HISTOLOGICAL AND HISTOCHEMICAL CHANGES OF THE
SKELETAL MUSCLE IN HUMAN CHRONIC
CHAGAS' DISEASE
ANALIA TARATUTO Μ . A. PAGNO TERESA FUMO OLG P. SANZ
R. E. P. S I C A
In a recent electrophysiological investigation Pagano et al. demonstrated
the presence of mild denervation in the skeletal muscle of patients with chronic
Chagas' disease. Later on, the Ramos Mejia Hospital's group confirmed the
findings of Pagano et al.
4and pointed out the spinal alpha motoneurone soma
as the site of the lesion within the motor unit (m.u.)
e.
The aim of the present study has been to obtain information concerning
the eventual histological changes of the skeletal muscle of patients with chronic
Chagas' disease.
M A T E R I A L A N D M E T H O D S
O p e n b i o p s i e s w e r e c a r r i e d o u t u n d e r local a n a e s t h e s i a o n t h e g a s t r o c n e m i u s m e d i a l h e a d m u s c l e i n 7 s u b j e c t s , 5 m a l e s a n d 2 f e m a l e s , a g e d b e t w e e n 25 a n d 44 y e a r s . A t h o r o u g h h i s t o r y w a s o b t a i n e d f r o m e a c h p a t i e n t a n d a c o m p l e t e c l i n i c a l a n d l a b o r a t o r y e x a m i n a t i o n w a s a l s o p e r f o r m e d ; o n t h e s e b a s e s t h e s e l e c t e d s u b j e c t s w e r e j u d g e d t o b e h e a l t h y , b e i n g t h e o n l y a b n o r m a l i t y d e t e c t e d t h e p r e s e n c e of 2 o u t of 3 p o s i t i v e s e r u m t e s t s for Chagas* d i s e a s e . N o n e of t h e m h a d b e e n p r e v i o u s l y t r e a t e d w i t h s p e c i f i c d r u g s .
C o i n c i d e n t a l c a u s e s of m u s c l e d e n e r v a t i o n w e r e a v o i d e d b y r e j e c t i n g p a t i e n t s o v e r t h e a g e o f 60 y e a r s ι ,β a n d t h o s e w i t h a h i s t o r y of t o x i c o r m e t a b o l i c d i s o r d e r s k n o w n to b e a s s o c i a t e d w i t h m u s c l e d e n e r v a t i o n . A l s o w e r e e l i m i n a t e d t h o s e p a t i e n t s w i t h p e r s o n a l o r f a m i l y h i s t o r y of m u s c l e d i s e a s e .
F r e s h m u s c l e s p e c i m e n s w e r e f r o z e n i n i s o p e n t a n e c o o l e d t o -150C<> i n l i q u i d n i t r o g e n . C r y o s t a t s e c t i o n s w e r e c u t a t t h i c k n e s s o f 2 t o 10 urn a n d w e r e s t a i n e d o r r e a c t e d w i t h t h e f o l l o w i n g t e c h n i q u e s : h e m a e t o x i l i n - e o s i n , p e r i o d i c a c i d Schiff, m o d i f i e d Gomori t r i c h r o m e , r e d u c e d n i c o t i n a m i d e a d e n i n e d i n u c l e o t i d e d e h i d r o g e n a s e ( N A D H ) , m i o f i b r i l l a r a d e n o s i n e t r i p h o s p h a t a s e ( A T P a s e ) a t p H 9.4 a n d 4.6. a l k a l i n e p h o s p h a t a s e a n d n o n - s p e c i f i c e s t e r a s e .
R E S U L T S
T h e m o s t c o m m o n f i n d i n g w a s t h e p r e s e n c e o f c l u s t e r s of u n i f o r m t y p e of m u s c l e f i b r e s , e i t h e r t y p e I o r t y p e I I , i n 6 of t h e 7 s t u d i e d p a t i e n t s . B o t h t y p e s of c l u s t e r s , p l a c e d s i d e b y s i d e , c o u l d o f t e n b e f o u n d i n t h e s a m e s p e c i m e n ( F i g u r e 1. T a b l e 1 ) .
S e c t i o n of Clinical E l e c t r o n e u r o p h y s i o l o g y , H o s p i t a l R a m o s M e j i a , B u e n o s A i r e s . A r g e n t i n a .
M A T E R I A L A N D M E T H O D S
O p e n b i o p s i e s w e r e c a r r i e d o u t u n d e r local a n a e s t h e s i a o n t h e g a s t r o c n e m i u s m e d i a l h e a d m u s c l e i n 7 s u b j e c t s , 5 m a l e s a n d 2 f e m a l e s , a g e d b e t w e e n 25 a n d 44 y e a r s . A t h o r o u g h h i s t o r y w a s o b t a i n e d f r o m e a c h p a t i e n t a n d a c o m p l e t e c l i n i c a l a n d l a b o r a t o r y e x a m i n a t i o n w a s a l s o p e r f o r m e d ; o n t h e s e b a s e s t h e s e l e c t e d s u b j e c t s w e r e j u d g e d t o b e h e a l t h y , b e i n g t h e o n l y a b n o r m a l i t y d e t e c t e d t h e p r e s e n c e of 2 o u t of 3 p o s i t i v e s e r u m t e s t s for Chagas* d i s e a s e . N o n e of t h e m h a d b e e n p r e v i o u s l y t r e a t e d w i t h s p e c i f i c d r u g s .
C o i n c i d e n t a l c a u s e s of m u s c l e d e n e r v a t i o n w e r e a v o i d e d b y r e j e c t i n g p a t i e n t s o v e r t h e a g e o f 60 y e a r s ι ,β a n d t h o s e w i t h a h i s t o r y of t o x i c o r m e t a b o l i c d i s o r d e r s k n o w n to b e a s s o c i a t e d w i t h m u s c l e d e n e r v a t i o n . A l s o w e r e e l i m i n a t e d t h o s e p a t i e n t s w i t h p e r s o n a l o r f a m i l y h i s t o r y of m u s c l e d i s e a s e .
F r e s h m u s c l e s p e c i m e n s w e r e f r o z e n i n i s o p e n t a n e c o o l e d t o -150C<> i n l i q u i d n i t r o g e n . C r y o s t a t s e c t i o n s w e r e c u t a t t h i c k n e s s o f 2 t o 10 urn a n d w e r e s t a i n e d o r r e a c t e d w i t h t h e f o l l o w i n g t e c h n i q u e s : h e m a e t o x i l i n - e o s i n , p e r i o d i c a c i d Schiff, m o d i f i e d G o m o r i t r i c h r o m e , r e d u c e d n i c o t i n a m i d e a d e n i n e d i n u c l e o t i d e d e h i d r o g e n a s e ( N A D H ) , m i o f i b r i l l a r a d e n o s i n e t r i p h o s p h a t a s e ( A T P a s e ) a t p H 9.4 a n d 4.6. a l k a l i n e p h o s p h a t a s e a n d n o n - s p e c i f i c e s t e r a s e .
R E S U L T S
328 ARQ- NEURO-PSIQUIATRIA (8Ã0 PAULO) VOL. 36Λ No k, DEZEMBRO, 1978
T h e c l u s t e r s w e r e c o m p o s e d of a b o u t 16 t o 30 f i b r e s , b e i n g s m a l l i n 4 p a t i e n t s w h i l e i n t h e r e m a i n i n g o n e m u c h l a r g e r c l u s t e r s c o u l d b e s e e n ( T a b l e 1 ) .
I n p a t i e n t s V. Ch a n d O. R. s m a l l a n g u l a r f i b r e s , w h i c h s t a i n e d o r r e a c t e d s t r o n g l y w i t h n o n - s p e c i f i c s t e r a s e a n d N A D H t e c h n i q u e s , w e r e a l s o o b s e r v e d ( F i g u r e 2, T a b l e 1 ) .
D I S C U S S I O N
The present study has shown the presence of scattered at random denervated
atrophic muscle fibres and fibre type grouping, both features were found twice
in the same sample, while in the other three patients only grouped fibres of
either type I or type II were seen.
These observations strongly suggest denervation associated with reservation
2,
nevertheless these findings do not imply an associated clinical symptomatology,
for fibre type grouping can be the only finding in clinically uninvolved muscles
when there has been an adequate compensation in a chronic denervation.
It should be mentioned that in a previous study Laguens et cã.
3found
in the human biceps of EVI-positive chagasic patients non specific muscle
histo-logical changes, namely isolated hyalinazed fibres, increased number of central
nuclei in some fibres and occasionally sprouting buds containing numerous
nuclei. They did not find alterations of succinate dehydrogenase and ATPase
activities and no clear distinction between fibre types could be done by employing
the phosphorilase technique.
The changes observed in our histological study, which resulted from
dener-vation, do not point out the site of lesion within the neurogenic component
of the m.u., for either the axon or the motoneurone soma may bring about
the same histological picture. However, the electrophysiological evidences
presented by Sanz et al.
5330 ARQ- NBURO-PBJQUIATRIA (BAO PAULO) VOL. 36, Ν * k, DEZEMBRO, 1978
lesion. Their ideas agree with our findings in two aspects: first the observed
muscle changes may be the result of motoneurone soma involvement and second,
the described anatomical features can be found in* partially denervated subjects
who have recovered from a denervatory disease and, eventually, have not any
clinical symptomatology.
In conclusion, the present investigation has confirmed electrophysiological
studies in which an involvement of the m.u. was reported in patients with
chronic Chagas' disease
4'
5. Because the approach applied here has been a
morphological rather than a physiological one. it has had the advantage of
showing anatomical evidences of muscle denervation which are compatible with
an old and well compensated denervatory process which, at the time of the
investigation, may be below the clinical level of detection.
S U M M A R Y
Gastrocnemius muscle biopsies were performed in 7 subjects with chronic
Chagas' disease. On clinical and laboratory grounds the selected patients were
judged to be healthy, being the only abnormality found the presence of positive
serum tests for Chagas' disease.
Fibre type grouping of either type I or type II was observed in 5 of the 7
patients. Furthermore, in 2 of the 5 patients showing muscle fibre groupings,
angular fibres reacting with NADH and non-specific sterase were also found.
These observations strongly suggest denervation associated with reinervation.
This picture often can be observed in the skeletal muscle of patients with well
compensated denervatory conditions who do not show clinical evidences of
denervation.
R E S U M E N
Cambios histológicos e histoquimicos del músculo esquelético en la enfer¬
medad de Chagas crónica.
R E F E R E N C E S
1. C A M P B E L L , M. J . ; McCOMAS, A. J . A P E T I T O , F . —- P h y s i o l o g i c a l c h a n g e s In a g e i n g m u s c l e s . J . N e u r o l . N e u r o s u r g . P s y c h i a t . ( L o n d o n ) 36:174. 1973.
2 . K A R P A T I , G. & E N G E L , W . K . — T y p e g r o u p i n g i n s k e l e t a l m u s c l e a f t e r e x p e r i m e n t a l r e i n e r v a t i o n . N e u r o l o g y ( M i n n e a p o l i s ) 18:447, 1968.
3 . L A G U E N S , R. P . ; COSSIO, P . M . ; D I E Z , C ; S E G A L . Α . ; VAZQUEZ, C ; K R E U T ¬ Z E R , E . ; K H O U R Y , E . & A R A N A. R. Μ — I n m u n o p a t h o l o g i c a n d m o r p h o l o g i c s t u d i e s of s k e l e t a l m u s c l e i n C h a g a s ' d i s e a s e . A m J . P a t h o l o g y ( C h i c a g o ) 80:163.
1975.
4 . P A G A N O , Μ . Α . ; A R I S T I M U Ñ O . G. G.; B A S S O . S U S A N A ; COLOMBI, A. & S I C A . R. E . P . — E l e c t r o m y o g r a p h i c a l f i n d i n g s i n c h r o n i c C h a g a s ' d i s e a s e . A r a . N e u r o ¬ P s i q u i a t . ( S ã o P a u l o ) 36:316, 1978.
5 . S A N Z , OLGA, P . ; R A T U S N U . A . F . : A R I S T l M U Ñ O . G. G.; O ' N E I L L , Ε . M. & SICA, R. E . P . — A n e l e c t r o p h y s i o l o g i c a l i n v e s t i g a t i o n of s k e l e t a l m u s c l e i n h u m a n c h r o n i c C h a g a s ' d i s e a s e . A r q . N e u r o - P s i q u i a t . ( S ã o P a u l o ) 36:319, 1978.
6. SICA, R. E . P . ; SANZ, OLGA P . & COLOMBI, A. — T h e e f f e c t of a g e i n g u p o n t h e h u m a n s o l e u s m u s c l e . M e d i c i n a ( B u e n o s A i r e s ) 36:443, 1976.
R e p r i n t r e q u e s t : Dr.. R. E. P. S I C A — Sección de E l e c t r o n e u r o f i s i o l o g i a Clinica ·—
