56
Specialia
EXPERIENI"IA31/1
Table iII. Effect of cupric ions on experimental oral therapy with cephaloglycin and SK & F 44065 in mouse infected with E. coli Compound
Cephaloglycin
S K & F 44065
Dissolved in (and administered oraIly)
Saline CuSO 4 solution (1 [zg/ml) D-Penieillamine (euprimine) (5 ~xg/ml) CuSO 4 (1 txg/ml) + cuprimine (5 [zg/ml) Saline CuSO 4 solution (1 p.g/ml) D-Penicillamine (cuprimine) (5 [xg/ml) CuSQ (1 b~g/mI) + cuprimine (5 ~g/ml)
T h e d a t a in T a b l e 11 d e m o n s t r a t e t h a t c e p h a l o g l y c i n a n d S K & F 44065 [ 3 - ( 5 - m e t h y l - l , 3 , 4 - t h i a d i a z o l - 2 y l t h i o m e t h y l ) - 7 - ( p h e n y l g l y c i n e a c e t a m i d o ) - 3-cephem-4c a r b o x y l i c acid], a n o t h e r p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n b e h a v e similarly. T h e d e g r a d i n g effect of i n c r e a s i n g c o n c e n t r a t i o n s of cupric ion a n d t h e n e u t r a l i z a t i o n of t h i s effect b y d i f f e r e n t c h e l a t i n g a g e n t s is s h o w n for b o t h cephalosporins. T h e r a t e of h y d r o l y z i n g effect of cupric ion o n v a r i o u s p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n s was f o u n d to be a f u n c t i o n of t h e s u b s t i t u e n t a t t h e 3-position of t h e c e p h a l o s p o r i n n u c l e u s a n d also was i n f l u e n c e d b y substitution on the phellylglycine moiety. The results o b t a i n e d w i t h a larger n u m b e r of p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n s will be p u b l i s h e d in d e t a i l elsewhere. T h e d e g r a d i n g effect of CuSO4 o n t h e a n t i n l i c r o b i a l a c t i v i t y of c e p h a l o g l y c i n a n d S K & F 44065 was f o u n d also in m o u s e in a n i n f e c t i o n - p r o t e c t i o n a s s a y w i t h E. colt (Table 111) T h e EDs0 v a l u e s of t h e 2 c o m p o u n d s are s i g n i f i c a n t l y i n c r e a s e d (loss of p o t e n c y ) w h e n t h e y were dissolved in CuSO 4 s o l u t i o n (1 ~g/ml) a n d adm i n i s t e r e d orally to mice. T h e s i m u l t a n e o u s a d d i t i o n of D-penicillamine is able t o c o u n t e r a c t t h i s effect of cupric ion.
EDs0 (mg/kg) Exp. 1 6.2 48 22 12 7.5 35 9 9.2
Exp. 2 15.5 42 --9.5 25 ---
T h e s e e x p e r i m e n t s clearly d e m o n s t r a t e t h a t t h e orally active phenylglycine-type semisynthetic cephalosporins ( b u t n o t t h e o t h e r types) are i n a c t i v a t e d b y cupric ion as are t h e penicillins. T h e i m p l i c a t i o n s of t h e s e f i n d i n g s t o u c h several a s p e c t s of c e p h a l o s p o r i n r e s e a r c h i n c l u d i n g bioassay, s t a b i l i t y , f o r m u l a t i o n a n d e x p e r i m e n t a l a n d clinical trials.
Rdsumd. T o u t e s les p6nicillines s o n t d6grad6es p a r les ions de cuivre a v e c la f o r m a t i o n d ' a c i d e p6nicilloique. P a r r o t les c 6 p h a l o s p o r i n e s d e m i s y n t h 6 t i q u e s seuls les d6riv6s c o n t e n a n t d u p h 6 n y l g l y c i n e s o n t sensitives l'effet d 6 g r a d a n t d u cuivre. L a D , p 6 n i c i l l a m i n e est c a p a b l e de le c o n t r e c a r r e r . j . v . URI, P. ACTOR, L. PHILLIPS
a n d J. A. WEISBACH
Smith Kline 6, French Laboratories, 7500 Spring Garden Street, Philadelphia (Pennsylvania 19101, U S A ) , 29 J u l y 1974.
Frequency of Hypocatalasemia in a Sample of Spanish Population A c c o r d i n g t o NISHIMURA et aI. 1, i n d i v i d u a l s w h o s e c a t a l a t i c a c t i v i t y in b l o o d is b e l o w t h e n o r m a l r a n g e m u s t b e c o n s i d e r e d as h y p o c a t a l a t i c s . AEBI et a l * h a v e . d e t e c t e d in a screening s t u d y a c t i v i t i e s v a r y i n g f r o m 16 t o 85% of t h e n o r m a l levels. I n v e s t i g a t i o n s c a r r i e d o u t b y TAKAHARA et al.8; HAMILTON et al.4; AEBI et al. ~ a n d BAUR 6, suggest t h a t one or m o r e a u t o s o m i c c o d o m i n a n t genes w i t h v a r i a b l e e x p r e s s i v i t y i n c l u d i n g 0 (recessive) are r e s p o n s i b l e for t h i s defect (TAI~AI-IARAet al. a). So far, d a t a o n t h e f r e q u e n c y of t h i s deficiency in d i f f e r e n t h u m a n g r o u p s are v e r y scarce. T h e r e are a v a i l a b l e d a t a o n l y o n p o p u l a t i o n s f r o m Asia (TAKAHARA 7 in AEBI s) S w i t z e r l a n d (AEBI 2) a n d a J e w i s h f a m i l y f r o m I r a n (SzEINBERC et al~ s). T h e o b j e c t of t h i s s t u d y is t h e d e t e r m i n a t i o n of h y p o c a t a l a s e m i a f r e q u e n c y in a s a m p l e of t h e s p a n i s h p o p u l a tion. A t o t a l of 1%009 s p e c i m e n s h a v e b e e n s a m p l e d f r o m subjects visiting the Ambulatorio 'Pedro Gonzalez B u e n o ' , all of t h e m l i v i n g ill t h e E a s t e r n a r e a s of Madrid, m o s t of t h e m i m m i g r a n t s f r o m o t h e r s p a n i s h p r o v i n c e s (77% f r o m S o u t h e r n a n d 1 7 % f r o m t h e N o r t h e r n
Castilian plateau and 6% from the surrounding provinces or e v e n of m i x e d origins). Methods. A s a m p l e of 2 m l of b l o o d fronl e v e r y s u b j e c t was m i x e d w i t h 0.5 m l of a 3.8% c i t r a t e solution. Before a n a l y s i s t h e b l o o d was k e e p t a t r o o m t e m p e r a t u r e for
1 2. T. NISHIMURA, H. B. t~AMILTON, T. Y. KOBARA, S. TAKAIIARA, Y. OOURA and K. DoI, Science 730, 333 (1959). g H. AEBI, V. JEUNET, R. RICHTERICH, H. SUTER, R. B1JTLER,
J. FREI and H. R. MARTI, Enzym. Biol. Clin. 2, 1 (1962/63). 3 S. TAKAHARA, H. B. HAMILTON, J. V. NEEL, T. Y.' KOBARA, Y. OGURA, E. Z. HISHIMURA, g . 0ZAKI and K. ITO, J. din. Invest. 39, 610 (1960). r H. B. HAMILTON, J. V. NEEL, T. Y. KOBARA and K. OZAKI
J. elin. Invest. dO, 2199 (1961). H. AEBI, in Advances in Human Genetics (Eds H. HARRIS and HIRSCHORN; Plenum Press. New York/London 1971), vol. 2, p. 143 6 E. W. BAUR, Science 150, 816 (1963).
7 S. TAKAHARAand H. MIYAMOTO,Otorhinolaring. Clin., Jibiinoka 21, 2 (1949). 8 A. SZEINBERG, A. DE VRIES, J. PINKHAS, M. DJALDETTI and R. EZRA, Acta Genet. Ined. Gemell. 72, 247 (1963).
15. 1. 1975
Speeialia
n o t m o r e t h a n 6 h. F o r d e t e r m i n a t i o n of c a t a l a t i c a c t i v i t y FEINSTEIN'S ~ p e r b o r a t e m e t h o d w i t h s l i g h t m o d i f i c a t i o n s was used. T h e s c r e e n i n g p r o c e d u r e was carried o u t b y a d d i n g 0.003 m l b l o o d t o 5 m l 2.5% p e r b o r a t e solution. T h o s e cases, u n a b l e t o d e c o m p o s e t h i s a m o u n t of p e r b o r a t e w i t h i n 10 m i n , were f u r t h e r e v a l u a t e d b y u s i n g d e c r e a s i n g a m o u n t s (4, 3, 2, a n d 1 ml) of t h e s a m e solution. T h i s p r o c e d u r e p e r m i t s t h e d e t e c t i o n of a c t i v i t y levels corresp o n d i n g t o i n t e r v a l s of 0-20 ; 20-40 ; 4 0 - 6 0 ; a n d 6 0 - 8 0 % ol t h e n o r m a l level. N o b e t t e r a c c u r a c y w a s a t t e m p t e d for t h e p u r p o s e of t h i s s t u d y . A g r o u p of 20 n o r m a l s u b j e c t s was s t u d i e d b y t h i s m e t h o d . T h e i r a v e r a g e c a t a l a s e a c t i v i t y w a s 0.526 =k 0.018 ( X • 5 ) e x p r e s s e d as m21~rof decomposed perborate. Results. A m o n g t h e 10,009 b l o o d s a m p l e s of d i f f e r e n t i n d i v i d u a l s , 26 were f o u n d t o h a v e a n e n z y m a t i c a c t i v i t y b e l o w t h e n o r m a l range. T h e y were d i s t r i b u t e d as follows : 2 0 - 4 0 % i n t e r v a l 4 s a m p l e s ; 4 0 - 6 0 % i n t e r v a l 20 s a m p l e s a n d in t h e 6 0 - 8 0 i n t e r v a l 2 samples. T h e p e r c e n t a g e of h y p o c a t a l a s e m i a is t h e r e f o r e 0.26%, w h i c h m e a n s one p e r 384 i n d i v i d u a l s . If we t a k e all tile cases t o g e t h e r , n o t c o n s i d e r i n g t h e p o s i b i l i t y of t h e e x i s t e n c e of several alleles, t h e f r e q u e n c y w o u l d be 0.0013, w i t h a c o n f i d e n c e l i m i t of 0.008 t o 0.0019. W e h a v e n o t f o u n d cases of a c a t a l a s e m i a ( t e r m coined b y TAKAHARA a n d MIYAMOTOT), a n d o n l y one of t h e i n d i v i d u a l s was affected w i t h p y o r r h e a , a w o m a n w h o s e c a t a l a t i c a c t i v i t y t u r n e d o u t to be lowest ill t h i s screening. A d a u g h t e r of t h i s w o m a n was also affected w i t h t h e s a m e illness, b u t h e r c a t a l a t i c a c t i v i t y was n o r m a l . Discussion. T h e d a t a a v a i l a b l e do n o t allow one t o decide to w h i c h t y p e of AEBI'S ~ classification t h e hypo_ c a t a l a s e m i a cases f o u n d in S p a i n m a y b e a t t r i b u t e d . T h e v a r i a b i l i t y of c a t a l a s e a c t i v i t y f o u n d in t h i s p o p u l a t i o n m a y b e of g e n e t i c origin, such as t h e p r e s e n c e of several alleles, or i t m a y b e due t o e x p e r i m e n t a l v a r i a t i o n since t h e n u m b e r of cells e m p l o y e d for t h i s a n a l y s i s was n o t e x a c t l y d e t e r m i n e d . I n cases of ferropenic a n d h a e m o l y t i c a n a e m i a , n o r m a l v a l u e s are f o u n d w h e n c a t a l a t i c a c t i v i t y is r e l a t e d to h a e m o g l o b i n c o n t e n t (PAUL a n d ENCSrED ~0).
57
I t is for t h i s r e a s o n t h a t , w h e n m e a s u r i n g a b s o l u t e values, t h e s e s u b j e c t s m a y a p p e a r as h y p o c a t a l a t i c . I t w a s assumed that the haemoglobin concentration would be a p p r o x i m a t e l y n o r m a l in all t h e s a m p l e s a n a l y z e d . I t m a y also b e t h a t h y p o c a t a l a t i c s u b j e c t s h a v e b e e n overlooked. If t h e r e is all o v e r l a p b e t w e e n t h e a c t i v i t i e s of hypocatalatics and normals, those individuals with an a c t i v i t y of o v e r 8 0 % m a y h a v e b e e n scored as n o r m a l s . D u e t o t h i s o v e r l a p p i n g , it is possible t h a t o u r figures u n d e r e s t i m a t e t h e real n u m b e r of h y p o c a t a l a t i c s (i.e. in cases of g r o u p I I I b deficiency, AEBIs). O n t h e o t h e r h a n d , t h e p r e s e n c e of e n z y m e i n h i b i t o r s in b l o o d s e r u m , as d e m o n s t r a t e d b y PAUL a n d ENGSTED 10, m a y c o n t r i b u t e to an overstimation. T h e figure for t h e f r e q u e n c y of t h e a c a t a l a s e m i a gene (0.0013) p r o b a b l y r e p r e s e n t s a m a x i m a l value. N e v e r theless o u r v a l u e s are in a g r e e m e n t w i t h t h e a s s u m p t i o n t h a t - a l t h o u g h t h e a v e r a g e gene f r e q u e n c y seems t o v a r y f r o m c o u n t r y to c o u n t r y (TAKAHARA 1967 in AEBIS), t h e a c a t a l a s e m i a gene is of w o r l d w i d e d i s t r i b u t i o n .
Resumen. Se h a e s t u d i a d o la a c t i v i d a d catalAsica e n la s a n g r e de 10.009 espafioles r e s i d e n t e s en Madrid, p o t el m d t o d o de FEIIqSTEINt Se e a c o n t r a r o n 26 i u d i v i d u o s c u y a a c t i v i d a d catalAsica oscila e n t r e el 2 0 % y el 80~ de la n o r m a l . E s t o s i n d i v i d u o s se c o n s i d e r a r o n como h i p o c a t a l a s 6 m i c o s . Se c o m p a r a n estos d a t o s con los de otros a u t o r e s y se d i s c u t e su posible significado. A. PELLICER, L. VICENTE, }~NCARNACION LOPEZ, MERCEDES D~ LAS HERAS a n d ANGIELA CASADO
[nstitulo de Gendlica y A~tropologia C.S.I.C., Velazquez 144, Madrid-6 (Spain), 22 July 1974.
9 R. N. FEINSTEIN,J. B. HOWARD,L. B. BALLONOFF and J. E. SEAHOLM,Annalyt. Biochem. 8, 277 (1964). a0 K. G. PAUL and L. M. E~GSTED, Scand. J. clin. Lab. Invest. 10, 26 (1958).
Difference in Mechanical Properties of Adjacent Sarcomeres in Single Striated Muscle Fibres of the Horseshoe Crab (Tachypleus gigas) S t r i a t e d m u s c l e fibres c o n t a i n b o t h c o n t r a c t i l e a n d visco-elastic e l e m e n t s . S u c h visco-elastic e l e m e n t s could b e m a d e t o r e v e a l t h e m s e l v e s w h e n t h e m u s c l e is overl o a d e d i.e. u n d e r so-called i s o m e t r i c c o n d i t i o n s . HILL * suggested that during isometric contraction the actively c o n t r a c t i l e e l e m e n t s w o u l d s h o r t e n a t t h e e x p e n s e of t h e p a s s i v e elastic e l e m e n t s . Since t h e n , s e v e r a l m e t h o d s h a v e b e e n d e s i g n e d t o t e s t his s u g g e s t i o n ~-6. H o w e v e r all t h e m e t h o d s used t o m e a s u r e t h e p r o p e r t i e s of t h e elastic e l e m e n t s were b a s e d on e s t i m a t i o n s f r o m t h e forcevelocity relation and the relation between tension and t i m e d e r i v a t i v e of i s o m e t r i c t e n s i o n d u r i n g t e t a n u s L I n t h i s p a p e r we h a v e used t h e m e t h o d of s i m u l t a n e o u s m e a s u r e m e n t s of a d j a c e n t s a r c o m e r e l e n g t h c h a n g e s t o s t u d y t h e b e h a v i o u r s of a d j a c e n t s a r c o m e r e s in single s t r i a t e d m u s c l e f i b r e s d u r i n g i s o m e t r i c t w i t c h s~,. T h e o b s e r v e d difference in t h e a v e r a g e velocities of a d j a c e n t Z lines in some special cases is c o n s i d e r e d to b e a n i n d e x of t h e difference in t h e visco-elastic e l e m e n t s of a d j a c e n t sarcomeres.
Materials and methods. T h e e x p e r i m e n t s were p e r f o r m e d o n t h e s h o r t accessory muscle (SAM), a r e c e p t o r m u s c l e sb, in t h e w a l k i n g leg of t h e A s i a t i c h o r s e s h o e crab, Tachypleus gigas. T h e s e muscle fibres were selected for s t u d y b e c a u s e of t h e i r b r o a d s t r i a t i o n s (2-12 ~m) a n d s m a l l d i a m e t e r s
1 A. V. HILL, Proc. R. Soc. B726, 136 (1938). 2 B. I{ATZ,J. Physiol., Lond. 95, 45 (1939). a A. V. HILL, Proc. R. Soc. B 136, 399 (1949). 4 D. R. WILKIE, J. Physiol., Lond. 110, 249 (1950). ]3. R. JEWELL and D. R. WILKIE, J. Physiol., Loud. 143, 515 (1958). H. L. MCCROREY, H. H. GALE and N. R. ALPERT,Am. J. Physiol. 210, 114 (1966). R. I. CLOSE, Physiol. Rev. 52, 129 (1972).
s~ y. M. CHEU~O and J. C. HWAN% Proe. Aust. physiol, pharmac. Soc. 3, 213 {1972). sb S. B. BARBER and W. F. HAYES, Comp. Bioehem. Physiol. 11, 193 (1964).
Specialia
EXPERIENI"IA31/1
Table iII. Effect of cupric ions on experimental oral therapy with cephaloglycin and SK & F 44065 in mouse infected with E. coli Compound
Cephaloglycin
S K & F 44065
Dissolved in (and administered oraIly)
Saline CuSO 4 solution (1 [zg/ml) D-Penieillamine (euprimine) (5 ~xg/ml) CuSO 4 (1 txg/ml) + cuprimine (5 [zg/ml) Saline CuSO 4 solution (1 p.g/ml) D-Penicillamine (cuprimine) (5 [xg/ml) CuSQ (1 b~g/mI) + cuprimine (5 ~g/ml)
T h e d a t a in T a b l e 11 d e m o n s t r a t e t h a t c e p h a l o g l y c i n a n d S K & F 44065 [ 3 - ( 5 - m e t h y l - l , 3 , 4 - t h i a d i a z o l - 2 y l t h i o m e t h y l ) - 7 - ( p h e n y l g l y c i n e a c e t a m i d o ) - 3-cephem-4c a r b o x y l i c acid], a n o t h e r p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n b e h a v e similarly. T h e d e g r a d i n g effect of i n c r e a s i n g c o n c e n t r a t i o n s of cupric ion a n d t h e n e u t r a l i z a t i o n of t h i s effect b y d i f f e r e n t c h e l a t i n g a g e n t s is s h o w n for b o t h cephalosporins. T h e r a t e of h y d r o l y z i n g effect of cupric ion o n v a r i o u s p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n s was f o u n d to be a f u n c t i o n of t h e s u b s t i t u e n t a t t h e 3-position of t h e c e p h a l o s p o r i n n u c l e u s a n d also was i n f l u e n c e d b y substitution on the phellylglycine moiety. The results o b t a i n e d w i t h a larger n u m b e r of p h e n y l g l y c i n e - t y p e c e p h a l o s p o r i n s will be p u b l i s h e d in d e t a i l elsewhere. T h e d e g r a d i n g effect of CuSO4 o n t h e a n t i n l i c r o b i a l a c t i v i t y of c e p h a l o g l y c i n a n d S K & F 44065 was f o u n d also in m o u s e in a n i n f e c t i o n - p r o t e c t i o n a s s a y w i t h E. colt (Table 111) T h e EDs0 v a l u e s of t h e 2 c o m p o u n d s are s i g n i f i c a n t l y i n c r e a s e d (loss of p o t e n c y ) w h e n t h e y were dissolved in CuSO 4 s o l u t i o n (1 ~g/ml) a n d adm i n i s t e r e d orally to mice. T h e s i m u l t a n e o u s a d d i t i o n of D-penicillamine is able t o c o u n t e r a c t t h i s effect of cupric ion.
EDs0 (mg/kg) Exp. 1 6.2 48 22 12 7.5 35 9 9.2
Exp. 2 15.5 42 --9.5 25 ---
T h e s e e x p e r i m e n t s clearly d e m o n s t r a t e t h a t t h e orally active phenylglycine-type semisynthetic cephalosporins ( b u t n o t t h e o t h e r types) are i n a c t i v a t e d b y cupric ion as are t h e penicillins. T h e i m p l i c a t i o n s of t h e s e f i n d i n g s t o u c h several a s p e c t s of c e p h a l o s p o r i n r e s e a r c h i n c l u d i n g bioassay, s t a b i l i t y , f o r m u l a t i o n a n d e x p e r i m e n t a l a n d clinical trials.
Rdsumd. T o u t e s les p6nicillines s o n t d6grad6es p a r les ions de cuivre a v e c la f o r m a t i o n d ' a c i d e p6nicilloique. P a r r o t les c 6 p h a l o s p o r i n e s d e m i s y n t h 6 t i q u e s seuls les d6riv6s c o n t e n a n t d u p h 6 n y l g l y c i n e s o n t sensitives l'effet d 6 g r a d a n t d u cuivre. L a D , p 6 n i c i l l a m i n e est c a p a b l e de le c o n t r e c a r r e r . j . v . URI, P. ACTOR, L. PHILLIPS
a n d J. A. WEISBACH
Smith Kline 6, French Laboratories, 7500 Spring Garden Street, Philadelphia (Pennsylvania 19101, U S A ) , 29 J u l y 1974.
Frequency of Hypocatalasemia in a Sample of Spanish Population A c c o r d i n g t o NISHIMURA et aI. 1, i n d i v i d u a l s w h o s e c a t a l a t i c a c t i v i t y in b l o o d is b e l o w t h e n o r m a l r a n g e m u s t b e c o n s i d e r e d as h y p o c a t a l a t i c s . AEBI et a l * h a v e . d e t e c t e d in a screening s t u d y a c t i v i t i e s v a r y i n g f r o m 16 t o 85% of t h e n o r m a l levels. I n v e s t i g a t i o n s c a r r i e d o u t b y TAKAHARA et al.8; HAMILTON et al.4; AEBI et al. ~ a n d BAUR 6, suggest t h a t one or m o r e a u t o s o m i c c o d o m i n a n t genes w i t h v a r i a b l e e x p r e s s i v i t y i n c l u d i n g 0 (recessive) are r e s p o n s i b l e for t h i s defect (TAI~AI-IARAet al. a). So far, d a t a o n t h e f r e q u e n c y of t h i s deficiency in d i f f e r e n t h u m a n g r o u p s are v e r y scarce. T h e r e are a v a i l a b l e d a t a o n l y o n p o p u l a t i o n s f r o m Asia (TAKAHARA 7 in AEBI s) S w i t z e r l a n d (AEBI 2) a n d a J e w i s h f a m i l y f r o m I r a n (SzEINBERC et al~ s). T h e o b j e c t of t h i s s t u d y is t h e d e t e r m i n a t i o n of h y p o c a t a l a s e m i a f r e q u e n c y in a s a m p l e of t h e s p a n i s h p o p u l a tion. A t o t a l of 1%009 s p e c i m e n s h a v e b e e n s a m p l e d f r o m subjects visiting the Ambulatorio 'Pedro Gonzalez B u e n o ' , all of t h e m l i v i n g ill t h e E a s t e r n a r e a s of Madrid, m o s t of t h e m i m m i g r a n t s f r o m o t h e r s p a n i s h p r o v i n c e s (77% f r o m S o u t h e r n a n d 1 7 % f r o m t h e N o r t h e r n
Castilian plateau and 6% from the surrounding provinces or e v e n of m i x e d origins). Methods. A s a m p l e of 2 m l of b l o o d fronl e v e r y s u b j e c t was m i x e d w i t h 0.5 m l of a 3.8% c i t r a t e solution. Before a n a l y s i s t h e b l o o d was k e e p t a t r o o m t e m p e r a t u r e for
1 2. T. NISHIMURA, H. B. t~AMILTON, T. Y. KOBARA, S. TAKAIIARA, Y. OOURA and K. DoI, Science 730, 333 (1959). g H. AEBI, V. JEUNET, R. RICHTERICH, H. SUTER, R. B1JTLER,
J. FREI and H. R. MARTI, Enzym. Biol. Clin. 2, 1 (1962/63). 3 S. TAKAHARA, H. B. HAMILTON, J. V. NEEL, T. Y.' KOBARA, Y. OGURA, E. Z. HISHIMURA, g . 0ZAKI and K. ITO, J. din. Invest. 39, 610 (1960). r H. B. HAMILTON, J. V. NEEL, T. Y. KOBARA and K. OZAKI
J. elin. Invest. dO, 2199 (1961). H. AEBI, in Advances in Human Genetics (Eds H. HARRIS and HIRSCHORN; Plenum Press. New York/London 1971), vol. 2, p. 143 6 E. W. BAUR, Science 150, 816 (1963).
7 S. TAKAHARAand H. MIYAMOTO,Otorhinolaring. Clin., Jibiinoka 21, 2 (1949). 8 A. SZEINBERG, A. DE VRIES, J. PINKHAS, M. DJALDETTI and R. EZRA, Acta Genet. Ined. Gemell. 72, 247 (1963).
15. 1. 1975
Speeialia
n o t m o r e t h a n 6 h. F o r d e t e r m i n a t i o n of c a t a l a t i c a c t i v i t y FEINSTEIN'S ~ p e r b o r a t e m e t h o d w i t h s l i g h t m o d i f i c a t i o n s was used. T h e s c r e e n i n g p r o c e d u r e was carried o u t b y a d d i n g 0.003 m l b l o o d t o 5 m l 2.5% p e r b o r a t e solution. T h o s e cases, u n a b l e t o d e c o m p o s e t h i s a m o u n t of p e r b o r a t e w i t h i n 10 m i n , were f u r t h e r e v a l u a t e d b y u s i n g d e c r e a s i n g a m o u n t s (4, 3, 2, a n d 1 ml) of t h e s a m e solution. T h i s p r o c e d u r e p e r m i t s t h e d e t e c t i o n of a c t i v i t y levels corresp o n d i n g t o i n t e r v a l s of 0-20 ; 20-40 ; 4 0 - 6 0 ; a n d 6 0 - 8 0 % ol t h e n o r m a l level. N o b e t t e r a c c u r a c y w a s a t t e m p t e d for t h e p u r p o s e of t h i s s t u d y . A g r o u p of 20 n o r m a l s u b j e c t s was s t u d i e d b y t h i s m e t h o d . T h e i r a v e r a g e c a t a l a s e a c t i v i t y w a s 0.526 =k 0.018 ( X • 5 ) e x p r e s s e d as m21~rof decomposed perborate. Results. A m o n g t h e 10,009 b l o o d s a m p l e s of d i f f e r e n t i n d i v i d u a l s , 26 were f o u n d t o h a v e a n e n z y m a t i c a c t i v i t y b e l o w t h e n o r m a l range. T h e y were d i s t r i b u t e d as follows : 2 0 - 4 0 % i n t e r v a l 4 s a m p l e s ; 4 0 - 6 0 % i n t e r v a l 20 s a m p l e s a n d in t h e 6 0 - 8 0 i n t e r v a l 2 samples. T h e p e r c e n t a g e of h y p o c a t a l a s e m i a is t h e r e f o r e 0.26%, w h i c h m e a n s one p e r 384 i n d i v i d u a l s . If we t a k e all tile cases t o g e t h e r , n o t c o n s i d e r i n g t h e p o s i b i l i t y of t h e e x i s t e n c e of several alleles, t h e f r e q u e n c y w o u l d be 0.0013, w i t h a c o n f i d e n c e l i m i t of 0.008 t o 0.0019. W e h a v e n o t f o u n d cases of a c a t a l a s e m i a ( t e r m coined b y TAKAHARA a n d MIYAMOTOT), a n d o n l y one of t h e i n d i v i d u a l s was affected w i t h p y o r r h e a , a w o m a n w h o s e c a t a l a t i c a c t i v i t y t u r n e d o u t to be lowest ill t h i s screening. A d a u g h t e r of t h i s w o m a n was also affected w i t h t h e s a m e illness, b u t h e r c a t a l a t i c a c t i v i t y was n o r m a l . Discussion. T h e d a t a a v a i l a b l e do n o t allow one t o decide to w h i c h t y p e of AEBI'S ~ classification t h e hypo_ c a t a l a s e m i a cases f o u n d in S p a i n m a y b e a t t r i b u t e d . T h e v a r i a b i l i t y of c a t a l a s e a c t i v i t y f o u n d in t h i s p o p u l a t i o n m a y b e of g e n e t i c origin, such as t h e p r e s e n c e of several alleles, or i t m a y b e due t o e x p e r i m e n t a l v a r i a t i o n since t h e n u m b e r of cells e m p l o y e d for t h i s a n a l y s i s was n o t e x a c t l y d e t e r m i n e d . I n cases of ferropenic a n d h a e m o l y t i c a n a e m i a , n o r m a l v a l u e s are f o u n d w h e n c a t a l a t i c a c t i v i t y is r e l a t e d to h a e m o g l o b i n c o n t e n t (PAUL a n d ENCSrED ~0).
57
I t is for t h i s r e a s o n t h a t , w h e n m e a s u r i n g a b s o l u t e values, t h e s e s u b j e c t s m a y a p p e a r as h y p o c a t a l a t i c . I t w a s assumed that the haemoglobin concentration would be a p p r o x i m a t e l y n o r m a l in all t h e s a m p l e s a n a l y z e d . I t m a y also b e t h a t h y p o c a t a l a t i c s u b j e c t s h a v e b e e n overlooked. If t h e r e is all o v e r l a p b e t w e e n t h e a c t i v i t i e s of hypocatalatics and normals, those individuals with an a c t i v i t y of o v e r 8 0 % m a y h a v e b e e n scored as n o r m a l s . D u e t o t h i s o v e r l a p p i n g , it is possible t h a t o u r figures u n d e r e s t i m a t e t h e real n u m b e r of h y p o c a t a l a t i c s (i.e. in cases of g r o u p I I I b deficiency, AEBIs). O n t h e o t h e r h a n d , t h e p r e s e n c e of e n z y m e i n h i b i t o r s in b l o o d s e r u m , as d e m o n s t r a t e d b y PAUL a n d ENGSTED 10, m a y c o n t r i b u t e to an overstimation. T h e figure for t h e f r e q u e n c y of t h e a c a t a l a s e m i a gene (0.0013) p r o b a b l y r e p r e s e n t s a m a x i m a l value. N e v e r theless o u r v a l u e s are in a g r e e m e n t w i t h t h e a s s u m p t i o n t h a t - a l t h o u g h t h e a v e r a g e gene f r e q u e n c y seems t o v a r y f r o m c o u n t r y to c o u n t r y (TAKAHARA 1967 in AEBIS), t h e a c a t a l a s e m i a gene is of w o r l d w i d e d i s t r i b u t i o n .
Resumen. Se h a e s t u d i a d o la a c t i v i d a d catalAsica e n la s a n g r e de 10.009 espafioles r e s i d e n t e s en Madrid, p o t el m d t o d o de FEIIqSTEINt Se e a c o n t r a r o n 26 i u d i v i d u o s c u y a a c t i v i d a d catalAsica oscila e n t r e el 2 0 % y el 80~ de la n o r m a l . E s t o s i n d i v i d u o s se c o n s i d e r a r o n como h i p o c a t a l a s 6 m i c o s . Se c o m p a r a n estos d a t o s con los de otros a u t o r e s y se d i s c u t e su posible significado. A. PELLICER, L. VICENTE, }~NCARNACION LOPEZ, MERCEDES D~ LAS HERAS a n d ANGIELA CASADO
[nstitulo de Gendlica y A~tropologia C.S.I.C., Velazquez 144, Madrid-6 (Spain), 22 July 1974.
9 R. N. FEINSTEIN,J. B. HOWARD,L. B. BALLONOFF and J. E. SEAHOLM,Annalyt. Biochem. 8, 277 (1964). a0 K. G. PAUL and L. M. E~GSTED, Scand. J. clin. Lab. Invest. 10, 26 (1958).
Difference in Mechanical Properties of Adjacent Sarcomeres in Single Striated Muscle Fibres of the Horseshoe Crab (Tachypleus gigas) S t r i a t e d m u s c l e fibres c o n t a i n b o t h c o n t r a c t i l e a n d visco-elastic e l e m e n t s . S u c h visco-elastic e l e m e n t s could b e m a d e t o r e v e a l t h e m s e l v e s w h e n t h e m u s c l e is overl o a d e d i.e. u n d e r so-called i s o m e t r i c c o n d i t i o n s . HILL * suggested that during isometric contraction the actively c o n t r a c t i l e e l e m e n t s w o u l d s h o r t e n a t t h e e x p e n s e of t h e p a s s i v e elastic e l e m e n t s . Since t h e n , s e v e r a l m e t h o d s h a v e b e e n d e s i g n e d t o t e s t his s u g g e s t i o n ~-6. H o w e v e r all t h e m e t h o d s used t o m e a s u r e t h e p r o p e r t i e s of t h e elastic e l e m e n t s were b a s e d on e s t i m a t i o n s f r o m t h e forcevelocity relation and the relation between tension and t i m e d e r i v a t i v e of i s o m e t r i c t e n s i o n d u r i n g t e t a n u s L I n t h i s p a p e r we h a v e used t h e m e t h o d of s i m u l t a n e o u s m e a s u r e m e n t s of a d j a c e n t s a r c o m e r e l e n g t h c h a n g e s t o s t u d y t h e b e h a v i o u r s of a d j a c e n t s a r c o m e r e s in single s t r i a t e d m u s c l e f i b r e s d u r i n g i s o m e t r i c t w i t c h s~,. T h e o b s e r v e d difference in t h e a v e r a g e velocities of a d j a c e n t Z lines in some special cases is c o n s i d e r e d to b e a n i n d e x of t h e difference in t h e visco-elastic e l e m e n t s of a d j a c e n t sarcomeres.
Materials and methods. T h e e x p e r i m e n t s were p e r f o r m e d o n t h e s h o r t accessory muscle (SAM), a r e c e p t o r m u s c l e sb, in t h e w a l k i n g leg of t h e A s i a t i c h o r s e s h o e crab, Tachypleus gigas. T h e s e muscle fibres were selected for s t u d y b e c a u s e of t h e i r b r o a d s t r i a t i o n s (2-12 ~m) a n d s m a l l d i a m e t e r s
1 A. V. HILL, Proc. R. Soc. B726, 136 (1938). 2 B. I{ATZ,J. Physiol., Lond. 95, 45 (1939). a A. V. HILL, Proc. R. Soc. B 136, 399 (1949). 4 D. R. WILKIE, J. Physiol., Lond. 110, 249 (1950). ]3. R. JEWELL and D. R. WILKIE, J. Physiol., Loud. 143, 515 (1958). H. L. MCCROREY, H. H. GALE and N. R. ALPERT,Am. J. Physiol. 210, 114 (1966). R. I. CLOSE, Physiol. Rev. 52, 129 (1972).
s~ y. M. CHEU~O and J. C. HWAN% Proe. Aust. physiol, pharmac. Soc. 3, 213 {1972). sb S. B. BARBER and W. F. HAYES, Comp. Bioehem. Physiol. 11, 193 (1964).
