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Scanning electron microscopy (SEM) in haemolysis
Abstracts of The )(IV Congress or the Italian Society of Electron Micro~'copy
160
i n d i c a t e the t a l c properly assessed the e n v i r o n m e n t . 1. 2.
p o w d e r s as n o t y e t a s b e s t o s c a r r i e r s in
M. R. Becklake, An~. Rev. Respir. Dis. 114 (1976) 187-227. L. Paoletti, S. Caiazza, E. Chessa, S. Notargiacomo and G. Donelli, Ann. Ist. Sup. Sanit& 18 (1982) 341-50.
SCANNING ELECTRON IN H A E M O L Y S I S
MICROSCOPY
G.
Ricci*
Zavagli
and
G.
(SEM)
Istituto di P a t o l o g i a Speciale Medica, Universita di F e r r a r a ; *Istltuto dl C l i n i c a Medica Generale, Universita di F e r r a r a , Ferrara, Italg
Fixation procedures (Bessis a n d Weed, 1972) ] for the S E M o b s e r v a t i o n of e r y t h r o c y t e s n o w a d a y s p r o v e r e l i a b l e , so t h a t m o s t a r t i f a c t s are p r e v e n t e d from occurring. T h u s S E M r e s u l t s in a u s e f u l m e t h o d for e x a m i n i n g normal and haemolytic patterns. Normal reticulocytes are p o l y l o b u l a t e s i n c e t h e y u n d e r g o t h r o u g h the splenic slits shape remodeling a n d foreign body extrusion. In p r e m a t u r e inf a n t s the l a t t e r m a y o c c u r s p o n t a n e o u s ly t h r o u g h o u t the c i r c u l a t i o n o w i n g to spleen immaturity. Spherocytosis is d u e to A T P d e p l e t i o n in the o l d e s t c e l l s a n d thus r e p r e s e n t s the n o r m a l p r e - l y t i c stage. Such deform a t i o n is p r e s e n t in m o s t e r y t h r o c y t e s d u r i n g the h a e m o l y t i c b o u t s of h e r e d i tary spherocytosis a n d in m o s t d e f e c t s of a n a e r o b i c g l y c o l y s i s . Like hereditary spherocytosis, also elliptocytosis is c a u s e d b y p r o t e i n n ~ m b r a n e d e f e c t s . T h e h a l l m a r k of t h a l a s s a e m i a s is anisopoikilocytosis, due to i n c r e a s e d membrane-haemoglobin c o n t e n t ratio. To overlapping u n b a l a n c e of the g l o b i n chain synthesis ratio, more marked surface deformations in a l p h a - t h a n in beta-thalassaemia correspond. In Hb H disease chain excess precipitates peripherally, in C o o l e y ' s a n a e m i a c e n t r a l l y as l a r g e c l u s t e r s . T h i s is d u e to the more hydrophylic conditions of b e t a chains ( Z a v a g l i et al., 1982) 2 . Thalassaemia m a y be a s s o c i a t e d w i t h other abnormal haemoglobins, s u c h as Hb S. Haemolysis m a y a l s o be d u e to p h y s i cal agents, namely shearing stress c a u s e d b y (a) f i b r i n d e p o s i t i o n througho u t the m i c r o c i r c u l a t i o n , s u c h as in
Moschkowitz and Gasser syndromes, (b) p r o s t h e t i c h e a r t v a l v e s , a n d (c) march haemoglobinuria. IgG-caused haemolytic a n a e m i a m a y or m a v n o t be a s s o c i a t e d w i t h c o m p l e m e n t involvement: early stages may occur intravascularly, b u t l y s i s is c o m p l e t e d in the s p l e e n a n d l i v e r t h r o u g h p h a g o cytic receptors. Conversely paroxysmal noctural haemoglobinuria (PNH) is exclusively complement-mediated owing to the c h a r a c t e r i s t i c red cell suscept i b i l i t y to s u c h l y t i c s y s t e m . Scanning can reliably distinguish P N H II f r o m P N H III c e l l s ( Z a v a g l i et al., 1978) 3 . I.
M. Bessis and R. I. Weed, "Preparation of red blood cells for SEM: a survey of various artifacts" in: Scanning Electron Microscopy", O. Johari and I. Corvin, eds. IIT Research Institute, Chicago, 1972, pp. 285-94.
2.
G. Zavagli, G. Ricci, G. Castaldi, M. Marchini and P. P. Castellani, "Why inclusion bodies do assume different locations in thalassaemic erythrocytes", Blut 44 (1982) 165-72.
3.
G. Savagli, G. D. Grusovin and G. P. Guidetti, "Survival and significance of PNH erythrocytes: A scanning study", Acta Haematol. 59 (1978) 293-301.
ELECTRON MICROSCOPE D E T E C T I O N OF V I R U S LIKE PARTICLES IN B L O O D P R O D U C T S I M P L I CATED FOR NON-A, NON-B HEPATITIS E. G.
F a l c i e r i , A. A l b e r t i * , F. T r e m o l a d a * , R e a l d i * a n d C. A. B u s a c h i * * T s t i t u t o di M i c r o s c o p i a Elettronica Clinica, Universit~ di B o l o g n a ; *Istituto di M e d i c i n e Clinica, Cattedra di P a t o l o g i a Medica, UnJversita di P a d o v a ; **Istituto di P a t o l o g ~ a Medica i, Universita di B o l o g n a
The ethiological a g e n t of the n o n - A , non-B hepatitis has not yet been well characterized, but particular viruslike s t r u c t u r e s h a v e b e e n d e s c r i b e d in the n u c l e i of the h e p a t o c y t e s ]-3 a n d in the s e r u m ~,5 of a f f e c t e d p a t i e n t s . It has also been observed that blood products, l i g h t c r y o p r e c i p i t a t e s and coagu l a t i o n f a c t o r s are f r e q u e n t l y respons i b l e for n c n - A , n o n - B h e p a t i t i s in receiving subjects. In a p r e v i o u s study 6 we described non-A, non-B hepatitis in
160
i n d i c a t e the t a l c properly assessed the e n v i r o n m e n t . 1. 2.
p o w d e r s as n o t y e t a s b e s t o s c a r r i e r s in
M. R. Becklake, An~. Rev. Respir. Dis. 114 (1976) 187-227. L. Paoletti, S. Caiazza, E. Chessa, S. Notargiacomo and G. Donelli, Ann. Ist. Sup. Sanit& 18 (1982) 341-50.
SCANNING ELECTRON IN H A E M O L Y S I S
MICROSCOPY
G.
Ricci*
Zavagli
and
G.
(SEM)
Istituto di P a t o l o g i a Speciale Medica, Universita di F e r r a r a ; *Istltuto dl C l i n i c a Medica Generale, Universita di F e r r a r a , Ferrara, Italg
Fixation procedures (Bessis a n d Weed, 1972) ] for the S E M o b s e r v a t i o n of e r y t h r o c y t e s n o w a d a y s p r o v e r e l i a b l e , so t h a t m o s t a r t i f a c t s are p r e v e n t e d from occurring. T h u s S E M r e s u l t s in a u s e f u l m e t h o d for e x a m i n i n g normal and haemolytic patterns. Normal reticulocytes are p o l y l o b u l a t e s i n c e t h e y u n d e r g o t h r o u g h the splenic slits shape remodeling a n d foreign body extrusion. In p r e m a t u r e inf a n t s the l a t t e r m a y o c c u r s p o n t a n e o u s ly t h r o u g h o u t the c i r c u l a t i o n o w i n g to spleen immaturity. Spherocytosis is d u e to A T P d e p l e t i o n in the o l d e s t c e l l s a n d thus r e p r e s e n t s the n o r m a l p r e - l y t i c stage. Such deform a t i o n is p r e s e n t in m o s t e r y t h r o c y t e s d u r i n g the h a e m o l y t i c b o u t s of h e r e d i tary spherocytosis a n d in m o s t d e f e c t s of a n a e r o b i c g l y c o l y s i s . Like hereditary spherocytosis, also elliptocytosis is c a u s e d b y p r o t e i n n ~ m b r a n e d e f e c t s . T h e h a l l m a r k of t h a l a s s a e m i a s is anisopoikilocytosis, due to i n c r e a s e d membrane-haemoglobin c o n t e n t ratio. To overlapping u n b a l a n c e of the g l o b i n chain synthesis ratio, more marked surface deformations in a l p h a - t h a n in beta-thalassaemia correspond. In Hb H disease chain excess precipitates peripherally, in C o o l e y ' s a n a e m i a c e n t r a l l y as l a r g e c l u s t e r s . T h i s is d u e to the more hydrophylic conditions of b e t a chains ( Z a v a g l i et al., 1982) 2 . Thalassaemia m a y be a s s o c i a t e d w i t h other abnormal haemoglobins, s u c h as Hb S. Haemolysis m a y a l s o be d u e to p h y s i cal agents, namely shearing stress c a u s e d b y (a) f i b r i n d e p o s i t i o n througho u t the m i c r o c i r c u l a t i o n , s u c h as in
Moschkowitz and Gasser syndromes, (b) p r o s t h e t i c h e a r t v a l v e s , a n d (c) march haemoglobinuria. IgG-caused haemolytic a n a e m i a m a y or m a v n o t be a s s o c i a t e d w i t h c o m p l e m e n t involvement: early stages may occur intravascularly, b u t l y s i s is c o m p l e t e d in the s p l e e n a n d l i v e r t h r o u g h p h a g o cytic receptors. Conversely paroxysmal noctural haemoglobinuria (PNH) is exclusively complement-mediated owing to the c h a r a c t e r i s t i c red cell suscept i b i l i t y to s u c h l y t i c s y s t e m . Scanning can reliably distinguish P N H II f r o m P N H III c e l l s ( Z a v a g l i et al., 1978) 3 . I.
M. Bessis and R. I. Weed, "Preparation of red blood cells for SEM: a survey of various artifacts" in: Scanning Electron Microscopy", O. Johari and I. Corvin, eds. IIT Research Institute, Chicago, 1972, pp. 285-94.
2.
G. Zavagli, G. Ricci, G. Castaldi, M. Marchini and P. P. Castellani, "Why inclusion bodies do assume different locations in thalassaemic erythrocytes", Blut 44 (1982) 165-72.
3.
G. Savagli, G. D. Grusovin and G. P. Guidetti, "Survival and significance of PNH erythrocytes: A scanning study", Acta Haematol. 59 (1978) 293-301.
ELECTRON MICROSCOPE D E T E C T I O N OF V I R U S LIKE PARTICLES IN B L O O D P R O D U C T S I M P L I CATED FOR NON-A, NON-B HEPATITIS E. G.
F a l c i e r i , A. A l b e r t i * , F. T r e m o l a d a * , R e a l d i * a n d C. A. B u s a c h i * * T s t i t u t o di M i c r o s c o p i a Elettronica Clinica, Universit~ di B o l o g n a ; *Istituto di M e d i c i n e Clinica, Cattedra di P a t o l o g i a Medica, UnJversita di P a d o v a ; **Istituto di P a t o l o g ~ a Medica i, Universita di B o l o g n a
The ethiological a g e n t of the n o n - A , non-B hepatitis has not yet been well characterized, but particular viruslike s t r u c t u r e s h a v e b e e n d e s c r i b e d in the n u c l e i of the h e p a t o c y t e s ]-3 a n d in the s e r u m ~,5 of a f f e c t e d p a t i e n t s . It has also been observed that blood products, l i g h t c r y o p r e c i p i t a t e s and coagu l a t i o n f a c t o r s are f r e q u e n t l y respons i b l e for n c n - A , n o n - B h e p a t i t i s in receiving subjects. In a p r e v i o u s study 6 we described non-A, non-B hepatitis in