i a m i c r o p i p e t t e technique. S a l i e n t r e s u l t s i n c l u d e d : 1 ) a 300% i n c r e a s e i n membrane v i s c o s i t y o f aged c e l l s f o l l o w i n g ionophore t r e a t m e n t corresponding t o e l e v a t e d Hb, l e v e l s ; 2 ) no change i n nembrane v i s c o s i t y and Hh, l e v e l s o f young c e l l s a f t e r ionophore exposure; 3) no dependen& between e l a s t i c shear modulus and i n t r a c e l l u l a r c a l c i u m c o n c e n t r a t i o n . 4 ) Comparison t o o s m o t i c a l l y shrunk c e l l s r e v e a l s an a d d i t i o n a l c a l c i u m c o n c e n t r a t i o n dependent a s s o c i a t i o n o f hemoglobin t o t h e membrane. This i m p l i e s nembrane mechanical p r o p e r t i e s a r e independent f r o m t h e i n t r a c e l l u l a r v i s c o s i t y . Our r e s u l t s appear r e l e v a n t t o hemolytic diseases c h a r a c t e r i z e d by e l e v a t e d (Ca) , (e.g. s i c k l e c e l l anemia) and t o age associated changes i h deformabi 1 i t y . Supported 3y DFG F r 752/1-1 u. 1-2 by NIH g r a n t s HL 15162 and HL 15722 and >y AHA-GLAA award 537IG.
ANAPHYLATOXIN ~3 a -d e s~r g . AN0 COMPLEMENT ACTIVATION PRODUCTS OF THE ALTERNATIVE AND CLASSICAL PATHWAS I N NEONATES133 G. z r~o w , M. STUTE, O. LINDERKAMP, E.P. ZILOW
U n i v e r s i t y o f Heidelberg, I n s t i t u t e o f Immunology and Department o f P e d i a t r i c s , Heidel berg, FRGThe l e v e l s of complement a c t i v a t i o n products were measured i n EOTA-pl asma o f term and preterm neonates w i t h g e s t a t i o n a l age of31-12 weeks. The a n a p h y l a t o x i n C3a-desArg was q u a n t i t a t e d w i t h a nova1 ELISA system u s i n g monoclonal antibody r e a c t i n g s e l e c t i v e l y w i t h a neoepitope o f C3a-desArg.ClrsCl1nativator and C3b(Bb)P which are generated o n l y upon a c t i v a t i o n o f t h e c l a s s i c a l o r a l t e r n a t i v e pathway, were determined by double sandwich ELISA systems. Plasma c o n c e n t r a t i o n o f C3a-desArg o f normal neonates (180 91ng/ml) d i d n o t d i f f e r s i g n i f i c a n t l y from a c o n t r o l group o f a d u l t s (171c76 ng/ml ) , w h i l e C3-levels were s i g n i f i c a n t l y decreased compared t o a d u l t s . C3a-desArg l e v e l s i n c o r d b l o o d were markedly d i m i n i s h e d (85+64ng/ml), compared t o venous blood, whereas C3 and t h e a c t i v a t i o n products C l r s C l I n a c t i v a t o r and C3(Bb)P showed no d i f f e r e n c e s between c o r d b l o o d and blood obtained by venipuncture. Neonates w i t h amniotic i n f e c t i o n as w e l l as p a t i e n t s w i t h e a r l y onset sepsis showed an i n i t i a l m a n i f o l d increase o f C3a-desArg due t o a l t e r n a t i v e pathway a c t i v a t i o n . P a t i e n t s w i t h p e r i n a t a l asphyxia had o n l y s l i g h t l y e l e v a t e d C3a-desArg l e v e l s . Thus, C3a-desArg seems t o be a h i g h l y s e n s i t i v e and s p e c i f i c i n d i c a t o r f o r severe n...
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