis	NN	O	O
of	NN	O	O
mutant	NN	O	O
proteins	NN	O	O
.	NN	O	O
.	NN	O	O

Two	NN	O	O
point	NN	O	O
mutations	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
G6PD	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
Sardinia	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
gene	NN	O	O
encoding	NN	O	O
glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
is	NN	O	O
highly	NN	O	O
polymorphic	NN	O	O
;	NN	O	O
more	NN	O	O
than	NN	O	O
300	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
different	NN	O	O
geographical	NN	O	O
areas	NN	O	O
appears	NN	O	O
to	NN	O	O
have	NN	O	O
arisen	NN	O	O
through	NN	O	O
independent	NN	O	O
mutational	NN	O	O
events	NN	O	O
,	NN	O	O
but	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
population	NN	O	O
it	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
heterogeneous	NN	O	O
.	NN	O	O

One	NN	O	O
example	NN	O	O
is	NN	O	O
the	NN	O	O
island	NN	O	O
of	NN	O	O
Sardinia	NN	O	O
,	NN	O	O
where	NN	O	O
careful	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
four	NN	O	O
different	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
.	NN	O	O

We	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
four	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
from	NN	O	O
Sardinia	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
only	NN	O	O
two	NN	O	O
mutations	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
area	NN	O	O
one	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
Seattle	NN	O	I-Disease
-	NN	O	I-Disease
like	NN	O	I-Disease
phenotype	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
milder	NN	O	O
form	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
;	NN	O	O
the	NN	O	O
other	NN	O	O
mutation	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
all	NN	O	O
forms	NN	O	O
of	NN	O	O
very	NN	O	O
severe	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Sardinia	NN	O	O
and	NN	O	O
,	NN	O	O
possibly	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
.	NN	O	O
.	NN	O	O

Chronic	NN	O	B-Disease
nonspherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
(	NN	O	O
CNSHA	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
glucose	NN	O	B-Disease
6	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
familial	NN	O	B-Disease
amyloidotic	NN	O	I-Disease
polyneuropathy	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Molecular	NN	O	O
study	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
variant	NN	O	O
(	NN	O	O
G6PD	NN	O	O
Clinic	NN	O	O
)	NN	O	O
with	NN	O	O
markedly	NN	O	O
acidic	NN	O	O
pH	NN	O	O
optimum	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
variant	NN	O	O
with	NN	O	O
severe	NN	O	O
erythrocytic	NN	O	B-Disease
G6PD	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
unique	NN	O	O
pH	NN	O	O
optimum	NN	O	O
is	NN	O	O
described	NN	O	O
in	NN	O	O
a	NN	O	O
young	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	B-Disease
nonspherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
(	NN	O	O
CNSHA	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
familial	NN	O	B-Disease
amyloidotic	NN	O	I-Disease
polyneuropathy	NN	O	I-Disease
(	NN	O	O
FAP	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Chronic	NN	O	B-Disease
hemolysis	NN	O	I-Disease
was	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
infections	NN	O	O
,	NN	O	O
oxidant	NN	O	O
drugs	NN	O	O
or	NN	O	O
ingestion	NN	O	O
of	NN	O	O
faba	NN	O	O
beans	NN	O	O
.	NN	O	O

Residual	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
was	NN	O	O
about	NN	O	O
2	NN	O	O
.	NN	O	O

6	NN	O	O
%	NN	O	O
and	NN	O	O
63	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
activity	NN	O	O
in	NN	O	O
erythrocytes	NN	O	O
and	NN	O	O
leucocytes	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

A	NN	O	O
molecular	NN	O	O
study	NN	O	O
using	NN	O	O
standard	NN	O	O
methods	NN	O	O
showed	NN	O	O
G6PD	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
to	NN	O	O
have	NN	O	O
normal	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
(	NN	O	O
at	NN	O	O
pH	NN	O	O
7	NN	O	O
.	NN	O	O
0	NN	O	O
,	NN	O	O
8	NN	O	O
.	NN	O	O
0	NN	O	O
and	NN	O	O
8	NN	O	O
.	NN	O	O
8	NN	O	O
)	NN	O	O
,	NN	O	O
normal	NN	O	O
apparent	NN	O	O
affinity	NN	O	O
for	NN	O	O
substrates	NN	O	O
(	NN	O	O
Km	NN	O	O
,	NN	O	O
G6P	NN	O	O
and	NN	O	O
NADP	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
slightly	NN	O	O
abnormal	NN	O	O
utilization	NN	O	O
of	NN	O	O
substrate	NN	O	O
analogues	NN	O	O
(	NN	O	O
decreased	NN	O	O
deamino	NN	O	O
-	NN	O	O
NADP	NN	O	O
and	NN	O	O
increased	NN	O	O
2	NN	O	O
-	NN	O	O
deoxyglucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
utilization	NN	O	O
)	NN	O	O
.	NN	O	O

Heat	NN	O	O
stability	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
(	NN	O	O
8	NN	O	O
%	NN	O	O
of	NN	O	O
residual	NN	O	O
activity	NN	O	O
after	NN	O	O
20	NN	O	O
min	NN	O	O
of	NN	O	O
incubation	NN	O	O
at	NN	O	O
46	NN	O	O
degrees	NN	O	O
C	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
particular	NN	O	O
characteristic	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
was	NN	O	O
a	NN	O	O
biphasic	NN	O	O
pH	NN	O	O
curve	NN	O	O
with	NN	O	O
a	NN	O	O
greatly	NN	O	O
increased	NN	O	O
activity	NN	O	O
at	NN	O	O
low	NN	O	O
pH	NN	O	O
.	NN	O	O

Although	NN	O	O
molecular	NN	O	O
characteristics	NN	O	O
of	NN	O	O
this	NN	O	O
variant	NN	O	O
closely	NN	O	O
resemble	NN	O	O
those	NN	O	O
of	NN	O	O
G6PD	NN	O	O
Bangkok	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Duarte	NN	O	O
,	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
distinguished	NN	O	O
from	NN	O	O
these	NN	O	O
and	NN	O	O
all	NN	O	O
other	NN	O	O
previously	NN	O	O
reported	NN	O	O
variants	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
its	NN	O	O
unusual	NN	O	O
pH	NN	O	O
curve	NN	O	O
.	NN	O	O

Therefore	NN	O	O
the	NN	O	O
present	NN	O	O
variant	NN	O	O
has	NN	O	O
been	NN	O	O
designated	NN	O	O
G6PD	NN	O	O
Clinic	NN	O	O
to	NN	O	O
distinguish	NN	O	O
it	NN	O	O
from	NN	O	O
other	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
previously	NN	O	O
described	NN	O	O

Molecular	NN	O	O
detection	NN	O	O
of	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
that	NN	O	O
disrupt	NN	O	O
the	NN	O	O
putative	NN	O	O
retinoblastoma	NN	O	B-Disease
susceptibility	NN	O	O
locus	NN	O	O
.	NN	O	O

A	NN	O	O
candidate	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
with	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
properties	NN	O	O
predicted	NN	O	O
for	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-Disease
susceptibility	NN	O	O
(	NN	O	O
RB1	NN	O	O
)	NN	O	O
locus	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
(	NN	O	O
S	NN	O	O
.	NN	O	O
H	NN	O	O
.	NN	O	O
Friend	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O
Bernards	NN	O	O
,	NN	O	O
S	NN	O	O
.	NN	O	O
Rogelj	NN	O	O
,	NN	O	O
R	NN	O	O
.	NN	O	O
A	NN	O	O
.	NN	O	O
Weinberg	NN	O	O
,	NN	O	O
J	NN	O	O
.	NN	O	O
M	NN	O	O
.	NN	O	O
Rapaport	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O
M	NN	O	O
.	NN	O	O
Albert	NN	O	O
,	NN	O	O
and	NN	O	O
T	NN	O	O
.	NN	O	O
P	NN	O	O
.	NN	O	O
Dryja	NN	O	O
,	NN	O	O
Nature	NN	O	O
[	NN	O	O
London	NN	O	O
]	NN	O	O
323	NN	O	O
643	NN	O	O
-	NN	O	O
645	NN	O	O
,	NN	O	O
1986	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
large	NN	O	O
size	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
(	NN	O	O
ca	NN	O	O
.	NN	O	O
200	NN	O	O
kilobases	NN	O	O
[	NN	O	O
kb	NN	O	O
]	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
multiple	NN	O	O
dispersed	NN	O	O
exons	NN	O	O
(	NN	O	O
Wiggs	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
N	NN	O	O
.	NN	O	O
Engl	NN	O	O
.	NN	O	O
J	NN	O	O
.	NN	O	O
Med	NN	O	O
.	NN	O	O
318	NN	O	O
151	NN	O	O
-	NN	O	O
157	NN	O	O
,	NN	O	O
1988	NN	O	O
)	NN	O	O
complicate	NN	O	O
molecular	NN	O	O
screening	NN	O	O
strategies	NN	O	O
important	NN	O	O
in	NN	O	O
prenatal	NN	O	O
and	NN	O	O
presymptomatic	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
in	NN	O	O
carrier	NN	O	O
detection	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
used	NN	O	O
field	NN	O	O
inversion	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
FIGE	NN	O	O
)	NN	O	O
to	NN	O	O
construct	NN	O	O
a	NN	O	O
restriction	NN	O	O
map	NN	O	O
of	NN	O	O
approximately	NN	O	O
1	NN	O	O
,	NN	O	O
000	NN	O	O
kb	NN	O	O
of	NN	O	O
DNA	NN	O	O
surrounding	NN	O	O
the	NN	O	O
RB1	NN	O	O
locus	NN	O	O
and	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
three	NN	O	O
retinoblastoma	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

DNA	NN	O	O
probes	NN	O	O
from	NN	O	O
either	NN	O	O
the	NN	O	O
5	NN	O	O
or	NN	O	O
3	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
a	NN	O	O
250	NN	O	O
-	NN	O	O
kb	NN	O	O
EagI	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
in	NN	O	O
DNA	NN	O	O
from	NN	O	O
unaffected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Both	NN	O	O
probes	NN	O	O
identified	NN	O	O
an	NN	O	O
additional	NN	O	O
hybridizing	NN	O	O
fragment	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
from	NN	O	O
each	NN	O	O
patient	NN	O	O
,	NN	O	O
permitting	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
to	NN	O	O
be	NN	O	O
mapped	NN	O	O
within	NN	O	O
the	NN	O	O
cloned	NN	O	O
RB1	NN	O	O
gene	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
breakpoint	NN	O	O
in	NN	O	O
one	NN	O	O
translocation	NN	O	O
cell	NN	O	O
line	NN	O	O
allowed	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
to	NN	O	O
be	NN	O	O
oriented	NN	O	O
with	NN	O	O
its	NN	O	O
5	NN	O	O
end	NN	O	O
toward	NN	O	O
the	NN	O	O
centromere	NN	O	O
.	NN	O	O

The	NN	O	O
5	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
also	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
clustering	NN	O	O
of	NN	O	O
sites	NN	O	O
for	NN	O	O
several	NN	O	O
infrequently	NN	O	O
cleaving	NN	O	O
restriction	NN	O	O
enzymes	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
HpaII	NN	O	O
tiny	NN	O	O
fragment	NN	O	O
island	NN	O	O
.	NN	O	O

The	NN	O	O
detection	NN	O	O
and	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
all	NN	O	O
three	NN	O	O
retinoblastoma	NN	O	B-Disease
patients	NN	O	O
to	NN	O	O
within	NN	O	O
the	NN	O	O
putative	NN	O	O
RB1	NN	O	O
gene	NN	O	O
substantiated	NN	O	O
the	NN	O	O
authenticity	NN	O	O
of	NN	O	O
this	NN	O	O
candidate	NN	O	O
sequence	NN	O	O
and	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
FIGE	NN	O	O
in	NN	O	O
detecting	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
affecting	NN	O	O
this	NN	O	O
locus	NN	O	O
.	NN	O	O

Inherited	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
recurrent	NN	O	O
infections	NN	O	O
and	NN	O	O
glomerulonephritis	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
10	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
Laotian	NN	O	O
boy	NN	O	O
had	NN	O	O
homozygous	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
third	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
and	NN	O	O
recurrent	NN	O	O
bacterial	NN	O	B-Disease
infections	NN	O	I-Disease
beginning	NN	O	O
at	NN	O	O
age	NN	O	O
5	NN	O	O
months	NN	O	O
.	NN	O	O

Cellular	NN	O	O
and	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
were	NN	O	O
normal	NN	O	O
,	NN	O	O
as	NN	O	O
were	NN	O	O
polymorphonuclear	NN	O	O
leukocyte	NN	O	O
chemotaxis	NN	O	O
and	NN	O	O
bactericidal	NN	O	O
activities	NN	O	O
.	NN	O	O

Serum	NN	O	O
complement	NN	O	O
-	NN	O	O
mediated	NN	O	O
hemolytic	NN	O	O
,	NN	O	O
chemotactic	NN	O	O
,	NN	O	O
and	NN	O	O
opsonic	NN	O	O
activities	NN	O	O
were	NN	O	O
deficient	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
addition	NN	O	O
of	NN	O	O
purified	NN	O	O
C3	NN	O	O
to	NN	O	O
patient	NN	O	O
serum	NN	O	O
restored	NN	O	O
hemolytic	NN	O	O
complement	NN	O	O
to	NN	O	O
normal	NN	O	O
levels	NN	O	O
,	NN	O	O
and	NN	O	O
plasma	NN	O	O
infusion	NN	O	O
during	NN	O	O
each	NN	O	O
of	NN	O	O
four	NN	O	O
episodes	NN	O	O
of	NN	O	O
pneumonia	NN	O	B-Disease
significantly	NN	O	O
enhanced	NN	O	O
serum	NN	O	O
opsonic	NN	O	O
activity	NN	O	O
for	NN	O	O
as	NN	O	O
long	NN	O	O
as	NN	O	O
36	NN	O	O
hours	NN	O	O
.	NN	O	O

A	NN	O	O
renal	NN	O	O
biopsy	NN	O	O
specimen	NN	O	O
revealed	NN	O	O
mesangiopathic	NN	O	B-Disease
glomerulonephritis	NN	O	I-Disease
,	NN	O	O
although	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
circulating	NN	O	O
IgG	NN	O	O
immune	NN	O	O
complexes	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
further	NN	O	O
support	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
immune	NN	O	B-Disease
-	NN	O	I-Disease
complex	NN	O	I-Disease
disease	NN	O	I-Disease
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
plasma	NN	O	O
infusion	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
adjunct	NN	O	O
to	NN	O	O
antibiotic	NN	O	O
therapy	NN	O	O
in	NN	O	O
the	NN	O	O
management	NN	O	O
of	NN	O	O
severe	NN	O	O
pyogenic	NN	O	B-Disease
infections	NN	O	I-Disease
in	NN	O	O
patients	NN	O	O
with	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

DNA	NN	O	O
restriction	NN	O	O
fragments	NN	O	O
associated	NN	O	O
with	NN	O	O
alpha	NN	O	O
1	NN	O	O
-	NN	O	O
antitrypsin	NN	O	O
indicate	NN	O	O
a	NN	O	O
single	NN	O	O
origin	NN	O	O
for	NN	O	O
deficiency	NN	O	O
allele	NN	O	O
PI	NN	O	O
Z	NN	O	O
.	NN	O	O

The	NN	O	O
alpha	NN	O	O
1	NN	O	O
-	NN	O	O
protease	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
or	NN	O	O
alpha	NN	O	O
-	NN	O	O
antitrypsin	NN	O	O
(	NN	O	O
AAT	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	O
plasma	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
leukocyte	NN	O	O
elastase	NN	O	O
and	NN	O	O
bacterial	NN	O	O
proteases	NN	O	O
,	NN	O	O
is	NN	O	O
encoded	NN	O	O
at	NN	O	O
the	NN	O	O
PI	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
14	NN	O	O
(	NN	O	O
14q24	NN	O	O
.	NN	O	O
3	NN	O	O
-	NN	O	O
q32	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
AAT	NN	O	I-Disease
in	NN	O	O
individuals	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
PI	NN	O	O
Z	NN	O	O
allele	NN	O	O
occurs	NN	O	O
in	NN	O	O
about	NN	O	O
1	NN	O	O
in	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
-	NN	O	O
8	NN	O	O
,	NN	O	O
000	NN	O	O
caucasians	NN	O	O
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
early	NN	O	O
adult	NN	O	O
onset	NN	O	O
emphysema	NN	O	B-Disease
and	NN	O	O
liver	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
used	NN	O	O
DNA	NN	O	O
polymorphisms	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
AAT	NN	O	O
gene	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
PI	NN	O	O
Z	NN	O	O
allele	NN	O	O
.	NN	O	O

Using	NN	O	O
two	NN	O	O
genomic	NN	O	O
probes	NN	O	O
extending	NN	O	O
into	NN	O	O
the	NN	O	O
5	NN	O	O
and	NN	O	O
3	NN	O	O
flanking	NN	O	O
regions	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
eight	NN	O	O
polymorphic	NN	O	O
restriction	NN	O	O
sites	NN	O	O
.	NN	O	O

Extensive	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
occurs	NN	O	O
throughout	NN	O	O
the	NN	O	O
probed	NN	O	O
region	NN	O	O
with	NN	O	O
the	NN	O	O
PI	NN	O	O
Z	NN	O	O
allele	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
normal	NN	O	O
PI	NN	O	O
M	NN	O	O
alleles	NN	O	O
.	NN	O	O

The	NN	O	O
Z	NN	O	O
allele	NN	O	O
occurs	NN	O	O
mainly	NN	O	O
with	NN	O	O
one	NN	O	O
haplotype	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
single	NN	O	O
,	NN	O	O
relatively	NN	O	O
recent	NN	O	O
,	NN	O	O
origin	NN	O	O
in	NN	O	O
caucasians	NN	O	O

Segregation	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
marker	NN	O	O
localised	NN	O	O
Xp21	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
Xp21	NN	O	O
.	NN	O	O
3	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

A	NN	O	O
DNA	NN	O	O
marker	NN	O	O
C7	NN	O	O
,	NN	O	O
localised	NN	O	O
Xp21	NN	O	O
.	NN	O	O

1	NN	O	O
-	NN	O	O
Xp21	NN	O	O
.	NN	O	O

3	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
studied	NN	O	O
in	NN	O	O
kindreds	NN	O	O
segregating	NN	O	O
for	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
DMD	NN	O	B-Disease
families	NN	O	O
four	NN	O	O
crossovers	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
38	NN	O	O
informative	NN	O	O
meioses	NN	O	O
between	NN	O	O
C7	NN	O	O
and	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
(	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
12	NN	O	O
,	NN	O	O
z	NN	O	O
max	NN	O	O
=	NN	O	O
+	NN	O	O
2	NN	O	O
.	NN	O	O
72	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
BMD	NN	O	B-Disease
families	NN	O	O
no	NN	O	O
recombinants	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
16	NN	O	O
informative	NN	O	O
meioses	NN	O	O
studied	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
localisation	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
in	NN	O	O
these	NN	O	O
disorders	NN	O	O
being	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
region	NN	O	O
of	NN	O	O
Xp21	NN	O	O
.	NN	O	O

Studies	NN	O	O
in	NN	O	O
families	NN	O	O
also	NN	O	O
segregating	NN	O	O
for	NN	O	O
the	NN	O	O
DNA	NN	O	O
marker	NN	O	O
754	NN	O	O
support	NN	O	O
the	NN	O	O
previously	NN	O	O
reported	NN	O	O
physical	NN	O	O
order	NN	O	O
of	NN	O	O
these	NN	O	O
loci	NN	O	O
as	NN	O	O
X	NN	O	O
centromere	NN	O	O
-	NN	O	O
754	NN	O	O
-	NN	O	O
DMD	NN	O	O
-	NN	O	O
BMD	NN	O	O
-	NN	O	O
C7	NN	O	O
-	NN	O	O
X	NN	O	O
telomere	NN	O	O
.	NN	O	O

A	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

11	NN	O	O
(	NN	O	O
z	NN	O	O
max	NN	O	O
=	NN	O	O
+	NN	O	O
5	NN	O	O
.	NN	O	O
58	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
DMD	NN	O	O
-	NN	O	O
754	NN	O	O
by	NN	O	O
combining	NN	O	O
our	NN	O	O
previously	NN	O	O
published	NN	O	O
data	NN	O	O
with	NN	O	O
the	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
.	NN	O	O

C7	NN	O	O
and	NN	O	O
754	NN	O	O
thus	NN	O	O
provide	NN	O	O
good	NN	O	O
bridging	NN	O	O
markers	NN	O	O
for	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
BMD	NN	O	B-Disease

Isolation	NN	O	O
of	NN	O	O
molecular	NN	O	O
probes	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
chromosome	NN	O	O
15	NN	O	O
instability	NN	O	O
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
recombinant	NN	O	O
DNA	NN	O	O
techniques	NN	O	O
have	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
obtain	NN	O	O
reagents	NN	O	O
for	NN	O	O
a	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

HindIII	NN	O	O
total	NN	O	O
-	NN	O	O
digest	NN	O	O
libraries	NN	O	O
were	NN	O	O
prepared	NN	O	O
in	NN	O	O
lambda	NN	O	O
phage	NN	O	O
Charon	NN	O	O
21A	NN	O	O
from	NN	O	O
flow	NN	O	O
-	NN	O	O
sorted	NN	O	O
inverted	NN	O	O
duplicated	NN	O	O
no	NN	O	O
.	NN	O	O
15	NN	O	O
human	NN	O	O
chromosomes	NN	O	O
and	NN	O	O
propagated	NN	O	O
on	NN	O	O
recombination	NN	O	O
-	NN	O	O
proficient	NN	O	O
(	NN	O	O
LE392	NN	O	O
)	NN	O	O
and	NN	O	O
recBC	NN	O	O
-	NN	O	O
,	NN	O	O
sbcB	NN	O	O
-	NN	O	O
(	NN	O	O
DB1257	NN	O	O
)	NN	O	O
bacteria	NN	O	O
.	NN	O	O

Twelve	NN	O	O
distinct	NN	O	O
chromosome	NN	O	O
15	NN	O	O
-	NN	O	O
specific	NN	O	O
probes	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
.	NN	O	O

Eight	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
15q11	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
13	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
these	NN	O	O
eight	NN	O	O
sublocalized	NN	O	O
to	NN	O	O
band	NN	O	O
15q11	NN	O	O
.	NN	O	O

2	NN	O	O
and	NN	O	O
are	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
deleted	NN	O	O
in	NN	O	O
DNA	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
patients	NN	O	O
examined	NN	O	O
with	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Heteroduplex	NN	O	O
analysis	NN	O	O
of	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
clones	NN	O	O
,	NN	O	O
which	NN	O	O
grew	NN	O	O
on	NN	O	O
DB1257	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
LE392	NN	O	O
,	NN	O	O
revealed	NN	O	O
stem	NN	O	O
-	NN	O	O
loop	NN	O	O
structures	NN	O	O
in	NN	O	O
the	NN	O	O
inserts	NN	O	O
,	NN	O	O
indicative	NN	O	O
of	NN	O	O
inverted	NN	O	O
,	NN	O	O
repeated	NN	O	O
DNA	NN	O	O
elements	NN	O	O
.	NN	O	O

Such	NN	O	O
DNA	NN	O	O
repeats	NN	O	O
might	NN	O	O
account	NN	O	O
for	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
cloning	NN	O	O
instability	NN	O	O
of	NN	O	O
DNA	NN	O	O
segments	NN	O	O
from	NN	O	O
proximal	NN	O	O
15q	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
and	NN	O	O
physical	NN	O	O
instability	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
repeated	NN	O	O
sequences	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
from	NN	O	O
band	NN	O	O
15q11	NN	O	O
.	NN	O	O

2	NN	O	O
may	NN	O	O
elucidate	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
instability	NN	O	O
of	NN	O	O
this	NN	O	O
chromosomal	NN	O	O
region	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
or	NN	O	O
other	NN	O	O
diseases	NN	O	O
associated	NN	O	O
with	NN	O	O
chromosomal	NN	O	B-Disease
abnormalities	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
proximal	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
15	NN	O	O

Analysis	NN	O	O
of	NN	O	O
deletions	NN	O	O
in	NN	O	O
DNA	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
Becker	NN	O	B-Disease
and	NN	O	I-Disease
Duchenne	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
for	NN	O	O
which	NN	O	O
the	NN	O	O
biochemical	NN	O	O
defect	NN	O	O
is	NN	O	O
as	NN	O	O
yet	NN	O	O
unknown	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
two	NN	O	O
cloned	NN	O	O
segments	NN	O	O
of	NN	O	O
human	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
DNA	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
which	NN	O	O
detect	NN	O	O
structural	NN	O	O
alterations	NN	O	O
within	NN	O	O
or	NN	O	O
near	NN	O	O
the	NN	O	O
genetic	NN	O	O
locus	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
disorder	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
cloned	NN	O	O
segments	NN	O	O
were	NN	O	O
described	NN	O	O
as	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
locus	NN	O	O
and	NN	O	O
were	NN	O	O
capable	NN	O	O
of	NN	O	O
detecting	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
boys	NN	O	O
affected	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
determine	NN	O	O
more	NN	O	O
precisely	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
these	NN	O	O
deletions	NN	O	O
within	NN	O	O
a	NN	O	O
large	NN	O	O
population	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
the	NN	O	O
accuracy	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
segments	NN	O	O
,	NN	O	O
DXS164	NN	O	O
(	NN	O	O
pERT87	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
determining	NN	O	O
the	NN	O	O
inheritance	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
the	NN	O	O
subclones	NN	O	O
1	NN	O	O
,	NN	O	O
8	NN	O	O
and	NN	O	O
15	NN	O	O
were	NN	O	O
made	NN	O	O
available	NN	O	O
to	NN	O	O
many	NN	O	O
investigators	NN	O	O
throughout	NN	O	O
the	NN	O	O
world	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
combined	NN	O	O
results	NN	O	O
of	NN	O	O
more	NN	O	O
than	NN	O	O
20	NN	O	O
research	NN	O	O
laboratories	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
deletions	NN	O	O
at	NN	O	O
the	NN	O	O
DXS164	NN	O	O
locus	NN	O	O
in	NN	O	O
DNA	NN	O	O
samples	NN	O	O
isolated	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
DXS164	NN	O	O
locus	NN	O	O
apparently	NN	O	O
recombines	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
5	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
time	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
probably	NN	O	O
located	NN	O	O
between	NN	O	O
independent	NN	O	O
sites	NN	O	O
of	NN	O	O
mutation	NN	O	O
which	NN	O	O
yield	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
some	NN	O	O
deletions	NN	O	O
are	NN	O	O
delineated	NN	O	O
within	NN	O	O
the	NN	O	O
DXS164	NN	O	O
locus	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
evident	NN	O	O
that	NN	O	O
the	NN	O	O
deletions	NN	O	O
at	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
are	NN	O	O
frequent	NN	O	O
and	NN	O	O
extremely	NN	O	O
large	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
potential	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
through	NN	O	O
introduction	NN	O	O
of	NN	O	O
HPRT	NN	O	O
mutations	NN	O	O
into	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
neurological	NN	O	B-Disease
and	NN	O	I-Disease
behavioural	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
affecting	NN	O	O
only	NN	O	O
males	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
purine	NN	O	O
salvage	NN	O	O
enzyme	NN	O	O
hypoxanthine	NN	O	O
-	NN	O	O
guanosine	NN	O	O
phosphoribosyl	NN	O	O
transferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
.	NN	O	O

How	NN	O	O
the	NN	O	O
resulting	NN	O	O
alterations	NN	O	O
in	NN	O	O
purine	NN	O	O
metabolism	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
severe	NN	O	O
symptoms	NN	O	O
characteristic	NN	O	O
of	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patients	NN	O	O
is	NN	O	O
still	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

No	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
Hprt	NN	O	O
locus	NN	O	O
leading	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
activity	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
laboratory	NN	O	O
animals	NN	O	O
.	NN	O	O

To	NN	O	O
derive	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
cultured	NN	O	O
mouse	NN	O	O
embryonic	NN	O	O
stem	NN	O	O
cells	NN	O	O
,	NN	O	O
mutagenized	NN	O	O
by	NN	O	O
retroviral	NN	O	O
insertion	NN	O	O
and	NN	O	O
selected	NN	O	O
for	NN	O	O
loss	NN	O	O
of	NN	O	O
HPRT	NN	O	O
activity	NN	O	O
,	NN	O	O
to	NN	O	O
construct	NN	O	O
chimaeric	NN	O	O
mice	NN	O	O
.	NN	O	O

Two	NN	O	O
clonal	NN	O	O
lines	NN	O	O
carrying	NN	O	O
different	NN	O	O
mutant	NN	O	O
Hprt	NN	O	O
alleles	NN	O	O
have	NN	O	O
given	NN	O	O
rise	NN	O	O
to	NN	O	O
germ	NN	O	O
cells	NN	O	O
in	NN	O	O
chimaeras	NN	O	O
,	NN	O	O
allowing	NN	O	O
the	NN	O	O
derivation	NN	O	O
of	NN	O	O
strains	NN	O	O
of	NN	O	O
mutant	NN	O	O
mice	NN	O	O
having	NN	O	O
the	NN	O	O
same	NN	O	O
biochemical	NN	O	O
defect	NN	O	O
as	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Male	NN	O	O
mice	NN	O	O
carrying	NN	O	O
the	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
are	NN	O	O
viable	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
their	NN	O	O
cells	NN	O	O
shows	NN	O	O
a	NN	O	O
total	NN	O	O
lack	NN	O	O
of	NN	O	O
HPRT	NN	O	O
activity	NN	O	O
.	NN	O	O
.	NN	O	O

Re	NN	O	O
-	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
sublocalization	NN	O	O
of	NN	O	O
esterase	NN	O	O
D	NN	O	O
and	NN	O	O
its	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-Disease
locus	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
probe	NN	O	O
for	NN	O	O
the	NN	O	O
esterase	NN	O	O
D	NN	O	O
gene	NN	O	O
(	NN	O	O
ESD	NN	O	O
)	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
on	NN	O	O
human	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

The	NN	O	O
probe	NN	O	O
hybridized	NN	O	O
most	NN	O	O
strongly	NN	O	O
to	NN	O	O
13q14	NN	O	O
.	NN	O	O

2	NN	O	O
and	NN	O	O
13q14	NN	O	O
.	NN	O	O

3	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
raises	NN	O	O
doubts	NN	O	O
concerning	NN	O	O
the	NN	O	O
most	NN	O	O
recently	NN	O	O
published	NN	O	O
assignment	NN	O	O
of	NN	O	O
ESD	NN	O	O
to	NN	O	O
13q14	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

A	NN	O	O
deletion	NN	O	O
in	NN	O	O
an	NN	O	O
individual	NN	O	O
with	NN	O	O
retinoblastoma	NN	O	B-Disease
was	NN	O	O
reported	NN	O	O
to	NN	O	O
separate	NN	O	O
the	NN	O	O
closely	NN	O	O
linked	NN	O	O
ESD	NN	O	O
and	NN	O	O
retinoblastoma	NN	O	B-Disease
(	NN	O	O
RB1	NN	O	O
)	NN	O	O
loci	NN	O	O
,	NN	O	O
placing	NN	O	O
ESD	NN	O	O
proximal	NN	O	O
to	NN	O	O
RB1	NN	O	O
.	NN	O	O

Quantitative	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
studies	NN	O	O
of	NN	O	O
this	NN	O	O
deletion	NN	O	O
do	NN	O	O
not	NN	O	O
confirm	NN	O	O
this	NN	O	O
interpretation	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
they	NN	O	O
suggest	NN	O	O
that	NN	O	O
ESD	NN	O	O
is	NN	O	O
missing	NN	O	O
from	NN	O	O
the	NN	O	O
deleted	NN	O	O
chromosome	NN	O	O
13	NN	O	O
and	NN	O	O
duplicated	NN	O	O
on	NN	O	O
the	NN	O	O
normal	NN	O	O
homolog	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
deletion	NN	O	O
in	NN	O	O
this	NN	O	O
individual	NN	O	O
cannot	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
orientation	NN	O	O
nor	NN	O	O
the	NN	O	O
sublocalization	NN	O	O
of	NN	O	O
ESD	NN	O	O
and	NN	O	O
RB1	NN	O	O
within	NN	O	O
the	NN	O	O
13q14	NN	O	O
region	NN	O	O
.	NN	O	O

Hereditary	NN	O	B-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
common	NN	O	O
variable	NN	O	O
immunodeficiency	NN	O	B-Disease
.	NN	O	O

Homozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
19	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
boy	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
variable	NN	O	O
immunodeficiency	NN	O	B-Disease
manifested	NN	O	O
by	NN	O	O
marked	NN	O	O
hypoimmunoglobulinemia	NN	O	B-Disease
and	NN	O	O
impaired	NN	O	O
antibody	NN	O	O
responses	NN	O	O
,	NN	O	O
normal	NN	O	O
circulating	NN	O	O
B	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
and	NN	O	O
subnormal	NN	O	O
T	NN	O	O
-	NN	O	O
cell	NN	O	O
functions	NN	O	O
.	NN	O	O

Neither	NN	O	O
antilymphocytic	NN	O	O
autoantibodies	NN	O	O
nor	NN	O	O
chromosomal	NN	O	B-Disease
abnormalities	NN	O	I-Disease
were	NN	O	O
found	NN	O	O
.	NN	O	O

Serum	NN	O	O
immunoglobulin	NN	O	O
levels	NN	O	O
were	NN	O	O
within	NN	O	O
normal	NN	O	O
limits	NN	O	O
in	NN	O	O
his	NN	O	O
parents	NN	O	O
and	NN	O	O
brother	NN	O	O
who	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
patients	NN	O	O
lymphocytes	NN	O	O
were	NN	O	O
homozygous	NN	O	O
at	NN	O	O
the	NN	O	O
HLA	NN	O	O
-	NN	O	O
D	NN	O	O
locus	NN	O	O
but	NN	O	O
expressed	NN	O	O
an	NN	O	O
antigen	NN	O	O
different	NN	O	O
from	NN	O	O
DW2	NN	O	O
.	NN	O	O
.	NN	O	O

Mild	NN	O	O
and	NN	O	O
severe	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
deletions	NN	O	O
in	NN	O	O
Xp21	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analysed	NN	O	O
over	NN	O	O
300	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
Duchenne	NN	O	B-Disease
or	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
or	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Deletions	NN	O	O
have	NN	O	O
been	NN	O	O
characterised	NN	O	O
which	NN	O	O
encompass	NN	O	O
either	NN	O	O
the	NN	O	O
pERT87	NN	O	O
(	NN	O	O
DXS164	NN	O	O
)	NN	O	O
locus	NN	O	O
only	NN	O	O
,	NN	O	O
the	NN	O	O
XJ1	NN	O	O
.	NN	O	O

1	NN	O	O
(	NN	O	O
DXS206	NN	O	O
)	NN	O	O
and	NN	O	O
HIP25	NN	O	O
loci	NN	O	O
only	NN	O	O
,	NN	O	O
or	NN	O	O
all	NN	O	O
three	NN	O	O
loci	NN	O	O
.	NN	O	O

These	NN	O	O
loci	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
lie	NN	O	O
within	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
region	NN	O	O
covering	NN	O	O
several	NN	O	O
hundred	NN	O	O
kilobases	NN	O	O
(	NN	O	O
kb	NN	O	O
)	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

One	NN	O	O
mildly	NN	O	O
affected	NN	O	O
BMD	NN	O	B-Disease
patient	NN	O	O
possesses	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
110	NN	O	O
kb	NN	O	O
including	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Other	NN	O	O
patients	NN	O	O
with	NN	O	O
similar	NN	O	O
exon	NN	O	O
deletions	NN	O	O
,	NN	O	O
or	NN	O	O
smaller	NN	O	O
deletions	NN	O	O
,	NN	O	O
show	NN	O	O
the	NN	O	O
more	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
typical	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
these	NN	O	O
studies	NN	O	O
that	NN	O	O
the	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
cannot	NN	O	O
be	NN	O	O
explained	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
.	NN	O	O

We	NN	O	O
discuss	NN	O	O
this	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
candidate	NN	O	O
gene	NN	O	O
sequences	NN	O	O
.	NN	O	O

Patterns	NN	O	O
of	NN	O	O
exon	NN	O	O
deletions	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
panel	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
screened	NN	O	O
with	NN	O	O
the	NN	O	O
cDNA	NN	O	O
probes	NN	O	O
Cf56a	NN	O	O
and	NN	O	O
Cf23a	NN	O	O
,	NN	O	O
which	NN	O	O
detect	NN	O	O
exons	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

One	NN	O	O
or	NN	O	O
more	NN	O	O
exons	NN	O	O
were	NN	O	O
deleted	NN	O	O
in	NN	O	O
60	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
deletions	NN	O	O
were	NN	O	O
mapped	NN	O	O
and	NN	O	O
prove	NN	O	O
to	NN	O	O
be	NN	O	O
heterogeneous	NN	O	O
in	NN	O	O
size	NN	O	O
and	NN	O	O
extent	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

Deletions	NN	O	O
specific	NN	O	O
to	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
to	NN	O	O
BMD	NN	O	B-Disease
are	NN	O	O
described	NN	O	O
.	NN	O	O

Half	NN	O	O
of	NN	O	O
all	NN	O	O
BMD	NN	O	B-Disease
patients	NN	O	O
have	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
one	NN	O	O
particular	NN	O	O
small	NN	O	O
group	NN	O	O
of	NN	O	O
exons	NN	O	O
;	NN	O	O
smaller	NN	O	O
deletions	NN	O	O
within	NN	O	O
this	NN	O	O
same	NN	O	O
group	NN	O	O
produce	NN	O	O
the	NN	O	O
more	NN	O	O
severe	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Hereditary	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
third	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
fever	NN	O	B-Disease
,	NN	O	O
skin	NN	O	B-Disease
rash	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
arthralgias	NN	O	B-Disease
:	NN	O	O
response	NN	O	O
to	NN	O	O
transfusion	NN	O	O
of	NN	O	O
whole	NN	O	O
blood	NN	O	O
.	NN	O	O

A	NN	O	O
previously	NN	O	O
well	NN	O	O
34	NN	O	O
-	NN	O	O
month	NN	O	O
-	NN	O	O
old	NN	O	O
male	NN	O	O
presenting	NN	O	O
with	NN	O	O
fever	NN	O	B-Disease
,	NN	O	O
skin	NN	O	B-Disease
rash	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
arthralgias	NN	O	B-Disease
was	NN	O	O
found	NN	O	O
to	NN	O	O
lack	NN	O	O
C3	NN	O	O
by	NN	O	O
immunochemical	NN	O	O
(	NN	O	O
undetectable	NN	O	O
)	NN	O	O
and	NN	O	O
hemolytic	NN	O	O
(	NN	O	O
1	NN	O	O
%	NN	O	O
normal	NN	O	O
)	NN	O	O
assays	NN	O	O
.	NN	O	O

No	NN	O	O
infectious	NN	O	O
agent	NN	O	O
could	NN	O	O
be	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

Protein	NN	O	O
levels	NN	O	O
of	NN	O	O
Clq	NN	O	O
.	NN	O	O

C4	NN	O	O
,	NN	O	O
C5	NN	O	O
,	NN	O	O
properdin	NN	O	O
,	NN	O	O
and	NN	O	O
C3b	NN	O	O
-	NN	O	O
INA	NN	O	O
and	NN	O	O
hemolytic	NN	O	O
activities	NN	O	O
of	NN	O	O
complement	NN	O	O
components	NN	O	O
C1	NN	O	O
to	NN	O	O
C9	NN	O	O
except	NN	O	O
C3	NN	O	O
were	NN	O	O
normal	NN	O	O
or	NN	O	O
elevated	NN	O	O
;	NN	O	O
total	NN	O	O
hemolytic	NN	O	O
complement	NN	O	O
activity	NN	O	O
was	NN	O	O
13	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
was	NN	O	O
reconstituted	NN	O	O
by	NN	O	O
purified	NN	O	O
C3	NN	O	O
.	NN	O	O

Properdin	NN	O	O
factor	NN	O	O
B	NN	O	O
was	NN	O	O
702	NN	O	O
(	NN	O	O
normal	NN	O	O
175	NN	O	O
to	NN	O	O
275	NN	O	O
)	NN	O	O
mug	NN	O	O
/	NN	O	O
ml	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
cleaver	NN	O	O
upon	NN	O	O
addition	NN	O	O
of	NN	O	O
zymosan	NN	O	O
or	NN	O	O
cobra	NN	O	O
venom	NN	O	O
factor	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	O
had	NN	O	O
normal	NN	O	O
immune	NN	O	O
adherence	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
deficient	NN	O	O
in	NN	O	O
ability	NN	O	O
to	NN	O	O
opsonize	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
for	NN	O	O
uptake	NN	O	O
and	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
for	NN	O	O
killing	NN	O	O
by	NN	O	O
neurophils	NN	O	O
,	NN	O	O
generate	NN	O	O
neutrophil	NN	O	O
chemotactic	NN	O	O
factors	NN	O	O
and	NN	O	O
inhibit	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
E	NN	O	O
.	NN	O	O
coli	NN	O	O
;	NN	O	O
these	NN	O	O
activities	NN	O	O
were	NN	O	O
restored	NN	O	O
by	NN	O	O
purified	NN	O	O
C3	NN	O	O
.	NN	O	O

A	NN	O	O
transfusion	NN	O	O
of	NN	O	O
320	NN	O	O
ml	NN	O	O
1	NN	O	O
-	NN	O	O
hour	NN	O	O
-	NN	O	O
old	NN	O	O
normal	NN	O	O
whole	NN	O	O
blood	NN	O	O
on	NN	O	O
the	NN	O	O
fifty	NN	O	O
-	NN	O	O
second	NN	O	O
day	NN	O	O
resulted	NN	O	O
in	NN	O	O
transitory	NN	O	O
elevation	NN	O	O
of	NN	O	O
the	NN	O	O
C3	NN	O	O
level	NN	O	O
to	NN	O	O
25	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
with	NN	O	O
a	NN	O	O
fall	NN	O	O
-	NN	O	O
off	NN	O	O
(	NN	O	O
approximately	NN	O	O
2	NN	O	O
1	NN	O	O
/	NN	O	O
2	NN	O	O
%	NN	O	O
per	NN	O	O
hour	NN	O	O
)	NN	O	O
to	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
by	NN	O	O
69	NN	O	O
hours	NN	O	O
;	NN	O	O
it	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
disappearance	NN	O	O
of	NN	O	O
the	NN	O	O
skin	NN	O	B-Disease
rash	NN	O	I-Disease
and	NN	O	O
arthralgias	NN	O	B-Disease
and	NN	O	O
return	NN	O	O
to	NN	O	O
normal	NN	O	O
of	NN	O	O
the	NN	O	O
previously	NN	O	O
elevated	NN	O	O
temperature	NN	O	O
and	NN	O	O
CRP	NN	O	O
levels	NN	O	O
.	NN	O	O

C3	NN	O	O
levels	NN	O	O
in	NN	O	O
family	NN	O	O
members	NN	O	O
(	NN	O	O
seven	NN	O	O
of	NN	O	O
24	NN	O	O
half	NN	O	O
-	NN	O	O
normal	NN	O	O
)	NN	O	O
,	NN	O	O
lack	NN	O	O
of	NN	O	O
anti	NN	O	O
-	NN	O	O
C3	NN	O	O
activity	NN	O	O
,	NN	O	O
normal	NN	O	O
C3b	NN	O	O
-	NN	O	O
INA	NN	O	O
levels	NN	O	O
and	NN	O	O
a	NN	O	O
normal	NN	O	O
rate	NN	O	O
of	NN	O	O
catabolism	NN	O	O
of	NN	O	O
transfused	NN	O	O
C3	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
deficiency	NN	O	O
was	NN	O	O
inherited	NN	O	O
with	NN	O	O
autosomal	NN	O	O
codominance	NN	O	O
and	NN	O	O
involved	NN	O	O
decreased	NN	O	B-Disease
synthesis	NN	O	I-Disease
of	NN	O	I-Disease
C3	NN	O	I-Disease
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
child	NN	O	O
is	NN	O	O
a	NN	O	O
unique	NN	O	O
individual	NN	O	O
with	NN	O	O
inherited	NN	O	B-Disease
C3	NN	O	I-Disease
deficiency	NN	O	I-Disease
presenting	NN	O	O
with	NN	O	O
absence	NN	O	O
of	NN	O	O
repeated	NN	O	O
infections	NN	O	O
,	NN	O	O
whose	NN	O	O
symptoms	NN	O	O
of	NN	O	O
fever	NN	O	B-Disease
,	NN	O	O
skin	NN	O	B-Disease
rash	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
arthralgia	NN	O	B-Disease
were	NN	O	O
abated	NN	O	O
by	NN	O	O
whole	NN	O	O
blood	NN	O	O
transfusion	NN	O	O
.	NN	O	O
.	NN	O	O

Glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variants	NN	O	O
and	NN	O	O
their	NN	O	O
frequency	NN	O	O
in	NN	O	O
Guangdong	NN	O	O
,	NN	O	O
China	NN	O	O
.	NN	O	O

Erythrocyte	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
was	NN	O	O
characterized	NN	O	O
in	NN	O	O
blood	NN	O	O
samples	NN	O	O
obtained	NN	O	O
from	NN	O	O
97	NN	O	O
randomly	NN	O	O
selected	NN	O	O
males	NN	O	O
with	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
from	NN	O	O
various	NN	O	O
regions	NN	O	O
of	NN	O	O
Guangdong	NN	O	O
Province	NN	O	O
,	NN	O	O
China	NN	O	O
.	NN	O	O

Nine	NN	O	O
new	NN	O	O
variants	NN	O	O
(	NN	O	O
Gd	NN	O	O
Kaiping	NN	O	O
,	NN	O	O
Gd	NN	O	O
Boluo	NN	O	O
,	NN	O	O
Gd	NN	O	O
Huiyang	NN	O	O
,	NN	O	O
Gd	NN	O	O
Gaomin	NN	O	O
,	NN	O	O
Gd	NN	O	O
Qing	NN	O	O
-	NN	O	O
Baijiang	NN	O	O
,	NN	O	O
Gd	NN	O	O
Gaozhou	NN	O	O
,	NN	O	O
Gd	NN	O	O
Huazhou	NN	O	O
,	NN	O	O
Gd	NN	O	O
Nanhai	NN	O	O
,	NN	O	O
and	NN	O	O
Gd	NN	O	O
Guangzhou	NN	O	O
)	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
31	NN	O	O
variants	NN	O	O
found	NN	O	O
in	NN	O	O
this	NN	O	O
province	NN	O	O
,	NN	O	O
Gd	NN	O	O
Kaiping	NN	O	O
,	NN	O	O
Gd	NN	O	O
Taiwan	NN	O	O
-	NN	O	O
Hakka	NN	O	O
,	NN	O	O
Gd	NN	O	O
Haad	NN	O	O
Yai	NN	O	O
,	NN	O	O
Gd	NN	O	O
Haad	NN	O	O
Yai	NN	O	O
-	NN	O	O
like	NN	O	O
and	NN	O	O
Gd	NN	O	O
Huiyang	NN	O	O
occurred	NN	O	O
most	NN	O	O
frequently	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
each	NN	O	O
variant	NN	O	O
was	NN	O	O
calculated	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
high	NN	O	O
in	NN	O	O
this	NN	O	O
area	NN	O	O
.	NN	O	O
.	NN	O	O

Homozygous	NN	O	O
and	NN	O	O
heterozygous	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
gene	NN	O	O
in	NN	O	O
patients	NN	O	O
and	NN	O	O
carriers	NN	O	O
of	NN	O	O
severe	NN	O	B-Disease
von	NN	O	I-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Severe	NN	O	B-Disease
von	NN	O	I-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
undetectable	NN	O	O
or	NN	O	O
trace	NN	O	O
quantities	NN	O	O
of	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
in	NN	O	O
plasma	NN	O	O
and	NN	O	O
tissue	NN	O	O
stores	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
of	NN	O	O
10	NN	O	O
affected	NN	O	O
individuals	NN	O	O
from	NN	O	O
six	NN	O	O
families	NN	O	O
with	NN	O	O
this	NN	O	O
disorder	NN	O	O
using	NN	O	O
probes	NN	O	O
from	NN	O	O
the	NN	O	O
5	NN	O	O
and	NN	O	O
3	NN	O	O
ends	NN	O	O
of	NN	O	O
the	NN	O	O
vWF	NN	O	O
cDNA	NN	O	O
and	NN	O	O
with	NN	O	O
a	NN	O	O
probe	NN	O	O
extending	NN	O	O
from	NN	O	O
the	NN	O	O
5	NN	O	O
end	NN	O	O
into	NN	O	O
the	NN	O	O
central	NN	O	O
region	NN	O	O
.	NN	O	O

Southern	NN	O	O
blots	NN	O	O
of	NN	O	O
restriction	NN	O	O
endonuclease	NN	O	O
digests	NN	O	O
and	NN	O	O
gene	NN	O	O
dosage	NN	O	O
analysis	NN	O	O
measurements	NN	O	O
carried	NN	O	O
out	NN	O	O
with	NN	O	O
quantitative	NN	O	O
slot	NN	O	O
blots	NN	O	O
of	NN	O	O
undigested	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
separated	NN	O	O
these	NN	O	O
patients	NN	O	O
into	NN	O	O
three	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
group	NN	O	O
consisted	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
complete	NN	O	O
homozygous	NN	O	O
deletions	NN	O	O
of	NN	O	O
the	NN	O	O
vWF	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
four	NN	O	O
probands	NN	O	O
.	NN	O	O

Gene	NN	O	O
dosage	NN	O	O
analysis	NN	O	O
was	NN	O	O
consistent	NN	O	O
with	NN	O	O
heterozygous	NN	O	O
deletions	NN	O	O
in	NN	O	O
both	NN	O	O
of	NN	O	O
the	NN	O	O
asymptomatic	NN	O	O
parents	NN	O	O
and	NN	O	O
four	NN	O	O
asymptomatic	NN	O	O
siblings	NN	O	O
of	NN	O	O
this	NN	O	O
kindred	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0	NN	O	O
.	NN	O	O
01	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
group	NN	O	O
was	NN	O	O
comprised	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
complete	NN	O	O
heterozygous	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
vWF	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
proband	NN	O	O
and	NN	O	O
one	NN	O	O
asymptomatic	NN	O	O
parent	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
a	NN	O	O
different	NN	O	O
type	NN	O	O
of	NN	O	O
genetic	NN	O	B-Disease
abnormality	NN	O	I-Disease
was	NN	O	O
inherited	NN	O	O
from	NN	O	O
the	NN	O	O
other	NN	O	O
parent	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
patient	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
doubly	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
interacting	NN	O	O
genetic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
affecting	NN	O	O
vWF	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
third	NN	O	O
group	NN	O	O
,	NN	O	O
no	NN	O	O
gene	NN	O	O
deletions	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

Alloantibodies	NN	O	O
developed	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
kindred	NN	O	O
with	NN	O	O
homozygous	NN	O	O
deletions	NN	O	O
.	NN	O	O

These	NN	O	O
techniques	NN	O	O
should	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
identifying	NN	O	O
carriers	NN	O	O
of	NN	O	O
severe	NN	O	B-Disease
von	NN	O	I-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
and	NN	O	O
also	NN	O	O
in	NN	O	O
defining	NN	O	O
patients	NN	O	O
predictably	NN	O	O
at	NN	O	O
risk	NN	O	O
of	NN	O	O
developing	NN	O	O
alloantibodies	NN	O	O
to	NN	O	O
vWF	NN	O	O
.	NN	O	O

Sjogren	NN	O	B-Disease
-	NN	O	I-Disease
Larsson	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Impaired	NN	O	O
fatty	NN	O	O
alcohol	NN	O	O
oxidation	NN	O	O
in	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
due	NN	O	O
to	NN	O	O
deficient	NN	O	O
fatty	NN	O	O
alcohol	NN	O	O
:	NN	O	O
nicotinamide	NN	O	O
adenine	NN	O	O
dinucleotide	NN	O	O
oxidoreductase	NN	O	O
activity	NN	O	O
.	NN	O	O

Lipid	NN	O	O
metabolism	NN	O	O
was	NN	O	O
studied	NN	O	O
in	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
inherited	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
Sjogren	NN	O	B-Disease
-	NN	O	I-Disease
Larsson	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
SLS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Intact	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
incubated	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
[	NN	O	O
1	NN	O	O
-	NN	O	O
14C	NN	O	O
]	NN	O	O
palmitate	NN	O	O
accumulated	NN	O	O
more	NN	O	O
radioactive	NN	O	O
hexadecanol	NN	O	O
than	NN	O	O
did	NN	O	O
normal	NN	O	O
cells	NN	O	O
,	NN	O	O
whereas	NN	O	O
incorporation	NN	O	O
of	NN	O	O
radioactivity	NN	O	O
into	NN	O	O
other	NN	O	O
cellular	NN	O	O
lipids	NN	O	O
was	NN	O	O
unaltered	NN	O	O
.	NN	O	O

The	NN	O	O
hexadecanol	NN	O	O
content	NN	O	O
of	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
was	NN	O	O
abnormally	NN	O	O
elevated	NN	O	O
.	NN	O	O

Hexadecanol	NN	O	O
accumulation	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
increased	NN	O	O
fatty	NN	O	O
alcohol	NN	O	O
synthesis	NN	O	O
nor	NN	O	O
its	NN	O	O
deficient	NN	O	O
utilization	NN	O	O
for	NN	O	O
glycerol	NN	O	O
ether	NN	O	O
synthesis	NN	O	O
.	NN	O	O

The	NN	O	O
half	NN	O	O
-	NN	O	O
life	NN	O	O
of	NN	O	O
intracellular	NN	O	O
hexadecanol	NN	O	O
loaded	NN	O	O
into	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
was	NN	O	O
increased	NN	O	O
(	NN	O	O
70	NN	O	O
min	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
(	NN	O	O
15	NN	O	O
min	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
intact	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
showed	NN	O	O
impaired	NN	O	O
oxidation	NN	O	O
of	NN	O	O
[	NN	O	O
14C	NN	O	O
]	NN	O	O
-	NN	O	O
hexadecanol	NN	O	O
to	NN	O	O
fatty	NN	O	O
acid	NN	O	O
.	NN	O	O

Fatty	NN	O	O
alcohol	NN	O	O
NAD	NN	O	O
+	NN	O	O
oxidoreductase	NN	O	O
,	NN	O	O
the	NN	O	O
enzyme	NN	O	O
catalyzing	NN	O	O
this	NN	O	O
reaction	NN	O	O
,	NN	O	O
was	NN	O	O
deficient	NN	O	O
in	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
.	NN	O	O

Mean	NN	O	O
total	NN	O	O
activity	NN	O	O
in	NN	O	O
SLS	NN	O	B-Disease
fibroblasts	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
5	NN	O	O
)	NN	O	O
was	NN	O	O
13	NN	O	O
%	NN	O	O
of	NN	O	O
that	NN	O	O
in	NN	O	O
normal	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
and	NN	O	O
palmitoyl	NN	O	O
CoA	NN	O	O
-	NN	O	O
inhibitable	NN	O	O
activity	NN	O	O
was	NN	O	O
1	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
.	NN	O	O

Fibroblasts	NN	O	O
from	NN	O	O
two	NN	O	O
obligate	NN	O	O
SLS	NN	O	B-Disease
heterozygotes	NN	O	O
had	NN	O	O
enzyme	NN	O	O
activities	NN	O	O
intermediate	NN	O	O
between	NN	O	O
that	NN	O	O
in	NN	O	O
normal	NN	O	O
fibroblasts	NN	O	O
and	NN	O	O
individuals	NN	O	O
with	NN	O	O
SLS	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
primary	NN	O	O
defect	NN	O	O
in	NN	O	O
SLS	NN	O	B-Disease
is	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
fatty	NN	O	I-Disease
alcohol	NN	O	I-Disease
NAD	NN	O	I-Disease
+	NN	O	I-Disease
oxidoreductase	NN	O	I-Disease
.	NN	O	O

SLS	NN	O	B-Disease
represents	NN	O	O
the	NN	O	O
first	NN	O	O
inherited	NN	O	B-Disease
disorder	NN	O	I-Disease
in	NN	O	O
man	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
isolated	NN	O	O
abnormality	NN	O	B-Disease
in	NN	O	I-Disease
fatty	NN	O	I-Disease
alcohol	NN	O	I-Disease
metabolism	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Germinal	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
pedigree	NN	O	O
where	NN	O	O
the	NN	O	O
disease	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
deletion	NN	O	O
within	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
meiotic	NN	O	O
segregation	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
common	NN	O	O
female	NN	O	O
ancestor	NN	O	O
using	NN	O	O
marker	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
RFLPs	NN	O	O
)	NN	O	O
detected	NN	O	O
by	NN	O	O
probes	NN	O	O
that	NN	O	O
lie	NN	O	O
within	NN	O	O
this	NN	O	O
deletion	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
female	NN	O	O
has	NN	O	O
transmitted	NN	O	O
three	NN	O	O
distinct	NN	O	O
types	NN	O	O
of	NN	O	O
X	NN	O	O
chromosome	NN	O	O
to	NN	O	O
her	NN	O	O
offspring	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
may	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
postulating	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
arose	NN	O	O
as	NN	O	O
a	NN	O	O
postzygotic	NN	O	O
deletion	NN	O	O
within	NN	O	O
this	NN	O	O
common	NN	O	O
ancestor	NN	O	O
,	NN	O	O
who	NN	O	O
was	NN	O	O
consequently	NN	O	O
germinally	NN	O	O
mosaic	NN	O	O
.	NN	O	O
.	NN	O	O

Nebulin	NN	O	O
seen	NN	O	O
in	NN	O	O
DMD	NN	O	B-Disease
males	NN	O	O
including	NN	O	O
one	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
large	NN	O	O
DNA	NN	O	O
deletion	NN	O	O
encompassing	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
nebulin	NN	O	O
in	NN	O	O
a	NN	O	O
muscle	NN	O	O
specimen	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
large	NN	O	O
deletion	NN	O	O
precludes	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
.	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
muscle	NN	O	O
and	NN	O	O
brain	NN	O	O
.	NN	O	O

Complementary	NN	O	O
DNA	NN	O	O
clones	NN	O	O
were	NN	O	O
isolated	NN	O	O
that	NN	O	O
represent	NN	O	O
the	NN	O	O
5	NN	O	O
terminal	NN	O	O
2	NN	O	O
.	NN	O	O

5	NN	O	O
kilobases	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
Dmd	NN	O	O
)	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
(	NN	O	O
mRNA	NN	O	O
)	NN	O	O
.	NN	O	O

Mouse	NN	O	O
Dmd	NN	O	O
mRNA	NN	O	O
was	NN	O	O
detectable	NN	O	O
in	NN	O	O
skeletal	NN	O	O
and	NN	O	O
cardiac	NN	O	O
muscle	NN	O	O
and	NN	O	O
at	NN	O	O
a	NN	O	O
level	NN	O	O
approximately	NN	O	O
90	NN	O	O
percent	NN	O	O
lower	NN	O	O
in	NN	O	O
brain	NN	O	O
.	NN	O	O

Dmd	NN	O	O
mRNA	NN	O	O
is	NN	O	O
also	NN	O	O
present	NN	O	O
,	NN	O	O
but	NN	O	O
at	NN	O	O
much	NN	O	O
lower	NN	O	O
than	NN	O	O
normal	NN	O	O
levels	NN	O	O
,	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
muscle	NN	O	O
and	NN	O	O
brain	NN	O	O
of	NN	O	O
three	NN	O	O
different	NN	O	O
strains	NN	O	O
of	NN	O	O
dystrophic	NN	O	B-Disease
mdx	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
Dmd	NN	O	O
mRNA	NN	O	O
in	NN	O	O
brain	NN	O	O
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
relation	NN	O	O
between	NN	O	O
human	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
found	NN	O	O
in	NN	O	O
some	NN	O	O
DMD	NN	O	B-Disease
males	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
also	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
mdx	NN	O	O
mutations	NN	O	O
are	NN	O	O
allelic	NN	O	O
variants	NN	O	O
of	NN	O	O
mouse	NN	O	O
Dmd	NN	O	O
gene	NN	O	O
mutations	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
incidence	NN	O	O
of	NN	O	O
hematologic	NN	O	B-Disease
malignancy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
evaluated	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
a	NN	O	O
negative	NN	O	O
association	NN	O	O
between	NN	O	O
glucose	NN	O	B-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
cancer	NN	O	B-Disease
in	NN	O	O
a	NN	O	O
cohort	NN	O	O
of	NN	O	O
481	NN	O	O
Sardinian	NN	O	O
males	NN	O	O
with	NN	O	O
hematological	NN	O	B-Disease
malignancies	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
not	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
incidence	NN	O	O
in	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
16	NN	O	O
,	NN	O	O
219	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
conclusion	NN	O	O
resulted	NN	O	O
from	NN	O	O
the	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
GdB	NN	O	O
gene	NN	O	O
in	NN	O	O
23	NN	O	O
heterozygous	NN	O	O
women	NN	O	O
having	NN	O	O
a	NN	O	O
clonal	NN	O	O
hematologic	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
control	NN	O	O
group	NN	O	O
of	NN	O	O
37	NN	O	O
healthy	NN	O	O
heterozygotes	NN	O	O
.	NN	O	O

Therefore	NN	O	O
at	NN	O	O
present	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
evidence	NN	O	O
that	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
has	NN	O	O
a	NN	O	O
protective	NN	O	O
effect	NN	O	O
against	NN	O	O
development	NN	O	O
of	NN	O	O
hematologic	NN	O	B-Disease
neoplasms	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Hepatoblastoma	NN	O	B-Disease
,	NN	O	O
pigmented	NN	O	B-Disease
ocular	NN	O	I-Disease
fundus	NN	O	I-Disease
lesions	NN	O	I-Disease
and	NN	O	O
jaw	NN	O	B-Disease
lesions	NN	O	I-Disease
in	NN	O	O
Gardner	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Hepatoblastoma	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	O
neoplasm	NN	O	B-Disease
of	NN	O	O
infants	NN	O	O
and	NN	O	O
children	NN	O	O
only	NN	O	O
recently	NN	O	O
documented	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
adenomatous	NN	O	I-Disease
polyposis	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
colon	NN	O	I-Disease
[	NN	O	O
Kingston	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1983	NN	O	O
]	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
four	NN	O	O
children	NN	O	O
with	NN	O	O
hepatoblastoma	NN	O	B-Disease
from	NN	O	O
four	NN	O	O
unrelated	NN	O	O
families	NN	O	O
with	NN	O	O
Gardner	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
GS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

One	NN	O	O
child	NN	O	O
,	NN	O	O
now	NN	O	O
19	NN	O	O
years	NN	O	O
old	NN	O	O
,	NN	O	O
survived	NN	O	O
after	NN	O	O
a	NN	O	O
resection	NN	O	O
of	NN	O	O
a	NN	O	O
hepatoblastoma	NN	O	B-Disease
in	NN	O	O
infancy	NN	O	O
and	NN	O	O
recently	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
GS	NN	O	B-Disease
.	NN	O	O

He	NN	O	O
has	NN	O	O
an	NN	O	O
associated	NN	O	O
odontoma	NN	O	B-Disease
and	NN	O	O
pigmented	NN	O	B-Disease
ocular	NN	O	I-Disease
fundus	NN	O	I-Disease
lesions	NN	O	I-Disease
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
clinical	NN	O	O
markers	NN	O	O
of	NN	O	O
GS	NN	O	B-Disease
.	NN	O	O

Many	NN	O	O
individuals	NN	O	O
in	NN	O	O
these	NN	O	O
four	NN	O	O
GS	NN	O	B-Disease
families	NN	O	O
,	NN	O	O
both	NN	O	O
affected	NN	O	O
and	NN	O	O
at	NN	O	O
risk	NN	O	O
,	NN	O	O
have	NN	O	O
osteomatous	NN	O	B-Disease
jaw	NN	O	I-Disease
lesions	NN	O	I-Disease
and	NN	O	O
pigmented	NN	O	B-Disease
ocular	NN	O	I-Disease
fundus	NN	O	I-Disease
lesions	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
search	NN	O	O
for	NN	O	O
colonic	NN	O	B-Disease
polyps	NN	O	I-Disease
should	NN	O	O
be	NN	O	O
made	NN	O	O
in	NN	O	O
families	NN	O	O
of	NN	O	O
infants	NN	O	O
and	NN	O	O
children	NN	O	O
with	NN	O	O
hepatoblastoma	NN	O	B-Disease
.	NN	O	O

If	NN	O	O
the	NN	O	O
child	NN	O	O
survives	NN	O	O
,	NN	O	O
he	NN	O	O
or	NN	O	O
she	NN	O	O
should	NN	O	O
be	NN	O	O
monitored	NN	O	O
for	NN	O	O
the	NN	O	O
later	NN	O	O
appearance	NN	O	O
of	NN	O	O
colonic	NN	O	B-Disease
polyps	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
finding	NN	O	O
of	NN	O	O
jaw	NN	O	B-Disease
lesions	NN	O	I-Disease
and	NN	O	O
/	NN	O	O
or	NN	O	O
pigmented	NN	O	B-Disease
ocular	NN	O	I-Disease
fundus	NN	O	I-Disease
lesions	NN	O	I-Disease
in	NN	O	O
relatives	NN	O	O
at	NN	O	O
risk	NN	O	O
are	NN	O	O
indications	NN	O	O
of	NN	O	O
the	NN	O	O
possible	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
GS	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
an	NN	O	O
altered	NN	O	O
splice	NN	O	O
site	NN	O	O
in	NN	O	O
Ashkenazi	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
resulting	NN	O	O
from	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
enzyme	NN	O	O
,	NN	O	O
beta	NN	O	O
-	NN	O	O
N	NN	O	O
-	NN	O	O
acetylhexosaminidase	NN	O	O
A	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
relatively	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
carriers	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
27	NN	O	O
)	NN	O	O
of	NN	O	O
a	NN	O	O
lethal	NN	O	O
,	NN	O	O
infantile	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
population	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
evident	NN	O	O
whether	NN	O	O
this	NN	O	O
has	NN	O	O
resulted	NN	O	O
from	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
and	NN	O	O
random	NN	O	O
genetic	NN	O	O
drift	NN	O	O
or	NN	O	O
from	NN	O	O
a	NN	O	O
selective	NN	O	O
advantage	NN	O	O
of	NN	O	O
heterozygotes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
single	NN	O	O
-	NN	O	O
base	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
cloned	NN	O	O
fragment	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
from	NN	O	O
an	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
patient	NN	O	O
.	NN	O	O

This	NN	O	O
change	NN	O	O
,	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
a	NN	O	O
C	NN	O	O
for	NN	O	O
G	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
nucleotide	NN	O	O
of	NN	O	O
intron	NN	O	O
12	NN	O	O
is	NN	O	O
expected	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
defective	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
.	NN	O	O

A	NN	O	O
test	NN	O	O
for	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
based	NN	O	O
on	NN	O	O
amplification	NN	O	O
of	NN	O	O
DNA	NN	O	O
by	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
rection	NN	O	O
and	NN	O	O
cleavage	NN	O	O
of	NN	O	O
a	NN	O	O
DdeI	NN	O	O
restriction	NN	O	O
site	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
mutation	NN	O	O
revealed	NN	O	O
that	NN	O	O
this	NN	O	O
case	NN	O	O
and	NN	O	O
two	NN	O	O
other	NN	O	O
cases	NN	O	O
of	NN	O	O
the	NN	O	O
Ashkenazi	NN	O	O
,	NN	O	O
infantile	NN	O	O
form	NN	O	O
of	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
are	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
two	NN	O	O
different	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
occurrence	NN	O	O
of	NN	O	O
multiple	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
warrants	NN	O	O
further	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
selective	NN	O	O
advantage	NN	O	O
hypothesis	NN	O	O
.	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
glucose	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Mus	NN	O	O
musculus	NN	O	O
.	NN	O	O

A	NN	O	O
mouse	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
glucose	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
recovered	NN	O	O
in	NN	O	O
offspring	NN	O	O
of	NN	O	O
1	NN	O	O
-	NN	O	O
ethyl	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
nitrosourea	NN	O	O
-	NN	O	O
treated	NN	O	O
male	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
alteration	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
blood	NN	O	O
but	NN	O	O
can	NN	O	O
also	NN	O	O
be	NN	O	O
observed	NN	O	O
in	NN	O	O
other	NN	O	O
tissue	NN	O	O
extracts	NN	O	O
.	NN	O	O

Hemizygous	NN	O	O
,	NN	O	O
heterozygous	NN	O	O
,	NN	O	O
and	NN	O	O
homozygous	NN	O	O
mutants	NN	O	O
have	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
about	NN	O	O
15	NN	O	O
,	NN	O	O
60	NN	O	O
,	NN	O	O
and	NN	O	O
15	NN	O	O
%	NN	O	O
G6PD	NN	O	O
remaining	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
blood	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
wild	NN	O	O
type	NN	O	O
.	NN	O	O

Erythrocyte	NN	O	O
indices	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
differences	NN	O	O
between	NN	O	O
mutants	NN	O	O
and	NN	O	O
wild	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
migration	NN	O	O
,	NN	O	O
the	NN	O	O
isoelectric	NN	O	O
point	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
thermal	NN	O	O
stability	NN	O	O
.	NN	O	O

Kinetic	NN	O	O
properties	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
Km	NN	O	O
for	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
or	NN	O	O
for	NN	O	O
NADP	NN	O	O
and	NN	O	O
the	NN	O	O
relative	NN	O	O
utilization	NN	O	O
of	NN	O	O
substrate	NN	O	O
analogues	NN	O	O
,	NN	O	O
showed	NN	O	O
no	NN	O	O
differences	NN	O	O
between	NN	O	O
wild	NN	O	O
types	NN	O	O
and	NN	O	O
mutants	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
the	NN	O	O
relative	NN	O	O
utilization	NN	O	O
of	NN	O	O
deamino	NN	O	O
-	NN	O	O
NADP	NN	O	O
which	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
mutants	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
presently	NN	O	O
the	NN	O	O
only	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
G6PD	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
humans	NN	O	O
.	NN	O	O
.	NN	O	O

Diverse	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
gene	NN	O	O
cause	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
mild	NN	O	O
or	NN	O	O
severe	NN	O	O
hemolytic	NN	O	B-Disease
anemia	NN	O	I-Disease
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
;	NN	O	I-Disease
EC	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
49	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
common	NN	O	O
genetic	NN	O	B-Disease
abnormality	NN	O	I-Disease
affecting	NN	O	O
an	NN	O	O
estimated	NN	O	O
400	NN	O	O
million	NN	O	O
people	NN	O	O
worldwide	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
analyses	NN	O	O
have	NN	O	O
identified	NN	O	O
many	NN	O	O
variants	NN	O	O
exhibiting	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
phenotypes	NN	O	O
,	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
from	NN	O	O
the	NN	O	O
hematological	NN	O	O
point	NN	O	O
of	NN	O	O
view	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
until	NN	O	O
now	NN	O	O
,	NN	O	O
their	NN	O	O
precise	NN	O	O
molecular	NN	O	O
basis	NN	O	O
has	NN	O	O
remained	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
seven	NN	O	O
mutant	NN	O	O
G6PD	NN	O	O
alleles	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
nondeficient	NN	O	O
polymorphic	NN	O	O
African	NN	O	O
variant	NN	O	O
G6PD	NN	O	O
A	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
a	NN	O	O
single	NN	O	O
point	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
six	NN	O	O
mutants	NN	O	O
investigated	NN	O	O
were	NN	O	O
all	NN	O	O
associated	NN	O	O
with	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
commonest	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
favism	NN	O	B-Disease
among	NN	O	O
other	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
,	NN	O	O
a	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
replacement	NN	O	O
was	NN	O	O
found	NN	O	O
(	NN	O	O
serine	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
phenylalanine	NN	O	O
)	NN	O	O
it	NN	O	O
must	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
decreased	NN	O	O
stability	NN	O	O
and	NN	O	O
the	NN	O	O
reduced	NN	O	O
catalytic	NN	O	O
efficiency	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
.	NN	O	O

Single	NN	O	O
point	NN	O	O
mutations	NN	O	O
were	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
G6PD	NN	O	O
Metaponto	NN	O	O
(	NN	O	O
Southern	NN	O	O
Italy	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
G6PD	NN	O	O
Ilesha	NN	O	O
(	NN	O	O
Nigeria	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
asymptomatic	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
G6PD	NN	O	O
Chatham	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
an	NN	O	O
Indian	NN	O	O
boy	NN	O	O
with	NN	O	O
neonatal	NN	O	B-Disease
jaundice	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
G6PD	NN	O	O
"	NN	O	O
Matera	NN	O	O
,	NN	O	O
"	NN	O	O
which	NN	O	O
is	NN	O	O
now	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
same	NN	O	O
as	NN	O	O
G6PD	NN	O	O
A	NN	O	O
-	NN	O	O
,	NN	O	O
two	NN	O	O
separate	NN	O	O
point	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
same	NN	O	O
as	NN	O	O
in	NN	O	O
G6PD	NN	O	O
A	NN	O	O
.	NN	O	O

In	NN	O	O
G6PD	NN	O	O
Santiago	NN	O	O
,	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
mutation	NN	O	O
(	NN	O	O
glycine	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
arginine	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
severe	NN	O	O
chronic	NN	O	O
hemolytic	NN	O	B-Disease
anemia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
observed	NN	O	O
show	NN	O	O
a	NN	O	O
striking	NN	O	O
predominance	NN	O	O
of	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
T	NN	O	O
transitions	NN	O	O
,	NN	O	O
with	NN	O	O
CG	NN	O	O
doublets	NN	O	O
involved	NN	O	O
in	NN	O	O
four	NN	O	O
of	NN	O	O
seven	NN	O	O
cases	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
diverse	NN	O	O
point	NN	O	O
mutations	NN	O	O
may	NN	O	O
account	NN	O	O
largely	NN	O	O
for	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Tight	NN	O	O
linkage	NN	O	O
between	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
apolipoprotein	NN	O	O
E	NN	O	O
genes	NN	O	O
revealed	NN	O	O
with	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

In	NN	O	O
16	NN	O	O
families	NN	O	O
with	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
based	NN	O	O
on	NN	O	O
use	NN	O	O
of	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
oligonucleotides	NN	O	O
has	NN	O	O
been	NN	O	O
employed	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
linkage	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
E	NN	O	O
(	NN	O	O
APOE	NN	O	O
)	NN	O	O
gene	NN	O	O
and	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

Synthetic	NN	O	O
oligonucleotides	NN	O	O
,	NN	O	O
designed	NN	O	O
to	NN	O	O
discriminate	NN	O	O
between	NN	O	O
APOE	NN	O	O
alleles	NN	O	O
epsilon	NN	O	O
3	NN	O	O
and	NN	O	O
epsilon	NN	O	O
4	NN	O	O
,	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
distinguish	NN	O	O
heterozygous	NN	O	O
carriers	NN	O	O
in	NN	O	O
a	NN	O	O
hybridization	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
families	NN	O	O
,	NN	O	O
the	NN	O	O
relevant	NN	O	O
segment	NN	O	O
of	NN	O	O
the	NN	O	O
APOE	NN	O	O
gene	NN	O	O
was	NN	O	O
enzymatically	NN	O	O
amplified	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
method	NN	O	O
.	NN	O	O

For	NN	O	O
DM	NN	O	O
and	NN	O	O
APOE	NN	O	O
,	NN	O	O
a	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
(	NN	O	O
zmax	NN	O	O
of	NN	O	O
7	NN	O	O
.	NN	O	O

47	NN	O	O
was	NN	O	O
obtained	NN	O	O
at	NN	O	O
a	NN	O	O
recombination	NN	O	O
frequency	NN	O	O
(	NN	O	O
theta	NN	O	O
)	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

047	NN	O	O
(	NN	O	O
male	NN	O	O
theta	NN	O	O
=	NN	O	O
female	NN	O	O
theta	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
recombination	NN	O	O
(	NN	O	O
maximum	NN	O	O
lod	NN	O	O
score	NN	O	O
of	NN	O	O
5	NN	O	O
.	NN	O	O
61	NN	O	O
at	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
0	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
APOE	NN	O	O
and	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
CII	NN	O	O
(	NN	O	O
APOC2	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
APOC2	NN	O	O
,	NN	O	O
APOE	NN	O	O
is	NN	O	O
a	NN	O	O
useful	NN	O	O
marker	NN	O	O
for	NN	O	O
presymptomatic	NN	O	O
DM	NN	O	B-Disease
diagnosis	NN	O	O
.	NN	O	O

Regional	NN	O	O
localization	NN	O	O
of	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
loci	NN	O	O
on	NN	O	O
the	NN	O	O
proximal	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
using	NN	O	O
deletions	NN	O	O
associated	NN	O	O
with	NN	O	O
choroideremia	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
two	NN	O	O
unrelated	NN	O	O
families	NN	O	O
,	NN	O	O
males	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
who	NN	O	O
suffer	NN	O	O
from	NN	O	O
choroideremia	NN	O	B-Disease
and	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
have	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
on	NN	O	O
the	NN	O	O
proximal	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

By	NN	O	O
high	NN	O	O
-	NN	O	O
resolution	NN	O	O
banding	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
deletion	NN	O	O
chromosomes	NN	O	O
as	NN	O	O
del	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
(	NN	O	O
q21	NN	O	O
.	NN	O	O
1	NN	O	O
-	NN	O	O
q21	NN	O	O
1	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O
33	NN	O	O
)	NN	O	O
and	NN	O	O
del	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
(	NN	O	O
q21	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
q21	NN	O	O
2	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O
31	NN	O	O
)	NN	O	O
respectively	NN	O	O
.	NN	O	O

By	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
we	NN	O	O
have	NN	O	O
mapped	NN	O	O
ten	NN	O	O
different	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
loci	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
and	NN	O	O
the	NN	O	O
choroideremia	NN	O	B-Disease
locus	NN	O	O
TCD	NN	O	O
.	NN	O	O

One	NN	O	O
probe	NN	O	O
,	NN	O	O
p31	NN	O	O
,	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
cover	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
the	NN	O	O
smallest	NN	O	O
deletion	NN	O	O
.	NN	O	O

The	NN	O	O
following	NN	O	O
order	NN	O	O
of	NN	O	O
the	NN	O	O
loci	NN	O	O
was	NN	O	O
suggested	NN	O	O
by	NN	O	O
deletion	NN	O	O
mapping	NN	O	O
cen	NN	O	O
-	NN	O	O
DXS106	NN	O	O
-	NN	O	O
DXS72	NN	O	O
-	NN	O	O
TCD	NN	O	O
-	NN	O	O
(	NN	O	O
DXYS1	NN	O	O
/	NN	O	O
DXYS23	NN	O	O
/	NN	O	O
DXYS5	NN	O	O
)	NN	O	O
-	NN	O	O
DXYS2	NN	O	O
-	NN	O	O
(	NN	O	O
DXYS12	NN	O	O
/	NN	O	O
DXS3	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
DXS17	NN	O	O
/	NN	O	O
DXS101	NN	O	O
)	NN	O	O
-	NN	O	O
Xqter	NN	O	O
.	NN	O	O

Retroviral	NN	O	O
-	NN	O	O
mediated	NN	O	O
gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
human	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
into	NN	O	O
NIH	NN	O	O
3T3	NN	O	O
and	NN	O	O
hepatoma	NN	O	B-Disease
cells	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
hepatic	NN	O	I-Disease
enzyme	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
human	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
sequence	NN	O	O
has	NN	O	O
been	NN	O	O
inserted	NN	O	O
into	NN	O	O
pzip	NN	O	O
-	NN	O	O
neoSV	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
retroviral	NN	O	O
vector	NN	O	O
containing	NN	O	O
the	NN	O	O
bacterial	NN	O	O
neo	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
recombinant	NN	O	O
has	NN	O	O
been	NN	O	O
transfected	NN	O	O
into	NN	O	O
psi	NN	O	O
2	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
provide	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
retroviral	NN	O	O
capsid	NN	O	O
.	NN	O	O

Recombinant	NN	O	O
virus	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
of	NN	O	O
the	NN	O	O
transfected	NN	O	O
psi	NN	O	O
2	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
transmitting	NN	O	O
the	NN	O	O
human	NN	O	O
PAH	NN	O	O
gene	NN	O	O
into	NN	O	O
mouse	NN	O	O
NIH	NN	O	O
3T3	NN	O	O
cells	NN	O	O
by	NN	O	O
infection	NN	O	O
leading	NN	O	O
to	NN	O	O
stable	NN	O	O
incorporation	NN	O	O
of	NN	O	O
the	NN	O	O
recombinant	NN	O	O
provirus	NN	O	O
.	NN	O	O

Infected	NN	O	O
cells	NN	O	O
express	NN	O	O
PAH	NN	O	O
mRNA	NN	O	O
,	NN	O	O
immunoreactive	NN	O	O
PAH	NN	O	O
protein	NN	O	O
,	NN	O	O
and	NN	O	O
exhibit	NN	O	O
pterin	NN	O	O
-	NN	O	O
dependent	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
recombinant	NN	O	O
virus	NN	O	O
is	NN	O	O
also	NN	O	O
capable	NN	O	O
of	NN	O	O
infecting	NN	O	O
a	NN	O	O
mouse	NN	O	O
hepatoma	NN	O	B-Disease
cell	NN	O	O
line	NN	O	O
that	NN	O	O
does	NN	O	O
not	NN	O	O
normally	NN	O	O
synthesize	NN	O	O
PAH	NN	O	O
.	NN	O	O

PAH	NN	O	O
activity	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
extracts	NN	O	O
and	NN	O	O
the	NN	O	O
entire	NN	O	O
hydroxylation	NN	O	O
system	NN	O	O
is	NN	O	O
reconstituted	NN	O	O
in	NN	O	O
the	NN	O	O
hepatoma	NN	O	B-Disease
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
the	NN	O	O
recombinant	NN	O	O
viruses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
recombinant	NN	O	O
viruses	NN	O	O
containing	NN	O	O
human	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
provide	NN	O	O
a	NN	O	O
means	NN	O	O
for	NN	O	O
introducing	NN	O	O
functional	NN	O	O
PAH	NN	O	O
into	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
of	NN	O	O
hepatic	NN	O	O
origin	NN	O	O
and	NN	O	O
can	NN	O	O
potentially	NN	O	O
be	NN	O	O
introduced	NN	O	O
into	NN	O	O
whole	NN	O	O
animals	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
somatic	NN	O	O
gene	NN	O	O
therapy	NN	O	O
for	NN	O	O
PKU	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Estimation	NN	O	O
of	NN	O	O
the	NN	O	O
male	NN	O	O
to	NN	O	O
female	NN	O	O
ratio	NN	O	O
of	NN	O	O
mutation	NN	O	O
rates	NN	O	O
from	NN	O	O
the	NN	O	O
segregation	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
chromosomal	NN	O	O
DNA	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
procedure	NN	O	O
is	NN	O	O
presented	NN	O	O
to	NN	O	O
estimate	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
male	NN	O	O
to	NN	O	O
female	NN	O	O
mutation	NN	O	O
rates	NN	O	O
for	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

X	NN	O	O
-	NN	O	O
specific	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
are	NN	O	O
used	NN	O	O
to	NN	O	O
establish	NN	O	O
DNA	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
three	NN	O	O
-	NN	O	O
generation	NN	O	O
DMD	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

From	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
who	NN	O	O
have	NN	O	O
inherited	NN	O	O
their	NN	O	O
maternal	NN	O	O
grandfathers	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
mutation	NN	O	O
rates	NN	O	O
can	NN	O	O
be	NN	O	O
calculated	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
classical	NN	O	O
methods	NN	O	O
,	NN	O	O
the	NN	O	O
proposed	NN	O	O
procedure	NN	O	O
is	NN	O	O
not	NN	O	O
restricted	NN	O	O
to	NN	O	O
sporadic	NN	O	O
or	NN	O	O
familiar	NN	O	O
cases	NN	O	O
nor	NN	O	O
is	NN	O	O
any	NN	O	O
information	NN	O	O
on	NN	O	O
the	NN	O	O
carrier	NN	O	O
status	NN	O	O
of	NN	O	O
female	NN	O	O
relatives	NN	O	O
required	NN	O	O
.	NN	O	O
.	NN	O	O

Deletions	NN	O	O
of	NN	O	O
a	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
in	NN	O	O
retinoblastomas	NN	O	B-Disease
and	NN	O	O
mesenchymal	NN	O	B-Disease
tumors	NN	O	I-Disease
:	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
and	NN	O	O
its	NN	O	O
encoded	NN	O	O
protein	NN	O	O
.	NN	O	O

Retinoblastoma	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
childhood	NN	O	B-Disease
tumor	NN	O	I-Disease
that	NN	O	O
can	NN	O	O
arise	NN	O	O
because	NN	O	O
of	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
acquired	NN	O	O
as	NN	O	O
somatic	NN	O	O
or	NN	O	O
germinal	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
allele	NN	O	O
can	NN	O	O
be	NN	O	O
carried	NN	O	O
in	NN	O	O
the	NN	O	O
germ	NN	O	O
line	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
creating	NN	O	O
these	NN	O	O
alleles	NN	O	O
act	NN	O	O
by	NN	O	O
inactivating	NN	O	O
copies	NN	O	O
of	NN	O	O
a	NN	O	O
recessive	NN	O	O
oncogene	NN	O	O
located	NN	O	O
within	NN	O	O
band	NN	O	O
q14	NN	O	O
of	NN	O	O
chromosome	NN	O	O
13	NN	O	O
and	NN	O	O
termed	NN	O	O
the	NN	O	O
RB1	NN	O	O
locus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
reported	NN	O	O
isolation	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
fragment	NN	O	O
that	NN	O	O
recognizes	NN	O	O
chromosomal	NN	O	O
sequences	NN	O	O
possessing	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
attributes	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-Disease
gene	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
RB1	NN	O	O
locus	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
this	NN	O	O
segment	NN	O	O
is	NN	O	O
additionally	NN	O	O
the	NN	O	O
target	NN	O	O
of	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
mesenchymal	NN	O	B-Disease
tumors	NN	O	I-Disease
among	NN	O	O
patients	NN	O	O
having	NN	O	O
no	NN	O	O
apparent	NN	O	O
predisposition	NN	O	O
to	NN	O	O
retinoblastoma	NN	O	B-Disease
and	NN	O	O
no	NN	O	O
previous	NN	O	O
evidence	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
tumors	NN	O	B-Disease
provide	NN	O	O
additional	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
cloned	NN	O	O
sequences	NN	O	O
are	NN	O	O
representative	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
a	NN	O	O
frequent	NN	O	O
target	NN	O	O
of	NN	O	O
inactivation	NN	O	O
during	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
this	NN	O	O
cDNA	NN	O	O
provides	NN	O	O
little	NN	O	O
insight	NN	O	O
into	NN	O	O
its	NN	O	O
normal	NN	O	O
functional	NN	O	O
role	NN	O	O
.	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
with	NN	O	O
growth	NN	O	O
hormone	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

A	NN	O	O
controlled	NN	O	O
,	NN	O	O
double	NN	O	O
-	NN	O	O
blind	NN	O	O
therapeutic	NN	O	O
trial	NN	O	O
with	NN	O	O
the	NN	O	O
drug	NN	O	O
mazindol	NN	O	O
,	NN	O	O
a	NN	O	O
growth	NN	O	O
hormone	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
a	NN	O	O
pair	NN	O	O
of	NN	O	O
7	NN	O	O
1	NN	O	O
/	NN	O	O
2	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
monozygotic	NN	O	O
twins	NN	O	O
,	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
rationale	NN	O	O
for	NN	O	O
this	NN	O	O
trial	NN	O	O
was	NN	O	O
based	NN	O	O
on	NN	O	O
a	NN	O	O
patient	NN	O	O
(	NN	O	O
reported	NN	O	O
previously	NN	O	O
)	NN	O	O
affected	NN	O	O
simultaneously	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
growth	NN	O	B-Disease
hormone	NN	O	I-Disease
(	NN	O	I-Disease
GH	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
who	NN	O	O
is	NN	O	O
showing	NN	O	O
a	NN	O	O
benign	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
dystrophic	NN	O	B-Disease
process	NN	O	I-Disease
and	NN	O	O
is	NN	O	O
still	NN	O	O
walking	NN	O	O
at	NN	O	O
18	NN	O	O
years	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
twins	NN	O	O
received	NN	O	O
2	NN	O	O
mg	NN	O	O
of	NN	O	O
mazindol	NN	O	O
daily	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
other	NN	O	O
received	NN	O	O
a	NN	O	O
placebo	NN	O	O
.	NN	O	O

The	NN	O	O
assessment	NN	O	O
,	NN	O	O
repeated	NN	O	O
every	NN	O	O
2	NN	O	O
months	NN	O	O
,	NN	O	O
included	NN	O	O
weight	NN	O	O
and	NN	O	O
height	NN	O	O
measurements	NN	O	O
,	NN	O	O
functional	NN	O	O
and	NN	O	O
motor	NN	O	O
ability	NN	O	O
tests	NN	O	O
,	NN	O	O
ergometry	NN	O	O
and	NN	O	O
determinations	NN	O	O
of	NN	O	O
serum	NN	O	O
enzymes	NN	O	O
and	NN	O	O
GH	NN	O	O
levels	NN	O	O
.	NN	O	O

After	NN	O	O
one	NN	O	O
year	NN	O	O
of	NN	O	O
trial	NN	O	O
the	NN	O	O
code	NN	O	O
was	NN	O	O
broken	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
seen	NN	O	O
that	NN	O	O
the	NN	O	O
twin	NN	O	O
under	NN	O	O
placebo	NN	O	O
treatment	NN	O	O
was	NN	O	O
strikingly	NN	O	O
worse	NN	O	O
than	NN	O	O
his	NN	O	O
brother	NN	O	O
,	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
whose	NN	O	O
condition	NN	O	O
was	NN	O	O
practically	NN	O	O
arrested	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
treatment	NN	O	O
with	NN	O	O
a	NN	O	O
GH	NN	O	O
inhibitor	NN	O	O
is	NN	O	O
beneficial	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
and	NN	O	O
localization	NN	O	O
of	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
locus	NN	O	O
by	NN	O	O
ribonuclease	NN	O	O
A	NN	O	O
cleavage	NN	O	O
.	NN	O	O

Many	NN	O	O
mutations	NN	O	O
leading	NN	O	O
to	NN	O	O
human	NN	O	O
disease	NN	O	O
are	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
single	NN	O	O
DNA	NN	O	O
base	NN	O	O
pair	NN	O	O
changes	NN	O	O
that	NN	O	O
cannot	NN	O	O
be	NN	O	O
identified	NN	O	O
by	NN	O	O
Southern	NN	O	O
analysis	NN	O	O
.	NN	O	O

This	NN	O	O
has	NN	O	O
prompted	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
alternative	NN	O	O
assays	NN	O	O
for	NN	O	O
point	NN	O	O
mutation	NN	O	O
detection	NN	O	O
.	NN	O	O

The	NN	O	O
recently	NN	O	O
described	NN	O	O
ribonuclease	NN	O	O
A	NN	O	O
cleavage	NN	O	O
procedure	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
polyuridylic	NN	O	O
acid	NN	O	O
-	NN	O	O
paper	NN	O	O
affinity	NN	O	O
chromatography	NN	O	O
step	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
mutational	NN	O	O
lesions	NN	O	O
in	NN	O	O
the	NN	O	O
hypoxanthine	NN	O	O
phosphoribosyltransferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
)	NN	O	O
messenger	NN	O	O
RNAs	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Distinctive	NN	O	O
ribonuclease	NN	O	O
A	NN	O	O
cleavage	NN	O	O
patterns	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
from	NN	O	O
5	NN	O	O
of	NN	O	O
14	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patients	NN	O	O
who	NN	O	O
were	NN	O	O
chosen	NN	O	O
because	NN	O	O
no	NN	O	O
HPRT	NN	O	O
Southern	NN	O	O
or	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
pattern	NN	O	O
changes	NN	O	O
had	NN	O	O
been	NN	O	O
found	NN	O	O
.	NN	O	O

This	NN	O	O
approach	NN	O	O
now	NN	O	O
allows	NN	O	O
HPRT	NN	O	O
mutation	NN	O	O
detection	NN	O	O
in	NN	O	O
50	NN	O	O
percent	NN	O	O
of	NN	O	O
the	NN	O	O
cases	NN	O	O
of	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
polyuridylic	NN	O	O
acid	NN	O	O
-	NN	O	O
paper	NN	O	O
affinity	NN	O	O
procedure	NN	O	O
provides	NN	O	O
a	NN	O	O
general	NN	O	O
method	NN	O	O
for	NN	O	O
analysis	NN	O	O
of	NN	O	O
low	NN	O	O
abundance	NN	O	O
messenger	NN	O	O
RNAs	NN	O	O
.	NN	O	O
.	NN	O	O

Two	NN	O	O
new	NN	O	O
variants	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
associated	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
non	NN	O	I-Disease
-	NN	O	I-Disease
spherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
:	NN	O	O
G6PD	NN	O	O
Wayne	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Huron	NN	O	O
.	NN	O	O

Two	NN	O	O
new	NN	O	O
deficient	NN	O	O
variants	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
causing	NN	O	O
hereditary	NN	O	B-Disease
nonspherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
(	NN	O	O
HNSHA	NN	O	B-Disease
)	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
are	NN	O	O
unique	NN	O	O
and	NN	O	O
they	NN	O	O
have	NN	O	O
been	NN	O	O
named	NN	O	O
G6PD	NN	O	O
Wayne	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Huron	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
G6PD	NN	O	O
Wayne	NN	O	O
underwent	NN	O	O
splenectomy	NN	O	O
and	NN	O	O
no	NN	O	O
objective	NN	O	O
improvement	NN	O	O
was	NN	O	O
noted	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
with	NN	O	O
G6PD	NN	O	O
Huron	NN	O	O
were	NN	O	O
under	NN	O	O
medical	NN	O	O
observation	NN	O	O
for	NN	O	O
a	NN	O	O
considerable	NN	O	O
period	NN	O	O
of	NN	O	O
time	NN	O	O
without	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
being	NN	O	O
entertained	NN	O	O
because	NN	O	O
the	NN	O	O
family	NN	O	O
was	NN	O	O
of	NN	O	O
Northern	NN	O	O
European	NN	O	O
origin	NN	O	O
.	NN	O	O

Since	NN	O	O
sporadic	NN	O	O
variants	NN	O	O
of	NN	O	O
G6PD	NN	O	O
causing	NN	O	O
HNSHA	NN	O	B-Disease
show	NN	O	O
no	NN	O	O
special	NN	O	O
racial	NN	O	O
predilection	NN	O	O
,	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
should	NN	O	O
always	NN	O	O
be	NN	O	O
considered	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
this	NN	O	O
syndrome	NN	O	O
.	NN	O	O
.	NN	O	O

Sialophorin	NN	O	O
,	NN	O	O
a	NN	O	O
surface	NN	O	O
sialoglycoprotein	NN	O	O
defective	NN	O	O
in	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
.	NN	O	O

The	NN	O	O
mAb	NN	O	O
L10	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
distribution	NN	O	O
and	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
sialophorin	NN	O	O
,	NN	O	O
the	NN	O	O
heavily	NN	O	O
glycosylated	NN	O	O
surface	NN	O	O
molecule	NN	O	O
that	NN	O	O
is	NN	O	O
deficient	NN	O	O
/	NN	O	O
defective	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
immunodeficiency	NN	O	I-Disease
Wiskott	NN	O	I-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Dual	NN	O	O
-	NN	O	O
parameter	NN	O	O
FACS	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
that	NN	O	O
sialophorin	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
CD4	NN	O	O
+	NN	O	O
and	NN	O	O
CD8	NN	O	O
+	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
on	NN	O	O
a	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
B	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
on	NN	O	O
all	NN	O	O
thymocytes	NN	O	O
,	NN	O	O
and	NN	O	O
on	NN	O	O
a	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
bone	NN	O	O
marrow	NN	O	O
cells	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
L10	NN	O	O
mAb	NN	O	O
stimulates	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
.	NN	O	O

The	NN	O	O
time	NN	O	O
course	NN	O	O
and	NN	O	O
magnitude	NN	O	O
of	NN	O	O
increased	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
by	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
L10	NN	O	O
mAb	NN	O	O
paralleled	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
anti	NN	O	O
-	NN	O	O
CD3	NN	O	O
mAb	NN	O	O
.	NN	O	O

Effective	NN	O	O
stimulation	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
monocytes	NN	O	O
and	NN	O	O
the	NN	O	O
Fc	NN	O	O
portion	NN	O	O
of	NN	O	O
L10	NN	O	O
mAb	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
by	NN	O	O
L10	NN	O	O
,	NN	O	O
like	NN	O	O
stimulation	NN	O	O
by	NN	O	O
anti	NN	O	O
-	NN	O	O
CD3	NN	O	O
mAb	NN	O	O
,	NN	O	O
involves	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
4F2	NN	O	O
,	NN	O	O
HLA	NN	O	O
-	NN	O	O
DR	NN	O	O
,	NN	O	O
and	NN	O	O
IL	NN	O	O
-	NN	O	O
2	NN	O	O
-	NN	O	O
R	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
sialophorin	NN	O	O
functions	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O
.	NN	O	O

Hypopigmentation	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Cutaneous	NN	O	B-Disease
and	NN	O	I-Disease
ocular	NN	O	I-Disease
pigmentation	NN	O	I-Disease
were	NN	O	O
evaluated	NN	O	O
in	NN	O	O
29	NN	O	O
individuals	NN	O	O
with	NN	O	O
the	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Criteria	NN	O	O
for	NN	O	O
hypopigmentation	NN	O	B-Disease
included	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
type	NN	O	O
I	NN	O	O
or	NN	O	O
II	NN	O	O
skin	NN	O	O
,	NN	O	O
the	NN	O	O
lightest	NN	O	O
skin	NN	O	O
type	NN	O	O
in	NN	O	O
the	NN	O	O
family	NN	O	O
by	NN	O	O
history	NN	O	O
,	NN	O	O
and	NN	O	O
iris	NN	O	O
translucency	NN	O	O
on	NN	O	O
globe	NN	O	O
transillumination	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
these	NN	O	O
criteria	NN	O	O
,	NN	O	O
48	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
individuals	NN	O	O
were	NN	O	O
hypopigmented	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
hypopigmentation	NN	O	B-Disease
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
small	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
on	NN	O	O
the	NN	O	O
proximal	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
this	NN	O	O
deletion	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
individuals	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
meet	NN	O	O
the	NN	O	O
full	NN	O	O
criteria	NN	O	O
for	NN	O	O
hypopigmentation	NN	O	B-Disease
.	NN	O	O

Hairbulb	NN	O	O
tyrosinase	NN	O	O
activity	NN	O	O
and	NN	O	O
glutathione	NN	O	O
content	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
urine	NN	O	O
cysteinyldopa	NN	O	O
excretion	NN	O	O
,	NN	O	O
were	NN	O	O
low	NN	O	O
in	NN	O	O
PWS	NN	O	B-Disease
individuals	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
hypopigmentation	NN	O	B-Disease
and	NN	O	O
did	NN	O	O
not	NN	O	O
separate	NN	O	O
these	NN	O	O
two	NN	O	O
groups	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
hypopigmentation	NN	O	B-Disease
is	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
proportion	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
PWS	NN	O	B-Disease
and	NN	O	O
that	NN	O	O
the	NN	O	O
hypopigmentation	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
hypopigmentation	NN	O	B-Disease
is	NN	O	O
unknown	NN	O	O
.	NN	O	O
.	NN	O	O

Phenotype	NN	O	O
heterogeneity	NN	O	O
among	NN	O	O
hemizygotes	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
biochemically	NN	O	O
screened	NN	O	O
for	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
two	NN	O	O
clinically	NN	O	O
,	NN	O	O
neurologically	NN	O	O
normal	NN	O	O
relatives	NN	O	O
of	NN	O	O
a	NN	O	O
boy	NN	O	O
affected	NN	O	O
by	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
;	NN	O	O
they	NN	O	O
were	NN	O	O
found	NN	O	O
repeatedly	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
biochemical	NN	O	O
defect	NN	O	O
of	NN	O	O
an	NN	O	O
ALD	NN	O	B-Disease
hemizygote	NN	O	O
.	NN	O	O

The	NN	O	O
assay	NN	O	O
consisted	NN	O	O
in	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
in	NN	O	O
lyophilized	NN	O	O
and	NN	O	O
reconstituted	NN	O	O
plasma	NN	O	O
.	NN	O	O

While	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
neurologic	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
leukodystrophy	NN	O	B-Disease
or	NN	O	O
myeloneuropathy	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
these	NN	O	O
hemizygotes	NN	O	O
,	NN	O	O
adrenocortical	NN	O	B-Disease
insufficiency	NN	O	I-Disease
provoking	NN	O	O
compensatory	NN	O	O
high	NN	O	O
ACTH	NN	O	O
release	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
both	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
should	NN	O	O
be	NN	O	O
taken	NN	O	O
into	NN	O	O
consideration	NN	O	O
when	NN	O	O
counseling	NN	O	O
families	NN	O	O
in	NN	O	O
which	NN	O	O
cases	NN	O	O
with	NN	O	O
clinically	NN	O	O
expressed	NN	O	O
ALD	NN	O	B-Disease
are	NN	O	O
represented	NN	O	O
in	NN	O	O
several	NN	O	O
generations	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
mapping	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Duchenne	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
to	NN	O	O
the	NN	O	O
mouse	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

The	NN	O	O
recent	NN	O	O
discovery	NN	O	O
of	NN	O	O
sequences	NN	O	O
at	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
in	NN	O	O
humans	NN	O	O
has	NN	O	O
opened	NN	O	O
up	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
detailed	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
in	NN	O	O
humans	NN	O	O
and	NN	O	O
in	NN	O	O
related	NN	O	O
mammalian	NN	O	O
species	NN	O	O
.	NN	O	O

Until	NN	O	O
relatively	NN	O	O
recently	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
obvious	NN	O	O
mouse	NN	O	O
model	NN	O	O
of	NN	O	O
this	NN	O	O
genetic	NN	O	B-Disease
disease	NN	O	I-Disease
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
mouse	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
mutant	NN	O	O
showing	NN	O	O
muscular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
mdx	NN	O	O
,	NN	O	O
has	NN	O	O
provided	NN	O	O
a	NN	O	O
candidate	NN	O	O
mouse	NN	O	O
genetic	NN	O	O
homologue	NN	O	O
to	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
;	NN	O	O
the	NN	O	O
relatively	NN	O	O
mild	NN	O	O
pathological	NN	O	O
features	NN	O	O
of	NN	O	O
mdx	NN	O	O
suggest	NN	O	O
it	NN	O	O
may	NN	O	O
have	NN	O	O
more	NN	O	O
in	NN	O	O
common	NN	O	O
with	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
type	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
human	NN	O	O
locus	NN	O	O
,	NN	O	O
however	NN	O	O
.	NN	O	O

But	NN	O	O
the	NN	O	O
close	NN	O	O
genetic	NN	O	O
linkage	NN	O	O
of	NN	O	O
mdx	NN	O	O
to	NN	O	O
G6PD	NN	O	O
and	NN	O	O
Hprt	NN	O	O
on	NN	O	O
the	NN	O	O
mouse	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
its	NN	O	O
comparatively	NN	O	O
mild	NN	O	O
pathology	NN	O	O
,	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
mdx	NN	O	O
mutation	NN	O	O
may	NN	O	O
instead	NN	O	O
correspond	NN	O	O
to	NN	O	O
Emery	NN	O	B-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
which	NN	O	O
itself	NN	O	O
is	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
DNA	NN	O	O
markers	NN	O	O
at	NN	O	O
Xq28	NN	O	O
-	NN	O	O
qter	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
G6PD	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Using	NN	O	O
an	NN	O	O
interspecific	NN	O	O
mouse	NN	O	O
domesticus	NN	O	O
/	NN	O	O
spretus	NN	O	O
cross	NN	O	O
,	NN	O	O
segregating	NN	O	O
for	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
markers	NN	O	O
on	NN	O	O
the	NN	O	O
mouse	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
positioned	NN	O	O
on	NN	O	O
the	NN	O	O
mouse	NN	O	O
X	NN	O	O
chromosome	NN	O	O
sequences	NN	O	O
homologous	NN	O	O
to	NN	O	O
a	NN	O	O
DMD	NN	O	B-Disease
cDNA	NN	O	O
clone	NN	O	O
.	NN	O	O

These	NN	O	O
sequences	NN	O	O
map	NN	O	O
provocatively	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
mdx	NN	O	O
mutation	NN	O	O
and	NN	O	O
unexpectedly	NN	O	O
distant	NN	O	O
from	NN	O	O
sparse	NN	O	O
fur	NN	O	O
,	NN	O	O
spf	NN	O	O
,	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
OTC	NN	O	O
(	NN	O	O
ornithine	NN	O	O
transcarbamylase	NN	O	O
)	NN	O	O
which	NN	O	O
is	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
DMD	NN	O	B-Disease
on	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O
.	NN	O	O

Localization	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
locus	NN	O	O
on	NN	O	O
the	NN	O	O
mouse	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

Recent	NN	O	O
progress	NN	O	O
has	NN	O	O
resulted	NN	O	O
in	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
mutated	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
and	NN	O	I-Disease
the	NN	O	I-Disease
milder	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophies	NN	O	I-Disease
being	NN	O	O
cloned	NN	O	O
and	NN	O	O
has	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
itself	NN	O	O
extends	NN	O	O
over	NN	O	O
1	NN	O	O
,	NN	O	O
000	NN	O	O
to	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
kilobases	NN	O	O
(	NN	O	O
kb	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
how	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
affect	NN	O	O
muscle	NN	O	O
development	NN	O	O
and	NN	O	O
integrity	NN	O	O
,	NN	O	O
it	NN	O	O
would	NN	O	O
be	NN	O	O
of	NN	O	O
interest	NN	O	O
to	NN	O	O
have	NN	O	O
available	NN	O	O
a	NN	O	O
mouse	NN	O	O
model	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
mouse	NN	O	O
mdx	NN	O	O
mutation	NN	O	O
affects	NN	O	O
muscle	NN	O	O
and	NN	O	O
confers	NN	O	O
a	NN	O	O
mild	NN	O	O
dystrophic	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
whether	NN	O	O
this	NN	O	O
mutation	NN	O	O
is	NN	O	O
equivalent	NN	O	O
to	NN	O	O
Duchenne	NN	O	B-Disease
/	NN	O	I-Disease
Becker	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
in	NN	O	O
man	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
two	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
that	NN	O	O
cross	NN	O	O
-	NN	O	O
hybridize	NN	O	O
to	NN	O	O
mouse	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
sequences	NN	O	O
to	NN	O	O
localize	NN	O	O
the	NN	O	O
gene	NN	O	O
homologous	NN	O	O
to	NN	O	O
DMD	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
.	NN	O	O

Both	NN	O	O
sequences	NN	O	O
map	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
10	NN	O	O
centimorgan	NN	O	O
lying	NN	O	O
between	NN	O	O
the	NN	O	O
Tabby	NN	O	O
(	NN	O	O
Ta	NN	O	O
)	NN	O	O
and	NN	O	O
St14	NN	O	O
-	NN	O	O
1	NN	O	O
(	NN	O	O
DxPas8	NN	O	O
)	NN	O	O
loci	NN	O	O
,	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
phosphorylase	NN	O	O
b	NN	O	O
kinase	NN	O	O
locus	NN	O	O
(	NN	O	O
Phk	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
analogy	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
around	NN	O	O
the	NN	O	O
G6pd	NN	O	O
and	NN	O	O
St14	NN	O	O
-	NN	O	O
1	NN	O	O
loci	NN	O	O
may	NN	O	O
contain	NN	O	O
two	NN	O	O
genes	NN	O	O
corresponding	NN	O	O
to	NN	O	O
distinct	NN	O	O
human	NN	O	O
myopathies	NN	O	B-Disease
Emery	NN	O	B-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
St14	NN	O	O
-	NN	O	O
1	NN	O	O
in	NN	O	O
man	NN	O	O
and	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
homologue	NN	O	O
described	NN	O	O
here	NN	O	O
.	NN	O	O
.	NN	O	O

GT	NN	O	O
to	NN	O	O
AT	NN	O	O
transition	NN	O	O
at	NN	O	O
a	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
causes	NN	O	O
skipping	NN	O	O
of	NN	O	O
the	NN	O	O
preceding	NN	O	O
exon	NN	O	O
in	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O

Classical	NN	O	B-Disease
Phenylketonuria	NN	O	I-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
human	NN	O	I-Disease
genetic	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hepatic	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
isolated	NN	O	O
several	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
from	NN	O	O
a	NN	O	O
PKU	NN	O	B-Disease
carrier	NN	O	O
individual	NN	O	O
and	NN	O	O
showed	NN	O	O
that	NN	O	O
they	NN	O	O
contained	NN	O	O
an	NN	O	O
internal	NN	O	O
116	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
,	NN	O	O
corresponding	NN	O	O
precisely	NN	O	O
to	NN	O	O
exon	NN	O	O
12	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
chromosomal	NN	O	O
PAH	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
causes	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
protein	NN	O	O
lacking	NN	O	O
the	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
52	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

Gene	NN	O	O
transfer	NN	O	O
and	NN	O	O
expression	NN	O	O
studies	NN	O	O
using	NN	O	O
the	NN	O	O
mutant	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
deletion	NN	O	O
abolishes	NN	O	O
PAH	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
protein	NN	O	O
instability	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
,	NN	O	O
the	NN	O	O
mutant	NN	O	O
chromosomal	NN	O	O
PAH	NN	O	O
gene	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
this	NN	O	O
individual	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
a	NN	O	O
GT	NN	O	O
-	NN	O	O
-	NN	O	O
greater	NN	O	O
than	NN	O	O
AT	NN	O	O
substitution	NN	O	O
at	NN	O	O
the	NN	O	O
5	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
12	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
liver	NN	O	O
is	NN	O	O
the	NN	O	O
skipping	NN	O	O
of	NN	O	O
the	NN	O	O
preceding	NN	O	O
exon	NN	O	O
during	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
.	NN	O	O
.	NN	O	O

Conservation	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

A	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
transcript	NN	O	O
from	NN	O	O
human	NN	O	O
fetal	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
and	NN	O	O
mouse	NN	O	O
adult	NN	O	O
heart	NN	O	O
was	NN	O	O
sequenced	NN	O	O
,	NN	O	O
representing	NN	O	O
approximately	NN	O	O
25	NN	O	O
percent	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
,	NN	O	O
14	NN	O	O
-	NN	O	O
kb	NN	O	O
DMD	NN	O	B-Disease
transcript	NN	O	O
.	NN	O	O

The	NN	O	O
nucleic	NN	O	O
acid	NN	O	O
and	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
two	NN	O	O
species	NN	O	O
are	NN	O	O
nearly	NN	O	O
90	NN	O	O
percent	NN	O	O
homologous	NN	O	O
.	NN	O	O

The	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
that	NN	O	O
is	NN	O	O
predicted	NN	O	O
from	NN	O	O
this	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
product	NN	O	O
might	NN	O	O
serve	NN	O	O
a	NN	O	O
structural	NN	O	O
role	NN	O	O
in	NN	O	O
muscle	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
abundance	NN	O	O
and	NN	O	O
tissue	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
protein	NN	O	O
is	NN	O	O
not	NN	O	O
nebulin	NN	O	O
.	NN	O	O
.	NN	O	O

Familial	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
with	NN	O	O
apparently	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
4	NN	O	O
sibs	NN	O	O
(	NN	O	O
2F	NN	O	O
,	NN	O	O
2M	NN	O	O
)	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Diagnosis	NN	O	O
was	NN	O	O
made	NN	O	O
clinically	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
history	NN	O	O
,	NN	O	O
behavior	NN	O	O
,	NN	O	O
and	NN	O	O
physical	NN	O	O
findings	NN	O	O
in	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
sibs	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
child	NN	O	O
had	NN	O	O
died	NN	O	O
at	NN	O	O
age	NN	O	O
10	NN	O	O
months	NN	O	O
with	NN	O	O
a	NN	O	O
history	NN	O	O
and	NN	O	O
clinical	NN	O	O
findings	NN	O	O
typical	NN	O	O
of	NN	O	O
first	NN	O	O
phase	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

Results	NN	O	O
of	NN	O	O
chromosome	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
parents	NN	O	O
and	NN	O	O
surviving	NN	O	O
sibs	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
implications	NN	O	O
of	NN	O	O
this	NN	O	O
unusual	NN	O	O
familial	NN	O	O
occurrence	NN	O	O
for	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
are	NN	O	O
discussed	NN	O	O
.	NN	O	O
.	NN	O	O

Nebulin	NN	O	O
and	NN	O	O
titin	NN	O	O
expression	NN	O	O
in	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
appears	NN	O	O
normal	NN	O	O
.	NN	O	O

Monoclonal	NN	O	O
antibodies	NN	O	O
which	NN	O	O
recognize	NN	O	O
different	NN	O	O
epitopes	NN	O	O
on	NN	O	O
either	NN	O	O
titin	NN	O	O
or	NN	O	O
nebulin	NN	O	O
show	NN	O	O
normal	NN	O	O
staining	NN	O	O
patterns	NN	O	O
on	NN	O	O
frozen	NN	O	O
sections	NN	O	O
of	NN	O	O
three	NN	O	O
muscle	NN	O	O
biopsies	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Gel	NN	O	O
electrophoresis	NN	O	O
and	NN	O	O
immunoblotting	NN	O	O
performed	NN	O	O
on	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
muscle	NN	O	O
biopsies	NN	O	O
show	NN	O	O
the	NN	O	O
normal	NN	O	O
pattern	NN	O	O
of	NN	O	O
titin	NN	O	O
and	NN	O	O
nebulin	NN	O	O
polypeptides	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
donor	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
biopsies	NN	O	O
has	NN	O	O
a	NN	O	O
large	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
-	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
argue	NN	O	O
against	NN	O	O
the	NN	O	O
recent	NN	O	O
proposal	NN	O	O
that	NN	O	O
nebulin	NN	O	O
is	NN	O	O
the	NN	O	O
gene	NN	O	O
mutated	NN	O	O
in	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Heterozygous	NN	O	B-Disease
C2	NN	O	I-Disease
deficiency	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
angioedema	NN	O	B-Disease
,	NN	O	O
myasthenia	NN	O	B-Disease
gravis	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
systemic	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
myasthenia	NN	O	B-Disease
gravis	NN	O	I-Disease
,	NN	O	O
systemic	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
angioedema	NN	O	B-Disease
associated	NN	O	O
with	NN	O	O
heterozygous	NN	O	O
complement	NN	O	B-Disease
factor	NN	O	I-Disease
2	NN	O	I-Disease
(	NN	O	I-Disease
C2	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
association	NN	O	O
is	NN	O	O
controversial	NN	O	O
,	NN	O	O
though	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
with	NN	O	O
certain	NN	O	O
histocompatibility	NN	O	O
antigens	NN	O	O
suggests	NN	O	O
possible	NN	O	O
linkage	NN	O	O
to	NN	O	O
immune	NN	O	O
response	NN	O	O
genes	NN	O	O
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
heterozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
association	NN	O	O
with	NN	O	O
this	NN	O	O
combination	NN	O	O
of	NN	O	O
autoimmune	NN	O	B-Disease
disorders	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
we	NN	O	O
discuss	NN	O	O
the	NN	O	O
aetiological	NN	O	O
implications	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
an	NN	O	O
inherited	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
third	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
Brittany	NN	O	O
spaniel	NN	O	O
dogs	NN	O	O
.	NN	O	O

Genetically	NN	O	O
determined	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Brittany	NN	O	O
spaniel	NN	O	O
dogs	NN	O	O
shares	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
biochemical	NN	O	O
and	NN	O	O
clinical	NN	O	O
characteristics	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
disorder	NN	O	O
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
null	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
allelic	NN	O	O
to	NN	O	O
the	NN	O	O
structural	NN	O	O
gene	NN	O	O
for	NN	O	O
C3	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
locus	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
used	NN	O	O
allotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
canine	NN	O	O
C3	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
these	NN	O	O
dogs	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
null	NN	O	O
gene	NN	O	O
allelic	NN	O	O
to	NN	O	O
the	NN	O	O
structural	NN	O	O
gene	NN	O	O
for	NN	O	O
C3	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
preliminary	NN	O	O
pedigree	NN	O	O
analysis	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
canine	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
apparently	NN	O	O
not	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
complex	NN	O	O
of	NN	O	O
the	NN	O	O
dog	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Brittany	NN	O	O
spaniel	NN	O	O
dogs	NN	O	O
not	NN	O	O
only	NN	O	O
shares	NN	O	O
biochemical	NN	O	O
and	NN	O	O
clinical	NN	O	O
features	NN	O	O
with	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
humans	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
shares	NN	O	O
some	NN	O	O
genetic	NN	O	O
characteristics	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
disorder	NN	O	O
.	NN	O	O
.	NN	O	O

Gene	NN	O	O
transfer	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
genetic	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
enzyme	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
complementary	NN	O	O
DNA	NN	O	O
clone	NN	O	O
of	NN	O	O
human	NN	O	O
PAH	NN	O	O
was	NN	O	O
inserted	NN	O	O
into	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
expression	NN	O	O
vector	NN	O	O
and	NN	O	O
transferred	NN	O	O
into	NN	O	O
mouse	NN	O	O
NIH3T3	NN	O	O
cells	NN	O	O
which	NN	O	O
do	NN	O	O
not	NN	O	O
normally	NN	O	O
express	NN	O	O
PAH	NN	O	O
.	NN	O	O

The	NN	O	O
transformed	NN	O	O
mouse	NN	O	O
cells	NN	O	O
expressed	NN	O	O
PAH	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
,	NN	O	O
immunoreactive	NN	O	O
protein	NN	O	O
,	NN	O	O
and	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
that	NN	O	O
are	NN	O	O
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
human	NN	O	O
liver	NN	O	O
products	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
gene	NN	O	O
contains	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
necessary	NN	O	O
genetic	NN	O	O
information	NN	O	O
to	NN	O	O
code	NN	O	O
for	NN	O	O
functional	NN	O	O
PAH	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
PAH	NN	O	O
probe	NN	O	O
in	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
detection	NN	O	O
of	NN	O	O
carriers	NN	O	O
,	NN	O	O
to	NN	O	O
provide	NN	O	O
new	NN	O	O
opportunities	NN	O	O
for	NN	O	O
the	NN	O	O
biochemical	NN	O	O
characterization	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
mutant	NN	O	O
enzymes	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
investigation	NN	O	O
of	NN	O	O
alternative	NN	O	O
genetic	NN	O	O
therapies	NN	O	O
for	NN	O	O
PKU	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Regional	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
phenylketonuria	NN	O	B-Disease
locus	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
genome	NN	O	O
.	NN	O	O

Phenylketonuria	NN	O	B-Disease
(	NN	O	O
PKU	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	O
amino	NN	O	O
acid	NN	O	O
metabolism	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
hepatic	NN	O	I-Disease
enzyme	NN	O	I-Disease
phenylalanine	NN	O	I-Disease
hydroxylase	NN	O	I-Disease
(	NN	O	O
PAH	NN	O	O
;	NN	O	O
phenylalanine	NN	O	O
4	NN	O	O
-	NN	O	O
monooxygenase	NN	O	O
,	NN	O	O
EC	NN	O	O
1	NN	O	O
.	NN	O	O
14	NN	O	O
.	NN	O	O
16	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
cDNA	NN	O	O
clone	NN	O	O
for	NN	O	O
human	NN	O	O
PAH	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
assign	NN	O	O
the	NN	O	O
corresponding	NN	O	O
gene	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
regional	NN	O	O
map	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
locus	NN	O	O
and	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
on	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O
,	NN	O	O
DNA	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
human	NN	O	O
-	NN	O	O
hamster	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
with	NN	O	O
various	NN	O	O
deletions	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O
and	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
using	NN	O	O
the	NN	O	O
human	NN	O	O
cDNA	NN	O	O
PAH	NN	O	O
clone	NN	O	O
as	NN	O	O
a	NN	O	O
hybridization	NN	O	O
probe	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
detailed	NN	O	O
biochemical	NN	O	O
and	NN	O	O
cytogenetic	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
hybrid	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
12	NN	O	O
containing	NN	O	O
the	NN	O	O
human	NN	O	O
PAH	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
defined	NN	O	O
as	NN	O	O
12q14	NN	O	O
.	NN	O	O

3	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
qter	NN	O	O
3	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O

The	NN	O	O
PAH	NN	O	O
map	NN	O	O
position	NN	O	O
on	NN	O	O
chromosome	NN	O	O
12	NN	O	O
was	NN	O	O
further	NN	O	O
localized	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
of	NN	O	O
125I	NN	O	O
-	NN	O	O
labeled	NN	O	O
human	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
to	NN	O	O
chromosomes	NN	O	O
prepared	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

Results	NN	O	O
of	NN	O	O
these	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
12	NN	O	O
containing	NN	O	O
the	NN	O	O
PAH	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
PKU	NN	O	B-Disease
locus	NN	O	O
in	NN	O	O
man	NN	O	O
is	NN	O	O
12q22	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
-	NN	O	O
12q24	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
not	NN	O	O
only	NN	O	O
provide	NN	O	O
a	NN	O	O
regionalized	NN	O	O
map	NN	O	O
position	NN	O	O
for	NN	O	O
a	NN	O	O
major	NN	O	O
human	NN	O	O
disease	NN	O	O
locus	NN	O	O
but	NN	O	O
also	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
reference	NN	O	O
point	NN	O	O
for	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
other	NN	O	O
DNA	NN	O	O
markers	NN	O	O
on	NN	O	O
human	NN	O	O
chromosome	NN	O	O
12	NN	O	O

Heterogeneity	NN	O	O
of	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
von	NN	O	I-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
subgroup	NN	O	O
with	NN	O	O
an	NN	O	O
abnormal	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
.	NN	O	O

Type	NN	O	B-Disease
I	NN	O	I-Disease
von	NN	O	I-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
vWD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
equally	NN	O	O
low	NN	O	O
plasma	NN	O	O
concentrations	NN	O	O
of	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
antigen	NN	O	O
(	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
)	NN	O	O
and	NN	O	O
ristocetin	NN	O	O
cofactor	NN	O	O
(	NN	O	O
RiCof	NN	O	O
)	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
all	NN	O	O
vWF	NN	O	O
multimers	NN	O	O
in	NN	O	O
sodium	NN	O	O
dodecyl	NN	O	O
sulfate	NN	O	O
(	NN	O	O
SDS	NN	O	O
)	NN	O	O
-	NN	O	O
agarose	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
.	NN	O	O

For	NN	O	O
17	NN	O	O
patients	NN	O	O
(	NN	O	O
13	NN	O	O
kindreds	NN	O	O
)	NN	O	O
diagnosed	NN	O	O
with	NN	O	O
these	NN	O	O
criteria	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
platelet	NN	O	O
contents	NN	O	O
of	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
and	NN	O	O
RiCof	NN	O	O
and	NN	O	O
the	NN	O	O
changes	NN	O	O
of	NN	O	O
these	NN	O	O
in	NN	O	O
plasma	NN	O	O
after	NN	O	O
DDAVP	NN	O	O
infusion	NN	O	O
.	NN	O	O

Platelet	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
and	NN	O	O
RiCof	NN	O	O
were	NN	O	O
normal	NN	O	O
in	NN	O	O
four	NN	O	O
kindreds	NN	O	O
(	NN	O	O
called	NN	O	O
"	NN	O	O
platelet	NN	O	O
normal	NN	O	O
"	NN	O	O
subgroup	NN	O	O
)	NN	O	O
;	NN	O	O
following	NN	O	O
1	NN	O	O
-	NN	O	O
deamino	NN	O	O
-	NN	O	O
8	NN	O	O
-	NN	O	O
D	NN	O	O
-	NN	O	O
arginine	NN	O	O
vasopressin	NN	O	O
;	NN	O	O
plasma	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
,	NN	O	O
RiCof	NN	O	O
and	NN	O	O
the	NN	O	O
bleeding	NN	O	O
time	NN	O	O
(	NN	O	O
BT	NN	O	O
)	NN	O	O
became	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
six	NN	O	O
kindreds	NN	O	O
,	NN	O	O
platelet	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
and	NN	O	O
RiCof	NN	O	O
were	NN	O	O
equally	NN	O	O
low	NN	O	O
(	NN	O	O
platelet	NN	O	O
low	NN	O	O
)	NN	O	O
;	NN	O	O
after	NN	O	O
DDAVP	NN	O	O
,	NN	O	O
plasma	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
and	NN	O	O
RiCof	NN	O	O
remained	NN	O	O
low	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
BT	NN	O	O
was	NN	O	O
prolonged	NN	O	O
.	NN	O	O

In	NN	O	O
three	NN	O	O
additional	NN	O	O
kindreds	NN	O	O
,	NN	O	O
platelets	NN	O	O
contained	NN	O	O
normal	NN	O	O
concentrations	NN	O	O
of	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
,	NN	O	O
but	NN	O	O
RiCof	NN	O	O
was	NN	O	O
very	NN	O	O
low	NN	O	O
(	NN	O	O
platelet	NN	O	O
discordant	NN	O	O
)	NN	O	O
;	NN	O	O
even	NN	O	O
though	NN	O	O
a	NN	O	O
complete	NN	O	O
set	NN	O	O
of	NN	O	O
multimers	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
plasma	NN	O	O
and	NN	O	O
platelets	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
relatively	NN	O	O
small	NN	O	O
amount	NN	O	O
of	NN	O	O
large	NN	O	O
multimers	NN	O	O
.	NN	O	O

After	NN	O	O
DDAVP	NN	O	O
,	NN	O	O
plasma	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
became	NN	O	O
normal	NN	O	O
,	NN	O	O
but	NN	O	O
RiCof	NN	O	O
remained	NN	O	O
low	NN	O	O
and	NN	O	O
the	NN	O	O
BT	NN	O	O
was	NN	O	O
very	NN	O	O
prolonged	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
there	NN	O	O
can	NN	O	O
be	NN	O	O
an	NN	O	O
abnormal	NN	O	O
vWF	NN	O	O
(	NN	O	O
RiCof	NN	O	O
less	NN	O	O
than	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
)	NN	O	O
even	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
vWD	NN	O	I-Disease
,	NN	O	O
coexisting	NN	O	O
with	NN	O	O
a	NN	O	O
complete	NN	O	O
set	NN	O	O
of	NN	O	O
vWF	NN	O	O
multimers	NN	O	O
(	NN	O	O
platelet	NN	O	O
discordant	NN	O	O
)	NN	O	O
;	NN	O	O
that	NN	O	O
the	NN	O	O
abnormal	NN	O	O
vWF	NN	O	O
can	NN	O	O
be	NN	O	O
shown	NN	O	O
more	NN	O	O
clearly	NN	O	O
in	NN	O	O
platelets	NN	O	O
than	NN	O	O
in	NN	O	O
plasma	NN	O	O
or	NN	O	O
else	NN	O	O
in	NN	O	O
plasma	NN	O	O
after	NN	O	O
DDAVP	NN	O	O
infusion	NN	O	O
;	NN	O	O
and	NN	O	O
that	NN	O	O
DDAVP	NN	O	O
normalizes	NN	O	O
the	NN	O	O
BT	NN	O	O
only	NN	O	O
in	NN	O	O
those	NN	O	O
patients	NN	O	O
with	NN	O	O
normal	NN	O	O
platelet	NN	O	O
levels	NN	O	O
of	NN	O	O
both	NN	O	O
vWF	NN	O	O
Ag	NN	O	O
and	NN	O	O
RiCof	NN	O	O
(	NN	O	O
platelet	NN	O	O
normal	NN	O	O
)	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
brought	NN	O	O
together	NN	O	O
information	NN	O	O
on	NN	O	O
864	NN	O	O
affected	NN	O	O
individuals	NN	O	O
in	NN	O	O
164	NN	O	O
families	NN	O	O
(	NN	O	O
including	NN	O	O
three	NN	O	O
new	NN	O	O
pedigrees	NN	O	O
)	NN	O	O
reported	NN	O	O
in	NN	O	O
the	NN	O	O
137	NN	O	O
year	NN	O	O
period	NN	O	O
since	NN	O	O
1845	NN	O	O
when	NN	O	O
Demarquay	NN	O	O
first	NN	O	O
described	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
what	NN	O	O
was	NN	O	O
later	NN	O	O
called	NN	O	O
van	NN	O	B-Disease
der	NN	O	I-Disease
Woude	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
VWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Both	NN	O	O
types	NN	O	O
of	NN	O	O
oral	NN	O	B-Disease
cleft	NN	O	I-Disease
,	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
(	NN	O	O
CP	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
cleft	NN	O	B-Disease
lip	NN	O	I-Disease
with	NN	O	O
or	NN	O	O
without	NN	O	O
CP	NN	O	B-Disease
(	NN	O	O
CLP	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
segregate	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
together	NN	O	O
with	NN	O	O
lower	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
or	NN	O	O
fistulae	NN	O	B-Disease
in	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
mode	NN	O	O
with	NN	O	O
high	NN	O	O
penetrance	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
K	NN	O	O
=	NN	O	O
.	NN	O	O

89	NN	O	O
and	NN	O	O
.	NN	O	O

99	NN	O	O
by	NN	O	O
different	NN	O	O
methods	NN	O	O
.	NN	O	O

Cleft	NN	O	O
types	NN	O	O
(	NN	O	O
CLP	NN	O	B-Disease
and	NN	O	O
CP	NN	O	B-Disease
)	NN	O	O
occur	NN	O	O
in	NN	O	O
VWS	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
same	NN	O	O
proportions	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
non	NN	O	O
-	NN	O	O
VWS	NN	O	O
population	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
about	NN	O	O
twice	NN	O	O
as	NN	O	O
many	NN	O	O
cleft	NN	O	O
-	NN	O	O
bearing	NN	O	O
individuals	NN	O	O
have	NN	O	O
CLP	NN	O	B-Disease
as	NN	O	O
have	NN	O	O
CP	NN	O	B-Disease
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
we	NN	O	O
do	NN	O	O
not	NN	O	O
find	NN	O	O
the	NN	O	O
usually	NN	O	O
observed	NN	O	O
excess	NN	O	O
of	NN	O	O
females	NN	O	O
with	NN	O	O
CP	NN	O	B-Disease
and	NN	O	O
excess	NN	O	O
of	NN	O	O
males	NN	O	O
with	NN	O	O
CLP	NN	O	B-Disease
;	NN	O	O
in	NN	O	O
VWS	NN	O	B-Disease
the	NN	O	O
sex	NN	O	O
ratios	NN	O	O
are	NN	O	O
more	NN	O	O
nearly	NN	O	O
equal	NN	O	O
.	NN	O	O

Lip	NN	O	B-Disease
pits	NN	O	I-Disease
also	NN	O	O
are	NN	O	O
equally	NN	O	O
distributed	NN	O	O
between	NN	O	O
the	NN	O	O
sexes	NN	O	O
.	NN	O	O

Affected	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O
are	NN	O	O
equally	NN	O	O
likely	NN	O	O
to	NN	O	O
transmit	NN	O	O
VWS	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
less	NN	O	O
severely	NN	O	O
affected	NN	O	O
individuals	NN	O	O
among	NN	O	O
transmitters	NN	O	O
and	NN	O	O
a	NN	O	O
deficiency	NN	O	O
of	NN	O	O
more	NN	O	O
severely	NN	O	O
affected	NN	O	O
,	NN	O	O
brought	NN	O	O
about	NN	O	O
by	NN	O	O
a	NN	O	O
proband	NN	O	O
bias	NN	O	O
and	NN	O	O
differential	NN	O	O
fecundity	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
VWS	NN	O	B-Disease
is	NN	O	O
significantly	NN	O	O
modified	NN	O	O
by	NN	O	O
the	NN	O	O
genetic	NN	O	O
background	NN	O	O
More	NN	O	O
extreme	NN	O	O
phenotypes	NN	O	O
in	NN	O	O
parents	NN	O	O
tend	NN	O	O
to	NN	O	O
produce	NN	O	O
more	NN	O	O
extreme	NN	O	O
expression	NN	O	O
in	NN	O	O
their	NN	O	O
children	NN	O	O
.	NN	O	O

For	NN	O	O
a	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
carrier	NN	O	O
the	NN	O	O
relative	NN	O	O
risk	NN	O	O
of	NN	O	O
transmitting	NN	O	O
a	NN	O	O
cleft	NN	O	O
is	NN	O	O
26	NN	O	O
.	NN	O	O

45	NN	O	O
%	NN	O	O
;	NN	O	O
that	NN	O	O
of	NN	O	O
transmitting	NN	O	O
lower	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
is	NN	O	O
23	NN	O	O
.	NN	O	O

55	NN	O	O
%	NN	O	O
.	NN	O	O

Three	NN	O	O
pedigrees	NN	O	O
of	NN	O	O
lip	NN	O	B-Disease
pits	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
literature	NN	O	O
show	NN	O	O
no	NN	O	O
clefts	NN	O	O
among	NN	O	O
a	NN	O	O
significant	NN	O	O
number	NN	O	O
of	NN	O	O
affected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Control	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
VWS	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
three	NN	O	O
target	NN	O	O
tissues	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
independent	NN	O	O
and	NN	O	O
separately	NN	O	O
designated	NN	O	O
.	NN	O	O

Mutation	NN	O	O
rate	NN	O	O
of	NN	O	O
the	NN	O	O
VWS	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
calculated	NN	O	O
to	NN	O	O
be	NN	O	O
1	NN	O	O
.	NN	O	O

8	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-	NN	O	O
5	NN	O	O
)	NN	O	O

Hereditary	NN	O	B-Disease
C7	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Diagnosis	NN	O	O
and	NN	O	O
HLA	NN	O	O
studies	NN	O	O
in	NN	O	O
a	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
family	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	O
of	NN	O	O
a	NN	O	O
44	NN	O	O
-	NN	O	O
yr	NN	O	O
-	NN	O	O
old	NN	O	O
woman	NN	O	O
of	NN	O	O
French	NN	O	O
-	NN	O	O
Canadian	NN	O	O
descent	NN	O	O
having	NN	O	O
a	NN	O	O
B	NN	O	O
-	NN	O	O
27	NN	O	O
positive	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
was	NN	O	O
deficient	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C7	NN	O	O
)	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
hemolytic	NN	O	O
and	NN	O	O
immunochemical	NN	O	O
methods	NN	O	O
.	NN	O	O

No	NN	O	O
inhibitor	NN	O	O
against	NN	O	O
C7	NN	O	O
was	NN	O	O
detected	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
all	NN	O	O
other	NN	O	O
complement	NN	O	O
components	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

No	NN	O	O
deficiency	NN	O	O
in	NN	O	O
the	NN	O	O
opsonic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
serum	NN	O	O
was	NN	O	O
found	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
basic	NN	O	O
coagulation	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

On	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
family	NN	O	O
,	NN	O	O
two	NN	O	O
sisters	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
deficiency	NN	O	O
but	NN	O	O
were	NN	O	O
otherwise	NN	O	O
in	NN	O	O
good	NN	O	O
health	NN	O	O
.	NN	O	O

The	NN	O	O
seven	NN	O	O
other	NN	O	O
siblings	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
while	NN	O	O
the	NN	O	O
paternal	NN	O	O
aunt	NN	O	O
had	NN	O	O
a	NN	O	O
normal	NN	O	O
C7	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
third	NN	O	O
generation	NN	O	O
,	NN	O	O
six	NN	O	O
children	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
homozygous	NN	O	O
sisters	NN	O	O
and	NN	O	O
five	NN	O	O
children	NN	O	O
of	NN	O	O
heterozygotes	NN	O	O
were	NN	O	O
available	NN	O	O
for	NN	O	O
testing	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
HLA	NN	O	O
antigens	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
22	NN	O	O
subjects	NN	O	O
and	NN	O	O
in	NN	O	O
three	NN	O	O
spouses	NN	O	O
indicated	NN	O	O
no	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
CM	NN	O	B-Disease
deficienty	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
HLA	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
occurrence	NN	O	O
of	NN	O	O
two	NN	O	O
hereditary	NN	O	O
complement	NN	O	B-Disease
deficiencies	NN	O	I-Disease
(	NN	O	I-Disease
C2	NN	O	I-Disease
and	NN	O	I-Disease
C7	NN	O	I-Disease
)	NN	O	I-Disease
was	NN	O	O
discovered	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
of	NN	O	O
this	NN	O	O
remarkable	NN	O	O
kindred	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
defect	NN	O	B-Disease
in	NN	O	I-Disease
secretion	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
C5	NN	O	I-Disease
in	NN	O	O
mice	NN	O	O
.	NN	O	O

A	NN	O	O
genetic	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fifth	NN	O	I-Disease
(	NN	O	I-Disease
C5	NN	O	I-Disease
)	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement1	NN	O	I-Disease
-	NN	O	I-Disease
3	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
serum	NN	O	O
glycoprotein	NN	O	O
of	NN	O	O
molecular	NN	O	O
weight	NN	O	O
(	NN	O	O
MW	NN	O	O
)	NN	O	O
220	NN	O	O
,	NN	O	O
000	NN	O	O
(	NN	O	O
ref	NN	O	O
.	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
39	NN	O	O
%	NN	O	O
of	NN	O	O
inbred	NN	O	O
strains	NN	O	O
of	NN	O	O
mice3	NN	O	O
.	NN	O	O

Sera	NN	O	O
of	NN	O	O
deficient	NN	O	O
mice	NN	O	O
lack	NN	O	O
detectable	NN	O	O
C5	NN	O	O
activity	NN	O	O
and	NN	O	O
protein2	NN	O	O
,	NN	O	O
3	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
deficient	NN	O	O
mice	NN	O	O
produce	NN	O	O
antibody	NN	O	O
to	NN	O	O
mouse	NN	O	O
C5	NN	O	O
when	NN	O	O
injected	NN	O	O
with	NN	O	O
sera	NN	O	O
from	NN	O	O
C5	NN	O	O
sufficient	NN	O	O
(	NN	O	O
normal	NN	O	O
)	NN	O	O
strains	NN	O	O
.	NN	O	O

Levy	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O

5	NN	O	O
showed	NN	O	O
that	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
between	NN	O	O
C5	NN	O	O
deficient	NN	O	O
(	NN	O	O
B10	NN	O	O
.	NN	O	O
D2	NN	O	O
/	NN	O	O
old	NN	O	O
line	NN	O	O
)	NN	O	O
macrophages	NN	O	O
and	NN	O	O
either	NN	O	O
C5	NN	O	O
sufficient	NN	O	O
(	NN	O	O
B10	NN	O	O
.	NN	O	O
D2	NN	O	O
/	NN	O	O
new	NN	O	O
line	NN	O	O
)	NN	O	O
mouse	NN	O	O
kidney	NN	O	O
or	NN	O	O
chicken	NN	O	O
erythroblasts	NN	O	O
secreted	NN	O	O
haemolytically	NN	O	O
active	NN	O	O
mouse	NN	O	O
C5	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Several	NN	O	O
possible	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
findings	NN	O	O
were	NN	O	O
considered	NN	O	O
,	NN	O	O
but	NN	O	O
insufficient	NN	O	O
direct	NN	O	O
data	NN	O	O
were	NN	O	O
available	NN	O	O
to	NN	O	O
choose	NN	O	O
among	NN	O	O
them	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
mouse	NN	O	O
(	NN	O	O
CD	NN	O	O
.	NN	O	O
1	NN	O	O
strain	NN	O	O
)	NN	O	O
peritoneal	NN	O	O
cells	NN	O	O
in	NN	O	O
culture	NN	O	O
synthesise	NN	O	O
and	NN	O	O
secrete	NN	O	O
a	NN	O	O
single	NN	O	O
chain	NN	O	O
precursor	NN	O	O
,	NN	O	O
pro	NN	O	O
-	NN	O	O
C5	NN	O	O
(	NN	O	O
MW	NN	O	O
approximately	NN	O	O
210	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
-	NN	O	O
chain	NN	O	O
(	NN	O	O
alpha	NN	O	O
chain	NN	O	O
,	NN	O	O
125	NN	O	O
,	NN	O	O
000	NN	O	O
and	NN	O	O
beta	NN	O	O
chain	NN	O	O
83	NN	O	O
,	NN	O	O
000	NN	O	O
MW	NN	O	O
)	NN	O	O
C5	NN	O	O
protein6	NN	O	O
.	NN	O	O

Radiolabelled	NN	O	O
precursor	NN	O	O
C5	NN	O	O
was	NN	O	O
contained	NN	O	O
within	NN	O	O
the	NN	O	O
cells	NN	O	O
and	NN	O	O
was	NN	O	O
secreted	NN	O	O
into	NN	O	O
the	NN	O	O
tissue	NN	O	O
culture	NN	O	O
media	NN	O	O
.	NN	O	O

Using	NN	O	O
similar	NN	O	O
methods	NN	O	O
,	NN	O	O
we	NN	O	O
now	NN	O	O
find	NN	O	O
that	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
each	NN	O	O
of	NN	O	O
five	NN	O	O
different	NN	O	O
mouse	NN	O	O
strains	NN	O	O
(	NN	O	O
AKR	NN	O	O
,	NN	O	O
SWR	NN	O	O
,	NN	O	O
DBA	NN	O	O
/	NN	O	O
2J8	NN	O	O
A	NN	O	O
/	NN	O	O
HeJ	NN	O	O
and	NN	O	O
B10	NN	O	O
.	NN	O	O
D2	NN	O	O
/	NN	O	O
old	NN	O	O
line	NN	O	O
)	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
failure	NN	O	O
in	NN	O	O
secretion	NN	O	O
of	NN	O	O
C5	NN	O	O
protein	NN	O	O
and	NN	O	O
not	NN	O	O
to	NN	O	O
a	NN	O	O
failure	NN	O	O
in	NN	O	O
biosynthesis	NN	O	O
of	NN	O	O
pro	NN	O	O
-	NN	O	O
C5	NN	O	O

Recurrent	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
with	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
sixth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
:	NN	O	O
an	NN	O	O
evaluation	NN	O	O
of	NN	O	O
underlying	NN	O	O
immunologic	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

A	NN	O	O
51	NN	O	O
/	NN	O	O
2	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
black	NN	O	O
girl	NN	O	O
with	NN	O	O
recurrent	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
and	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
sixth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C6	NN	O	O
)	NN	O	O
is	NN	O	O
reported	NN	O	O
.	NN	O	O

To	NN	O	O
explore	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
recurrent	NN	O	O
neisserial	NN	O	B-Disease
infections	NN	O	I-Disease
in	NN	O	O
C6	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
detailed	NN	O	O
analysis	NN	O	O
of	NN	O	O
her	NN	O	O
immune	NN	O	O
competence	NN	O	O
was	NN	O	O
conducted	NN	O	O
.	NN	O	O

Her	NN	O	O
serum	NN	O	O
had	NN	O	O
normal	NN	O	O
chemotactic	NN	O	O
,	NN	O	O
opsonic	NN	O	O
,	NN	O	O
alternative	NN	O	O
complement	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
specific	NN	O	O
antibody	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
lacked	NN	O	O
complement	NN	O	O
-	NN	O	O
mediated	NN	O	O
bacteriolytic	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
her	NN	O	O
C6	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
serum	NN	O	O
was	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
normal	NN	O	O
serum	NN	O	O
in	NN	O	O
a	NN	O	O
complement	NN	O	O
-	NN	O	O
dependent	NN	O	O
assay	NN	O	O
of	NN	O	O
phagocyte	NN	O	O
bactericidal	NN	O	O
activity	NN	O	O
.	NN	O	O

Absent	NN	O	O
bacteriolysis	NN	O	O
remains	NN	O	O
the	NN	O	O
only	NN	O	O
consistent	NN	O	O
defect	NN	O	O
associated	NN	O	O
with	NN	O	O
recurrent	NN	O	O
neisserial	NN	O	B-Disease
infections	NN	O	I-Disease
and	NN	O	O
absence	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
late	NN	O	O
-	NN	O	O
acting	NN	O	O
complement	NN	O	O
components	NN	O	O
.	NN	O	O
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fifth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
human	NN	O	O
subjects	NN	O	O
.	NN	O	O

Clinical	NN	O	O
,	NN	O	O
genetic	NN	O	O
and	NN	O	O
immunologic	NN	O	O
studies	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
kindred	NN	O	O
.	NN	O	O

The	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
kindred	NN	O	O
with	NN	O	O
a	NN	O	O
heritable	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fifth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C5	NN	O	O
)	NN	O	O
has	NN	O	O
permitted	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
new	NN	O	O
clinical	NN	O	O
,	NN	O	O
genetic	NN	O	O
and	NN	O	O
immunologic	NN	O	O
data	NN	O	O
concerning	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
C5	NN	O	O
in	NN	O	O
human	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
,	NN	O	O
who	NN	O	O
has	NN	O	O
had	NN	O	O
nine	NN	O	O
episodes	NN	O	O
of	NN	O	O
disseminated	NN	O	O
gonococcal	NN	O	B-Disease
infection	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
a	NN	O	O
hemolytic	NN	O	O
C5	NN	O	O
level	NN	O	O
of	NN	O	O
approximately	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
per	NN	O	O
cent	NN	O	O
of	NN	O	O
normal	NN	O	O
.	NN	O	O

No	NN	O	O
C5	NN	O	O
protein	NN	O	O
was	NN	O	O
detectable	NN	O	O
,	NN	O	O
but	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
functional	NN	O	O
C5	NN	O	O
activity	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
using	NN	O	O
a	NN	O	O
sensitive	NN	O	O
bactericidal	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
probands	NN	O	O
twin	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
another	NN	O	O
sister	NN	O	O
also	NN	O	O
had	NN	O	O
extremely	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
hemolytic	NN	O	O
C5	NN	O	O
(	NN	O	O
approximately	NN	O	O
0	NN	O	O
.	NN	O	O
5	NN	O	O
per	NN	O	O
cent	NN	O	O
normal	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
both	NN	O	O
these	NN	O	O
subjects	NN	O	O
have	NN	O	O
been	NN	O	O
healthy	NN	O	O
.	NN	O	O

Hemolytic	NN	O	O
complement	NN	O	O
and	NN	O	O
bacteriolytic	NN	O	O
activity	NN	O	O
could	NN	O	O
be	NN	O	O
restored	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
purified	NN	O	O
C5	NN	O	O
.	NN	O	O

No	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
for	NN	O	O
polymorphonuclear	NN	O	O
leukocytes	NN	O	O
could	NN	O	O
be	NN	O	O
generated	NN	O	O
in	NN	O	O
the	NN	O	O
C5	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
serums	NN	O	O
upon	NN	O	O
activation	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
classic	NN	O	O
or	NN	O	O
alternative	NN	O	O
pathways	NN	O	O
,	NN	O	O
again	NN	O	O
demonstrating	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
C5	NN	O	O
in	NN	O	O
human	NN	O	O
subjects	NN	O	O
for	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
factors	NN	O	O
.	NN	O	O

The	NN	O	O
chemotactic	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
polymorphonuclear	NN	O	O
leukocytes	NN	O	O
and	NN	O	O
monocytes	NN	O	O
to	NN	O	O
preformed	NN	O	O
chemotactic	NN	O	O
factors	NN	O	O
was	NN	O	O
not	NN	O	O
depressed	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
two	NN	O	O
of	NN	O	O
32	NN	O	O
other	NN	O	O
family	NN	O	O
members	NN	O	O
from	NN	O	O
three	NN	O	O
generations	NN	O	O
had	NN	O	O
depressed	NN	O	O
whole	NN	O	O
hemolytic	NN	O	O
complement	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
19	NN	O	O
of	NN	O	O
30	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
hemolytic	NN	O	O
C5	NN	O	O
ranged	NN	O	O
from	NN	O	O
13	NN	O	O
to	NN	O	O
64	NN	O	O
per	NN	O	O
cent	NN	O	O
of	NN	O	O
normal	NN	O	O
.	NN	O	O

No	NN	O	O
linkage	NN	O	O
for	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
A	NN	O	O
or	NN	O	O
B	NN	O	O
loci	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
histocompatibility	NN	O	O
complex	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
an	NN	O	O
autosomal	NN	O	O
codominant	NN	O	O
mode	NN	O	O
of	NN	O	O
inheritance	NN	O	O
of	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
C5	NN	O	I-Disease
is	NN	O	O
compatible	NN	O	O
with	NN	O	O
good	NN	O	O
health	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
repeated	NN	O	O
disseminated	NN	O	O
gonococcal	NN	O	B-Disease
infection	NN	O	I-Disease

Incidence	NN	O	O
and	NN	O	O
characteristics	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variants	NN	O	O
in	NN	O	O
Japan	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
3000	NN	O	O
men	NN	O	O
living	NN	O	O
in	NN	O	O
Yamaguchi	NN	O	O
were	NN	O	O
screened	NN	O	O
for	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
using	NN	O	O
Beutlers	NN	O	O
spot	NN	O	O
test	NN	O	O
and	NN	O	O
three	NN	O	O
types	NN	O	O
of	NN	O	O
starch	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
.	NN	O	O

These	NN	O	O
electrophoresis	NN	O	O
used	NN	O	O
a	NN	O	O
phosphate	NN	O	O
buffer	NN	O	O
system	NN	O	O
at	NN	O	O
pH	NN	O	O
7	NN	O	O
.	NN	O	O

0	NN	O	O
,	NN	O	O
a	NN	O	O
TRIS	NN	O	O
-	NN	O	O
EDTA	NN	O	O
-	NN	O	O
borate	NN	O	O
buffer	NN	O	O
system	NN	O	O
at	NN	O	O
pH	NN	O	O
8	NN	O	O
.	NN	O	O

6	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
TRIS	NN	O	O
-	NN	O	O
hydrochloride	NN	O	O
buffer	NN	O	O
system	NN	O	O
at	NN	O	O
pH	NN	O	O
8	NN	O	O
.	NN	O	O
.	NN	O	O

Fifteen	NN	O	O
G6PD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
variants	NN	O	O
were	NN	O	O
found	NN	O	O
at	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
and	NN	O	O
classified	NN	O	O
into	NN	O	O
four	NN	O	O
groups	NN	O	O
.	NN	O	O

As	NN	O	O
new	NN	O	O
variants	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Konan	NN	O	O
,	NN	O	O
Kamiube	NN	O	O
,	NN	O	O
and	NN	O	O
Kiwa	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

These	NN	O	O
three	NN	O	O
variants	NN	O	O
had	NN	O	O
a	NN	O	O
mild	NN	O	O
to	NN	O	O
moderate	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
were	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
any	NN	O	O
clinical	NN	O	O
signs	NN	O	O
.	NN	O	O

G6PD	NN	O	O
Konan	NN	O	O
had	NN	O	O
fast	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
levels	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Kiwa	NN	O	O
had	NN	O	O
slightly	NN	O	O
elevated	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Kamiube	NN	O	O
had	NN	O	O
normal	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
.	NN	O	O

These	NN	O	O
three	NN	O	O
variants	NN	O	O
had	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
Km	NN	O	O
G6P	NN	O	O
,	NN	O	O
Km	NN	O	O
NADP	NN	O	O
,	NN	O	O
and	NN	O	O
Ki	NN	O	O
NADPH	NN	O	O
,	NN	O	O
normal	NN	O	O
utilizations	NN	O	O
of	NN	O	O
both	NN	O	O
2	NN	O	O
-	NN	O	O
deoxy	NN	O	O
-	NN	O	O
G6P	NN	O	O
and	NN	O	O
deamino	NN	O	O
-	NN	O	O
NAPD	NN	O	O
,	NN	O	O
normal	NN	O	O
heat	NN	O	O
stability	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
normal	NN	O	O
pH	NN	O	O
curve	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
variant	NN	O	O
was	NN	O	O
G6PD	NN	O	O
Ube	NN	O	O
,	NN	O	O
which	NN	O	O
we	NN	O	O
had	NN	O	O
previously	NN	O	O
found	NN	O	O
in	NN	O	O
Yamaguchi	NN	O	O
(	NN	O	O
Nakashima	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1977	NN	O	O
)	NN	O	O
.	NN	O	O

One	NN	O	O
boy	NN	O	O
with	NN	O	O
G6PD	NN	O	O
Ube	NN	O	O
was	NN	O	O
Korean	NN	O	O

Utilization	NN	O	O
of	NN	O	O
purines	NN	O	O
by	NN	O	O
an	NN	O	O
HPRT	NN	O	O
variant	NN	O	O
in	NN	O	O
an	NN	O	O
intelligent	NN	O	O
,	NN	O	O
nonmutilative	NN	O	O
patient	NN	O	O
with	NN	O	O
features	NN	O	O
of	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
patient	NN	O	O
,	NN	O	O
H	NN	O	O
.	NN	O	O

Chr	NN	O	O
.	NN	O	O

B	NN	O	O
.	NN	O	O
,	NN	O	O
was	NN	O	O
among	NN	O	O
the	NN	O	O
first	NN	O	O
reported	NN	O	O
with	NN	O	O
hyperuricemia	NN	O	B-Disease
and	NN	O	O
central	NN	O	B-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
symptoms	NN	O	I-Disease
.	NN	O	O

He	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
variant	NN	O	O
of	NN	O	O
hypoxanthine	NN	O	O
guanine	NN	O	O
phosphoribosyl	NN	O	O
transferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
;	NN	O	O
E	NN	O	O
.	NN	O	O
C	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O
4	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O
8	NN	O	O
)	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
enzyme	NN	O	O
present	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
patient	NN	O	O
had	NN	O	O
chroeoathetosis	NN	O	B-Disease
,	NN	O	O
spasticity	NN	O	B-Disease
,	NN	O	O
dysarthric	NN	O	B-Disease
speech	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hyperuricemia	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
his	NN	O	O
intelligence	NN	O	O
was	NN	O	O
normal	NN	O	O
and	NN	O	O
he	NN	O	O
had	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
self	NN	O	O
-	NN	O	O
mutilation	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
activity	NN	O	O
of	NN	O	O
HPRT	NN	O	O
in	NN	O	O
the	NN	O	O
lysates	NN	O	O
of	NN	O	O
erythrocytes	NN	O	O
and	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
when	NN	O	O
analyzed	NN	O	O
in	NN	O	O
the	NN	O	O
usual	NN	O	O
manner	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
newly	NN	O	O
developed	NN	O	O
method	NN	O	O
for	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
purine	NN	O	O
metabolism	NN	O	O
in	NN	O	O
intact	NN	O	O
cultured	NN	O	O
cells	NN	O	O
,	NN	O	O
this	NN	O	O
patient	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
metabolize	NN	O	O
some	NN	O	O
9	NN	O	O
%	NN	O	O
of	NN	O	O
8	NN	O	O
-	NN	O	O
14C	NN	O	O
-	NN	O	O
hypoxanthine	NN	O	O
,	NN	O	O
and	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
isotope	NN	O	O
utilized	NN	O	O
was	NN	O	O
converted	NN	O	O
to	NN	O	O
adenine	NN	O	O
and	NN	O	O
guanine	NN	O	O
nucleotides	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
cells	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
were	NN	O	O
virtually	NN	O	O
completely	NN	O	O
unable	NN	O	O
to	NN	O	O
convert	NN	O	O
hypoxanthine	NN	O	O
to	NN	O	O
nucleotides	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
fibroblasts	NN	O	O
were	NN	O	O
even	NN	O	O
more	NN	O	O
efficient	NN	O	O
in	NN	O	O
the	NN	O	O
metabolism	NN	O	O
of	NN	O	O
8	NN	O	O
-	NN	O	O
14C	NN	O	O
-	NN	O	O
guanine	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
utilized	NN	O	O
to	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
27	NN	O	O
%	NN	O	O
,	NN	O	O
over	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
which	NN	O	O
was	NN	O	O
converted	NN	O	O
to	NN	O	O
guanine	NN	O	O
and	NN	O	O
adenine	NN	O	O
nucleotides	NN	O	O
.	NN	O	O

The	NN	O	O
growth	NN	O	O
of	NN	O	O
the	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
of	NN	O	O
this	NN	O	O
patient	NN	O	O
was	NN	O	O
intermediate	NN	O	O
in	NN	O	O
media	NN	O	O
containing	NN	O	O
hypoxanthine	NN	O	O
aminopterin	NN	O	O
thymidine	NN	O	O
(	NN	O	O
HAT	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
cells	NN	O	O
was	NN	O	O
inhibited	NN	O	O
and	NN	O	O
normal	NN	O	O
cells	NN	O	O
grew	NN	O	O
normally	NN	O	O
.	NN	O	O

Similarly	NN	O	O
in	NN	O	O
8	NN	O	O
-	NN	O	O
azaguanine	NN	O	O
,	NN	O	O
6	NN	O	O
-	NN	O	O
thioguanine	NN	O	O
,	NN	O	O
and	NN	O	O
8	NN	O	O
-	NN	O	O
azahypoxanthine	NN	O	O
,	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
cells	NN	O	O
was	NN	O	O
intermediate	NN	O	O
between	NN	O	O
normal	NN	O	O
and	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
cells	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
disorders	NN	O	O
in	NN	O	O
purine	NN	O	O
metabolism	NN	O	O
involving	NN	O	O
the	NN	O	O
HPRT	NN	O	O
gene	NN	O	O
.	NN	O	O

They	NN	O	O
document	NN	O	O
that	NN	O	O
this	NN	O	O
famous	NN	O	O
patient	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
the	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease

Localisation	NN	O	O
of	NN	O	O
the	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
on	NN	O	O
the	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
by	NN	O	O
linkage	NN	O	O
to	NN	O	O
cloned	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
.	NN	O	O

A	NN	O	O
linkage	NN	O	O
study	NN	O	O
in	NN	O	O
30	NN	O	O
Becker	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
BMD	NN	O	B-Disease
)	NN	O	O
kindreds	NN	O	O
using	NN	O	O
three	NN	O	O
cloned	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
which	NN	O	O
demonstrate	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
RFLPs	NN	O	O
)	NN	O	O
,	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
BMD	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
p21	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
genes	NN	O	O
for	NN	O	O
Becker	NN	O	B-Disease
and	NN	O	I-Disease
Duchenne	NN	O	I-Disease
dystrophies	NN	O	I-Disease
must	NN	O	O
therefore	NN	O	O
be	NN	O	O
closely	NN	O	O
linked	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
allelic	NN	O	O
,	NN	O	O
and	NN	O	O
any	NN	O	O
future	NN	O	O
DNA	NN	O	O
probes	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
practical	NN	O	O
use	NN	O	O
in	NN	O	O
one	NN	O	O
disorder	NN	O	O
should	NN	O	O
be	NN	O	O
equally	NN	O	O
applicable	NN	O	O
to	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

The	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
also	NN	O	O
provides	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
recombination	NN	O	O
along	NN	O	O
the	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
and	NN	O	O
across	NN	O	O
the	NN	O	O
centromeric	NN	O	O
region	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
evidence	NN	O	O
for	NN	O	O
new	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
hprt	NN	O	O
locus	NN	O	O
in	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Hypoxanthine	NN	O	O
-	NN	O	O
guanine	NN	O	O
phosphoribosyltransferase	NN	O	O
(	NN	O	O
HPRT	NN	O	O
;	NN	O	O
EC2	NN	O	O
.	NN	O	O
4	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O
8	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
metabolic	NN	O	O
salvage	NN	O	O
of	NN	O	O
purines	NN	O	O
,	NN	O	O
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
an	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
gene	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Partial	NN	O	O
HPRT	NN	O	B-Disease
deficiencies	NN	O	I-Disease
are	NN	O	O
associated	NN	O	O
with	NN	O	O
gouty	NN	O	B-Disease
arthritis	NN	O	I-Disease
,	NN	O	O
while	NN	O	O
absence	NN	O	O
of	NN	O	O
activity	NN	O	O
results	NN	O	O
in	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
L	NN	O	B-Disease
-	NN	O	I-Disease
N	NN	O	I-Disease
)	NN	O	O
.	NN	O	O

L	NN	O	B-Disease
-	NN	O	I-Disease
N	NN	O	I-Disease
patients	NN	O	O
fail	NN	O	O
to	NN	O	O
reproduce	NN	O	O
and	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
state	NN	O	O
appears	NN	O	O
to	NN	O	O
confer	NN	O	O
no	NN	O	O
selective	NN	O	O
advantage	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Haldanes	NN	O	O
principle	NN	O	O
predicts	NN	O	O
that	NN	O	O
new	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
hprt	NN	O	O
locus	NN	O	O
must	NN	O	O
occur	NN	O	O
frequently	NN	O	O
in	NN	O	O
order	NN	O	O
for	NN	O	O
L	NN	O	B-Disease
-	NN	O	I-Disease
N	NN	O	I-Disease
syndrome	NN	O	I-Disease
to	NN	O	O
be	NN	O	O
maintained	NN	O	O
in	NN	O	O
the	NN	O	O
population	NN	O	O
.	NN	O	O

This	NN	O	O
constant	NN	O	O
introduction	NN	O	O
of	NN	O	O
new	NN	O	O
mutations	NN	O	O
would	NN	O	O
be	NN	O	O
expected	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
collection	NN	O	O
of	NN	O	O
genetic	NN	O	B-Disease
lesions	NN	O	I-Disease
,	NN	O	O
some	NN	O	O
of	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
novel	NN	O	O
.	NN	O	O

As	NN	O	O
we	NN	O	O
report	NN	O	O
here	NN	O	O
,	NN	O	O
the	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
hprt	NN	O	O
gene	NN	O	O
of	NN	O	O
seven	NN	O	O
L	NN	O	B-Disease
-	NN	O	I-Disease
N	NN	O	I-Disease
patients	NN	O	O
,	NN	O	O
selected	NN	O	O
from	NN	O	O
an	NN	O	O
initial	NN	O	O
survey	NN	O	O
of	NN	O	O
28	NN	O	O
patients	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
and	NN	O	O
all	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
distinctly	NN	O	O
different	NN	O	O
,	NN	O	O
as	NN	O	O
predicted	NN	O	O
.	NN	O	O

The	NN	O	O
origin	NN	O	O
of	NN	O	O
one	NN	O	O
unusual	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
from	NN	O	O
four	NN	O	O
generations	NN	O	O
of	NN	O	O
family	NN	O	O
members	NN	O	O
.	NN	O	O

Further	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
new	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
hprt	NN	O	O
locus	NN	O	O
should	NN	O	O
aid	NN	O	O
in	NN	O	O
resolving	NN	O	O
the	NN	O	O
issue	NN	O	O
of	NN	O	O
an	NN	O	O
apparent	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
hprt	NN	O	O
mutations	NN	O	O
in	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O

Allelic	NN	O	O
exclusion	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
in	NN	O	O
platelets	NN	O	O
and	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
a	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
carrier	NN	O	O
.	NN	O	O

An	NN	O	O
obligate	NN	O	O
carrier	NN	O	O
of	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
who	NN	O	O
was	NN	O	O
also	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
A	NN	O	O
and	NN	O	O
B	NN	O	O
types	NN	O	O
of	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
was	NN	O	O
found	NN	O	O
.	NN	O	O

With	NN	O	O
her	NN	O	O
it	NN	O	O
became	NN	O	O
possible	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
allelic	NN	O	O
exclusion	NN	O	O
occurs	NN	O	O
in	NN	O	O
particular	NN	O	O
cell	NN	O	O
-	NN	O	O
types	NN	O	O
of	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
carrier	NN	O	O
.	NN	O	O

If	NN	O	O
so	NN	O	O
,	NN	O	O
the	NN	O	O
remaining	NN	O	O
cells	NN	O	O
of	NN	O	O
a	NN	O	O
particular	NN	O	O
cell	NN	O	O
-	NN	O	O
type	NN	O	O
would	NN	O	O
express	NN	O	O
only	NN	O	O
the	NN	O	O
normal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
and	NN	O	O
only	NN	O	O
one	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
isoenzyme	NN	O	O
would	NN	O	O
be	NN	O	O
demonstrable	NN	O	O
.	NN	O	O

This	NN	O	O
carrier	NN	O	O
had	NN	O	O
only	NN	O	O
the	NN	O	O
B	NN	O	O
isoenzyme	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
in	NN	O	O
platelets	NN	O	O
and	NN	O	O
thymus	NN	O	O
-	NN	O	O
derived	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
although	NN	O	O
both	NN	O	O
isoenzymes	NN	O	O
A	NN	O	O
and	NN	O	O
B	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
erythrocytes	NN	O	O
and	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
selection	NN	O	O
against	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
occurs	NN	O	O
in	NN	O	O
platelets	NN	O	O
and	NN	O	O
thymus	NN	O	O
-	NN	O	O
derived	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
defects	NN	O	O
associated	NN	O	O
with	NN	O	O
WAS	NN	O	B-Disease
expressed	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
-	NN	O	O
types	NN	O	O
may	NN	O	O
be	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
genesis	NN	O	O
of	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Complement	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
nephritis	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
report	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
is	NN	O	O
described	NN	O	O
in	NN	O	O
which	NN	O	O
three	NN	O	O
children	NN	O	O
had	NN	O	O
homozygous	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
C3	NN	O	I-Disease
and	NN	O	O
in	NN	O	O
which	NN	O	O
both	NN	O	O
parents	NN	O	O
and	NN	O	O
two	NN	O	O
other	NN	O	O
children	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
C3	NN	O	O
null	NN	O	O
gene	NN	O	O
.	NN	O	O

One	NN	O	O
child	NN	O	O
with	NN	O	O
heterozygous	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
membranoproliferative	NN	O	O
glomerulonephritis	NN	O	B-Disease
;	NN	O	O
proteinuria	NN	O	B-Disease
and	NN	O	O
/	NN	O	O
or	NN	O	O
microscopical	NN	O	O
haematuria	NN	O	B-Disease
was	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
homozygous	NN	O	O
C3	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
children	NN	O	O
.	NN	O	O

All	NN	O	O
children	NN	O	O
with	NN	O	O
homozygous	NN	O	O
or	NN	O	O
heterozygous	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
were	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
varying	NN	O	O
degree	NN	O	O
,	NN	O	O
susceptible	NN	O	O
to	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
only	NN	O	O
child	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
with	NN	O	O
normal	NN	O	O
complement	NN	O	O
had	NN	O	O
no	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
infection	NN	O	O
and	NN	O	O
no	NN	O	O
renal	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
family	NN	O	O
study	NN	O	O
provides	NN	O	O
further	NN	O	O
support	NN	O	O
for	NN	O	O
the	NN	O	O
proposal	NN	O	O
that	NN	O	O
C3	NN	O	B-Disease
deficiency	NN	O	I-Disease
predisposes	NN	O	O
to	NN	O	O
nephritis	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Heterogeneity	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Algeria	NN	O	O
.	NN	O	O

Study	NN	O	O
in	NN	O	O
Northern	NN	O	O
Algeria	NN	O	O
with	NN	O	O
description	NN	O	O
of	NN	O	O
five	NN	O	O
new	NN	O	O
variants	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
found	NN	O	O
in	NN	O	O
3	NN	O	O
.	NN	O	O

2	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
male	NN	O	O
population	NN	O	O
living	NN	O	O
in	NN	O	O
the	NN	O	O
urban	NN	O	O
area	NN	O	O
of	NN	O	O
Algiers	NN	O	O
.	NN	O	O

The	NN	O	O
deficient	NN	O	O
subjects	NN	O	O
originated	NN	O	O
from	NN	O	O
multiple	NN	O	O
geographic	NN	O	O
regions	NN	O	O
of	NN	O	O
Northern	NN	O	O
Algeria	NN	O	O
,	NN	O	O
with	NN	O	O
prevalence	NN	O	O
of	NN	O	O
individuals	NN	O	O
of	NN	O	O
Berber	NN	O	O
-	NN	O	O
Kabyle	NN	O	O
origin	NN	O	O
.	NN	O	O

Red	NN	O	O
blood	NN	O	O
cell	NN	O	O
G6PD	NN	O	O
was	NN	O	O
partially	NN	O	O
purified	NN	O	O
and	NN	O	O
characterized	NN	O	O
in	NN	O	O
deficient	NN	O	O
males	NN	O	O
from	NN	O	O
17	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
six	NN	O	O
different	NN	O	O
variants	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

Among	NN	O	O
them	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
,	NN	O	O
the	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Kabyle	NN	O	O
variant	NN	O	O
,	NN	O	O
had	NN	O	O
been	NN	O	O
previously	NN	O	O
described	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
nine	NN	O	O
families	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
five	NN	O	O
variants	NN	O	O
were	NN	O	O
new	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Laghouat	NN	O	O
(	NN	O	O
four	NN	O	O
cases	NN	O	O
)	NN	O	O
,	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Blida	NN	O	O
(	NN	O	O
one	NN	O	O
case	NN	O	O
)	NN	O	O
,	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Thenia	NN	O	O
(	NN	O	O
one	NN	O	O
case	NN	O	O
)	NN	O	O
,	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Titteri	NN	O	O
(	NN	O	O
one	NN	O	O
case	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
Gd	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
Alger	NN	O	O
(	NN	O	O
two	NN	O	O
brothers	NN	O	O
)	NN	O	O
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
the	NN	O	O
common	NN	O	O
Mediterranean	NN	O	O
variant	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
.	NN	O	O

G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
heterogeneous	NN	O	O
in	NN	O	O
northern	NN	O	O
Algeria	NN	O	O
where	NN	O	O
autochtonous	NN	O	O
variants	NN	O	O
seem	NN	O	O
to	NN	O	O
prevail	NN	O	O
.	NN	O	O

The	NN	O	O
Kabyle	NN	O	O
variant	NN	O	O
may	NN	O	O
be	NN	O	O
common	NN	O	O
in	NN	O	O
this	NN	O	O
country	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
inhibits	NN	O	O
in	NN	O	O
vitro	NN	O	O
growth	NN	O	O
of	NN	O	O
Plasmodium	NN	O	O
falciparum	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
;	NN	O	I-Disease
EC	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
1	NN	O	I-Disease
.	NN	O	I-Disease
49	NN	O	I-Disease
)	NN	O	I-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
red	NN	O	O
blood	NN	O	O
cells	NN	O	O
from	NN	O	O
male	NN	O	O
hemizygotes	NN	O	O
and	NN	O	O
female	NN	O	O
heterozygotes	NN	O	O
from	NN	O	O
the	NN	O	O
island	NN	O	O
of	NN	O	O
Sardinia	NN	O	O
were	NN	O	O
studied	NN	O	O
for	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
support	NN	O	O
growth	NN	O	O
in	NN	O	O
vitro	NN	O	O
of	NN	O	O
the	NN	O	O
malaria	NN	O	B-Disease
-	NN	O	O
causing	NN	O	O
organism	NN	O	O
Plasmodium	NN	O	O
falciparum	NN	O	O
.	NN	O	O

Parasite	NN	O	O
growth	NN	O	O
was	NN	O	O
approximately	NN	O	O
one	NN	O	O
-	NN	O	O
third	NN	O	O
of	NN	O	O
normal	NN	O	O
in	NN	O	O
both	NN	O	O
hemi	NN	O	O
-	NN	O	O
and	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
Sardinians	NN	O	O
with	NN	O	O
the	NN	O	O
beta	NN	O	O
0	NN	O	O
-	NN	O	O
thalassemia	NN	O	O
trait	NN	O	O
,	NN	O	O
parasite	NN	O	O
growth	NN	O	O
was	NN	O	O
normal	NN	O	O
except	NN	O	O
when	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
occurred	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
thalassemia	NN	O	B-Disease
trait	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
may	NN	O	O
confer	NN	O	O
a	NN	O	O
selective	NN	O	O
advantage	NN	O	O
in	NN	O	O
a	NN	O	O
malarious	NN	O	B-Disease
area	NN	O	O
;	NN	O	O
the	NN	O	O
female	NN	O	O
heterozygote	NN	O	O
may	NN	O	O
be	NN	O	O
at	NN	O	O
a	NN	O	O
particular	NN	O	O
advantage	NN	O	O
because	NN	O	O
resistance	NN	O	O
to	NN	O	O
malaria	NN	O	B-Disease
equals	NN	O	O
that	NN	O	O
of	NN	O	O
male	NN	O	O
hemizygotes	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
fatal	NN	O	B-Disease
hemolysis	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
less	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
more	NN	O	O
female	NN	O	O
heterozygotes	NN	O	O
must	NN	O	O
be	NN	O	O
studied	NN	O	O
to	NN	O	O
confirm	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
.	NN	O	O

No	NN	O	O
protective	NN	O	O
effect	NN	O	O
of	NN	O	O
beta	NN	O	O
0	NN	O	O
-	NN	O	O
thalassemia	NN	O	O
trait	NN	O	O
could	NN	O	O
be	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Bone	NN	O	O
marrow	NN	O	O
transplant	NN	O	O
in	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
.	NN	O	O

An	NN	O	O
allogeneic	NN	O	O
bone	NN	O	O
marrow	NN	O	O
transplant	NN	O	O
(	NN	O	O
BMT	NN	O	O
)	NN	O	O
from	NN	O	O
a	NN	O	O
normal	NN	O	O
HLA	NN	O	O
identical	NN	O	O
sibling	NN	O	O
donor	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
a	NN	O	O
13	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
boy	NN	O	O
with	NN	O	O
rapidly	NN	O	O
progressive	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Engraftment	NN	O	O
and	NN	O	O
complete	NN	O	O
hematologic	NN	O	O
recovery	NN	O	O
occurred	NN	O	O
within	NN	O	O
4	NN	O	O
weeks	NN	O	O
,	NN	O	O
but	NN	O	O
neurologic	NN	O	B-Disease
deterioration	NN	O	I-Disease
continued	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
died	NN	O	O
of	NN	O	O
an	NN	O	O
adenovirus	NN	O	B-Disease
infection	NN	O	I-Disease
141	NN	O	O
days	NN	O	O
after	NN	O	O
BMT	NN	O	O
.	NN	O	O

ALD	NN	O	B-Disease
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
abnormally	NN	O	O
high	NN	O	O
plasma	NN	O	O
levels	NN	O	O
of	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
(	NN	O	O
VLCFA	NN	O	O
)	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
impaired	NN	O	O
capacity	NN	O	O
to	NN	O	O
degrade	NN	O	O
them	NN	O	O
.	NN	O	O

Ten	NN	O	O
days	NN	O	O
after	NN	O	O
BMT	NN	O	O
,	NN	O	O
the	NN	O	O
white	NN	O	O
blood	NN	O	O
cell	NN	O	O
VLCFA	NN	O	O
levels	NN	O	O
and	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
became	NN	O	O
normal	NN	O	O
;	NN	O	O
after	NN	O	O
3	NN	O	O
months	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
progressive	NN	O	O
reduction	NN	O	O
of	NN	O	O
plasma	NN	O	O
VLCFA	NN	O	O
to	NN	O	O
levels	NN	O	O
only	NN	O	O
slightly	NN	O	O
above	NN	O	O
normal	NN	O	O
.	NN	O	O
.	NN	O	O

[	NN	O	O
Gd	NN	O	O
-	NN	O	O
allele	NN	O	O
distribution	NN	O	O
patterns	NN	O	O
in	NN	O	O
Azerbaijan	NN	O	O
.	NN	O	O
III	NN	O	O
.	NN	O	O
The	NN	O	O
identification	NN	O	O
of	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
]	NN	O	O

In	NN	O	O
28	NN	O	O
families	NN	O	O
with	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
living	NN	O	O
in	NN	O	O
3	NN	O	O
settlements	NN	O	O
of	NN	O	O
Shekii	NN	O	O
district	NN	O	O
of	NN	O	O
Azerbaijan	NN	O	O
11	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
of	NN	O	O
II	NN	O	O
and	NN	O	O
III	NN	O	O
classes	NN	O	O
differing	NN	O	O
by	NN	O	O
kinetic	NN	O	O
properties	NN	O	O
were	NN	O	O
identified	NN	O	O
according	NN	O	O
WHO	NN	O	O
program	NN	O	O
.	NN	O	O

9	NN	O	O
of	NN	O	O
them	NN	O	O
are	NN	O	O
characterized	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
G6PD	NN	O	O
spectra	NN	O	O
in	NN	O	O
two	NN	O	O
subpopulations	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
mixed	NN	O	O
group	NN	O	O
permits	NN	O	O
to	NN	O	O
make	NN	O	O
a	NN	O	O
conclusion	NN	O	O
about	NN	O	O
existence	NN	O	O
of	NN	O	O
common	NN	O	O
and	NN	O	O
rare	NN	O	O
G6PD	NN	O	O
alleles	NN	O	O
in	NN	O	O
examined	NN	O	O
population	NN	O	O
.	NN	O	O

They	NN	O	O
distribute	NN	O	O
by	NN	O	O
gene	NN	O	O
drift	NN	O	O
supported	NN	O	O
by	NN	O	O
natural	NN	O	O
selection	NN	O	O
.	NN	O	O

Among	NN	O	O
7	NN	O	O
samples	NN	O	O
of	NN	O	O
G6PD	NN	O	O
with	NN	O	O
normal	NN	O	O
and	NN	O	O
increased	NN	O	O
activity	NN	O	O
two	NN	O	O
new	NN	O	O
variants	NN	O	O
of	NN	O	O
IV	NN	O	O
class	NN	O	O
-	NN	O	O
-	NN	O	O
Nukha	NN	O	O
and	NN	O	O
Bash	NN	O	O
-	NN	O	O
Kungut	NN	O	O
-	NN	O	O
-	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
:	NN	O	O
survey	NN	O	O
of	NN	O	O
303	NN	O	O
cases	NN	O	O
:	NN	O	O
biochemistry	NN	O	O
,	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
and	NN	O	O
therapy	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
genetically	NN	O	O
determined	NN	O	O
disorder	NN	O	O
associated	NN	O	O
with	NN	O	O
progressive	NN	O	B-Disease
central	NN	O	I-Disease
demyelination	NN	O	I-Disease
and	NN	O	O
adrenal	NN	O	B-Disease
cortical	NN	O	I-Disease
insufficiency	NN	O	I-Disease
.	NN	O	O

All	NN	O	O
affected	NN	O	O
persons	NN	O	O
show	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
saturated	NN	O	O
unbranched	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
particularly	NN	O	O
hexacosanoate	NN	O	O
(	NN	O	O
C26	NN	O	O
0	NN	O	O
)	NN	O	O
,	NN	O	O
because	NN	O	O
of	NN	O	O
impaired	NN	O	O
capacity	NN	O	O
to	NN	O	O
degrade	NN	O	O
these	NN	O	O
acids	NN	O	O
.	NN	O	O

This	NN	O	O
degradation	NN	O	O
normally	NN	O	O
takes	NN	O	O
place	NN	O	O
in	NN	O	O
a	NN	O	O
subcellular	NN	O	O
organelle	NN	O	O
called	NN	O	O
the	NN	O	O
peroxisome	NN	O	O
,	NN	O	O
and	NN	O	O
ALD	NN	O	B-Disease
,	NN	O	O
together	NN	O	O
with	NN	O	O
Zellwegers	NN	O	B-Disease
cerebrohepatorenal	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
now	NN	O	O
considered	NN	O	O
to	NN	O	O
belong	NN	O	O
to	NN	O	O
the	NN	O	O
newly	NN	O	O
formed	NN	O	O
category	NN	O	O
of	NN	O	O
peroxisomal	NN	O	B-Disease
disorders	NN	O	I-Disease
.	NN	O	O

Biochemical	NN	O	O
assays	NN	O	O
permit	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
identification	NN	O	O
of	NN	O	O
most	NN	O	O
heterozygotes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
303	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
in	NN	O	O
217	NN	O	O
kindreds	NN	O	O
.	NN	O	O

These	NN	O	O
patients	NN	O	O
show	NN	O	O
a	NN	O	O
wide	NN	O	O
phenotypic	NN	O	O
variation	NN	O	O
.	NN	O	O

Sixty	NN	O	O
percent	NN	O	O
of	NN	O	O
patients	NN	O	O
had	NN	O	O
childhood	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
17	NN	O	O
%	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
gene	NN	O	O
mapped	NN	O	O
to	NN	O	O
Xq28	NN	O	O
.	NN	O	O

Neonatal	NN	O	B-Disease
ALD	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
distinct	NN	O	O
entity	NN	O	O
with	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
inheritance	NN	O	O
and	NN	O	O
points	NN	O	O
of	NN	O	O
resemblance	NN	O	O
to	NN	O	O
Zellwegers	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
accounted	NN	O	O
for	NN	O	O
7	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cases	NN	O	O
.	NN	O	O

Although	NN	O	O
excess	NN	O	O
C26	NN	O	O
0	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
is	NN	O	O
partially	NN	O	O
of	NN	O	O
dietary	NN	O	O
origin	NN	O	O
,	NN	O	O
dietary	NN	O	O
C26	NN	O	O
0	NN	O	O
restriction	NN	O	O
did	NN	O	O
not	NN	O	O
produce	NN	O	O
clear	NN	O	O
benefit	NN	O	O
.	NN	O	O

Bone	NN	O	O
marrow	NN	O	O
transplant	NN	O	O
lowered	NN	O	O
the	NN	O	O
plasma	NN	O	O
C26	NN	O	O
0	NN	O	O
level	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
arrest	NN	O	O
neurological	NN	O	O
progression	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
polymorphism	NN	O	O
of	NN	O	O
G6PD	NN	O	O
in	NN	O	O
a	NN	O	O
Bulgarian	NN	O	O
population	NN	O	O
.	NN	O	O

Considerable	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
G6PD	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
Bulgarian	NN	O	O
population	NN	O	O
-	NN	O	O
14	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
isolated	NN	O	O
from	NN	O	O
117	NN	O	O
hemizygous	NN	O	O
carriers	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
type	NN	O	O
was	NN	O	O
a	NN	O	O
polymorphic	NN	O	O
variant	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Corinth	NN	O	O
occurred	NN	O	O
with	NN	O	O
high	NN	O	O
frequency	NN	O	O
.	NN	O	O

Two	NN	O	O
new	NN	O	O
variants	NN	O	O
were	NN	O	O
identified	NN	O	O
-	NN	O	O
G6PD	NN	O	O
Rudosem	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Nedelino	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
selected	NN	O	O
group	NN	O	O
of	NN	O	O
78	NN	O	O
subjects	NN	O	O
with	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
,	NN	O	O
four	NN	O	O
variants	NN	O	O
were	NN	O	O
established	NN	O	O
G6PD	NN	O	O
Mediterranian	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Corinth	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Seattle	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Ohut	NN	O	O
II	NN	O	O
.	NN	O	O
.	NN	O	O

Clinical	NN	O	O
use	NN	O	O
of	NN	O	O
DNA	NN	O	O
markers	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Seventy	NN	O	O
families	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
known	NN	O	O
to	NN	O	O
the	NN	O	O
Institute	NN	O	O
of	NN	O	O
Child	NN	O	O
Health	NN	O	O
fall	NN	O	O
into	NN	O	O
three	NN	O	O
categories	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
potential	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
DNA	NN	O	O
markers	NN	O	O
RC8	NN	O	O
and	NN	O	O
L1	NN	O	O
.	NN	O	O

28	NN	O	O
that	NN	O	O
bridge	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Families	NN	O	O
in	NN	O	O
which	NN	O	O
there	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
obligatory	NN	O	O
female	NN	O	O
heterozygote	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
13	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
prediction	NN	O	O
and	NN	O	O
exclusion	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
transmission	NN	O	O
may	NN	O	O
be	NN	O	O
possible	NN	O	O
,	NN	O	O
the	NN	O	O
accuracy	NN	O	O
being	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
closeness	NN	O	O
of	NN	O	O
the	NN	O	O
linkage	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
marker	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
to	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
;	NN	O	O
an	NN	O	O
illustrative	NN	O	O
case	NN	O	O
is	NN	O	O
reported	NN	O	O
.	NN	O	O

Families	NN	O	O
in	NN	O	O
which	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
single	NN	O	O
affected	NN	O	O
boy	NN	O	O
,	NN	O	O
who	NN	O	O
also	NN	O	O
has	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
healthy	NN	O	O
brothers	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
26	NN	O	O
)	NN	O	O
.	NN	O	O

Given	NN	O	O
an	NN	O	O
informative	NN	O	O
restriction	NN	O	O
fragment	NN	O	O
length	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
RFLP	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
probability	NN	O	O
that	NN	O	O
the	NN	O	O
boy	NN	O	O
represents	NN	O	O
a	NN	O	O
new	NN	O	O
mutation	NN	O	O
can	NN	O	O
be	NN	O	O
reassessed	NN	O	O
;	NN	O	O
it	NN	O	O
is	NN	O	O
also	NN	O	O
possible	NN	O	O
to	NN	O	O
exclude	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
a	NN	O	O
sister	NN	O	O
.	NN	O	O

Families	NN	O	O
with	NN	O	O
a	NN	O	O
single	NN	O	O
affected	NN	O	O
boy	NN	O	O
with	NN	O	O
no	NN	O	O
brother	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
30	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
exclusion	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
a	NN	O	O
sister	NN	O	O
may	NN	O	O
be	NN	O	O
possible	NN	O	O
.	NN	O	O

Only	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
was	NN	O	O
there	NN	O	O
no	NN	O	O
possibility	NN	O	O
of	NN	O	O
useful	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
required	NN	O	O
is	NN	O	O
described	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
need	NN	O	O
to	NN	O	O
check	NN	O	O
DMD	NN	O	B-Disease
families	NN	O	O
for	NN	O	O
informative	NN	O	O
RFLPs	NN	O	O
is	NN	O	O
stressed	NN	O	O
.	NN	O	O

Family	NN	O	O
studies	NN	O	O
in	NN	O	O
Bechterew	NN	O	B-Disease
'	NN	O	I-Disease
s	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
)	NN	O	O
III	NN	O	O
.	NN	O	O

Genetics	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
segregation	NN	O	O
analyses	NN	O	O
in	NN	O	O
75	NN	O	O
families	NN	O	O
where	NN	O	O
the	NN	O	O
proband	NN	O	O
had	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
,	NN	O	O
are	NN	O	O
presented	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
278	NN	O	O
adult	NN	O	O
,	NN	O	O
living	NN	O	O
first	NN	O	O
degree	NN	O	O
relatives	NN	O	O
,	NN	O	O
approximately	NN	O	O
85	NN	O	O
%	NN	O	O
cooperated	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
radiographical	NN	O	O
examinations	NN	O	O
were	NN	O	O
performed	NN	O	O
and	NN	O	O
HLA	NN	O	O
typing	NN	O	O
was	NN	O	O
conducted	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
were	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
our	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
syndrome	NN	O	O
where	NN	O	O
different	NN	O	O
genetic	NN	O	O
factors	NN	O	O
interact	NN	O	O
.	NN	O	O

Such	NN	O	O
known	NN	O	O
factors	NN	O	O
are	NN	O	O
HLA	NN	O	O
B27	NN	O	O
associated	NN	O	O
disease	NN	O	O
susceptibility	NN	O	O
,	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
psoriatic	NN	O	B-Disease
arthropathy	NN	O	I-Disease
and	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
entero	NN	O	B-Disease
-	NN	O	I-Disease
arthropathy	NN	O	I-Disease
.	NN	O	O

Radiographical	NN	O	O
sacro	NN	O	B-Disease
-	NN	O	I-Disease
iliitis	NN	O	I-Disease
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
HLA	NN	O	O
B27	NN	O	O
positive	NN	O	O
relatives	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
more	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
relatives	NN	O	O
to	NN	O	O
probands	NN	O	O
with	NN	O	O
psoriasis	NN	O	B-Disease
than	NN	O	O
in	NN	O	O
relatives	NN	O	O
to	NN	O	O
probands	NN	O	O
without	NN	O	O
psoriasis	NN	O	B-Disease
.	NN	O	O

Environmental	NN	O	O
factors	NN	O	O
(	NN	O	O
intestinal	NN	O	O
bacteria	NN	O	O
)	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
trigger	NN	O	O
the	NN	O	O
disease	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
some	NN	O	O
persons	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
postulated	NN	O	O
that	NN	O	O
all	NN	O	O
or	NN	O	O
most	NN	O	O
of	NN	O	O
them	NN	O	O
have	NN	O	O
the	NN	O	O
predisposition	NN	O	O
to	NN	O	O
develop	NN	O	O
disease	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
syndrome	NN	O	O
has	NN	O	O
a	NN	O	O
multifactorial	NN	O	O
etiology	NN	O	O
.	NN	O	O

The	NN	O	O
phenotypic	NN	O	O
expressions	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
genetic	NN	O	O
predispositions	NN	O	O
involved	NN	O	O
,	NN	O	O
include	NN	O	O
sacro	NN	O	B-Disease
-	NN	O	I-Disease
iliitis	NN	O	I-Disease
,	NN	O	O
psoriasis	NN	O	B-Disease
,	NN	O	O
acute	NN	O	B-Disease
anterior	NN	O	I-Disease
uveitis	NN	O	I-Disease
,	NN	O	O
peripheral	NN	O	O
arthropathy	NN	O	B-Disease
and	NN	O	O
inflammatory	NN	O	B-Disease
bowel	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
the	NN	O	O
descriptive	NN	O	O
name	NN	O	O
HEREDITARY	NN	O	B-Disease
MULTIFOCAL	NN	O	I-Disease
RELAPSING	NN	O	I-Disease
INFLAMMATION	NN	O	I-Disease
(	NN	O	O
HEMRI	NN	O	B-Disease
)	NN	O	O
for	NN	O	O
this	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Ankylosing	NN	O	B-Disease
spondylitis	NN	O	I-Disease
,	NN	O	O
psoriatic	NN	O	B-Disease
arthropathy	NN	O	I-Disease
and	NN	O	O
entero	NN	O	B-Disease
-	NN	O	I-Disease
arthropathy	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
regarded	NN	O	O
as	NN	O	O
clinical	NN	O	O
sub	NN	O	O
-	NN	O	O
types	NN	O	O
of	NN	O	O
the	NN	O	O
syndrome	NN	O	O
.	NN	O	O
.	NN	O	O

Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
chromosome	NN	O	O
15	NN	O	O
.	NN	O	O

A	NN	O	O
clinical	NN	O	O
discussion	NN	O	O
of	NN	O	O
20	NN	O	O
cases	NN	O	O
.	NN	O	O

A	NN	O	O
chromosome	NN	O	B-Disease
15	NN	O	I-Disease
anomaly	NN	O	I-Disease
was	NN	O	O
observed	NN	O	O
in	NN	O	O
12	NN	O	O
of	NN	O	O
20	NN	O	O
patients	NN	O	O
,	NN	O	O
17	NN	O	O
of	NN	O	O
whom	NN	O	O
were	NN	O	O
clinically	NN	O	O
suspected	NN	O	O
of	NN	O	O
having	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
eight	NN	O	O
cases	NN	O	O
with	NN	O	O
15q11	NN	O	O
-	NN	O	O
12	NN	O	O
deletion	NN	O	O
were	NN	O	O
very	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
originally	NN	O	O
described	NN	O	O
in	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
group	NN	O	O
of	NN	O	O
normal	NN	O	O
karyotype	NN	O	O
patients	NN	O	O
is	NN	O	O
heterogeneous	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
features	NN	O	O
do	NN	O	O
not	NN	O	O
strictly	NN	O	O
correspond	NN	O	O
to	NN	O	O
the	NN	O	O
clinical	NN	O	O
definition	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
PWS	NN	O	B-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
deletion	NN	O	O
of	NN	O	O
15q11	NN	O	O
-	NN	O	O
12	NN	O	O
can	NN	O	O
neither	NN	O	O
explain	NN	O	O
the	NN	O	O
apparently	NN	O	O
balanced	NN	O	O
translocations	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15	NN	O	O
nor	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
small	NN	O	O
supernumerary	NN	O	O
metacentric	NN	O	O
chromosomes	NN	O	O
corresponding	NN	O	O
to	NN	O	O
an	NN	O	O
isochromosome	NN	O	O
15	NN	O	O
for	NN	O	O
band	NN	O	O
15q11	NN	O	O
observed	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
North	NN	O	O
American	NN	O	O
Jewish	NN	O	O
populations	NN	O	O
:	NN	O	O
geographic	NN	O	O
variations	NN	O	O
and	NN	O	O
origin	NN	O	O
.	NN	O	O

From	NN	O	O
data	NN	O	O
collected	NN	O	O
in	NN	O	O
a	NN	O	O
North	NN	O	O
American	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
heterozygote	NN	O	O
screening	NN	O	O
program	NN	O	O
,	NN	O	O
the	NN	O	O
TSD	NN	O	B-Disease
carrier	NN	O	O
frequency	NN	O	O
among	NN	O	O
46	NN	O	O
,	NN	O	O
304	NN	O	O
Jewish	NN	O	O
individuals	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
.	NN	O	O

0324	NN	O	O
(	NN	O	O
1	NN	O	O
in	NN	O	O
31	NN	O	O
individuals	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
frequency	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
earlier	NN	O	O
estimates	NN	O	O
based	NN	O	O
on	NN	O	O
TSD	NN	O	B-Disease
incidence	NN	O	O
data	NN	O	O
.	NN	O	O

TSD	NN	O	B-Disease
carrier	NN	O	O
frequencies	NN	O	O
were	NN	O	O
then	NN	O	O
examined	NN	O	O
by	NN	O	O
single	NN	O	O
country	NN	O	O
and	NN	O	O
single	NN	O	O
region	NN	O	O
of	NN	O	O
origin	NN	O	O
in	NN	O	O
28	NN	O	O
,	NN	O	O
029	NN	O	O
Jews	NN	O	O
within	NN	O	O
this	NN	O	O
sample	NN	O	O
for	NN	O	O
whom	NN	O	O
such	NN	O	O
data	NN	O	O
were	NN	O	O
available	NN	O	O
for	NN	O	O
analysis	NN	O	O
.	NN	O	O

Jews	NN	O	O
with	NN	O	O
Polish	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
Russian	NN	O	O
ancestry	NN	O	O
constituted	NN	O	O
88	NN	O	O
%	NN	O	O
of	NN	O	O
this	NN	O	O
sample	NN	O	O
and	NN	O	O
had	NN	O	O
a	NN	O	O
TSD	NN	O	B-Disease
carrier	NN	O	O
frequency	NN	O	O
of	NN	O	O
.	NN	O	O

0327	NN	O	O
.	NN	O	O

No	NN	O	O
TSD	NN	O	B-Disease
carriers	NN	O	O
were	NN	O	O
observed	NN	O	O
among	NN	O	O
the	NN	O	O
166	NN	O	O
Jews	NN	O	O
of	NN	O	O
Near	NN	O	O
Eastern	NN	O	O
origins	NN	O	O
.	NN	O	O

Relative	NN	O	O
to	NN	O	O
Jews	NN	O	O
of	NN	O	O
Polish	NN	O	O
and	NN	O	O
Russian	NN	O	O
origins	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
twofold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
TSD	NN	O	B-Disease
carrier	NN	O	O
frequency	NN	O	O
in	NN	O	O
Jews	NN	O	O
of	NN	O	O
Austrian	NN	O	O
,	NN	O	O
Hungarian	NN	O	O
,	NN	O	O
and	NN	O	O
Czechoslovakian	NN	O	O
origins	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
.	NN	O	O
005	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
TSD	NN	O	B-Disease
gene	NN	O	O
proliferated	NN	O	O
among	NN	O	O
the	NN	O	O
antecedents	NN	O	O
of	NN	O	O
modern	NN	O	O
Ashkenazi	NN	O	O
Jewry	NN	O	O
after	NN	O	O
the	NN	O	O
Second	NN	O	O
Diaspora	NN	O	O
(	NN	O	O
70	NN	O	O
A	NN	O	O
.	NN	O	O
D	NN	O	O
.	NN	O	O
)	NN	O	O
and	NN	O	O
before	NN	O	O
their	NN	O	O
major	NN	O	O
migrations	NN	O	O
to	NN	O	O
regions	NN	O	O
of	NN	O	O
Poland	NN	O	O
and	NN	O	O
Russia	NN	O	O
(	NN	O	O
before	NN	O	O
1100	NN	O	O
A	NN	O	O
.	NN	O	O
D	NN	O	O
.	NN	O	O
)	NN	O	O
.	NN	O	O

Human	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
sixth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
and	NN	O	O
no	NN	O	O
haemostasis	NN	O	B-Disease
abnormality	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
case	NN	O	O
of	NN	O	O
human	NN	O	O
complete	NN	O	O
C6	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
reported	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
,	NN	O	O
a	NN	O	O
31	NN	O	O
year	NN	O	O
old	NN	O	O
white	NN	O	O
male	NN	O	O
,	NN	O	O
was	NN	O	O
seen	NN	O	O
on	NN	O	O
the	NN	O	O
occasion	NN	O	O
of	NN	O	O
an	NN	O	O
isolated	NN	O	O
episode	NN	O	O
of	NN	O	O
meningococcal	NN	O	B-Disease
meningitis	NN	O	I-Disease
.	NN	O	O

Serum	NN	O	O
complement	NN	O	O
hemolytic	NN	O	O
and	NN	O	O
bactericidal	NN	O	O
activities	NN	O	O
were	NN	O	O
lacking	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
restored	NN	O	O
to	NN	O	O
normal	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
appropriate	NN	O	O
amounts	NN	O	O
of	NN	O	O
purified	NN	O	O
C6	NN	O	O
.	NN	O	O

No	NN	O	O
hemostatic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
were	NN	O	O
observed	NN	O	O
.	NN	O	O
.	NN	O	O

Absence	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	B-Disease
meningococcemia	NN	O	I-Disease
presenting	NN	O	O
as	NN	O	O
vasculitis	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
previously	NN	O	O
healthy	NN	O	O
40	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
man	NN	O	O
presenting	NN	O	O
with	NN	O	O
fever	NN	O	B-Disease
,	NN	O	O
arthritis	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
cutaneous	NN	O	B-Disease
vasculitis	NN	O	I-Disease
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
chronic	NN	O	B-Disease
meningococcemia	NN	O	I-Disease
.	NN	O	O

Evaluation	NN	O	O
of	NN	O	O
his	NN	O	O
complement	NN	O	O
system	NN	O	O
showed	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
functional	NN	O	O
and	NN	O	O
antigenic	NN	O	O
C7	NN	O	O
,	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
complete	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
.	NN	O	O

Study	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
family	NN	O	O
spanning	NN	O	O
four	NN	O	O
generations	NN	O	O
showed	NN	O	O
heterozygous	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
C7	NN	O	I-Disease
in	NN	O	O
five	NN	O	O
members	NN	O	O
.	NN	O	O

Chronic	NN	O	B-Disease
neisserial	NN	O	I-Disease
infection	NN	O	I-Disease
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
must	NN	O	O
be	NN	O	O
distinguished	NN	O	O
from	NN	O	O
other	NN	O	O
causes	NN	O	O
of	NN	O	O
cutaneous	NN	O	B-Disease
vasculitis	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
discoid	NN	O	I-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
heterozygote	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
siblings	NN	O	O
with	NN	O	O
chronic	NN	O	B-Disease
discoid	NN	O	I-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
and	NN	O	O
several	NN	O	O
family	NN	O	O
members	NN	O	O
were	NN	O	O
found	NN	O	O
with	NN	O	O
heterozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

An	NN	O	O
association	NN	O	O
with	NN	O	O
histocompatibility	NN	O	O
markers	NN	O	O
HLA	NN	O	O
-	NN	O	O
B18	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
Dw2	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
slow	NN	O	O
allotype	NN	O	O
of	NN	O	O
factor	NN	O	O
B	NN	O	O
was	NN	O	O
present	NN	O	O
.	NN	O	O

Linkage	NN	O	O
studies	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
suggested	NN	O	O
a	NN	O	O
close	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
gene	NN	O	O
and	NN	O	O
genes	NN	O	O
coding	NN	O	O
for	NN	O	O
B18	NN	O	O
,	NN	O	O
Dw2	NN	O	O
,	NN	O	O
and	NN	O	O
BfS	NN	O	O
antigens	NN	O	O
.	NN	O	O

One	NN	O	O
HLA	NN	O	O
-	NN	O	O
ACB	NN	O	O
/	NN	O	O
DBf	NN	O	O
recombinant	NN	O	O
was	NN	O	O
observed	NN	O	O
showing	NN	O	O
closer	NN	O	O
linkage	NN	O	O
between	NN	O	O
HLA	NN	O	O
-	NN	O	O
D	NN	O	O
and	NN	O	O
Bf	NN	O	O
than	NN	O	O
between	NN	O	O
HLA	NN	O	O
-	NN	O	O
B	NN	O	O
and	NN	O	O
Bf	NN	O	O
.	NN	O	O
.	NN	O	O

Severe	NN	O	B-Disease
-	NN	O	I-Disease
glucose	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
chronic	NN	O	O
hemolytic	NN	O	B-Disease
anemia	NN	O	I-Disease
,	NN	O	O
granulocyte	NN	O	B-Disease
dysfunction	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
infections	NN	O	O
:	NN	O	O
description	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
molecular	NN	O	O
variant	NN	O	O
(	NN	O	O
G6PD	NN	O	O
Barcelona	NN	O	O
)	NN	O	O
.	NN	O	O

Molecular	NN	O	O
,	NN	O	O
kinetic	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
studies	NN	O	O
were	NN	O	O
carried	NN	O	O
out	NN	O	O
on	NN	O	O
erythrocytes	NN	O	O
and	NN	O	O
leukocytes	NN	O	O
in	NN	O	O
a	NN	O	O
Spanish	NN	O	O
male	NN	O	O
with	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
congenital	NN	O	B-Disease
nonspherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
(	NN	O	O
CNSHA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
infections	NN	O	O
.	NN	O	O

G6PD	NN	O	O
activity	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
patients	NN	O	O
red	NN	O	O
cells	NN	O	O
and	NN	O	O
was	NN	O	O
about	NN	O	O
2	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
in	NN	O	O
leukocytes	NN	O	O
.	NN	O	O

Molecular	NN	O	O
studies	NN	O	O
using	NN	O	O
standard	NN	O	O
methods	NN	O	O
(	NN	O	O
WHO	NN	O	O
,	NN	O	O
1967	NN	O	O
)	NN	O	O
showed	NN	O	O
G6PD	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
slightly	NN	O	O
fast	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
at	NN	O	O
pH	NN	O	O
8	NN	O	O
.	NN	O	O

0	NN	O	O
with	NN	O	O
otherwise	NN	O	O
normal	NN	O	O
properties	NN	O	O
(	NN	O	O
heat	NN	O	O
stability	NN	O	O
at	NN	O	O
46	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
apparent	NN	O	O
affinity	NN	O	O
for	NN	O	O
substrates	NN	O	O
,	NN	O	O
optimum	NN	O	O
pH	NN	O	O
,	NN	O	O
and	NN	O	O
utilization	NN	O	O
of	NN	O	O
substrate	NN	O	O
analogues	NN	O	O
)	NN	O	O
.	NN	O	O

Other	NN	O	O
tests	NN	O	O
showed	NN	O	O
the	NN	O	O
patients	NN	O	O
granulocytes	NN	O	O
to	NN	O	O
engulf	NN	O	O
latex	NN	O	O
particles	NN	O	O
normally	NN	O	O
,	NN	O	O
but	NN	O	O
to	NN	O	O
have	NN	O	O
impaired	NN	O	O
reduction	NN	O	O
of	NN	O	O
nitroblue	NN	O	O
tetrazolium	NN	O	O
and	NN	O	O
ferricytochrome	NN	O	O
-	NN	O	O
c	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
reduced	NN	O	O
iodination	NN	O	O
.	NN	O	O

Chemotaxis	NN	O	O
and	NN	O	O
random	NN	O	O
migration	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
granulocytes	NN	O	O
were	NN	O	O
normal	NN	O	O
as	NN	O	O
were	NN	O	O
myeloperoxidase	NN	O	O
,	NN	O	O
leukocyte	NN	O	O
alkaline	NN	O	O
phosphatase	NN	O	O
(	NN	O	O
LAP	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
ultrastructural	NN	O	O
features	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
characteristics	NN	O	O
of	NN	O	O
G6PD	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
differed	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
all	NN	O	O
previously	NN	O	O
reported	NN	O	O
variants	NN	O	O
associated	NN	O	O
with	NN	O	O
CNSHA	NN	O	B-Disease
,	NN	O	O
so	NN	O	O
the	NN	O	O
present	NN	O	O
variant	NN	O	O
was	NN	O	O
provisionally	NN	O	O
called	NN	O	O
G6PD	NN	O	O
Barcelona	NN	O	O
to	NN	O	O
distinguish	NN	O	O
it	NN	O	O
from	NN	O	O
other	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
previously	NN	O	O
described	NN	O	O
.	NN	O	O

Possible	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
the	NN	O	O
severe	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
G6PD	NN	O	I-Disease
in	NN	O	O
erythrocytes	NN	O	O
and	NN	O	O
granulocytes	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
immunologic	NN	O	O
specific	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
enzyme	NN	O	O
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
.	NN	O	O

The	NN	O	O
description	NN	O	O
of	NN	O	O
13	NN	O	O
new	NN	O	O
variants	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
362	NN	O	O
males	NN	O	O
from	NN	O	O
various	NN	O	O
regions	NN	O	O
of	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
were	NN	O	O
screened	NN	O	O
for	NN	O	O
red	NN	O	O
cell	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
activity	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
six	NN	O	O
G6PD	NN	O	B-Disease
deficient	NN	O	I-Disease
individuals	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
characterization	NN	O	O
of	NN	O	O
G6PD	NN	O	O
purified	NN	O	O
from	NN	O	O
these	NN	O	O
subjects	NN	O	O
has	NN	O	O
revealed	NN	O	O
13	NN	O	O
new	NN	O	O
variants	NN	O	O
and	NN	O	O
several	NN	O	O
copies	NN	O	O
of	NN	O	O
previously	NN	O	O
described	NN	O	O
forms	NN	O	O
of	NN	O	O
G6PD	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
illustrates	NN	O	O
the	NN	O	O
extreme	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
among	NN	O	O
the	NN	O	O
people	NN	O	O
of	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
.	NN	O	O
.	NN	O	O

Heterogeneity	NN	O	O
of	NN	O	O
"	NN	O	O
Mediterranean	NN	O	O
type	NN	O	O
"	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Spain	NN	O	O
and	NN	O	O
description	NN	O	O
of	NN	O	O
two	NN	O	O
new	NN	O	O
variants	NN	O	O
associated	NN	O	O
with	NN	O	O
favism	NN	O	B-Disease
.	NN	O	O

Glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
;	NN	O	O
EC	NN	O	O
1	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

49	NN	O	O
from	NN	O	O
thirty	NN	O	O
-	NN	O	O
six	NN	O	O
unrelated	NN	O	O
Spanish	NN	O	O
males	NN	O	O
was	NN	O	O
partially	NN	O	O
purified	NN	O	O
from	NN	O	O
blood	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
variants	NN	O	O
were	NN	O	O
characterized	NN	O	O
biochemically	NN	O	O
and	NN	O	O
electrophoretically	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
methods	NN	O	O
recommended	NN	O	O
by	NN	O	O
the	NN	O	O
world	NN	O	O
Health	NN	O	O
Organization	NN	O	O
.	NN	O	O

Subjects	NN	O	O
were	NN	O	O
from	NN	O	O
multiple	NN	O	O
geographic	NN	O	O
regions	NN	O	O
within	NN	O	O
Spain	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
suffered	NN	O	O
from	NN	O	O
hemolytic	NN	O	B-Disease
anemia	NN	O	I-Disease
,	NN	O	O
either	NN	O	O
acute	NN	O	O
(	NN	O	O
34	NN	O	O
cases	NN	O	O
)	NN	O	O
or	NN	O	O
chronic	NN	O	O
nonspherocytic	NN	O	O
(	NN	O	O
2	NN	O	O
cases	NN	O	O
)	NN	O	O
.	NN	O	O

Almost	NN	O	O
all	NN	O	O
the	NN	O	O
variants	NN	O	O
studied	NN	O	O
presented	NN	O	O
residual	NN	O	O
erythrocyte	NN	O	O
G6PD	NN	O	O
activity	NN	O	O
ranging	NN	O	O
from	NN	O	O
0	NN	O	O
to	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
,	NN	O	O
and	NN	O	O
five	NN	O	O
different	NN	O	O
mutants	NN	O	O
were	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
deficient	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Three	NN	O	O
variants	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
others	NN	O	O
previously	NN	O	O
described	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
(	NN	O	O
11	NN	O	O
cases	NN	O	O
)	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Athens	NN	O	O
-	NN	O	O
like	NN	O	O
(	NN	O	O
3	NN	O	O
cases	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Union	NN	O	O
(	NN	O	O
2	NN	O	O
cases	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
variants	NN	O	O
were	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
numerous	NN	O	O
variants	NN	O	O
already	NN	O	O
reported	NN	O	O
and	NN	O	O
have	NN	O	O
been	NN	O	O
considered	NN	O	O
as	NN	O	O
new	NN	O	O
mutants	NN	O	O
.	NN	O	O

Provisionally	NN	O	O
they	NN	O	O
are	NN	O	O
called	NN	O	O
G6PD	NN	O	O
Betica	NN	O	O
(	NN	O	O
19	NN	O	O
cases	NN	O	O
)	NN	O	O
and	NN	O	O
G6PD	NN	O	O
Menorca	NN	O	O
(	NN	O	O
1	NN	O	O
case	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
constitutes	NN	O	O
the	NN	O	O
first	NN	O	O
attempt	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
deficient	NN	O	B-Disease
G6PD	NN	O	I-Disease
variants	NN	O	O
found	NN	O	O
in	NN	O	O
Spain	NN	O	O
and	NN	O	O
supplies	NN	O	O
new	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
molecular	NN	O	O
characteristics	NN	O	O
of	NN	O	O
deficient	NN	O	O
variants	NN	O	O
and	NN	O	O
their	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
important	NN	O	O
findings	NN	O	O
can	NN	O	O
be	NN	O	O
summarized	NN	O	O
as	NN	O	O
follows	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
The	NN	O	O
Spanish	NN	O	O
population	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
an	NN	O	O
important	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

(	NN	O	O
2	NN	O	O
)	NN	O	O
Although	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
is	NN	O	O
very	NN	O	O
frequent	NN	O	O
,	NN	O	O
it	NN	O	O
presents	NN	O	O
a	NN	O	O
relatively	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

(	NN	O	O
3	NN	O	O
)	NN	O	O
Favism	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
observed	NN	O	O
associated	NN	O	O
with	NN	O	O
all	NN	O	O
kinds	NN	O	O
of	NN	O	O
variants	NN	O	O
described	NN	O	O
here	NN	O	O
.	NN	O	O

(	NN	O	O
4	NN	O	O
)	NN	O	O
G6PD	NN	O	O
Betica	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
variant	NN	O	O
found	NN	O	O
in	NN	O	O
subjects	NN	O	O
of	NN	O	O
Southern	NN	O	O
Spanish	NN	O	O
origin	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
associated	NN	O	O
with	NN	O	O
favism	NN	O	B-Disease
in	NN	O	O
all	NN	O	O
cases	NN	O	O
except	NN	O	O
one	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variant	NN	O	O
(	NN	O	O
G6PD	NN	O	O
Nagano	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
new	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
variant	NN	O	O
associated	NN	O	O
with	NN	O	O
chronic	NN	O	B-Disease
nonspherocytic	NN	O	I-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
was	NN	O	O
reported	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
,	NN	O	O
a	NN	O	O
6	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
Japanese	NN	O	O
male	NN	O	O
,	NN	O	O
was	NN	O	O
noticed	NN	O	O
to	NN	O	O
have	NN	O	O
hemolytic	NN	O	B-Disease
anemia	NN	O	I-Disease
soon	NN	O	O
after	NN	O	O
birth	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
made	NN	O	O
at	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
2	NN	O	O
.	NN	O	O

He	NN	O	O
had	NN	O	O
episodes	NN	O	O
of	NN	O	O
hemolytic	NN	O	B-Disease
crisis	NN	O	I-Disease
several	NN	O	O
times	NN	O	O
after	NN	O	O
upper	NN	O	B-Disease
respiratory	NN	O	I-Disease
infection	NN	O	I-Disease
.	NN	O	O

G6PD	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
was	NN	O	O
5	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
enzymatic	NN	O	O
characteristics	NN	O	O
were	NN	O	O
examined	NN	O	O
when	NN	O	O
he	NN	O	O
was	NN	O	O
5	NN	O	O
years	NN	O	O
old	NN	O	O
,	NN	O	O
and	NN	O	O
his	NN	O	O
G6PD	NN	O	O
showed	NN	O	O
faster	NN	O	O
-	NN	O	O
than	NN	O	O
-	NN	O	O
normal	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
,	NN	O	O
low	NN	O	O
Km	NN	O	O
G6P	NN	O	O
,	NN	O	O
high	NN	O	O
Km	NN	O	O
NADP	NN	O	O
,	NN	O	O
low	NN	O	O
Ki	NN	O	O
NADPH	NN	O	O
,	NN	O	O
normal	NN	O	O
utilization	NN	O	O
of	NN	O	O
substrate	NN	O	O
analogues	NN	O	O
,	NN	O	O
heat	NN	O	O
instability	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
normal	NN	O	O
pH	NN	O	O
optimum	NN	O	O
curve	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
this	NN	O	O
was	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
new	NN	O	O
variant	NN	O	O
and	NN	O	O
was	NN	O	O
designated	NN	O	O
G6PD	NN	O	O
Nagano	NN	O	O
.	NN	O	O

Infection	NN	O	O
-	NN	O	O
induced	NN	O	O
hemolysis	NN	O	B-Disease
and	NN	O	O
chronic	NN	O	B-Disease
hemolytic	NN	O	I-Disease
anemia	NN	O	I-Disease
seem	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
markedly	NN	O	O
impaired	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
and	NN	O	O
thermal	NN	O	O
instability	NN	O	O
.	NN	O	O

Heterozygous	NN	O	O
C2	NN	O	B-Disease
-	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
myasthenia	NN	O	B-Disease
gravis	NN	O	I-Disease
.	NN	O	O

Complement	NN	O	B-Disease
deficiency	NN	O	I-Disease
states	NN	O	O
in	NN	O	O
myasthenia	NN	O	B-Disease
gravis	NN	O	I-Disease
(	NN	O	O
MG	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
19	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
woman	NN	O	O
with	NN	O	O
typical	NN	O	O
MG	NN	O	B-Disease
and	NN	O	O
heterozygous	NN	O	O
C2	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
along	NN	O	O
with	NN	O	O
HLA	NN	O	O
typing	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
her	NN	O	O
immediate	NN	O	O
family	NN	O	O
.	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
:	NN	O	O
increased	NN	O	O
plasma	NN	O	O
content	NN	O	O
of	NN	O	O
saturated	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
.	NN	O	O

With	NN	O	O
a	NN	O	O
new	NN	O	O
method	NN	O	O
we	NN	O	O
measured	NN	O	O
the	NN	O	O
saturated	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
of	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
hemizygotes	NN	O	O
,	NN	O	O
ALD	NN	O	B-Disease
heterozygotes	NN	O	O
,	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

ALD	NN	O	B-Disease
hemizygotes	NN	O	O
showed	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
hexacosanoate	NN	O	O
(	NN	O	O
C26	NN	O	O
fatty	NN	O	O
acid	NN	O	O
)	NN	O	O
which	NN	O	O
represented	NN	O	O
0	NN	O	O
.	NN	O	O

081	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

0066	NN	O	O
%	NN	O	O
(	NN	O	O
SEM	NN	O	O
)	NN	O	O
of	NN	O	O
total	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
0	NN	O	O
.	NN	O	O

015	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

0032	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
controls	NN	O	O
.	NN	O	O

C25	NN	O	O
,	NN	O	O
C24	NN	O	O
,	NN	O	O
and	NN	O	O
C23	NN	O	O
fatty	NN	O	O
acids	NN	O	O
were	NN	O	O
also	NN	O	O
increased	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
C22	NN	O	O
and	NN	O	O
C20	NN	O	O
fatty	NN	O	O
acids	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

C26	NN	O	O
levels	NN	O	O
were	NN	O	O
also	NN	O	O
increased	NN	O	O
in	NN	O	O
most	NN	O	O
ALD	NN	O	B-Disease
heterozygotes	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
mean	NN	O	O
level	NN	O	O
0	NN	O	O
.	NN	O	O

057	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

0063	NN	O	O
%	NN	O	O
of	NN	O	O
total	NN	O	O
fatty	NN	O	O
acids	NN	O	O
.	NN	O	O

The	NN	O	O
technique	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
carrier	NN	O	O
identification	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
evaluation	NN	O	O
of	NN	O	O
therapy	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
high	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
in	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
lipoprotein	NN	O	O
profiles	NN	O	O
and	NN	O	O
high	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
were	NN	O	O
characterized	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
disease	NN	O	I-Disease
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Abnormalities	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
HDL	NN	O	I-Disease
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
their	NN	O	O
increased	NN	O	O
atherogenesis	NN	O	O
and	NN	O	O
excessive	NN	O	O
deposits	NN	O	O
of	NN	O	O
tissue	NN	O	O
sterols	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
low	NN	O	O
or	NN	O	O
low	NN	O	O
-	NN	O	O
normal	NN	O	O
concentrations	NN	O	O
of	NN	O	O
plasma	NN	O	O
cholesterol	NN	O	O
(	NN	O	O
165	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
25	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
)	NN	O	O
and	NN	O	O
low	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
(	NN	O	O
LDL	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
HDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
CTX	NN	O	B-Disease
plasmas	NN	O	O
was	NN	O	O
14	NN	O	O
.	NN	O	O

5	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
3	NN	O	O
.	NN	O	O

2	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
,	NN	O	O
about	NN	O	O
one	NN	O	O
-	NN	O	O
third	NN	O	O
the	NN	O	O
normal	NN	O	O
value	NN	O	O
.	NN	O	O

The	NN	O	O
low	NN	O	O
HDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
reflects	NN	O	O
a	NN	O	O
low	NN	O	O
concentration	NN	O	O
and	NN	O	O
an	NN	O	O
abnormal	NN	O	O
lipid	NN	O	O
composition	NN	O	O
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
HDL	NN	O	O
.	NN	O	O

Relative	NN	O	O
to	NN	O	O
normal	NN	O	O
HDL	NN	O	O
,	NN	O	O
the	NN	O	O
cholesteryl	NN	O	O
esters	NN	O	O
are	NN	O	O
low	NN	O	O
,	NN	O	O
free	NN	O	O
cholesterol	NN	O	O
and	NN	O	O
phospholipids	NN	O	O
essentially	NN	O	O
normal	NN	O	O
,	NN	O	O
and	NN	O	O
triglycerides	NN	O	O
increased	NN	O	O
.	NN	O	O

The	NN	O	O
ratio	NN	O	O
of	NN	O	O
apoprotein	NN	O	O
(	NN	O	O
apo	NN	O	O
)	NN	O	O
to	NN	O	O
total	NN	O	O
cholesterol	NN	O	O
in	NN	O	O
the	NN	O	O
HDL	NN	O	O
of	NN	O	O
CTX	NN	O	B-Disease
was	NN	O	O
two	NN	O	O
to	NN	O	O
three	NN	O	O
times	NN	O	O
greater	NN	O	O
than	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
CTX	NN	O	B-Disease
HDL	NN	O	O
,	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
apoAI	NN	O	O
to	NN	O	O
apoAII	NN	O	O
was	NN	O	O
high	NN	O	O
,	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
apoC	NN	O	O
low	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
normally	NN	O	O
minor	NN	O	O
form	NN	O	O
of	NN	O	O
apoAI	NN	O	O
increased	NN	O	O
relative	NN	O	O
to	NN	O	O
other	NN	O	O
forms	NN	O	O
.	NN	O	O

The	NN	O	O
HDL	NN	O	O
in	NN	O	O
electron	NN	O	O
micrographs	NN	O	O
appeared	NN	O	O
normal	NN	O	O
morphologically	NN	O	O
and	NN	O	O
in	NN	O	O
particle	NN	O	O
size	NN	O	O
.	NN	O	O

The	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
lipoprotein	NN	O	O
distribution	NN	O	O
profile	NN	O	O
and	NN	O	O
composition	NN	O	O
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
HDL	NN	O	O
result	NN	O	O
from	NN	O	O
metabolic	NN	O	O
defects	NN	O	O
that	NN	O	O
are	NN	O	O
not	NN	O	O
understood	NN	O	O
but	NN	O	O
may	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
in	NN	O	O
bile	NN	O	O
acid	NN	O	O
synthesis	NN	O	O
in	NN	O	O
CTX	NN	O	B-Disease
.	NN	O	O

As	NN	O	O
a	NN	O	O
consequence	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
probable	NN	O	O
that	NN	O	O
the	NN	O	O
normal	NN	O	O
functions	NN	O	O
of	NN	O	O
the	NN	O	O
HDL	NN	O	O
,	NN	O	O
possibly	NN	O	O
including	NN	O	O
modulation	NN	O	O
of	NN	O	O
LDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
uptake	NN	O	O
and	NN	O	O
the	NN	O	O
removal	NN	O	O
of	NN	O	O
excess	NN	O	O
cholesterol	NN	O	O
from	NN	O	O
peripheral	NN	O	O
tissues	NN	O	O
,	NN	O	O
are	NN	O	O
perturbed	NN	O	O
significantly	NN	O	O
in	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

Genetics	NN	O	O
of	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
:	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
trait	NN	O	O
with	NN	O	O
high	NN	O	O
gene	NN	O	O
frequency	NN	O	O
in	NN	O	O
Sephardim	NN	O	O
of	NN	O	O
Moroccan	NN	O	O
origin	NN	O	O
.	NN	O	O

We	NN	O	O
described	NN	O	O
6	NN	O	O
patients	NN	O	O
(	NN	O	O
from	NN	O	O
3	NN	O	O
families	NN	O	O
)	NN	O	O
affected	NN	O	O
with	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

All	NN	O	O
are	NN	O	O
Sephardic	NN	O	O
Jews	NN	O	O
of	NN	O	O
Moroccan	NN	O	O
extraction	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
CTX	NN	O	B-Disease
patients	NN	O	O
diagnosed	NN	O	O
in	NN	O	O
the	NN	O	O
world	NN	O	O
(	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
50	NN	O	O
including	NN	O	O
our	NN	O	O
6	NN	O	O
patients	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
are	NN	O	O
probably	NN	O	O
dealing	NN	O	O
with	NN	O	O
an	NN	O	O
ethnic	NN	O	O
subgroup	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
CTX	NN	O	O
gene	NN	O	O
frequency	NN	O	O
,	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
1	NN	O	O
/	NN	O	O
108	NN	O	O
.	NN	O	O

Since	NN	O	O
there	NN	O	O
are	NN	O	O
differences	NN	O	O
in	NN	O	O
expression	NN	O	O
in	NN	O	O
this	NN	O	O
disease	NN	O	O
,	NN	O	O
we	NN	O	O
recommend	NN	O	O
cholestanol	NN	O	O
study	NN	O	O
in	NN	O	O
cases	NN	O	O
of	NN	O	O
undiagnosed	NN	O	O
cataract	NN	O	B-Disease
or	NN	O	O
tendinous	NN	O	B-Disease
xanthomas	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
or	NN	O	O
early	NN	O	O
adolescence	NN	O	O
.	NN	O	O

The	NN	O	O
diagnosis	NN	O	O
in	NN	O	O
CTX	NN	O	B-Disease
is	NN	O	O
important	NN	O	O
not	NN	O	O
only	NN	O	O
for	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
veiw	NN	O	O
of	NN	O	O
possible	NN	O	O
treatment	NN	O	O
.	NN	O	O
.	NN	O	O

New	NN	O	O
genetic	NN	O	O
variants	NN	O	O
of	NN	O	O
glucose	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
in	NN	O	O
Italy	NN	O	O
.	NN	O	O

Six	NN	O	O
new	NN	O	O
variants	NN	O	O
of	NN	O	O
human	NN	O	O
erythrocyte	NN	O	O
G6PD	NN	O	O
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
them	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
Italian	NN	O	O
males	NN	O	O
and	NN	O	O
all	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
only	NN	O	O
two	NN	O	O
with	NN	O	O
signs	NN	O	O
of	NN	O	O
haemolysis	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
and	NN	O	O
other	NN	O	O
variants	NN	O	O
reported	NN	O	O
in	NN	O	O
the	NN	O	O
literature	NN	O	O
,	NN	O	O
which	NN	O	O
must	NN	O	O
thus	NN	O	O
far	NN	O	O
be	NN	O	O
regarded	NN	O	O
as	NN	O	O
sporadic	NN	O	O
,	NN	O	O
are	NN	O	O
found	NN	O	O
to	NN	O	O
map	NN	O	O
in	NN	O	O
parts	NN	O	O
of	NN	O	O
Italy	NN	O	O
where	NN	O	O
common	NN	O	O
types	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
are	NN	O	O
also	NN	O	O
prevalent	NN	O	O
.	NN	O	O
.	NN	O	O

Variants	NN	O	O
of	NN	O	O
erythrocyte	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
in	NN	O	O
Bulgarian	NN	O	O
populations	NN	O	O
.	NN	O	O

Ten	NN	O	O
variants	NN	O	O
of	NN	O	O
erythrocyte	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
22	NN	O	O
patients	NN	O	O
with	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
from	NN	O	O
three	NN	O	O
districts	NN	O	O
of	NN	O	O
Bulgaria	NN	O	O
.	NN	O	O

Corinth	NN	O	O
-	NN	O	O
like	NN	O	O
and	NN	O	O
Fayoum	NN	O	O
-	NN	O	O
like	NN	O	O
variants	NN	O	O
were	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
;	NN	O	O
Mediterranean	NN	O	O
,	NN	O	O
Ohut	NN	O	O
II	NN	O	O
,	NN	O	O
Kilgore	NN	O	O
,	NN	O	O
Boston	NN	O	O
,	NN	O	O
Poznan	NN	O	O
,	NN	O	O
and	NN	O	O
Panay	NN	O	O
variants	NN	O	O
and	NN	O	O
two	NN	O	O
new	NN	O	O
variants	NN	O	O
,	NN	O	O
Petrich	NN	O	O
and	NN	O	O
Gotze	NN	O	O
Delchev	NN	O	O
,	NN	O	O
were	NN	O	O
each	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
or	NN	O	O
two	NN	O	O
carriers	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
revealed	NN	O	O
between	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
polymorphism	NN	O	O
.	NN	O	O
.	NN	O	O

Nephropathy	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Nephropathy	NN	O	B-Disease
was	NN	O	O
detected	NN	O	O
in	NN	O	O
five	NN	O	O
of	NN	O	O
32	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
who	NN	O	O
were	NN	O	O
participating	NN	O	O
in	NN	O	O
a	NN	O	O
study	NN	O	O
of	NN	O	O
transfer	NN	O	O
factor	NN	O	O
(	NN	O	O
TF	NN	O	O
)	NN	O	O
therapy	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
patients	NN	O	O
,	NN	O	O
nephropathy	NN	O	B-Disease
was	NN	O	O
present	NN	O	O
before	NN	O	O
TF	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
appear	NN	O	O
changed	NN	O	O
by	NN	O	O
TF	NN	O	O
therapy	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
subsequently	NN	O	O
developed	NN	O	O
progressive	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
requiring	NN	O	O
dialysis	NN	O	O
beginning	NN	O	O
5	NN	O	O
1	NN	O	O
/	NN	O	O
2	NN	O	O
years	NN	O	O
after	NN	O	O
TF	NN	O	O
therapy	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
patients	NN	O	O
,	NN	O	O
decreased	NN	O	O
renal	NN	O	O
function	NN	O	O
appeared	NN	O	O
very	NN	O	O
soon	NN	O	O
after	NN	O	O
the	NN	O	O
administration	NN	O	O
of	NN	O	O
TF	NN	O	O
.	NN	O	O

One	NN	O	O
patient	NN	O	O
showed	NN	O	O
gradually	NN	O	O
decreasing	NN	O	O
renal	NN	O	O
function	NN	O	O
beginning	NN	O	O
after	NN	O	O
two	NN	O	O
years	NN	O	O
of	NN	O	O
TF	NN	O	O
therapy	NN	O	O
.	NN	O	O

An	NN	O	O
additional	NN	O	O
patient	NN	O	O
was	NN	O	O
identified	NN	O	O
who	NN	O	O
died	NN	O	O
with	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
without	NN	O	O
having	NN	O	O
received	NN	O	O
TF	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
renal	NN	O	B-Disease
failure	NN	O	I-Disease
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
more	NN	O	O
frequently	NN	O	O
than	NN	O	O
generally	NN	O	O
recognized	NN	O	O
and	NN	O	O
that	NN	O	O
administration	NN	O	O
of	NN	O	O
TF	NN	O	O
may	NN	O	O
precipitate	NN	O	O
or	NN	O	O
accelerate	NN	O	O
the	NN	O	O
renal	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
patients	NN	O	O
with	NN	O	O
this	NN	O	O
syndrome	NN	O	O
.	NN	O	O
.	NN	O	O

Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
:	NN	O	O
cellular	NN	O	O
impairments	NN	O	O
and	NN	O	O
their	NN	O	O
implication	NN	O	O
for	NN	O	O
carrier	NN	O	O
detection	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
two	NN	O	O
male	NN	O	O
siblings	NN	O	O
were	NN	O	O
affected	NN	O	O
with	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
studied	NN	O	O
using	NN	O	O
G	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
PD	NN	O	O
isoenzymes	NN	O	O
as	NN	O	O
an	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
marker	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
cellular	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

Isolated	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
cell	NN	O	O
types	NN	O	O
from	NN	O	O
the	NN	O	O
doubly	NN	O	O
heterozygous	NN	O	O
mother	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
males	NN	O	O
seemingly	NN	O	O
failed	NN	O	O
to	NN	O	O
express	NN	O	O
the	NN	O	O
G	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
PD	NN	O	O
allele	NN	O	O
in	NN	O	O
cis	NN	O	O
position	NN	O	O
with	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
allele	NN	O	O
while	NN	O	O
her	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
expressed	NN	O	O
both	NN	O	O
G	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
PD	NN	O	O
alleles	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
a	NN	O	O
histogram	NN	O	O
analysis	NN	O	O
of	NN	O	O
platelet	NN	O	O
size	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
population	NN	O	O
of	NN	O	O
abnormally	NN	O	O
small	NN	O	O
platelets	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
propositus	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
mother	NN	O	O
had	NN	O	O
no	NN	O	O
appreciable	NN	O	O
small	NN	O	O
platelet	NN	O	O
subpopulation	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
culture	NN	O	O
of	NN	O	O
hemopoietic	NN	O	O
progenitor	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
mother	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
progenitor	NN	O	O
cells	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
allele	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
G	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
PD	NN	O	O
type	NN	O	O
linked	NN	O	O
with	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
allele	NN	O	O
were	NN	O	O
detected	NN	O	O
.	NN	O	O

The	NN	O	O
proportion	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
progenitors	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
among	NN	O	O
more	NN	O	O
primitive	NN	O	O
progenitors	NN	O	O
(	NN	O	O
those	NN	O	O
giving	NN	O	O
rise	NN	O	O
to	NN	O	O
later	NN	O	O
appearing	NN	O	O
colonies	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
suggests	NN	O	O
that	NN	O	O
selection	NN	O	O
against	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
defect	NN	O	I-Disease
takes	NN	O	O
place	NN	O	O
in	NN	O	O
the	NN	O	O
hemopoietic	NN	O	O
system	NN	O	O
of	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
female	NN	O	O
and	NN	O	O
offers	NN	O	O
a	NN	O	O
possible	NN	O	O
means	NN	O	O
of	NN	O	O
carrier	NN	O	O
detection	NN	O	O
in	NN	O	O
some	NN	O	O
women	NN	O	O
.	NN	O	O

Linkage	NN	O	O
studies	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
revealed	NN	O	O
one	NN	O	O
example	NN	O	O
of	NN	O	O
probable	NN	O	O
recombination	NN	O	O
between	NN	O	O
the	NN	O	O
loci	NN	O	O
for	NN	O	O
WAS	NN	O	B-Disease
and	NN	O	O
G	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
PD	NN	O	O
among	NN	O	O
three	NN	O	O
informative	NN	O	O
subjects	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
two	NN	O	O
loci	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
closely	NN	O	O
linked	NN	O	O
on	NN	O	O
the	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
CT	NN	O	O
pattern	NN	O	O
in	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
new	NN	O	O
CT	NN	O	O
pattern	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
2	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
,	NN	O	O
which	NN	O	O
the	NN	O	O
authors	NN	O	O
call	NN	O	O
Type	NN	O	O
II	NN	O	O
,	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
posterior	NN	O	O
periventricular	NN	O	O
areas	NN	O	O
of	NN	O	O
decreased	NN	O	O
attenuation	NN	O	O
around	NN	O	O
the	NN	O	O
trigone	NN	O	O
on	NN	O	O
non	NN	O	O
-	NN	O	O
contrast	NN	O	O
scans	NN	O	O
after	NN	O	O
contrast	NN	O	O
infusion	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
striking	NN	O	O
enhancement	NN	O	O
of	NN	O	O
various	NN	O	O
white	NN	O	O
-	NN	O	O
matter	NN	O	O
structures	NN	O	O
(	NN	O	O
tracts	NN	O	O
or	NN	O	O
fiber	NN	O	O
systems	NN	O	O
)	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
internal	NN	O	O
capsules	NN	O	O
,	NN	O	O
corpus	NN	O	O
callosum	NN	O	O
,	NN	O	O
corona	NN	O	O
radiata	NN	O	O
,	NN	O	O
forceps	NN	O	O
major	NN	O	O
,	NN	O	O
and	NN	O	O
cerebral	NN	O	O
peduncles	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
different	NN	O	O
from	NN	O	O
numerous	NN	O	O
previous	NN	O	O
descriptions	NN	O	O
of	NN	O	O
the	NN	O	O
CT	NN	O	O
pattern	NN	O	O
in	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

Type	NN	O	B-Disease
II	NN	O	I-Disease
ALD	NN	O	I-Disease
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
have	NN	O	O
been	NN	O	O
seen	NN	O	O
in	NN	O	O
any	NN	O	O
other	NN	O	O
leukoencephalopathy	NN	O	B-Disease
and	NN	O	O
is	NN	O	O
probably	NN	O	O
specific	NN	O	O
for	NN	O	O
a	NN	O	O
phenotypic	NN	O	O
variant	NN	O	O
or	NN	O	O
an	NN	O	O
evolving	NN	O	O
stage	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Further	NN	O	O
evidence	NN	O	O
for	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
.	NN	O	O

Four	NN	O	O
new	NN	O	O
G6PD	NN	O	O
variants	NN	O	O
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
in	NN	O	O
individuals	NN	O	O
from	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
the	NN	O	O
previously	NN	O	O
reported	NN	O	O
Markham	NN	O	O
variant	NN	O	O
and	NN	O	O
the	NN	O	O
newly	NN	O	O
characterized	NN	O	O
Salata	NN	O	O
variant	NN	O	O
may	NN	O	O
be	NN	O	O
widely	NN	O	O
distributed	NN	O	O
in	NN	O	O
Papua	NN	O	O
New	NN	O	O
Guinea	NN	O	O
.	NN	O	O

Th	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
together	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
previously	NN	O	O
published	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
degree	NN	O	O
of	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
that	NN	O	O
is	NN	O	O
much	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
other	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
world	NN	O	O
where	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
common	NN	O	O
.	NN	O	O
.	NN	O	O

Increased	NN	O	O
incidence	NN	O	O
of	NN	O	O
cataracts	NN	O	B-Disease
in	NN	O	O
male	NN	O	O
subjects	NN	O	O
deficient	NN	O	B-Disease
in	NN	O	I-Disease
glucose	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
RBCs	NN	O	O
was	NN	O	O
found	NN	O	O
significantly	NN	O	O
more	NN	O	O
frequently	NN	O	O
in	NN	O	O
210	NN	O	O
male	NN	O	O
cataractous	NN	O	B-Disease
patients	NN	O	O
than	NN	O	O
in	NN	O	O
672	NN	O	O
control	NN	O	O
subjects	NN	O	O
of	NN	O	O
Sardinian	NN	O	O
origin	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
deficiency	NN	O	O
was	NN	O	O
increasingly	NN	O	O
higher	NN	O	O
in	NN	O	O
presenile	NN	O	O
cataracts	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
group	NN	O	O
,	NN	O	O
the	NN	O	O
incidence	NN	O	O
of	NN	O	O
cortical	NN	O	B-Disease
and	NN	O	I-Disease
total	NN	O	I-Disease
cataracts	NN	O	I-Disease
was	NN	O	O
also	NN	O	O
increased	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
decrease	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
lens	NN	O	O
,	NN	O	O
which	NN	O	O
accompanies	NN	O	O
its	NN	O	O
deficiency	NN	O	O
in	NN	O	O
the	NN	O	O
erythrocyte	NN	O	O
,	NN	O	O
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cataracto	NN	O	O
-	NN	O	O
genesis	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
should	NN	O	O
be	NN	O	O
added	NN	O	O
to	NN	O	O
other	NN	O	O
conditions	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
galactosemic	NN	O	B-Disease
states	NN	O	O
and	NN	O	O
riboflavin	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
where	NN	O	O
cataracts	NN	O	B-Disease
represent	NN	O	O
a	NN	O	O
sensitive	NN	O	O
indicator	NN	O	O
of	NN	O	O
metabolic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
of	NN	O	O
the	NN	O	O
RBC	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
human	NN	O	O
mitochondrial	NN	O	O
very	NN	O	B-Disease
-	NN	O	I-Disease
long	NN	O	I-Disease
-	NN	O	I-Disease
chain	NN	O	I-Disease
acyl	NN	O	I-Disease
-	NN	O	I-Disease
CoA	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
deficiency	NN	O	I-Disease
causing	NN	O	O
cardiomyopathy	NN	O	B-Disease
and	NN	O	O
sudden	NN	O	B-Disease
death	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
.	NN	O	O

beta	NN	O	O
-	NN	O	O
Oxidation	NN	O	O
of	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
provides	NN	O	O
the	NN	O	O
major	NN	O	O
source	NN	O	O
of	NN	O	O
energy	NN	O	O
in	NN	O	O
the	NN	O	O
heart	NN	O	O
.	NN	O	O

Defects	NN	O	O
in	NN	O	O
enzymes	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
pathway	NN	O	O
cause	NN	O	O
sudden	NN	O	B-Disease
,	NN	O	I-Disease
unexplained	NN	O	I-Disease
death	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
,	NN	O	O
acute	NN	O	O
hepatic	NN	O	B-Disease
encephalopathy	NN	O	I-Disease
or	NN	O	O
liver	NN	O	B-Disease
failure	NN	O	I-Disease
,	NN	O	O
skeletal	NN	O	B-Disease
myopathy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
cardiomyopathy	NN	O	B-Disease
.	NN	O	O

Very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
dehydrogenase	NN	O	O
[	NN	O	O
VLCAD	NN	O	O
;	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
-	NN	O	O
acyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
(	NN	O	O
acceptor	NN	O	O
)	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
-	NN	O	O
oxidoreductase	NN	O	O
,	NN	O	O
EC	NN	O	O
1	NN	O	O
.	NN	O	O
3	NN	O	O
3	NN	O	O
.	NN	O	O
99	NN	O	O
99	NN	O	O
.	NN	O	O
13	NN	O	O
]	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
first	NN	O	O
step	NN	O	O
in	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
the	NN	O	O
human	NN	O	O
VLCAD	NN	O	O
cDNA	NN	O	O
and	NN	O	O
gene	NN	O	O
and	NN	O	O
determined	NN	O	O
the	NN	O	O
complete	NN	O	O
nucleotide	NN	O	O
sequences	NN	O	O
.	NN	O	O

Polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
amplification	NN	O	O
of	NN	O	O
VLCAD	NN	O	O
mRNA	NN	O	O
and	NN	O	O
genomic	NN	O	O
exons	NN	O	O
defined	NN	O	O
the	NN	O	O
molecular	NN	O	O
defects	NN	O	O
in	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
who	NN	O	O
presented	NN	O	O
with	NN	O	O
unexplained	NN	O	O
cardiac	NN	O	B-Disease
arrest	NN	O	I-Disease
and	NN	O	O
cardiomyopathy	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
one	NN	O	O
,	NN	O	O
a	NN	O	O
homozygous	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
consensus	NN	O	O
dinucleotide	NN	O	O
of	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
(	NN	O	O
g	NN	O	O
+	NN	O	O
1	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
a	NN	O	O
)	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
universal	NN	O	O
skipping	NN	O	O
of	NN	O	O
the	NN	O	O
prior	NN	O	O
exon	NN	O	O
(	NN	O	O
exon	NN	O	O
11	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
patient	NN	O	O
was	NN	O	O
a	NN	O	O
compound	NN	O	O
heterozygote	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
,	NN	O	O
C1837	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
,	NN	O	O
changing	NN	O	O
the	NN	O	O
arginine	NN	O	O
at	NN	O	O
residue	NN	O	O
613	NN	O	O
to	NN	O	O
tryptophan	NN	O	O
on	NN	O	O
one	NN	O	O
allele	NN	O	O
and	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
deletion	NN	O	O
at	NN	O	O
the	NN	O	O
intron	NN	O	O
-	NN	O	O
exon	NN	O	O
6	NN	O	O
boundary	NN	O	O
as	NN	O	O
the	NN	O	O
second	NN	O	O
mutation	NN	O	O
.	NN	O	O

This	NN	O	O
initial	NN	O	O
delineation	NN	O	O
of	NN	O	O
human	NN	O	O
mutations	NN	O	O
in	NN	O	O
VLCAD	NN	O	O
suggests	NN	O	O
that	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
reduces	NN	O	O
myocardial	NN	O	O
fatty	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
and	NN	O	O
energy	NN	O	O
production	NN	O	O
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
cardiomyopathy	NN	O	B-Disease
and	NN	O	O
sudden	NN	O	B-Disease
death	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
.	NN	O	O

Aberrant	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
product	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
220	NN	O	O
-	NN	O	O
kilodalton	NN	O	O
nuclear	NN	O	O
phosphoprotein	NN	O	O
in	NN	O	O
normal	NN	O	O
cells	NN	O	O
,	NN	O	O
including	NN	O	O
breast	NN	O	O
ductal	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
18	NN	O	O
of	NN	O	O
20	NN	O	O
tumor	NN	O	B-Disease
cell	NN	O	O
lines	NN	O	O
derived	NN	O	O
from	NN	O	O
tissues	NN	O	O
other	NN	O	O
than	NN	O	O
breast	NN	O	O
and	NN	O	O
ovary	NN	O	O
.	NN	O	O

In	NN	O	O
16	NN	O	O
of	NN	O	O
17	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
lines	NN	O	O
and	NN	O	O
17	NN	O	O
of	NN	O	O
17	NN	O	O
samples	NN	O	O
of	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
malignant	NN	O	O
effusions	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
localized	NN	O	O
mainly	NN	O	O
in	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Absence	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
or	NN	O	O
aberrant	NN	O	O
subcellular	NN	O	O
location	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
to	NN	O	O
a	NN	O	O
variable	NN	O	O
extent	NN	O	O
in	NN	O	O
histological	NN	O	O
sections	NN	O	O
of	NN	O	O
many	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
biopsies	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
BRCA1	NN	O	B-Disease
abnormalities	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
many	NN	O	O
breast	NN	O	B-Disease
cancers	NN	O	I-Disease
,	NN	O	O
sporadic	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
familial	NN	O	O
.	NN	O	O
.	NN	O	O

Mapping	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
8	NN	O	O
.	NN	O	O

ATP7B	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
altered	NN	O	O
in	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
WD	NN	O	B-Disease
)	NN	O	O
patients	NN	O	O
,	NN	O	O
lies	NN	O	O
in	NN	O	O
a	NN	O	O
block	NN	O	O
of	NN	O	O
homology	NN	O	O
shared	NN	O	O
between	NN	O	O
human	NN	O	O
chromosome	NN	O	O
13q14	NN	O	O
and	NN	O	O
the	NN	O	O
central	NN	O	O
region	NN	O	O
of	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
14	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
mapped	NN	O	O
the	NN	O	O
murine	NN	O	O
homologue	NN	O	O
of	NN	O	O
ATP7B	NN	O	O
(	NN	O	O
Atp7b	NN	O	O
)	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
8	NN	O	O
by	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrid	NN	O	O
analysis	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
80	NN	O	O
interspecific	NN	O	O
backcross	NN	O	O
offspring	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
position	NN	O	O
Atp7b	NN	O	O
close	NN	O	O
to	NN	O	O
D8Mit3	NN	O	O
and	NN	O	O
another	NN	O	O
ATPase	NN	O	O
locus	NN	O	O
,	NN	O	O
Atp4b	NN	O	O
,	NN	O	O
on	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
8	NN	O	O
.	NN	O	O

ATP4B	NN	O	O
lies	NN	O	O
in	NN	O	O
13q34	NN	O	O
and	NN	O	O
is	NN	O	O
separated	NN	O	O
from	NN	O	O
ATP7B	NN	O	O
by	NN	O	O
several	NN	O	O
loci	NN	O	O
whose	NN	O	O
mouse	NN	O	O
homologues	NN	O	O
map	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
14	NN	O	O
.	NN	O	O

The	NN	O	O
assignment	NN	O	O
of	NN	O	O
Atp7b	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
8	NN	O	O
identifies	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
region	NN	O	O
of	NN	O	O
homology	NN	O	O
between	NN	O	O
this	NN	O	O
chromosome	NN	O	O
and	NN	O	O
human	NN	O	O
chromosome	NN	O	O
13	NN	O	O
.	NN	O	O

This	NN	O	O
assignment	NN	O	O
suggests	NN	O	O
a	NN	O	O
possible	NN	O	O
location	NN	O	O
for	NN	O	O
the	NN	O	O
toxic	NN	O	O
milk	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
as	NN	O	O
a	NN	O	O
homologue	NN	O	O
of	NN	O	O
WD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
in	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlation	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
,	NN	O	O
discovered	NN	O	O
in	NN	O	O
1994	NN	O	O
,	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
80	NN	O	O
-	NN	O	O
90	NN	O	O
%	NN	O	O
lifetime	NN	O	O
risk	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
analysed	NN	O	O
60	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
for	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Twenty	NN	O	O
-	NN	O	O
two	NN	O	O
different	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
32	NN	O	O
families	NN	O	O
(	NN	O	O
53	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
14	NN	O	O
are	NN	O	O
previously	NN	O	O
unreported	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
to	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
incidence	NN	O	O
within	NN	O	O
each	NN	O	O
family	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
transition	NN	O	O
in	NN	O	O
risk	NN	O	O
such	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
third	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
lower	NN	O	O
proportion	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Haplotype	NN	O	O
analysis	NN	O	O
supports	NN	O	O
previous	NN	O	O
data	NN	O	O
which	NN	O	O
suggest	NN	O	O
some	NN	O	O
BRCA1	NN	O	O
mutation	NN	O	O
carriers	NN	O	O
have	NN	O	O
common	NN	O	O
ancestors	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
examples	NN	O	O
where	NN	O	O
recurrent	NN	O	O
mutations	NN	O	O
appear	NN	O	O
to	NN	O	O
have	NN	O	O
arisen	NN	O	O
independently	NN	O	O
.	NN	O	O
.	NN	O	O

Hereditary	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
and	NN	O	O
recurrent	NN	O	O
meningococcal	NN	O	B-Disease
infection	NN	O	I-Disease
:	NN	O	O
investigations	NN	O	O
of	NN	O	O
an	NN	O	O
Irish	NN	O	O
family	NN	O	O
using	NN	O	O
a	NN	O	O
novel	NN	O	O
haemolytic	NN	O	O
screening	NN	O	O
assay	NN	O	O
for	NN	O	O
complement	NN	O	O
activity	NN	O	O
and	NN	O	O
C7	NN	O	O
M	NN	O	O
/	NN	O	O
N	NN	O	O
allotyping	NN	O	O
.	NN	O	O

Terminal	NN	O	B-Disease
complement	NN	O	I-Disease
component	NN	O	I-Disease
deficiency	NN	O	I-Disease
predisposes	NN	O	O
to	NN	O	O
meningococcal	NN	O	B-Disease
infection	NN	O	I-Disease
and	NN	O	O
is	NN	O	O
inherited	NN	O	O
in	NN	O	O
an	NN	O	O
autosomal	NN	O	O
co	NN	O	O
-	NN	O	O
dominant	NN	O	O
manner	NN	O	O
.	NN	O	O

An	NN	O	O
Irish	NN	O	O
family	NN	O	O
is	NN	O	O
described	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
2	NN	O	O
of	NN	O	O
3	NN	O	O
brothers	NN	O	O
had	NN	O	O
recurrent	NN	O	O
meningococcal	NN	O	B-Disease
infection	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
novel	NN	O	O
screening	NN	O	O
assay	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
investigate	NN	O	O
for	NN	O	O
terminal	NN	O	B-Disease
complement	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
2	NN	O	O
affected	NN	O	O
brothers	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
completely	NN	O	B-Disease
deficient	NN	O	I-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
seventh	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C7	NN	O	O
)	NN	O	O
.	NN	O	O

Enzyme	NN	O	O
-	NN	O	O
linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
for	NN	O	O
C7	NN	O	O
revealed	NN	O	O
lower	NN	O	O
than	NN	O	O
normal	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
brother	NN	O	O
and	NN	O	O
parents	NN	O	O
.	NN	O	O

C7	NN	O	O
M	NN	O	O
/	NN	O	O
N	NN	O	O
protein	NN	O	O
polymorphism	NN	O	O
allotyping	NN	O	O
,	NN	O	O
used	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
segregation	NN	O	O
of	NN	O	O
the	NN	O	O
C7	NN	O	B-Disease
deficiency	NN	O	I-Disease
genes	NN	O	O
,	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
apparently	NN	O	O
complement	NN	O	O
sufficient	NN	O	O
brother	NN	O	O
was	NN	O	O
heterozygous	NN	O	O
C7	NN	O	B-Disease
deficient	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
carrier	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
deficiency	NN	O	O
genes	NN	O	O
.	NN	O	O

Complement	NN	O	O
screening	NN	O	O
should	NN	O	O
be	NN	O	O
carried	NN	O	O
out	NN	O	O
in	NN	O	O
any	NN	O	O
individual	NN	O	O
suffering	NN	O	O
recurrent	NN	O	O
meningococcal	NN	O	B-Disease
infection	NN	O	I-Disease
or	NN	O	O
infection	NN	O	O
with	NN	O	O
an	NN	O	O
uncommon	NN	O	O
meningococcal	NN	O	B-Disease
serogroup	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
complement	NN	O	B-Disease
deficient	NN	O	I-Disease
patients	NN	O	O
allows	NN	O	O
the	NN	O	O
implementation	NN	O	O
of	NN	O	O
strategies	NN	O	O
to	NN	O	O
prevent	NN	O	O
recurrent	NN	O	O
infection	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
subtotal	NN	O	B-Disease
complement	NN	O	I-Disease
C6	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminally	NN	O	O
truncated	NN	O	O
but	NN	O	O
functionally	NN	O	O
active	NN	O	O
C6	NN	O	O
.	NN	O	O

Individuals	NN	O	O
with	NN	O	O
subtotal	NN	O	B-Disease
complement	NN	O	I-Disease
C6	NN	O	I-Disease
deficiency	NN	O	I-Disease
possess	NN	O	O
a	NN	O	O
C6	NN	O	O
molecule	NN	O	O
that	NN	O	O
is	NN	O	O
14	NN	O	O
%	NN	O	O
shorter	NN	O	O
than	NN	O	O
normal	NN	O	O
C6	NN	O	O
and	NN	O	O
present	NN	O	O
in	NN	O	O
low	NN	O	O
but	NN	O	O
detectable	NN	O	O
concentrations	NN	O	O
(	NN	O	O
1	NN	O	O
-	NN	O	O
2	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
mean	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
dysmorphic	NN	O	O
C6	NN	O	O
is	NN	O	O
bactericidally	NN	O	O
active	NN	O	O
and	NN	O	O
lacks	NN	O	O
an	NN	O	O
epitope	NN	O	O
that	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
most	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminal	NN	O	O
part	NN	O	O
of	NN	O	O
C6	NN	O	O
using	NN	O	O
C6	NN	O	O
cDNA	NN	O	O
fragments	NN	O	O
expressed	NN	O	O
as	NN	O	O
fusion	NN	O	O
proteins	NN	O	O
in	NN	O	O
the	NN	O	O
pUEX	NN	O	O
expression	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
thus	NN	O	O
predicted	NN	O	O
that	NN	O	O
the	NN	O	O
abnormal	NN	O	O
C6	NN	O	O
molecule	NN	O	O
might	NN	O	O
be	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminally	NN	O	O
truncated	NN	O	O
and	NN	O	O
sought	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
an	NN	O	O
area	NN	O	O
approximately	NN	O	O
14	NN	O	O
%	NN	O	O
from	NN	O	O
the	NN	O	O
carboxy	NN	O	O
-	NN	O	O
terminal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
.	NN	O	O

By	NN	O	O
sequencing	NN	O	O
PCR	NN	O	O
-	NN	O	O
amplified	NN	O	O
products	NN	O	O
from	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
,	NN	O	O
in	NN	O	O
three	NN	O	O
individuals	NN	O	O
from	NN	O	O
two	NN	O	O
families	NN	O	O
,	NN	O	O
a	NN	O	O
mutation	NN	O	O
that	NN	O	O
might	NN	O	O
plausibly	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
defect	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
have	NN	O	O
an	NN	O	O
abnormal	NN	O	O
5	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
15	NN	O	O
,	NN	O	O
which	NN	O	O
would	NN	O	O
probably	NN	O	O
prevent	NN	O	O
splicing	NN	O	O
.	NN	O	O

An	NN	O	O
in	NN	O	O
-	NN	O	O
frame	NN	O	O
stop	NN	O	O
codon	NN	O	O
is	NN	O	O
found	NN	O	O
17	NN	O	O
codons	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
intron	NN	O	O
boundary	NN	O	O
,	NN	O	O
which	NN	O	O
would	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
truncated	NN	O	O
polypeptide	NN	O	O
13	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
smaller	NN	O	O
than	NN	O	O
normal	NN	O	O
C6	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
was	NN	O	O
unexpected	NN	O	O
,	NN	O	O
as	NN	O	O
earlier	NN	O	O
studies	NN	O	O
mapped	NN	O	O
the	NN	O	O
C5b	NN	O	O
binding	NN	O	O
site	NN	O	O
,	NN	O	O
or	NN	O	O
a	NN	O	O
putative	NN	O	O
enzymatic	NN	O	O
region	NN	O	O
,	NN	O	O
to	NN	O	O
this	NN	O	O
part	NN	O	O
of	NN	O	O
C6	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
all	NN	O	O
three	NN	O	O
subjects	NN	O	O
were	NN	O	O
probably	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
both	NN	O	O
subtotal	NN	O	B-Disease
C6	NN	O	I-Disease
and	NN	O	I-Disease
complete	NN	O	I-Disease
C6	NN	O	I-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
possible	NN	O	O
dominant	NN	O	O
-	NN	O	O
negative	NN	O	O
RNA	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
trinucleotide	NN	O	O
expansion	NN	O	O
mutation	NN	O	O
causing	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
is	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
expanded	NN	O	O
repeat	NN	O	O
causes	NN	O	O
the	NN	O	O
clinically	NN	O	O
variable	NN	O	O
and	NN	O	O
multisystemic	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
are	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
particularly	NN	O	O
difficult	NN	O	O
to	NN	O	O
rationalize	NN	O	O
the	NN	O	O
dominant	NN	O	O
inheritance	NN	O	O
with	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
the	NN	O	O
expansion	NN	O	O
mutation	NN	O	O
lies	NN	O	O
outside	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
-	NN	O	O
encoding	NN	O	O
gene	NN	O	O
elements	NN	O	O
,	NN	O	O
and	NN	O	O
should	NN	O	O
not	NN	O	O
be	NN	O	O
translated	NN	O	O
into	NN	O	O
protein	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
use	NN	O	O
muscle	NN	O	O
biopsies	NN	O	O
from	NN	O	O
classical	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
patients	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
transcripts	NN	O	O
from	NN	O	O
both	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
expanded	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
genes	NN	O	O
in	NN	O	O
patient	NN	O	O
muscle	NN	O	O
,	NN	O	O
and	NN	O	O
compare	NN	O	O
the	NN	O	O
results	NN	O	O
to	NN	O	O
normal	NN	O	O
and	NN	O	O
myopathic	NN	O	B-Disease
controls	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
relatively	NN	O	O
small	NN	O	O
decreases	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
RNA	NN	O	O
in	NN	O	O
the	NN	O	O
total	NN	O	O
RNA	NN	O	O
pool	NN	O	O
from	NN	O	O
muscle	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
these	NN	O	O
reductions	NN	O	O
were	NN	O	O
not	NN	O	O
disease	NN	O	O
specific	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
poly	NN	O	O
(	NN	O	O
A	NN	O	O
)	NN	O	O
+	NN	O	O
RNA	NN	O	O
showed	NN	O	O
dramatic	NN	O	O
decreases	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
mutant	NN	O	O
and	NN	O	O
normal	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
RNAs	NN	O	O
,	NN	O	O
and	NN	O	O
these	NN	O	O
changes	NN	O	O
were	NN	O	O
disease	NN	O	O
-	NN	O	O
specific	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
molecular	NN	O	O
pathogenetic	NN	O	O
mechanism	NN	O	O
for	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
both	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
expanded	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
genes	NN	O	O
are	NN	O	O
transcribed	NN	O	O
in	NN	O	O
patient	NN	O	O
muscle	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
abnormal	NN	O	O
expansion	NN	O	O
-	NN	O	O
containing	NN	O	O
RNA	NN	O	O
has	NN	O	O
a	NN	O	O
dominant	NN	O	O
effect	NN	O	O
on	NN	O	O
RNA	NN	O	O
metabolism	NN	O	O
by	NN	O	O
preventing	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
poly	NN	O	O
(	NN	O	O
A	NN	O	O
)	NN	O	O
+	NN	O	O
RNA	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
expansion	NN	O	O
mutation	NN	O	O
to	NN	O	O
alter	NN	O	O
accumulation	NN	O	O
of	NN	O	O
poly	NN	O	O
(	NN	O	O
A	NN	O	O
)	NN	O	O
+	NN	O	O
RNA	NN	O	O
in	NN	O	O
trans	NN	O	O
suggests	NN	O	O
that	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
-	NN	O	O
negative	NN	O	O
mutation	NN	O	O
manifested	NN	O	O
at	NN	O	O
the	NN	O	O
RNA	NN	O	O
level	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
strong	NN	O	O
candidate	NN	O	O
for	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
gene	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

A	NN	O	O
strong	NN	O	O
candidate	NN	O	O
for	NN	O	O
the	NN	O	O
17q	NN	O	O
-	NN	O	O
linked	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
influences	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
by	NN	O	O
positional	NN	O	O
cloning	NN	O	O
methods	NN	O	O
.	NN	O	O

Probable	NN	O	O
predisposing	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
five	NN	O	O
of	NN	O	O
eight	NN	O	O
kindreds	NN	O	O
presumed	NN	O	O
to	NN	O	O
segregate	NN	O	O
BRCA1	NN	O	O
susceptibility	NN	O	O
alleles	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
include	NN	O	O
an	NN	O	O
11	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
,	NN	O	O
a	NN	O	O
1	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
insertion	NN	O	O
,	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
,	NN	O	O
a	NN	O	O
missense	NN	O	O
substitution	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
inferred	NN	O	O
regulatory	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
numerous	NN	O	O
tissues	NN	O	O
,	NN	O	O
including	NN	O	O
breast	NN	O	O
and	NN	O	O
ovary	NN	O	O
,	NN	O	O
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
predicted	NN	O	O
protein	NN	O	O
of	NN	O	O
1863	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

This	NN	O	O
protein	NN	O	O
contains	NN	O	O
a	NN	O	O
zinc	NN	O	O
finger	NN	O	O
domain	NN	O	O
in	NN	O	O
its	NN	O	O
amino	NN	O	O
-	NN	O	O
terminal	NN	O	O
region	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
otherwise	NN	O	O
unrelated	NN	O	O
to	NN	O	O
previously	NN	O	O
described	NN	O	O
proteins	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
should	NN	O	O
facilitate	NN	O	O
early	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
in	NN	O	O
some	NN	O	O
individuals	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
biology	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
galactosemia	NN	O	B-Disease
(	NN	O	O
type	NN	O	O
1	NN	O	O
)	NN	O	O
mutations	NN	O	O
in	NN	O	O
Japanese	NN	O	O
.	NN	O	O

We	NN	O	O
characterized	NN	O	O
two	NN	O	O
novel	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyltransferase	NN	O	O
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
gene	NN	O	O
in	NN	O	O
two	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
with	NN	O	O
GALT	NN	O	B-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
identified	NN	O	O
N314D	NN	O	O
and	NN	O	O
R333W	NN	O	O
mutations	NN	O	O
,	NN	O	O
previously	NN	O	O
found	NN	O	O
in	NN	O	O
Caucasians	NN	O	O
.	NN	O	O

One	NN	O	O
novel	NN	O	O
missense	NN	O	O
mutation	NN	O	O
was	NN	O	O
an	NN	O	O
G	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
transition	NN	O	O
in	NN	O	O
exon	NN	O	O
8	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
arginine	NN	O	O
by	NN	O	O
histidine	NN	O	O
at	NN	O	O
the	NN	O	O
codon	NN	O	O
231	NN	O	O
(	NN	O	O
R231H	NN	O	O
)	NN	O	O
.	NN	O	O

GALT	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
R231H	NN	O	O
mutant	NN	O	O
construct	NN	O	O
was	NN	O	O
reduced	NN	O	O
to	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
controls	NN	O	O
in	NN	O	O
a	NN	O	O
COS	NN	O	O
cell	NN	O	O
expression	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
was	NN	O	O
a	NN	O	O
splicing	NN	O	O
mutation	NN	O	O
,	NN	O	O
an	NN	O	O
A	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
G	NN	O	O
transition	NN	O	O
at	NN	O	O
the	NN	O	O
38th	NN	O	O
nucleotide	NN	O	O
in	NN	O	O
exon	NN	O	O
3	NN	O	O
(	NN	O	O
318A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
38	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
GALT	NN	O	O
cDNA	NN	O	O
by	NN	O	O
activating	NN	O	O
a	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
acceptor	NN	O	O
site	NN	O	O
.	NN	O	O

In	NN	O	O
seven	NN	O	O
Japanese	NN	O	O
families	NN	O	O
(	NN	O	O
14	NN	O	O
alleles	NN	O	O
for	NN	O	O
classic	NN	O	O
form	NN	O	O
and	NN	O	O
one	NN	O	O
allele	NN	O	O
for	NN	O	O
Duarte	NN	O	O
variant	NN	O	O
)	NN	O	O
with	NN	O	O
GALT	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
R231H	NN	O	O
and	NN	O	O
318A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
only	NN	O	O
on	NN	O	O
both	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
proband	NN	O	O
.	NN	O	O

The	NN	O	O
N314D	NN	O	O
and	NN	O	O
R333W	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
on	NN	O	O
one	NN	O	O
allele	NN	O	O
each	NN	O	O
.	NN	O	O

The	NN	O	O
Q188R	NN	O	O
was	NN	O	O
prevalent	NN	O	O
in	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
Japanese	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
N314D	NN	O	O
mutation	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
Duarte	NN	O	O
variant	NN	O	O
in	NN	O	O
Japanese	NN	O	O
persons	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
.	NN	O	O

We	NN	O	O
speculate	NN	O	O
that	NN	O	O
classic	NN	O	B-Disease
galactosemia	NN	O	I-Disease
mutations	NN	O	O
appear	NN	O	O
to	NN	O	O
differ	NN	O	O
between	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
patients	NN	O	O
.	NN	O	O

Our	NN	O	O
limited	NN	O	O
data	NN	O	O
set	NN	O	O
on	NN	O	O
galactosemia	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
Japanese	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
N314D	NN	O	O
GALT	NN	O	O
mutation	NN	O	O
encoding	NN	O	O
the	NN	O	O
Duarte	NN	O	O
variant	NN	O	O
arose	NN	O	O
before	NN	O	O
Asian	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
people	NN	O	O
diverged	NN	O	O
and	NN	O	O
that	NN	O	O
classic	NN	O	B-Disease
galactosemia	NN	O	I-Disease
mutations	NN	O	O
arose	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
accumulated	NN	O	O
after	NN	O	O
the	NN	O	O
divergence	NN	O	O
of	NN	O	O
Asian	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
populations	NN	O	O
.	NN	O	O
.	NN	O	O

Three	NN	O	O
novel	NN	O	O
aniridia	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
.	NN	O	O

Aniridia	NN	O	B-Disease
(	NN	O	O
iris	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
congenital	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
eye	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
aniridia	NN	O	B-Disease
(	NN	O	O
PAX6	NN	O	O
)	NN	O	O
gene	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
many	NN	O	O
patients	NN	O	O
from	NN	O	O
various	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
study	NN	O	O
reported	NN	O	O
here	NN	O	O
we	NN	O	O
describe	NN	O	O
PAX6	NN	O	O
mutations	NN	O	O
in	NN	O	O
one	NN	O	O
sporadic	NN	O	O
and	NN	O	O
five	NN	O	O
familial	NN	O	O
cases	NN	O	O
with	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O

Of	NN	O	O
the	NN	O	O
four	NN	O	O
different	NN	O	O
mutations	NN	O	O
identified	NN	O	O
,	NN	O	O
one	NN	O	O
was	NN	O	O
identical	NN	O	O
to	NN	O	O
a	NN	O	O
previously	NN	O	O
reported	NN	O	O
mutation	NN	O	O
(	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
at	NN	O	O
codon	NN	O	O
240	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
three	NN	O	O
were	NN	O	O
novel	NN	O	O
two	NN	O	O
in	NN	O	O
the	NN	O	O
glycine	NN	O	O
-	NN	O	O
rich	NN	O	O
region	NN	O	O
and	NN	O	O
one	NN	O	O
in	NN	O	O
the	NN	O	O
proline	NN	O	O
/	NN	O	O
serine	NN	O	O
/	NN	O	O
threonine	NN	O	O
-	NN	O	O
rich	NN	O	O
(	NN	O	O
PST	NN	O	O
)	NN	O	O
region	NN	O	O
.	NN	O	O

One	NN	O	O
PAX6	NN	O	O
mutation	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
PST	NN	O	O
region	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
cataracts	NN	O	B-Disease
in	NN	O	O
an	NN	O	O
aniridia	NN	O	B-Disease
family	NN	O	O
.	NN	O	O

Another	NN	O	O
splice	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
PST	NN	O	O
domain	NN	O	O
occurred	NN	O	O
in	NN	O	O
an	NN	O	O
aniridia	NN	O	B-Disease
patient	NN	O	O
with	NN	O	O
anosmia	NN	O	B-Disease
(	NN	O	O
inability	NN	O	O
to	NN	O	O
smell	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
six	NN	O	O
new	NN	O	O
aniridia	NN	O	B-Disease
cases	NN	O	O
reported	NN	O	O
here	NN	O	O
have	NN	O	O
mutations	NN	O	O
predicted	NN	O	O
to	NN	O	O
generate	NN	O	O
incomplete	NN	O	O
PAX6	NN	O	O
proteins	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
theory	NN	O	O
that	NN	O	O
human	NN	O	O
aniridia	NN	O	B-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
haploinsufficiency	NN	O	B-Disease
of	NN	O	I-Disease
PAX6	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

The	NN	O	O
carrier	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
185delAG	NN	O	O
mutation	NN	O	O
is	NN	O	O
approximately	NN	O	O
1	NN	O	O
percent	NN	O	O
in	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
individuals	NN	O	O
.	NN	O	O

Since	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
the	NN	O	O
first	NN	O	O
major	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
inherited	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
was	NN	O	O
cloned	NN	O	O
,	NN	O	O
more	NN	O	O
than	NN	O	O
50	NN	O	O
unique	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
germline	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
high	NN	O	O
-	NN	O	O
risk	NN	O	O
pedigrees	NN	O	O
,	NN	O	O
female	NN	O	O
carriers	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
have	NN	O	O
an	NN	O	O
80	NN	O	O
-	NN	O	O
90	NN	O	O
%	NN	O	O
lifetime	NN	O	O
risk	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
a	NN	O	O
40	NN	O	O
-	NN	O	O
50	NN	O	O
%	NN	O	O
risk	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mutation	NN	O	O
stats	NN	O	O
of	NN	O	O
individuals	NN	O	O
unselected	NN	O	O
for	NN	O	O
breast	NN	O	B-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
has	NN	O	O
not	NN	O	O
been	NN	O	O
determined	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
whether	NN	O	O
mutations	NN	O	O
in	NN	O	O
such	NN	O	O
individuals	NN	O	O
confer	NN	O	O
the	NN	O	O
same	NN	O	O
risk	NN	O	O
of	NN	O	O
cancer	NN	O	B-Disease
as	NN	O	O
in	NN	O	O
individuals	NN	O	O
from	NN	O	O
the	NN	O	O
high	NN	O	O
-	NN	O	O
risk	NN	O	O
families	NN	O	O
studied	NN	O	O
so	NN	O	O
far	NN	O	O
.	NN	O	O

Following	NN	O	O
the	NN	O	O
finding	NN	O	O
of	NN	O	O
a	NN	O	O
185delAG	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
in	NN	O	O
several	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
breast	NN	O	O
/	NN	O	O
ovarian	NN	O	O
families	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
in	NN	O	O
858	NN	O	O
Ashkenazim	NN	O	O
seeking	NN	O	O
genetic	NN	O	O
testing	NN	O	O
for	NN	O	O
conditions	NN	O	O
unrelated	NN	O	O
to	NN	O	O
cancer	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
in	NN	O	O
815	NN	O	O
reference	NN	O	O
individuals	NN	O	O
not	NN	O	O
selected	NN	O	O
for	NN	O	O
ethnic	NN	O	O
origin	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
the	NN	O	O
185delAG	NN	O	O
mutation	NN	O	O
in	NN	O	O
0	NN	O	O
.	NN	O	O

9	NN	O	O
%	NN	O	O
of	NN	O	O
Ashkenazim	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
limit	NN	O	O
,	NN	O	O
0	NN	O	O
.	NN	O	O
4	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
reference	NN	O	O
samples	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
one	NN	O	O
in	NN	O	O
a	NN	O	O
hundred	NN	O	O
women	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
descent	NN	O	O
may	NN	O	O
be	NN	O	O
at	NN	O	O
especially	NN	O	O
high	NN	O	O
risk	NN	O	O
of	NN	O	O
developing	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Identification	NN	O	O
and	NN	O	O
localization	NN	O	O
of	NN	O	O
huntingtin	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
human	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
lines	NN	O	O
with	NN	O	O
anti	NN	O	O
-	NN	O	O
fusion	NN	O	O
protein	NN	O	O
antibodies	NN	O	O
.	NN	O	O

The	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
phenotype	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
IT15	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
348	NN	O	O
-	NN	O	O
kDa	NN	O	O
protein	NN	O	O
named	NN	O	O
huntington	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
polyclonal	NN	O	O
and	NN	O	O
monoclonal	NN	O	O
anti	NN	O	O
-	NN	O	O
fusion	NN	O	O
protein	NN	O	O
antibodies	NN	O	O
to	NN	O	O
identify	NN	O	O
native	NN	O	O
huntingtin	NN	O	O
in	NN	O	O
rat	NN	O	O
,	NN	O	O
monkey	NN	O	O
,	NN	O	O
and	NN	O	O
human	NN	O	O
.	NN	O	O

Western	NN	O	O
blots	NN	O	O
revealed	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
the	NN	O	O
expected	NN	O	O
molecular	NN	O	O
weight	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
soluble	NN	O	O
fraction	NN	O	O
of	NN	O	O
rat	NN	O	O
and	NN	O	O
monkey	NN	O	O
brain	NN	O	O
tissues	NN	O	O
and	NN	O	O
lymphoblastoid	NN	O	O
cells	NN	O	O
from	NN	O	O
control	NN	O	O
cases	NN	O	O
.	NN	O	O

In	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
lines	NN	O	O
from	NN	O	O
juvenile	NN	O	O
-	NN	O	O
onset	NN	O	O
heterozygote	NN	O	O
HD	NN	O	B-Disease
cases	NN	O	O
,	NN	O	O
both	NN	O	O
normal	NN	O	O
and	NN	O	O
mutant	NN	O	O
huntingtin	NN	O	O
are	NN	O	O
expressed	NN	O	O
,	NN	O	O
and	NN	O	O
increasing	NN	O	O
repeat	NN	O	O
expansion	NN	O	O
leads	NN	O	O
to	NN	O	O
lower	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
protein	NN	O	O
.	NN	O	O

Immunocytochemistry	NN	O	O
indicates	NN	O	O
that	NN	O	O
huntingtin	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
neurons	NN	O	O
throughout	NN	O	O
the	NN	O	O
brain	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
highest	NN	O	O
levels	NN	O	O
evident	NN	O	O
in	NN	O	O
larger	NN	O	O
neurons	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
human	NN	O	O
striatum	NN	O	O
,	NN	O	O
huntingtin	NN	O	O
is	NN	O	O
enriched	NN	O	O
in	NN	O	O
a	NN	O	O
patch	NN	O	O
-	NN	O	O
like	NN	O	O
distribution	NN	O	O
,	NN	O	O
potentially	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
first	NN	O	O
areas	NN	O	O
affected	NN	O	O
in	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

Subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
huntingtin	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
cytosolic	NN	O	O
protein	NN	O	O
primarily	NN	O	O
found	NN	O	O
in	NN	O	O
somatodendritic	NN	O	O
regions	NN	O	O
.	NN	O	O

Huntingtin	NN	O	O
appears	NN	O	O
to	NN	O	O
particularly	NN	O	O
associate	NN	O	O
with	NN	O	O
microtubules	NN	O	O
,	NN	O	O
although	NN	O	O
some	NN	O	O
is	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
synaptic	NN	O	O
vesicles	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
localization	NN	O	O
of	NN	O	O
huntingtin	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
microtubules	NN	O	O
,	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
impairs	NN	O	O
the	NN	O	O
cytoskeletal	NN	O	O
anchoring	NN	O	O
or	NN	O	O
transport	NN	O	O
of	NN	O	O
mitochondria	NN	O	O
,	NN	O	O
vesicles	NN	O	O
,	NN	O	O
or	NN	O	O
other	NN	O	O
organelles	NN	O	O
or	NN	O	O
molecules	NN	O	O
.	NN	O	O
.	NN	O	O

Marked	NN	O	O
phenotypic	NN	O	O
heterogeneity	NN	O	O
associated	NN	O	O
with	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
sequence	NN	O	O
at	NN	O	O
the	NN	O	O
spinocerebellar	NN	O	B-Disease
ataxia	NN	O	I-Disease
3	NN	O	I-Disease
/	NN	O	I-Disease
Machado	NN	O	I-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
locus	NN	O	O
.	NN	O	O

The	NN	O	O
spinocerebellar	NN	O	B-Disease
ataxia	NN	O	I-Disease
3	NN	O	I-Disease
locus	NN	O	O
(	NN	O	O
SCA3	NN	O	B-Disease
)	NN	O	O
for	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
autosomal	NN	O	I-Disease
dominant	NN	O	I-Disease
cerebellar	NN	O	I-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
ADCA	NN	O	B-Disease
type	NN	O	I-Disease
I	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
clinically	NN	O	O
and	NN	O	O
genetically	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
neurodegenerative	NN	O	B-Disease
disorders	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
14q32	NN	O	O
.	NN	O	O

1	NN	O	O
1	NN	O	O
.	NN	O	O

ADCA	NN	O	B-Disease
type	NN	O	I-Disease
I	NN	O	I-Disease
patients	NN	O	O
from	NN	O	O
families	NN	O	O
segregating	NN	O	O
SCA3	NN	O	B-Disease
share	NN	O	O
clinical	NN	O	O
features	NN	O	O
in	NN	O	O
common	NN	O	O
with	NN	O	O
those	NN	O	O
with	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
of	NN	O	O
which	NN	O	O
maps	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
region	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
segregating	NN	O	O
in	NN	O	O
each	NN	O	O
of	NN	O	O
three	NN	O	O
French	NN	O	O
ADCA	NN	O	B-Disease
type	NN	O	I-Disease
I	NN	O	I-Disease
kindreds	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
French	NN	O	O
family	NN	O	O
with	NN	O	O
neuropathological	NN	O	O
findings	NN	O	O
suggesting	NN	O	O
the	NN	O	O
ataxochoreic	NN	O	O
form	NN	O	O
of	NN	O	O
dentatorubropallidoluysian	NN	O	B-Disease
atrophy	NN	O	I-Disease
carries	NN	O	O
an	NN	O	O
expanded	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
sequence	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
locus	NN	O	O
as	NN	O	O
that	NN	O	O
for	NN	O	O
MJD	NN	O	B-Disease
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
shows	NN	O	O
a	NN	O	O
strong	NN	O	O
negative	NN	O	O
correlation	NN	O	O
between	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
expanded	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
and	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
of	NN	O	O
clinical	NN	O	O
disease	NN	O	O
.	NN	O	O

Instability	NN	O	O
of	NN	O	O
the	NN	O	O
expanded	NN	O	O
triplet	NN	O	O
repeat	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
sex	NN	O	O
of	NN	O	O
the	NN	O	O
parent	NN	O	O
transmitting	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

Evidence	NN	O	O
was	NN	O	O
found	NN	O	O
for	NN	O	O
somatic	NN	O	O
and	NN	O	O
gonadal	NN	O	O
mosaicism	NN	O	O
for	NN	O	O
alleles	NN	O	O
carrying	NN	O	O
expanded	NN	O	O
trinucleotide	NN	O	O
repeats	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
DM20	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
in	NN	O	O
two	NN	O	O
brothers	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
rare	NN	O	B-Disease
,	NN	O	I-Disease
sex	NN	O	I-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
,	NN	O	I-Disease
dysmyelinating	NN	O	I-Disease
disease	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
central	NN	O	I-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
that	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
myelin	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Only	NN	O	O
25	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
studied	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
have	NN	O	O
exonic	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
underlying	NN	O	O
cause	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
patients	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

The	NN	O	O
PLP	NN	O	O
gene	NN	O	O
encodes	NN	O	O
two	NN	O	O
major	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
transcripts	NN	O	O
called	NN	O	O
PLP	NN	O	O
and	NN	O	O
DM20	NN	O	O
.	NN	O	O

PLP	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
is	NN	O	O
specifically	NN	O	O
expressed	NN	O	O
in	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
tissue	NN	O	O
,	NN	O	O
whereas	NN	O	O
DM20	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
cardiac	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
tissues	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
2	NN	O	O
brothers	NN	O	O
with	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
who	NN	O	O
exhibited	NN	O	O
no	NN	O	O
detectable	NN	O	O
exonic	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
PLP	NN	O	O
gene	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
RNA	NN	O	O
from	NN	O	O
these	NN	O	O
cells	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
DM20	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
is	NN	O	O
elevated	NN	O	O
sixfold	NN	O	O
relative	NN	O	O
to	NN	O	O
male	NN	O	O
control	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
.	NN	O	O

An	NN	O	O
unrelated	NN	O	O
female	NN	O	O
carrier	NN	O	O
,	NN	O	O
also	NN	O	O
with	NN	O	O
no	NN	O	O
detectable	NN	O	O
exonic	NN	O	O
mutation	NN	O	O
,	NN	O	O
showed	NN	O	O
a	NN	O	O
threefold	NN	O	O
increase	NN	O	O
in	NN	O	O
DM20	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
in	NN	O	O
cultured	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
some	NN	O	O
patients	NN	O	O
,	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
PLP	NN	O	O
gene	NN	O	O
transcripts	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
increase	NN	O	O
of	NN	O	O
DM20	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
in	NN	O	O
females	NN	O	O
,	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
affected	NN	O	O
males	NN	O	O
,	NN	O	O
can	NN	O	O
identify	NN	O	O
a	NN	O	O
female	NN	O	O
carrier	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
congenital	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
are	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
thrombocytopenia	NN	O	B-Disease
,	NN	O	O
small	NN	O	O
platelets	NN	O	O
,	NN	O	O
eczema	NN	O	B-Disease
,	NN	O	O
recurrent	NN	O	O
infections	NN	O	O
,	NN	O	O
and	NN	O	O
immunodeficiency	NN	O	B-Disease
.	NN	O	O

Besides	NN	O	O
the	NN	O	O
classic	NN	O	O
WAS	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
X	NN	O	I-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
(	NN	O	O
XLT	NN	O	B-Disease
)	NN	O	O
who	NN	O	O
have	NN	O	O
small	NN	O	O
platelets	NN	O	O
but	NN	O	O
only	NN	O	O
transient	NN	O	O
eczema	NN	O	B-Disease
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
and	NN	O	O
minimal	NN	O	O
immune	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Because	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
WAS	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
sequenced	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
possible	NN	O	O
to	NN	O	O
correlate	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
phenotypes	NN	O	O
with	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
mutations	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
fingerprinting	NN	O	O
screening	NN	O	O
technique	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
approximate	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
13	NN	O	O
unrelated	NN	O	O
WAS	NN	O	B-Disease
patients	NN	O	O
with	NN	O	O
mild	NN	O	O
to	NN	O	O
severe	NN	O	O
clinical	NN	O	O
symptoms	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
cDNA	NN	O	O
and	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
obtained	NN	O	O
from	NN	O	O
patient	NN	O	O
-	NN	O	O
derived	NN	O	O
cell	NN	O	O
lines	NN	O	O
showed	NN	O	O
12	NN	O	O
unique	NN	O	O
mutations	NN	O	O
distributed	NN	O	O
throughout	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
including	NN	O	O
insertions	NN	O	O
,	NN	O	O
deletions	NN	O	O
,	NN	O	O
and	NN	O	O
point	NN	O	O
mutations	NN	O	O
resulting	NN	O	O
in	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
,	NN	O	O
termination	NN	O	O
,	NN	O	O
exon	NN	O	O
skipping	NN	O	O
,	NN	O	O
or	NN	O	O
splicing	NN	O	O
defects	NN	O	O
.	NN	O	O

Of	NN	O	O
4	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
XLT	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
3	NN	O	O
had	NN	O	O
missense	NN	O	O
mutations	NN	O	O
affecting	NN	O	O
exon	NN	O	O
2	NN	O	O
and	NN	O	O
1	NN	O	O
had	NN	O	O
a	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
affecting	NN	O	O
exon	NN	O	O
9	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
classic	NN	O	O
WAS	NN	O	B-Disease
had	NN	O	O
more	NN	O	O
complex	NN	O	O
mutations	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
termination	NN	O	O
codons	NN	O	O
,	NN	O	O
frameshift	NN	O	O
,	NN	O	O
and	NN	O	O
early	NN	O	O
termination	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
XLT	NN	O	B-Disease
and	NN	O	O
WAS	NN	O	B-Disease
are	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
severe	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
complex	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Myotonia	NN	O	B-Disease
levior	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
chloride	NN	O	B-Disease
channel	NN	O	I-Disease
disorder	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
group	NN	O	O
of	NN	O	O
dominant	NN	O	B-Disease
non	NN	O	I-Disease
-	NN	O	I-Disease
dystrophic	NN	O	I-Disease
myotonias	NN	O	I-Disease
,	NN	O	O
comprising	NN	O	O
disorders	NN	O	O
characterized	NN	O	O
by	NN	O	O
clinically	NN	O	O
similar	NN	O	O
forms	NN	O	O
of	NN	O	O
myogenic	NN	O	O
muscle	NN	O	O
stiffness	NN	O	O
,	NN	O	O
is	NN	O	O
genetically	NN	O	O
inhomogeneous	NN	O	O
.	NN	O	O

Dominant	NN	O	B-Disease
myotonia	NN	O	I-Disease
congenita	NN	O	I-Disease
(	NN	O	O
Thomsens	NN	O	B-Disease
disease	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
CLCN1	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
major	NN	O	O
muscle	NN	O	O
chloride	NN	O	O
channel	NN	O	O
,	NN	O	O
localized	NN	O	O
on	NN	O	O
chromosome	NN	O	O
7q35	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
dominant	NN	O	B-Disease
myotonias	NN	O	I-Disease
sensitive	NN	O	O
to	NN	O	O
potassium	NN	O	O
are	NN	O	O
caused	NN	O	O
by	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
SCN4A	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
alpha	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
sodium	NN	O	O
channel	NN	O	O
.	NN	O	O

No	NN	O	O
linkage	NN	O	O
or	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
data	NN	O	O
are	NN	O	O
as	NN	O	O
yet	NN	O	O
available	NN	O	O
on	NN	O	O
myotonia	NN	O	B-Disease
levior	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
milder	NN	O	O
symptoms	NN	O	O
and	NN	O	O
later	NN	O	O
onset	NN	O	O
of	NN	O	O
myotonia	NN	O	B-Disease
than	NN	O	O
in	NN	O	O
Thomsens	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
muscle	NN	O	B-Disease
hypertrophy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
CLCN1	NN	O	O
Gln	NN	O	O
-	NN	O	O
552	NN	O	O
-	NN	O	O
Arg	NN	O	O
substitution	NN	O	O
for	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
dominant	NN	O	O
inheritance	NN	O	O
previously	NN	O	O
diagnosed	NN	O	O
to	NN	O	O
have	NN	O	O
myotonia	NN	O	B-Disease
levior	NN	O	I-Disease
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
disorder	NN	O	O
appears	NN	O	O
as	NN	O	O
a	NN	O	O
variant	NN	O	O
of	NN	O	O
Thomsens	NN	O	B-Disease
disease	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
mutations	NN	O	O
leading	NN	O	O
to	NN	O	O
low	NN	O	O
clinical	NN	O	O
expressivity	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
a	NN	O	O
novel	NN	O	O
Ile	NN	O	O
-	NN	O	O
290	NN	O	O
-	NN	O	O
Met	NN	O	O
CLCN1	NN	O	O
mutation	NN	O	O
for	NN	O	O
a	NN	O	O
typical	NN	O	O
Thomsen	NN	O	O
pedigree	NN	O	O
.	NN	O	O

In	NN	O	O
another	NN	O	O
family	NN	O	O
previously	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
Thomsens	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
unexpectedly	NN	O	O
found	NN	O	O
a	NN	O	O
CLCN1	NN	O	O
14	NN	O	O
bp	NN	O	O
deletion	NN	O	O
known	NN	O	O
to	NN	O	O
cause	NN	O	O
recessive	NN	O	B-Disease
myotonia	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
a	NN	O	O
rare	NN	O	O
Trp	NN	O	O
-	NN	O	O
118	NN	O	O
-	NN	O	O
Gly	NN	O	O
polymorphism	NN	O	O
.	NN	O	O
.	NN	O	O

Southern	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
a	NN	O	O
large	NN	O	O
deletion	NN	O	O
at	NN	O	O
the	NN	O	O
hypoxanthine	NN	O	O
phosphoribosyltransferase	NN	O	O
locus	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Whole	NN	O	O
genomic	NN	O	O
hprt	NN	O	O
clones	NN	O	O
were	NN	O	O
used	NN	O	O
in	NN	O	O
Southern	NN	O	O
analysis	NN	O	O
to	NN	O	O
screen	NN	O	O
the	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
hprt	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
that	NN	O	O
includes	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
HPRT	NN	O	B-Disease
enzyme	NN	O	I-Disease
deficiency	NN	O	I-Disease
causal	NN	O	O
to	NN	O	O
Lesch	NN	O	B-Disease
-	NN	O	I-Disease
Nyhan	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
5	NN	O	O
kb	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
deletion	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
its	NN	O	O
endpoints	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
and	NN	O	O
third	NN	O	O
introns	NN	O	O
.	NN	O	O

The	NN	O	O
probes	NN	O	O
identified	NN	O	O
the	NN	O	O
carrier	NN	O	O
status	NN	O	O
of	NN	O	O
female	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
aided	NN	O	O
by	NN	O	O
an	NN	O	O
RFLP	NN	O	O
carried	NN	O	O
by	NN	O	O
the	NN	O	O
mothers	NN	O	O
normal	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
.	NN	O	O
.	NN	O	O

Characterisation	NN	O	O
of	NN	O	O
molecular	NN	O	O
defects	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
amelogenesis	NN	O	I-Disease
imperfecta	NN	O	I-Disease
(	NN	O	O
AIH1	NN	O	O
)	NN	O	O
.	NN	O	O

Amelogenins	NN	O	O
are	NN	O	O
an	NN	O	O
heterogenous	NN	O	O
family	NN	O	O
of	NN	O	O
proteins	NN	O	O
produced	NN	O	O
by	NN	O	O
ameloblasts	NN	O	O
of	NN	O	O
the	NN	O	O
enamel	NN	O	O
organ	NN	O	O
during	NN	O	O
tooth	NN	O	O
development	NN	O	O
.	NN	O	O

Disturbances	NN	O	O
of	NN	O	O
enamel	NN	O	O
formation	NN	O	O
occur	NN	O	O
in	NN	O	O
amelogenesis	NN	O	B-Disease
imperfecta	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
clinically	NN	O	O
heterogenous	NN	O	O
group	NN	O	O
of	NN	O	O
inherited	NN	O	B-Disease
disorders	NN	O	I-Disease
characterised	NN	O	O
by	NN	O	O
defective	NN	O	O
enamel	NN	O	O
biomineralisation	NN	O	O
.	NN	O	O

An	NN	O	O
amelogenin	NN	O	O
gene	NN	O	O
,	NN	O	O
AMGX	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
short	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
(	NN	O	O
Xp22	NN	O	O
.	NN	O	O
1	NN	O	O
-	NN	O	O
p22	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathology	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
amelogenesis	NN	O	I-Disease
imperfecta	NN	O	I-Disease
(	NN	O	O
AIH1	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
three	NN	O	O
families	NN	O	O
exhibiting	NN	O	O
AIH1	NN	O	B-Disease
and	NN	O	O
screened	NN	O	O
the	NN	O	O
AMGX	NN	O	O
gene	NN	O	O
for	NN	O	O
mutations	NN	O	O
using	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Three	NN	O	O
novel	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
T	NN	O	O
substitution	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
T	NN	O	O
substitution	NN	O	O
and	NN	O	O
single	NN	O	O
cytosine	NN	O	O
deletion	NN	O	O
in	NN	O	O
exon	NN	O	O
6	NN	O	O
,	NN	O	O
confirming	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
extensive	NN	O	O
allelic	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
this	NN	O	O
condition	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
family	NN	O	O
-	NN	O	O
specific	NN	O	O
mutations	NN	O	O
will	NN	O	O
enable	NN	O	O
early	NN	O	O
identification	NN	O	O
of	NN	O	O
affected	NN	O	O
individuals	NN	O	O
and	NN	O	O
correlation	NN	O	O
of	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
with	NN	O	O
genotype	NN	O	O
will	NN	O	O
facilitate	NN	O	O
an	NN	O	O
objective	NN	O	O
system	NN	O	O
of	NN	O	O
disease	NN	O	O
classification	NN	O	O
.	NN	O	O

Frequency	NN	O	O
of	NN	O	O
exon	NN	O	O
15	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
442D	NN	O	O
:	NN	O	O
G	NN	O	O
)	NN	O	O
in	NN	O	O
cholesteryl	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
gene	NN	O	O
in	NN	O	O
hyperalphalipoproteinemic	NN	O	B-Disease
Japanese	NN	O	O
subjects	NN	O	O
.	NN	O	O

Cholesteryl	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
(	NN	O	O
CETP	NN	O	O
)	NN	O	O
transfers	NN	O	O
cholesteryl	NN	O	O
ester	NN	O	O
from	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
to	NN	O	O
apo	NN	O	O
B	NN	O	O
-	NN	O	O
containing	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

The	NN	O	O
hyperalphalipoproteinemia	NN	O	B-Disease
caused	NN	O	O
by	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
fairly	NN	O	O
common	NN	O	O
in	NN	O	O
Japan	NN	O	O
and	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
CETP	NN	O	O
gene	NN	O	O
is	NN	O	O
the	NN	O	O
splicing	NN	O	O
defect	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	O
14	NN	O	O
,	NN	O	O
the	NN	O	O
allelic	NN	O	O
frequency	NN	O	O
of	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
0	NN	O	O
.	NN	O	O

0049	NN	O	O
in	NN	O	O
the	NN	O	O
Japanese	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
15	NN	O	O
of	NN	O	O
the	NN	O	O
CETP	NN	O	O
gene	NN	O	O
(	NN	O	O
442D	NN	O	O
G	NN	O	O
)	NN	O	O
,	NN	O	O
showing	NN	O	O
a	NN	O	O
dominant	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
CETP	NN	O	O
activity	NN	O	O
and	NN	O	O
HDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
this	NN	O	O
new	NN	O	O
mutation	NN	O	O
in	NN	O	O
Japanese	NN	O	O
hyperalphalipoproteinemic	NN	O	B-Disease
(	NN	O	O
HDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
100	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
)	NN	O	O
subjects	NN	O	O
.	NN	O	O

A	NN	O	O
rapid	NN	O	O
and	NN	O	O
easy	NN	O	O
screening	NN	O	O
method	NN	O	O
for	NN	O	O
this	NN	O	O
new	NN	O	O
mutation	NN	O	O
was	NN	O	O
developed	NN	O	O
using	NN	O	O
a	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
-	NN	O	O
mediated	NN	O	O
site	NN	O	O
-	NN	O	O
directed	NN	O	O
mutagenesis	NN	O	O
.	NN	O	O

Among	NN	O	O
117	NN	O	O
Japanese	NN	O	O
hyperalphalipoproteinemic	NN	O	B-Disease
subjects	NN	O	O
(	NN	O	O
HDL	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
;	NN	O	O
116	NN	O	O
.	NN	O	O
7	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
16	NN	O	O
.	NN	O	O
5	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
,	NN	O	O
mean	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
S	NN	O	O
.	NN	O	O
D	NN	O	O
.	NN	O	O
)	NN	O	O
without	NN	O	O
the	NN	O	O
intron	NN	O	O
14	NN	O	O
splice	NN	O	O
defect	NN	O	O
,	NN	O	O
three	NN	O	O
homozygotes	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
5	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
34	NN	O	O
heterozygotes	NN	O	O
(	NN	O	O
29	NN	O	O
.	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
442D	NN	O	O
G	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
relative	NN	O	O
allelic	NN	O	O
frequency	NN	O	O
of	NN	O	O
this	NN	O	O
mutation	NN	O	O
was	NN	O	O
calculated	NN	O	O
to	NN	O	O
be	NN	O	O
0	NN	O	O
.	NN	O	O

17	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
homozygotes	NN	O	O
for	NN	O	O
the	NN	O	O
442D	NN	O	O
G	NN	O	O
mutation	NN	O	O
was	NN	O	O
the	NN	O	O
patient	NN	O	O
previously	NN	O	O
described	NN	O	O
by	NN	O	O
us	NN	O	O
as	NN	O	O
having	NN	O	O
hyperalphalipoproteinemia	NN	O	B-Disease
with	NN	O	O
corneal	NN	O	B-Disease
opacity	NN	O	I-Disease
and	NN	O	O
coronary	NN	O	B-Disease
heart	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
was	NN	O	O
the	NN	O	O
first	NN	O	O
reported	NN	O	O
subject	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
who	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
atherosclerotic	NN	O	B-Disease
symptoms	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
homozygous	NN	O	O
subjects	NN	O	O
,	NN	O	O
CETP	NN	O	O
activity	NN	O	O
ranged	NN	O	O
from	NN	O	O
37	NN	O	O
%	NN	O	O
to	NN	O	O
62	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
value	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
results	NN	O	O
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
transient	NN	O	O
expression	NN	O	O
experiment	NN	O	O
previously	NN	O	O
reported	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
specific	NN	O	O
activity	NN	O	O
of	NN	O	O
CETP	NN	O	O
was	NN	O	O
not	NN	O	O
as	NN	O	O
low	NN	O	O
as	NN	O	O
expected	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

Mucopolysaccharidosis	NN	O	B-Disease
type	NN	O	I-Disease
IVA	NN	O	I-Disease
:	NN	O	O
common	NN	O	O
double	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
acetylgalactosamine	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
sulfatase	NN	O	O
gene	NN	O	O
(	NN	O	O
GALNS	NN	O	O
)	NN	O	O
.	NN	O	O

Mucopolysaccharidosis	NN	O	B-Disease
IVA	NN	O	I-Disease
(	NN	O	O
MPS	NN	O	B-Disease
IVA	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
in	NN	O	I-Disease
N	NN	O	I-Disease
-	NN	O	I-Disease
acetylgalactosamine	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
sulfatase	NN	O	I-Disease
(	NN	O	O
GALNS	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
two	NN	O	O
separate	NN	O	O
deletions	NN	O	O
of	NN	O	O
nearly	NN	O	O
8	NN	O	O
.	NN	O	O

0	NN	O	O
and	NN	O	O
6	NN	O	O
.	NN	O	O

0	NN	O	O
kb	NN	O	O
in	NN	O	O
the	NN	O	O
GALNS	NN	O	O
gene	NN	O	O
,	NN	O	O
including	NN	O	O
some	NN	O	O
exons	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
Alu	NN	O	O
repetitive	NN	O	O
elements	NN	O	O
near	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
the	NN	O	O
8	NN	O	O
.	NN	O	O

0	NN	O	O
-	NN	O	O
kb	NN	O	O
deletion	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
deletion	NN	O	O
resulted	NN	O	O
from	NN	O	O
an	NN	O	O
Alu	NN	O	O
-	NN	O	O
Alu	NN	O	O
recombination	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
6	NN	O	O
.	NN	O	O

0	NN	O	O
-	NN	O	O
kb	NN	O	O
deletion	NN	O	O
involved	NN	O	O
illegitimate	NN	O	O
recombinational	NN	O	O
events	NN	O	O
between	NN	O	O
incomplete	NN	O	O
short	NN	O	O
direct	NN	O	O
repeats	NN	O	O
of	NN	O	O
8	NN	O	O
bp	NN	O	O
at	NN	O	O
deletion	NN	O	O
breakpoints	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
rearrangement	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
heteroallelic	NN	O	O
state	NN	O	O
in	NN	O	O
four	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
documentation	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
double	NN	O	O
deletion	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
cluster	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
eight	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
10	NN	O	O
breast	NN	O	B-Disease
/	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
,	NN	O	O
including	NN	O	O
1	NN	O	O
family	NN	O	O
with	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Genetic	NN	O	O
epidemiological	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
approximately	NN	O	O
one	NN	O	O
half	NN	O	O
of	NN	O	O
early	NN	O	O
onset	NN	O	O
familial	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
familial	NN	O	B-Disease
breast	NN	O	I-Disease
/	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
recent	NN	O	O
cloning	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
allows	NN	O	O
for	NN	O	O
the	NN	O	O
direct	NN	O	O
detection	NN	O	O
of	NN	O	O
mutations	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
feasibility	NN	O	O
of	NN	O	O
presymptomatic	NN	O	O
screening	NN	O	O
for	NN	O	O
cancer	NN	O	B-Disease
susceptibility	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
one	NN	O	O
affected	NN	O	O
individual	NN	O	O
from	NN	O	O
each	NN	O	O
of	NN	O	O
24	NN	O	O
families	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
cases	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
or	NN	O	I-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
using	NN	O	O
SSCP	NN	O	O
assays	NN	O	O
.	NN	O	O

Variant	NN	O	O
SSCP	NN	O	O
bands	NN	O	O
were	NN	O	O
subcloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Allele	NN	O	O
-	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
hybridization	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
verify	NN	O	O
sequence	NN	O	O
changes	NN	O	O
and	NN	O	O
to	NN	O	O
screen	NN	O	O
DNA	NN	O	O
from	NN	O	O
control	NN	O	O
individuals	NN	O	O
.	NN	O	O

Six	NN	O	O
frameshift	NN	O	O
and	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
10	NN	O	O
different	NN	O	O
families	NN	O	O
.	NN	O	O

A	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
male	NN	O	O
proband	NN	O	O
affected	NN	O	O
with	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
40	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
who	NN	O	O
developed	NN	O	O
intra	NN	O	B-Disease
-	NN	O	I-Disease
abdominal	NN	O	I-Disease
carcinomatosis	NN	O	I-Disease
1	NN	O	O
year	NN	O	O
after	NN	O	O
prophylactic	NN	O	O
oophorectomy	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
throughout	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
only	NN	O	O
one	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
more	NN	O	O
than	NN	O	O
a	NN	O	O
single	NN	O	O
family	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
that	NN	O	O
inherited	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
can	NN	O	O
occur	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
a	NN	O	O
wide	NN	O	O
array	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggests	NN	O	O
that	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
screening	NN	O	O
test	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
will	NN	O	O
be	NN	O	O
technically	NN	O	O
challenging	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
not	NN	O	O
previously	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
also	NN	O	O
illustrates	NN	O	O
the	NN	O	O
potential	NN	O	O
difficulties	NN	O	O
of	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
for	NN	O	O
individuals	NN	O	O
known	NN	O	O
to	NN	O	O
carry	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Natural	NN	O	O
selection	NN	O	O
of	NN	O	O
hemi	NN	O	O
-	NN	O	O
and	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
Africa	NN	O	O
by	NN	O	O
resistance	NN	O	O
to	NN	O	O
severe	NN	O	O
malaria	NN	O	B-Disease
.	NN	O	O

Glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
enzymopathy	NN	O	B-Disease
of	NN	O	O
humans	NN	O	O
,	NN	O	O
affects	NN	O	O
over	NN	O	O
400	NN	O	O
million	NN	O	O
people	NN	O	O
.	NN	O	O

The	NN	O	O
geographical	NN	O	O
correlation	NN	O	O
of	NN	O	O
its	NN	O	O
distribution	NN	O	O
with	NN	O	O
the	NN	O	O
historical	NN	O	O
endemicity	NN	O	O
of	NN	O	O
malaria	NN	O	B-Disease
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
disorder	NN	O	O
has	NN	O	O
risen	NN	O	O
in	NN	O	O
frequency	NN	O	O
through	NN	O	O
natural	NN	O	O
selection	NN	O	O
by	NN	O	O
malaria	NN	O	B-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
attempts	NN	O	O
to	NN	O	O
confirm	NN	O	O
that	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
protective	NN	O	O
in	NN	O	O
case	NN	O	O
-	NN	O	O
control	NN	O	O
studies	NN	O	O
of	NN	O	O
malaria	NN	O	B-Disease
have	NN	O	O
yielded	NN	O	O
conflicting	NN	O	O
results	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
for	NN	O	O
this	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
is	NN	O	O
unclear	NN	O	O
whether	NN	O	O
both	NN	O	O
male	NN	O	O
hemizygotes	NN	O	O
and	NN	O	O
female	NN	O	O
heterozygotes	NN	O	O
are	NN	O	O
protected	NN	O	O
or	NN	O	O
,	NN	O	O
as	NN	O	O
frequently	NN	O	O
suggested	NN	O	O
,	NN	O	O
only	NN	O	O
females	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
how	NN	O	O
much	NN	O	O
protection	NN	O	O
may	NN	O	O
be	NN	O	O
afforded	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
two	NN	O	O
large	NN	O	O
case	NN	O	O
-	NN	O	O
control	NN	O	O
studies	NN	O	O
of	NN	O	O
over	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
African	NN	O	O
children	NN	O	O
,	NN	O	O
the	NN	O	O
common	NN	O	O
African	NN	O	O
form	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
G6PD	NN	O	B-Disease
A	NN	O	I-Disease
-	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
46	NN	O	O
-	NN	O	O
58	NN	O	O
%	NN	O	O
reduction	NN	O	O
in	NN	O	O
risk	NN	O	O
of	NN	O	O
severe	NN	O	O
malaria	NN	O	B-Disease
for	NN	O	O
both	NN	O	O
female	NN	O	O
heterozygotes	NN	O	O
and	NN	O	O
male	NN	O	O
hemizygotes	NN	O	O
.	NN	O	O

A	NN	O	O
mathematical	NN	O	O
model	NN	O	O
incorporating	NN	O	O
the	NN	O	O
measured	NN	O	O
selective	NN	O	O
advantage	NN	O	O
against	NN	O	O
malaria	NN	O	B-Disease
suggests	NN	O	O
that	NN	O	O
a	NN	O	O
counterbalancing	NN	O	O
selective	NN	O	O
disadvantage	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
this	NN	O	O
enzyme	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
has	NN	O	O
retarded	NN	O	O
its	NN	O	O
rise	NN	O	O
in	NN	O	O
frequency	NN	O	O
in	NN	O	O
malaria	NN	O	B-Disease
-	NN	O	O
endemic	NN	O	O
regions	NN	O	O
.	NN	O	O

Although	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
now	NN	O	O
regarded	NN	O	O
as	NN	O	O
a	NN	O	O
generally	NN	O	O
benign	NN	O	O
disorder	NN	O	O
,	NN	O	O
in	NN	O	O
earlier	NN	O	O
environmental	NN	O	O
conditions	NN	O	O
it	NN	O	O
could	NN	O	O
have	NN	O	O
been	NN	O	O
significantly	NN	O	O
disadvantageous	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
alveolar	NN	O	B-Disease
rhabdomyosarcoma	NN	O	I-Disease
PAX3	NN	O	O
/	NN	O	O
FKHR	NN	O	O
fusion	NN	O	O
protein	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
.	NN	O	O

Chimeric	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
created	NN	O	O
by	NN	O	O
gene	NN	O	O
fusions	NN	O	O
as	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
several	NN	O	O
pathologically	NN	O	O
disparate	NN	O	O
solid	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
PAX3	NN	O	O
/	NN	O	O
FKHR	NN	O	O
fusion	NN	O	O
gene	NN	O	O
,	NN	O	O
formed	NN	O	O
by	NN	O	O
a	NN	O	O
t	NN	O	O
(	NN	O	O
2	NN	O	O
;	NN	O	O
13	NN	O	O
)	NN	O	O
(	NN	O	O
q35	NN	O	O
;	NN	O	O
q14	NN	O	O
)	NN	O	O
in	NN	O	O
alveolar	NN	O	B-Disease
rhabdomyosarcoma	NN	O	I-Disease
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
hybrid	NN	O	O
protein	NN	O	O
that	NN	O	O
contains	NN	O	O
both	NN	O	O
PAX3	NN	O	O
DNA	NN	O	O
binding	NN	O	O
domains	NN	O	O
,	NN	O	O
the	NN	O	O
paired	NN	O	O
box	NN	O	O
and	NN	O	O
homeodomain	NN	O	O
,	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
bisected	NN	O	O
DNA	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
FKHR	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
forkhead	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
PAX3	NN	O	O
and	NN	O	O
PAX3	NN	O	O
/	NN	O	O
FKHR	NN	O	O
display	NN	O	O
similar	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
identical	NN	O	O
transactivation	NN	O	O
activities	NN	O	O
when	NN	O	O
tested	NN	O	O
with	NN	O	O
model	NN	O	O
Pax	NN	O	O
recognition	NN	O	O
sequences	NN	O	O
.	NN	O	O

No	NN	O	O
functional	NN	O	O
role	NN	O	O
could	NN	O	O
be	NN	O	O
ascribed	NN	O	O
solely	NN	O	O
to	NN	O	O
the	NN	O	O
residual	NN	O	O
FKHR	NN	O	O
binding	NN	O	O
domain	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
fusion	NN	O	O
protein	NN	O	O
,	NN	O	O
but	NN	O	O
FKHR	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
contribute	NN	O	O
a	NN	O	O
strong	NN	O	O
carboxyl	NN	O	O
terminal	NN	O	O
activation	NN	O	O
domain	NN	O	O
replacing	NN	O	O
the	NN	O	O
one	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
unrearranged	NN	O	O
PAX3	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
native	NN	O	O
PAX3	NN	O	O
/	NN	O	O
FKHR	NN	O	O
protein	NN	O	O
present	NN	O	O
in	NN	O	O
tumor	NN	O	B-Disease
cells	NN	O	O
with	NN	O	O
this	NN	O	O
translocation	NN	O	O
has	NN	O	O
transcriptional	NN	O	O
characteristics	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
expressed	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
PAX3	NN	O	O
/	NN	O	O
FKHR	NN	O	O
hybrid	NN	O	O
protein	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
in	NN	O	O
a	NN	O	O
sequence	NN	O	O
specific	NN	O	O
manner	NN	O	O
and	NN	O	O
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
artificial	NN	O	O
reporter	NN	O	O
genes	NN	O	O
suggests	NN	O	O
that	NN	O	O
its	NN	O	O
aberrant	NN	O	O
expression	NN	O	O
could	NN	O	O
subvert	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
programs	NN	O	O
that	NN	O	O
normally	NN	O	O
control	NN	O	O
the	NN	O	O
growth	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
survival	NN	O	O
of	NN	O	O
primitive	NN	O	O
myogenic	NN	O	O
precursors	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
p16INK4a	NN	O	O
-	NN	O	O
insensitive	NN	O	O
CDK4	NN	O	O
mutant	NN	O	O
targeted	NN	O	O
by	NN	O	O
cytolytic	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	O
melanoma	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
mutated	NN	O	O
cyclin	NN	O	O
-	NN	O	O
dependent	NN	O	O
kinase	NN	O	O
4	NN	O	O
(	NN	O	O
CDK4	NN	O	O
)	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
-	NN	O	O
specific	NN	O	O
antigen	NN	O	O
recognized	NN	O	O
by	NN	O	O
HLA	NN	O	O
-	NN	O	O
A2	NN	O	O
.	NN	O	O

1	NN	O	O
-	NN	O	O
restricted	NN	O	O
autologous	NN	O	O
cytolytic	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
(	NN	O	O
CTLs	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	O
melanoma	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
mutated	NN	O	O
CDK4	NN	O	O
allele	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
autologous	NN	O	O
cultured	NN	O	O
melanoma	NN	O	B-Disease
cells	NN	O	O
and	NN	O	O
metastasis	NN	O	O
tissue	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
,	NN	O	O
an	NN	O	O
arginine	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
cysteine	NN	O	O
exchange	NN	O	O
at	NN	O	O
residue	NN	O	O
24	NN	O	O
,	NN	O	O
was	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
CDK4	NN	O	O
peptide	NN	O	O
recognized	NN	O	O
by	NN	O	O
CTLs	NN	O	O
and	NN	O	O
prevented	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
CDK4	NN	O	O
inhibitor	NN	O	O
p16INK4a	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
p21	NN	O	O
or	NN	O	O
of	NN	O	O
p27KIP1	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
additional	NN	O	O
melanoma	NN	O	B-Disease
among	NN	O	O
28	NN	O	O
melanomas	NN	O	B-Disease
analyzed	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
CDK4	NN	O	O
can	NN	O	O
create	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
-	NN	O	O
specific	NN	O	O
antigen	NN	O	O
and	NN	O	O
can	NN	O	O
disrupt	NN	O	O
the	NN	O	O
cell	NN	O	O
-	NN	O	O
cycle	NN	O	O
regulation	NN	O	O
exerted	NN	O	O
by	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
p16INK4a	NN	O	O
.	NN	O	O
.	NN	O	O

Rapid	NN	O	O
detection	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
by	NN	O	O
the	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
.	NN	O	O

More	NN	O	O
than	NN	O	O
75	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
reported	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
gene	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
result	NN	O	O
in	NN	O	O
truncated	NN	O	O
proteins	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
(	NN	O	O
PTT	NN	O	O
)	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
exon	NN	O	O
11	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
61	NN	O	O
%	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

In	NN	O	O
45	NN	O	O
patients	NN	O	O
from	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
we	NN	O	O
found	NN	O	O
six	NN	O	O
novel	NN	O	O
mutations	NN	O	O
two	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
insertions	NN	O	O
,	NN	O	O
three	NN	O	O
small	NN	O	O
deletions	NN	O	O
(	NN	O	O
1	NN	O	O
-	NN	O	O
5	NN	O	O
bp	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
identified	NN	O	O
two	NN	O	O
unrelated	NN	O	O
families	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
amplify	NN	O	O
the	NN	O	O
remaining	NN	O	O
coding	NN	O	O
region	NN	O	O
by	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
using	NN	O	O
lymphocyte	NN	O	O
RNA	NN	O	O
.	NN	O	O

Combined	NN	O	O
with	NN	O	O
PTT	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
aberrantly	NN	O	O
spliced	NN	O	O
products	NN	O	O
affecting	NN	O	O
exons	NN	O	O
5	NN	O	O
and	NN	O	O
6	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
linked	NN	O	O
families	NN	O	O
examined	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
promises	NN	O	O
to	NN	O	O
become	NN	O	O
a	NN	O	O
valuable	NN	O	O
technique	NN	O	O
in	NN	O	O
detecting	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Cloning	NN	O	O
of	NN	O	O
human	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
coenzyme	NN	O	O
A	NN	O	O
dehydrogenase	NN	O	O
and	NN	O	O
molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
its	NN	O	O
deficiency	NN	O	O
in	NN	O	O
two	NN	O	O
patients	NN	O	O
.	NN	O	O

Two	NN	O	O
overlapping	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
991	NN	O	O
bp	NN	O	O
and	NN	O	O
736	NN	O	O
bp	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
encoding	NN	O	O
the	NN	O	O
precursor	NN	O	O
of	NN	O	O
human	NN	O	O
mitochondrial	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
coenzyme	NN	O	O
A	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
VLCAD	NN	O	O
)	NN	O	O
were	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

The	NN	O	O
cDNA	NN	O	O
inserts	NN	O	O
of	NN	O	O
these	NN	O	O
clones	NN	O	O
together	NN	O	O
encompass	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
177	NN	O	O
bases	NN	O	O
,	NN	O	O
encoding	NN	O	O
the	NN	O	O
entire	NN	O	O
protein	NN	O	O
of	NN	O	O
655	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
40	NN	O	O
-	NN	O	O
amino	NN	O	O
acid	NN	O	O
leader	NN	O	O
peptide	NN	O	O
and	NN	O	O
a	NN	O	O
615	NN	O	O
-	NN	O	O
amino	NN	O	O
acid	NN	O	O
mature	NN	O	O
polypeptide	NN	O	O
.	NN	O	O

PCR	NN	O	O
-	NN	O	O
amplified	NN	O	O
VLCAD	NN	O	O
cDNAs	NN	O	O
were	NN	O	O
sequenced	NN	O	O
in	NN	O	O
cultured	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
two	NN	O	O
VLCAD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
patients	NN	O	O
,	NN	O	O
a	NN	O	O
105	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
encompassing	NN	O	O
bases	NN	O	O
1078	NN	O	O
-	NN	O	O
1182	NN	O	O
in	NN	O	O
VLCAD	NN	O	O
cDNA	NN	O	O
was	NN	O	O
identified	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
seems	NN	O	O
to	NN	O	O
occur	NN	O	O
due	NN	O	O
to	NN	O	O
exon	NN	O	O
skipping	NN	O	O
during	NN	O	O
processing	NN	O	O
of	NN	O	O
VLCAD	NN	O	O
pre	NN	O	O
-	NN	O	O
mRNA	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
causing	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Quantitative	NN	O	O
cDNA	NN	O	O
expression	NN	O	O
of	NN	O	O
normal	NN	O	O
human	NN	O	O
VLCAD	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
using	NN	O	O
vaccinia	NN	O	O
viral	NN	O	O
system	NN	O	O
,	NN	O	O
which	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
normal	NN	O	I-Disease
VLCAD	NN	O	I-Disease
protein	NN	O	I-Disease
causes	NN	O	O
impaired	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
fibroblasts	NN	O	O
.	NN	O	O

In	NN	O	O
patient	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
raising	NN	O	O
VLCAD	NN	O	O
activity	NN	O	O
to	NN	O	O
approximately	NN	O	O
20	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
control	NN	O	O
fibroblast	NN	O	O
activity	NN	O	O
raised	NN	O	O
palmitic	NN	O	O
acid	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
flux	NN	O	O
to	NN	O	O
the	NN	O	O
level	NN	O	O
found	NN	O	O
in	NN	O	O
control	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
offer	NN	O	O
important	NN	O	O
information	NN	O	O
for	NN	O	O
the	NN	O	O
rational	NN	O	O
design	NN	O	O
of	NN	O	O
future	NN	O	O
somatic	NN	O	O
gene	NN	O	O
therapy	NN	O	O
for	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Brain	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Abnormal	NN	O	O
amplification	NN	O	O
of	NN	O	O
a	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
on	NN	O	O
chromosome	NN	O	O
19	NN	O	O
is	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Expansion	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
has	NN	O	O
been	NN	O	O
correlated	NN	O	O
with	NN	O	O
severity	NN	O	O
of	NN	O	O
several	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
extensive	NN	O	O
cognitive	NN	O	O
testing	NN	O	O
,	NN	O	O
cerebral	NN	O	O
magnetic	NN	O	O
resonance	NN	O	O
imaging	NN	O	O
(	NN	O	O
MRI	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
in	NN	O	O
28	NN	O	O
cases	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
to	NN	O	O
determine	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
molecular	NN	O	O
defect	NN	O	O
and	NN	O	O
brain	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Performance	NN	O	O
in	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
cognitive	NN	O	O
tests	NN	O	O
was	NN	O	O
pathological	NN	O	O
in	NN	O	O
10	NN	O	O
cases	NN	O	O
.	NN	O	O

Fourteen	NN	O	O
patients	NN	O	O
had	NN	O	O
subcortical	NN	O	O
white	NN	O	B-Disease
matter	NN	O	I-Disease
lesions	NN	O	I-Disease
on	NN	O	O
MRI	NN	O	O
,	NN	O	O
14	NN	O	O
had	NN	O	O
cerebral	NN	O	B-Disease
atrophy	NN	O	I-Disease
.	NN	O	O

Amplification	NN	O	O
of	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
showed	NN	O	O
a	NN	O	O
strong	NN	O	O
correlation	NN	O	O
with	NN	O	O
cognitive	NN	O	O
test	NN	O	O
deficits	NN	O	O
when	NN	O	O
exceeding	NN	O	O
a	NN	O	O
length	NN	O	O
of	NN	O	O
over	NN	O	O
1000	NN	O	O
trinucleotides	NN	O	O
.	NN	O	O

MRI	NN	O	O
lesions	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
impaired	NN	O	O
psychometric	NN	O	O
performance	NN	O	O
,	NN	O	O
but	NN	O	O
MRI	NN	O	O
and	NN	O	O
molecular	NN	O	O
findings	NN	O	O
were	NN	O	O
only	NN	O	O
weakly	NN	O	O
related	NN	O	O
.	NN	O	O

Disease	NN	O	O
duration	NN	O	O
influenced	NN	O	O
the	NN	O	O
appearance	NN	O	O
and	NN	O	O
amount	NN	O	O
of	NN	O	O
white	NN	O	B-Disease
matter	NN	O	I-Disease
lesions	NN	O	I-Disease
on	NN	O	O
MRI	NN	O	O
.	NN	O	O

Quantification	NN	O	O
of	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
size	NN	O	O
may	NN	O	O
allow	NN	O	O
an	NN	O	O
early	NN	O	O
estimate	NN	O	O
on	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
brain	NN	O	O
involvement	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
;	NN	O	O
cognitive	NN	O	B-Disease
dysfunction	NN	O	I-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
white	NN	O	B-Disease
matter	NN	O	I-Disease
lesions	NN	O	I-Disease
and	NN	O	O
cerebral	NN	O	B-Disease
atrophy	NN	O	I-Disease
later	NN	O	O
on	NN	O	O
in	NN	O	O
the	NN	O	O
course	NN	O	O
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
in	NN	O	O
patients	NN	O	O
with	NN	O	O
different	NN	O	O
clinical	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
peroxisomal	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
encoding	NN	O	O
a	NN	O	O
peroxisomal	NN	O	O
membrane	NN	O	O
transporter	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
entire	NN	O	O
protein	NN	O	O
-	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
by	NN	O	O
reverse	NN	O	O
-	NN	O	O
transcription	NN	O	O
PCR	NN	O	O
,	NN	O	O
SSCP	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
in	NN	O	O
five	NN	O	O
patients	NN	O	O
with	NN	O	O
different	NN	O	O
clinical	NN	O	O
expression	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
and	NN	O	O
in	NN	O	O
their	NN	O	O
female	NN	O	O
relatives	NN	O	O
;	NN	O	O
these	NN	O	O
clinical	NN	O	O
expressions	NN	O	O
were	NN	O	O
cerebral	NN	O	O
childhood	NN	O	O
ALD	NN	O	B-Disease
,	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
"	NN	O	O
Addison	NN	O	B-Disease
disease	NN	O	I-Disease
only	NN	O	I-Disease
"	NN	O	O
(	NN	O	O
ADO	NN	O	B-Disease
)	NN	O	O
phenotype	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
three	NN	O	O
patients	NN	O	O
exhibiting	NN	O	O
the	NN	O	O
classical	NN	O	O
picture	NN	O	O
of	NN	O	O
severe	NN	O	O
childhood	NN	O	O
ALD	NN	O	B-Disease
we	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
gene	NN	O	O
a	NN	O	O
38	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
that	NN	O	O
causes	NN	O	O
a	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
,	NN	O	O
a	NN	O	O
3	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
an	NN	O	O
amino	NN	O	O
acid	NN	O	O
in	NN	O	O
the	NN	O	O
ATP	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
protein	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
of	NN	O	O
AMN	NN	O	B-Disease
,	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
codon	NN	O	O
212	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
a	NN	O	O
second	NN	O	O
site	NN	O	O
mutation	NN	O	O
at	NN	O	O
codon	NN	O	O
178	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
ADO	NN	O	B-Disease
phenotype	NN	O	O
revealed	NN	O	O
a	NN	O	O
further	NN	O	O
missense	NN	O	O
mutation	NN	O	O
at	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
position	NN	O	O
in	NN	O	O
the	NN	O	O
ALDP	NN	O	O
/	NN	O	O
PMP70	NN	O	O
comparison	NN	O	O
.	NN	O	O

The	NN	O	O
disruptive	NN	O	O
nature	NN	O	O
of	NN	O	O
two	NN	O	O
mutations	NN	O	O
(	NN	O	O
i	NN	O	O
.	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
the	NN	O	O
frameshift	NN	O	O
and	NN	O	O
the	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
)	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
biochemically	NN	O	O
proved	NN	O	O
childhood	NN	O	O
ALD	NN	O	B-Disease
and	NN	O	O
AMN	NN	O	B-Disease
further	NN	O	O
strongly	NN	O	O
supports	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
alterations	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
.	NN	O	O

Since	NN	O	O
the	NN	O	O
current	NN	O	O
biochemical	NN	O	O
techniques	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
carrier	NN	O	O
detection	NN	O	O
in	NN	O	O
affected	NN	O	O
families	NN	O	O
lack	NN	O	O
sufficient	NN	O	O
reliability	NN	O	O
,	NN	O	O
our	NN	O	O
procedure	NN	O	O
described	NN	O	O
for	NN	O	O
systematic	NN	O	O
mutation	NN	O	O
scanning	NN	O	O
is	NN	O	O
also	NN	O	O
capable	NN	O	O
of	NN	O	O
improving	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
and	NN	O	O
prenatal	NN	O	O
diagnosis	NN	O	O

Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
same	NN	O	O
patient	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
first	NN	O	O
patient	NN	O	O
identified	NN	O	O
with	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
family	NN	O	O
of	NN	O	O
the	NN	O	O
propositus	NN	O	O
had	NN	O	O
a	NN	O	O
strong	NN	O	O
history	NN	O	O
of	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
intrafamilial	NN	O	O
pathological	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
responsible	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
between	NN	O	O
the	NN	O	O
mildly	NN	O	O
affected	NN	O	O
mother	NN	O	O
(	NN	O	O
160	NN	O	O
repeats	NN	O	O
;	NN	O	O
normal	NN	O	O
27	NN	O	O
repeats	NN	O	O
)	NN	O	O
and	NN	O	O
her	NN	O	O
more	NN	O	O
severely	NN	O	O
affected	NN	O	O
son	NN	O	O
(	NN	O	O
650	NN	O	O
repeats	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
his	NN	O	O
sister	NN	O	O
(	NN	O	O
650	NN	O	O
repeats	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
propositus	NN	O	O
was	NN	O	O
an	NN	O	O
isolated	NN	O	O
case	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
with	NN	O	O
marked	NN	O	O
dystrophin	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
muscle	NN	O	O
biopsy	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
still	NN	O	O
ambulatory	NN	O	O
post	NN	O	O
age	NN	O	O
16	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
could	NN	O	O
interfere	NN	O	O
to	NN	O	O
some	NN	O	O
extent	NN	O	O
with	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

However	NN	O	O
,	NN	O	O
other	NN	O	O
interpretations	NN	O	O
are	NN	O	O
possible	NN	O	O
.	NN	O	O

Twelve	NN	O	O
percent	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
revertant	NN	O	O
fibers	NN	O	O
as	NN	O	O
observed	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
could	NN	O	O
be	NN	O	O
sufficient	NN	O	O
to	NN	O	O
ameliorate	NN	O	O
typical	NN	O	O
DMD	NN	O	B-Disease
clinical	NN	O	O
severity	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
patient	NN	O	O
may	NN	O	O
present	NN	O	O
a	NN	O	O
somatic	NN	O	O
mosaic	NN	O	O
.	NN	O	O

The	NN	O	O
pathophysiological	NN	O	O
interactions	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
unlinked	NN	O	O
disorders	NN	O	O
are	NN	O	O
discussed	NN	O	O
at	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
histopathological	NN	O	O
levels	NN	O	O
.	NN	O	O
.	NN	O	O

Age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
as	NN	O	O
an	NN	O	O
indicator	NN	O	O
of	NN	O	O
eligibility	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
DNA	NN	O	O
testing	NN	O	O
in	NN	O	O
familial	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
searched	NN	O	O
for	NN	O	O
criteria	NN	O	O
that	NN	O	O
could	NN	O	O
indicate	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
prior	NN	O	O
probability	NN	O	O
of	NN	O	O
being	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
/	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
locus	NN	O	O
BRCA1	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17	NN	O	O
.	NN	O	O

To	NN	O	O
this	NN	O	O
end	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
a	NN	O	O
linkage	NN	O	O
study	NN	O	O
with	NN	O	O
59	NN	O	O
consecutively	NN	O	O
collected	NN	O	O
Dutch	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
,	NN	O	O
including	NN	O	O
16	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
case	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
used	NN	O	O
an	NN	O	O
intake	NN	O	O
cut	NN	O	O
-	NN	O	O
off	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
first	NN	O	O
-	NN	O	O
degree	NN	O	O
relatives	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
at	NN	O	O
any	NN	O	O
age	NN	O	O
.	NN	O	O

Significant	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
was	NN	O	O
found	NN	O	O
only	NN	O	O
among	NN	O	O
the	NN	O	O
13	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
families	NN	O	O
with	NN	O	O
a	NN	O	O
mean	NN	O	O
age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
less	NN	O	O
than	NN	O	O
45	NN	O	O
years	NN	O	O
.	NN	O	O

An	NN	O	O
unexpectedly	NN	O	O
low	NN	O	O
proportion	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
were	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
which	NN	O	O
could	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
Dutch	NN	O	O
population	NN	O	O
.	NN	O	O

Given	NN	O	O
the	NN	O	O
expected	NN	O	O
logistical	NN	O	O
problems	NN	O	O
in	NN	O	O
clinical	NN	O	O
management	NN	O	O
now	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
an	NN	O	O
interim	NN	O	O
period	NN	O	O
in	NN	O	O
which	NN	O	O
only	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
strong	NN	O	O
positive	NN	O	O
family	NN	O	O
history	NN	O	O
for	NN	O	O
early	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
will	NN	O	O
be	NN	O	O
offered	NN	O	O
BRCA1	NN	O	O
mutation	NN	O	O
testing	NN	O	O
.	NN	O	O
.	NN	O	O

Linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
26	NN	O	O
Canadian	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
breast	NN	O	I-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
26	NN	O	O
Canadian	NN	O	O
families	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
markers	NN	O	O
flanking	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
15	NN	O	O
families	NN	O	O
that	NN	O	O
contain	NN	O	O
cases	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
94	NN	O	O
%	NN	O	O
were	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
overall	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
in	NN	O	O
the	NN	O	O
group	NN	O	O
of	NN	O	O
10	NN	O	O
families	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
without	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
genetic	NN	O	O
recombinant	NN	O	O
in	NN	O	O
a	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
family	NN	O	O
indicates	NN	O	O
a	NN	O	O
placement	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
telomeric	NN	O	O
to	NN	O	O
D17S776	NN	O	O
,	NN	O	O
and	NN	O	O
helps	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
assignment	NN	O	O
of	NN	O	O
the	NN	O	O
cancer	NN	O	B-Disease
susceptibility	NN	O	O
gene	NN	O	O
.	NN	O	O

Other	NN	O	O
cancers	NN	O	B-Disease
of	NN	O	O
interest	NN	O	O
that	NN	O	O
appeared	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
-	NN	O	O
linked	NN	O	O
families	NN	O	O
included	NN	O	O
primary	NN	O	B-Disease
peritoneal	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
cancer	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fallopian	NN	O	I-Disease
tube	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
malignant	NN	O	B-Disease
melanoma	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Structural	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
region	NN	O	O
of	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
genes	NN	O	O
reveals	NN	O	O
conservation	NN	O	O
of	NN	O	O
putative	NN	O	O
promoter	NN	O	O
region	NN	O	O
and	NN	O	O
di	NN	O	O
-	NN	O	O
and	NN	O	O
trinucleotide	NN	O	O
polymorphisms	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
cloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
the	NN	O	O
murine	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
and	NN	O	O
shown	NN	O	O
that	NN	O	O
it	NN	O	O
maps	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
5	NN	O	O
within	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
conserved	NN	O	O
synteny	NN	O	O
with	NN	O	O
human	NN	O	O
chromosome	NN	O	O
4p16	NN	O	O
.	NN	O	O

3	NN	O	O
3	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
detailed	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
promoter	NN	O	O
and	NN	O	O
the	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
genomic	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
(	NN	O	O
Hdh	NN	O	O
)	NN	O	O
and	NN	O	O
human	NN	O	O
HD	NN	O	B-Disease
genes	NN	O	O
encompassing	NN	O	O
the	NN	O	O
first	NN	O	O
five	NN	O	O
exons	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
these	NN	O	O
two	NN	O	O
genes	NN	O	O
share	NN	O	O
identical	NN	O	O
exon	NN	O	O
boundaries	NN	O	O
,	NN	O	O
but	NN	O	O
have	NN	O	O
different	NN	O	O
-	NN	O	O
size	NN	O	O
introns	NN	O	O
.	NN	O	O

Two	NN	O	O
dinucleotide	NN	O	O
(	NN	O	O
CT	NN	O	O
)	NN	O	O
and	NN	O	O
one	NN	O	O
trinucleotide	NN	O	O
intronic	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
Hdh	NN	O	O
and	NN	O	O
an	NN	O	O
intronic	NN	O	O
CA	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
940	NN	O	O
-	NN	O	O
bp	NN	O	O
sequence	NN	O	O
5	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	O
translation	NN	O	O
start	NN	O	O
site	NN	O	O
reveals	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
region	NN	O	O
(	NN	O	O
78	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
nucleotide	NN	O	O
identity	NN	O	O
)	NN	O	O
between	NN	O	O
Hdh	NN	O	O
and	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
from	NN	O	O
nucleotide	NN	O	O
-	NN	O	O
56	NN	O	O
to	NN	O	O
-	NN	O	O
206	NN	O	O
(	NN	O	O
of	NN	O	O
Hdh	NN	O	O
)	NN	O	O
.	NN	O	O

Neither	NN	O	O
Hdh	NN	O	O
nor	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
have	NN	O	O
typical	NN	O	O
TATA	NN	O	O
or	NN	O	O
CCAAT	NN	O	O
elements	NN	O	O
,	NN	O	O
but	NN	O	O
both	NN	O	O
show	NN	O	O
one	NN	O	O
putative	NN	O	O
AP2	NN	O	O
binding	NN	O	O
site	NN	O	O
and	NN	O	O
numerous	NN	O	O
potential	NN	O	O
Sp1	NN	O	O
binding	NN	O	O
sites	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
sequence	NN	O	O
identity	NN	O	O
between	NN	O	O
Hdh	NN	O	O
and	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
for	NN	O	O
approximately	NN	O	O
200	NN	O	O
bp	NN	O	O
5	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	O
translation	NN	O	O
start	NN	O	O
site	NN	O	O
indicates	NN	O	O
that	NN	O	O
these	NN	O	O
sequences	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O

The	NN	O	O
protein	NN	O	O
deficient	NN	O	O
in	NN	O	O
Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
phosphatidylinositol	NN	O	O
-	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
bisphosphate	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
.	NN	O	O

Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
oculocerebrorenal	NN	O	B-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
-	NN	O	O
encoded	NN	O	O
OCRL	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
OCRL	NN	O	O
protein	NN	O	O
is	NN	O	O
51	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
inositol	NN	O	O
polyphosphate	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
II	NN	O	O
(	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
II	NN	O	O
)	NN	O	O
from	NN	O	O
human	NN	O	O
platelets	NN	O	O
over	NN	O	O
a	NN	O	O
span	NN	O	O
of	NN	O	O
744	NN	O	O
aa	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
OCRL	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
similar	NN	O	O
enzyme	NN	O	O
.	NN	O	O

We	NN	O	O
engineered	NN	O	O
a	NN	O	O
construct	NN	O	O
of	NN	O	O
the	NN	O	O
OCRL	NN	O	O
cDNA	NN	O	O
that	NN	O	O
encodes	NN	O	O
amino	NN	O	O
acids	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
platelet	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
for	NN	O	O
expression	NN	O	O
in	NN	O	O
baculovirus	NN	O	O
-	NN	O	O
infected	NN	O	O
Sf9	NN	O	O
insect	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
cDNA	NN	O	O
encodes	NN	O	O
aa	NN	O	O
264	NN	O	O
-	NN	O	O
968	NN	O	O
of	NN	O	O
the	NN	O	O
OCRL	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
recombinant	NN	O	O
protein	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
catalyze	NN	O	O
the	NN	O	O
reactions	NN	O	O
also	NN	O	O
carried	NN	O	O
out	NN	O	O
by	NN	O	O
platelet	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
II	NN	O	O
.	NN	O	O

Thus	NN	O	O
OCRL	NN	O	O
converts	NN	O	O
inositol	NN	O	O
1	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
trisphosphate	NN	O	O
to	NN	O	O
inositol	NN	O	O
1	NN	O	O
,	NN	O	O
4	NN	O	O
-	NN	O	O
bisphosphate	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
converts	NN	O	O
inositol	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
tetrakisphosphate	NN	O	O
to	NN	O	O
inositol	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4	NN	O	O
-	NN	O	O
trisphosphate	NN	O	O
.	NN	O	O

Most	NN	O	O
important	NN	O	O
,	NN	O	O
the	NN	O	O
enzyme	NN	O	O
converts	NN	O	O
phosphatidylinositol	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
bisphosphate	NN	O	O
to	NN	O	O
phosphatidylinositol	NN	O	O
4	NN	O	O
-	NN	O	O
phosphate	NN	O	O
.	NN	O	O

The	NN	O	O
relative	NN	O	O
ability	NN	O	O
of	NN	O	O
OCRL	NN	O	O
to	NN	O	O
catalyze	NN	O	O
the	NN	O	O
three	NN	O	O
reactions	NN	O	O
is	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
II	NN	O	O
and	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
another	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
isoenzyme	NN	O	O
from	NN	O	O
platelets	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
I	NN	O	O
.	NN	O	O

The	NN	O	O
recombinant	NN	O	O
OCRL	NN	O	O
protein	NN	O	O
hydrolyzes	NN	O	O
the	NN	O	O
phospholipid	NN	O	O
substrate	NN	O	O
10	NN	O	O
-	NN	O	O
to	NN	O	O
30	NN	O	O
-	NN	O	O
fold	NN	O	O
better	NN	O	O
than	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
II	NN	O	O
,	NN	O	O
and	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
I	NN	O	O
does	NN	O	O
not	NN	O	O
cleave	NN	O	O
the	NN	O	O
lipid	NN	O	O
at	NN	O	O
all	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
OCRL	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
phosphatidylinositol	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
bisphosphate	NN	O	O
5	NN	O	O
-	NN	O	O
phosphatase	NN	O	O
in	NN	O	O
OCRL	NN	O	O
-	NN	O	O
expressing	NN	O	O
Sf9	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
OCRL	NN	O	O
is	NN	O	O
mainly	NN	O	O
a	NN	O	O
lipid	NN	O	O
phosphatase	NN	O	O
that	NN	O	O
may	NN	O	O
control	NN	O	O
cellular	NN	O	O
levels	NN	O	O
of	NN	O	O
a	NN	O	O
critical	NN	O	O
metabolite	NN	O	O
,	NN	O	O
phosphatidylinositol	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
-	NN	O	O
bisphosphate	NN	O	O
.	NN	O	O

Deficiency	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
apparently	NN	O	O
causes	NN	O	O
the	NN	O	O
protean	NN	O	O
manifestations	NN	O	O
of	NN	O	O
Lowe	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

New	NN	O	O
founder	NN	O	O
haplotypes	NN	O	O
at	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
locus	NN	O	O
in	NN	O	O
southern	NN	O	O
Africa	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
between	NN	O	O
normal	NN	O	O
alleles	NN	O	O
at	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
and	NN	O	O
two	NN	O	O
nearby	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
the	NN	O	O
myotonin	NN	O	O
protein	NN	O	O
kinase	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
Alu	NN	O	O
insertion	NN	O	O
/	NN	O	O
deletion	NN	O	O
polymorphism	NN	O	O
and	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
kinase	NN	O	O
(	NN	O	O
DMK	NN	O	O
)	NN	O	O
(	NN	O	O
G	NN	O	O
/	NN	O	O
T	NN	O	O
)	NN	O	O
intron	NN	O	O
9	NN	O	O
/	NN	O	O
HinfI	NN	O	O
polymorphism	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
analyzed	NN	O	O
in	NN	O	O
South	NN	O	O
African	NN	O	O
Negroids	NN	O	O
,	NN	O	O
a	NN	O	O
population	NN	O	O
in	NN	O	O
which	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

South	NN	O	O
African	NN	O	O
Negroids	NN	O	O
have	NN	O	O
a	NN	O	O
CTG	NN	O	O
allelic	NN	O	O
distribution	NN	O	O
that	NN	O	O
is	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
in	NN	O	O
Caucasoids	NN	O	O
and	NN	O	O
Japanese	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
lengths	NN	O	O
of	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
19	NN	O	O
are	NN	O	O
very	NN	O	O
rare	NN	O	O
.	NN	O	O

The	NN	O	O
striking	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
between	NN	O	O
specific	NN	O	O
alleles	NN	O	O
at	NN	O	O
the	NN	O	O
Alu	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
Alu	NN	O	O
(	NN	O	O
ins	NN	O	O
)	NN	O	O
and	NN	O	O
Alu	NN	O	O
(	NN	O	O
del	NN	O	O
)	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
HinfI	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
HinfI	NN	O	O
-	NN	O	O
1	NN	O	O
and	NN	O	O
HinfI	NN	O	O
-	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
polymorphism	NN	O	O
seen	NN	O	O
in	NN	O	O
Caucasoid	NN	O	O
(	NN	O	O
Europeans	NN	O	O
and	NN	O	O
Canadians	NN	O	O
)	NN	O	O
populations	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
South	NN	O	O
African	NN	O	O
Negroid	NN	O	O
population	NN	O	O
.	NN	O	O

Numerous	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
not	NN	O	O
previously	NN	O	O
described	NN	O	O
in	NN	O	O
Europeans	NN	O	O
,	NN	O	O
were	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
found	NN	O	O
.	NN	O	O

It	NN	O	O
thus	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
only	NN	O	O
a	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
these	NN	O	O
"	NN	O	O
African	NN	O	O
"	NN	O	O
chromosomes	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
progenitors	NN	O	O
of	NN	O	O
all	NN	O	O
non	NN	O	O
-	NN	O	O
African	NN	O	O
peoples	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
support	NN	O	O
for	NN	O	O
the	NN	O	O
"	NN	O	O
out	NN	O	O
of	NN	O	O
Africa	NN	O	O
"	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
modern	NN	O	O
humans	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
rare	NN	O	O
ancestral	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
event	NN	O	O
may	NN	O	O
have	NN	O	O
occurred	NN	O	O
after	NN	O	O
the	NN	O	O
migration	NN	O	O
from	NN	O	O
Africa	NN	O	O
,	NN	O	O
hence	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
in	NN	O	O
sub	NN	O	O
-	NN	O	O
Saharan	NN	O	O
Negroid	NN	O	O
peoples	NN	O	O
.	NN	O	O
.	NN	O	O

Discordant	NN	O	O
clinical	NN	O	O
outcome	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
relatives	NN	O	O
showing	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
>	NN	O	O
700	NN	O	O
repeats	NN	O	O
.	NN	O	O

A	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
family	NN	O	O
is	NN	O	O
described	NN	O	O
in	NN	O	O
which	NN	O	O
discordant	NN	O	O
DM	NN	O	B-Disease
phenotypes	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
children	NN	O	O
of	NN	O	O
two	NN	O	O
affected	NN	O	O
sisters	NN	O	O
with	NN	O	O
similar	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
and	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
congenital	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
early	NN	O	O
severe	NN	O	O
childhood	NN	O	O
and	NN	O	O
later	NN	O	O
childhood	NN	O	O
onset	NN	O	O
DM	NN	O	B-Disease
coexist	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
strengthens	NN	O	O
the	NN	O	O
limited	NN	O	O
ability	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
number	NN	O	O
analysis	NN	O	O
in	NN	O	O
predicting	NN	O	O
genotype	NN	O	O
-	NN	O	O
phenotype	NN	O	O
correlations	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O
.	NN	O	O

Purification	NN	O	O
of	NN	O	O
human	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
coenzyme	NN	O	O
A	NN	O	O
dehydrogenase	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
its	NN	O	O
deficiency	NN	O	O
in	NN	O	O
seven	NN	O	O
patients	NN	O	O
.	NN	O	O

Mitochondrial	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
coenzyme	NN	O	O
A	NN	O	O
dehydrogenase	NN	O	O
(	NN	O	O
VLCAD	NN	O	O
)	NN	O	O
was	NN	O	O
purified	NN	O	O
from	NN	O	O
human	NN	O	O
liver	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
masses	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	O
enzyme	NN	O	O
and	NN	O	O
the	NN	O	O
subunit	NN	O	O
were	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
154	NN	O	O
and	NN	O	O
70	NN	O	O
kD	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
enzyme	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
catalyze	NN	O	O
the	NN	O	O
major	NN	O	O
part	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
palmitoylcoenzyme	NN	O	O
A	NN	O	O
dehydrogenation	NN	O	O
in	NN	O	O
liver	NN	O	O
,	NN	O	O
heart	NN	O	O
,	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
,	NN	O	O
and	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
(	NN	O	O
89	NN	O	O
-	NN	O	O
97	NN	O	O
,	NN	O	O
86	NN	O	O
-	NN	O	O
99	NN	O	O
,	NN	O	O
96	NN	O	O
-	NN	O	O
99	NN	O	O
,	NN	O	O
and	NN	O	O
78	NN	O	O
-	NN	O	O
87	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

Skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
26	NN	O	O
patients	NN	O	O
suspected	NN	O	O
of	NN	O	O
having	NN	O	O
a	NN	O	O
disorder	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
VLCAD	NN	O	O
protein	NN	O	O
using	NN	O	O
immunoblotting	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
of	NN	O	O
them	NN	O	O
contained	NN	O	O
undetectable	NN	O	O
or	NN	O	O
trace	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
.	NN	O	O

The	NN	O	O
seven	NN	O	O
deficient	NN	O	O
fibroblast	NN	O	O
lines	NN	O	O
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
measuring	NN	O	O
acyl	NN	O	O
-	NN	O	O
coenzyme	NN	O	O
A	NN	O	O
dehydrogenation	NN	O	O
activities	NN	O	O
,	NN	O	O
overall	NN	O	O
palmitic	NN	O	O
acid	NN	O	O
oxidation	NN	O	O
,	NN	O	O
and	NN	O	O
VLCAD	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
using	NN	O	O
pulse	NN	O	O
-	NN	O	O
chase	NN	O	O
,	NN	O	O
further	NN	O	O
confirming	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggested	NN	O	O
the	NN	O	O
heterogenous	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
causing	NN	O	O
the	NN	O	O
deficiency	NN	O	O
in	NN	O	O
the	NN	O	O
seven	NN	O	O
patients	NN	O	O
.	NN	O	O

Clinically	NN	O	O
,	NN	O	O
all	NN	O	O
patients	NN	O	O
with	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
exhibited	NN	O	O
cardiac	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

At	NN	O	O
least	NN	O	O
four	NN	O	O
of	NN	O	O
them	NN	O	O
presented	NN	O	O
with	NN	O	O
hypertrophic	NN	O	B-Disease
cardiomyopathy	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
frequency	NN	O	O
(	NN	O	O
>	NN	O	O
57	NN	O	O
%	NN	O	O
)	NN	O	O
was	NN	O	O
much	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
other	NN	O	O
disorders	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acid	NN	O	O
oxidation	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
accompanied	NN	O	O
by	NN	O	O
cardiac	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
infants	NN	O	O
.	NN	O	O
.	NN	O	O

Hereditary	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fifth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
in	NN	O	O
man	NN	O	O
.	NN	O	O

II	NN	O	O
.	NN	O	O

Biological	NN	O	O
properties	NN	O	O
of	NN	O	O
C5	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
human	NN	O	O
serum	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
known	NN	O	O
human	NN	O	O
kindred	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
deficiency	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
fifth	NN	O	I-Disease
component	NN	O	I-Disease
of	NN	O	I-Disease
complement	NN	O	I-Disease
(	NN	O	O
C5	NN	O	O
)	NN	O	O
was	NN	O	O
documented	NN	O	O
in	NN	O	O
the	NN	O	O
accompanying	NN	O	O
report	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
examines	NN	O	O
several	NN	O	O
biological	NN	O	O
properties	NN	O	O
of	NN	O	O
C5	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
(	NN	O	O
C5D	NN	O	B-Disease
)	NN	O	O
human	NN	O	O
serum	NN	O	O
,	NN	O	O
particularly	NN	O	O
sera	NN	O	O
obtained	NN	O	O
from	NN	O	O
two	NN	O	O
C5D	NN	O	B-Disease
homozygotes	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
,	NN	O	O
who	NN	O	O
has	NN	O	O
inactive	NN	O	O
systemic	NN	O	B-Disease
lupus	NN	O	I-Disease
erythematosus	NN	O	I-Disease
is	NN	O	O
completely	NN	O	O
lacking	NN	O	O
C5	NN	O	O
,	NN	O	O
while	NN	O	O
her	NN	O	O
healthy	NN	O	O
half	NN	O	O
-	NN	O	O
sister	NN	O	O
has	NN	O	O
1	NN	O	O
-	NN	O	O
2	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
levels	NN	O	O
.	NN	O	O

Both	NN	O	O
sera	NN	O	O
were	NN	O	O
severely	NN	O	O
impaired	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
generate	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
for	NN	O	O
normal	NN	O	O
human	NN	O	O
neutrophils	NN	O	O
upon	NN	O	O
incubation	NN	O	O
with	NN	O	O
aggregated	NN	O	O
human	NN	O	O
gamma	NN	O	O
-	NN	O	O
globulin	NN	O	O
or	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
endotoxin	NN	O	O
.	NN	O	O

This	NN	O	O
function	NN	O	O
was	NN	O	O
fully	NN	O	O
restored	NN	O	O
in	NN	O	O
the	NN	O	O
siblings	NN	O	O
serum	NN	O	O
,	NN	O	O
and	NN	O	O
substantially	NN	O	O
improved	NN	O	O
in	NN	O	O
the	NN	O	O
probands	NN	O	O
serum	NN	O	O
,	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
highly	NN	O	O
purified	NN	O	O
human	NN	O	O
C5	NN	O	O
to	NN	O	O
normal	NN	O	O
serum	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Sera	NN	O	O
from	NN	O	O
eight	NN	O	O
family	NN	O	O
members	NN	O	O
who	NN	O	O
were	NN	O	O
apparently	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
C5	NN	O	B-Disease
deficiency	NN	O	I-Disease
gave	NN	O	O
normal	NN	O	O
chemotactic	NN	O	O
scores	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
C5D	NN	O	B-Disease
serum	NN	O	O
to	NN	O	O
opsonize	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
(	NN	O	O
bakers	NN	O	O
yeast	NN	O	O
)	NN	O	O
or	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
for	NN	O	O
ingestion	NN	O	O
by	NN	O	O
normal	NN	O	O
neutrophils	NN	O	O
was	NN	O	O
completely	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
C5D	NN	O	B-Disease
serum	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
promoting	NN	O	O
normal	NN	O	O
phagocytosis	NN	O	O
and	NN	O	O
intracellular	NN	O	O
killing	NN	O	O
of	NN	O	O
Staphylococcus	NN	O	O
aureus	NN	O	O
.	NN	O	O

The	NN	O	O
probands	NN	O	O
serum	NN	O	O
was	NN	O	O
incapable	NN	O	O
of	NN	O	O
mediating	NN	O	O
lysis	NN	O	O
of	NN	O	O
erythrocytes	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
paroxysmal	NN	O	B-Disease
nocturnal	NN	O	I-Disease
hemoglobinuria	NN	O	I-Disease
in	NN	O	O
both	NN	O	O
the	NN	O	O
sucrose	NN	O	O
hemolysia	NN	O	O
and	NN	O	O
acid	NN	O	O
hemolysis	NN	O	O
tests	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
lacked	NN	O	O
bactericidal	NN	O	O
activity	NN	O	O
against	NN	O	O
sensitized	NN	O	O
or	NN	O	O
unsensitized	NN	O	O
Salmonella	NN	O	O
typhi	NN	O	O
.	NN	O	O

The	NN	O	O
siblings	NN	O	O
serum	NN	O	O
,	NN	O	O
containing	NN	O	O
only	NN	O	O
1	NN	O	O
-	NN	O	O
2	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
C5	NN	O	O
,	NN	O	O
effectively	NN	O	O
lysed	NN	O	O
S	NN	O	O
.	NN	O	O
typhi	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
at	NN	O	O
eightfold	NN	O	O
lower	NN	O	O
serum	NN	O	O
dilutions	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
normals	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
underscore	NN	O	O
the	NN	O	O
critical	NN	O	O
role	NN	O	O
of	NN	O	O
C5	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
and	NN	O	O
in	NN	O	O
cytolytic	NN	O	O
reactions	NN	O	O
,	NN	O	O
as	NN	O	O
opposed	NN	O	O
to	NN	O	O
a	NN	O	O
nonobligatory	NN	O	O
or	NN	O	O
minimal	NN	O	O
role	NN	O	O
in	NN	O	O
opsonization	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
for	NN	O	O
the	NN	O	O
organisms	NN	O	O
under	NN	O	O
study	NN	O	O
.	NN	O	O
.	NN	O	O

Human	NN	O	O
peroxisomal	NN	O	O
targeting	NN	O	O
signal	NN	O	O
-	NN	O	O
1	NN	O	O
receptor	NN	O	O
restores	NN	O	O
peroxisomal	NN	O	O
protein	NN	O	O
import	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
fatal	NN	O	O
peroxisomal	NN	O	B-Disease
disorders	NN	O	I-Disease
.	NN	O	O

Two	NN	O	O
peroxisomal	NN	O	O
targeting	NN	O	O
signals	NN	O	O
,	NN	O	O
PTS1	NN	O	O
and	NN	O	O
PTS2	NN	O	O
,	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
import	NN	O	O
of	NN	O	O
proteins	NN	O	O
into	NN	O	O
the	NN	O	O
peroxisome	NN	O	O
matrix	NN	O	O
.	NN	O	O

Human	NN	O	O
patients	NN	O	O
with	NN	O	O
fatal	NN	O	O
generalized	NN	O	O
peroxisomal	NN	O	B-Disease
deficiency	NN	O	I-Disease
disorders	NN	O	I-Disease
fall	NN	O	O
into	NN	O	O
at	NN	O	O
least	NN	O	O
nine	NN	O	O
genetic	NN	O	O
complementation	NN	O	O
groups	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
are	NN	O	O
deficient	NN	O	O
in	NN	O	O
the	NN	O	O
import	NN	O	O
of	NN	O	O
PTS1	NN	O	O
-	NN	O	O
containing	NN	O	O
proteins	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
causes	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
-	NN	O	O
import	NN	O	O
defect	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
human	NN	O	O
cDNA	NN	O	O
homologue	NN	O	O
(	NN	O	O
PTS1R	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
Pichia	NN	O	O
pastoris	NN	O	O
PAS8	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
PTS1	NN	O	O
receptor	NN	O	O
(	NN	O	O
McCollum	NN	O	O
,	NN	O	O
D	NN	O	O
.	NN	O	O
,	NN	O	O
E	NN	O	O
.	NN	O	O
Monosov	NN	O	O
,	NN	O	O
and	NN	O	O
S	NN	O	O
.	NN	O	O
Subramani	NN	O	O
.	NN	O	O
1993	NN	O	O
.	NN	O	O
J	NN	O	O
.	NN	O	O
Cell	NN	O	O
Biol	NN	O	O
.	NN	O	O
121	NN	O	O
761	NN	O	O
-	NN	O	O
774	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
PTS1R	NN	O	O
mRNA	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
human	NN	O	O
tissues	NN	O	O
examined	NN	O	O
.	NN	O	O

Antibodies	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
PTS1R	NN	O	O
recognize	NN	O	O
this	NN	O	O
protein	NN	O	O
in	NN	O	O
human	NN	O	O
,	NN	O	O
monkey	NN	O	O
,	NN	O	O
rat	NN	O	O
,	NN	O	O
and	NN	O	O
hamster	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
is	NN	O	O
localized	NN	O	O
mainly	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
but	NN	O	O
is	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
peroxisomes	NN	O	O
.	NN	O	O

Part	NN	O	O
of	NN	O	O
the	NN	O	O
peroxisomal	NN	O	O
PTS1R	NN	O	O
protein	NN	O	O
is	NN	O	O
tightly	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
peroxisomal	NN	O	O
membrane	NN	O	O
.	NN	O	O

Antibodies	NN	O	O
to	NN	O	O
PTS1R	NN	O	O
inhibit	NN	O	O
peroxisomal	NN	O	O
protein	NN	O	O
-	NN	O	O
import	NN	O	O
of	NN	O	O
PTS1	NN	O	O
-	NN	O	O
containing	NN	O	O
proteins	NN	O	O
in	NN	O	O
a	NN	O	O
permeabilized	NN	O	O
CHO	NN	O	O
cell	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
-	NN	O	O
translated	NN	O	O
PTS1R	NN	O	O
protein	NN	O	O
specifically	NN	O	O
binds	NN	O	O
a	NN	O	O
serine	NN	O	O
-	NN	O	O
lysine	NN	O	O
-	NN	O	O
leucine	NN	O	O
-	NN	O	O
peptide	NN	O	O
.	NN	O	O

A	NN	O	O
PAS8	NN	O	O
-	NN	O	O
PTS1R	NN	O	O
fusion	NN	O	O
protein	NN	O	O
complements	NN	O	O
the	NN	O	O
P	NN	O	O
.	NN	O	O
pastoris	NN	O	O
pas8	NN	O	O
mutant	NN	O	O
.	NN	O	O

The	NN	O	O
PTS1R	NN	O	O
cDNA	NN	O	O
also	NN	O	O
complements	NN	O	O
the	NN	O	O
PTS1	NN	O	B-Disease
protein	NN	O	I-Disease
-	NN	O	I-Disease
import	NN	O	I-Disease
defect	NN	O	I-Disease
in	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
patients	NN	O	O
-	NN	O	O
-	NN	O	O
belonging	NN	O	O
to	NN	O	O
complementation	NN	O	O
group	NN	O	O
two	NN	O	O
-	NN	O	O
-	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
neonatal	NN	O	B-Disease
adrenoleukodystrophy	NN	O	I-Disease
or	NN	O	O
Zellweger	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
PTS1R	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
chromosomal	NN	O	O
location	NN	O	O
where	NN	O	O
no	NN	O	O
other	NN	O	O
peroxisomal	NN	O	B-Disease
disorder	NN	O	I-Disease
genes	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
map	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
represent	NN	O	O
the	NN	O	O
only	NN	O	O
case	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-Disease
-	NN	O	I-Disease
import	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
human	NN	O	O
peroxisomal	NN	O	B-Disease
disorders	NN	O	I-Disease
is	NN	O	O
understood	NN	O	O
.	NN	O	O

Spectrum	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
:	NN	O	O
a	NN	O	O
study	NN	O	O
of	NN	O	O
232	NN	O	O
patients	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
and	NN	O	I-Disease
non	NN	O	I-Disease
hereditary	NN	O	I-Disease
retinoblastoma	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
confer	NN	O	O
hereditary	NN	O	O
predisposition	NN	O	O
to	NN	O	O
retinoblastoma	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
performed	NN	O	O
a	NN	O	O
mutation	NN	O	O
survey	NN	O	O
of	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
in	NN	O	O
232	NN	O	O
patients	NN	O	O
with	NN	O	O
hereditary	NN	O	B-Disease
or	NN	O	I-Disease
non	NN	O	I-Disease
hereditary	NN	O	I-Disease
retinoblastoma	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
systematically	NN	O	O
explored	NN	O	O
all	NN	O	O
27	NN	O	O
exons	NN	O	O
and	NN	O	O
flanking	NN	O	O
sequences	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
promotor	NN	O	O
.	NN	O	O

All	NN	O	O
types	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
are	NN	O	O
represented	NN	O	O
and	NN	O	O
are	NN	O	O
found	NN	O	O
unequally	NN	O	O
distributed	NN	O	O
along	NN	O	O
the	NN	O	O
RB1	NN	O	O
gene	NN	O	O
sequence	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
population	NN	O	O
we	NN	O	O
studied	NN	O	O
,	NN	O	O
exons	NN	O	O
3	NN	O	O
,	NN	O	O
8	NN	O	O
,	NN	O	O
18	NN	O	O
and	NN	O	O
19	NN	O	O
are	NN	O	O
preferentially	NN	O	O
altered	NN	O	O
.	NN	O	O

The	NN	O	O
range	NN	O	O
of	NN	O	O
frequency	NN	O	O
of	NN	O	O
detection	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
is	NN	O	O
about	NN	O	O
20	NN	O	O
%	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
other	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
inactivation	NN	O	O
of	NN	O	O
RB1	NN	O	O
should	NN	O	O
be	NN	O	O
involved	NN	O	O
.	NN	O	O

The	NN	O	O
spectrum	NN	O	O
of	NN	O	O
mutations	NN	O	O
presented	NN	O	O
here	NN	O	O
should	NN	O	O
help	NN	O	O
to	NN	O	O
improve	NN	O	O
the	NN	O	O
clinical	NN	O	O
management	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-Disease
and	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
leading	NN	O	O
to	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O
.	NN	O	O

Aniridia	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
cytogenetic	NN	O	O
rearrangements	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
position	NN	O	O
effect	NN	O	O
may	NN	O	O
cause	NN	O	O
the	NN	O	O
mutant	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Current	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
aniridia	NN	O	B-Disease
(	NN	O	O
absence	NN	O	B-Disease
of	NN	O	I-Disease
iris	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
loss	NN	O	O
of	NN	O	O
function	NN	O	O
of	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
maps	NN	O	O
to	NN	O	O
11p13	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
the	NN	O	O
further	NN	O	O
characterisation	NN	O	O
of	NN	O	O
two	NN	O	O
aniridia	NN	O	B-Disease
pedigrees	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
disease	NN	O	O
segregates	NN	O	O
with	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
which	NN	O	O
involve	NN	O	O
11p13	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
disrupt	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
three	NN	O	O
human	NN	O	O
YAC	NN	O	O
clones	NN	O	O
which	NN	O	O
encompass	NN	O	O
the	NN	O	O
PAX6	NN	O	O
locus	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
these	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
both	NN	O	O
cases	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
breakpoint	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
85	NN	O	O
kb	NN	O	O
distal	NN	O	O
of	NN	O	O
the	NN	O	O
3	NN	O	O
end	NN	O	O
of	NN	O	O
PAX6	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
of	NN	O	O
PAX6	NN	O	O
is	NN	O	O
apparently	NN	O	O
free	NN	O	O
of	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
on	NN	O	O
the	NN	O	O
rearranged	NN	O	O
chromosome	NN	O	O
11	NN	O	O
is	NN	O	O
in	NN	O	O
an	NN	O	O
inappropriate	NN	O	O
chromatin	NN	O	O
environment	NN	O	O
for	NN	O	O
normal	NN	O	O
expression	NN	O	O
and	NN	O	O
therefore	NN	O	O
that	NN	O	O
a	NN	O	O
position	NN	O	O
effect	NN	O	O
is	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanism	NN	O	O
of	NN	O	O
disease	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
.	NN	O	O
.	NN	O	O

Somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
in	NN	O	O
sporadic	NN	O	B-Disease
ovarian	NN	O	I-Disease
tumours	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q21	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
syndrome	NN	O	I-Disease
of	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
but	NN	O	O
no	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
tumours	NN	O	B-Disease
have	NN	O	O
yet	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
sporadic	NN	O	O
carcinogenesis	NN	O	O
,	NN	O	O
we	NN	O	O
analysed	NN	O	O
the	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
of	NN	O	O
tumour	NN	O	B-Disease
and	NN	O	O
normal	NN	O	O
fractions	NN	O	O
of	NN	O	O
47	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
using	NN	O	O
the	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
technique	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
describe	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
four	NN	O	O
tumours	NN	O	B-Disease
which	NN	O	O
also	NN	O	O
had	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
(	NN	O	O
LOH	NN	O	O
)	NN	O	O
at	NN	O	O
a	NN	O	O
BRCA1	NN	O	O
intragenic	NN	O	O
marker	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
a	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
mechanism	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
;	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
and	NN	O	O
LOH	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
inactivation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Decreased	NN	O	O
expression	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
accelerates	NN	O	O
growth	NN	O	O
and	NN	O	O
is	NN	O	O
often	NN	O	O
present	NN	O	O
during	NN	O	O
sporadic	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
progression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
characterized	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
familial	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
gene	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
in	NN	O	O
cases	NN	O	O
of	NN	O	O
non	NN	O	B-Disease
-	NN	O	I-Disease
hereditary	NN	O	I-Disease
(	NN	O	I-Disease
sporadic	NN	O	I-Disease
)	NN	O	I-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
antisense	NN	O	O
inhibition	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
on	NN	O	O
the	NN	O	O
proliferative	NN	O	O
rate	NN	O	O
of	NN	O	O
mammary	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
are	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
during	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
carcinoma	NN	O	B-Disease
in	NN	O	I-Disease
situ	NN	O	I-Disease
to	NN	O	O
invasive	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Experimental	NN	O	O
inhibition	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
expression	NN	O	O
with	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
produced	NN	O	O
accelerated	NN	O	O
growth	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
malignant	NN	O	O
mammary	NN	O	O
cells	NN	O	O
,	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
non	NN	O	O
-	NN	O	O
mammary	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
may	NN	O	O
normally	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
mammary	NN	O	O
epithelial	NN	O	O
cell	NN	O	O
growth	NN	O	O
whose	NN	O	O
function	NN	O	O
is	NN	O	O
compromised	NN	O	O
in	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
either	NN	O	O
by	NN	O	O
direct	NN	O	O
mutation	NN	O	O
or	NN	O	O
alterations	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O
.	NN	O	O

Additional	NN	O	O
case	NN	O	O
of	NN	O	O
female	NN	O	O
monozygotic	NN	O	O
twins	NN	O	O
discordant	NN	O	O
for	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
opposite	NN	O	O
X	NN	O	O
-	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
.	NN	O	O

A	NN	O	O
pair	NN	O	O
of	NN	O	O
female	NN	O	O
monozygotic	NN	O	O
(	NN	O	O
MZ	NN	O	O
)	NN	O	O
twins	NN	O	O
,	NN	O	O
heterozygous	NN	O	O
carriers	NN	O	O
for	NN	O	O
a	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
discordant	NN	O	O
for	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
molecular	NN	O	O
studies	NN	O	O
,	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
and	NN	O	O
methylation	NN	O	O
pattern	NN	O	O
of	NN	O	O
X	NN	O	O
chromosomes	NN	O	O
to	NN	O	O
search	NN	O	O
for	NN	O	O
opposite	NN	O	O
X	NN	O	O
inactivation	NN	O	O
as	NN	O	O
an	NN	O	O
explanation	NN	O	O
of	NN	O	O
their	NN	O	O
clinical	NN	O	O
discordance	NN	O	O
.	NN	O	O

Results	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
skin	NN	O	O
fibroblast	NN	O	O
cell	NN	O	O
lines	NN	O	O
suggest	NN	O	O
a	NN	O	O
partial	NN	O	O
mirror	NN	O	O
inactivation	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
X	NN	O	O
chromosome	NN	O	O
preferentially	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
unaffected	NN	O	O
twin	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
maternal	NN	O	O
deleted	NN	O	O
X	NN	O	O
chromosome	NN	O	O
preferentially	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
twin	NN	O	O
.	NN	O	O

A	NN	O	O
review	NN	O	O
shows	NN	O	O
that	NN	O	O
MZ	NN	O	O
female	NN	O	O
twins	NN	O	O
discordant	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
diseases	NN	O	I-Disease
are	NN	O	O
not	NN	O	O
uncommon	NN	O	O
.	NN	O	O

Twinning	NN	O	O
and	NN	O	O
X	NN	O	O
inactivation	NN	O	O
may	NN	O	O
be	NN	O	O
interrelated	NN	O	O
and	NN	O	O
could	NN	O	O
explain	NN	O	O
the	NN	O	O
female	NN	O	O
twins	NN	O	O
discordant	NN	O	O
for	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
traits	NN	O	O
.	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
:	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
in	NN	O	O
6	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
presenting	NN	O	O
with	NN	O	O
5	NN	O	O
different	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Fragments	NN	O	O
of	NN	O	O
the	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
cDNA	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
adolescent	NN	O	B-Disease
ALD	NN	O	I-Disease
were	NN	O	O
amplified	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
subcloned	NN	O	O
.	NN	O	O

Bidirectional	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
disclosed	NN	O	O
a	NN	O	O
cytosine	NN	O	O
to	NN	O	O
guanine	NN	O	O
transversion	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
1451	NN	O	O
in	NN	O	O
exon	NN	O	O
five	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
substitution	NN	O	O
of	NN	O	O
proline	NN	O	O
484	NN	O	O
by	NN	O	O
arginine	NN	O	O
.	NN	O	O

Five	NN	O	O
of	NN	O	O
nine	NN	O	O
siblings	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
comprising	NN	O	O
two	NN	O	O
cerebral	NN	O	B-Disease
ALD	NN	O	I-Disease
,	NN	O	O
one	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
,	NN	O	O
one	NN	O	O
Addison	NN	O	B-Disease
only	NN	O	I-Disease
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
symptomatic	NN	O	O
mother	NN	O	O
(	NN	O	O
all	NN	O	O
accumulating	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
)	NN	O	O
carried	NN	O	O
this	NN	O	O
mutation	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
unaffected	NN	O	O
persons	NN	O	O
,	NN	O	O
in	NN	O	O
five	NN	O	O
unrelated	NN	O	O
ALD	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
twenty	NN	O	O
controls	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
this	NN	O	O
missense	NN	O	O
mutation	NN	O	O
generated	NN	O	O
the	NN	O	O
disease	NN	O	O
per	NN	O	O
se	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
metabolic	NN	O	O
defect	NN	O	O
;	NN	O	O
the	NN	O	O
different	NN	O	O
phenotypes	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
must	NN	O	O
have	NN	O	O
originated	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
additional	NN	O	O
pathogenetic	NN	O	O
factors	NN	O	O
.	NN	O	O
.	NN	O	O

Novel	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
initiation	NN	O	O
codon	NN	O	O
in	NN	O	O
the	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
in	NN	O	O
two	NN	O	O
Japanese	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
new	NN	O	O
mutation	NN	O	O
of	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
ND	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
in	NN	O	O
two	NN	O	O
Japanese	NN	O	O
males	NN	O	O
from	NN	O	O
unrelated	NN	O	O
families	NN	O	O
;	NN	O	O
they	NN	O	O
showed	NN	O	O
typical	NN	O	O
ocular	NN	O	O
features	NN	O	O
of	NN	O	O
ND	NN	O	B-Disease
but	NN	O	O
no	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
or	NN	O	O
hearing	NN	O	B-Disease
impairment	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
both	NN	O	O
patients	NN	O	O
at	NN	O	O
the	NN	O	O
initiation	NN	O	O
codon	NN	O	O
of	NN	O	O
exon	NN	O	O
2	NN	O	O
of	NN	O	O
the	NN	O	O
ND	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
ATG	NN	O	O
to	NN	O	O
GTG	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
otherwise	NN	O	O
normal	NN	O	O
nucleotide	NN	O	O
sequences	NN	O	O
.	NN	O	O

Their	NN	O	O
mothers	NN	O	O
had	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
mutant	NN	O	O
types	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
expected	NN	O	O
for	NN	O	O
heterozygotes	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
initiation	NN	O	O
codon	NN	O	O
would	NN	O	O
cause	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
ND	NN	O	B-Disease
gene	NN	O	O
expression	NN	O	O
or	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
translation	NN	O	O
thereby	NN	O	O
truncating	NN	O	O
the	NN	O	O
amino	NN	O	O
terminus	NN	O	O
of	NN	O	O
ND	NN	O	B-Disease
protein	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
rarity	NN	O	O
and	NN	O	O
marked	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ND	NN	O	B-Disease
gene	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
apparently	NN	O	O
unrelated	NN	O	O
Japanese	NN	O	O
families	NN	O	O
who	NN	O	O
have	NN	O	O
lived	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
area	NN	O	O
for	NN	O	O
over	NN	O	O
two	NN	O	O
centuries	NN	O	O
presumably	NN	O	O
share	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O
.	NN	O	O

Anticipation	NN	O	O
resulting	NN	O	O
in	NN	O	O
elimination	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
gene	NN	O	O
:	NN	O	O
a	NN	O	O
follow	NN	O	O
up	NN	O	O
study	NN	O	O
of	NN	O	O
one	NN	O	O
extended	NN	O	O
family	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
re	NN	O	O
-	NN	O	O
examined	NN	O	O
an	NN	O	O
extended	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
family	NN	O	O
,	NN	O	O
previously	NN	O	O
described	NN	O	O
in	NN	O	O
1955	NN	O	O
,	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
long	NN	O	O
term	NN	O	O
effects	NN	O	O
of	NN	O	O
anticipation	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
and	NN	O	O
in	NN	O	O
particular	NN	O	O
the	NN	O	O
implications	NN	O	O
for	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

This	NN	O	O
follow	NN	O	O
up	NN	O	O
study	NN	O	O
provides	NN	O	O
data	NN	O	O
on	NN	O	O
35	NN	O	O
gene	NN	O	O
carriers	NN	O	O
and	NN	O	O
46	NN	O	O
asymptomatic	NN	O	O
at	NN	O	O
risk	NN	O	O
family	NN	O	O
members	NN	O	O
in	NN	O	O
five	NN	O	O
generations	NN	O	O
.	NN	O	O

Clinical	NN	O	O
anticipation	NN	O	O
,	NN	O	O
defined	NN	O	O
as	NN	O	O
the	NN	O	O
cascade	NN	O	O
of	NN	O	O
mild	NN	O	O
,	NN	O	O
adult	NN	O	O
,	NN	O	O
childhood	NN	O	O
,	NN	O	O
or	NN	O	O
congenital	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
subsequent	NN	O	O
generations	NN	O	O
,	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
relentless	NN	O	O
process	NN	O	O
,	NN	O	O
occurring	NN	O	O
in	NN	O	O
all	NN	O	O
affected	NN	O	O
branches	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
.	NN	O	O

The	NN	O	O
cascade	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
proceed	NN	O	O
asynchronously	NN	O	O
in	NN	O	O
the	NN	O	O
different	NN	O	O
branches	NN	O	O
,	NN	O	O
mainly	NN	O	O
because	NN	O	O
of	NN	O	O
an	NN	O	O
unequal	NN	O	O
number	NN	O	O
of	NN	O	O
generations	NN	O	O
with	NN	O	O
mild	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
transition	NN	O	O
from	NN	O	O
the	NN	O	O
mild	NN	O	O
to	NN	O	O
the	NN	O	O
adult	NN	O	O
type	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
transmission	NN	O	O
through	NN	O	O
a	NN	O	O
male	NN	O	O
parent	NN	O	O
.	NN	O	O

Stable	NN	O	O
transmission	NN	O	O
of	NN	O	O
the	NN	O	O
asymptomatic	NN	O	O
/	NN	O	O
mild	NN	O	O
phenotype	NN	O	O
showed	NN	O	O
a	NN	O	O
female	NN	O	O
transmission	NN	O	O
bias	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
examined	NN	O	O
the	NN	O	O
extent	NN	O	O
and	NN	O	O
causes	NN	O	O
of	NN	O	O
gene	NN	O	O
loss	NN	O	O
in	NN	O	O
this	NN	O	O
pedigree	NN	O	O
.	NN	O	O

Gene	NN	O	O
loss	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
group	NN	O	O
was	NN	O	O
complete	NN	O	O
,	NN	O	O
owing	NN	O	O
to	NN	O	O
infertility	NN	O	B-Disease
of	NN	O	O
the	NN	O	O
male	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
onset	NN	O	O
disease	NN	O	O
and	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
mentally	NN	O	B-Disease
retarded	NN	O	I-Disease
patients	NN	O	O
did	NN	O	O
not	NN	O	O
procreate	NN	O	O
.	NN	O	O

Out	NN	O	O
of	NN	O	O
the	NN	O	O
46	NN	O	O
at	NN	O	O
risk	NN	O	O
subjects	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
youngest	NN	O	O
generations	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
full	NN	O	O
mutation	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
only	NN	O	O
subject	NN	O	O
who	NN	O	O
may	NN	O	O
transmit	NN	O	O
the	NN	O	O
gene	NN	O	O
to	NN	O	O
the	NN	O	O
sixth	NN	O	O
generation	NN	O	O
.	NN	O	O

No	NN	O	O
protomutation	NN	O	O
carriers	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
fourth	NN	O	O
and	NN	O	O
fifth	NN	O	O
generations	NN	O	O
.	NN	O	O

Therefore	NN	O	O
it	NN	O	O
is	NN	O	O
highly	NN	O	O
probable	NN	O	O
that	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
gene	NN	O	O
will	NN	O	O
be	NN	O	O
eliminated	NN	O	O
from	NN	O	O
this	NN	O	O
pedigree	NN	O	O
within	NN	O	O
one	NN	O	O
generation	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
population	NN	O	O
frequency	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
can	NN	O	O
at	NN	O	O
present	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
asymptomatic	NN	O	O
cases	NN	O	O
in	NN	O	O
the	NN	O	O
younger	NN	O	O
generations	NN	O	O
of	NN	O	O
known	NN	O	O
families	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
probably	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
events	NN	O	O
in	NN	O	O
the	NN	O	O
ancestral	NN	O	O
generations	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
for	NN	O	O
spinal	NN	O	B-Disease
cerebellar	NN	O	I-Disease
ataxia	NN	O	I-Disease
3	NN	O	O
(	NN	O	O
SCA3	NN	O	O
)	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
approximately	NN	O	O
3	NN	O	O
cM	NN	O	O
on	NN	O	O
chromosome	NN	O	O
14q24	NN	O	O
.	NN	O	O
3	NN	O	O
-	NN	O	O
q32	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O

SCA3	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
spinal	NN	O	B-Disease
cerebellar	NN	O	I-Disease
ataxia	NN	O	I-Disease
3	NN	O	O
,	NN	O	O
was	NN	O	O
recently	NN	O	O
mapped	NN	O	O
to	NN	O	O
a	NN	O	O
15	NN	O	O
-	NN	O	O
cM	NN	O	O
interval	NN	O	O
between	NN	O	O
D14S67	NN	O	O
and	NN	O	O
D14S81	NN	O	O
on	NN	O	O
chromosome	NN	O	O
14q	NN	O	O
,	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
of	NN	O	O
French	NN	O	O
ancestry	NN	O	O
.	NN	O	O

The	NN	O	O
SCA3	NN	O	O
candidate	NN	O	O
region	NN	O	O
has	NN	O	O
now	NN	O	O
been	NN	O	O
refined	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
with	NN	O	O
four	NN	O	O
new	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
(	NN	O	O
D14S256	NN	O	O
,	NN	O	O
D14S291	NN	O	O
,	NN	O	O
D14S280	NN	O	O
,	NN	O	O
and	NN	O	O
AFM343vf1	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
two	NN	O	O
families	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
19	NN	O	O
additional	NN	O	O
individuals	NN	O	O
were	NN	O	O
genotyped	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
third	NN	O	O
French	NN	O	O
family	NN	O	O
.	NN	O	O

Combined	NN	O	O
two	NN	O	O
-	NN	O	O
point	NN	O	O
linkage	NN	O	O
analyses	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
new	NN	O	O
markers	NN	O	O
,	NN	O	O
D14S280	NN	O	O
and	NN	O	O
AFM343vf1	NN	O	O
,	NN	O	O
are	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
SCA3	NN	O	O
locus	NN	O	O
,	NN	O	O
with	NN	O	O
maximal	NN	O	O
lod	NN	O	O
scores	NN	O	O
,	NN	O	O
at	NN	O	O
recombination	NN	O	O
fraction	NN	O	O
,	NN	O	O
(	NN	O	O
theta	NN	O	O
)	NN	O	O
=	NN	O	O
.	NN	O	O

00	NN	O	O
,	NN	O	O
of	NN	O	O
7	NN	O	O
.	NN	O	O

05	NN	O	O
and	NN	O	O
13	NN	O	O
.	NN	O	O

70	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Combined	NN	O	O
multipoint	NN	O	O
and	NN	O	O
recombinant	NN	O	O
haplotype	NN	O	O
analyses	NN	O	O
localize	NN	O	O
the	NN	O	O
SCA3	NN	O	O
locus	NN	O	O
to	NN	O	O
a	NN	O	O
3	NN	O	O
-	NN	O	O
cM	NN	O	O
interval	NN	O	O
flanked	NN	O	O
by	NN	O	O
D14S291	NN	O	O
and	NN	O	O
D14S81	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
allele	NN	O	O
for	NN	O	O
D14S280	NN	O	O
segregates	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
three	NN	O	O
kindreds	NN	O	O
.	NN	O	O

This	NN	O	O
allele	NN	O	O
is	NN	O	O
frequent	NN	O	O
in	NN	O	O
the	NN	O	O
French	NN	O	O
population	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
and	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
is	NN	O	O
not	NN	O	O
clearly	NN	O	O
established	NN	O	O
.	NN	O	O

The	NN	O	O
SCA3	NN	O	O
locus	NN	O	O
remains	NN	O	O
within	NN	O	O
the	NN	O	O
29	NN	O	O
-	NN	O	O
cM	NN	O	O
region	NN	O	O
on	NN	O	O
14q24	NN	O	O
.	NN	O	O

3	NN	O	O
-	NN	O	O
q32	NN	O	O
.	NN	O	O

2	NN	O	O
containing	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
is	NN	O	O
clinically	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
phenotype	NN	O	O
determined	NN	O	O
by	NN	O	O
SCA3	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
cannot	NN	O	O
yet	NN	O	O
be	NN	O	O
concluded	NN	O	O
that	NN	O	O
both	NN	O	O
diseases	NN	O	O
result	NN	O	O
from	NN	O	O
alterations	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O

An	NN	O	O
evaluation	NN	O	O
of	NN	O	O
genetic	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
145	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

Breast	NN	O	B-Disease
Cancer	NN	O	I-Disease
Linkage	NN	O	O
Consortium	NN	O	O
.	NN	O	O

The	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovary	NN	O	I-Disease
cancer	NN	O	I-Disease
-	NN	O	I-Disease
family	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
dominant	NN	O	O
predisposition	NN	O	O
to	NN	O	O
cancer	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovaries	NN	O	I-Disease
which	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
region	NN	O	O
17q12	NN	O	O
-	NN	O	O
q21	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
all	NN	O	O
,	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovary	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
show	NN	O	O
linkage	NN	O	O
to	NN	O	O
this	NN	O	O
susceptibility	NN	O	O
locus	NN	O	O
,	NN	O	O
designated	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
a	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
of	NN	O	O
145	NN	O	O
families	NN	O	O
with	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
families	NN	O	O
contain	NN	O	O
either	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
three	NN	O	O
or	NN	O	O
more	NN	O	O
cases	NN	O	O
of	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
(	NN	O	O
before	NN	O	O
age	NN	O	O
60	NN	O	O
years	NN	O	O
)	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
or	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

All	NN	O	O
families	NN	O	O
contained	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
case	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
an	NN	O	O
estimated	NN	O	O
76	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
145	NN	O	O
families	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
locus	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
13	NN	O	O
families	NN	O	O
with	NN	O	O
cases	NN	O	O
of	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
appear	NN	O	O
to	NN	O	O
be	NN	O	O
linked	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
estimated	NN	O	O
that	NN	O	O
92	NN	O	O
%	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
confidence	NN	O	O
interval	NN	O	O
76	NN	O	O
%	NN	O	O
-	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
no	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
and	NN	O	O
with	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
are	NN	O	O
linked	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
-	NN	O	I-Disease
family	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
genetically	NN	O	O
heterogeneous	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
large	NN	O	O
majority	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
and	NN	O	O
with	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
cases	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
essential	NN	O	B-Disease
fructosuria	NN	O	I-Disease
:	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
and	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	O
ketohexokinase	NN	O	O
(	NN	O	O
fructokinase	NN	O	O
)	NN	O	O
.	NN	O	O

Essential	NN	O	B-Disease
fructosuria	NN	O	I-Disease
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
oldest	NN	O	O
known	NN	O	O
inborn	NN	O	B-Disease
errors	NN	O	I-Disease
of	NN	O	I-Disease
metabolism	NN	O	I-Disease
.	NN	O	O

It	NN	O	O
is	NN	O	O
a	NN	O	O
benign	NN	O	O
condition	NN	O	O
which	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
result	NN	O	O
from	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
hepatic	NN	O	I-Disease
fructokinase	NN	O	I-Disease
(	NN	O	O
ketohexokinase	NN	O	O
,	NN	O	O
KHK	NN	O	O
,	NN	O	O
E	NN	O	O
.	NN	O	O
C	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O
7	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
enzyme	NN	O	O
catalyses	NN	O	O
the	NN	O	O
first	NN	O	O
step	NN	O	O
of	NN	O	O
metabolism	NN	O	O
of	NN	O	O
dietary	NN	O	O
fructose	NN	O	O
,	NN	O	O
conversion	NN	O	O
of	NN	O	O
fructose	NN	O	O
to	NN	O	O
fructose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
early	NN	O	O
recognition	NN	O	O
of	NN	O	O
this	NN	O	O
disorder	NN	O	O
,	NN	O	O
the	NN	O	O
primary	NN	O	O
structure	NN	O	O
of	NN	O	O
human	NN	O	O
KHK	NN	O	O
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
essential	NN	O	B-Disease
fructosuria	NN	O	I-Disease
have	NN	O	O
not	NN	O	O
been	NN	O	O
previously	NN	O	O
defined	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
sequencing	NN	O	O
of	NN	O	O
full	NN	O	O
-	NN	O	O
length	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
encoding	NN	O	O
human	NN	O	O
ketohexokinase	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

Alternative	NN	O	O
mRNA	NN	O	O
species	NN	O	O
and	NN	O	O
alternative	NN	O	O
KHK	NN	O	O
isozymes	NN	O	O
are	NN	O	O
produced	NN	O	O
by	NN	O	O
alternative	NN	O	O
polyadenylation	NN	O	O
and	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
KHK	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
KHK	NN	O	O
proteins	NN	O	O
show	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
sequence	NN	O	O
conservation	NN	O	O
relative	NN	O	O
to	NN	O	O
rat	NN	O	O
KHK	NN	O	O
.	NN	O	O

Direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
KHK	NN	O	O
structural	NN	O	O
gene	NN	O	O
is	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
essential	NN	O	B-Disease
fructosuria	NN	O	I-Disease
was	NN	O	O
also	NN	O	O
obtained	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
well	NN	O	O
-	NN	O	O
characterized	NN	O	O
family	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
three	NN	O	O
of	NN	O	O
eight	NN	O	O
siblings	NN	O	O
have	NN	O	O
fructosuria	NN	O	B-Disease
,	NN	O	O
all	NN	O	O
affected	NN	O	O
individuals	NN	O	O
are	NN	O	O
compound	NN	O	O
heterozygotes	NN	O	O
for	NN	O	O
two	NN	O	O
mutations	NN	O	O
Gly40Arg	NN	O	O
and	NN	O	O
Ala43Thr	NN	O	O
.	NN	O	O

Both	NN	O	O
mutations	NN	O	O
result	NN	O	O
from	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transitions	NN	O	O
,	NN	O	O
and	NN	O	O
each	NN	O	O
alters	NN	O	O
the	NN	O	O
same	NN	O	O
conserved	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
KHK	NN	O	O
protein	NN	O	O
.	NN	O	O

Neither	NN	O	O
mutation	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
a	NN	O	O
sample	NN	O	O
of	NN	O	O
52	NN	O	O
unrelated	NN	O	O
control	NN	O	O
individuals	NN	O	O
.	NN	O	O

An	NN	O	O
additional	NN	O	O
conservative	NN	O	O
amino	NN	O	O
acid	NN	O	O
change	NN	O	O
(	NN	O	O
Val49IIe	NN	O	O
)	NN	O	O
was	NN	O	O
present	NN	O	O
on	NN	O	O
the	NN	O	O
KHK	NN	O	O
allele	NN	O	O
bearing	NN	O	O
Ala43Thr	NN	O	O

Homozygous	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
crossover	NN	O	O
(	NN	O	O
fusion	NN	O	O
gene	NN	O	O
)	NN	O	O
mutation	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
Gaucher	NN	O	I-Disease
disease	NN	O	I-Disease
fetus	NN	O	O
:	NN	O	O
is	NN	O	O
this	NN	O	O
analogous	NN	O	O
to	NN	O	O
the	NN	O	O
Gaucher	NN	O	O
knock	NN	O	O
-	NN	O	O
out	NN	O	O
mouse	NN	O	O
model	NN	O	O
?	NN	O	O

Gaucher	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
GD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
beta	NN	O	I-Disease
-	NN	O	I-Disease
glucocerebrosidase	NN	O	I-Disease
(	NN	O	O
EC	NN	O	O
3	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O
2	NN	O	O
.	NN	O	O
45	NN	O	O
,	NN	O	O
gene	NN	O	O
symbol	NN	O	O
GBA	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
GD	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
CNS	NN	O	O
is	NN	O	O
not	NN	O	O
involved	NN	O	O
(	NN	O	O
nonneuronopathic	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
GD	NN	O	I-Disease
(	NN	O	O
acute	NN	O	O
neuronopathic	NN	O	O
)	NN	O	O
CNS	NN	O	O
involvement	NN	O	O
is	NN	O	O
early	NN	O	O
and	NN	O	O
rapidly	NN	O	O
progressive	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
III	NN	O	I-Disease
GD	NN	O	I-Disease
(	NN	O	O
subacute	NN	O	O
neuronopathic	NN	O	O
)	NN	O	O
CNS	NN	O	O
involvement	NN	O	O
occurs	NN	O	O
later	NN	O	O
and	NN	O	O
is	NN	O	O
slowly	NN	O	O
progressive	NN	O	O
.	NN	O	O

The	NN	O	O
T6433C	NN	O	O
(	NN	O	O
L444P	NN	O	O
)	NN	O	O
substitution	NN	O	O
is	NN	O	O
prevalent	NN	O	O
in	NN	O	O
type	NN	O	B-Disease
GD	NN	O	I-Disease
II	NN	O	I-Disease
.	NN	O	O

It	NN	O	O
may	NN	O	O
occur	NN	O	O
alone	NN	O	O
as	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	O
-	NN	O	O
pair	NN	O	O
mutation	NN	O	O
but	NN	O	O
often	NN	O	O
is	NN	O	O
found	NN	O	O
as	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
allele	NN	O	O
containing	NN	O	O
additional	NN	O	O
GBA	NN	O	O
nucleotide	NN	O	O
substitutions	NN	O	O
,	NN	O	O
G6468C	NN	O	O
(	NN	O	O
A456P	NN	O	O
)	NN	O	O
and	NN	O	O
G6482C	NN	O	O
(	NN	O	O
V460V	NN	O	O
)	NN	O	O
,	NN	O	O
without	NN	O	O
(	NN	O	O
recNciI	NN	O	O
)	NN	O	O
or	NN	O	O
with	NN	O	O
(	NN	O	O
recTL	NN	O	O
)	NN	O	O
G5957C	NN	O	O
(	NN	O	O
D409H	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
complex	NN	O	O
allele	NN	O	O
is	NN	O	O
presumed	NN	O	O
to	NN	O	O
have	NN	O	O
formed	NN	O	O
by	NN	O	O
recombination	NN	O	O
(	NN	O	O
crossover	NN	O	O
,	NN	O	O
fusion	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
structural	NN	O	O
gene	NN	O	O
with	NN	O	O
the	NN	O	O
pseudogene	NN	O	O
,	NN	O	O
which	NN	O	O
contains	NN	O	O
the	NN	O	O
mutated	NN	O	O
sequences	NN	O	O
.	NN	O	O

Two	NN	O	O
complex	NN	O	O
alleles	NN	O	O
have	NN	O	O
never	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
coexist	NN	O	O
in	NN	O	O
any	NN	O	O
individual	NN	O	O
.	NN	O	O

We	NN	O	O
devised	NN	O	O
a	NN	O	O
selective	NN	O	O
PCR	NN	O	O
method	NN	O	O
for	NN	O	O
the	NN	O	O
specific	NN	O	O
amplification	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
fusion	NN	O	O
gene	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
procedure	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
fusion	NN	O	O
gene	NN	O	O
in	NN	O	O
homozygous	NN	O	O
form	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
Macedonian	NN	O	O
/	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
GD	NN	O	B-Disease
type	NN	O	I-Disease
II	NN	O	I-Disease
fetus	NN	O	O
.	NN	O	O

Both	NN	O	O
parents	NN	O	O
were	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
recombination	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
direct	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
.	NN	O	O

A	NN	O	O
previous	NN	O	O
conceptus	NN	O	O
in	NN	O	O
this	NN	O	O
family	NN	O	O
was	NN	O	O
stillborn	NN	O	B-Disease
at	NN	O	O
36	NN	O	O
weeks	NN	O	O
,	NN	O	O
with	NN	O	O
features	NN	O	O
of	NN	O	O
severe	NN	O	O
type	NN	O	B-Disease
II	NN	O	I-Disease
GD	NN	O	I-Disease
.	NN	O	O

Neonates	NN	O	O
showing	NN	O	O
a	NN	O	O
severe	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
,	NN	O	O
analogous	NN	O	O
to	NN	O	O
the	NN	O	O
early	NN	O	O
neonatal	NN	O	O
lethal	NN	O	O
disease	NN	O	O
occurring	NN	O	O
in	NN	O	O
mice	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
null	NN	O	O
allele	NN	O	O
produced	NN	O	O
by	NN	O	O
targeted	NN	O	O
disruption	NN	O	O
of	NN	O	O
GBA	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
elsewhere	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
specific	NN	O	O
mutations	NN	O	O
in	NN	O	O
these	NN	O	O
cases	NN	O	O
have	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
characterized	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

Late	NN	O	O
infantile	NN	O	O
metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
in	NN	O	O
Israel	NN	O	O
.	NN	O	O

Metachromatic	NN	O	B-Disease
Leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
neurodegenerative	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
which	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
enzyme	NN	O	O
,	NN	O	O
Aryl	NN	O	O
sulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
is	NN	O	O
deficient	NN	O	O
.	NN	O	O

The	NN	O	O
disease	NN	O	O
is	NN	O	O
inherited	NN	O	O
as	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
trait	NN	O	O
and	NN	O	O
its	NN	O	O
frequency	NN	O	O
is	NN	O	O
estimated	NN	O	O
to	NN	O	O
be	NN	O	O
1	NN	O	O
/	NN	O	O
40	NN	O	O
,	NN	O	O
000	NN	O	O
live	NN	O	O
births	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
of	NN	O	O
ARSA	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
up	NN	O	O
to	NN	O	O
now	NN	O	O
eight	NN	O	O
mutations	NN	O	O
causing	NN	O	O
MLD	NN	O	B-Disease
have	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

Another	NN	O	O
mutation	NN	O	O
,	NN	O	O
PD	NN	O	O
,	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
deficiency	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
in	NN	O	O
vitro	NN	O	O
(	NN	O	O
pseudodeficiency	NN	O	O
)	NN	O	O
without	NN	O	O
any	NN	O	O
known	NN	O	O
clinical	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
PD	NN	O	O
mutation	NN	O	O
is	NN	O	O
frequent	NN	O	O
in	NN	O	O
all	NN	O	O
populations	NN	O	O
.	NN	O	O

In	NN	O	O
Israel	NN	O	O
,	NN	O	O
late	NN	O	O
infantile	NN	O	O
MLD	NN	O	B-Disease
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
very	NN	O	O
frequent	NN	O	O
in	NN	O	O
a	NN	O	O
small	NN	O	O
Jewish	NN	O	O
isolate	NN	O	O
,	NN	O	O
the	NN	O	O
Habbanite	NN	O	O
Jews	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
75	NN	O	O
live	NN	O	O
births	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
Habbanite	NN	O	O
population	NN	O	O
,	NN	O	O
the	NN	O	O
mutation	NN	O	O
occurred	NN	O	O
on	NN	O	O
an	NN	O	O
allele	NN	O	O
with	NN	O	O
the	NN	O	O
PD	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
loss	NN	O	O
of	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
C	NN	O	O
>	NN	O	O
T	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
change	NN	O	O
of	NN	O	O
proline	NN	O	O
to	NN	O	O
leucine	NN	O	O
.	NN	O	O

MLD	NN	O	B-Disease
is	NN	O	O
also	NN	O	O
frequent	NN	O	O
among	NN	O	O
Moslem	NN	O	O
Arabs	NN	O	O
in	NN	O	O
Jerusalem	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
is	NN	O	O
a	NN	O	O
transition	NN	O	O
G	NN	O	O
>	NN	O	O
A	NN	O	O
destroying	NN	O	O
the	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
exon	NN	O	O
2	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
late	NN	O	O
infantile	NN	O	O
MLD	NN	O	B-Disease
from	NN	O	O
different	NN	O	O
ethnic	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
Christian	NN	O	O
Arabs	NN	O	O
in	NN	O	O
Israel	NN	O	O
also	NN	O	O
have	NN	O	O
a	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
(	NN	O	O
1	NN	O	O
/	NN	O	O
10	NN	O	O
,	NN	O	O
000	NN	O	O
live	NN	O	O
births	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
is	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

Knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
mutations	NN	O	O
causing	NN	O	O
MLD	NN	O	B-Disease
in	NN	O	O
these	NN	O	O
defined	NN	O	O
populations	NN	O	O
will	NN	O	O
allow	NN	O	O
a	NN	O	O
carrier	NN	O	O
screening	NN	O	O
program	NN	O	O
to	NN	O	O
be	NN	O	O
carried	NN	O	O
out	NN	O	O
and	NN	O	O
prevent	NN	O	O
the	NN	O	O
birth	NN	O	O
of	NN	O	O
additional	NN	O	O
affected	NN	O	O
children	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
(	NN	O	O
G103D	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
triple	NN	O	O
helix	NN	O	O
produces	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
moderately	NN	O	O
severe	NN	O	O
chondrodysplasia	NN	O	B-Disease
phenotype	NN	O	O
that	NN	O	O
results	NN	O	O
from	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
,	NN	O	O
COL2A1	NN	O	O
.	NN	O	O

Characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
disorder	NN	O	O
include	NN	O	O
a	NN	O	O
short	NN	O	B-Disease
trunk	NN	O	I-Disease
and	NN	O	I-Disease
extremities	NN	O	I-Disease
,	NN	O	O
mid	NN	O	B-Disease
-	NN	O	I-Disease
face	NN	O	I-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
cleft	NN	O	B-Disease
palate	NN	O	I-Disease
,	NN	O	O
myopia	NN	O	B-Disease
,	NN	O	O
retinal	NN	O	B-Disease
detachment	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
hearing	NN	O	B-Disease
loss	NN	O	I-Disease
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
deletions	NN	O	O
of	NN	O	O
all	NN	O	O
or	NN	O	O
part	NN	O	O
of	NN	O	O
exon	NN	O	O
12	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
mutations	NN	O	O
within	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
may	NN	O	O
primarily	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
phenotype	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
SSCP	NN	O	O
to	NN	O	O
analyze	NN	O	O
an	NN	O	O
amplified	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
containing	NN	O	O
exon	NN	O	O
12	NN	O	O
from	NN	O	O
seven	NN	O	O
individuals	NN	O	O
with	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
.	NN	O	O

An	NN	O	O
abnormality	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
one	NN	O	O
patient	NN	O	O
.	NN	O	O

DNA	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
patient	NN	O	O
was	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
transition	NN	O	O
that	NN	O	O
implied	NN	O	O
substitution	NN	O	O
of	NN	O	O
glycine103	NN	O	O
of	NN	O	O
the	NN	O	O
triple	NN	O	O
helical	NN	O	O
domain	NN	O	O
by	NN	O	O
aspartate	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
DNA	NN	O	O
from	NN	O	O
either	NN	O	O
of	NN	O	O
the	NN	O	O
clinically	NN	O	O
unaffected	NN	O	O
parents	NN	O	O
of	NN	O	O
the	NN	O	O
proband	NN	O	O
.	NN	O	O

Protein	NN	O	O
microsequencing	NN	O	O
demonstrated	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
abnormal	NN	O	O
allele	NN	O	O
in	NN	O	O
cartilage	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
point	NN	O	O
mutations	NN	O	O
which	NN	O	O
result	NN	O	O
in	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
can	NN	O	O
produce	NN	O	O
Kniest	NN	O	B-Disease
dysplasia	NN	O	I-Disease
and	NN	O	O
further	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
alteration	NN	O	O
of	NN	O	O
a	NN	O	O
domain	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
the	NN	O	O
region	NN	O	O
encoded	NN	O	O
by	NN	O	O
exon	NN	O	O
12	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
type	NN	O	O
II	NN	O	O
collagen	NN	O	O
protein	NN	O	O
leads	NN	O	O
to	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O
.	NN	O	O

CAG	NN	O	O
expansions	NN	O	O
in	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
for	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
at	NN	O	O
chromosome	NN	O	O
14q32	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
containing	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
and	NN	O	O
mapped	NN	O	O
it	NN	O	O
to	NN	O	O
chromosome	NN	O	O
14q32	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
the	NN	O	O
genetic	NN	O	O
locus	NN	O	O
for	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
normal	NN	O	O
individuals	NN	O	O
the	NN	O	O
gene	NN	O	O
contains	NN	O	O
between	NN	O	O
13	NN	O	O
and	NN	O	O
36	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
,	NN	O	O
whereas	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
clinically	NN	O	O
diagnosed	NN	O	O
patients	NN	O	O
and	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
pathological	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
MJD	NN	O	B-Disease
show	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
-	NN	O	O
number	NN	O	O
(	NN	O	O
from	NN	O	O
68	NN	O	O
-	NN	O	O
79	NN	O	O
)	NN	O	O
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
and	NN	O	O
genomic	NN	O	O
cloning	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
related	NN	O	O
genes	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
raise	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
similar	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
related	NN	O	O
genes	NN	O	O
may	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
diseases	NN	O	O
similar	NN	O	O
to	NN	O	O
MJD	NN	O	B-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
50	NN	O	O
probands	NN	O	O
with	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
/	NN	O	I-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
for	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
candidate	NN	O	O
gene	NN	O	O
,	NN	O	O
using	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
on	NN	O	O
PCR	NN	O	O
-	NN	O	O
amplified	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
eight	NN	O	O
putative	NN	O	O
disease	NN	O	O
-	NN	O	O
causing	NN	O	O
alterations	NN	O	O
were	NN	O	O
identified	NN	O	O
four	NN	O	O
of	NN	O	O
these	NN	O	O
are	NN	O	O
frameshifts	NN	O	O
and	NN	O	O
two	NN	O	O
are	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
two	NN	O	O
missense	NN	O	O
mutations	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
changes	NN	O	O
the	NN	O	O
final	NN	O	O
cysteine	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
zinc	NN	O	O
finger	NN	O	O
motif	NN	O	O
to	NN	O	O
glycine	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
model	NN	O	O
,	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
this	NN	O	O
candidate	NN	O	O
gene	NN	O	O
is	NN	O	O
in	NN	O	O
fact	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

The	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
mutations	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
large	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
indicates	NN	O	O
that	NN	O	O
clinical	NN	O	O
application	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutation	NN	O	O
testing	NN	O	O
will	NN	O	O
be	NN	O	O
technically	NN	O	O
challenging	NN	O	O
.	NN	O	O
.	NN	O	O

Confirmation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
linked	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
in	NN	O	O
ten	NN	O	O
families	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
genetic	NN	O	O
evidence	NN	O	O
supporting	NN	O	O
the	NN	O	O
identity	NN	O	O
of	NN	O	O
the	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
through	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
germline	NN	O	O
mutations	NN	O	O
in	NN	O	O
63	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
and	NN	O	O
10	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
in	NN	O	O
ten	NN	O	O
families	NN	O	O
with	NN	O	O
cancer	NN	O	B-Disease
linked	NN	O	O
to	NN	O	O
chromosome	NN	O	O
17q21	NN	O	O
.	NN	O	O

Nine	NN	O	O
different	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
screening	NN	O	O
BRCA1	NN	O	O
DNA	NN	O	O
and	NN	O	O
RNA	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Seven	NN	O	O
mutations	NN	O	O
lead	NN	O	O
to	NN	O	O
protein	NN	O	O
truncations	NN	O	O
at	NN	O	O
sites	NN	O	O
throughout	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

One	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
which	NN	O	O
occurred	NN	O	O
independently	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
a	NN	O	O
cysteine	NN	O	O
in	NN	O	O
the	NN	O	O
zinc	NN	O	O
binding	NN	O	O
domain	NN	O	O
.	NN	O	O

An	NN	O	O
intronic	NN	O	O
single	NN	O	O
basepair	NN	O	O
substitution	NN	O	O
destroys	NN	O	O
an	NN	O	O
acceptor	NN	O	O
site	NN	O	O
and	NN	O	O
activates	NN	O	O
a	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
site	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
59	NN	O	O
basepair	NN	O	O
insertion	NN	O	O
and	NN	O	O
chain	NN	O	O
termination	NN	O	O
.	NN	O	O

The	NN	O	O
four	NN	O	O
families	NN	O	O
with	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
had	NN	O	O
chain	NN	O	O
termination	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

High	NN	O	O
resolution	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
suggests	NN	O	O
one	NN	O	O
ancestral	NN	O	O
predisposing	NN	O	O
haplotype	NN	O	O
for	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
causing	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
amplification	NN	O	O
of	NN	O	O
an	NN	O	O
unstable	NN	O	O
trinucleotide	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
in	NN	O	O
over	NN	O	O
99	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
global	NN	O	O
DM	NN	O	B-Disease
population	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
in	NN	O	O
complete	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
with	NN	O	O
an	NN	O	O
Alu	NN	O	O
element	NN	O	O
polymorphism	NN	O	O
within	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
few	NN	O	O
ancestral	NN	O	O
mutations	NN	O	O
.	NN	O	O

A	NN	O	O
recent	NN	O	O
analysis	NN	O	O
utilizing	NN	O	O
this	NN	O	O
polymorphism	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
flanking	NN	O	O
dinucleotide	NN	O	O
marker	NN	O	O
,	NN	O	O
suggested	NN	O	O
that	NN	O	O
similar	NN	O	O
to	NN	O	O
Fragile	NN	O	B-Disease
X	NN	O	I-Disease
syndrome	NN	O	I-Disease
,	NN	O	O
DM	NN	O	B-Disease
exhibited	NN	O	O
a	NN	O	O
founder	NN	O	O
effect	NN	O	O
(	NN	O	O
Imbert	NN	O	O
et	NN	O	O
al	NN	O	O
.	NN	O	O
,	NN	O	O
1993	NN	O	O
Nature	NN	O	O
Genet	NN	O	O
.	NN	O	O
4	NN	O	O
,	NN	O	O
72	NN	O	O
-	NN	O	O
76	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
low	NN	O	O
reproductive	NN	O	O
fitness	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
congenital	NN	O	O
DM	NN	O	B-Disease
(	NN	O	O
the	NN	O	O
endpoint	NN	O	O
of	NN	O	O
genetic	NN	O	O
anticipation	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
)	NN	O	O
suggests	NN	O	O
a	NN	O	O
higher	NN	O	O
rate	NN	O	O
of	NN	O	O
new	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
high	NN	O	O
resolution	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
locus	NN	O	O
using	NN	O	O
PCR	NN	O	O
based	NN	O	O
assays	NN	O	O
of	NN	O	O
nine	NN	O	O
polymorphisms	NN	O	O
,	NN	O	O
spanning	NN	O	O
a	NN	O	O
physical	NN	O	O
distance	NN	O	O
of	NN	O	O
30	NN	O	O
kb	NN	O	O
,	NN	O	O
within	NN	O	O
and	NN	O	O
immediately	NN	O	O
flanking	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
persistent	NN	O	O
complete	NN	O	O
allelic	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
with	NN	O	O
all	NN	O	O
these	NN	O	O
polymorphisms	NN	O	O
provides	NN	O	O
further	NN	O	O
support	NN	O	O
to	NN	O	O
previous	NN	O	O
observations	NN	O	O
and	NN	O	O
suggests	NN	O	O
more	NN	O	O
strongly	NN	O	O
that	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
mutation	NN	O	O
occurred	NN	O	O
on	NN	O	O
the	NN	O	O
background	NN	O	O
of	NN	O	O
a	NN	O	O
particular	NN	O	O
haplotype	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
became	NN	O	O
inherently	NN	O	O
unstable	NN	O	O
and	NN	O	O
therefore	NN	O	O
predisposed	NN	O	O
to	NN	O	O
amplification	NN	O	O
.	NN	O	O

Genetic	NN	O	O
instability	NN	O	O
in	NN	O	O
human	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
microsatellites	NN	O	O
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
and	NN	O	O
found	NN	O	O
that	NN	O	O
5	NN	O	O
out	NN	O	O
of	NN	O	O
10	NN	O	O
of	NN	O	O
the	NN	O	O
ovarian	NN	O	B-Disease
tumor	NN	O	I-Disease
cell	NN	O	O
lines	NN	O	O
are	NN	O	O
genetically	NN	O	O
unstable	NN	O	O
at	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
loci	NN	O	O
analyzed	NN	O	O
.	NN	O	O

In	NN	O	O
clones	NN	O	O
and	NN	O	O
subclones	NN	O	O
derived	NN	O	O
serially	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
(	NN	O	O
2774	NN	O	O
;	NN	O	O
serous	NN	O	B-Disease
cystadenocarcinoma	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
very	NN	O	O
high	NN	O	O
proportion	NN	O	O
of	NN	O	O
microsatellites	NN	O	O
distributed	NN	O	O
in	NN	O	O
many	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
genome	NN	O	O
change	NN	O	O
their	NN	O	O
size	NN	O	O
in	NN	O	O
a	NN	O	O
mercurial	NN	O	O
fashion	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
genomic	NN	O	O
instability	NN	O	O
in	NN	O	O
ovarian	NN	O	B-Disease
tumors	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
dynamic	NN	O	O
and	NN	O	O
ongoing	NN	O	O
process	NN	O	O
whose	NN	O	O
high	NN	O	O
frequency	NN	O	O
may	NN	O	O
have	NN	O	O
been	NN	O	O
previously	NN	O	O
underestimated	NN	O	O
by	NN	O	O
PCR	NN	O	O
-	NN	O	O
based	NN	O	O
allelotyping	NN	O	O
of	NN	O	O
bulk	NN	O	O
tumor	NN	O	B-Disease
tissue	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
instability	NN	O	O
in	NN	O	O
one	NN	O	O
ovarian	NN	O	B-Disease
tumor	NN	O	I-Disease
as	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
(	NN	O	O
R524P	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
mismatch	NN	O	O
-	NN	O	O
repair	NN	O	O
gene	NN	O	O
MSH2	NN	O	O
(	NN	O	O
Salmonella	NN	O	O
MutS	NN	O	O
homologue	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
hereditary	NN	O	B-Disease
nonpolyposis	NN	O	I-Disease
colorectal	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Patient	NN	O	O
2774	NN	O	O
was	NN	O	O
a	NN	O	O
38	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
heterozygote	NN	O	O
,	NN	O	O
and	NN	O	O
her	NN	O	O
normal	NN	O	O
tissue	NN	O	O
carried	NN	O	O
both	NN	O	O
mutant	NN	O	O
and	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
MSH2	NN	O	O
gene	NN	O	O
.	NN	O	O

However	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
allele	NN	O	O
was	NN	O	O
lost	NN	O	O
at	NN	O	O
some	NN	O	O
point	NN	O	O
early	NN	O	O
during	NN	O	O
tumorigenesis	NN	O	O
so	NN	O	O
that	NN	O	O
DNA	NN	O	O
isolated	NN	O	O
either	NN	O	O
from	NN	O	O
the	NN	O	O
patients	NN	O	O
ovarian	NN	O	B-Disease
tumor	NN	O	I-Disease
or	NN	O	O
from	NN	O	O
the	NN	O	O
2774	NN	O	O
cell	NN	O	O
line	NN	O	O
carries	NN	O	O
only	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
MSH2	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
genetic	NN	O	O
instability	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
and	NN	O	O
cell	NN	O	O
line	NN	O	O
DNA	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
mismatch	NN	O	O
-	NN	O	O
repair	NN	O	O
gene	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
MSH2	NN	O	O
gene	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
onset	NN	O	O
and	NN	O	O
/	NN	O	O
or	NN	O	O
progression	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
primary	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
carcinomas	NN	O	I-Disease
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
data	NN	O	O
from	NN	O	O
familial	NN	O	B-Disease
tumors	NN	O	I-Disease
suggest	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
confers	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
ovarian	NN	O	B-Disease
and	NN	O	I-Disease
early	NN	O	I-Disease
-	NN	O	I-Disease
onset	NN	O	I-Disease
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
region	NN	O	O
is	NN	O	O
also	NN	O	O
subject	NN	O	O
to	NN	O	O
allelic	NN	O	O
loss	NN	O	O
in	NN	O	O
sporadic	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
,	NN	O	O
an	NN	O	O
indication	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
may	NN	O	O
occur	NN	O	O
somatically	NN	O	O
in	NN	O	O
these	NN	O	O
tumors	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
coding	NN	O	O
region	NN	O	O
was	NN	O	O
examined	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
primary	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
tumors	NN	O	I-Disease
that	NN	O	O
show	NN	O	O
allele	NN	O	O
loss	NN	O	O
at	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
locus	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
3	NN	O	O
of	NN	O	O
32	NN	O	O
breast	NN	O	O
and	NN	O	O
1	NN	O	O
of	NN	O	O
12	NN	O	O
ovarian	NN	O	B-Disease
carcinomas	NN	O	I-Disease
;	NN	O	O
all	NN	O	O
four	NN	O	O
mutations	NN	O	O
were	NN	O	O
germline	NN	O	O
alterations	NN	O	O
and	NN	O	O
occurred	NN	O	O
in	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
cancers	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
critical	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancers	NN	O	I-Disease
that	NN	O	O
arise	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	O
germline	NN	O	O
allele	NN	O	O
.	NN	O	O
.	NN	O	O

PAX6	NN	O	O
gene	NN	O	O
dosage	NN	O	O
effect	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
cataracts	NN	O	I-Disease
,	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
anophthalmia	NN	O	B-Disease
and	NN	O	O
central	NN	O	B-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
defects	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
human	NN	O	O
eye	NN	O	O
malformation	NN	O	O
aniridia	NN	O	B-Disease
results	NN	O	O
from	NN	O	O
haploinsufficiency	NN	O	B-Disease
of	NN	O	I-Disease
PAX6	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
paired	NN	O	O
box	NN	O	O
DNA	NN	O	O
-	NN	O	O
binding	NN	O	O
protein	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
this	NN	O	O
dosage	NN	O	O
effect	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
two	NN	O	O
PAX6	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
family	NN	O	O
segregating	NN	O	O
aniridia	NN	O	B-Disease
and	NN	O	O
a	NN	O	O
milder	NN	O	O
syndrome	NN	O	O
consisting	NN	O	O
of	NN	O	O
congenital	NN	O	B-Disease
cataracts	NN	O	I-Disease
and	NN	O	O
late	NN	O	O
onset	NN	O	O
corneal	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
at	NN	O	O
codons	NN	O	O
103	NN	O	O
and	NN	O	O
353	NN	O	O
,	NN	O	O
truncate	NN	O	O
PAX6	NN	O	O
within	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
paired	NN	O	O
and	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
PST	NN	O	O
domains	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
PST	NN	O	O
domain	NN	O	O
activates	NN	O	O
transcription	NN	O	O
autonomously	NN	O	O
and	NN	O	O
the	NN	O	O
mutant	NN	O	O
form	NN	O	O
has	NN	O	O
partial	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
compound	NN	O	O
heterozygote	NN	O	O
had	NN	O	O
severe	NN	O	O
craniofacial	NN	O	B-Disease
and	NN	O	I-Disease
central	NN	O	I-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
defects	NN	O	I-Disease
and	NN	O	O
no	NN	O	B-Disease
eyes	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
malformations	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
homozygous	NN	O	O
Sey	NN	O	O
mice	NN	O	O
and	NN	O	O
suggests	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
PAX6	NN	O	O
in	NN	O	O
controlling	NN	O	O
the	NN	O	O
migration	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
specific	NN	O	O
neuronal	NN	O	O
progenitor	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
physical	NN	O	O
map	NN	O	O
and	NN	O	O
candidate	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
-	NN	O	O
21	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
constructed	NN	O	O
a	NN	O	O
physical	NN	O	O
map	NN	O	O
of	NN	O	O
a	NN	O	O
4	NN	O	O
cM	NN	O	O
region	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
-	NN	O	O
21	NN	O	O
that	NN	O	O
contains	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
gene	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

The	NN	O	O
map	NN	O	O
comprises	NN	O	O
a	NN	O	O
contig	NN	O	O
of	NN	O	O
137	NN	O	O
overlapping	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosomes	NN	O	O
and	NN	O	O
P1	NN	O	O
clones	NN	O	O
,	NN	O	O
onto	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
placed	NN	O	O
112	NN	O	O
PCR	NN	O	O
markers	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
localized	NN	O	O
more	NN	O	O
than	NN	O	O
20	NN	O	O
genes	NN	O	O
on	NN	O	O
this	NN	O	O
map	NN	O	O
,	NN	O	O
ten	NN	O	O
of	NN	O	O
which	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
previously	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
isolated	NN	O	O
30	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
representing	NN	O	O
partial	NN	O	O
sequences	NN	O	O
of	NN	O	O
as	NN	O	O
yet	NN	O	O
unidentified	NN	O	O
genes	NN	O	O
.	NN	O	O

Two	NN	O	O
genes	NN	O	O
that	NN	O	O
lie	NN	O	O
within	NN	O	O
a	NN	O	O
narrow	NN	O	O
region	NN	O	O
defined	NN	O	O
by	NN	O	O
meiotic	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
patients	NN	O	O
have	NN	O	O
been	NN	O	O
sequenced	NN	O	O
in	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
patients	NN	O	O
without	NN	O	O
revealing	NN	O	O
any	NN	O	O
deleterious	NN	O	O
mutations	NN	O	O
.	NN	O	O

These	NN	O	O
new	NN	O	O
reagents	NN	O	O
should	NN	O	O
facilitate	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
LEC	NN	O	O
rat	NN	O	O
has	NN	O	O
a	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
copper	NN	O	O
transporting	NN	O	O
ATPase	NN	O	O
gene	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
Long	NN	O	O
-	NN	O	O
Evans	NN	O	O
Cinnamon	NN	O	O
(	NN	O	O
LEC	NN	O	O
)	NN	O	O
rat	NN	O	O
shows	NN	O	O
similarity	NN	O	O
to	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
in	NN	O	O
many	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
features	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
cDNAs	NN	O	O
for	NN	O	O
the	NN	O	O
rat	NN	O	O
gene	NN	O	O
(	NN	O	O
Atp7b	NN	O	O
)	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
ATP7B	NN	O	O
)	NN	O	O
and	NN	O	O
have	NN	O	O
used	NN	O	O
them	NN	O	O
to	NN	O	O
identify	NN	O	O
a	NN	O	O
partial	NN	O	O
deletion	NN	O	O
in	NN	O	O
the	NN	O	O
Atp7b	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
LEC	NN	O	O
rat	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
removes	NN	O	O
at	NN	O	O
least	NN	O	O
900	NN	O	O
bp	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
at	NN	O	O
the	NN	O	O
3	NN	O	O
end	NN	O	O
,	NN	O	O
includes	NN	O	O
the	NN	O	O
crucial	NN	O	O
ATP	NN	O	O
binding	NN	O	O
domain	NN	O	O
and	NN	O	O
extends	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
convincing	NN	O	O
evidence	NN	O	O
for	NN	O	O
defining	NN	O	O
the	NN	O	O
LEC	NN	O	O
rat	NN	O	O
as	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
model	NN	O	O
will	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
studying	NN	O	O
liver	NN	O	O
pathophysiology	NN	O	O
,	NN	O	O
for	NN	O	O
developing	NN	O	O
therapy	NN	O	O
for	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
pathway	NN	O	O
of	NN	O	O
copper	NN	O	O
transport	NN	O	O
and	NN	O	O
its	NN	O	O
possible	NN	O	O
interaction	NN	O	O
with	NN	O	O
other	NN	O	O
heavy	NN	O	O
metals	NN	O	O
.	NN	O	O
.	NN	O	O

Genomic	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
peroxisomal	NN	O	B-Disease
disorder	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
a	NN	O	O
severe	NN	O	O
neurodegenerative	NN	O	B-Disease
disease	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
an	NN	O	O
impairment	NN	O	B-Disease
of	NN	O	I-Disease
very	NN	O	I-Disease
long	NN	O	I-Disease
chain	NN	O	I-Disease
fatty	NN	O	I-Disease
acids	NN	O	I-Disease
beta	NN	O	I-Disease
-	NN	O	I-Disease
oxidation	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
identified	NN	O	O
by	NN	O	O
positional	NN	O	O
cloning	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
ALD	NN	O	B-Disease
,	NN	O	O
located	NN	O	O
in	NN	O	O
Xq28	NN	O	O
.	NN	O	O

It	NN	O	O
encodes	NN	O	O
a	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
"	NN	O	O
ABC	NN	O	O
"	NN	O	O
superfamily	NN	O	O
of	NN	O	O
membrane	NN	O	O
-	NN	O	O
associated	NN	O	O
transporters	NN	O	O
that	NN	O	O
shows	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
significant	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
70	NN	O	O
-	NN	O	O
kDa	NN	O	O
peroxisomal	NN	O	O
membrane	NN	O	O
protein	NN	O	O
(	NN	O	O
PMP70	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
a	NN	O	O
detailed	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
structure	NN	O	O
.	NN	O	O

It	NN	O	O
extends	NN	O	O
over	NN	O	O
21	NN	O	O
kb	NN	O	O
and	NN	O	O
consists	NN	O	O
of	NN	O	O
10	NN	O	O
exons	NN	O	O
.	NN	O	O

To	NN	O	O
facilitate	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
ALD	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
intronic	NN	O	O
sequences	NN	O	O
flanking	NN	O	O
the	NN	O	O
exons	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	O
5	NN	O	O
promoter	NN	O	O
region	NN	O	O
.	NN	O	O

Sequences	NN	O	O
present	NN	O	O
in	NN	O	O
distal	NN	O	O
exons	NN	O	O
cross	NN	O	O
-	NN	O	O
hybridize	NN	O	O
strongly	NN	O	O
to	NN	O	O
additional	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
genome	NN	O	O
.	NN	O	O

The	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
has	NN	O	O
been	NN	O	O
positioned	NN	O	O
on	NN	O	O
a	NN	O	O
pulsed	NN	O	O
-	NN	O	O
field	NN	O	O
map	NN	O	O
between	NN	O	O
DXS15	NN	O	O
and	NN	O	O
the	NN	O	O
L1CAM	NN	O	O
gene	NN	O	O
,	NN	O	O
about	NN	O	O
650	NN	O	O
kb	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
color	NN	O	O
pigment	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
frequent	NN	O	O
occurrence	NN	O	O
of	NN	O	O
color	NN	O	O
vision	NN	O	O
anomalies	NN	O	O
observed	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
the	NN	O	O
adult	NN	O	O
onset	NN	O	O
form	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
thus	NN	O	O
does	NN	O	O
not	NN	O	O
represent	NN	O	O
a	NN	O	O
contiguous	NN	O	B-Disease
gene	NN	O	I-Disease
syndrome	NN	O	I-Disease
but	NN	O	O
a	NN	O	O
secondary	NN	O	O
manifestation	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

The	NN	O	O
murine	NN	O	O
homologues	NN	O	O
of	NN	O	O
the	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
Hdh	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
alpha	NN	O	O
-	NN	O	O
adducin	NN	O	O
gene	NN	O	O
(	NN	O	O
Add1	NN	O	O
)	NN	O	O
map	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
5	NN	O	O
within	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
conserved	NN	O	O
synteny	NN	O	O
with	NN	O	O
human	NN	O	O
chromosome	NN	O	O
4p16	NN	O	O
.	NN	O	O
3	NN	O	O
.	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
severe	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
neurodegenerative	NN	O	I-Disease
disorder	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
(	NN	O	O
IT15	NN	O	O
)	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
reported	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
Hdh	NN	O	O
,	NN	O	O
the	NN	O	O
murine	NN	O	O
homologue	NN	O	O
of	NN	O	O
IT15	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
using	NN	O	O
an	NN	O	O
interspecific	NN	O	O
backcross	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
mapped	NN	O	O
both	NN	O	O
Hdh	NN	O	O
and	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
human	NN	O	O
alpha	NN	O	O
-	NN	O	O
adducin	NN	O	O
(	NN	O	O
Add1	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
membrane	NN	O	O
-	NN	O	O
associated	NN	O	O
cytoskeletal	NN	O	O
protein	NN	O	O
gene	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
map	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
position	NN	O	O
on	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
5	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
associated	NN	O	O
with	NN	O	O
ancestral	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
and	NN	O	O
show	NN	O	O
no	NN	O	O
recombination	NN	O	O
with	NN	O	O
D5H4S43	NN	O	O
,	NN	O	O
D5H4S115	NN	O	O
,	NN	O	O
and	NN	O	O
D5H4S62	NN	O	O
,	NN	O	O
the	NN	O	O
murine	NN	O	O
homologues	NN	O	O
of	NN	O	O
D4S43	NN	O	O
,	NN	O	O
D4S115	NN	O	O
,	NN	O	O
and	NN	O	O
D4S62	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Further	NN	O	O
mapping	NN	O	O
studies	NN	O	O
of	NN	O	O
humans	NN	O	O
,	NN	O	O
mice	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
mammalian	NN	O	O
species	NN	O	O
should	NN	O	O
reveal	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
rearrangements	NN	O	O
affecting	NN	O	O
this	NN	O	O
chromosomal	NN	O	O
segment	NN	O	O
during	NN	O	O
mammalian	NN	O	O
evolution	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
cholesteryl	NN	O	B-Disease
ester	NN	O	I-Disease
transfer	NN	O	I-Disease
protein	NN	O	I-Disease
deficiency	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
two	NN	O	O
prevalent	NN	O	O
mutations	NN	O	O
as	NN	O	O
a	NN	O	O
major	NN	O	O
determinant	NN	O	O
of	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
cholesterol	NN	O	O
.	NN	O	O

Genetic	NN	O	O
determinants	NN	O	O
of	NN	O	O
HDL	NN	O	O
cholesterol	NN	O	O
(	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
)	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
described	NN	O	O
plasma	NN	O	O
cholesteryl	NN	O	B-Disease
ester	NN	O	I-Disease
transfer	NN	O	I-Disease
protein	NN	O	I-Disease
(	NN	O	I-Disease
CETP	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
an	NN	O	O
intron	NN	O	O
14	NN	O	O
G	NN	O	O
(	NN	O	O
+	NN	O	O
1	NN	O	O
)	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
A	NN	O	O
mutation	NN	O	O
(	NN	O	O
Int14	NN	O	O
A	NN	O	O
)	NN	O	O
in	NN	O	O
several	NN	O	O
families	NN	O	O
with	NN	O	O
very	NN	O	O
high	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
levels	NN	O	O
in	NN	O	O
Japan	NN	O	O
.	NN	O	O

Subjects	NN	O	O
with	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
100	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
130	NN	O	O
)	NN	O	O
were	NN	O	O
screened	NN	O	O
by	NN	O	O
PCR	NN	O	O
single	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
CETP	NN	O	O
gene	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
or	NN	O	O
primer	NN	O	O
-	NN	O	O
mediated	NN	O	O
restriction	NN	O	O
map	NN	O	O
modification	NN	O	O
of	NN	O	O
PCR	NN	O	O
products	NN	O	O
a	NN	O	O
novel	NN	O	O
intron	NN	O	O
14	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
mutation	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
T	NN	O	O
insertion	NN	O	O
at	NN	O	O
position	NN	O	O
+	NN	O	O
3	NN	O	O
from	NN	O	O
the	NN	O	O
exon14	NN	O	O
/	NN	O	O
intron14	NN	O	O
boundary	NN	O	O
(	NN	O	O
Int14	NN	O	O
T	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
Asp442	NN	O	O
to	NN	O	O
Gly	NN	O	O
)	NN	O	O
within	NN	O	O
exon	NN	O	O
15	NN	O	O
(	NN	O	O
D442G	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
Int14	NN	O	O
T	NN	O	O
mutation	NN	O	O
was	NN	O	O
only	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
D442G	NN	O	O
and	NN	O	O
Int14	NN	O	O
A	NN	O	O
mutations	NN	O	O
were	NN	O	O
highly	NN	O	O
prevalent	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
60	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
,	NN	O	O
with	NN	O	O
combined	NN	O	O
allele	NN	O	O
frequencies	NN	O	O
of	NN	O	O
9	NN	O	O
%	NN	O	O
,	NN	O	O
12	NN	O	O
%	NN	O	O
,	NN	O	O
21	NN	O	O
%	NN	O	O
and	NN	O	O
43	NN	O	O
%	NN	O	O
for	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
60	NN	O	O
-	NN	O	O
79	NN	O	O
,	NN	O	O
80	NN	O	O
-	NN	O	O
99	NN	O	O
,	NN	O	O
100	NN	O	O
-	NN	O	O
119	NN	O	O
,	NN	O	O
and	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
120	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
prevalences	NN	O	O
of	NN	O	O
the	NN	O	O
D442G	NN	O	O
and	NN	O	O
Int14	NN	O	O
A	NN	O	O
mutations	NN	O	O
were	NN	O	O
extremely	NN	O	O
high	NN	O	O
in	NN	O	O
a	NN	O	O
general	NN	O	O
sample	NN	O	O
of	NN	O	O
Japanese	NN	O	O
men	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
236	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
heterozygote	NN	O	O
frequencies	NN	O	O
of	NN	O	O
7	NN	O	O
%	NN	O	O
and	NN	O	O
2	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
two	NN	O	O
mutations	NN	O	O
accounted	NN	O	O
for	NN	O	O
about	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
variance	NN	O	O
of	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
phenotype	NN	O	O
in	NN	O	O
a	NN	O	O
genetic	NN	O	O
compound	NN	O	O
heterozygote	NN	O	O
(	NN	O	O
Int14	NN	O	O
T	NN	O	O
and	NN	O	O
Int14	NN	O	O
A	NN	O	O
)	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
Int14	NN	O	O
A	NN	O	O
homozygotes	NN	O	O
(	NN	O	O
no	NN	O	O
detectable	NN	O	O
CETP	NN	O	O
and	NN	O	O
markedly	NN	O	O
increased	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
Int14	NN	O	O
T	NN	O	O
produces	NN	O	O
a	NN	O	O
null	NN	O	O
allele	NN	O	O
.	NN	O	O

In	NN	O	O
four	NN	O	O
D442G	NN	O	O
homozygotes	NN	O	O
,	NN	O	O
mean	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
levels	NN	O	O
(	NN	O	O
86	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
26	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
)	NN	O	O
were	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
Int14	NN	O	O
A	NN	O	O
homozygotes	NN	O	O
(	NN	O	O
158	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
35	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
)	NN	O	O
,	NN	O	O
reflecting	NN	O	O
residual	NN	O	O
CETP	NN	O	O
activity	NN	O	O
in	NN	O	O
plasma	NN	O	O
.	NN	O	O

In	NN	O	O
47	NN	O	O
D442G	NN	O	O
heterozygotes	NN	O	O
,	NN	O	O
mean	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
levels	NN	O	O
were	NN	O	O
91	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
23	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
level	NN	O	O
in	NN	O	O
D442G	NN	O	O
homozygotes	NN	O	O
,	NN	O	O
and	NN	O	O
significantly	NN	O	O
greater	NN	O	O
than	NN	O	O
mean	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
levels	NN	O	O
in	NN	O	O
Int14	NN	O	O
A	NN	O	O
heterozygotes	NN	O	O
(	NN	O	O
69	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
15	NN	O	O
mg	NN	O	O
/	NN	O	O
dl	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
D442G	NN	O	O
mutation	NN	O	O
acts	NN	O	O
differently	NN	O	O
to	NN	O	O
the	NN	O	O
null	NN	O	O
mutations	NN	O	O
with	NN	O	O
weaker	NN	O	O
effects	NN	O	O
on	NN	O	O
HDL	NN	O	O
in	NN	O	O
the	NN	O	O
homozygous	NN	O	O
state	NN	O	O
and	NN	O	O
stronger	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
heterozygotes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
dominant	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
partially	NN	O	O
defective	NN	O	O
allele	NN	O	O
.	NN	O	O

CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
reflecting	NN	O	O
two	NN	O	O
prevalent	NN	O	O
mutations	NN	O	O
(	NN	O	O
D442G	NN	O	O
and	NN	O	O
Int14	NN	O	O
A	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
genetic	NN	O	B-Disease
deficiency	NN	O	I-Disease
state	NN	O	O
which	NN	O	O
is	NN	O	O
sufficiently	NN	O	O
common	NN	O	O
to	NN	O	O
explain	NN	O	O
a	NN	O	O
significant	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
variation	NN	O	O
in	NN	O	O
HDL	NN	O	O
-	NN	O	O
C	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
:	NN	O	O
effects	NN	O	O
of	NN	O	O
chenodeoxycholic	NN	O	O
acid	NN	O	O
,	NN	O	O
pravastatin	NN	O	O
,	NN	O	O
and	NN	O	O
combined	NN	O	O
use	NN	O	O
.	NN	O	O

Treatments	NN	O	O
by	NN	O	O
oral	NN	O	O
administration	NN	O	O
of	NN	O	O
chenodeoxycholic	NN	O	O
acid	NN	O	O
(	NN	O	O
CDCA	NN	O	O
)	NN	O	O
alone	NN	O	O
,	NN	O	O
3	NN	O	O
-	NN	O	O
hydroxy	NN	O	O
-	NN	O	O
3	NN	O	O
-	NN	O	O
methylglutaryl	NN	O	O
(	NN	O	O
HMG	NN	O	O
)	NN	O	O
CoA	NN	O	O
reductase	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
pravastatin	NN	O	O
)	NN	O	O
alone	NN	O	O
,	NN	O	O
and	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
drugs	NN	O	O
were	NN	O	O
attempted	NN	O	O
for	NN	O	O
7	NN	O	O
patients	NN	O	O
with	NN	O	O
cerebrotendinous	NN	O	B-Disease
xanthomatosis	NN	O	I-Disease
(	NN	O	O
CTX	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

CDCA	NN	O	O
treatment	NN	O	O
at	NN	O	O
a	NN	O	O
dose	NN	O	O
of	NN	O	O
300	NN	O	O
mg	NN	O	O
/	NN	O	O
day	NN	O	O
reduced	NN	O	O
serum	NN	O	O
cholestanol	NN	O	O
(	NN	O	O
67	NN	O	O
.	NN	O	O
3	NN	O	O
%	NN	O	O
reduction	NN	O	O
)	NN	O	O
,	NN	O	O
lathosterol	NN	O	O
(	NN	O	O
50	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
campesterol	NN	O	O
(	NN	O	O
61	NN	O	O
.	NN	O	O
7	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
sitosterol	NN	O	O
(	NN	O	O
12	NN	O	O
.	NN	O	O
7	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
sera	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
changed	NN	O	O
to	NN	O	O
be	NN	O	O
"	NN	O	O
atherogenic	NN	O	O
"	NN	O	O
;	NN	O	O
total	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
triglyceride	NN	O	O
and	NN	O	O
low	NN	O	O
-	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
LDL	NN	O	O
)	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
were	NN	O	O
increased	NN	O	O
,	NN	O	O
while	NN	O	O
high	NN	O	O
-	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
-	NN	O	O
cholesterol	NN	O	O
was	NN	O	O
decreased	NN	O	O
.	NN	O	O

Contrarily	NN	O	O
,	NN	O	O
pravastatin	NN	O	O
at	NN	O	O
a	NN	O	O
dose	NN	O	O
of	NN	O	O
10	NN	O	O
mg	NN	O	O
/	NN	O	O
day	NN	O	O
improved	NN	O	O
the	NN	O	O
sera	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
to	NN	O	O
be	NN	O	O
markedly	NN	O	O
"	NN	O	O
anti	NN	O	O
-	NN	O	O
atherogenic	NN	O	O
"	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
reductions	NN	O	O
of	NN	O	O
cholestanol	NN	O	O
(	NN	O	O
30	NN	O	O
.	NN	O	O
4	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
lathosterol	NN	O	O
(	NN	O	O
44	NN	O	O
.	NN	O	O
0	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
campesterol	NN	O	O
(	NN	O	O
22	NN	O	O
.	NN	O	O
9	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
sitosterol	NN	O	O
(	NN	O	O
9	NN	O	O
.	NN	O	O
6	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
inadequate	NN	O	O
.	NN	O	O

Combined	NN	O	O
treatment	NN	O	O
with	NN	O	O
CDCA	NN	O	O
and	NN	O	O
pravastatin	NN	O	O
showed	NN	O	O
good	NN	O	O
overlapping	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
each	NN	O	O
drug	NN	O	O
alone	NN	O	O
.	NN	O	O

The	NN	O	O
sera	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
apparently	NN	O	O
more	NN	O	O
"	NN	O	O
anti	NN	O	O
-	NN	O	O
atherogenic	NN	O	O
"	NN	O	O
than	NN	O	O
those	NN	O	O
after	NN	O	O
CDCA	NN	O	O
treatment	NN	O	O
.	NN	O	O

Serum	NN	O	O
cholestanol	NN	O	O
concentration	NN	O	O
was	NN	O	O
still	NN	O	O
2	NN	O	O
.	NN	O	O

7	NN	O	O
times	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
controls	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
serum	NN	O	O
lathosterol	NN	O	O
level	NN	O	O
was	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
enhancement	NN	O	O
of	NN	O	O
overall	NN	O	O
cholesterol	NN	O	O
synthesis	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
sufficiently	NN	O	O
suppressed	NN	O	O
.	NN	O	O

Plant	NN	O	O
sterol	NN	O	O
levels	NN	O	O
were	NN	O	O
also	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
.	NN	O	O

The	NN	O	O
combination	NN	O	O
of	NN	O	O
CDCA	NN	O	O
and	NN	O	O
pravastatin	NN	O	O
was	NN	O	O
a	NN	O	O
good	NN	O	O
treatment	NN	O	O
for	NN	O	O
CTX	NN	O	B-Disease
,	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
improvement	NN	O	O
of	NN	O	O
serum	NN	O	O
lipoprotein	NN	O	O
metabolism	NN	O	O
,	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
cholesterol	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
reductions	NN	O	O
of	NN	O	O
cholestanol	NN	O	O
and	NN	O	O
plant	NN	O	O
sterol	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
of	NN	O	O
7	NN	O	O
patients	NN	O	O
,	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
disease	NN	O	O
was	NN	O	O
arrested	NN	O	O
,	NN	O	O
but	NN	O	O
dramatic	NN	O	O
effects	NN	O	O
on	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
,	NN	O	O
xanthoma	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
electrophysiological	NN	O	O
findings	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
found	NN	O	O
after	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
these	NN	O	O
drugs	NN	O	O

Mutation	NN	O	O
spectrum	NN	O	O
in	NN	O	O
the	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
of	NN	O	O
Danish	NN	O	O
and	NN	O	O
Swedish	NN	O	O
choroideremia	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

The	NN	O	O
recent	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
complete	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
of	NN	O	O
the	NN	O	O
choroideremia	NN	O	B-Disease
(	NN	O	O
CHM	NN	O	B-Disease
)	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
exon	NN	O	O
-	NN	O	O
intron	NN	O	O
boundaries	NN	O	O
has	NN	O	O
paved	NN	O	O
the	NN	O	O
way	NN	O	O
to	NN	O	O
mutation	NN	O	O
detection	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
classical	NN	O	O
choroideremia	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
performed	NN	O	O
mutation	NN	O	O
screening	NN	O	O
in	NN	O	O
patients	NN	O	O
from	NN	O	O
15	NN	O	O
Danish	NN	O	O
and	NN	O	O
Swedish	NN	O	O
families	NN	O	O
by	NN	O	O
using	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
and	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
PCR	NN	O	O
-	NN	O	O
SSCP	NN	O	O
)	NN	O	O
technique	NN	O	O
.	NN	O	O

Causative	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
12	NN	O	O
families	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
a	NN	O	O
substantial	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
can	NN	O	O
be	NN	O	O
identified	NN	O	O
by	NN	O	O
this	NN	O	O
approach	NN	O	O
.	NN	O	O

In	NN	O	O
four	NN	O	O
of	NN	O	O
these	NN	O	O
families	NN	O	O
deletions	NN	O	O
of	NN	O	O
different	NN	O	O
sizes	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
one	NN	O	O
patient	NN	O	O
,	NN	O	O
the	NN	O	O
deletion	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
only	NN	O	O
one	NN	O	O
exon	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
another	NN	O	O
the	NN	O	O
deletion	NN	O	O
comprised	NN	O	O
the	NN	O	O
entire	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Mapping	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
endpoints	NN	O	O
in	NN	O	O
these	NN	O	O
four	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
another	NN	O	O
11	NN	O	O
male	NN	O	O
patients	NN	O	O
with	NN	O	O
sizeable	NN	O	O
deletions	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
construct	NN	O	O
a	NN	O	O
very	NN	O	O
detailed	NN	O	O
map	NN	O	O
of	NN	O	O
intervals	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
of	NN	O	O
Xq21	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
remaining	NN	O	O
11	NN	O	O
Danish	NN	O	O
and	NN	O	O
Swedish	NN	O	O
families	NN	O	O
at	NN	O	O
least	NN	O	O
8	NN	O	O
causative	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
by	NN	O	O
PCR	NN	O	O
-	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
all	NN	O	O
CHM	NN	O	B-Disease
gene	NN	O	O
mutations	NN	O	O
detected	NN	O	O
thus	NN	O	O
far	NN	O	O
in	NN	O	O
choroideremia	NN	O	B-Disease
patients	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
the	NN	O	O
introduction	NN	O	O
of	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O
.	NN	O	O

Predominance	NN	O	O
of	NN	O	O
the	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
phenotype	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
in	NN	O	O
The	NN	O	O
Netherlands	NN	O	O
:	NN	O	O
a	NN	O	O
survey	NN	O	O
of	NN	O	O
30	NN	O	O
kindreds	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
inherited	NN	O	B-Disease
disorder	NN	O	I-Disease
of	NN	O	O
peroxisomal	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
associated	NN	O	O
with	NN	O	O
accumulation	NN	O	O
of	NN	O	O
saturated	NN	O	O
very	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
central	NN	O	O
and	NN	O	O
peripheral	NN	O	O
demyelination	NN	O	B-Disease
and	NN	O	O
in	NN	O	O
impaired	NN	O	B-Disease
function	NN	O	I-Disease
of	NN	O	I-Disease
adrenal	NN	O	I-Disease
cortex	NN	O	I-Disease
and	NN	O	I-Disease
testes	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
is	NN	O	O
highly	NN	O	O
variable	NN	O	O
,	NN	O	O
childhood	NN	O	B-Disease
cerebral	NN	O	I-Disease
ALD	NN	O	I-Disease
(	NN	O	O
CCALD	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
being	NN	O	O
the	NN	O	O
main	NN	O	O
variants	NN	O	O
.	NN	O	O

We	NN	O	O
explored	NN	O	O
the	NN	O	O
30	NN	O	O
Dutch	NN	O	O
kindreds	NN	O	O
well	NN	O	O
known	NN	O	O
to	NN	O	O
the	NN	O	O
Dutch	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
/	NN	O	O
AMN	NN	O	B-Disease
Study	NN	O	O
Group	NN	O	O
and	NN	O	O
phenotyped	NN	O	O
77	NN	O	O
male	NN	O	O
patients	NN	O	O
35	NN	O	O
(	NN	O	O
46	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
AMN	NN	O	B-Disease
and	NN	O	O
24	NN	O	O
(	NN	O	O
31	NN	O	O
%	NN	O	O
)	NN	O	O
CCALD	NN	O	B-Disease
or	NN	O	O
adolescent	NN	O	B-Disease
cerebral	NN	O	I-Disease
ALD	NN	O	I-Disease
(	NN	O	O
AdolCALD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

These	NN	O	O
percentages	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
previous	NN	O	O
reports	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
25	NN	O	O
to	NN	O	O
28	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
developed	NN	O	O
AMN	NN	O	B-Disease
and	NN	O	O
53	NN	O	O
to	NN	O	O
57	NN	O	O
%	NN	O	O
CCALD	NN	O	B-Disease
or	NN	O	O
AdolCALD	NN	O	B-Disease
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
-	NN	O	O
-	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
the	NN	O	O
Netherlands	NN	O	O
-	NN	O	O
-	NN	O	O
AMN	NN	O	B-Disease
may	NN	O	O
be	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
phenotype	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
ALD	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
gene	NN	O	O
encodes	NN	O	O
an	NN	O	O
80	NN	O	O
kDa	NN	O	O
membrane	NN	O	O
protein	NN	O	O
.	NN	O	O

An	NN	O	O
antibody	NN	O	O
against	NN	O	O
the	NN	O	O
synthetic	NN	O	O
C	NN	O	O
-	NN	O	O
terminal	NN	O	O
peptides	NN	O	O
deduced	NN	O	O
from	NN	O	O
the	NN	O	O
cDNA	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
was	NN	O	O
produced	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
antibody	NN	O	O
reacted	NN	O	O
with	NN	O	O
the	NN	O	O
80	NN	O	O
kDa	NN	O	O
band	NN	O	O
protein	NN	O	O
in	NN	O	O
control	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
while	NN	O	O
no	NN	O	O
bands	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
(	NN	O	O
#	NN	O	O
163	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
mRNA	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
undetectable	NN	O	O
based	NN	O	O
on	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
293T	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
full	NN	O	O
-	NN	O	O
coding	NN	O	O
cDNA	NN	O	O
inserted	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
vector	NN	O	O
produced	NN	O	O
a	NN	O	O
new	NN	O	O
80	NN	O	O
kDa	NN	O	O
protein	NN	O	O
,	NN	O	O
as	NN	O	O
detected	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
immunocytological	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
staining	NN	O	O
was	NN	O	O
in	NN	O	O
a	NN	O	O
punctate	NN	O	O
pattern	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
fibroblasts	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
punctate	NN	O	O
staining	NN	O	O
in	NN	O	O
the	NN	O	O
#	NN	O	O
163	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
thus	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
encodes	NN	O	O
an	NN	O	O
80	NN	O	O
kDa	NN	O	O
membrane	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Isolation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
McLeod	NN	O	B-Disease
syndrome	NN	O	I-Disease
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
novel	NN	O	O
membrane	NN	O	O
transport	NN	O	O
protein	NN	O	O
.	NN	O	O

McLeod	NN	O	B-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
multisystem	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
the	NN	O	O
neuromuscular	NN	O	O
and	NN	O	O
hematopoietic	NN	O	O
systems	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
assembled	NN	O	O
a	NN	O	O
cosmid	NN	O	O
contig	NN	O	O
of	NN	O	O
360	NN	O	O
kb	NN	O	O
that	NN	O	O
encompasses	NN	O	O
the	NN	O	O
McLeod	NN	O	B-Disease
gene	NN	O	O
locus	NN	O	O
.	NN	O	O

A	NN	O	O
50	NN	O	O
kb	NN	O	O
deletion	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
screening	NN	O	O
DNA	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
radiolabeled	NN	O	O
whole	NN	O	O
cosmids	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
transcription	NN	O	O
units	NN	O	O
were	NN	O	O
identified	NN	O	O
within	NN	O	O
this	NN	O	O
deletion	NN	O	O
.	NN	O	O

The	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
them	NN	O	O
,	NN	O	O
designated	NN	O	O
as	NN	O	O
XK	NN	O	O
,	NN	O	O
correlates	NN	O	O
closely	NN	O	O
to	NN	O	O
the	NN	O	O
McLeod	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

XK	NN	O	O
encodes	NN	O	O
a	NN	O	O
novel	NN	O	O
protein	NN	O	O
with	NN	O	O
structural	NN	O	O
characteristics	NN	O	O
of	NN	O	O
prokaryotic	NN	O	O
and	NN	O	O
eukaryotic	NN	O	O
membrane	NN	O	O
transport	NN	O	O
proteins	NN	O	O
.	NN	O	O

Nucleotide	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
XK	NN	O	O
from	NN	O	O
two	NN	O	O
unrelated	NN	O	O
McLeod	NN	O	B-Disease
patients	NN	O	O
has	NN	O	O
identified	NN	O	O
point	NN	O	O
mutations	NN	O	O
at	NN	O	O
conserved	NN	O	O
splice	NN	O	O
donor	NN	O	O
and	NN	O	O
acceptor	NN	O	O
sites	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
XK	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
McLeod	NN	O	B-Disease
syndrome	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
spastic	NN	O	I-Disease
paraplegia	NN	O	I-Disease
and	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
are	NN	O	O
allelic	NN	O	B-Disease
disorders	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
locus	NN	O	O
.	NN	O	O

Three	NN	O	O
forms	NN	O	O
of	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
spastic	NN	O	I-Disease
paraplegia	NN	O	I-Disease
(	NN	O	O
SPG	NN	O	B-Disease
)	NN	O	O
have	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

One	NN	O	O
locus	NN	O	O
(	NN	O	O
SPG	NN	O	O
1	NN	O	O
)	NN	O	O
maps	NN	O	O
to	NN	O	O
Xq28	NN	O	O
while	NN	O	O
two	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
forms	NN	O	O
map	NN	O	O
to	NN	O	O
Xq22	NN	O	O
(	NN	O	O
SPG2	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
rare	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
dysmyelinating	NN	O	I-Disease
disorder	NN	O	I-Disease
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
Pelizaeus	NN	O	B-Disease
-	NN	O	I-Disease
Merzbacher	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
PMD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
Xq21	NN	O	O
-	NN	O	O
q22	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
proteolipid	NN	O	O
protein	NN	O	O
gene	NN	O	O
(	NN	O	O
PLP	NN	O	O
)	NN	O	O
which	NN	O	O
encodes	NN	O	O
two	NN	O	O
myelin	NN	O	O
proteins	NN	O	O
,	NN	O	O
PLP	NN	O	O
and	NN	O	O
DM20	NN	O	O
.	NN	O	O

While	NN	O	O
narrowing	NN	O	O
the	NN	O	O
genetic	NN	O	O
interval	NN	O	O
containing	NN	O	O
SPG2	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
pedigree	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
PLP	NN	O	O
was	NN	O	O
the	NN	O	O
closest	NN	O	O
marker	NN	O	O
to	NN	O	O
the	NN	O	O
disease	NN	O	O
locus	NN	O	O
,	NN	O	O
implicating	NN	O	O
PLP	NN	O	O
as	NN	O	O
a	NN	O	O
possible	NN	O	O
candidate	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
point	NN	O	O
mutation	NN	O	O
(	NN	O	O
His139Tyr	NN	O	O
)	NN	O	O
in	NN	O	O
exon	NN	O	O
3B	NN	O	O
of	NN	O	O
an	NN	O	O
affected	NN	O	O
male	NN	O	O
produces	NN	O	O
a	NN	O	O
mutant	NN	O	O
PLP	NN	O	O
but	NN	O	O
a	NN	O	O
normal	NN	O	O
DM20	NN	O	O
,	NN	O	O
and	NN	O	O
segregates	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
(	NN	O	O
Zmax	NN	O	O
=	NN	O	O
6	NN	O	O
.	NN	O	O
63	NN	O	O
,	NN	O	O
theta	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
00	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
that	NN	O	O
SPG2	NN	O	O
and	NN	O	O
PMD	NN	O	B-Disease
are	NN	O	O
allelic	NN	O	B-Disease
disorders	NN	O	I-Disease

Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
mutations	NN	O	O
among	NN	O	O
Jewish	NN	O	O
and	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
patients	NN	O	O
.	NN	O	O

Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
leukodystrophy	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
the	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
aspartoacylase	NN	O	I-Disease
(	NN	O	O
ASPA	NN	O	O
)	NN	O	O
.	NN	O	O

Sixty	NN	O	O
-	NN	O	O
four	NN	O	O
probands	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ASPA	NN	O	O
gene	NN	O	O
.	NN	O	O

Three	NN	O	O
point	NN	O	O
mutations	NN	O	O
-	NN	O	O
-	NN	O	O
693C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
854A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
,	NN	O	O
and	NN	O	O
914C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
693C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
and	NN	O	O
914C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
base	NN	O	O
changes	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
nonsense	NN	O	O
tyr231	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
ter	NN	O	O
and	NN	O	O
missense	NN	O	O
ala305	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
glu	NN	O	O
mutations	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
lead	NN	O	O
to	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
ASPA	NN	O	O
activity	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
expression	NN	O	O
studies	NN	O	O
.	NN	O	O

The	NN	O	O
854A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
transversion	NN	O	O
converted	NN	O	O
glu	NN	O	O
to	NN	O	O
ala	NN	O	O
in	NN	O	O
codon	NN	O	O
285	NN	O	O
.	NN	O	O

The	NN	O	O
glu285	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
ala	NN	O	O
mutant	NN	O	O
ASPA	NN	O	O
has	NN	O	O
2	NN	O	O
.	NN	O	O

5	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
expressed	NN	O	O
by	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
enzyme	NN	O	O
.	NN	O	O

A	NN	O	O
fourth	NN	O	O
mutation	NN	O	O
,	NN	O	O
433	NN	O	O
-	NN	O	O
-	NN	O	O
2	NN	O	O
(	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
transition	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
at	NN	O	O
the	NN	O	O
splice	NN	O	O
-	NN	O	O
acceptor	NN	O	O
site	NN	O	O
in	NN	O	O
intron	NN	O	O
2	NN	O	O
.	NN	O	O

The	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
would	NN	O	O
lead	NN	O	O
to	NN	O	O
skipping	NN	O	O
of	NN	O	O
exon	NN	O	O
3	NN	O	O
,	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
frameshift	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
would	NN	O	O
produce	NN	O	O
aberrant	NN	O	O
ASPA	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
128	NN	O	O
unrelated	NN	O	O
Canavan	NN	O	B-Disease
chromosomes	NN	O	O
analyzed	NN	O	O
,	NN	O	O
88	NN	O	O
were	NN	O	O
from	NN	O	O
probands	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
descent	NN	O	O
.	NN	O	O

The	NN	O	O
glu285	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
ala	NN	O	O
mutation	NN	O	O
was	NN	O	O
predominant	NN	O	O
(	NN	O	O
82	NN	O	O
.	NN	O	O
9	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
tyr231	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
ter	NN	O	O
(	NN	O	O
14	NN	O	O
.	NN	O	O
8	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
433	NN	O	O
-	NN	O	O
-	NN	O	O
2	NN	O	O
(	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
)	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
mutations	NN	O	O
account	NN	O	O
for	NN	O	O
98	NN	O	O
.	NN	O	O

8	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
Canavan	NN	O	B-Disease
chromosomes	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
origin	NN	O	O
.	NN	O	O

The	NN	O	O
ala305	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
glu	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
exclusively	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
probands	NN	O	O
of	NN	O	O
European	NN	O	O
descent	NN	O	O
and	NN	O	O
constituted	NN	O	O
60	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
40	NN	O	O
mutant	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

Predominant	NN	O	O
occurrence	NN	O	O
of	NN	O	O
certain	NN	O	O
mutations	NN	O	O
among	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
and	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
patients	NN	O	O
with	NN	O	O
Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
would	NN	O	O
suggest	NN	O	O
a	NN	O	O
founding	NN	O	O
-	NN	O	O
father	NN	O	O
effect	NN	O	O
in	NN	O	O
propagation	NN	O	O
of	NN	O	O
these	NN	O	O
mutant	NN	O	O
chromosomes	NN	O	O

Intelligence	NN	O	O
quotient	NN	O	O
profile	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
intergenerational	NN	O	O
deficit	NN	O	O
,	NN	O	O
and	NN	O	O
correlation	NN	O	O
with	NN	O	O
CTG	NN	O	O
amplification	NN	O	O
.	NN	O	O

An	NN	O	O
abbreviated	NN	O	O
Wechsler	NN	O	O
Adult	NN	O	O
Intelligence	NN	O	O
Scale	NN	O	O
Revised	NN	O	O
(	NN	O	O
WAIS	NN	O	O
-	NN	O	O
R	NN	O	O
)	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
assess	NN	O	O
verbal	NN	O	O
and	NN	O	O
arithmetical	NN	O	O
cognitive	NN	O	O
performance	NN	O	O
in	NN	O	O
55	NN	O	O
subjects	NN	O	O
with	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
covering	NN	O	O
all	NN	O	O
grades	NN	O	O
of	NN	O	O
disease	NN	O	O
severity	NN	O	O
,	NN	O	O
and	NN	O	O
31	NN	O	O
controls	NN	O	O
at	NN	O	O
50	NN	O	O
%	NN	O	O
risk	NN	O	O
of	NN	O	O
inheriting	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

Scaled	NN	O	O
scores	NN	O	O
from	NN	O	O
the	NN	O	O
assessment	NN	O	O
were	NN	O	O
converted	NN	O	O
into	NN	O	O
an	NN	O	O
intelligence	NN	O	O
quotient	NN	O	O
(	NN	O	O
IQ	NN	O	O
)	NN	O	O
estimation	NN	O	O
on	NN	O	O
each	NN	O	O
person	NN	O	O
.	NN	O	O

Significant	NN	O	O
IQ	NN	O	O
differences	NN	O	O
were	NN	O	O
found	NN	O	O
between	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
all	NN	O	O
55	NN	O	O
DM	NN	O	B-Disease
subjects	NN	O	O
(	NN	O	O
mean	NN	O	O
90	NN	O	O
.	NN	O	O
2	NN	O	O
,	NN	O	O
SD	NN	O	O
16	NN	O	O
.	NN	O	O
1	NN	O	O
)	NN	O	O
and	NN	O	O
31	NN	O	O
controls	NN	O	O
(	NN	O	O
102	NN	O	O
.	NN	O	O
6	NN	O	O
,	NN	O	O
SD	NN	O	O
9	NN	O	O
.	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
sex	NN	O	O
differences	NN	O	O
in	NN	O	O
either	NN	O	O
group	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
15	NN	O	O
affected	NN	O	O
parents	NN	O	O
(	NN	O	O
99	NN	O	O
.	NN	O	O
3	NN	O	O
,	NN	O	O
SD	NN	O	O
12	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
their	NN	O	O
affected	NN	O	O
children	NN	O	O
(	NN	O	O
88	NN	O	O
.	NN	O	O
1	NN	O	O
,	NN	O	O
SD	NN	O	O
17	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
where	NN	O	O
significance	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
parental	NN	O	O
sex	NN	O	O
being	NN	O	O
female	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
15	NN	O	O
pairs	NN	O	O
of	NN	O	O
affected	NN	O	O
sibs	NN	O	O
(	NN	O	O
89	NN	O	O
.	NN	O	O
6	NN	O	O
,	NN	O	O
SD	NN	O	O
13	NN	O	O
.	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
their	NN	O	O
normal	NN	O	O
sibs	NN	O	O
(	NN	O	O
100	NN	O	O
.	NN	O	O
2	NN	O	O
,	NN	O	O
SD	NN	O	O
7	NN	O	O
.	NN	O	O
6	NN	O	O
)	NN	O	O
.	NN	O	O

IQ	NN	O	O
steadily	NN	O	O
declined	NN	O	O
as	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
onset	NN	O	O
of	NN	O	O
signs	NN	O	O
and	NN	O	O
symptoms	NN	O	O
decreased	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
size	NN	O	O
increased	NN	O	O
.	NN	O	O

The	NN	O	O
correlation	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
linear	NN	O	O
with	NN	O	O
age	NN	O	O
of	NN	O	O
onset	NN	O	O
.	NN	O	O

The	NN	O	O
correlation	NN	O	O
of	NN	O	O
IQ	NN	O	O
difference	NN	O	O
and	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
difference	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
parent	NN	O	O
-	NN	O	O
child	NN	O	O
pairs	NN	O	O
and	NN	O	O
normal	NN	O	O
sib	NN	O	O
-	NN	O	O
affected	NN	O	O
sib	NN	O	O
pairs	NN	O	O
was	NN	O	O
poor	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
CTG	NN	O	O
expansion	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
reliable	NN	O	O
predictor	NN	O	O
of	NN	O	O
IQ	NN	O	O
either	NN	O	O
in	NN	O	O
individual	NN	O	O
persons	NN	O	O
or	NN	O	O
families	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
of	NN	O	O
cognitive	NN	O	O
function	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
required	NN	O	O
to	NN	O	O
clarify	NN	O	O
specific	NN	O	O
deficits	NN	O	O
characteristic	NN	O	O
of	NN	O	O
this	NN	O	O
patient	NN	O	O
group	NN	O	O

Adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
cytogenetic	NN	O	O
deletion	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
resulting	NN	O	O
from	NN	O	O
a	NN	O	O
maternal	NN	O	O
intrachromosomal	NN	O	O
insertion	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
laboratory	NN	O	O
findings	NN	O	O
in	NN	O	O
an	NN	O	O
institutionalised	NN	O	O
adult	NN	O	O
patient	NN	O	O
originally	NN	O	O
referred	NN	O	O
for	NN	O	O
autism	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
high	NN	O	O
risk	NN	O	O
of	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
was	NN	O	O
predicted	NN	O	O
when	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
5	NN	O	O
,	NN	O	O
del	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
(	NN	O	O
q15q22	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
her	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
MCC	NN	O	O
and	NN	O	O
APC	NN	O	B-Disease
genes	NN	O	O
confirmed	NN	O	O
by	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
.	NN	O	O

Adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
and	NN	O	O
carcinoma	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
rectum	NN	O	I-Disease
were	NN	O	O
subsequently	NN	O	O
diagnosed	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

She	NN	O	O
was	NN	O	O
profoundly	NN	O	O
mentally	NN	O	B-Disease
retarded	NN	O	I-Disease
,	NN	O	O
autistic	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
had	NN	O	O
minor	NN	O	O
dysmorphic	NN	O	B-Disease
features	NN	O	I-Disease
consistent	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
previous	NN	O	O
patients	NN	O	O
with	NN	O	O
similar	NN	O	O
deletions	NN	O	O
.	NN	O	O

The	NN	O	O
deletion	NN	O	O
arose	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
recombination	NN	O	O
within	NN	O	O
the	NN	O	O
small	NN	O	O
insertion	NN	O	O
loop	NN	O	O
formed	NN	O	O
at	NN	O	O
meiosis	NN	O	O
by	NN	O	O
the	NN	O	O
direct	NN	O	O
insertion	NN	O	O
(	NN	O	O
dir	NN	O	O
ins	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
(	NN	O	O
q22	NN	O	O
.	NN	O	O
3q14	NN	O	O
.	NN	O	O
2q15	NN	O	O
)	NN	O	O
)	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
mother	NN	O	O
.	NN	O	O

This	NN	O	O
family	NN	O	O
further	NN	O	O
confirms	NN	O	O
the	NN	O	O
cytogenetic	NN	O	O
mapping	NN	O	O
of	NN	O	O
both	NN	O	O
MCC	NN	O	O
and	NN	O	O
APC	NN	O	B-Disease
genes	NN	O	O
to	NN	O	O
5q22	NN	O	O
and	NN	O	O
comparison	NN	O	O
with	NN	O	O
other	NN	O	O
recent	NN	O	O
cases	NN	O	O
suggests	NN	O	O
that	NN	O	O
both	NN	O	O
genes	NN	O	O
and	NN	O	O
their	NN	O	O
closely	NN	O	O
linked	NN	O	O
markers	NN	O	O
lie	NN	O	O
within	NN	O	O
the	NN	O	O
5q22	NN	O	O
.	NN	O	O

1	NN	O	O
subband	NN	O	O

Familial	NN	O	B-Disease
male	NN	O	I-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
is	NN	O	O
not	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
locus	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q	NN	O	O
.	NN	O	O

Breast	NN	O	B-Disease
cancer	NN	O	I-Disease
in	NN	O	O
men	NN	O	O
is	NN	O	O
about	NN	O	O
a	NN	O	O
hundredfold	NN	O	O
less	NN	O	O
common	NN	O	O
than	NN	O	O
in	NN	O	O
women	NN	O	O
and	NN	O	O
this	NN	O	O
has	NN	O	O
hindered	NN	O	O
research	NN	O	O
into	NN	O	O
its	NN	O	O
genetic	NN	O	O
basis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
22	NN	O	O
families	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
case	NN	O	O
of	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
locus	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
strong	NN	O	O
evidence	NN	O	O
against	NN	O	O
linkage	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
(	NN	O	O
lod	NN	O	O
score	NN	O	O
-	NN	O	O
16	NN	O	O
.	NN	O	O
63	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
best	NN	O	O
estimate	NN	O	O
of	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
linked	NN	O	O
families	NN	O	O
was	NN	O	O
0	NN	O	O
%	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
CI	NN	O	O
0	NN	O	O
-	NN	O	O
18	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
gene	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
other	NN	O	O
than	NN	O	O
BRCA1	NN	O	O
which	NN	O	O
predisposes	NN	O	O
to	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
in	NN	O	O
women	NN	O	O
and	NN	O	O
which	NN	O	O
confers	NN	O	O
a	NN	O	O
higher	NN	O	O
risk	NN	O	O
of	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
additional	NN	O	O
pedigrees	NN	O	O
that	NN	O	O
include	NN	O	O
cases	NN	O	O
of	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
may	NN	O	O
therefore	NN	O	O
facilitate	NN	O	O
the	NN	O	O
mapping	NN	O	O
and	NN	O	O
isolation	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
EWS	NN	O	O
gene	NN	O	O
,	NN	O	O
involved	NN	O	O
in	NN	O	O
Ewing	NN	O	B-Disease
family	NN	O	I-Disease
of	NN	O	I-Disease
tumors	NN	O	I-Disease
,	NN	O	O
malignant	NN	O	B-Disease
melanoma	NN	O	I-Disease
of	NN	O	I-Disease
soft	NN	O	I-Disease
parts	NN	O	I-Disease
and	NN	O	O
desmoplastic	NN	O	B-Disease
small	NN	O	I-Disease
round	NN	O	I-Disease
cell	NN	O	I-Disease
tumors	NN	O	I-Disease
,	NN	O	O
codes	NN	O	O
for	NN	O	O
an	NN	O	O
RNA	NN	O	O
binding	NN	O	O
protein	NN	O	O
with	NN	O	O
novel	NN	O	O
regulatory	NN	O	O
domains	NN	O	O
.	NN	O	O

The	NN	O	O
EWS	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
maps	NN	O	O
to	NN	O	O
band	NN	O	O
q12	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
22	NN	O	O
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
human	NN	O	O
solid	NN	O	B-Disease
tumors	NN	O	I-Disease
including	NN	O	O
Ewing	NN	O	B-Disease
sarcoma	NN	O	I-Disease
,	NN	O	O
related	NN	O	O
primitive	NN	O	O
neuroectodermal	NN	O	B-Disease
tumors	NN	O	I-Disease
,	NN	O	O
malignant	NN	O	B-Disease
melanoma	NN	O	I-Disease
of	NN	O	I-Disease
soft	NN	O	I-Disease
parts	NN	O	I-Disease
and	NN	O	O
desmoplastic	NN	O	B-Disease
small	NN	O	I-Disease
round	NN	O	I-Disease
cell	NN	O	I-Disease
tumors	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
these	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
EWS	NN	O	O
is	NN	O	O
fused	NN	O	O
to	NN	O	O
genes	NN	O	O
encoding	NN	O	O
transcriptional	NN	O	O
activators	NN	O	O
/	NN	O	O
repressors	NN	O	O
,	NN	O	O
like	NN	O	O
Fli	NN	O	O
-	NN	O	O
1	NN	O	O
or	NN	O	O
erg	NN	O	O
or	NN	O	O
ATF	NN	O	O
1	NN	O	O
or	NN	O	O
wt1	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
EWS	NN	O	O
protein	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
EWS	NN	O	O
cDNA	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
this	NN	O	O
cDNA	NN	O	O
revealed	NN	O	O
differential	NN	O	O
splicing	NN	O	O
involving	NN	O	O
two	NN	O	O
exons	NN	O	O
encoding	NN	O	O
72	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

Both	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
transcripts	NN	O	O
,	NN	O	O
EWS	NN	O	O
and	NN	O	O
EWS	NN	O	O
-	NN	O	O
b	NN	O	O
,	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cells	NN	O	O
.	NN	O	O

Because	NN	O	O
EWS	NN	O	O
proteins	NN	O	O
contain	NN	O	O
putative	NN	O	O
conserved	NN	O	O
RNA	NN	O	O
binding	NN	O	O
motifs	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
RNA	NN	O	O
binding	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
EWS	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
EWS	NN	O	O
-	NN	O	O
b	NN	O	O
protein	NN	O	O
binds	NN	O	O
to	NN	O	O
RNA	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
,	NN	O	O
specifically	NN	O	O
,	NN	O	O
to	NN	O	O
poly	NN	O	O
G	NN	O	O
and	NN	O	O
poly	NN	O	O
U	NN	O	O
.	NN	O	O

The	NN	O	O
RNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
carboxy	NN	O	O
terminal	NN	O	O
86	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
which	NN	O	O
constitute	NN	O	O
RGG	NN	O	O
box	NN	O	O
.	NN	O	O

Thus	NN	O	O
the	NN	O	O
amino	NN	O	O
terminal	NN	O	O
domain	NN	O	O
of	NN	O	O
EWS	NN	O	O
(	NN	O	O
NTD	NN	O	O
-	NN	O	O
EWS	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
specificity	NN	O	O
of	NN	O	O
RNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
EWS	NN	O	O
.	NN	O	O

An	NN	O	O
EWS	NN	O	O
-	NN	O	O
erg	NN	O	O
chimeric	NN	O	O
protein	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
Ewings	NN	O	B-Disease
sarcoma	NN	O	I-Disease
cells	NN	O	O
,	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
EWS	NN	O	O
-	NN	O	O
erg	NN	O	O
chimeric	NN	O	O
protein	NN	O	O
revealed	NN	O	O
that	NN	O	O
NTD	NN	O	O
-	NN	O	O
EWS	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
regulatory	NN	O	O
domain	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
properties	NN	O	O
of	NN	O	O
EWS	NN	O	O
-	NN	O	O
erg	NN	O	O
chimeric	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
:	NN	O	O
genomic	NN	O	O
organization	NN	O	O
and	NN	O	O
localization	NN	O	O
of	NN	O	O
human	NN	O	O
ASPA	NN	O	O
to	NN	O	O
17p13	NN	O	O
-	NN	O	O
ter	NN	O	O
and	NN	O	O
conservation	NN	O	O
of	NN	O	O
the	NN	O	O
ASPA	NN	O	O
gene	NN	O	O
during	NN	O	O
evolution	NN	O	O
.	NN	O	O

Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
or	NN	O	O
spongy	NN	O	B-Disease
degeneration	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
brain	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
a	NN	O	O
severe	NN	O	O
leukodystrophy	NN	O	B-Disease
caused	NN	O	O
by	NN	O	O
the	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
aspartoacylase	NN	O	I-Disease
(	NN	O	O
ASPA	NN	O	O
)	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
human	NN	O	O
ASPA	NN	O	O
coding	NN	O	O
sequence	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
human	NN	O	O
ASPA	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
found	NN	O	O
to	NN	O	O
span	NN	O	O
29	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
genome	NN	O	O
.	NN	O	O

Human	NN	O	O
aspartoacylase	NN	O	O
is	NN	O	O
coded	NN	O	O
by	NN	O	O
six	NN	O	O
exons	NN	O	O
intervened	NN	O	O
by	NN	O	O
five	NN	O	O
introns	NN	O	O
.	NN	O	O

The	NN	O	O
exons	NN	O	O
vary	NN	O	O
from	NN	O	O
94	NN	O	O
(	NN	O	O
exon	NN	O	O
III	NN	O	O
)	NN	O	O
to	NN	O	O
514	NN	O	O
(	NN	O	O
exon	NN	O	O
VI	NN	O	O
)	NN	O	O
bases	NN	O	O
.	NN	O	O

The	NN	O	O
exon	NN	O	O
/	NN	O	O
intron	NN	O	O
splice	NN	O	O
junction	NN	O	O
sites	NN	O	O
follow	NN	O	O
the	NN	O	O
gt	NN	O	O
/	NN	O	O
ag	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
rule	NN	O	O
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
human	NN	O	O
/	NN	O	O
mouse	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrid	NN	O	O
cell	NN	O	O
lines	NN	O	O
localized	NN	O	O
ASPA	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
17	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
ASPA	NN	O	O
locus	NN	O	O
was	NN	O	O
further	NN	O	O
mapped	NN	O	O
in	NN	O	O
the	NN	O	O
17p13	NN	O	O
-	NN	O	O
ter	NN	O	O
region	NN	O	O
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

The	NN	O	O
bovine	NN	O	O
aspa	NN	O	O
gene	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
cloned	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
exon	NN	O	O
/	NN	O	O
intron	NN	O	O
organization	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
500	NN	O	O
-	NN	O	O
base	NN	O	O
sequence	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
initiator	NN	O	O
ATG	NN	O	O
codon	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
and	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
bovine	NN	O	O
gene	NN	O	O
are	NN	O	O
77	NN	O	O
%	NN	O	O
identical	NN	O	O
.	NN	O	O

Human	NN	O	O
ASPA	NN	O	O
coding	NN	O	O
sequences	NN	O	O
cross	NN	O	O
-	NN	O	O
hybridize	NN	O	O
with	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
from	NN	O	O
yeast	NN	O	O
,	NN	O	O
chicken	NN	O	O
,	NN	O	O
rabbit	NN	O	O
,	NN	O	O
cow	NN	O	O
,	NN	O	O
dog	NN	O	O
,	NN	O	O
mouse	NN	O	O
,	NN	O	O
rat	NN	O	O
,	NN	O	O
and	NN	O	O
monkey	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
cross	NN	O	O
-	NN	O	O
species	NN	O	O
hybridization	NN	O	O
of	NN	O	O
coding	NN	O	O
sequences	NN	O	O
suggests	NN	O	O
that	NN	O	O
aspartoacylase	NN	O	O
has	NN	O	O
been	NN	O	O
conserved	NN	O	O
during	NN	O	O
evolution	NN	O	O
.	NN	O	O

It	NN	O	O
should	NN	O	O
now	NN	O	O
be	NN	O	O
possible	NN	O	O
to	NN	O	O
identify	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
noncoding	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
Canavan	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
ASPA	NN	O	O
.	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
:	NN	O	O
size	NN	O	O
-	NN	O	O
and	NN	O	O
sex	NN	O	O
-	NN	O	O
dependent	NN	O	O
dynamics	NN	O	O
of	NN	O	O
CTG	NN	O	O
meiotic	NN	O	O
instability	NN	O	O
,	NN	O	O
and	NN	O	O
somatic	NN	O	O
mosaicism	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
progressive	NN	O	O
neuromuscular	NN	O	B-Disease
disorder	NN	O	I-Disease
which	NN	O	O
results	NN	O	O
from	NN	O	O
elongations	NN	O	O
of	NN	O	O
an	NN	O	O
unstable	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

A	NN	O	O
correlation	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
between	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
repeat	NN	O	O
number	NN	O	O
of	NN	O	O
this	NN	O	O
sequence	NN	O	O
and	NN	O	O
the	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
clinical	NN	O	O
status	NN	O	O
of	NN	O	O
patients	NN	O	O
cannot	NN	O	O
be	NN	O	O
unambiguously	NN	O	O
ascertained	NN	O	O
solely	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
CTG	NN	O	O
repeats	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
exclusive	NN	O	O
maternal	NN	O	O
inheritance	NN	O	O
of	NN	O	O
the	NN	O	O
congenital	NN	O	O
form	NN	O	O
remains	NN	O	O
unexplained	NN	O	O
.	NN	O	O

Our	NN	O	O
observation	NN	O	O
of	NN	O	O
differently	NN	O	O
sized	NN	O	O
repeats	NN	O	O
in	NN	O	O
various	NN	O	O
DM	NN	O	B-Disease
tissues	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
individual	NN	O	O
may	NN	O	O
explain	NN	O	O
why	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
observed	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
does	NN	O	O
not	NN	O	O
necessarily	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
severity	NN	O	O
and	NN	O	O
nature	NN	O	O
of	NN	O	O
symptoms	NN	O	O
.	NN	O	O

Through	NN	O	O
a	NN	O	O
molecular	NN	O	O
and	NN	O	O
genetic	NN	O	O
study	NN	O	O
of	NN	O	O
142	NN	O	O
families	NN	O	O
including	NN	O	O
418	NN	O	O
DM	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
dynamics	NN	O	O
of	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
meiotic	NN	O	O
instability	NN	O	O
.	NN	O	O

A	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
and	NN	O	O
the	NN	O	O
intergenerational	NN	O	O
enlargement	NN	O	O
was	NN	O	O
observed	NN	O	O
similarly	NN	O	O
through	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
meioses	NN	O	O
for	NN	O	O
<	NN	O	O
or	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
-	NN	O	O
kb	NN	O	O
CTG	NN	O	O
sequences	NN	O	O
.	NN	O	O

Beyond	NN	O	O
0	NN	O	O
.	NN	O	O

5	NN	O	O
kb	NN	O	O
,	NN	O	O
the	NN	O	O
intergenerational	NN	O	O
variation	NN	O	O
was	NN	O	O
more	NN	O	O
important	NN	O	O
through	NN	O	O
female	NN	O	O
meioses	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
tendency	NN	O	O
to	NN	O	O
compression	NN	O	O
was	NN	O	O
observed	NN	O	O
almost	NN	O	O
exclusively	NN	O	O
in	NN	O	O
male	NN	O	O
meioses	NN	O	O
,	NN	O	O
for	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
1	NN	O	O
.	NN	O	O

5	NN	O	O
-	NN	O	O
kb	NN	O	O
fragments	NN	O	O
.	NN	O	O

This	NN	O	O
implies	NN	O	O
a	NN	O	O
size	NN	O	O
-	NN	O	O
and	NN	O	O
sex	NN	O	O
-	NN	O	O
dependent	NN	O	O
meiotic	NN	O	O
instability	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
segregation	NN	O	O
analysis	NN	O	O
supports	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
a	NN	O	O
maternal	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
familial	NN	O	O
predisposition	NN	O	O
for	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
the	NN	O	O
congenital	NN	O	O
form	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
this	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
a	NN	O	O
significant	NN	O	O
excess	NN	O	O
of	NN	O	O
transmitting	NN	O	O
grandfathers	NN	O	O
partially	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
increased	NN	O	O
fertility	NN	O	O
in	NN	O	O
affected	NN	O	O
males	NN	O	O

Illegitimate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
gene	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
for	NN	O	O
identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O

Taking	NN	O	O
advantage	NN	O	O
of	NN	O	O
the	NN	O	O
illegitimate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase	NN	O	O
(	NN	O	O
PAH	NN	O	O
)	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
been	NN	O	O
able	NN	O	O
to	NN	O	O
analyse	NN	O	O
the	NN	O	O
PAH	NN	O	O
cDNA	NN	O	O
sequence	NN	O	O
of	NN	O	O
hyperphenylalaninemic	NN	O	B-Disease
children	NN	O	O
in	NN	O	O
circulating	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
approach	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
identified	NN	O	O
3	NN	O	O
novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
cDNA	NN	O	O
from	NN	O	O
liver	NN	O	O
and	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
two	NN	O	O
patients	NN	O	O
.	NN	O	O

One	NN	O	O
mutation	NN	O	O
,	NN	O	O
detected	NN	O	O
by	NN	O	O
the	NN	O	O
abnormal	NN	O	O
pattern	NN	O	O
of	NN	O	O
migration	NN	O	O
of	NN	O	O
an	NN	O	O
amplified	NN	O	O
fragment	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
C	NN	O	O
to	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
the	NN	O	O
splice	NN	O	O
acceptor	NN	O	O
site	NN	O	O
of	NN	O	O
intron	NN	O	O
10	NN	O	O
,	NN	O	O
which	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
skipping	NN	O	O
of	NN	O	O
exon	NN	O	O
11	NN	O	O
with	NN	O	O
the	NN	O	O
premature	NN	O	O
termination	NN	O	O
of	NN	O	O
RNA	NN	O	O
translation	NN	O	O
downstream	NN	O	O
from	NN	O	O
exon	NN	O	O
12	NN	O	O
(	NN	O	O
-	NN	O	O
3	NN	O	O
IVS10	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
two	NN	O	O
mutations	NN	O	O
are	NN	O	O
missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
exons	NN	O	O
10	NN	O	O
and	NN	O	O
11	NN	O	O
(	NN	O	O
respectively	NN	O	O
,	NN	O	O
L333F	NN	O	O
and	NN	O	O
E390G	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
supports	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
circulating	NN	O	O
lymphocytes	NN	O	O
give	NN	O	O
easy	NN	O	O
access	NN	O	O
to	NN	O	O
PAH	NN	O	O
gene	NN	O	O
transcripts	NN	O	O
whose	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
reported	NN	O	O
in	NN	O	O
liver	NN	O	O
and	NN	O	O
therefore	NN	O	O
represent	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
studies	NN	O	O
in	NN	O	O
phenylketonuria	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

High	NN	O	O
residual	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
infantile	NN	O	I-Disease
metachromatic	NN	O	I-Disease
leukodystrophy	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
patient	NN	O	O
suffering	NN	O	O
from	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
infantile	NN	O	I-Disease
metachromatic	NN	O	I-Disease
leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
who	NN	O	O
has	NN	O	O
a	NN	O	O
residual	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
activity	NN	O	O
of	NN	O	O
about	NN	O	O
10	NN	O	O
%	NN	O	O
.	NN	O	O

Fibroblasts	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
show	NN	O	O
significant	NN	O	O
sulfatide	NN	O	O
degradation	NN	O	O
activity	NN	O	O
exceeding	NN	O	O
that	NN	O	O
of	NN	O	O
adult	NN	O	B-Disease
MLD	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
ARSA	NN	O	O
gene	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
revealed	NN	O	O
heterozygosity	NN	O	O
for	NN	O	O
two	NN	O	O
new	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
in	NN	O	O
one	NN	O	O
allele	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
C	NN	O	O
447	NN	O	O
in	NN	O	O
exon	NN	O	O
2	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
frameshift	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
position	NN	O	O
105	NN	O	O
;	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
allele	NN	O	O
,	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
in	NN	O	O
exon	NN	O	O
5	NN	O	O
causes	NN	O	O
a	NN	O	O
Gly309	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Ser	NN	O	O
substitution	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
Ser309	NN	O	O
-	NN	O	O
ARSA	NN	O	O
resulted	NN	O	O
in	NN	O	O
only	NN	O	O
13	NN	O	O
%	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
of	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
normal	NN	O	O
ARSA	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
ARSA	NN	O	O
is	NN	O	O
correctly	NN	O	O
targeted	NN	O	O
to	NN	O	O
the	NN	O	O
lysosomes	NN	O	O
but	NN	O	O
is	NN	O	O
unstable	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
previous	NN	O	O
results	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
infantile	NN	O	I-Disease
type	NN	O	I-Disease
of	NN	O	I-Disease
MLD	NN	O	I-Disease
is	NN	O	O
always	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
complete	NN	O	O
absence	NN	O	O
of	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
ARSA	NN	O	O
protein	NN	O	O
may	NN	O	O
be	NN	O	O
influenced	NN	O	O
by	NN	O	O
particular	NN	O	O
features	NN	O	O
of	NN	O	O
oligodendrocytes	NN	O	O
,	NN	O	O
such	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
mutant	NN	O	O
enzyme	NN	O	O
is	NN	O	O
lower	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
than	NN	O	O
in	NN	O	O
others	NN	O	O
.	NN	O	O
.	NN	O	O

An	NN	O	O
arylsulfatase	NN	O	O
A	NN	O	O
(	NN	O	O
ARSA	NN	O	O
)	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
T274M	NN	O	O
)	NN	O	O
causing	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
infantile	NN	O	I-Disease
metachromatic	NN	O	I-Disease
leukodystrophy	NN	O	I-Disease
.	NN	O	O

Metachromatic	NN	O	B-Disease
leukodystrophy	NN	O	I-Disease
(	NN	O	O
MLD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
lysosomal	NN	O	I-Disease
storage	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
arylsulfatase	NN	O	I-Disease
A	NN	O	I-Disease
(	NN	O	O
ARSA	NN	O	O
;	NN	O	O
EC	NN	O	O
3	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O
6	NN	O	O
.	NN	O	O
8	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
8	NN	O	O
ARSA	NN	O	O
exons	NN	O	O
and	NN	O	O
adjacent	NN	O	O
intron	NN	O	O
boundaries	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
late	NN	O	B-Disease
-	NN	O	I-Disease
infantile	NN	O	I-Disease
metachromatic	NN	O	I-Disease
leukodystrophy	NN	O	I-Disease
were	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
amplified	NN	O	O
in	NN	O	O
seven	NN	O	O
discrete	NN	O	O
reactions	NN	O	O
.	NN	O	O

Amplified	NN	O	O
ARSA	NN	O	O
exons	NN	O	O
were	NN	O	O
analysed	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sequence	NN	O	O
alterations	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
PCR	NN	O	O
products	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
in	NN	O	O
exon	NN	O	O
IV	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
threonine	NN	O	O
residue	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
274	NN	O	O
with	NN	O	O
a	NN	O	O
methionine	NN	O	O
(	NN	O	O
T274M	NN	O	O
)	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
a	NN	O	O
further	NN	O	O
29	NN	O	O
MLD	NN	O	B-Disease
patients	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
five	NN	O	O
additional	NN	O	O
homozygotes	NN	O	O
for	NN	O	O
T274M	NN	O	O
.	NN	O	O

All	NN	O	O
6	NN	O	O
T274M	NN	O	O
homozygotes	NN	O	O
(	NN	O	O
representing	NN	O	O
four	NN	O	O
families	NN	O	O
)	NN	O	O
were	NN	O	O
of	NN	O	O
Lebanese	NN	O	O
descent	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
were	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
consanguineous	NN	O	O
marriages	NN	O	O
.	NN	O	O

The	NN	O	O
altered	NN	O	O
amino	NN	O	O
acid	NN	O	O
is	NN	O	O
rigidly	NN	O	O
conserved	NN	O	O
among	NN	O	O
10	NN	O	O
sulfatases	NN	O	O
from	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
to	NN	O	O
humans	NN	O	O
;	NN	O	O
therefore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
most	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
resultant	NN	O	O
mutant	NN	O	O
protein	NN	O	O
will	NN	O	O
have	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
very	NN	O	O
low	NN	O	O
ARSA	NN	O	O
activity	NN	O	O
measured	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
and	NN	O	O
their	NN	O	O
uniformly	NN	O	O
severe	NN	O	O
clinical	NN	O	O
presentation	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
hereditary	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
14	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
analysed	NN	O	O
exon	NN	O	O
-	NN	O	O
by	NN	O	O
-	NN	O	O
exon	NN	O	O
using	NN	O	O
SSCP	NN	O	O
in	NN	O	O
6	NN	O	O
aniridia	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

In	NN	O	O
each	NN	O	O
family	NN	O	O
band	NN	O	O
shifts	NN	O	O
were	NN	O	O
observed	NN	O	O
on	NN	O	O
the	NN	O	O
SSCP	NN	O	O
gels	NN	O	O
for	NN	O	O
only	NN	O	O
one	NN	O	O
exon	NN	O	O
and	NN	O	O
direct	NN	O	O
PCR	NN	O	O
-	NN	O	O
sequencing	NN	O	O
revealed	NN	O	O
mutations	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
.	NN	O	O

Two	NN	O	O
mutations	NN	O	O
involved	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transitions	NN	O	O
in	NN	O	O
CGAarg	NN	O	O
codons	NN	O	O
in	NN	O	O
exons	NN	O	O
9	NN	O	O
and	NN	O	O
11	NN	O	O
.	NN	O	O

Another	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
converted	NN	O	O
a	NN	O	O
CAG	NN	O	O
-	NN	O	O
glutamine	NN	O	O
to	NN	O	O
a	NN	O	O
TAG	NN	O	O
-	NN	O	O
stop	NN	O	O
in	NN	O	O
exon	NN	O	O
7	NN	O	O
.	NN	O	O

Small	NN	O	O
insertions	NN	O	O
created	NN	O	O
frameshifts	NN	O	O
which	NN	O	O
produced	NN	O	O
downstream	NN	O	O
stop	NN	O	O
codons	NN	O	O
in	NN	O	O
another	NN	O	O
two	NN	O	O
patients	NN	O	O
and	NN	O	O
an	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
mutation	NN	O	O
disrupted	NN	O	O
the	NN	O	O
splice	NN	O	O
donor	NN	O	O
site	NN	O	O
of	NN	O	O
exon	NN	O	O
5	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
family	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
complete	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
is	NN	O	O
predicted	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
other	NN	O	O
affected	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
showed	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
,	NN	O	O
all	NN	O	O
affected	NN	O	O
individuals	NN	O	O
carried	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
-	NN	O	O
specific	NN	O	O
mutation	NN	O	O
.	NN	O	O

One	NN	O	O
polymorphism	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
exon	NN	O	O
7	NN	O	O
.	NN	O	O

This	NN	O	O
data	NN	O	O
strongly	NN	O	O
supports	NN	O	O
the	NN	O	O
candidature	NN	O	O
of	NN	O	O
PAX6	NN	O	O
as	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
hereditary	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Three	NN	O	O
novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
five	NN	O	O
unrelated	NN	O	O
subjects	NN	O	O
with	NN	O	O
hereditary	NN	O	O
protein	NN	O	B-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
type	NN	O	I-Disease
I	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
panel	NN	O	O
of	NN	O	O
eight	NN	O	O
unrelated	NN	O	O
subjects	NN	O	O
with	NN	O	O
inherited	NN	O	O
type	NN	O	B-Disease
I	NN	O	I-Disease
protein	NN	O	I-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
was	NN	O	O
screened	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
PROS1	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
five	NN	O	O
subjects	NN	O	O
an	NN	O	O
abnormality	NN	O	O
was	NN	O	O
found	NN	O	O
but	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
three	NN	O	O
subjects	NN	O	O
.	NN	O	O

Two	NN	O	O
subjects	NN	O	O
shared	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
at	NN	O	O
position	NN	O	O
+	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
of	NN	O	O
intron	NN	O	O
10	NN	O	O
.	NN	O	O

Also	NN	O	O
in	NN	O	O
two	NN	O	O
subjects	NN	O	O
an	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transversion	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
stopcodon	NN	O	O
of	NN	O	O
the	NN	O	O
PROS1	NN	O	O
gene	NN	O	O
;	NN	O	O
this	NN	O	O
transversion	NN	O	O
predicts	NN	O	O
a	NN	O	O
protein	NN	O	O
S	NN	O	O
molecule	NN	O	O
that	NN	O	O
is	NN	O	O
extended	NN	O	O
by	NN	O	O
14	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

The	NN	O	O
fifth	NN	O	O
subject	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
possess	NN	O	O
two	NN	O	O
sequence	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

One	NN	O	O
allele	NN	O	O
carried	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
transition	NN	O	O
near	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
junction	NN	O	O
of	NN	O	O
intron	NN	O	O
2	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
abnormality	NN	O	O
is	NN	O	O
probably	NN	O	O
neutral	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
was	NN	O	O
inherited	NN	O	O
from	NN	O	O
the	NN	O	O
parent	NN	O	O
with	NN	O	O
normal	NN	O	O
protein	NN	O	O
S	NN	O	O
antigen	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
other	NN	O	O
allele	NN	O	O
a	NN	O	O
single	NN	O	O
T	NN	O	O
insertion	NN	O	O
in	NN	O	O
codon	NN	O	O
-	NN	O	O
25	NN	O	O
was	NN	O	O
found	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
platelet	NN	O	O
RNA	NN	O	O
showed	NN	O	O
that	NN	O	O
only	NN	O	O
the	NN	O	O
mRNA	NN	O	O
with	NN	O	O
the	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
stopcodon	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
amounts	NN	O	O
comparable	NN	O	O
to	NN	O	O
wildtype	NN	O	O
RNA	NN	O	O
.	NN	O	O

mRNA	NN	O	O
from	NN	O	O
the	NN	O	O
alleles	NN	O	O
with	NN	O	O
the	NN	O	O
other	NN	O	O
two	NN	O	O
mutations	NN	O	O
was	NN	O	O
either	NN	O	O
undetectable	NN	O	O
or	NN	O	O
present	NN	O	O
in	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
amounts	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
indicates	NN	O	O
that	NN	O	O
a	NN	O	O
mRNA	NN	O	O
based	NN	O	O
approach	NN	O	O
is	NN	O	O
not	NN	O	O
feasible	NN	O	O
for	NN	O	O
the	NN	O	O
genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
protein	NN	O	B-Disease
S	NN	O	I-Disease
deficiency	NN	O	I-Disease
type	NN	O	I-Disease
I	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Characteristics	NN	O	O
of	NN	O	O
intergenerational	NN	O	O
contractions	NN	O	O
of	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
a	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
generally	NN	O	O
increases	NN	O	O
in	NN	O	O
successive	NN	O	O
generations	NN	O	O
with	NN	O	O
clinical	NN	O	O
evidence	NN	O	O
of	NN	O	O
anticipation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
have	NN	O	O
also	NN	O	O
been	NN	O	O
cases	NN	O	O
with	NN	O	O
an	NN	O	O
intergenerational	NN	O	O
contraction	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
1	NN	O	O
,	NN	O	O
489	NN	O	O
DM	NN	O	B-Disease
parent	NN	O	O
-	NN	O	O
offspring	NN	O	O
pairs	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
95	NN	O	O
(	NN	O	O
6	NN	O	O
.	NN	O	O
4	NN	O	O
%	NN	O	O
)	NN	O	O
showed	NN	O	O
such	NN	O	O
contractions	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
(	NN	O	O
PBL	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
56	NN	O	O
of	NN	O	O
the	NN	O	O
95	NN	O	O
pairs	NN	O	O
,	NN	O	O
clinical	NN	O	O
data	NN	O	O
allowed	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
their	NN	O	O
anticipation	NN	O	O
status	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
surprising	NN	O	O
that	NN	O	O
anticipation	NN	O	O
occurred	NN	O	O
in	NN	O	O
27	NN	O	O
(	NN	O	O
48	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
these	NN	O	O
56	NN	O	O
pairs	NN	O	O
,	NN	O	O
while	NN	O	O
none	NN	O	O
clearly	NN	O	O
showed	NN	O	O
a	NN	O	O
later	NN	O	O
onset	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
symptomatic	NN	O	O
offspring	NN	O	O
.	NN	O	O

The	NN	O	O
contraction	NN	O	O
occurred	NN	O	O
in	NN	O	O
76	NN	O	O
(	NN	O	O
10	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
753	NN	O	O
paternal	NN	O	O
transmissions	NN	O	O
and	NN	O	O
in	NN	O	O
19	NN	O	O
(	NN	O	O
3	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
736	NN	O	O
maternal	NN	O	O
transmissions	NN	O	O
.	NN	O	O

Anticipation	NN	O	O
was	NN	O	O
observed	NN	O	O
more	NN	O	O
frequently	NN	O	O
in	NN	O	O
maternal	NN	O	O
(	NN	O	O
85	NN	O	O
%	NN	O	O
)	NN	O	O
than	NN	O	O
in	NN	O	O
paternal	NN	O	O
(	NN	O	O
37	NN	O	O
%	NN	O	O
)	NN	O	O
transmissions	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
parental	NN	O	O
repeat	NN	O	O
size	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
intergenerational	NN	O	O
contraction	NN	O	O
(	NN	O	O
r2	NN	O	O
=	NN	O	O
.	NN	O	O
50	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
<	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
slope	NN	O	O
of	NN	O	O
linear	NN	O	O
regression	NN	O	O
was	NN	O	O
steeper	NN	O	O
in	NN	O	O
paternal	NN	O	O
(	NN	O	O
-	NN	O	O
.	NN	O	O
62	NN	O	O
)	NN	O	O
than	NN	O	O
in	NN	O	O
maternal	NN	O	O
(	NN	O	O
-	NN	O	O
.	NN	O	O
30	NN	O	O
)	NN	O	O
transmissions	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
<	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
.	NN	O	O

Sixteen	NN	O	O
DM	NN	O	B-Disease
parents	NN	O	O
had	NN	O	O
multiple	NN	O	O
DM	NN	O	B-Disease
offspring	NN	O	O
with	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
contractions	NN	O	O
.	NN	O	O

This	NN	O	O
frequency	NN	O	O
was	NN	O	O
higher	NN	O	O
than	NN	O	O
the	NN	O	O
frequency	NN	O	O
expected	NN	O	O
from	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
contractions	NN	O	O
(	NN	O	O
6	NN	O	O
.	NN	O	O
4	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
sib	NN	O	O
population	NN	O	O
(	NN	O	O
1	NN	O	O
.	NN	O	O
54	NN	O	O
DM	NN	O	B-Disease
offspring	NN	O	O
per	NN	O	O
DM	NN	O	B-Disease
parent	NN	O	O
,	NN	O	O
in	NN	O	O
968	NN	O	O
DM	NN	O	B-Disease
parents	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
intergenerational	NN	O	O
contractions	NN	O	O
of	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
in	NN	O	O
leukocyte	NN	O	O
DNA	NN	O	O
frequently	NN	O	O
accompanies	NN	O	O
apparent	NN	O	O
anticipation	NN	O	O
,	NN	O	O
especially	NN	O	O
when	NN	O	O
DM	NN	O	B-Disease
is	NN	O	O
maternally	NN	O	O
transmitted	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
paternal	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
repeat	NN	O	O
contraction	NN	O	O
in	NN	O	O
a	NN	O	O
sibling	NN	O	O
increase	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
the	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
contraction	NN	O	O

Gonosomal	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
patients	NN	O	O
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
mitotic	NN	O	O
events	NN	O	O
in	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
variation	NN	O	O
and	NN	O	O
selection	NN	O	O
against	NN	O	O
extreme	NN	O	O
expansion	NN	O	O
in	NN	O	O
sperm	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
abnormal	NN	O	O
expansion	NN	O	O
of	NN	O	O
a	NN	O	O
polymorphic	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
protein	NN	O	O
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
the	NN	O	O
(	NN	O	O
CTG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
lengths	NN	O	O
in	NN	O	O
a	NN	O	O
broad	NN	O	O
range	NN	O	O
of	NN	O	O
tissue	NN	O	O
DNAs	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
mild	NN	O	O
,	NN	O	O
classical	NN	O	O
,	NN	O	O
or	NN	O	O
congenital	NN	O	O
manifestation	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
the	NN	O	O
repeat	NN	O	O
length	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
somatic	NN	O	O
tissues	NN	O	O
from	NN	O	O
single	NN	O	O
DM	NN	O	B-Disease
individuals	NN	O	O
and	NN	O	O
twins	NN	O	O
.	NN	O	O

Repeats	NN	O	O
appeared	NN	O	O
to	NN	O	O
expand	NN	O	O
to	NN	O	O
a	NN	O	O
similar	NN	O	O
extent	NN	O	O
in	NN	O	O
tissues	NN	O	O
originating	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
embryonal	NN	O	O
origin	NN	O	O
.	NN	O	O

In	NN	O	O
most	NN	O	O
male	NN	O	O
patients	NN	O	O
carrying	NN	O	O
intermediate	NN	O	O
-	NN	O	O
or	NN	O	O
small	NN	O	O
-	NN	O	O
sized	NN	O	O
expansions	NN	O	O
in	NN	O	O
blood	NN	O	O
,	NN	O	O
the	NN	O	O
repeat	NN	O	O
lengths	NN	O	O
covered	NN	O	O
a	NN	O	O
markedly	NN	O	O
wider	NN	O	O
range	NN	O	O
in	NN	O	O
sperm	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
male	NN	O	O
patients	NN	O	O
with	NN	O	O
large	NN	O	O
allele	NN	O	O
expansions	NN	O	O
in	NN	O	O
blood	NN	O	O
(	NN	O	O
>	NN	O	O
700	NN	O	O
CTGs	NN	O	O
)	NN	O	O
had	NN	O	O
similar	NN	O	O
or	NN	O	O
smaller	NN	O	O
repeats	NN	O	O
in	NN	O	O
sperm	NN	O	O
,	NN	O	O
when	NN	O	O
detectable	NN	O	O
.	NN	O	O

Sperm	NN	O	O
alleles	NN	O	O
with	NN	O	O
>	NN	O	O
1	NN	O	O
,	NN	O	O
000	NN	O	O
CTGs	NN	O	O
were	NN	O	O
not	NN	O	O
seen	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
DM	NN	O	B-Disease
patients	NN	O	O
can	NN	O	O
be	NN	O	O
considered	NN	O	O
gonosomal	NN	O	O
mosaics	NN	O	O
,	NN	O	O
i	NN	O	O
.	NN	O	O

e	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
combined	NN	O	O
somatic	NN	O	O
and	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
tissue	NN	O	O
mosaics	NN	O	O
.	NN	O	O

Most	NN	O	O
remarkably	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
multiple	NN	O	O
cases	NN	O	O
where	NN	O	O
the	NN	O	O
length	NN	O	O
distributions	NN	O	O
of	NN	O	O
intermediate	NN	O	O
-	NN	O	O
or	NN	O	O
small	NN	O	O
-	NN	O	O
sized	NN	O	O
alleles	NN	O	O
in	NN	O	O
fathers	NN	O	O
sperm	NN	O	O
were	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
in	NN	O	O
their	NN	O	O
offsprings	NN	O	O
blood	NN	O	O
.	NN	O	O

Our	NN	O	O
combined	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
intergenerational	NN	O	O
length	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
unstable	NN	O	O
CTG	NN	O	O
repeat	NN	O	O
are	NN	O	O
most	NN	O	O
likely	NN	O	O
to	NN	O	O
occur	NN	O	O
during	NN	O	O
early	NN	O	O
embryonic	NN	O	O
mitotic	NN	O	O
divisions	NN	O	O
in	NN	O	O
both	NN	O	O
somatic	NN	O	O
and	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
tissue	NN	O	O
formation	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
initial	NN	O	O
CTG	NN	O	O
length	NN	O	O
,	NN	O	O
the	NN	O	O
overall	NN	O	O
number	NN	O	O
of	NN	O	O
cell	NN	O	O
divisions	NN	O	O
involved	NN	O	O
in	NN	O	O
tissue	NN	O	O
formation	NN	O	O
,	NN	O	O
and	NN	O	O
perhaps	NN	O	O
a	NN	O	O
specific	NN	O	O
selection	NN	O	O
process	NN	O	O
in	NN	O	O
spermatogenesis	NN	O	O
may	NN	O	O
influence	NN	O	O
the	NN	O	O
dynamics	NN	O	O
of	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

A	NN	O	O
model	NN	O	O
explaining	NN	O	O
mitotic	NN	O	O
instability	NN	O	O
and	NN	O	O
sex	NN	O	O
-	NN	O	O
dependent	NN	O	O
segregation	NN	O	O
phenomena	NN	O	O
in	NN	O	O
DM	NN	O	B-Disease
manifestation	NN	O	O
is	NN	O	O
discussed	NN	O	O

Regionally	NN	O	O
clustered	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
and	NN	O	O
occur	NN	O	O
at	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
in	NN	O	O
new	NN	O	O
mutation	NN	O	O
cases	NN	O	O
of	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
.	NN	O	O

Germline	NN	O	O
mutation	NN	O	O
in	NN	O	O
APC	NN	O	O
at	NN	O	O
5q21	NN	O	O
-	NN	O	O
22	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
dominantly	NN	O	O
inherited	NN	O	O
syndrome	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
(	NN	O	O
APC	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Somatic	NN	O	O
mutation	NN	O	O
in	NN	O	O
this	NN	O	O
gene	NN	O	O
is	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
colorectal	NN	O	O
tumourigenesis	NN	O	O
.	NN	O	O

Both	NN	O	O
types	NN	O	O
of	NN	O	O
mutation	NN	O	O
are	NN	O	O
concentrated	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
half	NN	O	O
of	NN	O	O
exon	NN	O	O
15	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
single	NN	O	O
strand	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
and	NN	O	O
heteroduplex	NN	O	O
analysis	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
variants	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
45	NN	O	O
affected	NN	O	O
but	NN	O	O
unrelated	NN	O	O
individuals	NN	O	O
.	NN	O	O

Eighteen	NN	O	O
patients	NN	O	O
had	NN	O	O
no	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
;	NN	O	O
of	NN	O	O
these	NN	O	O
11	NN	O	O
were	NN	O	O
classified	NN	O	O
as	NN	O	O
having	NN	O	O
a	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
an	NN	O	O
early	NN	O	O
age	NN	O	O
at	NN	O	O
presentation	NN	O	O
or	NN	O	O
cancer	NN	O	B-Disease
development	NN	O	O
.	NN	O	O

This	NN	O	O
compared	NN	O	O
with	NN	O	O
6	NN	O	O
of	NN	O	O
27	NN	O	O
familial	NN	O	O
cases	NN	O	O
.	NN	O	O

A	NN	O	O
5	NN	O	O
bp	NN	O	O
deletion	NN	O	O
at	NN	O	O
codon	NN	O	O
1309	NN	O	O
reported	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
10	NN	O	O
-	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
unselected	NN	O	O
APC	NN	O	B-Disease
patients	NN	O	O
worldwide	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
5	NN	O	O
of	NN	O	O
the	NN	O	O
18	NN	O	O
new	NN	O	O
mutation	NN	O	O
cases	NN	O	O
and	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
27	NN	O	O
familial	NN	O	O
cases	NN	O	O
all	NN	O	O
nine	NN	O	O
were	NN	O	O
classed	NN	O	O
as	NN	O	O
severe	NN	O	O
.	NN	O	O

A	NN	O	O
further	NN	O	O
3	NN	O	O
new	NN	O	O
mutations	NN	O	O
and	NN	O	O
1	NN	O	O
familial	NN	O	O
mutation	NN	O	O
were	NN	O	O
located	NN	O	O
downstream	NN	O	O
from	NN	O	O
codon	NN	O	O
1309	NN	O	O
,	NN	O	O
these	NN	O	O
individuals	NN	O	O
similarly	NN	O	O
being	NN	O	O
classed	NN	O	O
as	NN	O	O
phenotypically	NN	O	O
severe	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
detected	NN	O	O
in	NN	O	O
affected	NN	O	O
individuals	NN	O	O
with	NN	O	O
an	NN	O	O
average	NN	O	O
phenotype	NN	O	O
were	NN	O	O
located	NN	O	O
prior	NN	O	O
to	NN	O	O
codon	NN	O	O
1309	NN	O	O
.	NN	O	O

The	NN	O	O
frequent	NN	O	O
association	NN	O	O
of	NN	O	O
a	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
with	NN	O	O
fresh	NN	O	O
mutation	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
apparent	NN	O	O
conflict	NN	O	O
of	NN	O	O
a	NN	O	O
high	NN	O	O
mutation	NN	O	O
rate	NN	O	O
(	NN	O	O
20	NN	O	O
-	NN	O	O
30	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
condition	NN	O	O
,	NN	O	O
which	NN	O	O
on	NN	O	O
average	NN	O	O
,	NN	O	O
is	NN	O	O
lethal	NN	O	O
at	NN	O	O
a	NN	O	O
post	NN	O	O
-	NN	O	O
reproductive	NN	O	O
age	NN	O	O
.	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
PAX6	NN	O	O
locus	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
heterogeneous	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
malformations	NN	O	I-Disease
including	NN	O	O
Peters	NN	O	B-Disease
'	NN	O	I-Disease
anomaly	NN	O	I-Disease
.	NN	O	O

Mutation	NN	O	O
or	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
underlies	NN	O	O
many	NN	O	O
cases	NN	O	O
of	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O

Three	NN	O	O
lines	NN	O	O
of	NN	O	O
evidence	NN	O	O
now	NN	O	O
converge	NN	O	O
to	NN	O	O
implicate	NN	O	O
PAX6	NN	O	O
more	NN	O	O
widely	NN	O	O
in	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
malformations	NN	O	I-Disease
including	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
.	NN	O	O

First	NN	O	O
,	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
is	NN	O	O
deleted	NN	O	O
for	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
PAX6	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
affected	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
with	NN	O	O
dominantly	NN	O	O
inherited	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
malformations	NN	O	I-Disease
,	NN	O	O
including	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
are	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
an	NN	O	O
R26G	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
PAX6	NN	O	O
paired	NN	O	O
box	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
a	NN	O	O
proportion	NN	O	O
of	NN	O	O
Sey	NN	O	O
/	NN	O	O
+	NN	O	O
Smalleye	NN	O	O
mice	NN	O	O
,	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
murine	NN	O	O
Pax	NN	O	O
-	NN	O	O
6	NN	O	O
,	NN	O	O
have	NN	O	O
an	NN	O	O
ocular	NN	O	O
phenotype	NN	O	O
resembling	NN	O	O
Peters	NN	O	B-Disease
anomaly	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
propose	NN	O	O
that	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
anterior	NN	O	B-Disease
segment	NN	O	I-Disease
anomalies	NN	O	I-Disease
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
PAX6	NN	O	O
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
without	NN	O	O
CAG	NN	O	O
expansion	NN	O	O
:	NN	O	O
phenocopies	NN	O	O
or	NN	O	O
errors	NN	O	O
in	NN	O	O
assignment	NN	O	O
?	NN	O	O

Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
expanded	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
within	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
on	NN	O	O
4p16	NN	O	O
.	NN	O	O

3	NN	O	O
(	NN	O	O
IT15	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
30	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
022	NN	O	O
affected	NN	O	O
persons	NN	O	O
(	NN	O	O
2	NN	O	O
.	NN	O	O
9	NN	O	O
%	NN	O	O
of	NN	O	O
our	NN	O	O
cohort	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
an	NN	O	O
expanded	NN	O	O
CAG	NN	O	O
in	NN	O	O
the	NN	O	O
disease	NN	O	O
range	NN	O	O
.	NN	O	O

The	NN	O	O
reasons	NN	O	O
for	NN	O	O
not	NN	O	O
observing	NN	O	O
expansion	NN	O	O
in	NN	O	O
affected	NN	O	O
individuals	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
determining	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
using	NN	O	O
repeat	NN	O	O
length	NN	O	O
both	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
affected	NN	O	O
patients	NN	O	O
and	NN	O	O
for	NN	O	O
predictive	NN	O	O
testing	NN	O	O
programs	NN	O	O
and	NN	O	O
may	NN	O	O
have	NN	O	O
biological	NN	O	O
relevance	NN	O	O
for	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
majority	NN	O	O
(	NN	O	O
18	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
individuals	NN	O	O
with	NN	O	O
normal	NN	O	O
sized	NN	O	O
alleles	NN	O	O
represent	NN	O	O
misdiagnosis	NN	O	O
,	NN	O	O
sample	NN	O	O
mix	NN	O	O
-	NN	O	O
up	NN	O	O
,	NN	O	O
or	NN	O	O
clerical	NN	O	O
error	NN	O	O
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
12	NN	O	O
patients	NN	O	O
represent	NN	O	O
possible	NN	O	O
phenocopies	NN	O	O
for	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
at	NN	O	O
least	NN	O	O
four	NN	O	O
cases	NN	O	O
,	NN	O	O
family	NN	O	O
studies	NN	O	O
of	NN	O	O
these	NN	O	O
phenocopies	NN	O	O
excluded	NN	O	O
4p16	NN	O	O
.	NN	O	O

3	NN	O	O
as	NN	O	O
the	NN	O	O
region	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
HD	NN	O	B-Disease
gene	NN	O	O
that	NN	O	O
are	NN	O	O
other	NN	O	O
than	NN	O	O
CAG	NN	O	O
expansion	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
excluded	NN	O	O
for	NN	O	O
the	NN	O	O
remaining	NN	O	O
eight	NN	O	O
cases	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
in	NN	O	O
as	NN	O	O
many	NN	O	O
as	NN	O	O
seven	NN	O	O
of	NN	O	O
these	NN	O	O
persons	NN	O	O
,	NN	O	O
retrospective	NN	O	O
review	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
clinical	NN	O	O
features	NN	O	O
identified	NN	O	O
characteristics	NN	O	O
not	NN	O	O
typical	NN	O	O
for	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

This	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
on	NN	O	O
rare	NN	O	O
occasions	NN	O	O
mutations	NN	O	O
in	NN	O	O
other	NN	O	O
,	NN	O	O
as	NN	O	O
-	NN	O	O
yet	NN	O	O
-	NN	O	O
undefined	NN	O	O
genes	NN	O	O
can	NN	O	O
present	NN	O	O
with	NN	O	O
a	NN	O	O
clinical	NN	O	O
phenotype	NN	O	O
very	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
HD	NN	O	B-Disease

Frequent	NN	O	O
detection	NN	O	O
of	NN	O	O
codon	NN	O	O
877	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	O
receptor	NN	O	O
gene	NN	O	O
in	NN	O	O
advanced	NN	O	B-Disease
prostate	NN	O	I-Disease
cancers	NN	O	I-Disease
.	NN	O	O

Prostatic	NN	O	O
tissue	NN	O	O
specimens	NN	O	O
derived	NN	O	O
from	NN	O	O
transurethral	NN	O	O
resections	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
metastatic	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
hormone	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
androgen	NN	O	O
receptor	NN	O	O
(	NN	O	O
AR	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
-	NN	O	O
derived	NN	O	O
DNAs	NN	O	O
of	NN	O	O
6	NN	O	O
of	NN	O	O
24	NN	O	O
specimens	NN	O	O
revealed	NN	O	O
a	NN	O	O
codon	NN	O	O
877	NN	O	O
mutation	NN	O	O
(	NN	O	O
ACT	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
GCT	NN	O	O
,	NN	O	O
Thr	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Ala	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
hormone	NN	O	O
-	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
AR	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
same	NN	O	O
AR	NN	O	O
mutation	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
in	NN	O	O
a	NN	O	O
metastatic	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
cell	NN	O	O
line	NN	O	O
,	NN	O	O
LNCaP	NN	O	O
,	NN	O	O
where	NN	O	O
this	NN	O	O
mutation	NN	O	O
confers	NN	O	O
upon	NN	O	O
the	NN	O	O
AR	NN	O	O
an	NN	O	O
altered	NN	O	O
ligand	NN	O	O
-	NN	O	O
binding	NN	O	O
specificity	NN	O	O
which	NN	O	O
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
estrogens	NN	O	O
,	NN	O	O
progestagens	NN	O	O
,	NN	O	O
and	NN	O	O
antiandrogens	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
analogous	NN	O	O
to	NN	O	O
an	NN	O	O
activated	NN	O	O
/	NN	O	O
altered	NN	O	O
growth	NN	O	O
factor	NN	O	O
receptor	NN	O	O
oncogene	NN	O	O
,	NN	O	O
codon	NN	O	O
877	NN	O	O
mutant	NN	O	O
AR	NN	O	O
with	NN	O	O
altered	NN	O	O
ligand	NN	O	O
binding	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
selective	NN	O	O
growth	NN	O	O
advantage	NN	O	O
in	NN	O	O
the	NN	O	O
genesis	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
advanced	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
estrogens	NN	O	O
are	NN	O	O
used	NN	O	O
infrequently	NN	O	O
,	NN	O	O
antiandrogens	NN	O	O
are	NN	O	O
used	NN	O	O
increasingly	NN	O	O
in	NN	O	O
hormonal	NN	O	O
therapy	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	B-Disease
prostate	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
these	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
on	NN	O	O
the	NN	O	O
codon	NN	O	O
877	NN	O	O
mutant	NN	O	O
AR	NN	O	O
further	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
frequently	NN	O	O
observed	NN	O	O
AR	NN	O	O
mutation	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
treatment	NN	O	O
refractory	NN	O	O
disease	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
gene	NN	O	O
for	NN	O	O
alkaptonuria	NN	O	B-Disease
(	NN	O	O
AKU	NN	O	B-Disease
)	NN	O	O
maps	NN	O	O
to	NN	O	O
chromosome	NN	O	O
3q	NN	O	O
.	NN	O	O

Alkaptonuria	NN	O	B-Disease
(	NN	O	O
AKU	NN	O	B-Disease
;	NN	O	O
McKusick	NN	O	O
no	NN	O	O
.	NN	O	O
203500	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
homogentisic	NN	O	O
acid	NN	O	O
oxidase	NN	O	O
activity	NN	O	O
.	NN	O	O

Patients	NN	O	O
excrete	NN	O	O
large	NN	O	O
amounts	NN	O	O
of	NN	O	O
homogentisic	NN	O	O
acid	NN	O	O
in	NN	O	O
their	NN	O	O
urine	NN	O	O
and	NN	O	O
a	NN	O	O
black	NN	O	O
ochronotic	NN	O	O
pigment	NN	O	O
is	NN	O	O
deposited	NN	O	O
in	NN	O	O
their	NN	O	O
cartilage	NN	O	O
and	NN	O	O
collagenous	NN	O	O
tissues	NN	O	O
.	NN	O	O

Ochronosis	NN	O	B-Disease
is	NN	O	O
the	NN	O	O
predominant	NN	O	O
clinical	NN	O	O
complication	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
leading	NN	O	O
to	NN	O	O
ochronotic	NN	O	B-Disease
arthropathy	NN	O	I-Disease
,	NN	O	O
dark	NN	O	O
urine	NN	O	O
,	NN	O	O
pigment	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
skin	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
clinical	NN	O	O
features	NN	O	O
.	NN	O	O

A	NN	O	O
mutation	NN	O	O
causing	NN	O	O
alkaptonuria	NN	O	B-Disease
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
has	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	O
16	NN	O	O
.	NN	O	O

Considering	NN	O	O
conserved	NN	O	O
synteny	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
map	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
to	NN	O	O
chromosome	NN	O	O
3q	NN	O	O
in	NN	O	O
six	NN	O	O
alkaptonuria	NN	O	B-Disease
pedigrees	NN	O	O
of	NN	O	O
Slovak	NN	O	O
origin	NN	O	O
.	NN	O	O
.	NN	O	O

Structure	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
gene	NN	O	O
SGLT1	NN	O	O
.	NN	O	O

Intestinal	NN	O	O
uptake	NN	O	O
of	NN	O	O
dietary	NN	O	O
glucose	NN	O	O
and	NN	O	O
galactose	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
SGLT1	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
of	NN	O	O
the	NN	O	O
brush	NN	O	O
border	NN	O	O
.	NN	O	O

An	NN	O	O
SGLT1	NN	O	O
missense	NN	O	O
mutation	NN	O	O
underlies	NN	O	O
hereditary	NN	O	B-Disease
glucose	NN	O	I-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
potentially	NN	O	O
fatal	NN	O	O
diarrhea	NN	O	B-Disease
;	NN	O	O
conversely	NN	O	O
,	NN	O	O
oral	NN	O	O
rehydration	NN	O	O
therapy	NN	O	O
exploits	NN	O	O
normal	NN	O	O
transport	NN	O	O
to	NN	O	O
alleviate	NN	O	O
life	NN	O	O
-	NN	O	O
threatening	NN	O	O
diarrhea	NN	O	B-Disease
of	NN	O	O
infectious	NN	O	O
origin	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
mapped	NN	O	O
the	NN	O	O
entire	NN	O	O
human	NN	O	O
SGLT1	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
gene	NN	O	O
from	NN	O	O
cosmid	NN	O	O
and	NN	O	O
lambda	NN	O	O
phage	NN	O	O
clones	NN	O	O
representing	NN	O	O
a	NN	O	O
genomic	NN	O	O
region	NN	O	O
of	NN	O	O
112	NN	O	O
kilobases	NN	O	O
.	NN	O	O

Transcription	NN	O	O
initiation	NN	O	O
occurred	NN	O	O
from	NN	O	O
a	NN	O	O
site	NN	O	O
27	NN	O	O
base	NN	O	O
pairs	NN	O	O
3	NN	O	O
of	NN	O	O
a	NN	O	O
TATAA	NN	O	O
sequence	NN	O	O
.	NN	O	O

All	NN	O	O
exon	NN	O	O
-	NN	O	O
flanking	NN	O	O
regions	NN	O	O
were	NN	O	O
sequenced	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
entire	NN	O	O
112	NN	O	O
-	NN	O	O
kilobase	NN	O	O
region	NN	O	O
mapped	NN	O	O
with	NN	O	O
four	NN	O	O
restriction	NN	O	O
enzymes	NN	O	O
.	NN	O	O

SGLT1	NN	O	O
is	NN	O	O
comprised	NN	O	O
of	NN	O	O
15	NN	O	O
exons	NN	O	O
(	NN	O	O
spanning	NN	O	O
72	NN	O	O
kilobases	NN	O	O
)	NN	O	O
;	NN	O	O
a	NN	O	O
possible	NN	O	O
evolutionary	NN	O	O
origin	NN	O	O
from	NN	O	O
a	NN	O	O
six	NN	O	O
-	NN	O	O
membrane	NN	O	O
-	NN	O	O
span	NN	O	O
ancestral	NN	O	O
precursor	NN	O	O
via	NN	O	O
a	NN	O	O
gene	NN	O	O
duplication	NN	O	O
event	NN	O	O
is	NN	O	O
suggested	NN	O	O
from	NN	O	O
comparison	NN	O	O
of	NN	O	O
exons	NN	O	O
against	NN	O	O
protein	NN	O	O
secondary	NN	O	O
structure	NN	O	O
and	NN	O	O
from	NN	O	O
sequence	NN	O	O
considerations	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	O
1	NN	O	O
causing	NN	O	O
glucose	NN	O	B-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
is	NN	O	O
also	NN	O	O
described	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
dependent	NN	O	O
cotransporter	NN	O	O
gene	NN	O	O
structure	NN	O	O
reported	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
facilitate	NN	O	O
the	NN	O	O
search	NN	O	O
for	NN	O	O
new	NN	O	O
glucose	NN	O	B-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
-	NN	O	O
related	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
important	NN	O	O
gene	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
future	NN	O	O
evolutionary	NN	O	O
comparisons	NN	O	O
with	NN	O	O
other	NN	O	O
Na	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
dependent	NN	O	O
cotransporters	NN	O	O
.	NN	O	O
.	NN	O	O

Four	NN	O	O
novel	NN	O	O
PEPD	NN	O	O
alleles	NN	O	O
causing	NN	O	O
prolidase	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
PEPD	NN	O	O
locus	NN	O	O
cause	NN	O	O
prolidase	NN	O	B-Disease
deficiency	NN	O	I-Disease
(	NN	O	O
McKusick	NN	O	O
170100	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
rare	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disorder	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
iminodipeptiduria	NN	O	B-Disease
,	NN	O	O
skin	NN	O	B-Disease
ulcers	NN	O	I-Disease
,	NN	O	O
mental	NN	O	B-Disease
retardation	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
recurrent	NN	O	O
infections	NN	O	O
.	NN	O	O

Four	NN	O	O
PEPD	NN	O	O
mutations	NN	O	O
from	NN	O	O
five	NN	O	O
severely	NN	O	O
affected	NN	O	O
individuals	NN	O	O
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
reverse	NN	O	O
-	NN	O	O
transcribed	NN	O	O
,	NN	O	O
PCR	NN	O	O
-	NN	O	O
amplified	NN	O	O
(	NN	O	O
RT	NN	O	O
-	NN	O	O
PCR	NN	O	O
)	NN	O	O
cDNA	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
on	NN	O	O
four	NN	O	O
overlapping	NN	O	O
cDNA	NN	O	O
fragments	NN	O	O
covering	NN	O	O
the	NN	O	O
entire	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
PEPD	NN	O	O
gene	NN	O	O
and	NN	O	O
detected	NN	O	O
abnormal	NN	O	O
SSCP	NN	O	O
bands	NN	O	O
for	NN	O	O
the	NN	O	O
fragment	NN	O	O
spanning	NN	O	O
all	NN	O	O
or	NN	O	O
part	NN	O	O
of	NN	O	O
exons	NN	O	O
13	NN	O	O
-	NN	O	O
15	NN	O	O
in	NN	O	O
three	NN	O	O
of	NN	O	O
the	NN	O	O
probands	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
cDNAs	NN	O	O
showed	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
1342	NN	O	O
substitution	NN	O	O
(	NN	O	O
G448R	NN	O	O
)	NN	O	O
in	NN	O	O
two	NN	O	O
patients	NN	O	O
and	NN	O	O
a	NN	O	O
3	NN	O	O
-	NN	O	O
bp	NN	O	O
deletion	NN	O	O
(	NN	O	O
delta	NN	O	O
E452	NN	O	O
or	NN	O	O
delta	NN	O	O
E453	NN	O	O
)	NN	O	O
in	NN	O	O
another	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
other	NN	O	O
two	NN	O	O
probands	NN	O	O
the	NN	O	O
amplified	NN	O	O
products	NN	O	O
were	NN	O	O
of	NN	O	O
reduced	NN	O	O
size	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
these	NN	O	O
mutant	NN	O	O
cDNAs	NN	O	O
revealed	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
exon	NN	O	O
5	NN	O	O
in	NN	O	O
one	NN	O	O
patient	NN	O	O
and	NN	O	O
of	NN	O	O
exon	NN	O	O
7	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

Intronic	NN	O	O
sequences	NN	O	O
flanking	NN	O	O
exons	NN	O	O
5	NN	O	O
and	NN	O	O
7	NN	O	O
were	NN	O	O
identified	NN	O	O
using	NN	O	O
inverse	NN	O	O
PCR	NN	O	O
followed	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Conventional	NN	O	O
PCR	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
then	NN	O	O
established	NN	O	O
the	NN	O	O
intron	NN	O	O
-	NN	O	O
exon	NN	O	O
borders	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
revealing	NN	O	O
two	NN	O	O
splice	NN	O	O
acceptor	NN	O	O
mutations	NN	O	O
a	NN	O	O
G	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
C	NN	O	O
substitution	NN	O	O
at	NN	O	O
position	NN	O	O
-	NN	O	O
1	NN	O	O
of	NN	O	O
intron	NN	O	O
4	NN	O	O
and	NN	O	O
an	NN	O	O
A	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
G	NN	O	O
substitution	NN	O	O
at	NN	O	O
position	NN	O	O
-	NN	O	O
2	NN	O	O
of	NN	O	O
intron	NN	O	O
6	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
severe	NN	O	O
form	NN	O	O
of	NN	O	O
prolidase	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
multiple	NN	O	O
PEPD	NN	O	O
alleles	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
attempt	NN	O	O
to	NN	O	O
begin	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
describing	NN	O	O
these	NN	O	O
alleles	NN	O	O
and	NN	O	O
cataloging	NN	O	O
their	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
.	NN	O	O
.	NN	O	O

Recombinations	NN	O	O
in	NN	O	O
individuals	NN	O	O
homozygous	NN	O	O
by	NN	O	O
descent	NN	O	O
localize	NN	O	O
the	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
locus	NN	O	O
in	NN	O	O
a	NN	O	O
cloned	NN	O	O
450	NN	O	O
-	NN	O	O
kb	NN	O	O
interval	NN	O	O
.	NN	O	O

The	NN	O	O
locus	NN	O	O
for	NN	O	O
Friedreich	NN	O	B-Disease
ataxia	NN	O	I-Disease
(	NN	O	O
FRDA	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
severe	NN	O	O
neurodegenerative	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
tightly	NN	O	O
linked	NN	O	O
to	NN	O	O
markers	NN	O	O
D9S5	NN	O	O
and	NN	O	O
D9S15	NN	O	O
,	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
rare	NN	O	O
recombination	NN	O	O
events	NN	O	O
has	NN	O	O
suggested	NN	O	O
the	NN	O	O
order	NN	O	O
cen	NN	O	O
-	NN	O	O
FRDA	NN	O	O
-	NN	O	O
D9S5	NN	O	O
-	NN	O	O
D9S15	NN	O	O
-	NN	O	O
qter	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
construction	NN	O	O
of	NN	O	O
a	NN	O	O
YAC	NN	O	O
contig	NN	O	O
extending	NN	O	O
800	NN	O	O
kb	NN	O	O
centromeric	NN	O	O
to	NN	O	O
D9S5	NN	O	O
and	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
five	NN	O	O
new	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
from	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
map	NN	O	O
these	NN	O	O
markers	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
FRDA	NN	O	B-Disease
locus	NN	O	O
,	NN	O	O
all	NN	O	O
within	NN	O	O
a	NN	O	O
1	NN	O	O
-	NN	O	O
cM	NN	O	O
confidence	NN	O	O
interval	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
genetic	NN	O	O
information	NN	O	O
of	NN	O	O
available	NN	O	O
FRDA	NN	O	B-Disease
families	NN	O	O
by	NN	O	O
considering	NN	O	O
homozygosity	NN	O	O
by	NN	O	O
descent	NN	O	O
and	NN	O	O
association	NN	O	O
with	NN	O	O
founder	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
isolated	NN	O	O
populations	NN	O	O
.	NN	O	O

This	NN	O	O
approach	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
identify	NN	O	O
one	NN	O	O
phase	NN	O	O
-	NN	O	O
known	NN	O	O
recombination	NN	O	O
and	NN	O	O
one	NN	O	O
probable	NN	O	O
historic	NN	O	O
recombination	NN	O	O
on	NN	O	O
haplotypes	NN	O	O
from	NN	O	O
Reunion	NN	O	O
Island	NN	O	O
patients	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
place	NN	O	O
three	NN	O	O
of	NN	O	O
the	NN	O	O
five	NN	O	O
markers	NN	O	O
proximal	NN	O	O
to	NN	O	O
FRDA	NN	O	B-Disease
.	NN	O	O

This	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
identification	NN	O	O
of	NN	O	O
close	NN	O	O
FRDA	NN	O	B-Disease
flanking	NN	O	O
markers	NN	O	O
on	NN	O	O
the	NN	O	O
centromeric	NN	O	O
side	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
other	NN	O	O
markers	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
narrow	NN	O	O
the	NN	O	O
breakpoint	NN	O	O
of	NN	O	O
a	NN	O	O
previously	NN	O	O
identified	NN	O	O
distal	NN	O	O
recombination	NN	O	O
that	NN	O	O
is	NN	O	O
>	NN	O	O
180	NN	O	O
kb	NN	O	O
from	NN	O	O
D9S5	NN	O	O
(	NN	O	O
26P	NN	O	O
)	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
place	NN	O	O
the	NN	O	O
FRDA	NN	O	B-Disease
locus	NN	O	O
in	NN	O	O
a	NN	O	O
450	NN	O	O
-	NN	O	O
kb	NN	O	O
interval	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
small	NN	O	O
enough	NN	O	O
for	NN	O	O
direct	NN	O	O
search	NN	O	O
of	NN	O	O
candidate	NN	O	O
genes	NN	O	O
.	NN	O	O

A	NN	O	O
detailed	NN	O	O
rare	NN	O	O
cutter	NN	O	O
restriction	NN	O	O
map	NN	O	O
and	NN	O	O
a	NN	O	O
cosmid	NN	O	O
contig	NN	O	O
covering	NN	O	O
this	NN	O	O
interval	NN	O	O
were	NN	O	O
constructed	NN	O	O
and	NN	O	O
should	NN	O	O
facilitate	NN	O	O
the	NN	O	O
search	NN	O	O
of	NN	O	O
genes	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O
.	NN	O	O

Investigation	NN	O	O
of	NN	O	O
thermoregulatory	NN	O	O
characteristics	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
survey	NN	O	O
instrument	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
assess	NN	O	O
temperature	NN	O	O
regulation	NN	O	O
characteristics	NN	O	O
in	NN	O	O
children	NN	O	O
with	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
compared	NN	O	O
to	NN	O	O
3	NN	O	O
control	NN	O	O
groups	NN	O	O
sibs	NN	O	O
of	NN	O	O
PWS	NN	O	B-Disease
patients	NN	O	O
(	NN	O	O
SIB	NN	O	O
)	NN	O	O
,	NN	O	O
neurodevelopmentally	NN	O	B-Disease
handicapped	NN	O	I-Disease
children	NN	O	O
(	NN	O	O
ND	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
age	NN	O	O
and	NN	O	O
gender	NN	O	O
matched	NN	O	O
well	NN	O	O
children	NN	O	O
(	NN	O	O
WC	NN	O	O
)	NN	O	O
.	NN	O	O

Significant	NN	O	O
differences	NN	O	O
were	NN	O	O
found	NN	O	O
between	NN	O	O
PWS	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
SIB	NN	O	O
controls	NN	O	O
,	NN	O	O
and	NN	O	O
WC	NN	O	O
controls	NN	O	O
in	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
febrile	NN	O	O
convulsions	NN	O	O
,	NN	O	O
fever	NN	O	O
-	NN	O	O
associated	NN	O	O
symptoms	NN	O	O
,	NN	O	O
and	NN	O	O
temperature	NN	O	O
less	NN	O	O
than	NN	O	O
94	NN	O	O
degrees	NN	O	O
F	NN	O	O
.	NN	O	O

No	NN	O	O
differences	NN	O	O
were	NN	O	O
noted	NN	O	O
in	NN	O	O
any	NN	O	O
variable	NN	O	O
between	NN	O	O
the	NN	O	O
PWS	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
the	NN	O	O
ND	NN	O	O
controls	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
abnormalities	NN	O	O
are	NN	O	O
not	NN	O	O
unique	NN	O	O
to	NN	O	O
PWS	NN	O	B-Disease
,	NN	O	O
but	NN	O	O
can	NN	O	O
occur	NN	O	O
in	NN	O	O
any	NN	O	O
neurodevelopmentally	NN	O	B-Disease
handicapped	NN	O	I-Disease
individual	NN	O	O
,	NN	O	O
further	NN	O	O
suggesting	NN	O	O
these	NN	O	O
do	NN	O	O
not	NN	O	O
necessarily	NN	O	O
reflect	NN	O	O
syndrome	NN	O	O
-	NN	O	O
specific	NN	O	O
hypothalamic	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Phenotypic	NN	O	O
variation	NN	O	O
including	NN	O	O
retinitis	NN	O	B-Disease
pigmentosa	NN	O	I-Disease
,	NN	O	O
pattern	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
fundus	NN	O	B-Disease
flavimaculatus	NN	O	I-Disease
in	NN	O	O
a	NN	O	O
single	NN	O	O
family	NN	O	O
with	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
codon	NN	O	O
153	NN	O	O
or	NN	O	O
154	NN	O	O
of	NN	O	O
the	NN	O	O
peripherin	NN	O	O
/	NN	O	O
RDS	NN	O	O
gene	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
OBJECTIVES	NN	O	O
Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
peripherin	NN	O	O
/	NN	O	O
RDS	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
retinitis	NN	O	I-Disease
pigmentosa	NN	O	I-Disease
,	NN	O	O
pattern	NN	O	O
macular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
and	NN	O	O
retinitis	NN	O	B-Disease
punctata	NN	O	I-Disease
albescens	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
report	NN	O	O
herein	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
three	NN	O	O
separate	NN	O	O
phenotypes	NN	O	O
within	NN	O	O
a	NN	O	O
single	NN	O	O
family	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
3	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
of	NN	O	O
codon	NN	O	O
153	NN	O	O
or	NN	O	O
154	NN	O	O
of	NN	O	O
the	NN	O	O
peripherin	NN	O	O
/	NN	O	O
RDS	NN	O	O
gene	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
Case	NN	O	O
reports	NN	O	O
with	NN	O	O
clinical	NN	O	O
features	NN	O	O
,	NN	O	O
fluorescein	NN	O	O
angiography	NN	O	O
,	NN	O	O
kinetic	NN	O	O
perimetry	NN	O	O
,	NN	O	O
electrophysiological	NN	O	O
studies	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
genetics	NN	O	O
.	NN	O	O

SETTING	NN	O	O
University	NN	O	O
medical	NN	O	O
centers	NN	O	O
.	NN	O	O

PATIENTS	NN	O	O
A	NN	O	O
75	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
woman	NN	O	O
,	NN	O	O
her	NN	O	O
two	NN	O	O
daughters	NN	O	O
(	NN	O	O
aged	NN	O	O
44	NN	O	O
and	NN	O	O
50	NN	O	O
years	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
her	NN	O	O
49	NN	O	O
-	NN	O	O
year	NN	O	O
-	NN	O	O
old	NN	O	O
son	NN	O	O
were	NN	O	O
screened	NN	O	O
for	NN	O	O
peripherin	NN	O	O
/	NN	O	O
RDS	NN	O	O
mutations	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
multiple	NN	O	O
phenotypes	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
The	NN	O	O
mother	NN	O	O
presented	NN	O	O
at	NN	O	O
age	NN	O	O
63	NN	O	O
years	NN	O	O
with	NN	O	O
a	NN	O	O
profoundly	NN	O	O
abnormal	NN	O	O
electroretinogram	NN	O	O
(	NN	O	O
ERG	NN	O	O
)	NN	O	O
and	NN	O	O
adult	NN	O	O
-	NN	O	O
onset	NN	O	O
retinitis	NN	O	B-Disease
pigmentosa	NN	O	I-Disease
that	NN	O	O
progressed	NN	O	O
dramatically	NN	O	O
over	NN	O	O
12	NN	O	O
years	NN	O	O
,	NN	O	O
with	NN	O	O
marked	NN	O	O
loss	NN	O	O
of	NN	O	O
peripheral	NN	O	O
visual	NN	O	O
field	NN	O	O
.	NN	O	O

One	NN	O	O
daughter	NN	O	O
developed	NN	O	O
pattern	NN	O	B-Disease
macular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
at	NN	O	O
age	NN	O	O
31	NN	O	O
years	NN	O	O
.	NN	O	O

At	NN	O	O
age	NN	O	O
44	NN	O	O
years	NN	O	O
,	NN	O	O
her	NN	O	O
ERG	NN	O	O
was	NN	O	O
moderately	NN	O	O
abnormal	NN	O	O
but	NN	O	O
her	NN	O	O
clinical	NN	O	O
disease	NN	O	O
was	NN	O	O
limited	NN	O	O
to	NN	O	O
the	NN	O	O
macula	NN	O	O
.	NN	O	O

Another	NN	O	O
daughter	NN	O	O
presented	NN	O	O
at	NN	O	O
age	NN	O	O
42	NN	O	O
years	NN	O	O
with	NN	O	O
macular	NN	O	B-Disease
degeneration	NN	O	I-Disease
and	NN	O	O
over	NN	O	O
10	NN	O	O
years	NN	O	O
developed	NN	O	O
the	NN	O	O
clinical	NN	O	O
picture	NN	O	O
of	NN	O	O
fundus	NN	O	B-Disease
flavimaculatus	NN	O	I-Disease
.	NN	O	O

Her	NN	O	O
peripheral	NN	O	O
visual	NN	O	O
field	NN	O	O
was	NN	O	O
preserved	NN	O	O
but	NN	O	O
her	NN	O	O
ERG	NN	O	O
was	NN	O	O
moderately	NN	O	O
abnormal	NN	O	O
.	NN	O	O

The	NN	O	O
son	NN	O	O
had	NN	O	O
onset	NN	O	O
of	NN	O	O
macular	NN	O	B-Disease
degeneration	NN	O	I-Disease
at	NN	O	O
age	NN	O	O
44	NN	O	O
years	NN	O	O
.	NN	O	O

Pericentral	NN	O	B-Disease
scotomas	NN	O	I-Disease
were	NN	O	O
present	NN	O	O
and	NN	O	O
the	NN	O	O
ERG	NN	O	O
was	NN	O	O
markedly	NN	O	O
abnormal	NN	O	O
.	NN	O	O

Fluorescein	NN	O	O
angiography	NN	O	O
revealed	NN	O	O
punctate	NN	O	O
pigment	NN	O	O
epithelial	NN	O	O
transmission	NN	O	O
defects	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
A	NN	O	O
3	NN	O	O
-	NN	O	O
base	NN	O	O
pair	NN	O	O
deletion	NN	O	O
of	NN	O	O
codon	NN	O	O
153	NN	O	O
or	NN	O	O
154	NN	O	O
of	NN	O	O
the	NN	O	O
peripherin	NN	O	O
/	NN	O	O
RDS	NN	O	O
gene	NN	O	O
can	NN	O	O
produce	NN	O	O
clinically	NN	O	O
disparate	NN	O	O
phenotypes	NN	O	O
even	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
.	NN	O	O
.	NN	O	O

Assignment	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
gene	NN	O	O
SGLT1	NN	O	O
to	NN	O	O
chromosome	NN	O	O
22q13	NN	O	O
.	NN	O	O
1	NN	O	O
.	NN	O	O

The	NN	O	O
Na	NN	O	O
+	NN	O	O
/	NN	O	O
glucose	NN	O	O
cotransporter	NN	O	O
gene	NN	O	O
SGLT1	NN	O	O
encodes	NN	O	O
the	NN	O	O
primary	NN	O	O
carrier	NN	O	O
protein	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
the	NN	O	O
dietary	NN	O	O
sugars	NN	O	O
glucose	NN	O	O
and	NN	O	O
galactose	NN	O	O
from	NN	O	O
the	NN	O	O
intestinal	NN	O	O
lumen	NN	O	O
.	NN	O	O

SGLT1	NN	O	O
transport	NN	O	O
activity	NN	O	O
is	NN	O	O
currently	NN	O	O
exploited	NN	O	O
in	NN	O	O
oral	NN	O	O
rehydration	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
75	NN	O	O
-	NN	O	O
kDa	NN	O	O
glycoprotein	NN	O	O
is	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
brush	NN	O	O
border	NN	O	O
of	NN	O	O
the	NN	O	O
intestinal	NN	O	O
epithelium	NN	O	O
and	NN	O	O
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
comprise	NN	O	O
12	NN	O	O
membrane	NN	O	O
spans	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
autosomal	NN	O	B-Disease
recessive	NN	O	I-Disease
disease	NN	O	I-Disease
glucose	NN	O	I-Disease
/	NN	O	I-Disease
galactose	NN	O	I-Disease
malabsorption	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
underlying	NN	O	O
cause	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
SGLT1	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
Asp28	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Asn	NN	O	O
change	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
eliminate	NN	O	O
SGLT1	NN	O	O
transport	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
SGLT1	NN	O	O
gene	NN	O	O
was	NN	O	O
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
reside	NN	O	O
on	NN	O	O
the	NN	O	O
distal	NN	O	O
q	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22	NN	O	O
(	NN	O	O
11	NN	O	O
.	NN	O	O
2	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
qter	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
cosmid	NN	O	O
probe	NN	O	O
for	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
which	NN	O	O
refines	NN	O	O
the	NN	O	O
localization	NN	O	O
to	NN	O	O
22q13	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
and	NN	O	O
provide	NN	O	O
an	NN	O	O
example	NN	O	O
of	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
the	NN	O	O
SGLT1	NN	O	O
probe	NN	O	O
as	NN	O	O
a	NN	O	O
diagnostic	NN	O	O
for	NN	O	O
genetic	NN	O	B-Disease
diseases	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
translocations	NN	O	O
of	NN	O	O
chromosome	NN	O	O
22	NN	O	O
.	NN	O	O

Restriction	NN	O	O
of	NN	O	O
ocular	NN	O	O
fundus	NN	O	O
lesions	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
subgroup	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
in	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
patients	NN	O	O
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
APC	NN	O	B-Disease
,	NN	O	O
causes	NN	O	O
adenomatous	NN	O	B-Disease
polyposis	NN	O	I-Disease
coli	NN	O	I-Disease
,	NN	O	O
a	NN	O	O
condition	NN	O	O
causing	NN	O	O
predisposition	NN	O	O
to	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
syndrome	NN	O	O
inconsistently	NN	O	O
associates	NN	O	O
characteristic	NN	O	O
patches	NN	O	O
of	NN	O	O
congenital	NN	O	B-Disease
hypertrophy	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
retinal	NN	O	I-Disease
pigment	NN	O	I-Disease
epithelium	NN	O	I-Disease
(	NN	O	O
CHRPE	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Ocular	NN	O	O
examination	NN	O	O
revealed	NN	O	O
that	NN	O	O
patients	NN	O	O
expressing	NN	O	O
CHRPE	NN	O	B-Disease
tend	NN	O	O
to	NN	O	O
cluster	NN	O	O
within	NN	O	O
specific	NN	O	O
families	NN	O	O
.	NN	O	O

The	NN	O	O
exact	NN	O	O
APC	NN	O	B-Disease
mutation	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
42	NN	O	O
unrelated	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
cases	NN	O	O
these	NN	O	O
mutations	NN	O	O
were	NN	O	O
predicted	NN	O	O
to	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
extent	NN	O	O
of	NN	O	O
CHRPE	NN	O	B-Disease
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
along	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
.	NN	O	O

CHRPE	NN	O	B-Disease
lesions	NN	O	O
are	NN	O	O
almost	NN	O	O
always	NN	O	O
absent	NN	O	O
if	NN	O	O
the	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
before	NN	O	O
exon	NN	O	O
9	NN	O	O
,	NN	O	O
but	NN	O	O
are	NN	O	O
systematically	NN	O	O
present	NN	O	O
if	NN	O	O
it	NN	O	O
occurs	NN	O	O
after	NN	O	O
this	NN	O	O
exon	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
observed	NN	O	O
among	NN	O	O
affected	NN	O	O
patients	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
part	NN	O	O
from	NN	O	O
different	NN	O	O
allelic	NN	O	O
manifestations	NN	O	O
of	NN	O	O
APC	NN	O	B-Disease
mutations	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
mutations	NN	O	O
on	NN	O	O
the	NN	O	O
ERG	NN	O	O
in	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

PURPOSE	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
earlier	NN	O	O
findings	NN	O	O
of	NN	O	O
a	NN	O	O
negative	NN	O	O
electroretinogram	NN	O	O
(	NN	O	O
ERG	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
led	NN	O	O
them	NN	O	O
to	NN	O	O
investigate	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
deletions	NN	O	O
and	NN	O	O
ERGs	NN	O	O
in	NN	O	O
five	NN	O	O
boys	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
authors	NN	O	O
wanted	NN	O	O
to	NN	O	O
determined	NN	O	O
whether	NN	O	O
there	NN	O	O
were	NN	O	O
similar	NN	O	O
ERG	NN	O	O
findings	NN	O	O
in	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
mdx	NN	O	O
mouse	NN	O	O
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

Ganzfeld	NN	O	O
ERGs	NN	O	O
were	NN	O	O
recorded	NN	O	O
in	NN	O	O
five	NN	O	O
boys	NN	O	O
with	NN	O	O
DMD	NN	O	B-Disease
after	NN	O	O
a	NN	O	O
complete	NN	O	O
ophthalmic	NN	O	O
examination	NN	O	O
.	NN	O	O

The	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
.	NN	O	O

ERGs	NN	O	O
were	NN	O	O
recorded	NN	O	O
in	NN	O	O
anesthetized	NN	O	O
mdx	NN	O	O
and	NN	O	O
control	NN	O	O
mice	NN	O	O
with	NN	O	O
a	NN	O	O
modified	NN	O	O
Grass	NN	O	O
photostimulator	NN	O	O
(	NN	O	O
Grass	NN	O	O
Instrument	NN	O	O
Company	NN	O	O
,	NN	O	O
Quincy	NN	O	O
,	NN	O	O
MA	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Ophthalmic	NN	O	O
examinations	NN	O	O
in	NN	O	O
all	NN	O	O
five	NN	O	O
boys	NN	O	O
had	NN	O	O
normal	NN	O	O
findings	NN	O	O
,	NN	O	O
yet	NN	O	O
an	NN	O	O
abnormal	NN	O	O
negative	NN	O	O
ERG	NN	O	O
was	NN	O	O
recorded	NN	O	O
for	NN	O	O
each	NN	O	O
subject	NN	O	O
.	NN	O	O

The	NN	O	O
subjects	NN	O	O
gene	NN	O	O
deletions	NN	O	O
were	NN	O	O
variable	NN	O	O
,	NN	O	O
ranging	NN	O	O
from	NN	O	O
large	NN	O	O
deletions	NN	O	O
to	NN	O	O
no	NN	O	O
detectable	NN	O	O
deletions	NN	O	O
.	NN	O	O

The	NN	O	O
ERGs	NN	O	O
of	NN	O	O
the	NN	O	O
mdx	NN	O	O
mice	NN	O	O
were	NN	O	O
normal	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
believe	NN	O	O
the	NN	O	O
unique	NN	O	O
ERG	NN	O	O
recorded	NN	O	O
for	NN	O	O
the	NN	O	O
human	NN	O	O
subjects	NN	O	O
is	NN	O	O
a	NN	O	O
manifestation	NN	O	O
of	NN	O	O
DMD	NN	O	B-Disease
associated	NN	O	O
with	NN	O	O
defects	NN	O	O
at	NN	O	O
the	NN	O	O
dystrophin	NN	O	O
gene	NN	O	O
locus	NN	O	O
and	NN	O	O
represents	NN	O	O
a	NN	O	O
new	NN	O	O
clinical	NN	O	O
entity	NN	O	O
.	NN	O	O

The	NN	O	O
ERG	NN	O	O
of	NN	O	O
the	NN	O	O
mdx	NN	O	O
mouse	NN	O	O
may	NN	O	O
be	NN	O	O
spared	NN	O	O
for	NN	O	O
several	NN	O	O
reasons	NN	O	O
,	NN	O	O
including	NN	O	O
milder	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
gene	NN	O	O
defect	NN	O	O
,	NN	O	O
differences	NN	O	O
in	NN	O	O
muscle	NN	O	O
and	NN	O	O
retinal	NN	O	O
gene	NN	O	O
product	NN	O	O
,	NN	O	O
or	NN	O	O
species	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
biochemical	NN	O	O
role	NN	O	O
of	NN	O	O
dystrophin	NN	O	O
.	NN	O	O

The	NN	O	O
ERG	NN	O	O
shows	NN	O	O
promise	NN	O	O
of	NN	O	O
becoming	NN	O	O
a	NN	O	O
noninvasive	NN	O	O
diagnostic	NN	O	O
tool	NN	O	O
for	NN	O	O
DMD	NN	O	B-Disease
and	NN	O	O
its	NN	O	O
milder	NN	O	O
allelic	NN	O	O
forms	NN	O	O
.	NN	O	O
.	NN	O	O

Association	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
tumor	NN	O	I-Disease
suppressor	NN	O	O
protein	NN	O	O
with	NN	O	O
catenins	NN	O	O
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
APC	NN	O	O
appear	NN	O	O
to	NN	O	O
initiate	NN	O	O
sporadic	NN	O	O
and	NN	O	O
inherited	NN	O	O
forms	NN	O	O
of	NN	O	O
human	NN	O	O
colorectal	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Although	NN	O	O
these	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
APC	NN	O	B-Disease
gene	NN	O	O
product	NN	O	O
.	NN	O	O

Two	NN	O	O
cellular	NN	O	O
proteins	NN	O	O
that	NN	O	O
associate	NN	O	O
with	NN	O	O
APC	NN	O	B-Disease
were	NN	O	O
identified	NN	O	O
by	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
and	NN	O	O
peptide	NN	O	O
mapping	NN	O	O
as	NN	O	O
the	NN	O	O
E	NN	O	O
-	NN	O	O
cadherin	NN	O	O
-	NN	O	O
associated	NN	O	O
proteins	NN	O	O
alpha	NN	O	O
-	NN	O	O
and	NN	O	O
beta	NN	O	O
-	NN	O	O
catenin	NN	O	O
.	NN	O	O

A	NN	O	O
27	NN	O	O
-	NN	O	O
residue	NN	O	O
fragment	NN	O	O
of	NN	O	O
APC	NN	O	O
containing	NN	O	O
a	NN	O	O
15	NN	O	O
-	NN	O	O
amino	NN	O	O
acid	NN	O	O
repeat	NN	O	O
was	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
catenins	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
an	NN	O	O
important	NN	O	O
link	NN	O	O
between	NN	O	O
tumor	NN	O	B-Disease
initiation	NN	O	O
and	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
.	NN	O	O
.	NN	O	O

Difference	NN	O	O
in	NN	O	O
methylation	NN	O	O
patterns	NN	O	O
within	NN	O	O
the	NN	O	O
D15S9	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
in	NN	O	O
first	NN	O	O
cousins	NN	O	O
with	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Abnormalities	NN	O	O
of	NN	O	O
chromosome	NN	O	O
region	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
Angelman	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
AS	NN	O	B-Disease
)	NN	O	O
and	NN	O	O
Prader	NN	O	B-Disease
-	NN	O	I-Disease
Willi	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
PWS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Differences	NN	O	O
between	NN	O	O
the	NN	O	O
methylation	NN	O	O
patterns	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
chromosome	NN	O	O
15q11	NN	O	O
-	NN	O	O
13	NN	O	O
which	NN	O	O
hybridizes	NN	O	O
to	NN	O	O
the	NN	O	O
highly	NN	O	O
conserved	NN	O	O
DNA	NN	O	O
,	NN	O	O
DN34	NN	O	O
,	NN	O	O
in	NN	O	O
normal	NN	O	O
individuals	NN	O	O
and	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
AS	NN	O	B-Disease
and	NN	O	O
PWS	NN	O	B-Disease
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
first	NN	O	O
cousins	NN	O	O
are	NN	O	O
affected	NN	O	O
by	NN	O	O
AS	NN	O	B-Disease
and	NN	O	O
PWS	NN	O	B-Disease
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
familial	NN	O	O
paracentric	NN	O	O
inversion	NN	O	O
of	NN	O	O
15q11	NN	O	O
-	NN	O	O
q13	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
studies	NN	O	O
on	NN	O	O
this	NN	O	O
family	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
methylation	NN	O	O
patterns	NN	O	O
in	NN	O	O
the	NN	O	O
2	NN	O	O
conditions	NN	O	O
and	NN	O	O
the	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
genomic	NN	O	O
imprinting	NN	O	O
,	NN	O	O
whereby	NN	O	O
genetic	NN	O	O
information	NN	O	O
is	NN	O	O
expressed	NN	O	O
differently	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
parent	NN	O	O
of	NN	O	O
origin	NN	O	O
.	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
studies	NN	O	O
in	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
51	NN	O	O
families	NN	O	O
with	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
DNA	NN	O	O
haplotypes	NN	O	O
of	NN	O	O
dinucleotide	NN	O	O
repeat	NN	O	O
polymorphisms	NN	O	O
(	NN	O	O
CA	NN	O	O
repeats	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
13q14	NN	O	O
.	NN	O	O

3	NN	O	O
region	NN	O	O
,	NN	O	O
to	NN	O	O
examine	NN	O	O
these	NN	O	O
markers	NN	O	O
for	NN	O	O
association	NN	O	O
with	NN	O	O
the	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
(	NN	O	O
WND	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
marker	NN	O	O
(	NN	O	O
D13S133	NN	O	O
)	NN	O	O
described	NN	O	O
elsewhere	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
three	NN	O	O
new	NN	O	O
highly	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
(	NN	O	O
D13S314	NN	O	O
,	NN	O	O
D13S315	NN	O	O
,	NN	O	O
and	NN	O	O
D13S316	NN	O	O
)	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
WND	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
marker	NN	O	O
alleles	NN	O	O
at	NN	O	O
the	NN	O	O
loci	NN	O	O
studied	NN	O	O
and	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
D13S314	NN	O	O
,	NN	O	O
D13S133	NN	O	O
,	NN	O	O
and	NN	O	O
D13S316	NN	O	O
each	NN	O	O
show	NN	O	O
nonrandom	NN	O	O
distribution	NN	O	O
on	NN	O	O
chromosomes	NN	O	O
carrying	NN	O	O
the	NN	O	O
WND	NN	O	B-Disease
mutation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
haplotypes	NN	O	O
of	NN	O	O
these	NN	O	O
three	NN	O	O
markers	NN	O	O
and	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
highly	NN	O	O
significant	NN	O	O
differences	NN	O	O
between	NN	O	O
WND	NN	O	B-Disease
and	NN	O	O
normal	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
northern	NN	O	O
European	NN	O	O
families	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
mutation	NN	O	O
detection	NN	O	O
and	NN	O	O
molecular	NN	O	O
diagnosis	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
Wilson	NN	O	B-Disease
disease	NN	O	I-Disease
.	NN	O	O

Genetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
region	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
breast	NN	O	O
/	NN	O	O
ovarian	NN	O	O
family	NN	O	O
:	NN	O	O
refinement	NN	O	O
of	NN	O	O
the	NN	O	O
minimal	NN	O	O
region	NN	O	O
containing	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
a	NN	O	O
single	NN	O	O
multi	NN	O	O
-	NN	O	O
affected	NN	O	O
breast	NN	O	B-Disease
/	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
pedigree	NN	O	O
(	NN	O	O
BOV3	NN	O	O
)	NN	O	O
and	NN	O	O
have	NN	O	O
shown	NN	O	O
consistent	NN	O	O
inheritance	NN	O	O
of	NN	O	O
markers	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
confirming	NN	O	O
that	NN	O	O
this	NN	O	O
family	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
17q	NN	O	O
haplotypes	NN	O	O
shows	NN	O	O
a	NN	O	O
recombination	NN	O	O
event	NN	O	O
in	NN	O	O
a	NN	O	O
bilateral	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
case	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
lies	NN	O	O
distal	NN	O	O
to	NN	O	O
D17S857	NN	O	O
;	NN	O	O
D17S857	NN	O	O
is	NN	O	O
thus	NN	O	O
the	NN	O	O
new	NN	O	O
proximal	NN	O	O
boundary	NN	O	O
for	NN	O	O
the	NN	O	O
region	NN	O	O
containing	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Combining	NN	O	O
this	NN	O	O
information	NN	O	O
with	NN	O	O
previously	NN	O	O
published	NN	O	O
mapping	NN	O	O
information	NN	O	O
suggests	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
is	NN	O	O
contained	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
estimated	NN	O	O
at	NN	O	O
1	NN	O	O
-	NN	O	O
1	NN	O	O
.	NN	O	O

5	NN	O	O
Mb	NN	O	O
in	NN	O	O
length	NN	O	O
.	NN	O	O

All	NN	O	O
seven	NN	O	O
breast	NN	O	B-Disease
tumour	NN	O	I-Disease
/	NN	O	O
blood	NN	O	O
pairs	NN	O	O
examined	NN	O	O
from	NN	O	O
this	NN	O	O
family	NN	O	O
show	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
in	NN	O	O
the	NN	O	O
tumours	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
allel	NN	O	O
retained	NN	O	O
in	NN	O	O
each	NN	O	O
tumour	NN	O	B-Disease
was	NN	O	O
from	NN	O	O
the	NN	O	O
disease	NN	O	O
-	NN	O	O
bearing	NN	O	O
chromosome	NN	O	O
implicating	NN	O	O
BRCA1	NN	O	O
as	NN	O	O
a	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
sequenced	NN	O	O
the	NN	O	O
17	NN	O	O
beta	NN	O	O
-	NN	O	O
oestradiol	NN	O	O
dehydrogenase	NN	O	O
genes	NN	O	O
(	NN	O	O
EDH17B1	NN	O	O
and	NN	O	O
EDH17B2	NN	O	O
)	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
as	NN	O	O
candidate	NN	O	O
genes	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
four	NN	O	O
members	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
.	NN	O	O

No	NN	O	O
germline	NN	O	O
mutations	NN	O	O
were	NN	O	O
detected	NN	O	O
.	NN	O	O

Myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
kinase	NN	O	O
is	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
neuromuscular	NN	O	O
junctions	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
manifestation	NN	O	O
of	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
correlated	NN	O	O
to	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
expansion	NN	O	O
of	NN	O	O
an	NN	O	O
unstable	NN	O	O
[	NN	O	O
CTG	NN	O	O
]	NN	O	O
n	NN	O	O
DNA	NN	O	O
motif	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
trinucleotide	NN	O	O
motif	NN	O	O
forms	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
last	NN	O	O
,	NN	O	O
3	NN	O	O
untranslated	NN	O	O
exon	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
which	NN	O	O
potentially	NN	O	O
encodes	NN	O	O
multiple	NN	O	O
protein	NN	O	O
isoforms	NN	O	O
of	NN	O	O
a	NN	O	O
serine	NN	O	O
/	NN	O	O
threonine	NN	O	O
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
protein	NN	O	O
kinase	NN	O	O
,	NN	O	O
DM	NN	O	O
-	NN	O	O
PK	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
on	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
antisera	NN	O	O
against	NN	O	O
synthetic	NN	O	O
DM	NN	O	O
-	NN	O	O
PK	NN	O	O
peptide	NN	O	O
antigens	NN	O	O
and	NN	O	O
their	NN	O	O
use	NN	O	O
in	NN	O	O
biochemical	NN	O	O
and	NN	O	O
histochemical	NN	O	O
studies	NN	O	O
.	NN	O	O

Immunoreactive	NN	O	O
DM	NN	O	O
-	NN	O	O
kinase	NN	O	O
protein	NN	O	O
of	NN	O	O
53	NN	O	O
kD	NN	O	O
is	NN	O	O
present	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
in	NN	O	O
skeletal	NN	O	O
and	NN	O	O
cardiac	NN	O	O
muscle	NN	O	O
extracts	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
staining	NN	O	O
revealed	NN	O	O
that	NN	O	O
DM	NN	O	O
-	NN	O	O
PK	NN	O	O
is	NN	O	O
localised	NN	O	O
prominently	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
neuromuscular	NN	O	O
and	NN	O	O
myotendinous	NN	O	O
junctions	NN	O	O
(	NN	O	O
NMJs	NN	O	O
and	NN	O	O
MTJs	NN	O	O
)	NN	O	O
of	NN	O	O
human	NN	O	O
and	NN	O	O
rodent	NN	O	O
skeletal	NN	O	O
muscles	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
immunoreactive	NN	O	O
DM	NN	O	O
-	NN	O	O
PK	NN	O	O
protein	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
sarcoplasm	NN	O	O
of	NN	O	O
predominantly	NN	O	O
type	NN	O	O
I	NN	O	O
fibres	NN	O	O
in	NN	O	O
various	NN	O	O
muscles	NN	O	O
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
can	NN	O	O
also	NN	O	O
be	NN	O	O
demonstrated	NN	O	O
for	NN	O	O
NMJs	NN	O	O
of	NN	O	O
muscular	NN	O	O
tissues	NN	O	O
of	NN	O	O
adult	NN	O	O
and	NN	O	O
congenital	NN	O	O
cases	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
,	NN	O	O
with	NN	O	O
no	NN	O	O
gross	NN	O	O
changes	NN	O	O
in	NN	O	O
structural	NN	O	O
organisation	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
provide	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
further	NN	O	O
characterisation	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
kinase	NN	O	O
in	NN	O	O
protein	NN	O	O
assembly	NN	O	O
processes	NN	O	O
or	NN	O	O
signal	NN	O	O
mediation	NN	O	O
at	NN	O	O
synaptic	NN	O	O
sites	NN	O	O
and	NN	O	O
ultimately	NN	O	O
for	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
DM	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Markedly	NN	O	O
accelerated	NN	O	O
catabolism	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
(	NN	O	O
ApoA	NN	O	O
-	NN	O	O
II	NN	O	O
)	NN	O	O
and	NN	O	O
high	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
containing	NN	O	O
ApoA	NN	O	O
-	NN	O	O
II	NN	O	O
in	NN	O	O
classic	NN	O	O
lecithin	NN	O	O
:	NN	O	O
cholesterol	NN	O	B-Disease
acyltransferase	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
fish	NN	O	B-Disease
-	NN	O	I-Disease
eye	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Classic	NN	O	B-Disease
(	NN	O	I-Disease
complete	NN	O	I-Disease
)	NN	O	I-Disease
lecithin	NN	O	I-Disease
cholesterol	NN	O	I-Disease
acyltransferase	NN	O	I-Disease
(	NN	O	I-Disease
LCAT	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
Fish	NN	O	B-Disease
-	NN	O	I-Disease
eye	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
partial	NN	O	B-Disease
LCAT	NN	O	I-Disease
deficiency	NN	O	I-Disease
)	NN	O	O
are	NN	O	O
genetic	NN	O	O
syndromes	NN	O	O
associated	NN	O	O
with	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
levels	NN	O	O
of	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
cholesterol	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
atherosclerotic	NN	O	B-Disease
cardiovascular	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
metabolism	NN	O	O
of	NN	O	O
the	NN	O	O
HDL	NN	O	O
apolipoproteins	NN	O	O
(	NN	O	O
apo	NN	O	O
)	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
and	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
in	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
five	NN	O	O
patients	NN	O	O
with	NN	O	O
LCAT	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
one	NN	O	O
with	NN	O	O
classic	NN	O	B-Disease
LCAT	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
four	NN	O	O
with	NN	O	O
Fish	NN	O	B-Disease
-	NN	O	I-Disease
eye	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Plasma	NN	O	O
levels	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
were	NN	O	O
decreased	NN	O	O
to	NN	O	O
a	NN	O	O
proportionately	NN	O	O
greater	NN	O	O
extent	NN	O	O
(	NN	O	O
23	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
)	NN	O	O
than	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
(	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
plasma	NN	O	O
concentrations	NN	O	O
of	NN	O	O
HDL	NN	O	O
particles	NN	O	O
containing	NN	O	O
both	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
and	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
(	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
)	NN	O	O
were	NN	O	O
much	NN	O	O
lower	NN	O	O
(	NN	O	O
18	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
)	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
particles	NN	O	O
containing	NN	O	O
only	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
(	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
)	NN	O	O
(	NN	O	O
51	NN	O	O
%	NN	O	O
of	NN	O	O
normal	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
metabolic	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
and	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
all	NN	O	O
five	NN	O	O
patients	NN	O	O
using	NN	O	O
both	NN	O	O
exogenous	NN	O	O
radiotracer	NN	O	O
and	NN	O	O
endogenous	NN	O	O
stable	NN	O	O
isotope	NN	O	O
labeling	NN	O	O
techniques	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
plasma	NN	O	O
residence	NN	O	O
time	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
was	NN	O	O
decreased	NN	O	O
at	NN	O	O
2	NN	O	O
.	NN	O	O

08	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

27	NN	O	O
d	NN	O	O
(	NN	O	O
controls	NN	O	O
4	NN	O	O
.	NN	O	O
74	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
65	NN	O	O
days	NN	O	O
)	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
residence	NN	O	O
time	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
was	NN	O	O
even	NN	O	O
shorter	NN	O	O
at	NN	O	O
1	NN	O	O
.	NN	O	O

66	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O

24	NN	O	O
d	NN	O	O
(	NN	O	O
controls	NN	O	O
5	NN	O	O
.	NN	O	O
25	NN	O	O
+	NN	O	O
/	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
61	NN	O	O
d	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
catabolism	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
in	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
was	NN	O	O
substantially	NN	O	O
faster	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O
in	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
genetic	NN	O	O
syndromes	NN	O	O
of	NN	O	O
either	NN	O	O
complete	NN	O	B-Disease
or	NN	O	I-Disease
partial	NN	O	I-Disease
LCAT	NN	O	I-Disease
deficiency	NN	O	I-Disease
result	NN	O	O
in	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
HDL	NN	O	O
through	NN	O	O
preferential	NN	O	O
hypercatabolism	NN	O	O
of	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
and	NN	O	O
HDL	NN	O	O
particles	NN	O	O
containing	NN	O	O
apoA	NN	O	O
-	NN	O	O
II	NN	O	O
.	NN	O	O

Because	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
protective	NN	O	O
than	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
against	NN	O	O
atherosclerosis	NN	O	B-Disease
,	NN	O	O
this	NN	O	O
selective	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
metabolism	NN	O	O
of	NN	O	O
LpA	NN	O	O
-	NN	O	O
I	NN	O	O
A	NN	O	O
-	NN	O	O
II	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
potential	NN	O	O
explanation	NN	O	O
why	NN	O	O
patients	NN	O	O
with	NN	O	O
classic	NN	O	B-Disease
LCAT	NN	O	I-Disease
deficiency	NN	O	I-Disease
and	NN	O	O
Fish	NN	O	B-Disease
-	NN	O	I-Disease
eye	NN	O	I-Disease
disease	NN	O	I-Disease
are	NN	O	O
not	NN	O	O
at	NN	O	O
increased	NN	O	O
risk	NN	O	O
for	NN	O	O
premature	NN	O	O
atherosclerosis	NN	O	B-Disease
despite	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
HDL	NN	O	O
cholesterol	NN	O	O
and	NN	O	O
apoA	NN	O	O
-	NN	O	O
I	NN	O	O

X	NN	O	B-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
Saudi	NN	O	O
Arab	NN	O	O
boy	NN	O	O
presented	NN	O	O
in	NN	O	O
early	NN	O	O
childhood	NN	O	O
with	NN	O	O
thrombocytopenia	NN	O	B-Disease
,	NN	O	O
morphologically	NN	O	O
large	NN	O	O
and	NN	O	O
normal	NN	O	O
sized	NN	O	O
platelets	NN	O	O
,	NN	O	O
increased	NN	O	O
mean	NN	O	O
platelet	NN	O	O
volume	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
hypermegakaryocytic	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
clinical	NN	O	O
and	NN	O	O
laboratory	NN	O	O
evidence	NN	O	O
of	NN	O	O
any	NN	O	O
significant	NN	O	O
immunological	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

Similar	NN	O	O
findings	NN	O	O
in	NN	O	O
two	NN	O	O
other	NN	O	O
brothers	NN	O	O
suggested	NN	O	O
strongly	NN	O	O
that	NN	O	O
they	NN	O	O
were	NN	O	O
all	NN	O	O
suffering	NN	O	O
from	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
thrombocytopenic	NN	O	I-Disease
disorder	NN	O	I-Disease
.	NN	O	O

Results	NN	O	O
of	NN	O	O
DNA	NN	O	O
analysis	NN	O	O
with	NN	O	O
the	NN	O	O
probe	NN	O	O
M27	NN	O	O
beta	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
X	NN	O	O
linkage	NN	O	O
and	NN	O	O
indicate	NN	O	O
also	NN	O	O
that	NN	O	O
the	NN	O	O
locus	NN	O	O
of	NN	O	O
the	NN	O	O
relevant	NN	O	O
gene	NN	O	O
lies	NN	O	O
close	NN	O	O
to	NN	O	O
or	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
locus	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
various	NN	O	O
features	NN	O	O
which	NN	O	O
include	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
large	NN	O	O
and	NN	O	O
normal	NN	O	O
sized	NN	O	O
platelets	NN	O	O
(	NN	O	O
rather	NN	O	O
than	NN	O	O
small	NN	O	O
platelets	NN	O	O
)	NN	O	O
and	NN	O	O
freedom	NN	O	O
from	NN	O	O
significant	NN	O	O
immune	NN	O	B-Disease
deficiencies	NN	O	I-Disease
,	NN	O	O
it	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
linked	NN	O	I-Disease
recessive	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
in	NN	O	O
this	NN	O	O
family	NN	O	O
is	NN	O	O
an	NN	O	O
isolated	NN	O	O
entity	NN	O	O
quite	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
classical	NN	O	O
WAS	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
modified	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
gene	NN	O	O
producing	NN	O	O
a	NN	O	O
mild	NN	O	O
phenotypic	NN	O	O
variant	NN	O	O
cannot	NN	O	O
be	NN	O	O
excluded	NN	O	O
entirely	NN	O	O
.	NN	O	O
.	NN	O	O

Macular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
at	NN	O	O
codon	NN	O	O
172	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
slow	NN	O	O
gene	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
Recently	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
retinal	NN	O	B-Disease
degeneration	NN	O	I-Disease
slow	NN	O	O
(	NN	O	O
rds	NN	O	O
)	NN	O	O
gene	NN	O	O
which	NN	O	O
codes	NN	O	O
for	NN	O	O
peripherin	NN	O	O
-	NN	O	O
rds	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
retinitis	NN	O	I-Disease
pigmentosa	NN	O	I-Disease
.	NN	O	O

Because	NN	O	O
this	NN	O	O
gene	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
both	NN	O	O
rods	NN	O	O
and	NN	O	O
cones	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
rds	NN	O	O
gene	NN	O	O
might	NN	O	O
be	NN	O	O
expected	NN	O	O
to	NN	O	O
cause	NN	O	O
degeneration	NN	O	O
affecting	NN	O	O
either	NN	O	O
the	NN	O	O
scotopic	NN	O	O
or	NN	O	O
photopic	NN	O	O
systems	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
codon	NN	O	O
172	NN	O	O
of	NN	O	O
the	NN	O	O
rds	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
three	NN	O	O
families	NN	O	O
with	NN	O	O
autosomal	NN	O	O
dominantly	NN	O	O
inherited	NN	O	O
,	NN	O	O
progressive	NN	O	O
macular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

METHODS	NN	O	O
Affected	NN	O	O
individuals	NN	O	O
underwent	NN	O	O
ophthalmic	NN	O	O
examination	NN	O	O
,	NN	O	O
scotopic	NN	O	O
perimetry	NN	O	O
,	NN	O	O
dark	NN	O	O
adaptometry	NN	O	O
,	NN	O	O
measurement	NN	O	O
of	NN	O	O
color	NN	O	O
-	NN	O	O
contrast	NN	O	O
sensitivity	NN	O	O
,	NN	O	O
and	NN	O	O
electroretinography	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
photoreceptor	NN	O	O
dysfunction	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
In	NN	O	O
all	NN	O	O
but	NN	O	O
one	NN	O	O
affected	NN	O	O
member	NN	O	O
,	NN	O	O
symptoms	NN	O	O
of	NN	O	O
progressive	NN	O	O
central	NN	O	O
visual	NN	O	B-Disease
loss	NN	O	I-Disease
developed	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	O
or	NN	O	O
fourth	NN	O	O
decade	NN	O	O
of	NN	O	O
life	NN	O	O
accompanied	NN	O	O
by	NN	O	O
central	NN	O	B-Disease
scotoma	NN	O	I-Disease
and	NN	O	O
well	NN	O	O
-	NN	O	O
demarcated	NN	O	O
atrophy	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
retinal	NN	O	I-Disease
pigment	NN	O	I-Disease
epithelium	NN	O	I-Disease
and	NN	O	O
choriocapillaris	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
macula	NN	O	I-Disease
.	NN	O	O

In	NN	O	O
general	NN	O	O
,	NN	O	O
cone	NN	O	O
and	NN	O	O
rod	NN	O	O
thresholds	NN	O	O
were	NN	O	O
elevated	NN	O	O
,	NN	O	O
and	NN	O	O
color	NN	O	O
-	NN	O	O
contrast	NN	O	O
sensitivity	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
visual	NN	O	O
field	NN	O	O
.	NN	O	O

Peripherally	NN	O	O
,	NN	O	O
the	NN	O	O
scotopic	NN	O	O
sensitivities	NN	O	O
were	NN	O	O
normal	NN	O	O
,	NN	O	O
as	NN	O	O
was	NN	O	O
the	NN	O	O
recovery	NN	O	O
from	NN	O	O
bleach	NN	O	O
.	NN	O	O

Cone	NN	O	O
electroretinograms	NN	O	O
were	NN	O	O
diminished	NN	O	O
in	NN	O	O
amplitude	NN	O	O
,	NN	O	O
and	NN	O	O
delayed	NN	O	O
in	NN	O	O
all	NN	O	O
affected	NN	O	O
adults	NN	O	O
except	NN	O	O
one	NN	O	O
.	NN	O	O

Rod	NN	O	O
electroretinograms	NN	O	O
were	NN	O	O
normal	NN	O	O
or	NN	O	O
near	NN	O	O
normal	NN	O	O
in	NN	O	O
amplitude	NN	O	O
,	NN	O	O
and	NN	O	O
had	NN	O	O
normal	NN	O	O
implicit	NN	O	O
times	NN	O	O
.	NN	O	O

Affected	NN	O	O
asymptomatic	NN	O	O
children	NN	O	O
had	NN	O	O
macular	NN	O	O
changes	NN	O	O
,	NN	O	O
abnormal	NN	O	O
color	NN	O	O
-	NN	O	O
contrast	NN	O	O
sensitivity	NN	O	O
,	NN	O	O
and	NN	O	O
reduced	NN	O	O
pattern	NN	O	O
and	NN	O	O
cone	NN	O	O
electroretinograms	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
rds	NN	O	O
gene	NN	O	O
can	NN	O	O
be	NN	O	O
expressed	NN	O	O
as	NN	O	O
a	NN	O	O
macular	NN	O	B-Disease
dystrophy	NN	O	I-Disease
,	NN	O	O
with	NN	O	O
evidence	NN	O	O
of	NN	O	O
primary	NN	O	O
cone	NN	O	B-Disease
dysfunction	NN	O	I-Disease
and	NN	O	O
preservation	NN	O	O
of	NN	O	O
peripheral	NN	O	O
rod	NN	O	O
function	NN	O	O
.	NN	O	O
.	NN	O	O

Anonymous	NN	O	O
marker	NN	O	O
loci	NN	O	O
within	NN	O	O
400	NN	O	O
kb	NN	O	O
of	NN	O	O
HLA	NN	O	O
-	NN	O	O
A	NN	O	O
generate	NN	O	O
haplotypes	NN	O	O
in	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
with	NN	O	O
the	NN	O	O
hemochromatosis	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
HFE	NN	O	O
)	NN	O	O

The	NN	O	O
hemochromatosis	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
HFE	NN	O	O
)	NN	O	O
maps	NN	O	O
to	NN	O	O
6p21	NN	O	O
.	NN	O	O

3	NN	O	O
and	NN	O	O
is	NN	O	O
less	NN	O	O
than	NN	O	O
1	NN	O	O
cM	NN	O	O
from	NN	O	O
the	NN	O	O
HLA	NN	O	O
class	NN	O	O
I	NN	O	O
genes	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
physical	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
has	NN	O	O
remained	NN	O	O
elusive	NN	O	O
and	NN	O	O
controversial	NN	O	O
.	NN	O	O

The	NN	O	O
unambiguous	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
crossover	NN	O	O
event	NN	O	O
within	NN	O	O
hemochromatosis	NN	O	B-Disease
families	NN	O	O
is	NN	O	O
very	NN	O	O
difficult	NN	O	O
;	NN	O	O
it	NN	O	O
is	NN	O	O
particularly	NN	O	O
hampered	NN	O	O
by	NN	O	O
the	NN	O	O
variability	NN	O	O
of	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
the	NN	O	O
sex	NN	O	O
-	NN	O	O
and	NN	O	O
age	NN	O	O
-	NN	O	O
related	NN	O	O
penetrance	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

For	NN	O	O
these	NN	O	O
practical	NN	O	O
considerations	NN	O	O
,	NN	O	O
traditional	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
could	NN	O	O
prove	NN	O	O
of	NN	O	O
limited	NN	O	O
value	NN	O	O
in	NN	O	O
further	NN	O	O
refining	NN	O	O
the	NN	O	O
extrapolated	NN	O	O
physical	NN	O	O
position	NN	O	O
of	NN	O	O
HFE	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
embarked	NN	O	O
upon	NN	O	O
a	NN	O	O
linkage	NN	O	O
-	NN	O	O
disequilibrium	NN	O	O
analysis	NN	O	O
of	NN	O	O
HFE	NN	O	O
and	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
from	NN	O	O
the	NN	O	O
Brittany	NN	O	O
population	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
66	NN	O	O
hemochromatosis	NN	O	B-Disease
families	NN	O	O
yielding	NN	O	O
151	NN	O	O
hemochromatosis	NN	O	B-Disease
chromosomes	NN	O	O
and	NN	O	O
182	NN	O	O
normal	NN	O	O
chromosomes	NN	O	O
were	NN	O	O
RFLP	NN	O	O
-	NN	O	O
typed	NN	O	O
with	NN	O	O
a	NN	O	O
battery	NN	O	O
of	NN	O	O
probes	NN	O	O
,	NN	O	O
including	NN	O	O
two	NN	O	O
newly	NN	O	O
derived	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
from	NN	O	O
the	NN	O	O
6	NN	O	O
.	NN	O	O

7	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
F	NN	O	O
loci	NN	O	O
located	NN	O	O
150	NN	O	O
and	NN	O	O
250	NN	O	O
kb	NN	O	O
telomeric	NN	O	O
to	NN	O	O
HLA	NN	O	O
-	NN	O	O
A	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
strong	NN	O	O
peak	NN	O	O
of	NN	O	O
existing	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
focused	NN	O	O
within	NN	O	O
the	NN	O	O
i82	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
6	NN	O	O
.	NN	O	O

7	NN	O	O
interval	NN	O	O
(	NN	O	O
approximately	NN	O	O
250	NN	O	O
kb	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
zone	NN	O	O
of	NN	O	O
linkage	NN	O	O
disequilibrium	NN	O	O
is	NN	O	O
flanked	NN	O	O
by	NN	O	O
the	NN	O	O
i97	NN	O	O
locus	NN	O	O
,	NN	O	O
positioned	NN	O	O
30	NN	O	O
kb	NN	O	O
proximal	NN	O	O
to	NN	O	O
i82	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
HLA	NN	O	O
-	NN	O	O
F	NN	O	O
gene	NN	O	O
,	NN	O	O
found	NN	O	O
250	NN	O	O
kb	NN	O	O
distal	NN	O	O
to	NN	O	O
HLA	NN	O	O
-	NN	O	O
A	NN	O	O
,	NN	O	O
markers	NN	O	O
of	NN	O	O
which	NN	O	O
display	NN	O	O
no	NN	O	O
significant	NN	O	O
association	NN	O	O
with	NN	O	O
HFE	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
HFE	NN	O	O
resides	NN	O	O
within	NN	O	O
the	NN	O	O
400	NN	O	O
-	NN	O	O
kb	NN	O	O
expanse	NN	O	O
of	NN	O	O
DNA	NN	O	O
between	NN	O	O
i97	NN	O	O
and	NN	O	O
HLA	NN	O	O
-	NN	O	O
F	NN	O	O
.	NN	O	O

Alternatively	NN	O	O
,	NN	O	O
the	NN	O	O
very	NN	O	O
tight	NN	O	O
association	NN	O	O
of	NN	O	O
HLA	NN	O	O
-	NN	O	O
A3	NN	O	O
and	NN	O	O
allele	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
6	NN	O	O
.	NN	O	O

7	NN	O	O
locus	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
comprised	NN	O	O
by	NN	O	O
the	NN	O	O
major	NN	O	O
ancestral	NN	O	O
or	NN	O	O
founder	NN	O	O
HFE	NN	O	O
haplotype	NN	O	O
in	NN	O	O
Brittany	NN	O	O
,	NN	O	O
supports	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
the	NN	O	O
disease	NN	O	O
gene	NN	O	O
may	NN	O	O
reside	NN	O	O
immediately	NN	O	O
telomeric	NN	O	O
to	NN	O	O
the	NN	O	O
6	NN	O	O
.	NN	O	O

7	NN	O	O
locus	NN	O	O
within	NN	O	O
the	NN	O	O
linkage	NN	O	O
-	NN	O	O
disequilibrium	NN	O	O
zone	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
hemochromatosis	NN	O	B-Disease
haplotypes	NN	O	O
possessing	NN	O	O
HLA	NN	O	O
-	NN	O	O
A11	NN	O	O
and	NN	O	O
the	NN	O	O
low	NN	O	O
-	NN	O	O
frequency	NN	O	O
HLA	NN	O	O
-	NN	O	O
F	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
allele	NN	O	O
2	NN	O	O
)	NN	O	O
are	NN	O	O
supportive	NN	O	O
of	NN	O	O
a	NN	O	O
separate	NN	O	O
founder	NN	O	O
chromosome	NN	O	O
containing	NN	O	O
a	NN	O	O
second	NN	O	O
,	NN	O	O
independently	NN	O	O
arising	NN	O	O
mutant	NN	O	O
allele	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
a	NN	O	O
likely	NN	O	O
"	NN	O	O
hemochromatosis	NN	O	B-Disease
critical	NN	O	O
region	NN	O	O
"	NN	O	O
centromeric	NN	O	O
boundary	NN	O	O
and	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
linkage	NN	O	O
-	NN	O	O
disequilibrium	NN	O	O
zone	NN	O	O
both	NN	O	O
significantly	NN	O	O
contribute	NN	O	O
to	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
DNA	NN	O	O
required	NN	O	O
to	NN	O	O
be	NN	O	O
searched	NN	O	O
for	NN	O	O
novel	NN	O	O
coding	NN	O	O
sequences	NN	O	O
constituting	NN	O	O
the	NN	O	O
HFE	NN	O	B-Disease
defect	NN	O	I-Disease

Genomic	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
EWS	NN	O	O
gene	NN	O	O
and	NN	O	O
its	NN	O	O
relationship	NN	O	O
to	NN	O	O
EWSR1	NN	O	O
,	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
tumor	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
.	NN	O	O

The	NN	O	O
EWS	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
based	NN	O	O
on	NN	O	O
its	NN	O	O
location	NN	O	O
at	NN	O	O
the	NN	O	O
chromosome	NN	O	O
22	NN	O	O
breakpoint	NN	O	O
of	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
11	NN	O	O
;	NN	O	O
22	NN	O	O
)	NN	O	O
(	NN	O	O
q24	NN	O	O
;	NN	O	O
q12	NN	O	O
)	NN	O	O
translocation	NN	O	O
that	NN	O	O
characterizes	NN	O	O
Ewing	NN	O	B-Disease
sarcoma	NN	O	I-Disease
and	NN	O	O
related	NN	O	O
neuroectodermal	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
EWS	NN	O	O
gene	NN	O	O
spans	NN	O	O
about	NN	O	O
40	NN	O	O
kb	NN	O	O
of	NN	O	O
DNA	NN	O	O
and	NN	O	O
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
17	NN	O	O
exons	NN	O	O
.	NN	O	O

The	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
exons	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
cDNA	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
7	NN	O	O
exons	NN	O	O
encode	NN	O	O
the	NN	O	O
N	NN	O	O
-	NN	O	O
terminal	NN	O	O
domain	NN	O	O
of	NN	O	O
EWS	NN	O	O
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
a	NN	O	O
repeated	NN	O	O
degenerated	NN	O	O
polypeptide	NN	O	O
of	NN	O	O
7	NN	O	O
to	NN	O	O
12	NN	O	O
residues	NN	O	O
rich	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
,	NN	O	O
serine	NN	O	O
,	NN	O	O
threonine	NN	O	O
,	NN	O	O
glycine	NN	O	O
,	NN	O	O
and	NN	O	O
glutamine	NN	O	O
.	NN	O	O

Exons	NN	O	O
11	NN	O	O
,	NN	O	O
12	NN	O	O
,	NN	O	O
and	NN	O	O
13	NN	O	O
encode	NN	O	O
the	NN	O	O
putative	NN	O	O
RNA	NN	O	O
binding	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
glycine	NN	O	O
-	NN	O	O
and	NN	O	O
arginine	NN	O	O
-	NN	O	O
rich	NN	O	O
motifs	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
are	NN	O	O
mainly	NN	O	O
encoded	NN	O	O
by	NN	O	O
exons	NN	O	O
8	NN	O	O
-	NN	O	O
9	NN	O	O
,	NN	O	O
14	NN	O	O
,	NN	O	O
and	NN	O	O
16	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
has	NN	O	O
features	NN	O	O
of	NN	O	O
a	NN	O	O
CpG	NN	O	O
-	NN	O	O
rich	NN	O	O
island	NN	O	O
and	NN	O	O
lacks	NN	O	O
canonical	NN	O	O
promoter	NN	O	O
elements	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
TATA	NN	O	O
and	NN	O	O
CCAAT	NN	O	O
consensus	NN	O	O
sequences	NN	O	O
.	NN	O	O

Positions	NN	O	O
of	NN	O	O
the	NN	O	O
chromosome	NN	O	O
22	NN	O	O
breakpoints	NN	O	O
were	NN	O	O
determined	NN	O	O
for	NN	O	O
19	NN	O	O
Ewing	NN	O	B-Disease
tumors	NN	O	I-Disease
.	NN	O	O

They	NN	O	O
were	NN	O	O
localized	NN	O	O
in	NN	O	O
introns	NN	O	O
7	NN	O	O
or	NN	O	O
8	NN	O	O
in	NN	O	O
18	NN	O	O
cases	NN	O	O
and	NN	O	O
in	NN	O	O
intron	NN	O	O
10	NN	O	O
in	NN	O	O
1	NN	O	O
case	NN	O	O
.	NN	O	O
.	NN	O	O

Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
gene	NN	O	O
:	NN	O	O
characterization	NN	O	O
of	NN	O	O
deletions	NN	O	O
and	NN	O	O
possible	NN	O	O
function	NN	O	O
.	NN	O	O

Positional	NN	O	O
cloning	NN	O	O
experiments	NN	O	O
have	NN	O	O
resulted	NN	O	O
recently	NN	O	O
in	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
pseudoglioma	NN	O	B-Disease
;	NN	O	O
NDP	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
severe	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
neurodevelopmental	NN	O	I-Disease
disorder	NN	O	I-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
clones	NN	O	O
encompassing	NN	O	O
the	NN	O	O
NDP	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
spans	NN	O	O
28	NN	O	O
kb	NN	O	O
and	NN	O	O
consists	NN	O	O
of	NN	O	O
3	NN	O	O
exons	NN	O	O
,	NN	O	O
the	NN	O	O
first	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
entirely	NN	O	O
contained	NN	O	O
within	NN	O	O
the	NN	O	O
5	NN	O	O
untranslated	NN	O	O
region	NN	O	O
.	NN	O	O

Detailed	NN	O	O
analysis	NN	O	O
of	NN	O	O
genomic	NN	O	O
deletions	NN	O	O
in	NN	O	O
Norrie	NN	O	B-Disease
patients	NN	O	O
shows	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
heterogeneous	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
size	NN	O	O
and	NN	O	O
in	NN	O	O
position	NN	O	O
.	NN	O	O

By	NN	O	O
PCR	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NDP	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
not	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
eye	NN	O	O
or	NN	O	O
to	NN	O	O
the	NN	O	O
brain	NN	O	O
.	NN	O	O

An	NN	O	O
extensive	NN	O	O
DNA	NN	O	O
and	NN	O	O
protein	NN	O	O
sequence	NN	O	O
comparison	NN	O	O
between	NN	O	O
the	NN	O	O
human	NN	O	O
NDP	NN	O	B-Disease
gene	NN	O	O
and	NN	O	O
related	NN	O	O
genes	NN	O	O
from	NN	O	O
the	NN	O	O
database	NN	O	O
revealed	NN	O	O
homology	NN	O	O
with	NN	O	O
cysteine	NN	O	O
-	NN	O	O
rich	NN	O	O
protein	NN	O	O
-	NN	O	O
binding	NN	O	O
domains	NN	O	O
of	NN	O	O
immediate	NN	O	O
-	NN	O	O
-	NN	O	O
early	NN	O	O
genes	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
NDP	NN	O	O
is	NN	O	O
a	NN	O	O
molecule	NN	O	O
related	NN	O	O
in	NN	O	O
function	NN	O	O
to	NN	O	O
these	NN	O	O
genes	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
a	NN	O	O
pathway	NN	O	O
that	NN	O	O
regulates	NN	O	O
neural	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
normal	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
allele	NN	O	O
,	NN	O	O
or	NN	O	O
a	NN	O	O
closely	NN	O	O
linked	NN	O	O
gene	NN	O	O
,	NN	O	O
influences	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
of	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
(	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
normal	NN	O	O
HD	NN	O	B-Disease
allele	NN	O	O
by	NN	O	O
comparing	NN	O	O
transmission	NN	O	O
patterns	NN	O	O
of	NN	O	O
genetically	NN	O	O
linked	NN	O	O
markers	NN	O	O
at	NN	O	O
the	NN	O	O
D4S10	NN	O	O
locus	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
parent	NN	O	O
against	NN	O	O
age	NN	O	O
at	NN	O	O
onset	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
offspring	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
information	NN	O	O
from	NN	O	O
21	NN	O	O
sibships	NN	O	O
in	NN	O	O
14	NN	O	O
kindreds	NN	O	O
showed	NN	O	O
a	NN	O	O
significant	NN	O	O
tendency	NN	O	O
for	NN	O	O
sibs	NN	O	O
who	NN	O	O
have	NN	O	O
similar	NN	O	O
onset	NN	O	O
ages	NN	O	O
to	NN	O	O
share	NN	O	O
the	NN	O	O
same	NN	O	O
D4S10	NN	O	O
allele	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
parent	NN	O	O
.	NN	O	O

Affected	NN	O	O
sibs	NN	O	O
who	NN	O	O
inherited	NN	O	O
different	NN	O	O
D4S10	NN	O	O
alleles	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
parent	NN	O	O
tended	NN	O	O
to	NN	O	O
have	NN	O	O
more	NN	O	O
variable	NN	O	O
ages	NN	O	O
at	NN	O	O
onset	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
HD	NN	O	B-Disease
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
the	NN	O	O
normal	NN	O	O
HD	NN	O	B-Disease
allele	NN	O	O
or	NN	O	O
by	NN	O	O
a	NN	O	O
closely	NN	O	O
linked	NN	O	O
locus	NN	O	O
.	NN	O	O
.	NN	O	O

Further	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
intron	NN	O	O
9	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
mutation	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
patients	NN	O	O
from	NN	O	O
the	NN	O	O
British	NN	O	O
Isles	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
previous	NN	O	O
study	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
Tay	NN	O	B-Disease
-	NN	O	I-Disease
Sachs	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
TSD	NN	O	B-Disease
)	NN	O	O
causing	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
intron	NN	O	O
9	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
HEXA	NN	O	O
gene	NN	O	O
occurs	NN	O	O
at	NN	O	O
high	NN	O	O
frequency	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
patients	NN	O	O
and	NN	O	O
carriers	NN	O	O
from	NN	O	O
the	NN	O	O
British	NN	O	O
Isles	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
more	NN	O	O
frequently	NN	O	O
in	NN	O	O
subjects	NN	O	O
of	NN	O	O
Irish	NN	O	O
,	NN	O	O
Scottish	NN	O	O
,	NN	O	O
and	NN	O	O
Welsh	NN	O	O
origin	NN	O	O
compared	NN	O	O
with	NN	O	O
English	NN	O	O
origin	NN	O	O
(	NN	O	O
63	NN	O	O
%	NN	O	O
and	NN	O	O
31	NN	O	O
%	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
tested	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
blind	NN	O	O
study	NN	O	O
,	NN	O	O
26	NN	O	O
American	NN	O	O
TSD	NN	O	B-Disease
carriers	NN	O	O
and	NN	O	O
28	NN	O	O
non	NN	O	O
-	NN	O	O
carriers	NN	O	O
who	NN	O	O
have	NN	O	O
British	NN	O	O
ancestry	NN	O	O
for	NN	O	O
the	NN	O	O
intron	NN	O	O
9	NN	O	O
splice	NN	O	O
site	NN	O	O
mutation	NN	O	O
.	NN	O	O

Six	NN	O	O
of	NN	O	O
the	NN	O	O
carriers	NN	O	O
and	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
controls	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

All	NN	O	O
six	NN	O	O
had	NN	O	O
Irish	NN	O	O
ancestry	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
nine	NN	O	O
of	NN	O	O
the	NN	O	O
20	NN	O	O
other	NN	O	O
(	NN	O	O
intron	NN	O	O
9	NN	O	O
mutation	NN	O	O
negative	NN	O	O
)	NN	O	O
TSD	NN	O	B-Disease
carriers	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
confirm	NN	O	O
the	NN	O	O
previously	NN	O	O
found	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	O
9	NN	O	O
mutation	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Jewish	NN	O	O
TSD	NN	O	B-Disease
families	NN	O	O
of	NN	O	O
British	NN	O	O
Isles	NN	O	O
,	NN	O	O
particularly	NN	O	O
Irish	NN	O	O
,	NN	O	O
origin	NN	O	O
,	NN	O	O
and	NN	O	O
reinforce	NN	O	O
the	NN	O	O
need	NN	O	O
to	NN	O	O
screen	NN	O	O
such	NN	O	O
families	NN	O	O
for	NN	O	O
this	NN	O	O
mutation	NN	O	O
.	NN	O	O

Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
oncogenic	NN	O	O
mutations	NN	O	O
in	NN	O	O
tumors	NN	O	B-Disease
from	NN	O	O
patients	NN	O	O
with	NN	O	O
bilateral	NN	O	B-Disease
and	NN	O	I-Disease
unilateral	NN	O	I-Disease
retinoblastoma	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
RB1	NN	O	O
gene	NN	O	O
from	NN	O	O
12	NN	O	O
human	NN	O	O
retinoblastoma	NN	O	B-Disease
tumors	NN	O	I-Disease
has	NN	O	O
been	NN	O	O
analyzed	NN	O	O
exon	NN	O	O
-	NN	O	O
by	NN	O	O
-	NN	O	O
exon	NN	O	O
with	NN	O	O
the	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
technique	NN	O	O
.	NN	O	O

Mutations	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
tumors	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
one	NN	O	O
-	NN	O	O
third	NN	O	O
of	NN	O	O
the	NN	O	O
tumors	NN	O	B-Disease
had	NN	O	O
independent	NN	O	O
mutations	NN	O	O
in	NN	O	O
both	NN	O	O
alleles	NN	O	O
neither	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
germ	NN	O	O
line	NN	O	O
,	NN	O	O
confirming	NN	O	O
their	NN	O	O
true	NN	O	O
sporadic	NN	O	O
nature	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
remaining	NN	O	O
two	NN	O	O
-	NN	O	O
thirds	NN	O	O
of	NN	O	O
the	NN	O	O
tumors	NN	O	B-Disease
only	NN	O	O
one	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
heterozygosity	NN	O	O
theory	NN	O	O
of	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
,	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transitions	NN	O	O
,	NN	O	O
were	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
abnormality	NN	O	O
and	NN	O	O
usually	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
an	NN	O	O
arginine	NN	O	O
codon	NN	O	O
to	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
.	NN	O	O

Small	NN	O	O
deletions	NN	O	O
were	NN	O	O
the	NN	O	O
second	NN	O	O
most	NN	O	O
common	NN	O	O
abnormality	NN	O	O
and	NN	O	O
most	NN	O	O
often	NN	O	O
created	NN	O	O
a	NN	O	O
downstream	NN	O	O
stop	NN	O	O
codon	NN	O	O
as	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
reading	NN	O	O
frameshift	NN	O	O
.	NN	O	O

Deletions	NN	O	O
and	NN	O	O
point	NN	O	O
mutations	NN	O	O
also	NN	O	O
affected	NN	O	O
splice	NN	O	O
junctions	NN	O	O
.	NN	O	O

Direct	NN	O	O
repeats	NN	O	O
were	NN	O	O
present	NN	O	O
at	NN	O	O
the	NN	O	O
breakpoint	NN	O	O
junctions	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
deletions	NN	O	O
,	NN	O	O
supporting	NN	O	O
a	NN	O	O
slipped	NN	O	O
-	NN	O	O
mispairing	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
generally	NN	O	O
produced	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
which	NN	O	O
resulted	NN	O	O
in	NN	O	O
perfect	NN	O	O
homology	NN	O	O
with	NN	O	O
endogenous	NN	O	O
sequences	NN	O	O
which	NN	O	O
lay	NN	O	O
within	NN	O	O
14	NN	O	O
bp	NN	O	O
.	NN	O	O
.	NN	O	O

PAX6	NN	O	O
mutations	NN	O	O
in	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O

Aniridia	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
congenital	NN	O	B-Disease
malformation	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
eye	NN	O	I-Disease
,	NN	O	O
chiefly	NN	O	O
characterised	NN	O	O
by	NN	O	O
iris	NN	O	B-Disease
hypoplasia	NN	O	I-Disease
,	NN	O	O
which	NN	O	O
can	NN	O	O
cause	NN	O	O
blindness	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
PAX6	NN	O	O
gene	NN	O	O
was	NN	O	O
isolated	NN	O	O
as	NN	O	O
a	NN	O	O
candidate	NN	O	O
aniridia	NN	O	B-Disease
gene	NN	O	O
by	NN	O	O
positional	NN	O	O
cloning	NN	O	O
from	NN	O	O
the	NN	O	O
smallest	NN	O	O
region	NN	O	O
of	NN	O	O
overlap	NN	O	O
of	NN	O	O
aniridia	NN	O	B-Disease
-	NN	O	O
associated	NN	O	O
deletions	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
PAX6	NN	O	O
intragenic	NN	O	O
mutations	NN	O	O
were	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
Smalleye	NN	O	O
,	NN	O	O
a	NN	O	O
mouse	NN	O	O
mutant	NN	O	O
which	NN	O	O
is	NN	O	O
an	NN	O	O
animal	NN	O	O
model	NN	O	O
for	NN	O	O
aniridia	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
six	NN	O	O
human	NN	O	O
aniridia	NN	O	B-Disease
patients	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
we	NN	O	O
describe	NN	O	O
four	NN	O	O
additional	NN	O	O
PAX6	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
aniridia	NN	O	B-Disease
patients	NN	O	O
,	NN	O	O
both	NN	O	O
sporadic	NN	O	O
and	NN	O	O
familial	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
highlight	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
which	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
PAX6	NN	O	O
function	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
frequency	NN	O	O
at	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
PAX6	NN	O	O
mutations	NN	O	O
suggests	NN	O	O
that	NN	O	O
lesions	NN	O	O
in	NN	O	O
PAX6	NN	O	O
will	NN	O	O
account	NN	O	O
for	NN	O	O
most	NN	O	O
cases	NN	O	O
of	NN	O	O
aniridia	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
arginine	NN	O	O
vasopressin	NN	O	O
defect	NN	O	O
by	NN	O	O
dideoxy	NN	O	O
fingerprinting	NN	O	O
.	NN	O	O

Autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
is	NN	O	O
a	NN	O	O
familial	NN	O	O
form	NN	O	O
of	NN	O	O
diabetes	NN	O	B-Disease
insipidus	NN	O	I-Disease
.	NN	O	O

This	NN	O	O
disorder	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
variable	NN	O	O
levels	NN	O	O
of	NN	O	O
arginine	NN	O	O
vasopressin	NN	O	O
(	NN	O	O
AVP	NN	O	O
)	NN	O	O
and	NN	O	O
diabetes	NN	O	B-Disease
insipidus	NN	O	I-Disease
of	NN	O	O
varying	NN	O	O
severity	NN	O	O
,	NN	O	O
which	NN	O	O
responds	NN	O	O
to	NN	O	O
exogenous	NN	O	O
AVP	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
autosomal	NN	O	B-Disease
dominant	NN	O	I-Disease
neurohypophyseal	NN	O	I-Disease
diabetes	NN	O	I-Disease
insipidus	NN	O	I-Disease
,	NN	O	O
the	NN	O	O
AVP	NN	O	O
genes	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
kindred	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
method	NN	O	O
,	NN	O	O
called	NN	O	O
dideoxy	NN	O	O
fingerprinting	NN	O	O
,	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
an	NN	O	O
AVP	NN	O	O
mutation	NN	O	O
that	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
.	NN	O	O

The	NN	O	O
novel	NN	O	O
defect	NN	O	O
found	NN	O	O
changes	NN	O	O
the	NN	O	O
last	NN	O	O
codon	NN	O	O
of	NN	O	O
the	NN	O	O
AVP	NN	O	O
signal	NN	O	O
peptide	NN	O	O
from	NN	O	O
alanine	NN	O	O
to	NN	O	O
threonine	NN	O	O
,	NN	O	O
which	NN	O	O
should	NN	O	O
perturb	NN	O	O
cleavage	NN	O	O
of	NN	O	O
mature	NN	O	O
AVP	NN	O	O
from	NN	O	O
its	NN	O	O
precursor	NN	O	O
protein	NN	O	O
and	NN	O	O
inhibit	NN	O	O
its	NN	O	O
secretion	NN	O	O
or	NN	O	O
action	NN	O	O
.	NN	O	O
.	NN	O	O

Germinal	NN	O	O
mosaicism	NN	O	O
in	NN	O	O
a	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
family	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
genetic	NN	O	O
counselling	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
describe	NN	O	O
a	NN	O	O
three	NN	O	O
-	NN	O	O
generation	NN	O	O
family	NN	O	O
in	NN	O	O
which	NN	O	O
two	NN	O	O
siblings	NN	O	O
were	NN	O	O
affected	NN	O	O
by	NN	O	O
Duchenne	NN	O	B-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
analysis	NN	O	O
of	NN	O	O
muscle	NN	O	O
dystrophin	NN	O	O
and	NN	O	O
haplotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
locus	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
carrying	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
was	NN	O	O
transmitted	NN	O	O
from	NN	O	O
the	NN	O	O
healthy	NN	O	O
maternal	NN	O	O
grandfather	NN	O	O
to	NN	O	O
his	NN	O	O
three	NN	O	O
daughters	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
probands	NN	O	O
mother	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
grandfather	NN	O	O
was	NN	O	O
a	NN	O	O
germinal	NN	O	O
mosaic	NN	O	O
for	NN	O	O
the	NN	O	O
DMD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

The	NN	O	O
definition	NN	O	O
of	NN	O	O
the	NN	O	O
carrier	NN	O	O
status	NN	O	O
in	NN	O	O
two	NN	O	O
possible	NN	O	O
carriers	NN	O	O
led	NN	O	O
us	NN	O	O
to	NN	O	O
give	NN	O	O
accurate	NN	O	O
genetic	NN	O	O
counselling	NN	O	O
and	NN	O	O
to	NN	O	O
prevent	NN	O	O
the	NN	O	O
birth	NN	O	O
of	NN	O	O
an	NN	O	O
affected	NN	O	O
boy	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
usefulness	NN	O	O
of	NN	O	O
haplotype	NN	O	O
analysis	NN	O	O
and	NN	O	O
immunohistochemical	NN	O	O
muscle	NN	O	O
dystrophin	NN	O	O
studies	NN	O	O
to	NN	O	O
detect	NN	O	O
hidden	NN	O	O
germinal	NN	O	O
mosaicism	NN	O	O
and	NN	O	O
to	NN	O	O
improve	NN	O	O
genetic	NN	O	O
counselling	NN	O	O
.	NN	O	O
.	NN	O	O

Genetic	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
to	NN	O	O
a	NN	O	O
small	NN	O	O
interval	NN	O	O
on	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
-	NN	O	O
21	NN	O	O
:	NN	O	O
exclusion	NN	O	O
of	NN	O	O
candidate	NN	O	O
genes	NN	O	O
EDH17B2	NN	O	O
and	NN	O	O
RARA	NN	O	O
.	NN	O	O

A	NN	O	O
susceptibility	NN	O	O
gene	NN	O	O
for	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
assigned	NN	O	O
by	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
to	NN	O	O
chromosome	NN	O	O
17q21	NN	O	O
.	NN	O	O

Candidate	NN	O	O
genes	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
include	NN	O	O
EDH17B2	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
estradiol	NN	O	O
17	NN	O	O
beta	NN	O	O
-	NN	O	O
hydroxysteroid	NN	O	O
dehydrogenase	NN	O	O
II	NN	O	O
(	NN	O	O
17	NN	O	O
beta	NN	O	O
-	NN	O	O
HSD	NN	O	O
II	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
RARA	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
receptor	NN	O	O
alpha	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
typed	NN	O	O
22	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
breast	NN	O	I-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
with	NN	O	O
eight	NN	O	O
polymorphisms	NN	O	O
from	NN	O	O
the	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
-	NN	O	O
21	NN	O	O
region	NN	O	O
,	NN	O	O
including	NN	O	O
two	NN	O	O
in	NN	O	O
the	NN	O	O
EDH17B2	NN	O	O
gene	NN	O	O
.	NN	O	O

Genetic	NN	O	O
recombination	NN	O	O
with	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
trait	NN	O	O
excludes	NN	O	O
RARA	NN	O	O
from	NN	O	O
further	NN	O	O
consideration	NN	O	O
as	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Both	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
EDH17B2	NN	O	O
map	NN	O	O
to	NN	O	O
a	NN	O	O
6	NN	O	O
cM	NN	O	O
interval	NN	O	O
(	NN	O	O
between	NN	O	O
THRA1	NN	O	O
and	NN	O	O
D17S579	NN	O	O
)	NN	O	O
and	NN	O	O
no	NN	O	O
recombination	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
genes	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
overlapping	NN	O	O
PCR	NN	O	O
products	NN	O	O
containing	NN	O	O
the	NN	O	O
entire	NN	O	O
EDH17B2	NN	O	O
gene	NN	O	O
in	NN	O	O
four	NN	O	O
unrelated	NN	O	O
affected	NN	O	O
women	NN	O	O
did	NN	O	O
not	NN	O	O
uncover	NN	O	O
any	NN	O	O
sequence	NN	O	O
variation	NN	O	O
,	NN	O	O
other	NN	O	O
than	NN	O	O
previously	NN	O	O
described	NN	O	O
polymorphisms	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
EDH17B2	NN	O	O
gene	NN	O	O
,	NN	O	O
therefore	NN	O	O
do	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
hereditary	NN	O	B-Disease
breast	NN	O	I-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

Single	NN	O	O
meiotic	NN	O	O
crossovers	NN	O	O
in	NN	O	O
affected	NN	O	O
women	NN	O	O
suggest	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
is	NN	O	O
flanked	NN	O	O
by	NN	O	O
the	NN	O	O
loci	NN	O	O
RARA	NN	O	O
and	NN	O	O
D17S78	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
cholesteryl	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
gene	NN	O	O
with	NN	O	O
possible	NN	O	O
dominant	NN	O	O
effects	NN	O	O
on	NN	O	O
plasma	NN	O	O
high	NN	O	O
density	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

Plasma	NN	O	O
HDL	NN	O	O
are	NN	O	O
a	NN	O	O
negative	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
atherosclerosis	NN	O	B-Disease
.	NN	O	O

Cholesteryl	NN	O	O
ester	NN	O	O
transfer	NN	O	O
protein	NN	O	O
(	NN	O	O
CETP	NN	O	O
;	NN	O	O
476	NN	O	O
amino	NN	O	O
acids	NN	O	O
)	NN	O	O
transfers	NN	O	O
cholesteryl	NN	O	O
ester	NN	O	O
from	NN	O	O
HDL	NN	O	O
to	NN	O	O
other	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

Subjects	NN	O	O
with	NN	O	O
homozygous	NN	O	O
CETP	NN	O	B-Disease
deficiency	NN	O	I-Disease
caused	NN	O	O
by	NN	O	O
a	NN	O	O
gene	NN	O	O
splicing	NN	O	O
defect	NN	O	O
have	NN	O	O
markedly	NN	O	O
elevated	NN	O	O
HDL	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
heterozygotes	NN	O	O
have	NN	O	O
only	NN	O	O
mild	NN	O	O
increases	NN	O	O
in	NN	O	O
HDL	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
two	NN	O	O
probands	NN	O	O
with	NN	O	O
a	NN	O	O
CETP	NN	O	O
missense	NN	O	O
mutation	NN	O	O
(	NN	O	O
442	NN	O	O
D	NN	O	O
G	NN	O	O
)	NN	O	O
.	NN	O	O

Although	NN	O	O
heterozygous	NN	O	O
,	NN	O	O
they	NN	O	O
have	NN	O	O
threefold	NN	O	O
increases	NN	O	O
in	NN	O	O
HDL	NN	O	O
concentration	NN	O	O
and	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
CETP	NN	O	O
mass	NN	O	O
and	NN	O	O
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
has	NN	O	O
dominant	NN	O	O
effects	NN	O	O
on	NN	O	O
CETP	NN	O	O
and	NN	O	O
HDL	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Cellular	NN	O	O
expression	NN	O	O
of	NN	O	O
mutant	NN	O	O
cDNA	NN	O	O
results	NN	O	O
in	NN	O	O
secretion	NN	O	O
of	NN	O	O
only	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
CETP	NN	O	O
activity	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
and	NN	O	O
mutant	NN	O	O
cDNAs	NN	O	O
leads	NN	O	O
to	NN	O	O
inhibition	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
secretion	NN	O	O
and	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
dominant	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
CETP	NN	O	O
missense	NN	O	O
mutation	NN	O	O
during	NN	O	O
cellular	NN	O	O
expression	NN	O	O
probably	NN	O	O
explains	NN	O	O
why	NN	O	O
the	NN	O	O
probands	NN	O	O
have	NN	O	O
markedly	NN	O	O
increased	NN	O	O
HDL	NN	O	O
in	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
state	NN	O	O
,	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
active	NN	O	O
molecular	NN	O	O
species	NN	O	O
of	NN	O	O
CETP	NN	O	O
may	NN	O	O
be	NN	O	O
multimeric	NN	O	O
.	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
colchicine	NN	O	O
era	NN	O	O
:	NN	O	O
the	NN	O	O
fate	NN	O	O
of	NN	O	O
one	NN	O	O
family	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
prophylactic	NN	O	O
colchicine	NN	O	O
treatment	NN	O	O
on	NN	O	O
the	NN	O	O
natural	NN	O	O
history	NN	O	O
of	NN	O	O
familial	NN	O	B-Disease
Mediterranean	NN	O	I-Disease
fever	NN	O	I-Disease
(	NN	O	O
FMF	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
family	NN	O	O
is	NN	O	O
presented	NN	O	O
with	NN	O	O
6	NN	O	O
out	NN	O	O
of	NN	O	O
9	NN	O	O
siblings	NN	O	O
affected	NN	O	O
by	NN	O	O
FMF	NN	O	B-Disease
.	NN	O	O

Each	NN	O	O
patient	NN	O	O
represents	NN	O	O
a	NN	O	O
different	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
amyloidotic	NN	O	B-Disease
kidney	NN	O	I-Disease
disease	NN	O	I-Disease
of	NN	O	O
FMF	NN	O	B-Disease
and	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
continuous	NN	O	O
colchicine	NN	O	O
treatment	NN	O	O
on	NN	O	O
its	NN	O	O
course	NN	O	O
.	NN	O	O

Considered	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
members	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
present	NN	O	O
an	NN	O	O
almost	NN	O	O
complete	NN	O	O
clinical	NN	O	O
,	NN	O	O
genetic	NN	O	O
,	NN	O	O
and	NN	O	O
behavioral	NN	O	O
picture	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
submicroscopic	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
deletion	NN	O	O
interval	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
DNA	NN	O	O
probe	NN	O	O
isolated	NN	O	O
by	NN	O	O
differential	NN	O	O
Alu	NN	O	O
PCR	NN	O	O
fingerprint	NN	O	O
cloning	NN	O	O
.	NN	O	O

Differential	NN	O	O
Alu	NN	O	O
PCR	NN	O	O
fingerprint	NN	O	O
cloning	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
a	NN	O	O
DNA	NN	O	O
probe	NN	O	O
from	NN	O	O
the	NN	O	O
Xp11	NN	O	O
.	NN	O	O

4	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
p11	NN	O	O
.	NN	O	O

21	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
X	NN	O	O
chromosome	NN	O	O
.	NN	O	O

This	NN	O	O
novel	NN	O	O
sequence	NN	O	O
,	NN	O	O
cpXr318	NN	O	O
(	NN	O	O
DXS742	NN	O	O
)	NN	O	O
,	NN	O	O
detects	NN	O	O
a	NN	O	O
new	NN	O	O
submicroscopic	NN	O	O
deletion	NN	O	O
interval	NN	O	O
at	NN	O	O
the	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
locus	NN	O	O
(	NN	O	O
NDP	NN	O	O
)	NN	O	O
.	NN	O	O

Combining	NN	O	O
our	NN	O	O
data	NN	O	O
with	NN	O	O
the	NN	O	O
consensus	NN	O	O
genetic	NN	O	O
map	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	O
short	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	O
chromosome	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
the	NN	O	O
physical	NN	O	O
order	NN	O	O
Xcen	NN	O	O
-	NN	O	O
DXS14	NN	O	O
-	NN	O	O
DXS255	NN	O	O
-	NN	O	O
(	NN	O	O
DXS426	NN	O	O
,	NN	O	O
TIMP	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
DXS742	NN	O	O
-	NN	O	O
(	NN	O	O
[	NN	O	O
MAOB	NN	O	O
-	NN	O	O
MAOA	NN	O	O
-	NN	O	O
DXS7	NN	O	O
]	NN	O	O
,	NN	O	O
NDP	NN	O	O
)	NN	O	O
-	NN	O	O
DXS77	NN	O	O
-	NN	O	O
DXS228	NN	O	O
)	NN	O	O
-	NN	O	O
DXS209	NN	O	O
-	NN	O	O
DXS148	NN	O	O
-	NN	O	O
DXS196	NN	O	O
-	NN	O	O
+	NN	O	O
+	NN	O	O
+	NN	O	O
Xpter	NN	O	O
.	NN	O	O

The	NN	O	O
cpXr318	NN	O	O
probe	NN	O	O
and	NN	O	O
a	NN	O	O
subclone	NN	O	O
from	NN	O	O
a	NN	O	O
cosmid	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
DXS7	NN	O	O
locus	NN	O	O
were	NN	O	O
converted	NN	O	O
into	NN	O	O
sequence	NN	O	O
-	NN	O	O
tagged	NN	O	O
sites	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
DXS742	NN	O	O
,	NN	O	O
DSX7	NN	O	O
,	NN	O	O
DXS77	NN	O	O
,	NN	O	O
and	NN	O	O
MAOA	NN	O	O
were	NN	O	O
integrated	NN	O	O
into	NN	O	O
a	NN	O	O
physical	NN	O	O
map	NN	O	O
spanning	NN	O	O
the	NN	O	O
Norrie	NN	O	B-Disease
disease	NN	O	I-Disease
locus	NN	O	O

Putative	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
adrenoleukodystrophy	NN	O	I-Disease
gene	NN	O	O
shares	NN	O	O
unexpected	NN	O	O
homology	NN	O	O
with	NN	O	O
ABC	NN	O	O
transporters	NN	O	O
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
is	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
disease	NN	O	I-Disease
affecting	NN	O	O
1	NN	O	O
/	NN	O	O
20	NN	O	O
,	NN	O	O
000	NN	O	O
males	NN	O	O
either	NN	O	O
as	NN	O	O
cerebral	NN	O	B-Disease
ALD	NN	O	I-Disease
in	NN	O	O
childhood	NN	O	O
or	NN	O	O
as	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
adults	NN	O	O
.	NN	O	O

Childhood	NN	O	O
ALD	NN	O	B-Disease
is	NN	O	O
the	NN	O	O
more	NN	O	O
severe	NN	O	O
form	NN	O	O
,	NN	O	O
with	NN	O	O
onset	NN	O	O
of	NN	O	O
neurological	NN	O	O
symptoms	NN	O	O
between	NN	O	O
5	NN	O	O
-	NN	O	O
12	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

Central	NN	O	B-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
demyelination	NN	O	I-Disease
progresses	NN	O	O
rapidly	NN	O	O
and	NN	O	O
death	NN	O	O
occurs	NN	O	O
within	NN	O	O
a	NN	O	O
few	NN	O	O
years	NN	O	O
.	NN	O	O

AMN	NN	O	B-Disease
is	NN	O	O
a	NN	O	O
milder	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
with	NN	O	O
onset	NN	O	O
at	NN	O	O
15	NN	O	O
-	NN	O	O
30	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
and	NN	O	O
a	NN	O	O
more	NN	O	O
progressive	NN	O	O
course	NN	O	O
.	NN	O	O

Adrenal	NN	O	B-Disease
insufficiency	NN	O	I-Disease
(	NN	O	O
Addisons	NN	O	B-Disease
disease	NN	O	I-Disease
)	NN	O	O
may	NN	O	O
remain	NN	O	O
the	NN	O	O
only	NN	O	O
clinical	NN	O	O
manifestation	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
principal	NN	O	O
biochemical	NN	O	O
abnormality	NN	O	O
of	NN	O	O
ALD	NN	O	B-Disease
is	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
(	NN	O	O
VLCFA	NN	O	O
)	NN	O	O
because	NN	O	O
of	NN	O	O
impaired	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
in	NN	O	O
peroxisomes	NN	O	O
.	NN	O	O

The	NN	O	O
normal	NN	O	O
oxidation	NN	O	O
of	NN	O	O
VLCFA	NN	O	O
-	NN	O	O
CoA	NN	O	O
in	NN	O	O
patients	NN	O	O
fibroblasts	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
VLCFA	NN	O	O
-	NN	O	O
CoA	NN	O	O
synthetase	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
for	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
use	NN	O	O
positional	NN	O	O
cloning	NN	O	O
to	NN	O	O
identify	NN	O	O
a	NN	O	O
gene	NN	O	O
partially	NN	O	O
deleted	NN	O	O
in	NN	O	O
6	NN	O	O
of	NN	O	O
85	NN	O	O
independent	NN	O	O
patients	NN	O	O
with	NN	O	O
ALD	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
familial	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
deletions	NN	O	O
segregated	NN	O	O
with	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

An	NN	O	O
identical	NN	O	O
deletion	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
two	NN	O	O
brothers	NN	O	O
presenting	NN	O	O
with	NN	O	O
different	NN	O	O
clinical	NN	O	O
ALD	NN	O	B-Disease
phenotypes	NN	O	O
.	NN	O	O

Candidate	NN	O	O
exons	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
computer	NN	O	O
analysis	NN	O	O
of	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
and	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
complementary	NN	O	O
DNAs	NN	O	O
by	NN	O	O
exon	NN	O	O
connection	NN	O	O
and	NN	O	O
screening	NN	O	O
of	NN	O	O
cDNA	NN	O	O
libraries	NN	O	O
.	NN	O	O

The	NN	O	O
deduced	NN	O	O
protein	NN	O	O
sequence	NN	O	O
shows	NN	O	O
significant	NN	O	O
sequence	NN	O	O
identity	NN	O	O
to	NN	O	O
a	NN	O	O
peroxisomal	NN	O	O
membrane	NN	O	O
protein	NN	O	O
of	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
70K	NN	O	O
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
peroxisome	NN	O	O
biogenesis	NN	O	O
and	NN	O	O
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
ATP	NN	O	O
-	NN	O	O
binding	NN	O	O
cassette	NN	O	O
superfamily	NN	O	O
of	NN	O	O
transporters	NN	O	O
.	NN	O	O
.	NN	O	O

Targeted	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
apolipoprotein	NN	O	O
B	NN	O	O
gene	NN	O	O
results	NN	O	O
in	NN	O	O
hypobetalipoproteinemia	NN	O	B-Disease
and	NN	O	O
developmental	NN	O	B-Disease
abnormalities	NN	O	I-Disease
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Familial	NN	O	B-Disease
hypobetalipoproteinemia	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
autosomal	NN	O	B-Disease
codominant	NN	O	I-Disease
disorder	NN	O	I-Disease
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
dramatic	NN	O	O
reduction	NN	O	O
in	NN	O	O
plasma	NN	O	O
concentrations	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
(	NN	O	O
apo	NN	O	O
)	NN	O	O
B	NN	O	O
,	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
and	NN	O	O
beta	NN	O	O
-	NN	O	O
migrating	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

A	NN	O	O
benefit	NN	O	O
of	NN	O	O
hypobetalipoproteinemia	NN	O	B-Disease
is	NN	O	O
that	NN	O	O
mildly	NN	O	O
affected	NN	O	O
individuals	NN	O	O
may	NN	O	O
be	NN	O	O
protected	NN	O	O
from	NN	O	O
coronary	NN	O	B-Disease
vascular	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
gene	NN	O	O
targeting	NN	O	O
to	NN	O	O
generate	NN	O	O
mice	NN	O	O
with	NN	O	O
a	NN	O	O
modified	NN	O	O
Apob	NN	O	O
allele	NN	O	O
.	NN	O	O

Mice	NN	O	O
containing	NN	O	O
this	NN	O	O
allele	NN	O	O
display	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
hallmarks	NN	O	O
of	NN	O	O
human	NN	O	O
hypobetalipoproteinemia	NN	O	B-Disease
they	NN	O	O
produce	NN	O	O
a	NN	O	O
truncated	NN	O	O
apoB	NN	O	O
protein	NN	O	O
,	NN	O	O
apoB70	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
concentrations	NN	O	O
of	NN	O	O
apoB	NN	O	O
,	NN	O	O
beta	NN	O	O
-	NN	O	O
lipoproteins	NN	O	O
,	NN	O	O
and	NN	O	O
total	NN	O	O
cholesterol	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
mice	NN	O	O
manifest	NN	O	O
several	NN	O	O
characteristics	NN	O	O
that	NN	O	O
are	NN	O	O
occasionally	NN	O	O
observed	NN	O	O
in	NN	O	O
human	NN	O	O
hypobetalipoproteinemia	NN	O	B-Disease
,	NN	O	O
including	NN	O	O
reduced	NN	O	O
plasma	NN	O	O
triglyceride	NN	O	O
concentrations	NN	O	O
,	NN	O	O
fasting	NN	O	O
chylomicronemia	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
reduced	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
cholesterol	NN	O	O
.	NN	O	O

An	NN	O	O
unexpected	NN	O	O
finding	NN	O	O
is	NN	O	O
that	NN	O	O
the	NN	O	O
modified	NN	O	O
Apob	NN	O	O
allele	NN	O	O
is	NN	O	O
strongly	NN	O	O
associated	NN	O	O
with	NN	O	O
exencephalus	NN	O	B-Disease
and	NN	O	O
hydrocephalus	NN	O	B-Disease
.	NN	O	O

These	NN	O	O
mice	NN	O	O
should	NN	O	O
help	NN	O	O
increase	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
hypobetalipoproteinemia	NN	O	B-Disease
,	NN	O	O
atherogenesis	NN	O	B-Disease
,	NN	O	O
and	NN	O	O
the	NN	O	O
etiology	NN	O	O
of	NN	O	O
exencephalus	NN	O	B-Disease
and	NN	O	O
hydrocephalus	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
disease	NN	O	O
with	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
mitochondrial	NN	O	I-Disease
very	NN	O	I-Disease
-	NN	O	I-Disease
long	NN	O	I-Disease
-	NN	O	I-Disease
chain	NN	O	I-Disease
acyl	NN	O	I-Disease
-	NN	O	I-Disease
CoA	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
.	NN	O	O

Palmitoyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
dehydrogenase	NN	O	O
activity	NN	O	O
in	NN	O	O
skin	NN	O	O
fibroblasts	NN	O	O
from	NN	O	O
seven	NN	O	O
patients	NN	O	O
with	NN	O	O
unidentified	NN	O	O
defects	NN	O	O
of	NN	O	O
fatty	NN	O	O
acid	NN	O	O
oxidation	NN	O	O
was	NN	O	O
measured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
antibodies	NN	O	O
against	NN	O	O
medium	NN	O	O
-	NN	O	O
chain	NN	O	O
,	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
,	NN	O	O
and	NN	O	O
very	NN	O	O
-	NN	O	O
long	NN	O	O
-	NN	O	O
chain	NN	O	O
acyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
dehydrogenases	NN	O	O
(	NN	O	O
VLCAD	NN	O	O
)	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
,	NN	O	O
4	NN	O	O
-	NN	O	O
5	NN	O	O
month	NN	O	O
old	NN	O	O
boys	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
novel	NN	O	O
disease	NN	O	O
,	NN	O	O
VLCAD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
as	NN	O	O
judged	NN	O	O
from	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
very	NN	O	O
low	NN	O	O
palmitoyl	NN	O	O
-	NN	O	O
CoA	NN	O	O
dehydrogenase	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
immunoreactivity	NN	O	O
toward	NN	O	O
antibody	NN	O	O
raised	NN	O	O
to	NN	O	O
purified	NN	O	O
VLCAD	NN	O	O
.	NN	O	O
.	NN	O	O

Molecular	NN	O	O
characterization	NN	O	O
of	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
patients	NN	O	O
of	NN	O	O
Chinese	NN	O	O
descent	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
new	NN	O	O
base	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
G6PD	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
underlying	NN	O	O
DNA	NN	O	O
changes	NN	O	O
associated	NN	O	O
with	NN	O	O
glucose	NN	O	B-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	I-Disease
G6PD	NN	O	I-Disease
)	NN	O	I-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
Asians	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
extensively	NN	O	O
investigated	NN	O	O
.	NN	O	O

To	NN	O	O
fill	NN	O	O
this	NN	O	O
gap	NN	O	O
,	NN	O	O
we	NN	O	O
sequenced	NN	O	O
the	NN	O	O
G6PD	NN	O	O
gene	NN	O	O
of	NN	O	O
43	NN	O	O
G6PD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
Chinese	NN	O	O
whose	NN	O	O
G6PD	NN	O	O
was	NN	O	O
well	NN	O	O
characterized	NN	O	O
biochemically	NN	O	O
.	NN	O	O

DNA	NN	O	O
samples	NN	O	O
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
these	NN	O	O
individuals	NN	O	O
for	NN	O	O
sequencing	NN	O	O
using	NN	O	O
a	NN	O	O
direct	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
sequencing	NN	O	O
procedure	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
43	NN	O	O
samples	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
five	NN	O	O
different	NN	O	O
types	NN	O	O
of	NN	O	O
nucleotide	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
G6PD	NN	O	O
gene	NN	O	O
at	NN	O	O
cDNA	NN	O	O
1388	NN	O	O
from	NN	O	O
G	NN	O	O
to	NN	O	O
A	NN	O	O
(	NN	O	O
Arg	NN	O	O
to	NN	O	O
His	NN	O	O
)	NN	O	O
;	NN	O	O
at	NN	O	O
cDNA	NN	O	O
1376	NN	O	O
from	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
(	NN	O	O
Arg	NN	O	O
to	NN	O	O
Leu	NN	O	O
)	NN	O	O
;	NN	O	O
at	NN	O	O
cDNA	NN	O	O
1024	NN	O	O
from	NN	O	O
C	NN	O	O
to	NN	O	O
T	NN	O	O
(	NN	O	O
Leu	NN	O	O
to	NN	O	O
Phe	NN	O	O
)	NN	O	O
;	NN	O	O
at	NN	O	O
cDNA	NN	O	O
392	NN	O	O
from	NN	O	O
G	NN	O	O
to	NN	O	O
T	NN	O	O
(	NN	O	O
Gly	NN	O	O
to	NN	O	O
Val	NN	O	O
)	NN	O	O
;	NN	O	O
at	NN	O	O
cDNA	NN	O	O
95	NN	O	O
from	NN	O	O
A	NN	O	O
to	NN	O	O
G	NN	O	O
(	NN	O	O
His	NN	O	O
to	NN	O	O
Arg	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
five	NN	O	O
nucleotide	NN	O	O
substitutions	NN	O	O
account	NN	O	O
for	NN	O	O
over	NN	O	O
83	NN	O	O
%	NN	O	O
of	NN	O	O
our	NN	O	O
43	NN	O	O
G6PD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
samples	NN	O	O
and	NN	O	O
these	NN	O	O
substitutions	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
non	NN	O	O
-	NN	O	O
Asians	NN	O	O
.	NN	O	O

The	NN	O	O
substitutions	NN	O	O
found	NN	O	O
at	NN	O	O
cDNA	NN	O	O
392	NN	O	O
and	NN	O	O
cDNA	NN	O	O
1024	NN	O	O
are	NN	O	O
new	NN	O	O
findings	NN	O	O
.	NN	O	O

The	NN	O	O
substitutions	NN	O	O
at	NN	O	O
cDNA	NN	O	O
1376	NN	O	O
and	NN	O	O
1388	NN	O	O
account	NN	O	O
for	NN	O	O
over	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
43	NN	O	O
samples	NN	O	O
examined	NN	O	O
indicating	NN	O	O
a	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
alleles	NN	O	O
among	NN	O	O
G6PD	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
Chinese	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
add	NN	O	O
support	NN	O	O
to	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
diverse	NN	O	O
point	NN	O	O
mutations	NN	O	O
may	NN	O	O
account	NN	O	O
largely	NN	O	O
for	NN	O	O
much	NN	O	O
of	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
Danish	NN	O	O
choroideremia	NN	O	B-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
searched	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
choroideremia	NN	O	B-Disease
gene	NN	O	O
(	NN	O	O
CHM	NN	O	B-Disease
)	NN	O	O
in	NN	O	O
patients	NN	O	O
from	NN	O	O
12	NN	O	O
Danish	NN	O	O
families	NN	O	O
in	NN	O	O
which	NN	O	O
CHM	NN	O	B-Disease
is	NN	O	O
segregating	NN	O	O
.	NN	O	O

Employing	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
,	NN	O	O
single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
direct	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
,	NN	O	O
different	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
6	NN	O	O
patients	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
mutations	NN	O	O
will	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
correct	NN	O	O
translation	NN	O	O
of	NN	O	O
the	NN	O	O
mRNA	NN	O	O
predicting	NN	O	O
a	NN	O	O
truncated	NN	O	O
protein	NN	O	O
or	NN	O	O
no	NN	O	O
gene	NN	O	O
product	NN	O	O
at	NN	O	O
all	NN	O	O
.	NN	O	O
.	NN	O	O

Structure	NN	O	O
and	NN	O	O
genomic	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
)	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
causing	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
DM	NN	O	B-Disease
)	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
unstable	NN	O	O
CTG	NN	O	O
trinucleotide	NN	O	O
repeat	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
encoding	NN	O	O
for	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
putative	NN	O	O
serine	NN	O	O
-	NN	O	O
threonine	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
present	NN	O	O
the	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
and	NN	O	O
murine	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
these	NN	O	O
sequences	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
and	NN	O	O
with	NN	O	O
known	NN	O	O
cDNA	NN	O	O
sequences	NN	O	O
from	NN	O	O
both	NN	O	O
species	NN	O	O
,	NN	O	O
led	NN	O	O
us	NN	O	O
to	NN	O	O
predict	NN	O	O
a	NN	O	O
translation	NN	O	O
initiation	NN	O	O
codon	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
determine	NN	O	O
the	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
.	NN	O	O

Several	NN	O	O
polymorphisms	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
and	NN	O	O
PCR	NN	O	O
assays	NN	O	O
to	NN	O	O
detect	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

The	NN	O	O
complete	NN	O	O
sequence	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
DM	NN	O	B-Disease
kinase	NN	O	O
gene	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
novel	NN	O	O
polymorphisms	NN	O	O
within	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
represent	NN	O	O
an	NN	O	O
important	NN	O	O
step	NN	O	O
in	NN	O	O
a	NN	O	O
further	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
genetics	NN	O	O
of	NN	O	O
myotonic	NN	O	B-Disease
dystrophy	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
biology	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O
.	NN	O	O

Autosomal	NN	O	O
recessive	NN	O	O
transmission	NN	O	O
of	NN	O	O
hemophilia	NN	O	B-Disease
A	NN	O	I-Disease
due	NN	O	O
to	NN	O	O
a	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
differential	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
bleeding	NN	O	I-Disease
disorders	NN	O	I-Disease
,	NN	O	O
hemophilia	NN	O	B-Disease
A	NN	O	I-Disease
and	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
disease	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
occasionally	NN	O	O
confounded	NN	O	O
by	NN	O	O
the	NN	O	O
close	NN	O	O
molecular	NN	O	O
relationship	NN	O	O
of	NN	O	O
coagulation	NN	O	O
factor	NN	O	O
VIII	NN	O	O
and	NN	O	O
von	NN	O	B-Disease
Willebrand	NN	O	I-Disease
factor	NN	O	O
(	NN	O	O
vWF	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
describes	NN	O	O
the	NN	O	O
autosomal	NN	O	O
inheritance	NN	O	O
of	NN	O	O
a	NN	O	O
hemophilia	NN	O	B-Disease
A	NN	O	I-Disease
phenotype	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
vWF	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
defective	NN	O	O
factor	NN	O	O
VIII	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
was	NN	O	O
a	NN	O	O
female	NN	O	O
patient	NN	O	O
with	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
factor	NN	O	O
VIII	NN	O	O
activity	NN	O	O
.	NN	O	O

Polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
amplification	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
were	NN	O	O
employed	NN	O	O
to	NN	O	O
examine	NN	O	O
exons	NN	O	O
encoding	NN	O	O
the	NN	O	O
putative	NN	O	O
factor	NN	O	O
VIII	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
vWF	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
homozygous	NN	O	O
for	NN	O	O
a	NN	O	O
single	NN	O	O
point	NN	O	O
mutation	NN	O	O
causing	NN	O	O
a	NN	O	O
Thr	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Met	NN	O	O
substitution	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
position	NN	O	O
28	NN	O	O
in	NN	O	O
the	NN	O	O
mature	NN	O	O
vWF	NN	O	O
subunit	NN	O	O
.	NN	O	O

The	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
was	NN	O	O
determined	NN	O	O
to	NN	O	O
be	NN	O	O
recessive	NN	O	O
because	NN	O	O
heterozygous	NN	O	O
family	NN	O	O
members	NN	O	O
were	NN	O	O
clinically	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Recombinant	NN	O	O
vWF	NN	O	O
containing	NN	O	O
the	NN	O	O
observed	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitution	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
COS	NN	O	O
-	NN	O	O
1	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
vWF	NN	O	O
was	NN	O	O
processed	NN	O	O
and	NN	O	O
secreted	NN	O	O
normally	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
functionally	NN	O	O
equivalent	NN	O	O
to	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
vWF	NN	O	O
in	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
platelets	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mutant	NN	O	O
failed	NN	O	O
to	NN	O	O
bind	NN	O	O
factor	NN	O	O
VIII	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
was	NN	O	O
functionally	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
observed	NN	O	O
hemophilia	NN	O	B-Disease
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
family	NN	O	O
we	NN	O	O
describe	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
recessive	NN	O	O
inheritance	NN	O	O
of	NN	O	O
a	NN	O	O
recently	NN	O	O
recognized	NN	O	O
class	NN	O	O
of	NN	O	O
genetic	NN	O	O
bleeding	NN	O	O
disorders	NN	O	O
,	NN	O	O
we	NN	O	O
call	NN	O	O
"	NN	O	O
autosomal	NN	O	O
hemophilia	NN	O	B-Disease
.	NN	O	O

"	NN	O	O
We	NN	O	O
conclude	NN	O	O
that	NN	O	O
vWF	NN	O	O
mutation	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
under	NN	O	O
recognized	NN	O	O
cause	NN	O	O
of	NN	O	O
hemophilia	NN	O	B-Disease
,	NN	O	O
especially	NN	O	O
in	NN	O	O
cases	NN	O	O
where	NN	O	O
the	NN	O	O
inheritance	NN	O	O
pattern	NN	O	O
is	NN	O	O
not	NN	O	O
consistent	NN	O	O
with	NN	O	O
X	NN	O	O
-	NN	O	O
linked	NN	O	O
transmission	NN	O	O
.	NN	O	O

Somatic	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
mutation	NN	O	O
in	NN	O	O
clear	NN	O	O
cell	NN	O	O
papillary	NN	O	B-Disease
cystadenoma	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
epididymis	NN	O	I-Disease
.	NN	O	O

Papillary	NN	O	B-Disease
cystadenoma	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
epididymis	NN	O	I-Disease
is	NN	O	O
an	NN	O	O
uncommon	NN	O	O
benign	NN	O	O
lesion	NN	O	O
that	NN	O	O
may	NN	O	O
occur	NN	O	O
sporadically	NN	O	O
or	NN	O	O
as	NN	O	O
a	NN	O	O
manifestation	NN	O	O
of	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
(	NN	O	I-Disease
VHL	NN	O	I-Disease
)	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

Neither	NN	O	O
immunohistochemical	NN	O	O
studies	NN	O	O
nor	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
for	NN	O	O
this	NN	O	O
lesion	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
describe	NN	O	O
two	NN	O	O
cases	NN	O	O
of	NN	O	O
clear	NN	O	O
cell	NN	O	O
papillary	NN	O	B-Disease
cystadenoma	NN	O	I-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
epididymis	NN	O	I-Disease
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
initially	NN	O	O
confused	NN	O	O
with	NN	O	O
metastatic	NN	O	B-Disease
renal	NN	O	I-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
.	NN	O	O

Both	NN	O	O
lesions	NN	O	O
showed	NN	O	O
positive	NN	O	O
immunohistochemical	NN	O	O
staining	NN	O	O
for	NN	O	O
low	NN	O	O
and	NN	O	O
intermediate	NN	O	O
molecular	NN	O	O
weight	NN	O	O
keratins	NN	O	O
(	NN	O	O
Cam	NN	O	O
5	NN	O	O
.	NN	O	O
2	NN	O	O
and	NN	O	O
AE1	NN	O	O
/	NN	O	O
AE3	NN	O	O
)	NN	O	O
,	NN	O	O
EMA	NN	O	O
,	NN	O	O
vimentin	NN	O	O
,	NN	O	O
alpha	NN	O	O
1	NN	O	O
-	NN	O	O
antitrypsin	NN	O	O
,	NN	O	O
and	NN	O	O
alpha	NN	O	O
1	NN	O	O
-	NN	O	O
antichymotrypsin	NN	O	O
.	NN	O	O

Each	NN	O	O
was	NN	O	O
negative	NN	O	O
for	NN	O	O
CEA	NN	O	O
.	NN	O	O

Because	NN	O	O
clear	NN	O	O
cell	NN	O	O
papillary	NN	O	B-Disease
cystadenoma	NN	O	I-Disease
is	NN	O	O
similar	NN	O	O
to	NN	O	O
renal	NN	O	B-Disease
cell	NN	O	I-Disease
carcinoma	NN	O	I-Disease
histologically	NN	O	O
,	NN	O	O
and	NN	O	O
because	NN	O	O
both	NN	O	O
occur	NN	O	O
as	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
von	NN	O	B-Disease
Hippel	NN	O	I-Disease
-	NN	O	I-Disease
Lindau	NN	O	I-Disease
disease	NN	O	I-Disease
complex	NN	O	O
,	NN	O	O
the	NN	O	O
authors	NN	O	O
analyzed	NN	O	O
both	NN	O	O
cases	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

A	NN	O	O
somatic	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
mutation	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
tumors	NN	O	B-Disease
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
followed	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
revealed	NN	O	O
a	NN	O	O
cytosine	NN	O	O
to	NN	O	O
thymine	NN	O	O
transition	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
694	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
replacement	NN	O	O
of	NN	O	O
an	NN	O	O
arginine	NN	O	O
with	NN	O	O
a	NN	O	O
stop	NN	O	O
codon	NN	O	O
after	NN	O	O
the	NN	O	O
sixth	NN	O	O
amino	NN	O	O
acid	NN	O	O
of	NN	O	O
exon	NN	O	O
3	NN	O	O
.	NN	O	O

As	NN	O	O
the	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
tumor	NN	O	B-Disease
suppressor	NN	O	O
gene	NN	O	O
,	NN	O	O
VHL	NN	O	B-Disease
gene	NN	O	O
mutations	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
tumorigenesis	NN	O	O
in	NN	O	O
sporadic	NN	O	O
cystadenomas	NN	O	B-Disease
of	NN	O	I-Disease
the	NN	O	I-Disease
epididymis	NN	O	I-Disease
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
WASP	NN	O	O
mutations	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
isolated	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
reveals	NN	O	O
allelic	NN	O	O
heterogeneity	NN	O	O
at	NN	O	O
the	NN	O	O
WAS	NN	O	B-Disease
locus	NN	O	O
.	NN	O	O

Mutation	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
recently	NN	O	O
isolated	NN	O	O
WASP	NN	O	O
protein	NN	O	O
has	NN	O	O
now	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
genetic	NN	O	B-Disease
defect	NN	O	I-Disease
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Wiskott	NN	O	I-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
a	NN	O	O
primary	NN	O	O
immunodeficiency	NN	O	B-Disease
disease	NN	O	I-Disease
associated	NN	O	O
with	NN	O	O
extensive	NN	O	O
phenotypic	NN	O	O
variability	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
WASP	NN	O	O
mutations	NN	O	O
responsible	NN	O	O
for	NN	O	O
WAS	NN	O	B-Disease
,	NN	O	O
we	NN	O	O
used	NN	O	O
PCR	NN	O	O
-	NN	O	O
SSCP	NN	O	O
analysis	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
WASP	NN	O	O
gene	NN	O	O
mutation	NN	O	O
in	NN	O	O
19	NN	O	O
unrelated	NN	O	O
boys	NN	O	O
with	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
classical	NN	O	O
or	NN	O	O
attenuated	NN	O	O
WAS	NN	O	B-Disease
or	NN	O	O
isolated	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
.	NN	O	O

All	NN	O	O
19	NN	O	O
patients	NN	O	O
had	NN	O	O
WASP	NN	O	O
mutations	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
initial	NN	O	O
three	NN	O	O
or	NN	O	O
terminal	NN	O	O
three	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
unique	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
missense	NN	O	O
mutation	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
substitution	NN	O	O
of	NN	O	O
the	NN	O	O
arginine	NN	O	O
at	NN	O	O
WAS	NN	O	B-Disease
codon	NN	O	O
86	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
three	NN	O	O
boys	NN	O	O
with	NN	O	O
severe	NN	O	O
WAS	NN	O	B-Disease
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
one	NN	O	O
boy	NN	O	O
presenting	NN	O	O
with	NN	O	O
thrombocytopenia	NN	O	B-Disease
alone	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
three	NN	O	O
mutations	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
isolated	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
patients	NN	O	O
leave	NN	O	O
the	NN	O	O
reading	NN	O	O
frame	NN	O	O
intact	NN	O	O
,	NN	O	O
about	NN	O	O
one	NN	O	O
-	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
alterations	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
severe	NN	O	O
and	NN	O	O
attenuated	NN	O	O
WAS	NN	O	B-Disease
patients	NN	O	O
result	NN	O	O
in	NN	O	O
frameshifted	NN	O	O
transcript	NN	O	O
and	NN	O	O
premature	NN	O	O
translation	NN	O	O
termination	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
therefore	NN	O	O
confirm	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
with	NN	O	O
WASP	NN	O	O
mutation	NN	O	O
and	NN	O	O
identify	NN	O	O
WASP	NN	O	O
mutation	NN	O	O
as	NN	O	O
a	NN	O	O
cause	NN	O	O
for	NN	O	O
isolated	NN	O	O
congenital	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
in	NN	O	O
males	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
WASP	NN	O	O
gene	NN	O	O
defects	NN	O	O
responsible	NN	O	O
for	NN	O	O
isolated	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
and	NN	O	O
other	NN	O	O
mild	NN	O	O
presentations	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
do	NN	O	O
not	NN	O	O
appear	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
resulting	NN	O	O
in	NN	O	O
severe	NN	O	O
WAS	NN	O	B-Disease
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
analysis	NN	O	O
of	NN	O	O
WASP	NN	O	O
gene	NN	O	O
mutation	NN	O	O
provides	NN	O	O
a	NN	O	O
valuable	NN	O	O
tool	NN	O	O
for	NN	O	O
distinguishing	NN	O	O
the	NN	O	O
spectrum	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
patients	NN	O	O
and	NN	O	O
the	NN	O	O
subset	NN	O	O
of	NN	O	O
males	NN	O	O
with	NN	O	O
isolated	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
who	NN	O	O
represent	NN	O	O
mild	NN	O	O
cases	NN	O	O
of	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

WASP	NN	O	O
gene	NN	O	O
mutations	NN	O	O
in	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
WASP	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
cloned	NN	O	O
from	NN	O	O
Xp11	NN	O	O
.	NN	O	O

23	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
mutated	NN	O	O
in	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
Wiskott	NN	O	B-Disease
-	NN	O	I-Disease
Aldrich	NN	O	I-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
WAS	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
screening	NN	O	O
protocol	NN	O	O
for	NN	O	O
identifying	NN	O	O
WASP	NN	O	O
gene	NN	O	O
alterations	NN	O	O
in	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
and	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
spectrum	NN	O	O
of	NN	O	O
novel	NN	O	O
mutations	NN	O	O
in	NN	O	O
12	NN	O	O
additional	NN	O	O
unrelated	NN	O	O
families	NN	O	O
.	NN	O	O

These	NN	O	O
missense	NN	O	O
,	NN	O	O
nonsense	NN	O	O
and	NN	O	O
frameshift	NN	O	O
mutations	NN	O	O
involve	NN	O	O
eight	NN	O	O
of	NN	O	O
the	NN	O	O
12	NN	O	O
exons	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Two	NN	O	O
mutations	NN	O	O
creating	NN	O	O
premature	NN	O	O
termination	NN	O	O
codons	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
lack	NN	O	O
of	NN	O	O
detectable	NN	O	O
mRNA	NN	O	O
on	NN	O	O
Northern	NN	O	O
blots	NN	O	O
.	NN	O	O

Four	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
,	NN	O	O
Leu27Phe	NN	O	O
,	NN	O	O
Thr48Ile	NN	O	O
,	NN	O	O
Val75Met	NN	O	O
and	NN	O	O
Arg477Lys	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
congenital	NN	O	B-Disease
thrombocytopenia	NN	O	I-Disease
and	NN	O	O
no	NN	O	O
clinically	NN	O	O
evident	NN	O	O
immune	NN	O	O
defect	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
WASP	NN	O	O
gene	NN	O	O
is	NN	O	O
the	NN	O	O
site	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
thrombocytopenia	NN	O	I-Disease
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
WAS	NN	O	B-Disease
.	NN	O	O

A	NN	O	O
T	NN	O	O
-	NN	O	O
cell	NN	O	O
line	NN	O	O
from	NN	O	O
a	NN	O	O
WAS	NN	O	B-Disease
patient	NN	O	O
contained	NN	O	O
two	NN	O	O
independent	NN	O	O
DNA	NN	O	O
alterations	NN	O	O
,	NN	O	O
a	NN	O	O
constitutional	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
,	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
and	NN	O	O
compensatory	NN	O	O
splice	NN	O	O
site	NN	O	O
mutation	NN	O	O
unique	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

The	NN	O	O
distribution	NN	O	O
of	NN	O	O
eight	NN	O	O
missense	NN	O	O
mutations	NN	O	O
provides	NN	O	O
valuable	NN	O	O
information	NN	O	O
on	NN	O	O
amino	NN	O	O
acids	NN	O	O
which	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
protein	NN	O	O
function	NN	O	O
,	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
two	NN	O	O
exons	NN	O	O
are	NN	O	O
hot	NN	O	O
-	NN	O	O
spots	NN	O	O
for	NN	O	O
mutation	NN	O	O
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
inter	NN	O	O
-	NN	O	O
generational	NN	O	O
instability	NN	O	O
in	NN	O	O
the	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
and	NN	O	O
for	NN	O	O
conserved	NN	O	O
haplotypes	NN	O	O
at	NN	O	O
flanking	NN	O	O
markers	NN	O	O
amongst	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
subjects	NN	O	O
with	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
(	NN	O	O
CAG	NN	O	O
)	NN	O	O
n	NN	O	O
repeat	NN	O	O
array	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
'	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
and	NN	O	O
the	NN	O	O
haplotype	NN	O	O
at	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
microsatellite	NN	O	O
markers	NN	O	O
surrounding	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
cohort	NN	O	O
of	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
subjects	NN	O	O
affected	NN	O	O
with	NN	O	O
Machado	NN	O	B-Disease
-	NN	O	I-Disease
Joseph	NN	O	I-Disease
disease	NN	O	I-Disease
(	NN	O	O
MJD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
five	NN	O	O
novel	NN	O	O
observations	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
MJD	NN	O	B-Disease
is	NN	O	O
associated	NN	O	O
with	NN	O	O
expansion	NN	O	O
fo	NN	O	O
the	NN	O	O
array	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
of	NN	O	O
14	NN	O	O
-	NN	O	O
37	NN	O	O
repeats	NN	O	O
to	NN	O	O
68	NN	O	O
-	NN	O	O
84	NN	O	O
repeats	NN	O	O
in	NN	O	O
most	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
subjects	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
subjects	NN	O	O
were	NN	O	O
observed	NN	O	O
with	NN	O	O
expansions	NN	O	O
intermediate	NN	O	O
in	NN	O	O
size	NN	O	O
between	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
MJD	NN	O	B-Disease
affected	NN	O	O
groups	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
the	NN	O	O
expanded	NN	O	O
allele	NN	O	O
associated	NN	O	O
with	NN	O	O
MJD	NN	O	B-Disease
displays	NN	O	O
inter	NN	O	O
-	NN	O	O
generational	NN	O	O
instability	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
male	NN	O	O
meioses	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
instability	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
clinical	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
anticipation	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
expanded	NN	O	O
allele	NN	O	O
is	NN	O	O
not	NN	O	O
only	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
age	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
onset	NN	O	O
of	NN	O	O
MJD	NN	O	B-Disease
(	NN	O	O
r	NN	O	O
=	NN	O	O
-	NN	O	O
0	NN	O	O
.	NN	O	O
738	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
001	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
also	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
other	NN	O	O
clinical	NN	O	O
features	NN	O	O
[	NN	O	O
e	NN	O	O
.	NN	O	O
g	NN	O	O
.	NN	O	O
pseudoexophthalmos	NN	O	O
and	NN	O	O
pyramidal	NN	O	O
signs	NN	O	O
were	NN	O	O
more	NN	O	O
frequent	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
large	NN	O	O
repeats	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
001	NN	O	O
and	NN	O	O
p	NN	O	O
<	NN	O	O
0	NN	O	O
.	NN	O	O
05	NN	O	O
respectively	NN	O	O
)	NN	O	O
]	NN	O	O
.	NN	O	O

Fourth	NN	O	O
,	NN	O	O
the	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
is	NN	O	O
significantly	NN	O	O
more	NN	O	O
severe	NN	O	O
and	NN	O	O
had	NN	O	O
an	NN	O	O
early	NN	O	O
age	NN	O	O
of	NN	O	O
onset	NN	O	O
(	NN	O	O
16	NN	O	O
years	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
subject	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
expanded	NN	O	O
allele	NN	O	O
,	NN	O	O
which	NN	O	O
contrasts	NN	O	O
with	NN	O	O
Huntington	NN	O	B-Disease
disease	NN	O	I-Disease
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
expanded	NN	O	O
allele	NN	O	O
in	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
could	NN	O	O
exert	NN	O	O
its	NN	O	O
effect	NN	O	O
either	NN	O	O
by	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
(	NN	O	O
putatively	NN	O	O
excluded	NN	O	O
in	NN	O	O
HD	NN	O	B-Disease
)	NN	O	O
or	NN	O	O
by	NN	O	O
a	NN	O	O
gain	NN	O	O
of	NN	O	O
function	NN	O	O
effect	NN	O	O
as	NN	O	O
proposed	NN	O	O
for	NN	O	O
HD	NN	O	B-Disease
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
subjects	NN	O	O
affected	NN	O	O
with	NN	O	O
MJD	NN	O	B-Disease
share	NN	O	O
haplotypes	NN	O	O
at	NN	O	O
several	NN	O	O
markers	NN	O	O
surrounding	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
uncommon	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
Japanese	NN	O	O
and	NN	O	O
Caucasian	NN	O	O
population	NN	O	O
,	NN	O	O
and	NN	O	O
which	NN	O	O
suggests	NN	O	O
the	NN	O	O
existence	NN	O	O
either	NN	O	O
of	NN	O	O
common	NN	O	O
founders	NN	O	O
in	NN	O	O
these	NN	O	O
populations	NN	O	O
or	NN	O	O
of	NN	O	O
chromosomes	NN	O	O
susceptible	NN	O	O
to	NN	O	O
pathologic	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
CAG	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
MJD1	NN	O	O
gene	NN	O	O
.	NN	O	O

A	NN	O	O
4	NN	O	O
-	NN	O	O
megabase	NN	O	O
YAC	NN	O	O
contig	NN	O	O
that	NN	O	O
spans	NN	O	O
the	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
region	NN	O	O
on	NN	O	O
human	NN	O	O
chromosome	NN	O	O
8q24	NN	O	O
.	NN	O	O
1	NN	O	O
:	NN	O	O
use	NN	O	O
in	NN	O	O
refining	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
trichorhinophalangeal	NN	O	B-Disease
syndrome	NN	O	I-Disease
and	NN	O	O
multiple	NN	O	O
exostoses	NN	O	O
genes	NN	O	O
(	NN	O	O
TRPS1	NN	O	O
and	NN	O	O
EXT1	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
constructed	NN	O	O
a	NN	O	O
physical	NN	O	O
map	NN	O	O
covering	NN	O	O
over	NN	O	O
4	NN	O	O
Mb	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
8q24	NN	O	O
.	NN	O	O

1	NN	O	O
and	NN	O	O
used	NN	O	O
this	NN	O	O
map	NN	O	O
to	NN	O	O
refine	NN	O	O
the	NN	O	O
locations	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
responsible	NN	O	O
for	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
map	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
overlapping	NN	O	O
YAC	NN	O	O
clones	NN	O	O
that	NN	O	O
were	NN	O	O
identified	NN	O	O
and	NN	O	O
ordered	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
sequence	NN	O	O
tagged	NN	O	O
sites	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
Langer	NN	O	O
-	NN	O	O
Giedion	NN	O	O
chromosomal	NN	O	O
region	NN	O	O
on	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
.	NN	O	O

The	NN	O	O
minimal	NN	O	O
region	NN	O	O
of	NN	O	O
overlap	NN	O	O
of	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
deletions	NN	O	O
,	NN	O	O
previously	NN	O	O
identified	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
15	NN	O	O
patients	NN	O	O
,	NN	O	O
was	NN	O	O
placed	NN	O	O
on	NN	O	O
the	NN	O	O
map	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
2	NN	O	O
patients	NN	O	O
whose	NN	O	O
deletions	NN	O	O
define	NN	O	O
the	NN	O	O
endpoints	NN	O	O
.	NN	O	O

The	NN	O	O
chromosome	NN	O	O
8	NN	O	O
breakpoint	NN	O	O
of	NN	O	O
a	NN	O	O
balanced	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
9	NN	O	O
)	NN	O	O
(	NN	O	O
q24	NN	O	O
.	NN	O	O
11	NN	O	O
;	NN	O	O
q33	NN	O	O
.	NN	O	O
3	NN	O	O
)	NN	O	O
translocation	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
trichorhinophalangeal	NN	O	B-Disease
syndrome	NN	O	I-Disease
(	NN	O	O
TRPS	NN	O	B-Disease
I	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
located	NN	O	O
just	NN	O	O
within	NN	O	O
the	NN	O	O
proximal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
minimal	NN	O	O
deletion	NN	O	O
region	NN	O	O
.	NN	O	O

A	NN	O	O
deletion	NN	O	O
of	NN	O	O
8q24	NN	O	O
.	NN	O	O

11	NN	O	O
-	NN	O	O
q24	NN	O	O
.	NN	O	O

3	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
multiple	NN	O	O
exostoses	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
overlap	NN	O	O
the	NN	O	O
distal	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
LGS	NN	O	B-Disease
deletion	NN	O	O
region	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
EXT1	NN	O	O
gene	NN	O	O
is	NN	O	O
distal	NN	O	O
to	NN	O	O
the	NN	O	O
TRPS1	NN	O	O
gene	NN	O	O
and	NN	O	O
supporting	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
Langer	NN	O	B-Disease
-	NN	O	I-Disease
Giedion	NN	O	I-Disease
syndrome	NN	O	I-Disease
is	NN	O	O
due	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
functional	NN	O	O
copies	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
TRPS1	NN	O	O
and	NN	O	O
the	NN	O	O
EXT1	NN	O	O
genes	NN	O	O

BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
sample	NN	O	O
of	NN	O	O
young	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

BACKGROUND	NN	O	O
.	NN	O	O

Inherited	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
risk	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
in	NN	O	O
some	NN	O	O
families	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
DNA	NN	O	O
samples	NN	O	O
from	NN	O	O
women	NN	O	O
enrolled	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
-	NN	O	O
based	NN	O	O
study	NN	O	O
of	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
to	NN	O	O
assess	NN	O	O
the	NN	O	O
spectrum	NN	O	O
and	NN	O	O
frequency	NN	O	O
of	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
young	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
80	NN	O	O
women	NN	O	O
in	NN	O	O
whom	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
was	NN	O	O
diagnosed	NN	O	O
before	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
35	NN	O	O
,	NN	O	O
and	NN	O	O
who	NN	O	O
were	NN	O	O
not	NN	O	O
selected	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
family	NN	O	O
history	NN	O	O
.	NN	O	O

Genomic	NN	O	O
DNA	NN	O	O
was	NN	O	O
studied	NN	O	O
for	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
by	NN	O	O
analysis	NN	O	O
involving	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphisms	NN	O	O
and	NN	O	O
with	NN	O	O
allele	NN	O	O
-	NN	O	O
specific	NN	O	O
assays	NN	O	O
.	NN	O	O

Alterations	NN	O	O
were	NN	O	O
defined	NN	O	O
by	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Germ	NN	O	O
-	NN	O	O
line	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
6	NN	O	O
of	NN	O	O
the	NN	O	O
80	NN	O	O
women	NN	O	O
.	NN	O	O

Four	NN	O	O
additional	NN	O	O
rare	NN	O	O
sequence	NN	O	O
variants	NN	O	O
of	NN	O	O
unknown	NN	O	O
functional	NN	O	O
importance	NN	O	O
were	NN	O	O
also	NN	O	O
identified	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
and	NN	O	O
three	NN	O	O
of	NN	O	O
the	NN	O	O
rare	NN	O	O
sequence	NN	O	O
variants	NN	O	O
were	NN	O	O
found	NN	O	O
among	NN	O	O
the	NN	O	O
39	NN	O	O
women	NN	O	O
who	NN	O	O
reported	NN	O	O
no	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
and	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
rare	NN	O	O
variants	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
reference	NN	O	O
population	NN	O	O
of	NN	O	O
73	NN	O	O
unrelated	NN	O	O
subjects	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
.	NN	O	O

Alterations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
approximately	NN	O	O
10	NN	O	O
percent	NN	O	O
of	NN	O	O
this	NN	O	O
cohort	NN	O	O
of	NN	O	O
young	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
risk	NN	O	O
of	NN	O	O
harboring	NN	O	O
a	NN	O	O
mutation	NN	O	O
was	NN	O	O
not	NN	O	O
limited	NN	O	O
to	NN	O	O
women	NN	O	O
with	NN	O	O
family	NN	O	O
histories	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
or	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

These	NN	O	O
results	NN	O	O
represent	NN	O	O
a	NN	O	O
minimal	NN	O	O
estimate	NN	O	O
of	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

Comprehensive	NN	O	O
methods	NN	O	O
of	NN	O	O
identifying	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
and	NN	O	O
understanding	NN	O	O
their	NN	O	O
importance	NN	O	O
will	NN	O	O
be	NN	O	O
needed	NN	O	O
before	NN	O	O
testing	NN	O	O
of	NN	O	O
women	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
can	NN	O	O
be	NN	O	O
undertaken	NN	O	O
.	NN	O	O
.	NN	O	O

Novel	NN	O	O
inherited	NN	O	O
mutations	NN	O	O
and	NN	O	O
variable	NN	O	O
expressivity	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
alleles	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
founder	NN	O	O
mutation	NN	O	O
185delAG	NN	O	O
in	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
families	NN	O	O
.	NN	O	O

Thirty	NN	O	O
-	NN	O	O
seven	NN	O	O
families	NN	O	O
with	NN	O	O
four	NN	O	O
or	NN	O	O
more	NN	O	O
cases	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
or	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Twelve	NN	O	O
different	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
,	NN	O	O
four	NN	O	O
novel	NN	O	O
and	NN	O	O
eight	NN	O	O
previously	NN	O	O
observed	NN	O	O
,	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
16	NN	O	O
families	NN	O	O
.	NN	O	O

Five	NN	O	O
families	NN	O	O
of	NN	O	O
Ashkenazi	NN	O	O
Jewish	NN	O	O
descent	NN	O	O
carried	NN	O	O
the	NN	O	O
185delAG	NN	O	O
mutation	NN	O	O
and	NN	O	O
shared	NN	O	O
the	NN	O	O
same	NN	O	O
haplotype	NN	O	O
at	NN	O	O
eight	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
spanning	NN	O	O
approximately	NN	O	O
850	NN	O	O
kb	NN	O	O
at	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Expressivity	NN	O	O
of	NN	O	O
185delAG	NN	O	O
in	NN	O	O
these	NN	O	O
families	NN	O	O
varied	NN	O	O
,	NN	O	O
from	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
without	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Mutation	NN	O	O
4184delTCAA	NN	O	O
occurred	NN	O	O
independently	NN	O	O
in	NN	O	O
two	NN	O	O
families	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
family	NN	O	O
,	NN	O	O
penetrance	NN	O	O
was	NN	O	O
complete	NN	O	O
,	NN	O	O
with	NN	O	O
females	NN	O	O
developing	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
or	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
and	NN	O	O
the	NN	O	O
male	NN	O	O
carrier	NN	O	O
developing	NN	O	O
prostatic	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
whereas	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
family	NN	O	O
,	NN	O	O
penetrance	NN	O	O
was	NN	O	O
incomplete	NN	O	O
and	NN	O	O
only	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
occurred	NN	O	O
,	NN	O	O
diagnosed	NN	O	O
at	NN	O	O
ages	NN	O	O
38	NN	O	O
-	NN	O	O
81	NN	O	O
years	NN	O	O
.	NN	O	O

Two	NN	O	O
novel	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
mutant	NN	O	O
BRCA1	NN	O	O
transcript	NN	O	O
in	NN	O	O
families	NN	O	O
with	NN	O	O
10	NN	O	O
and	NN	O	O
6	NN	O	O
cases	NN	O	O
of	NN	O	O
early	NN	O	O
-	NN	O	O
onset	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
and	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
.	NN	O	O

A	NN	O	O
665	NN	O	O
-	NN	O	O
nt	NN	O	O
segment	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
3	NN	O	O
-	NN	O	O
UTR	NN	O	O
and	NN	O	O
1	NN	O	O
.	NN	O	O

3	NN	O	O
kb	NN	O	O
of	NN	O	O
genomic	NN	O	O
sequence	NN	O	O
including	NN	O	O
the	NN	O	O
putative	NN	O	O
promoter	NN	O	O
region	NN	O	O
were	NN	O	O
invariant	NN	O	O
by	NN	O	O
single	NN	O	O
-	NN	O	O
strand	NN	O	O
conformation	NN	O	O
analysis	NN	O	O
in	NN	O	O
13	NN	O	O
families	NN	O	O
without	NN	O	O
coding	NN	O	O
-	NN	O	O
sequence	NN	O	O
mutations	NN	O	O
.	NN	O	O

Overall	NN	O	O
in	NN	O	O
our	NN	O	O
series	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
26	NN	O	O
families	NN	O	O
16	NN	O	O
with	NN	O	O
positive	NN	O	O
BRCA1	NN	O	O
lod	NN	O	O
scores	NN	O	O
,	NN	O	O
7	NN	O	O
with	NN	O	O
negative	NN	O	O
lod	NN	O	O
scores	NN	O	O
(	NN	O	O
reflecting	NN	O	O
multiple	NN	O	O
sporadic	NN	O	O
breast	NN	O	B-Disease
cancers	NN	O	I-Disease
)	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
not	NN	O	O
tested	NN	O	O
for	NN	O	O
linkage	NN	O	O
.	NN	O	O

Three	NN	O	O
other	NN	O	O
families	NN	O	O
have	NN	O	O
positive	NN	O	O
lod	NN	O	O
scores	NN	O	O
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
BRCA2	NN	O	O
,	NN	O	O
but	NN	O	O
13	NN	O	O
families	NN	O	O
without	NN	O	O
detected	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
have	NN	O	O
negative	NN	O	O
lod	NN	O	O
scores	NN	O	O
for	NN	O	O
both	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
BRCA2	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
glucose	NN	O	O
-	NN	O	O
6	NN	O	O
-	NN	O	O
phosphate	NN	O	O
dehydrogenase	NN	O	O
variant	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Orissa	NN	O	O
(	NN	O	O
44	NN	O	O
Ala	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Gly	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
polymorphic	NN	O	O
variant	NN	O	O
in	NN	O	O
tribal	NN	O	O
populations	NN	O	O
in	NN	O	O
India	NN	O	O
.	NN	O	O

Deficiency	NN	O	B-Disease
of	NN	O	I-Disease
glucose	NN	O	I-Disease
-	NN	O	I-Disease
6	NN	O	I-Disease
-	NN	O	I-Disease
phosphate	NN	O	I-Disease
dehydrogenase	NN	O	I-Disease
(	NN	O	O
G6PD	NN	O	O
)	NN	O	O
is	NN	O	O
usually	NN	O	O
found	NN	O	O
at	NN	O	O
high	NN	O	O
frequencies	NN	O	O
in	NN	O	O
areas	NN	O	O
of	NN	O	O
the	NN	O	O
world	NN	O	O
where	NN	O	O
malaria	NN	O	B-Disease
has	NN	O	O
been	NN	O	O
endemic	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
and	NN	O	O
genetic	NN	O	O
basis	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
have	NN	O	O
been	NN	O	O
studied	NN	O	O
in	NN	O	O
Africa	NN	O	O
,	NN	O	O
around	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
Far	NN	O	O
East	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
such	NN	O	O
information	NN	O	O
is	NN	O	O
available	NN	O	O
about	NN	O	O
the	NN	O	O
situation	NN	O	O
in	NN	O	O
India	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
G6PD	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
several	NN	O	O
different	NN	O	O
Indian	NN	O	O
populations	NN	O	O
by	NN	O	O
screening	NN	O	O
for	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
,	NN	O	O
followed	NN	O	O
by	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
deficient	NN	O	O
alleles	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
varies	NN	O	O
between	NN	O	O
3	NN	O	O
%	NN	O	O
and	NN	O	O
15	NN	O	O
%	NN	O	O
in	NN	O	O
different	NN	O	O
tribal	NN	O	O
and	NN	O	O
urban	NN	O	O
groups	NN	O	O
.	NN	O	O

Remarkably	NN	O	O
,	NN	O	O
a	NN	O	O
previously	NN	O	O
unreported	NN	O	O
deficient	NN	O	O
variant	NN	O	O
,	NN	O	O
G6PD	NN	O	O
Orissa	NN	O	O
(	NN	O	O
44	NN	O	O
Ala	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Gly	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
in	NN	O	O
tribal	NN	O	O
Indian	NN	O	O
populations	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
urban	NN	O	O
populations	NN	O	O
,	NN	O	O
where	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
G6PD	NN	O	B-Disease
deficiency	NN	O	I-Disease
is	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
G6PD	NN	O	O
Mediterranean	NN	O	O
(	NN	O	O
188	NN	O	O
Ser	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
Phe	NN	O	O
)	NN	O	O
variant	NN	O	O
.	NN	O	O

The	NN	O	O
KmNADP	NN	O	O
of	NN	O	O
G6PD	NN	O	O
Orissa	NN	O	O
is	NN	O	O
fivefold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
enzyme	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
the	NN	O	O
alanine	NN	O	O
residue	NN	O	O
that	NN	O	O
is	NN	O	O
replaced	NN	O	O
by	NN	O	O
glycine	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	O
coenzyme	NN	O	O
-	NN	O	O
binding	NN	O	O
site	NN	O	O
.	NN	O	O
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
of	NN	O	O
bipolar	NN	O	B-Disease
disorder	NN	O	I-Disease
to	NN	O	O
chromosome	NN	O	O
18	NN	O	O
with	NN	O	O
a	NN	O	O
parent	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
origin	NN	O	O
effect	NN	O	O
.	NN	O	O

A	NN	O	O
susceptibility	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	O
18	NN	O	O
and	NN	O	O
a	NN	O	O
parent	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
origin	NN	O	O
effect	NN	O	O
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
for	NN	O	O
bipolar	NN	O	B-Disease
affective	NN	O	I-Disease
disorder	NN	O	I-Disease
(	NN	O	O
BPAD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
28	NN	O	O
nuclear	NN	O	O
families	NN	O	O
selected	NN	O	O
for	NN	O	O
apparent	NN	O	O
unilineal	NN	O	O
transmission	NN	O	O
of	NN	O	O
the	NN	O	O
BPAD	NN	O	B-Disease
phenotype	NN	O	O
,	NN	O	O
by	NN	O	O
using	NN	O	O
31	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
spanning	NN	O	O
chromosome	NN	O	O
18	NN	O	O
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
was	NN	O	O
tested	NN	O	O
with	NN	O	O
affected	NN	O	O
-	NN	O	O
sib	NN	O	O
-	NN	O	O
pair	NN	O	O
and	NN	O	O
LOD	NN	O	O
score	NN	O	O
methods	NN	O	O
under	NN	O	O
two	NN	O	O
definitions	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
affected	NN	O	O
-	NN	O	O
sibpair	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
excess	NN	O	O
allele	NN	O	O
sharing	NN	O	O
for	NN	O	O
markers	NN	O	O
on	NN	O	O
18p	NN	O	O
within	NN	O	O
the	NN	O	O
region	NN	O	O
reported	NN	O	O
previously	NN	O	O
.	NN	O	O

The	NN	O	O
greatest	NN	O	O
sharing	NN	O	O
was	NN	O	O
at	NN	O	O
D18S37	NN	O	O
64	NN	O	O
%	NN	O	O
in	NN	O	O
bipolar	NN	O	O
and	NN	O	O
recurrent	NN	O	O
unipolar	NN	O	O
(	NN	O	O
RUP	NN	O	O
)	NN	O	O
sib	NN	O	O
pairs	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
0006	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
excess	NN	O	O
sharing	NN	O	O
of	NN	O	O
the	NN	O	O
paternally	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
maternally	NN	O	O
,	NN	O	O
transmitted	NN	O	O
alleles	NN	O	O
was	NN	O	O
observed	NN	O	O
at	NN	O	O
three	NN	O	O
markers	NN	O	O
on	NN	O	O
18q	NN	O	O
at	NN	O	O
D18S41	NN	O	O
,	NN	O	O
51	NN	O	O
bipolar	NN	O	O
and	NN	O	O
RUP	NN	O	O
sib	NN	O	O
pairs	NN	O	O
were	NN	O	O
concordant	NN	O	O
for	NN	O	O
paternally	NN	O	O
transmitted	NN	O	O
alleles	NN	O	O
,	NN	O	O
and	NN	O	O
21	NN	O	O
pairs	NN	O	O
were	NN	O	O
discordant	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0004	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
evidence	NN	O	O
for	NN	O	O
linkage	NN	O	O
to	NN	O	O
loci	NN	O	O
on	NN	O	O
both	NN	O	O
18p	NN	O	O
and	NN	O	O
18q	NN	O	O
was	NN	O	O
strongest	NN	O	O
in	NN	O	O
the	NN	O	O
11	NN	O	O
paternal	NN	O	O
pedigrees	NN	O	O
,	NN	O	O
i	NN	O	O
.	NN	O	O

e	NN	O	O
e	NN	O	O
.	NN	O	O
,	NN	O	O
those	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
father	NN	O	O
or	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
fathers	NN	O	O
sibs	NN	O	O
is	NN	O	O
affected	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
pedigrees	NN	O	O
,	NN	O	O
the	NN	O	O
greatest	NN	O	O
allele	NN	O	O
sharing	NN	O	O
(	NN	O	O
81	NN	O	O
%	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
00002	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
highest	NN	O	O
LOD	NN	O	O
score	NN	O	O
(	NN	O	O
3	NN	O	O
.	NN	O	O
51	NN	O	O
;	NN	O	O
phi	NN	O	O
=	NN	O	O
0	NN	O	O
.	NN	O	O
0	NN	O	O
)	NN	O	O
were	NN	O	O
observed	NN	O	O
at	NN	O	O
D18S41	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
further	NN	O	O
support	NN	O	O
for	NN	O	O
linkage	NN	O	O
of	NN	O	O
BPAD	NN	O	B-Disease
to	NN	O	O
chromosome	NN	O	O
18	NN	O	O
and	NN	O	O
the	NN	O	O
first	NN	O	O
molecular	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
parent	NN	O	O
-	NN	O	O
of	NN	O	O
-	NN	O	O
origin	NN	O	O
effect	NN	O	O
operating	NN	O	O
in	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
loci	NN	O	O
involved	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
precise	NN	O	O
location	NN	O	O
,	NN	O	O
require	NN	O	O
further	NN	O	O
study	NN	O	O

A	NN	O	O
prevalent	NN	O	O
mutation	NN	O	O
for	NN	O	O
galactosemia	NN	O	B-Disease
among	NN	O	O
black	NN	O	O
Americans	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
To	NN	O	O
define	NN	O	O
the	NN	O	O
mutation	NN	O	O
causing	NN	O	O
galactosemia	NN	O	B-Disease
in	NN	O	O
patients	NN	O	O
of	NN	O	O
black	NN	O	O
American	NN	O	O
origin	NN	O	O
who	NN	O	O
have	NN	O	O
no	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyltransferase	NN	O	O
(	NN	O	O
GALT	NN	O	O
)	NN	O	O
activity	NN	O	O
in	NN	O	O
erythrocytes	NN	O	O
but	NN	O	O
good	NN	O	O
clinical	NN	O	O
outcome	NN	O	O
.	NN	O	O

METHODS	NN	O	O
We	NN	O	O
discovered	NN	O	O
a	NN	O	O
mutation	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
C	NN	O	O
-	NN	O	O
-	NN	O	O
>	NN	O	O
T	NN	O	O
transition	NN	O	O
at	NN	O	O
base	NN	O	O
-	NN	O	O
pair	NN	O	O
1158	NN	O	O
of	NN	O	O
the	NN	O	O
GALT	NN	O	O
gene	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
serine	NN	O	O
-	NN	O	O
to	NN	O	O
-	NN	O	O
leucine	NN	O	O
substitution	NN	O	O
at	NN	O	O
codon	NN	O	O
135	NN	O	O
(	NN	O	O
S135L	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
developed	NN	O	O
a	NN	O	O
method	NN	O	O
with	NN	O	O
which	NN	O	O
to	NN	O	O
screen	NN	O	O
populations	NN	O	O
for	NN	O	O
its	NN	O	O
prevalence	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
galactose	NN	O	O
-	NN	O	O
1	NN	O	O
-	NN	O	O
phosphate	NN	O	O
uridyltransferase	NN	O	O
among	NN	O	O
erythrocytes	NN	O	O
,	NN	O	O
leukocytes	NN	O	O
,	NN	O	O
and	NN	O	O
transformed	NN	O	O
lymphoblasts	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
total	NN	O	O
body	NN	O	O
oxidation	NN	O	O
of	NN	O	O
D	NN	O	O
-	NN	O	O
(	NN	O	O
13C	NN	O	O
)	NN	O	O
-	NN	O	O
galactose	NN	O	O
to	NN	O	O
13CO2	NN	O	O
among	NN	O	O
three	NN	O	O
genotypes	NN	O	O
for	NN	O	O
GALT	NN	O	O
(	NN	O	O
S135L	NN	O	O
/	NN	O	O
S135L	NN	O	O
,	NN	O	O
Q188R	NN	O	O
/	NN	O	O
Q188R	NN	O	O
,	NN	O	O
and	NN	O	O
Normal	NN	O	O
/	NN	O	O
Normal	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
We	NN	O	O
found	NN	O	O
a	NN	O	O
48	NN	O	O
%	NN	O	O
prevalence	NN	O	O
of	NN	O	O
the	NN	O	O
S135L	NN	O	O
mutation	NN	O	O
among	NN	O	O
17	NN	O	O
black	NN	O	O
American	NN	O	O
patients	NN	O	O
with	NN	O	O
classic	NN	O	B-Disease
galactosemia	NN	O	I-Disease
and	NN	O	O
a	NN	O	O
1	NN	O	O
%	NN	O	O
prevalence	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
50	NN	O	O
black	NN	O	O
Americans	NN	O	O
without	NN	O	O
galactosemia	NN	O	B-Disease
.	NN	O	O

The	NN	O	O
S135L	NN	O	O
mutation	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
84	NN	O	O
white	NN	O	O
patients	NN	O	O
with	NN	O	O
G	NN	O	O
/	NN	O	O
G	NN	O	O
galactosemia	NN	O	B-Disease
nor	NN	O	O
in	NN	O	O
87	NN	O	O
white	NN	O	O
control	NN	O	O
subjects	NN	O	O
without	NN	O	O
galactosemia	NN	O	B-Disease
.	NN	O	O

We	NN	O	O
found	NN	O	O
normal	NN	O	O
whole	NN	O	O
body	NN	O	O
oxidation	NN	O	O
of	NN	O	O
D	NN	O	O
-	NN	O	O
(	NN	O	O
13C	NN	O	O
)	NN	O	O
-	NN	O	O
galactose	NN	O	O
by	NN	O	O
the	NN	O	O
patient	NN	O	O
homozygous	NN	O	O
for	NN	O	O
S135L	NN	O	O
and	NN	O	O
various	NN	O	O
degrees	NN	O	O
of	NN	O	O
enzyme	NN	O	O
impairment	NN	O	O
among	NN	O	O
different	NN	O	O
tissues	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
The	NN	O	O
S135L	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
GALT	NN	O	O
gene	NN	O	O
is	NN	O	O
a	NN	O	O
prevalent	NN	O	O
cause	NN	O	O
of	NN	O	O
galactosemia	NN	O	B-Disease
among	NN	O	O
black	NN	O	O
patients	NN	O	O
.	NN	O	O

Because	NN	O	O
GALT	NN	O	O
activity	NN	O	O
varies	NN	O	O
in	NN	O	O
different	NN	O	O
tissues	NN	O	O
of	NN	O	O
patients	NN	O	O
homozygous	NN	O	O
for	NN	O	O
S135L	NN	O	O
,	NN	O	O
they	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
better	NN	O	O
clinical	NN	O	O
outcome	NN	O	O
than	NN	O	O
patients	NN	O	O
who	NN	O	O
are	NN	O	O
homozygous	NN	O	O
for	NN	O	O
Q188R	NN	O	O
when	NN	O	O
both	NN	O	O
are	NN	O	O
treated	NN	O	O
from	NN	O	O
infancy	NN	O	O
.	NN	O	O
.	NN	O	O

A	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
20	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
20	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
,	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
which	NN	O	O
show	NN	O	O
positive	NN	O	O
evidence	NN	O	O
of	NN	O	O
linkage	NN	O	O
to	NN	O	O
chromosome	NN	O	O
17q12	NN	O	O
for	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
.	NN	O	O

BRCA1	NN	O	O
mutations	NN	O	O
cosegregating	NN	O	O
with	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
16	NN	O	O
families	NN	O	O
,	NN	O	O
including	NN	O	O
1	NN	O	O
family	NN	O	O
with	NN	O	O
a	NN	O	O
case	NN	O	O
of	NN	O	O
male	NN	O	B-Disease
breast	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Nine	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
mutations	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
generate	NN	O	O
a	NN	O	O
premature	NN	O	O
stop	NN	O	O
codon	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
BRCA1	NN	O	O
protein	NN	O	O
of	NN	O	O
2	NN	O	O
%	NN	O	O
-	NN	O	O
88	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
expected	NN	O	O
normal	NN	O	O
length	NN	O	O
.	NN	O	O

Two	NN	O	O
mutations	NN	O	O
altered	NN	O	O
the	NN	O	O
RING	NN	O	O
finger	NN	O	O
domain	NN	O	O
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
12	NN	O	O
families	NN	O	O
,	NN	O	O
and	NN	O	O
cDNA	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
an	NN	O	O
abnormal	NN	O	O
or	NN	O	O
missing	NN	O	O
BRCA1	NN	O	O
transcript	NN	O	O
in	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
8	NN	O	O
remaining	NN	O	O
families	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
eight	NN	O	O
mutations	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
reduced	NN	O	O
quantity	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
transcript	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
detect	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
4	NN	O	O
of	NN	O	O
the	NN	O	O
20	NN	O	O
families	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
1	NN	O	O
of	NN	O	O
these	NN	O	O
was	NN	O	O
clearly	NN	O	O
linked	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
expected	NN	O	O
that	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
clear	NN	O	O
examples	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	B-Disease
-	NN	O	I-Disease
ovarian	NN	O	I-Disease
syndrome	NN	O	I-Disease
will	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
germ	NN	O	O
-	NN	O	O
line	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O
.	NN	O	O

Brca1	NN	O	B-Disease
deficiency	NN	O	I-Disease
results	NN	O	O
in	NN	O	O
early	NN	O	O
embryonic	NN	O	B-Disease
lethality	NN	O	I-Disease
characterized	NN	O	O
by	NN	O	O
neuroepithelial	NN	O	B-Disease
abnormalities	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
gene	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
zinc	NN	O	O
-	NN	O	O
finger	NN	O	O
protein	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
account	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
families	NN	O	O
with	NN	O	O
both	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
some	NN	O	O
non	NN	O	O
-	NN	O	O
familial	NN	O	O
sporadic	NN	O	O
ovarian	NN	O	B-Disease
cancers	NN	O	I-Disease
.	NN	O	O

The	NN	O	O
loss	NN	O	O
of	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
tumours	NN	O	B-Disease
of	NN	O	O
individuals	NN	O	O
carrying	NN	O	O
one	NN	O	O
nonfunctional	NN	O	O
BRCA1	NN	O	O
allele	NN	O	O
suggests	NN	O	O
that	NN	O	O
BRCA1	NN	O	O
encodes	NN	O	O
a	NN	O	O
tumour	NN	O	B-Disease
suppressor	NN	O	O
that	NN	O	O
may	NN	O	O
inhibit	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
mammary	NN	O	O
epithelial	NN	O	O
cells	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
normal	NN	O	O
tissue	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
generate	NN	O	O
a	NN	O	O
potential	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
cancer	NN	O	B-Disease
susceptibility	NN	O	O
associated	NN	O	O
with	NN	O	O
loss	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
function	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
created	NN	O	O
a	NN	O	O
mouse	NN	O	O
line	NN	O	O
carrying	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
Brca1	NN	O	O
allele	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
mice	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
mutant	NN	O	O
allele	NN	O	O
indicate	NN	O	O
that	NN	O	O
Brca1	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
normal	NN	O	O
development	NN	O	O
,	NN	O	O
as	NN	O	O
these	NN	O	O
mice	NN	O	O
died	NN	O	O
in	NN	O	O
utero	NN	O	O
between	NN	O	O
10	NN	O	O
and	NN	O	O
13	NN	O	O
days	NN	O	O
of	NN	O	O
gestation	NN	O	O
(	NN	O	O
E10	NN	O	O
-	NN	O	O
E13	NN	O	O
)	NN	O	O
.	NN	O	O

Abnormalities	NN	O	O
in	NN	O	O
Brca1	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
embryos	NN	O	O
were	NN	O	O
most	NN	O	O
evident	NN	O	O
in	NN	O	O
the	NN	O	O
neural	NN	O	O
tube	NN	O	O
,	NN	O	O
with	NN	O	O
40	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
embryos	NN	O	O
presenting	NN	O	O
with	NN	O	O
varying	NN	O	O
degrees	NN	O	O
of	NN	O	O
spina	NN	O	B-Disease
bifida	NN	O	I-Disease
and	NN	O	O
anencephaly	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
neuroepithelium	NN	O	O
in	NN	O	O
Brca1	NN	O	B-Disease
-	NN	O	I-Disease
deficient	NN	O	I-Disease
embryos	NN	O	O
appeared	NN	O	O
disorganized	NN	O	O
,	NN	O	O
with	NN	O	O
signs	NN	O	O
of	NN	O	O
both	NN	O	O
rapid	NN	O	O
proliferation	NN	O	O
and	NN	O	O
excessive	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ALD	NN	O	O
-	NN	O	O
gene	NN	O	O
of	NN	O	O
20	NN	O	O
families	NN	O	O
with	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
/	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
.	NN	O	O

Adrenoleukodystrophy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
)	NN	O	O
,	NN	O	O
an	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
inherited	NN	O	I-Disease
metabolic	NN	O	I-Disease
disorder	NN	O	I-Disease
,	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
inborn	NN	O	B-Disease
peroxisomal	NN	O	I-Disease
disease	NN	O	I-Disease
.	NN	O	O

It	NN	O	O
leads	NN	O	O
to	NN	O	O
demyelination	NN	O	B-Disease
in	NN	O	I-Disease
the	NN	O	I-Disease
central	NN	O	I-Disease
and	NN	O	I-Disease
peripheral	NN	O	I-Disease
nervous	NN	O	I-Disease
system	NN	O	I-Disease
.	NN	O	O

Defective	NN	O	O
beta	NN	O	O
-	NN	O	O
oxidation	NN	O	O
of	NN	O	O
saturated	NN	O	O
very	NN	O	O
long	NN	O	O
chain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
(	NN	O	O
VLCFAs	NN	O	O
;	NN	O	O
C22	NN	O	O
0	NN	O	O
-	NN	O	O
C26	NN	O	O
0	NN	O	O
)	NN	O	O
in	NN	O	O
peroxisomes	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
lead	NN	O	O
to	NN	O	O
an	NN	O	O
accumulation	NN	O	O
of	NN	O	O
VLCFAs	NN	O	O
in	NN	O	O
leukoid	NN	O	O
areas	NN	O	O
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
peripheral	NN	O	O
nerves	NN	O	O
,	NN	O	O
adrenal	NN	O	O
gland	NN	O	O
,	NN	O	O
and	NN	O	O
blood	NN	O	O
.	NN	O	O

The	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
identified	NN	O	O
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
745	NN	O	O
-	NN	O	O
amino	NN	O	O
-	NN	O	O
acid	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
screened	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenoleukodystrophy	NN	O	B-Disease
/	NN	O	O
adrenomyeloneuropathy	NN	O	B-Disease
(	NN	O	O
ALD	NN	O	B-Disease
/	NN	O	O
AMN	NN	O	B-Disease
)	NN	O	O
from	NN	O	O
20	NN	O	O
kindreds	NN	O	O
for	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
.	NN	O	O

Eleven	NN	O	O
missense	NN	O	O
and	NN	O	O
two	NN	O	O
nonsense	NN	O	O
mutations	NN	O	O
,	NN	O	O
five	NN	O	O
deletions	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
insertion	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
eight	NN	O	O
reverse	NN	O	O
transcribed	NN	O	O
fragments	NN	O	O
of	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
-	NN	O	O
gene	NN	O	O
mRNA	NN	O	O
.	NN	O	O

Four	NN	O	O
mutations	NN	O	O
could	NN	O	O
be	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
de	NN	O	O
novo	NN	O	O
.	NN	O	O

All	NN	O	O
mutations	NN	O	O
could	NN	O	O
be	NN	O	O
confirmed	NN	O	O
in	NN	O	O
carriers	NN	O	O
by	NN	O	O
sequencing	NN	O	O
genomic	NN	O	O
DNA	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
mutation	NN	O	O
and	NN	O	O
the	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
phenotype	NN	O	O
could	NN	O	O
be	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
ALD	NN	O	B-Disease
gene	NN	O	O
of	NN	O	O
30	NN	O	O
healthy	NN	O	O
persons	NN	O	O
.	NN	O	O
.	NN	O	O

The	NN	O	O
murine	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
gene	NN	O	O
Brca1	NN	O	O
maps	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
11D	NN	O	O
.	NN	O	O

The	NN	O	O
recently	NN	O	O
cloned	NN	O	O
human	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
susceptibility	NN	O	O
gene	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
is	NN	O	O
located	NN	O	O
on	NN	O	O
human	NN	O	O
chromosome	NN	O	O
17q21	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
murine	NN	O	O
genomic	NN	O	O
clones	NN	O	O
containing	NN	O	O
Brca1	NN	O	O
as	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
in	NN	O	O
generating	NN	O	O
a	NN	O	O
mouse	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
function	NN	O	O
.	NN	O	O

A	NN	O	O
mouse	NN	O	O
genomic	NN	O	O
library	NN	O	O
was	NN	O	O
screened	NN	O	O
using	NN	O	O
probes	NN	O	O
corresponding	NN	O	O
to	NN	O	O
exon	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
.	NN	O	O

Two	NN	O	O
overlapping	NN	O	O
mouse	NN	O	O
clones	NN	O	O
were	NN	O	O
identified	NN	O	O
that	NN	O	O
hybridized	NN	O	O
to	NN	O	O
human	NN	O	O
BRCA1	NN	O	O
exons	NN	O	O
9	NN	O	O
-	NN	O	O
12	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
1	NN	O	O
.	NN	O	O

4	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
these	NN	O	O
clones	NN	O	O
corresponding	NN	O	O
to	NN	O	O
part	NN	O	O
of	NN	O	O
human	NN	O	O
exon	NN	O	O
11	NN	O	O
revealed	NN	O	O
72	NN	O	O
%	NN	O	O
nucleic	NN	O	O
acid	NN	O	O
identity	NN	O	O
but	NN	O	O
only	NN	O	O
50	NN	O	O
%	NN	O	O
amino	NN	O	O
acid	NN	O	O
identity	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
longest	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
Brca1	NN	O	O
genomic	NN	O	O
clones	NN	O	O
maps	NN	O	O
to	NN	O	O
chromosome	NN	O	O
11D	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
two	NN	O	O
-	NN	O	O
color	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

The	NN	O	O
synteny	NN	O	O
to	NN	O	O
human	NN	O	O
chromosome	NN	O	O
17	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
cohybridization	NN	O	O
with	NN	O	O
the	NN	O	O
mouse	NN	O	O
probe	NN	O	O
for	NN	O	O
the	NN	O	O
NF1	NN	O	O
-	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
comparative	NN	O	O
study	NN	O	O
confirms	NN	O	O
that	NN	O	O
the	NN	O	O
relative	NN	O	O
location	NN	O	O
of	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
conserved	NN	O	O
between	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

Atelosteogenesis	NN	O	B-Disease
type	NN	O	I-Disease
II	NN	O	I-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
sulfate	NN	O	O
-	NN	O	O
transporter	NN	O	O
gene	NN	O	O
(	NN	O	O
DTDST	NN	O	O
)	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
phenotypic	NN	O	O
series	NN	O	O
involving	NN	O	O
three	NN	O	O
chondrodysplasias	NN	O	B-Disease
.	NN	O	O

Atelosteogenesis	NN	O	B-Disease
type	NN	O	I-Disease
II	NN	O	I-Disease
(	NN	O	O
AO	NN	O	B-Disease
II	NN	O	I-Disease
)	NN	O	O
is	NN	O	O
a	NN	O	O
neonatally	NN	O	O
lethal	NN	O	O
chondrodysplasia	NN	O	B-Disease
whose	NN	O	O
clinical	NN	O	O
and	NN	O	O
histological	NN	O	O
characteristics	NN	O	O
resemble	NN	O	O
those	NN	O	O
of	NN	O	O
another	NN	O	O
chondrodysplasia	NN	O	B-Disease
,	NN	O	O
the	NN	O	O
much	NN	O	O
less	NN	O	O
severe	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
(	NN	O	O
DTD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

The	NN	O	O
similarity	NN	O	O
suggests	NN	O	O
a	NN	O	O
shared	NN	O	O
pathogenesis	NN	O	O
involving	NN	O	O
lesions	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
biochemical	NN	O	O
pathway	NN	O	O
and	NN	O	O
perhaps	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
.	NN	O	O

DTD	NN	O	B-Disease
is	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
recently	NN	O	O
identified	NN	O	O
diastrophic	NN	O	B-Disease
dysplasia	NN	O	I-Disease
sulfate	NN	O	O
-	NN	O	O
transporter	NN	O	O
gene	NN	O	O
(	NN	O	O
DTDST	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
AOII	NN	O	B-Disease
patients	NN	O	O
also	NN	O	O
have	NN	O	O
DTDST	NN	O	O
mutations	NN	O	O
,	NN	O	O
which	NN	O	O
lead	NN	O	O
to	NN	O	O
defective	NN	O	O
uptake	NN	O	O
of	NN	O	O
inorganic	NN	O	O
sulfate	NN	O	O
and	NN	O	O
insufficient	NN	O	O
sulfation	NN	O	O
of	NN	O	O
macromolecules	NN	O	O
by	NN	O	O
patient	NN	O	O
mesenchymal	NN	O	O
cells	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Together	NN	O	O
with	NN	O	O
our	NN	O	O
recent	NN	O	O
observation	NN	O	O
that	NN	O	O
a	NN	O	O
third	NN	O	O
even	NN	O	O
more	NN	O	O
severe	NN	O	O
chondrodysplasia	NN	O	B-Disease
,	NN	O	O
achondrogenesis	NN	O	O
type	NN	O	O
IB	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
DTDST	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
phenotypic	NN	O	O
series	NN	O	O
of	NN	O	O
three	NN	O	O
chondrodysplasias	NN	O	B-Disease
of	NN	O	O
increasing	NN	O	O
severity	NN	O	O
caused	NN	O	O
by	NN	O	O
lesions	NN	O	O
in	NN	O	O
a	NN	O	O
single	NN	O	O
sulfate	NN	O	O
-	NN	O	O
transporter	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
severity	NN	O	O
of	NN	O	O
the	NN	O	O
phenotype	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
predicted	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
mutations	NN	O	O
on	NN	O	O
the	NN	O	O
residual	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
DTDST	NN	O	O
protein	NN	O	O
.	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
and	NN	O	O
phenotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
six	NN	O	O
recurrent	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
61	NN	O	O
families	NN	O	O
:	NN	O	O
results	NN	O	O
of	NN	O	O
an	NN	O	O
international	NN	O	O
study	NN	O	O
.	NN	O	O

Several	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
geographically	NN	O	O
diverse	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
mutation	NN	O	O
origin	NN	O	O
and	NN	O	O
mutation	NN	O	O
-	NN	O	O
specific	NN	O	O
phenotypes	NN	O	O
due	NN	O	O
to	NN	O	O
BRCA1	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
haplotype	NN	O	O
of	NN	O	O
nine	NN	O	O
polymorphic	NN	O	O
markers	NN	O	O
within	NN	O	O
or	NN	O	O
immediately	NN	O	O
flanking	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
locus	NN	O	O
in	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
61	NN	O	O
breast	NN	O	B-Disease
/	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
families	NN	O	O
selected	NN	O	O
for	NN	O	O
having	NN	O	O
one	NN	O	O
of	NN	O	O
six	NN	O	O
recurrent	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

Tests	NN	O	O
of	NN	O	O
both	NN	O	O
mutations	NN	O	O
and	NN	O	O
family	NN	O	O
-	NN	O	O
specific	NN	O	O
differences	NN	O	O
in	NN	O	O
age	NN	O	O
at	NN	O	O
diagnosis	NN	O	O
were	NN	O	O
not	NN	O	O
significant	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
six	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
relative	NN	O	O
proportions	NN	O	O
of	NN	O	O
cases	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
was	NN	O	O
suggestive	NN	O	O
of	NN	O	O
an	NN	O	O
effect	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.	NN	O	O
069	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
57	NN	O	O
%	NN	O	O
of	NN	O	O
women	NN	O	O
presumed	NN	O	O
affected	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
1294	NN	O	O
del	NN	O	O
40	NN	O	O
BRCA1	NN	O	O
mutation	NN	O	O
having	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
,	NN	O	O
compared	NN	O	O
with	NN	O	O
14	NN	O	O
%	NN	O	O
of	NN	O	O
affected	NN	O	O
women	NN	O	O
with	NN	O	O
the	NN	O	O
splice	NN	O	O
-	NN	O	O
site	NN	O	O
mutation	NN	O	O
in	NN	O	O
intron	NN	O	O
5	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

For	NN	O	O
the	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
studied	NN	O	O
here	NN	O	O
,	NN	O	O
the	NN	O	O
individual	NN	O	O
mutations	NN	O	O
are	NN	O	O
estimated	NN	O	O
to	NN	O	O
have	NN	O	O
arisen	NN	O	O
9	NN	O	O
-	NN	O	O
170	NN	O	O
generations	NN	O	O
ago	NN	O	O
.	NN	O	O

In	NN	O	O
general	NN	O	O
,	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
haplotype	NN	O	O
conservation	NN	O	O
across	NN	O	O
the	NN	O	O
region	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
with	NN	O	O
haplotype	NN	O	O
differences	NN	O	O
most	NN	O	O
often	NN	O	O
due	NN	O	O
to	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
short	NN	O	O
-	NN	O	O
tandem	NN	O	O
-	NN	O	O
repeat	NN	O	O
markers	NN	O	O
,	NN	O	O
although	NN	O	O
some	NN	O	O
likely	NN	O	O
instances	NN	O	O
of	NN	O	O
recombination	NN	O	O
also	NN	O	O
were	NN	O	O
observed	NN	O	O
.	NN	O	O

For	NN	O	O
several	NN	O	O
of	NN	O	O
the	NN	O	O
instances	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
evidence	NN	O	O
for	NN	O	O
multiple	NN	O	O
,	NN	O	O
independent	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
mutational	NN	O	O
events	NN	O	O
.	NN	O	O

Isolation	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
and	NN	O	O
genetic	NN	O	O
mapping	NN	O	O
to	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
11	NN	O	O
.	NN	O	O

The	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
is	NN	O	O
in	NN	O	O
large	NN	O	O
part	NN	O	O
responsible	NN	O	O
for	NN	O	O
hereditary	NN	O	B-Disease
human	NN	O	I-Disease
breast	NN	O	I-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
Brca1	NN	O	O
homologue	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
genomic	NN	O	O
P1	NN	O	O
clones	NN	O	O
that	NN	O	O
contain	NN	O	O
most	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
all	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
Brca1	NN	O	O
locus	NN	O	O
.	NN	O	O

DNA	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
coding	NN	O	O
regions	NN	O	O
are	NN	O	O
75	NN	O	O
%	NN	O	O
identical	NN	O	O
at	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
level	NN	O	O
while	NN	O	O
the	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
identity	NN	O	O
is	NN	O	O
only	NN	O	O
58	NN	O	O
%	NN	O	O
.	NN	O	O

A	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
variant	NN	O	O
in	NN	O	O
the	NN	O	O
Brca1	NN	O	O
locus	NN	O	O
was	NN	O	O
identified	NN	O	O
and	NN	O	O
used	NN	O	O
to	NN	O	O
map	NN	O	O
this	NN	O	O
gene	NN	O	O
on	NN	O	O
a	NN	O	O
(	NN	O	O
Mus	NN	O	O
m	NN	O	O
.	NN	O	O
musculus	NN	O	O
Czech	NN	O	O
II	NN	O	O
x	NN	O	O
C57BL	NN	O	O
/	NN	O	O
KsJ	NN	O	O
)	NN	O	O
F1	NN	O	O
x	NN	O	O
C57BL	NN	O	O
/	NN	O	O
KsJ	NN	O	O
intersubspecific	NN	O	O
backcross	NN	O	O
to	NN	O	O
distal	NN	O	O
mouse	NN	O	O
chromosome	NN	O	O
11	NN	O	O
.	NN	O	O

The	NN	O	O
mapping	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
highly	NN	O	O
syntenic	NN	O	O
with	NN	O	O
human	NN	O	O
chromosome	NN	O	O
17	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
Southern	NN	O	O
and	NN	O	O
Northern	NN	O	O
analyses	NN	O	O
,	NN	O	O
confirms	NN	O	O
that	NN	O	O
we	NN	O	O
isolated	NN	O	O
the	NN	O	O
murine	NN	O	O
Brca1	NN	O	O
homologue	NN	O	O
rather	NN	O	O
than	NN	O	O
a	NN	O	O
related	NN	O	O
RING	NN	O	O
finger	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
Brca1	NN	O	O
homologue	NN	O	O
will	NN	O	O
facilitate	NN	O	O
the	NN	O	O
creation	NN	O	O
of	NN	O	O
mouse	NN	O	O
models	NN	O	O
for	NN	O	O
germline	NN	O	O
BRCA1	NN	O	B-Disease
defects	NN	O	I-Disease
.	NN	O	O
.	NN	O	O

Emerin	NN	O	B-Disease
deficiency	NN	O	I-Disease
at	NN	O	O
the	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Emery	NN	O	B-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
STA	NN	O	O
gene	NN	O	O
at	NN	O	O
the	NN	O	O
Xq28	NN	O	O
locus	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
X	NN	O	B-Disease
-	NN	O	I-Disease
linked	NN	O	I-Disease
Emery	NN	O	I-Disease
-	NN	O	I-Disease
Dreifuss	NN	O	I-Disease
muscular	NN	O	I-Disease
dystrophy	NN	O	I-Disease
(	NN	O	O
EDMD	NN	O	B-Disease
)	NN	O	O
.	NN	O	O

This	NN	O	O
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
hitherto	NN	O	O
unknown	NN	O	O
protein	NN	O	O
named	NN	O	O
emerin	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
emerin	NN	O	O
,	NN	O	O
we	NN	O	O
raised	NN	O	O
two	NN	O	O
antisera	NN	O	O
against	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
fragments	NN	O	O
predicted	NN	O	O
from	NN	O	O
emerin	NN	O	O
cDNA	NN	O	O
.	NN	O	O

Using	NN	O	O
both	NN	O	O
antisera	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
positive	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
staining	NN	O	O
in	NN	O	O
skeletal	NN	O	O
,	NN	O	O
cardiac	NN	O	O
and	NN	O	O
smooth	NN	O	O
muscles	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
controls	NN	O	O
and	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
neuromuscular	NN	O	B-Disease
diseases	NN	O	I-Disease
other	NN	O	O
than	NN	O	O
EDMD	NN	O	B-Disease
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
deficiency	NN	O	O
in	NN	O	O
immunofluorescent	NN	O	O
staining	NN	O	O
of	NN	O	O
skeletal	NN	O	O
and	NN	O	O
cardiac	NN	O	O
muscle	NN	O	O
from	NN	O	O
EDMD	NN	O	B-Disease
patients	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

A	NN	O	O
34	NN	O	O
kD	NN	O	O
protein	NN	O	O
is	NN	O	O
immunoreactive	NN	O	O
with	NN	O	O
the	NN	O	O
antisera	NN	O	O
-	NN	O	O
-	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
equivalent	NN	O	O
to	NN	O	O
that	NN	O	O
predicted	NN	O	O
for	NN	O	O
emerin	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
the	NN	O	O
specific	NN	O	O
deficiency	NN	O	B-Disease
of	NN	O	I-Disease
emerin	NN	O	I-Disease
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
of	NN	O	O
muscle	NN	O	O
cells	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
EDMD	NN	O	B-Disease
.	NN	O	O
.	NN	O	O

Growth	NN	O	B-Disease
retardation	NN	O	I-Disease
and	NN	O	O
tumour	NN	O	B-Disease
inhibition	NN	O	O
by	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Inherited	NN	O	O
mutations	NN	O	O
in	NN	O	O
BRCA1	NN	O	O
predispose	NN	O	O
to	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
,	NN	O	O
but	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
BRCA1	NN	O	O
in	NN	O	O
sporadic	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
has	NN	O	O
previously	NN	O	O
been	NN	O	O
elusive	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
retroviral	NN	O	O
transfer	NN	O	O
of	NN	O	O
the	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
BRCA1	NN	O	O
gene	NN	O	O
inhibits	NN	O	O
growth	NN	O	O
in	NN	O	O
vitro	NN	O	O
of	NN	O	O
all	NN	O	O
breast	NN	O	B-Disease
and	NN	O	I-Disease
ovarian	NN	O	I-Disease
cancer	NN	O	I-Disease
cell	NN	O	O
lines	NN	O	O
tested	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
colon	NN	O	B-Disease
or	NN	O	I-Disease
lung	NN	O	I-Disease
cancer	NN	O	I-Disease
cells	NN	O	O
or	NN	O	O
fibroblasts	NN	O	O
.	NN	O	O

Mutant	NN	O	O
BRCA1	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
growth	NN	O	O
of	NN	O	O
breast	NN	O	B-Disease
cancer	NN	O	I-Disease
cells	NN	O	O
;	NN	O	O
ovarian	NN	O	B-Disease
cancer	NN	O	I-Disease
cell	NN	O	O
growth	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
3	NN	O	O
BRCA1	NN	O	O
mutations	NN	O	O
.	NN	O	O

Development	NN	O	O
of	NN	O	O
MCF	NN	O	B-Disease
-	NN	O	I-Disease
7	NN	O	I-Disease
tumours	NN	O	I-Disease
in	NN	O	O
nude	NN	O	O
mice	NN	O	O
is	NN	O	O
inhibited	NN	O	O
when	NN	O	O
MCF	NN	O	O
-	NN	O	O
7	NN	O	O
cells	NN	O	O
are	NN	O	O
transfected	NN	O	O
with	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mutant	NN	O	O
,	NN	O	O
BRCA1	NN	O	O
.	NN	O	O

Most	NN	O	O
importantly	NN	O	O
,	NN	O	O
among	NN	O	O
mice	NN	O	O
with	NN	O	O
established	NN	O	O
MCF	NN	O	B-Disease
-	NN	O	I-Disease
7	NN	O	I-Disease
tumours	NN	O	I-Disease
,	NN	O	O
peritoneal	NN	O	O
treatment	NN	O	O
with	NN	O	O
a	NN	O	O
retroviral	NN	O	O
vector	NN	O	O
expressing	NN	O	O
wild	NN	O	O
-	NN	O	O
type	NN	O	O
BRCA1	NN	O	O
significantly	NN	O	O
inhibits	NN	O	O
tumour	NN	O	B-Disease
growth	NN	O	O
and	NN	O	O
increased	NN	O	O
survival	NN	O	O
.	NN	O	O
.	NN	O	O

