Medical Research Society
89
A GENERAL ABNORMALITY O F BONE M A T R I X I N
OSTEOWROSIS
B. F E R R I S ~ ,R~DODDS',
AND J. CHAYEN
L. KLENERMAN~, L. B1TEN.d
'Division of C l l u l a r Biology, Kennedy I n s t i t u t e ,
London W6 and 'Department o f Orthopaedics,
Northwick Park H o s p i t a l , Harrow.
I n p a t i e n t s w i t h o s t e o p o r o s i s t h e bone i s
g e n e r a l l y h e l d to be reduced i n q u a n t i t y , b u t
normal i n q u a l i t y . However r e c e n t work (Kent
e t a l , J. Bone Jt. Surg., 658, 189,1983) showed
t h a t a t t h e site of subcapital fractures, t h e
o r g a n i s a t i o n o f t h e glycosaminoglycans (GAGS)
w a s markedly d e c r e a s e d , w i t h no change i n t o t a l
c o n t e n t . To t e s t whether t h i s p e r t a i n e d o n l y t o
the f r a c t u r e site, or w a s a generalised f e a t u r e
of o s t e o p o r o s i s , w e have examined i l i a c crest
b i o p s i e s from p a t i e n t s w i t h o s t e o p o r o t i c proximal
femoral f r a c t u r e s (10 s u b c a p i t a l and 10 t r o c h a n t eric) and from normal s u b j e c t s having hone g r a f t s
( s i x ) or post-mortem(l0).
The o r i e n t a t i o n of c o l l a g e n w a s examined d i r e c t l y
by q u a n t i t a t i v e p o l a r i s e d l i g h t microscopy i n
unfixed, undemineralised c r y o s t a t s e c t i o n s (lourn).
The t o t a l amount of GAGS and r e l a t e d m o i e t i e s
was measured by microdensitometry a f t e r s t a i n i n g
such s e c t i o n s w i t h Alcian b l u e i n t h e p r e s e n c e
o f e i t h e r 0.025M o r 0.5M MgC12. The o r i e n t a t i o n
of t h e GAGS and related molecules w a s measured
by t h e b i r e f r i n g e n c e induced by such s t a i n i n g .
Collagen o r i e n t a t i o n , as measured by i t s
b i r e f r i n g e n c e ( o p t i c a l p a t h d i f f e r e n c e : 0.p.d.
i n n m ) , i n a l l s e c t i o n s w a s c o n s t a n t (16.4 2
0.3nm; mean 5 SEM; n = 36), a s w a s t h e t o t a l
c o n t e n t o f GAGS and r e l a t e d m o i e t i e s . However
t h e o r i e n t a t i o n of GAGS and r e l a t e d m o i e t i e s ,
a s a s s e s s e d by t h e induced b i r e f r i n g e n c e ( w i t h
0.5M MgC12), i n t h e o s t e o p o r o t i c specimens w a s
o n l y 65% of that found i n t h e c o n t r o l s (10.4 2
0.4 a s a g a i n s t 16.0 5 0.8nm; p<O.OOl). These
1,
v a l u e s are s i m i l a r i n t r e n d t o t h o s e found i n
comparable bone from t h e femoral neck (7.8 2 0.2
81s a g a i n s t 17.3 2 0.7nm).
Consequently it a p p e a r s t h a t t h e abnormal
o r i e n t a t i o n of GAGS and r e l a t e d m o i e t i e s i n
o s t e o p o r o t i c f r a c t u r e s i s n o t confined t o t h e
f r a c t u r e - s i t e b u t may, t o a lesser d e g r e e , b e a
g e n e r a l i s e d a l t e r a t i o n i n t h e q u a l i t y o f bone i n
osteoporosis.
--
90
INTERFERON MODIFIES
HEPATITIS B VIRUS
PROTEIN EXPRESSION: IDENTIFICATION OF AN
INTERFERON SENSITIVE NUCLEOTIDE SEQUENCE I N THE
VIRUS GENOME
A.M.L.
LEVER, M. PIGNATELLI and H.C.
THOMAS
Academic Department of Medicine, Royal F r e e
H o s p i t a l , Hampstead, London, England, UK
Chronic h e p a t i t i s B v i r u s (HBV) i n f e c t i o n
a f f e c t s 200 m i l l i o n people worldwide. I n t e r f e r on(1FN) h a s been used t h e r a p e u t i c a l l y i n t h i s
d i s e a s e w i t h seroconversion r a t e s of about 50%
i n r e c e n t s t u d i e s . D e s t r u c t i o n of HBV i n f e c t e d
c e l l s .appears t o depend on r e c o g n i t i o n of t h e
HB c o r e / e a n t i g e n s on t h e hepatocyte membrane
by c y t o t o x i c T lymphocytes. The v i r u s DNA i s a
p a r t i a l l y double s t r a n d e d molecule which
a s h o r t transproduces two mRNA t r a n s c r i p t s :
33 P
c r i p t coding only f o r t h e s u r f a c e a n t i g e n (HBsAd
and a long t r a n s c r i p t coding f o r several p r o t e i r s
i n c l u d i n g t h e HB c o r e a n t i g e n (HBcAg).
L i v e r biopsy specimens from p a t i e n t s w i t h
u n t r e a t e d c h r o n i c HEY i n f e c t i o n and from p a t i e n t s
i n whom I F N therapy had begun, were examined by
immunofluoresence f o r e x p r e s s i o n of HBsAg and
HBcAg p r o t e i n s .
I n t e r f e r o n t r e a t m e n t produced
a s e l e c t i v e i n c r e a s e i n HBc and a d e c r e a s e i n
HBs e x p r e s s i o n i n v i r u s i n f e c t e d cells.
We sought evidence f o r IFN s e n s i t i v e DNA
sequences i n t h e v i r u s genome and found a
n u c l e o t i d e sequence homologous w i t h a consensus
sequence upstream f o r several IFN induced
p r o t e i n s i n t h e human genome. This sequence
occurs a t t h e s t a r t of t h e long mRNA t r a n s c r i p t
It i s p r o b a b l e t h a t IFN is a c t i n g
i n the virus.
a t t h i s s i t e and d i r e c t l y i n c r e a s i n g HBcAg gene
t r a n s c r i p t i o n and e x p r e s s i o n . T h i s may be one
mechanism by which ITN a c t s t h e r a p e u t i c a l l y i n
these patients.
I n c h r o n i c HBV i n f e c t i o n t h e v i r u s i n t e g r a t e s
i n t o t h e h o s t DNA i n a p r o p o r t i o n of h e p a t o c y t e s
The DNA homology between v i r u s and h o s t occurs
a t t h e s i t e on t h e v i r a l DNA known t o be importa n t f o r HBV i n t e g r a t i o n i n t o t h e h o s t genome and
t h e v i r u s may t h e r e f o r e p r e f e r e n t i a l l y i n t e g r a t e
a t h o s t IFN s e n s i t i v e s i t e s . This mechanism
could l e a d t o both HBV being t r a n s c r i b e d when
IFN a c t s on t h e c e l l and a l s o might i n t e r f e r e
w i t h t h e p r o d u c t i o n of a n t i v i r a l p r o t e i n s by t h e
c e l l i n response t o IFN.
91
CORONARY ANGIOGRAPHIC FINDINGS I N INFARCT
RELATED VESSELS BEFORE AND AFTER INFARCTION
D. HACKETT, J. VERWILGHEN, G. DAVIES,
S. CHIERCHIA, A. MASER1
D i v i s i o n of C a r d i o v a s c u l a r Disease, Royal
P o s t g r a d u a t e Medical School, Hammersmith
H o s p i t a l , London, W.12
The
possibility
of
identifying
coronary
a r t e r i o g r a p h i c f e a t u r e s p r e d i c t i v e of f u t u r e
o c c l u s i o n and myocardial i n f a r c t i o n (MI) would
have
important
therapeutic
and
prognostic
i m p l i c a t i o n s . From 1978-85 w e i d e n t i f i e d 10
patients
(pts)
who had
e l e c t i v e coronary
a r t e r i o g r a p h y f o r c h r o n i c s t a b l e angina and
s u b s e q u e n t l y developed a n M I .
I n a l l , a preand post-MI a r t e r i o g r a m had been o b t a i n e d a t a
remote time from M I a n d ' n o n e had undergone any
surgical o r invasive therapeutic interventions.
The
pre-MI
arteriogram
showed
an already
s t e n o s e d i n f a r c t r e l a t e d v e s s e l t o be p r e s e n t
i n 5 p t s (group 1); none had
identifiable
changes of s t e n o s i s s e v e r i t y i n t h e p o s t M I
s t u d y (from 60+21.5% t o 63+26.4%; mean
SD;
p=NS). The othTr 5 p t s (gFoup 2 ) had normal
pre-MI a r t e r i o g r a m s ( 3 p t s ) o r only minor
irregularities,
but showed marked l o c a l i s e d
p r o g r e s s i o n of coronary o b s t r u c t i o n i n t h e
i n f a r c t r e l a t e d v e s s e l i n a l l t h e post-MI
studi.es (from 627.1% t o 8628.9%; p 0.01). S i t e
and s i z e of M I was n o t p r e d i c t a b l e by pre-MI
arteriographic
appearances,
severity
of
stenosis,
the
presence
or
absence
of
c o l l a t e r a l s , o r t i m e e l a p s e d between angiograms.
D i f f e r e n c e s i n p r o g r e s s i o n of s t e n o s e s between
groups 1 and 2 a l s o could n o t be r e l a t e d t o t h e
presence of coronary r i s k f a c t o r s , previous M I ,
time e l a p s e d between angiograms, d u r a t i o n from
f i r s t angiogram t o M I , M I s i z e , o r t h e presence
o r absence of c o l l a t e r a l s b e f o r e o r a f t e r M I .
+
Therefore M I can develop i n a r e a s d i s t a l t o
coronary a r t e r i e s w i t h a very v a r i a b l e s e v e r i t y