Second Annual Meeting of the European Association for the Study of Diabetes abstracts

EUROPEAN ASSOCIATION FOR THE STUDY OF

DIABETES

A S S O C I A T I O N E U R O P ] ~ E N N E

P O U R L ' ] ~ T U D E

D U D I A B ~ T E

E U R O P A I S C H E

G E S E L L S C H A F T F13R

D I A B E T O L O G I E

ORGANIZATION SECTION

P r e s i d e n t : F . G . Y o ~ x o , Cambr idge Vice -p res iden t s : K. LUNDBAEK, A a r h u s

E. MARTIX, Gen@ve P a s t p r e s i d e n t : J . P . I-IoE~, L o u v a i n

OFFICERS

Secre ta ry : A . E . RENOLD, Gen@ve Treasu re r : E . F . PFEIFFER, F r a n k f u r t Ed i to r - in -ch ie f of Diabe to log ia :

K. OBERDISSE, Diisseldorf

COUNCIL

TER~ EXPIRING 1967 M. JE~SInD, K l a m p e n b o r g �9 R. L ~ T , S t o c k h o l m H. MASKE, W i e n �9 J . L . ROD~mUEz-MIgrox, M a d r i d S. SmNO~ELLI, C a t a n i a

TI~I~IY[ NXPII%ING i 969 P . A . BASTENIE, Bruxel les �9 A. CZYZYK, W a r s z a w a T. 1%. F~ASE~, L o n d o n �9 A. LOUBATI~I~ES, Mont - pel l ier �9 I . PAVEL, B u c a r e s t

H O N O R A R Y M E M B E R S

C.H. BEST, T o r o n t o �9 Sir t{EN~u DALE, L o n d o n �9 H .C . HAGEDO~N, C o p e n h a g e n B . A . Ho~ssAY, B u e n o s Aires �9 R . D . LAwnaENeE, L o n d o n

S U P P O R T I N G M E M B E R S

Ames Europe, Slough, B u c k i n g h a m s h i r e �9 Bayer, L e v e r k u s e n �9 Boehringer, M a n n h e i m �9 Chemie-GriSnenthal, Stolberg �9 Ciba, Base l �9 Carlo Erba, Milano �9 Geigy, Base l �9 Hoechst, F r a n k f u r t �9 Hoffmann-La Roche, Base l �9 Horlick& Windsor , B e r k s h i r e �9 Hormonchemie, Miinchen �9 2Vordisk Insulinlaboratorium, Copenhagen �9 Novo, Copenhagen �9 Organon, Oss-

Pfizer Europe, Bruxel les e t Zi i r ich �9 Sandoz, Base l �9 Schering, Ber l in �9 Vitrum, S t o c k h o l m

Second Annual Meeting of the European Association for the Study of Diabetes

Abstracts

A a r h u s , D e n m a r k , J u l y 6 -8 , 1966

1. Interpretation of the glucose tolerance test. M. ABrAmS, H. KEEN a n d J . F~LLE~. Dep t . of Medicine, G u y ' s Hosp i t a l , London .

The d i s t i nc t i on b e t w e e n a n o r m a l a n d a n a b n o r m a l b lood sugar response to glucose load ing is a t p r e s en t b a s e d u p o n b lood sugar va lues a t a g iven t i m e a f t e r t he load -- com- m o n l y t h e level a t 60 a n d / o r 120 minu te s . D u r i n g t h e glucose to le rance tes t , five or six e s t ima te s of b lood sugar a re usua l ly m a d e ; n o t to use t h e whole of t h i s i n f o r m a t i o n seems prof l igate . W e h a v e the re fo re t r i ed to fi t (by t h e use of a compute r ) curves of va r ious fo rms to our obse rved da ta , in t h e hope of d i s t i ngu i sh ing d i s c r i m ina to ry curve pa r ame te r s . P r e l i m i n a r y analys is ind ica tes t h a t t h i s could be a p romis ing a p p r o a c h to t he d i s t i nc t i on b e t w e e n n o r m a l a n d a b n o r m a l glucose to le rance . Th i s p a p e r gives t he resu l t s of t h e empi r ica l ana lys i s of 574 oral glucose to le rance curves o b t a i n e d f rom a r a n d o m p o p u l a t i o n sample , s t ra t i f ied b.y age a n d sex, a n d seeks to define a b n o r m a l to le rance m a s t a t i s t i c a l l y accep tab le m a n n e r .

2. Vergleichendc Untersuchungen fiber biologische und immunologische Eigenschaften voll- und halbsyntheti- scher Insuline. J . AM~ox , H. DITSC~UNEI~, F. MELANI, E . F . P F E I F F ~ , B. GTJT~E u n d I-I. ZA~N. A b t e i l u n g fiir K l in i sehe E n d o -

kr inologie de r J o h a n n -Wol f gang- G o e t h e - U n i v e r s i t g t F r a n k f u r t a m Ma in u n d Deu t sches W o l l f o r s e h u n g s i n s t i t u t de r T e c h n i s c h e n H o c h s c h u l e Aachen ,

Bei der B e s t i m m u n g de r A k t i v i t g t yon I n s u l i n e n aus na- t i i r l i chen A- u n d B - K e t t e n , aus s y n t h e t i s c h e n A- u n d B- K e t t e n , aus s y n t h e t i s c h e n A - K e t t e n u n d na t f i r l i ehen B- K e t t e n u n d aus na t i i r l i ehen A - K e t t e n u n d s y n t h e t i s e h e n B - K e t t e n muter Z u h i l f e n a h m e biologischer u n d rad io- i m m u n o l o g i s e h e r Me thoden , e rgeben sieh we i tgehende U b e r e i n s t i m m u n g e n . Die Syn these der e inze lnen K e t t e n is t d u r e h A n w e n d u n g der a u t o m a t i s e h e n P e p t i d a n a l y s e n a c h de r y o n Merrif ield en tw icke l t en M e t h o d e wesen t l i eh v e r e i n f a c h t u n d in weft gr6iserem U m f a n g mSglich. D u t c h ve r sch iedene K o m b i n a t i o n s v e r f a h r e n k o n n t e in l e tz te r Zei t die A u s b e u t e bei de r Vere in igung be ide r K e t t e n bis zu 7 0 % ges te iger t werden . E ine m e h r f a c h e R e i n i g u n g de r K o m b i n a t i o n s p r o d u k t e f i ih r t zu k r i s t a l l i s i e rba ren In su - l inen, welche bis zu 100% der A k t i v i t g t y o n n a t i i r l i c h e m Schweine insu l in aufweisen. Versuche , die W i r k u n g der In su l ine a m isol ier ten F e t t g e w e b e der R a t t e d u t c h Ant i - seren y o n Meer schwe inchen b e k a n n t e r K o n z e n t r a t i o n zu h e m m e n zeigten, daiS eine k o m p l e t t e R e a k t i o n m i t den A n t i k S r p e r n erfolgt . Be i d e m r a d i o i m m u n o l o g i s e h e n Ver- f a h r e n e r g a b e n s ieh j edoch A n h a l t s p u n k t e dafi ir , daiS q u a n t i t a t i v un te r seh ied l i ehe R e a k t i o n e n m i t den Ant i -

Vol. 2, No. 2, 1966 Organizat ion Sect ion �9 Abs t rac t s 135

k6rpern auf t re ten, die wahrscheinl ich auf Aggrcgatb i ldun- gen zuri iekzufi ihrcn sind.

Comparative studies on the biological and immunological properties of synthetic and semisynthetic insulins. J. A~Mo~, I-I. DITSC~UNEI% F. I~/IELANI, E.F. PFEIFFER, B. GUTTE and H. ZAHN. Department of Clinical Endo- crinology, Johann-Wolfgang-Goethe University, Frank- furt am Main and the German Wool Research Institute, Institute of Technology, Aachen.

Determinations of the activities -- using bio]ogical and radioimmunological methods -- of insulins synthetized from natural A- and B-chains, from synthetic A-chains and natural B-chains, from natural A-chains and synthetic B-chains and from completely synthetic A- and B-chains, gave a large measure of agreement. The synthesis of the ind iv idual chains is grea t ly simplified by use of the m e t h o d of pept ide analysis developed by 2r and can be carr ied ou t on a larger scale. Several me thods of combina t ion used recent ly have enabled the yield on combining the two chains to be increased to as m u c h as 70%. R e p e a t e d puri f icat ion of the combina t ion p roduc t results in crystal l izable insulins t h a t show up to 100~ of the ac t iv i ty of na tu ra l pork insulin. A t t e m p t s to in- h ibi t the act ion of the insulins on isolated ra t adipose tissue by guinea-pig ant isera in known concent ra t ions showed t h a t the react ion wi th the ant ibodies was complete. Using rad io immunologica l methods , however , there were indicat ions of quan t i t a t i ve differences in the reac- t ions wi th t he ant ibodies , p robab ly due to the format ion of aggregates.

3. Achilles reflex time in diabetic patients. DIM. ANDREEV and D. STRAS]~I~IROV. Diabetes Research Uni t , Bulgar ian A c a d e m y of Sciences, Sofia..

Pho tomyograph ie recording of the Achilles t endon reflex was carr ied ou t in 100 pa t ien ts wi th diabetes mell i tus. I n 28 pa t ien ts there was bi la tera l Achilles t endon areflexia. The m e a n half t ime of re laxa t ion in the remain ing 72 pa- t ients was in the normal range (340 ~: 50 msec), bu t i t was s ignif icantly prolonged when compared wi th a group of 118 normals (314 • 43 msec). I n 59 diabet ics wi thou t clinical signs of neuropa th ic complicat ions the half t ime of re laxa t ion was no t significantly different in compar ison wi th t h a t of the pa t ien ts disclosing clinical diabet ic neuropa thy . I n mos t of the pa t ien ts wi th Achilles t endon areflexia i t was possible to elicit muscle cont rac t ion af ter a di rect b low on the gastroenemius, thus demons t ra t ing tha t in such cases the reason for the areflexia migh t be an impa i rmen t of t he mo to rne rve funct ion. A compar ison is m a d e be tween the dura t ion of Acchilles t endon reflex and the age, du ra t ion of the disease, t ype of diabetes, adequacy of control , t ype of t he rapy and different diabet ic complicat ions. The usefulness of photo- m o t o g r a p h y as a screening m e t h o d for diabetes and early diagnosis of d iabet ic n e u r o p a t h y is stressed.

4. Acute diabetic syndrome in rats, produced with anti- insulin serum from guinea pigs. D. ANDREEV, ST. DI~zov, D. S~RASHIMI~OV and G. DASKEV. Diabetes Research Uni t , Bulgar ian Acad. of Sciences, Sofia.

An acute diabet ic syndrome was p roduced in male Wis ta r rats by means of in t raper i tonea l in ject ion every i2 hr. of ant i - insul in serum f rom guinea pigs. By tes t ing three different doses of ant ibeef- insul in sera (2 ml, 3 ml and 5 ml), i t was observed tha t the 5 ml dose was the mos t effective in producing hyperg lycemia (max. 304 m g % ) ,

glycosuria (manifested early, before the 2nd hr af ter the first inject ion) and ace tonur ia (at the 2nd hr.) I n all exper iments the blood sugar was highest 2 - - 4 hr. af ter the inject ion, decreasing to near the normal at 6 - - 8 hr and increasing sl ightly 2 hr before the nex t inject ion. Some animals were followed for 96 hr and a progressive decrease in the hyperg lycaemic react ion was noted, so t h a t in the last 24 hrs the blood sugar remained in the normal range, despite the in ject ion of the anti- insulin-serum. In the control animals the blood sugar var ied wi th in the normal limits. I n the pancreas of the diabet ic rats an ex t reme degranu- la t ion of the B-cells was found. The la t ter , when compared wi th the controls, failed to fix insulin antibodies, which were labelled wi th fluorescein isothiocynate . These findings demons t ra te the existence of a dis t inct dynamics in the deve lopment of the an t ibody-produced , acute diabet ic syndrome which does no t fully coincide wi th the an t ibody injections. The animals gradual ly be- came resis tent to the diabetogenic act ion of the antisera, and af ter the 6th in ject ion remained normoglycaemic and aglycosuric despite fresh inject ions of ant iserum.

5. Diabetic-like vascular changes in guinea pigs treated with insulin. D. ANDREEV, G. DASIIEV and S~. DI~zov. Diabetes Re- search Uni t , Bulgar ian Acad. of Sciences, Sofia, Bulgaria .

I n order to e laborate highly ac t ive ant i- insul in sera, 50 guinea pigs were immunized wi th insulin in Freunds ' s a d j u v a n t for periods of 3 - - 8 months . All animals produced scra wi th a high immune t i t re (mean 1 : 5000). Af te r bleeding, the kidneys, pancreas, l iver, heart , eye grounds and o ther internal organs were examined. The mos t interest ing da t a are those concerning the k idney glo- merular tufts . The capi l lary basal membrane was found to be th ickened and inf i l t rated wi th a PAS-posi t ive mu- copolysaechar ide substance. This substance formed a t some points small nodules, resembling the nodules seen regular ly in cases wi th diabet ic glomerulosclerosis. The glomerul i were small and wrinkled. The walls of the small vessels a round the capsule were also th ickened ; the lumen of these vessels na r rowed; the inters t i t ia l space inf i l t rated wi th lymphoeytes . The th ickened vessel walls fixed insulin ant ibodies which were labelled wi th fluorescein isothiocyanate . All these changes demons t ra te the existence of an au to immune pathogenesis of the diabet ic vascular changes.

6. Insulin-producing medialstinal tumor, relapsing seven years after operation. D. ANDREEV, G. DASttEV, B. KI~ASTEV and A. BOZlKOV. Diabetes Research Uni t , Bulgar ian Acad. of Sciences, Sofia, Bulgaria.

Recent ly , observat ions of medias t ina l tumors accompanied by severe hypoglycaemie crises have been qui te common. We had the oppor tun i ty to follow such a case for a per iod of 7 years. A 55 year old male had severe hypoglyeaemia crises, associated wi th a large medias t ina l tumor . Af te r opera t ive ex t i rpa t ion of the tumor , which weighed 3200 g, the hypoglycaemic crises ceased and the pa t i en t was in good condi t ion for a per iod of 7 years, at which t ime X - r a y examina t ion revealed a new tumor format ion at the same place. To lbu tamide and big~aanide tolerance tests provoked severe hypoglycaemic crises. The t umor was again r emoved and the crises ceased. The tmnor was composed of undifferent ia ted, par t ia l ly fusiform cells wi th l ight vesicular nuclei. The tissue gave an intensive rest lumi- nescence when t rea ted wi th fluorescein isothiocyanate- labelled ant i- insul in serum f rom guinea pigs. An acid-

136 Organ iza t i on Sect ion �9 A b s t r a c t s Diabetologia

e t h a n o l t i ssue e x t r a c t caused a severe hypog lycaemic shock in r abb i t s . The e x t r a c t was composed of a m i x t u r e of a m o r p h o u s a n d crys ta l l ine insulin. I t is be l i eved t h a t t h e t u m o u r p roduced insu l in or a s t r u c t u r a l l y s imi lar insul in-l ike subs tance .

7. The measurement of the rate of glucose utilization in the red blood cells of diabetics. L. ANGHELESCU a n d G. BACA~U. Medica l Ins t . , Dep t . of N u t r i t i o n , T imisoara , R o u m a n i a .

I n med ica l prac t ice , t he i nves t i ga t i on of t h e me tabo l i c c a r b o h y d r a t e defect in d iabe tes mel l i tus is m a d e b y t he d e t e r m i n a t i o n of b lood sugar a n d t h e e s t i m a t i o n of t he loss of glucose in t he ur ine. The m e t h o d s for t h e de ter - m i n a t i o n of insul in- l ike a c t i v i t y in p l a s m a are too labo- r ious to be c u r r e n t l y used. I n th i s paper , t he e s t i m a t i o n of insul in- l ike a c t i v i t y in p l a s m a was car r ied ou t us ing a s imple m e t h o d based u p o n t he d e t e r m i n a t i o n of t h e r a t e of glucose u t i l i za t ion in e ry th rocy te s . The e ry th rocy te s , in con t ac t w i t h t he i r own p l a s m a in v i t ro a t 37~ consume glucose as a f unc t i on of t h e i r p l a s m a insul in- l ike ac t iv i ty . Th i s asse r t ion is ba sed u p o n t he d e t e r m i n a t i o n of glucose u t i l i za t ion us ing samples f rom 23 persons w i t h o u t me tabo l i c or endocr ine diseases, a n d 25 p a t i e n t s w i t h d iabe tes mel l i tus of differ ing sever i ty . Compared w i t h a m e a n n o r m a l va lue of 4.783 ~: 0.306 /~moles glucose/g H b / h r , t h e r ed b lood cells of d iabe t ics ut i l ize b e t w e e n 0.315 a n d 3.987 #moles /g H b / h r . The a d d i t i o n of insu l in in v i t ro , or in v ivo to d iabet ics , increases t h i s va lue s ignif icant ly . T he m e t h o d can be ut i l ized for t h e e s t i m a t i o n of t h e seve r i ty of t h e disease, a n d fol lowing t r e a t m e n t in d i abe t e s me]l i tus .

8. Cin6tique de l'insuline eristallis@e et de l'insuline radio- iod6e. Y. ARNOULD, F. CANTRAINE, C. DELCItOIX, H . A . OoMs e t J . R . M . FRANCKSON. Lab . de Mdd. exper , e t Lab . cen- t r a l de Mdd. nuclgaire , Un iv . de Bruxel les , Belgique.

On a gtudig les courbes de d i spa r i t i on de l ' insu l ine du p l a s m a apr~s in j ec t ion i n t r ave ineuse r ap ide d ' in su l ine er is ta l l ine (1 & 100 U/kg) et d ' insu l ine m a r q u d e au radio- lode (0.4 k 0.8 mC), de la 2@me ~ la 420@me m i n u t e , sur 25 ehiens n o r m a u x anesthdsids, m a i n t e n u s en normogly - cdmie pa r c o m p e n s a t i o n glucosge. L ' i n su l ine p l a s m a t i q u e a 6t6 dd te rminde p a r dosage r a d i o - i m m u n o l o g i q u e ; la r ad ioae t iv i td insu l in ique a 6td isolde p a r c h r o m a t o g r a p h i c sur papier , i m m u n o c h r o m a t o g r a p h i e e t doub le precipi- t a t i o n des an t icorps . L ' a n a l y s e m a t h g m a t i q u e (effectude au m o y e n d ' u n ordi- n a t e u r I B M 7040) a m o n t r g que ees courbes complexes t r a d u i s a i e n t la s o m m e d ' a u moins t ro is t e r m e s exponen- tiels. E t a n t donn6 ces rdsul ta t s , on do l t r e j e t e r le modgle p r d c d d e m m e n t envisag6 qui supposa i t u n m61ange ex- t race l lu la i re avec per tes i rrgversibles. I1 est propos6 d ' a d o p t e r u n module mami l l a i r e d ' a u mo ins 3 compar t i - m e n t s avee r e t o u r a u p l a s m a de l ' insu l ine r a p i d e m e n t adsorb6e sur les t issus pgr iph6r iques . Cet te hypoth@se concorde avec les g tudes fa i tes in v i t ro , qui m o n t r e n t que la con juga i son de l ' insu l ine avec le muscle isold e t la l ib6ra t ion de ce]ui-ei p a r 61ution son t en r a p p o r t avec sa c o n c e n t r a t i o n dans le mi l ieu d ' i n c u b a t i o n . Elle eat dga- l e m e n t en h a r m o n i c avee des exp6riences prg l iminai res in v ivo fai tes sur des chiens a u d d b u t de la pgr iode qui sui t l ' a b l a t i o n to t a l e d u pancr6as .

Kinetics of crystalline and radioiodinated insulins. Y. AI~NOULD, F. CANTRAINE, C. DELCROIX, H . A . 0oMs a n d J.I%.M. FRANCKSOX. L a b o r a t o r y of E x p e r i m e n t a l Medicine a n d Cent ra l L a b o r a t o r y of Nuc lea r Medicine, U n i v e r s i t y of Brussels , Belg ium.

P l a s m a d i s a p p e a r a n c e curves r ecorded fol lowing r ap id i n t r a v e n o u s in jec t ion of c rys ta l l ine insu l in (I to 100 U/kg) a n d r a d i o i o d i n a t e d insul ins (0.4 to 0.8 mC) were s tud ied f rom t h e 2nd up to t h e 420 th . m i n u t e in 25 h e a l t h y a n a e s t h e t i z e d dogs m a i n t a i n e d in n o r m o g l y e a e m i a b y glucose compensa t ion . P l a s m a insu l in was m e a s u r e d b y r a d i o i m m u n o a s s a y ; insul inic r a d i o a c t i v i t y was i so la ted b y p a p e r c h r o m a t o g r a p h y , i m m u n o c h r o m a t o g r a p h y a n d doub le a n t i b o d y p rec ip i t a t ion . M a t h e m a t i c a l ana lyses pe r fo rmed w i t h a digi ta l c o m p u t e r I B M 7040 revea led t h a t these complex curves were b e t t e r expressed as t h e sum of a t leas t 3 exponen t i a l t e rms , a n d consequen t l y t h e mode l p rev ious ly assumed, wh ich compr i sed a n ex t race l lu la r m i x i n g pool w i t h i r revers ib le losses, m u s t be re jec ted . A m a m i l l a r y mode l of a t l eas t 3 com- p a r t m e n t s w i t h f eedback to p l a s m a of insu l in in i t i a l ly a d s o r b e d to pe r iphe ra l t i ssues is suggested. This hypo- thes is is in concordance w i t h work in v i t ro showing t h a t b i n d i n g of insu l in to, a n d i ts e lu t ion f rom iso la ted muscle , are relevant to its concentration in the incubation medium. It also agrees with preliminary experiments in vivo, carried out in dogs during the early stage of total pan- createctomy.

9. Dosage radio-immunologique du glucagon plasmatique chez l ' h o m m e . 1~. ASSAN, G. ROSS]~LIN, G. TCHOBROUTSKY, J . D~OTJET, J . DOLAIS et M. DEROT. HSte l -Dieu , Par is , F rance .

L a d d t e r m i n a t i o n r a d i o - i m m u n o l o g i q u e du t a u x p lasma- t ique du g lueagon ehez l ' h o m m e a 6td effectude ~ l ' a ide d ' u n sys tbme e o m p o r t a n t : u n s t a n d a r d de g lucagon de pore (Lilly, recristal l isd 2 lois, lot n ~ S -- 8964), u n an t i - sd rum o b t e n u chez le l ap in selon u n procddd prde6dem- m e n t ddcr i t e t du g lucagon de pore iod6 selon la t echn i - que u t i l i s an t la ch lo ramine T avee une ac t iv i td spdeifique m o y e n n e de 400 #C//~g. L a sdpa ra t ion en t re g lucagon l ibre et g lucagon lid ~ l ' an t i co rps es t fa i te pa r c h r o m a t o - d lect rophorbse sur pap ie r W h a t m a n n 3 MM. Le sys t~me est sensible & l ' a d j o n c t i o n de 50 /~#g/ml de glucagon. 80 p rd lbvemen t s on t dt6 6tudids chez 18 ma lades a u t ou r s d ' dp reuves dynamiques . Les t a u x k j eun de glu- eagondmie son t eompr i s en t r e 0.4 e t 1.5 m/~g/ml ehez 4 su je ts n o r m a u x . Apr@s a d m i n i s t r a t i o n de 50 gr de glucose p a r m 2 de surface corporelle, la g lucagondmie s 'dlbve m o d d r d m e n t chez les n o r m a u x e o m m e chez les d iabd t iques de 0.5 & 2 m ~ g / m l pou r a t t e i n d r e en 30 m i n u t e s des va leurs compr ises en t r e 2 e t 2.5 m/~g/ml, puis r e t o u r n e aux va leurs no rma le s en t r e la 60bme et la 120brae m i n u t e . El le r e m o n t e s e c o n d a i r e m e n t en t r e 120 e t 240 minu te s . Chez u n su je t p r d s e n t a n t une hypoglycdmie fone t ionne l le rdact ive , l ' i n j ee t i on I .V. de 1 gr de t o l b u t a m i d e a en t r a ind une dldvat ion tr@s f r anche de la g lueagondmie de 0.4 re#g/ ml ~ 5 m#g /ml , alors que chez ce m@me suje t , apr~s ad- m i n i s t r a t i o n de glucose p a r vole orale la g lucagongmie es t restde k 0.5 m ~ g / m l p e n d a n t la durde de l 'dpreuve. Chez u n su je t p r d s e n t a n t une hypog lycdmie o rgan ique p a r addnoea re inome l a n g e r h a n s i e n la g lucagondmie & jeun est dlevde: 2.8 m~g/ml . Le t a u x s ' aba isse au eours de l ' 6preuve d ' hype rg lyedmie p rovoqude p a r vole orale. Chez u n a u t r e m a l a d e a t t e i n t d ' hypog lyedmie o rgan ique pa r addnome l ange rhans i en , la g lueagondmie le p remie r jour d ' u n e dpreuve de r e s t r i c t ion ealor ique et g luc id ique a dtd t rouvde & 12, 9, 4 et 2 m/~g/ml & des m o m e n t s diffdrents de la mgme journde en prdsenee d ' u n t a u x ba s de gly- edmie et de va leurs dlevdes d ' insu l indmie et d ' t i . G . I t . A u eours de 3 dpreuves e o m p o r t a n t des per fus ions de 25 gr d ' a r g i n i n e p ra t iqudes ehez des su je ts n o r m a u x ou diabd- t iques , il a dtd t r o u v d une d ldvat ion moddrde ma i s n e t t e de la g lueagondmie en t r e la 30bme et la 60bme m i n u t e (1.5 g 3 m/~g/ml). Cet te d ldvat ion n ' a pas dtd r e t rouvde dans les m@mes ddlais au cours de t e s t s g l ' insul ine. L a conf i rma t ion des effets d u t o l b u t a m i d e e t de l ' a rg in ine

Vol. 2, No. 2, 1966 Organization Section �9 Abstracts 137

sur la s6cr6tion du glucagon font l 'objet des 6tudes ac- tuelles.

Radio-immunological determination of plasma glucagon in man. i%. ASSAN, G. ]~OSSELI~, G. TC~O~OUTSKY, J. D~OUE~, J. DOLAIS and M. DE~o~. HStel-Dieu; Paris.

Radio-immunological determination of the plasma level of glucagon in man was carried out with the following materials: a standard preparation of hog glucagon (Lilly, recrystallized twice, batch no. S-8964); an antiserum obtained from rabbits by a previously described method; and hog glucagon iodinated according to the technique employing chloramine-T with a mean specific act ivi ty of 400 #C/#g. The separation of free glucagon and antibody- linked glucagon was obtained by chromato-electrophoresis on Whatman 3 MM paper. This system is sensitive to the addition of 50/~#g/ml of glucagon. Eighty samples from 18 patients were studied over a period. In four normal subjects the fasting glucagon levels varied between 0.4 and 1.5 m/~g/ml. After administration of 50 g glucose per m 2 body surface, the glucagon levels rose to a moderate degree in both the normal and the diabetic subjects from 0.5 to 2 m#g/ml to reach in 30 minutes values between 2 and 2.5 m/~g/ml; then they returned to normal levels between the 60th and 120th minutes. There was a secondary rise between the 120th and 240th minutes. In a subject with functional hypoglycaemia intravenous injection of I g tolbutamide caused a marked rise in the glucagon levels from 0.4 m/~g/ml to 5 m#g/ml, whereas after oral administration of glucose the levels remained at 0.5 m#g/ml during the period of the test. In a patient with hypoglycaemia caused by an islet-cell adenocarcinoma of the pancreas the glucagon fasting levels were high: 2.8 m#g/ml. The levels fell during the oral glucose tolerance test. In another patient with hypoglycaemia caused by islet-cell adenoma the glucagon levels on the first day of a test restricting his calorie and carbohydrate intake were 12, 9, 4 and 2 m#g/ml at different times, while the blood sugar level was low and the insulin and HGH levels were high. In three tests involving the perfusion of normal and diabetic subjects with 25 g arginine, a moderate but distinct rise in the glucagon level (1.5 to 3 m#/g/ml) was found to occur between the 30th and 60th minutes. No rise was found at these times in tests with insulin. Studies are in progress to confirm the effects of tolbutamide and arginine on glucagon secretion.

10. Glycodiazine (Lycanol) in the management of ma- turity onset diabetes. H. AYAD. Endocrine and Metabolic Unit, E1-KhMig Ge- neral Government t{osp. Cairo, Egypt.

Lyeanol is a glycodiazine which differs from the sul- phonylurea group of oral hypoglycemic compounds in being a sulphapyrimidine derivative. The drug was used in the management of cases of diabetes mellitus, of the matur i ty onset type, during the last eighteen months and experience with hundred cases will be dicussed. The outcome of therapy was found to be closely related to the insulin needs of the individual before institution of the drug. The individuals whose previous insulin re- quirements were 40 units or less daily, showed excellent and prompt control, as a whole. In patients whose insulin needs exceeded 60 units daily, the result of therapy with Lycanol was poor. A good percentage of these cases could, however, at tain good control when a biguanide (Metformin, Glucophage) was combined with Lycano]. The most remarkable effect noticed was the effect of Lycanol on postprandial hyperglycemia, which it did control in the majori ty of cases.

Unlike the tolbutamide group of drugs, which, though lowering the fasting hyperglycemia satisafactorily, usually fail to effect postprandiM hyperglycemia, Lycanol lowers and effectively controls both fasting and postprandial hyperglycemia. This property of the drug may be ascribed either to its rapid absorption from the gut and thus the early onset of its pharmacological effect, or to the fact that Lycanol has more potent insulin releasing properties. No toxic or inconvenient side effects were noticed during Lycanol therapy. I t is concluded that Lycanol is a medinm-acting, oral hypoglycemic agent, and is a safe drug in the long-term management of cases of matur i ty onset diabetes mellitus.

11. Le d4pistage du diab6te sucr6 chez les mMades ~ infarctus myocardique. G. BACANU et L. ANGItELESCU. Clinique m6dicale N ~ 1 Section Nutrition-Diab6te, Timisoare, Roumanie.

L'infarctus myocardique peut constituer ]e moment favorable pour d6pister l 'existence d 'un diab6te sucr6. Certains cardiologues at tr ibuent l 'hyperglyc6mie et la glycosurie h l 'infarctus, tout en ]es consid6rant transi- toires. Parfois on n'ex6cute m6me plus de pareilles investigations. Nous avons fair des 6tudes chez 55 malades dont les ant6c6dents personnels pr6sentaient un infarctus myocardique. Ces malades ignoraient qu'ils avaient un diab6te sucr6. Nous avons d6pist6, par la glyc6mie k jeun et par la glyeosurie, un diab6te sucr6 manifeste chez 45% d'entre eux, et par double surcharge en glucose per os, un diab6te latent chez 19 %. Dens ce dernier groupe, 21% des malades mentionnaient l 'existence d 'un diab6te dans leur famille. Nos r6sultats concordent avee ceux de B~3- CHELE, FREHNER, GLIGOttE et GOLDBElCGEE. La r6alisa- tion, dens un d61ai de 5 ~ 6 semaines apr6s ]'infarctus, de l 'une des 6preuves de d6pistage du diab6te sucr6, manifeste ou latent, devrait 6tre obligatoire pour cheque malade. On 6viterait ainsi que les malades ayant eu un infarctus myocardique et pr6sentant un diab6te ignor6, re- 9oivent un r6gime alimentaire hyperglucidique pour fa- voriser la convalescence. De cette mani6re, on 6viterait la d6compensation d 'un diab6te non d6pist6 et ]a progression des complications vasculaires concomitantes.

The detection of diabetes mellitus in patients with myocardial infarction. G. BACANU and L. ANGHELESCU. Medical Clinic, No. 1 Section (Nutrition and Diabetes), Timisoara, Romania.

A myocardial infarction may provide an opportunity for detecting diabetes mellitus. Some cardiologists attribute hyperglycaemia and glycosuria to infarction, while re- garding them as transient. Sometimes the tests are not even carried out. The authors investigated 55 patients with a past history of myocardial infarction. These patients did not know whether they had diabetes mellitus or not. By deter- ruination of their fasting blood sugar and tests for gly- eosuria, overt diabetes was detected in 45%, and by an oral glucose tolerance with a double loading, latent diabetes was detected in a further 19%. Among the latter, 21% had a family history of diabetes. Our findings are in agreement with those of Bi~CHELE, FREIINER, GLIGOI~E, GOLI)BEI~GEI~ and others. Every patient who has had a myocardial infarction should, within a period of 5-- 6 weeks after its occurrence, be subjected to one or other of the tests for the detection of overt or latent diabetes. This will prevent patients who have had an infarct and have undetected diabetes from being placed on a diet too rich in carbohydrates, intended to provide them with the necessary calories. In this way decompensation of the diabetes will be avoided, as well

138 Organization Section. Abstracts Diabetologia

as the progression of the accompanying vascular complications.

12. Influence of sodium-~-hydroxybutyrate on glucose and free fatty acid metabolism in normal dogs. E. BALASSE, IK.-A. OOMS, Y. AR~COULD and V. COWARD. Dept. of General Pathology and Exptl. Med., University of Brussels, Belgium.

The metabolic effects of a 120 minutes infusion of Sodium- fl-hydroxybutyrate (5 mM/Kg/h) were studied in 35 healthy anaesthetized dogs. Arterial blood sugar decreased progressively, the fall averaging 20 rag/100 ml at the end of infusion. A slight, transient increase in plasma insulin, (measured by radio- immunoassay), was observed, with a mean peak of + 7 /~U/inl at the tenth minute of infusion. The disappearance rates of a 12C-glucose load and a tracer of 14C- glucose were not modified by the sodinm-fi-hydroxybutyrate infusion. Isotope dilution calculation showed that hypoglycemia was entirely the consequence of a reduction in hepatic glucose output. Decrease in plasma level of free fatty acids averaged 200 #Eq/l. Turnover studies of laC palmitate, using the constant infusion technique, revealed that the inhibition of FFA release by adipose tissue was the major phenomenon. Contrary to previous work, our data do not suggest that the prolonged inhibitory effect of sodium-/~-hydroxybutyrate on liver glucose output could be mediated through the minor pancreatfc stimulation. However, the insulin changes could explain the FFA changes observed.

13. Acides gras libres Plasmatiques et M@tabolisme des Glucides chez l'homme normal. P. BARJON, A. CI~,ASTES DE PAULET et B. DESCO~PS Chaire de Pathologie mddicale, Laboratoire de Biochimie, Facult6 de Mddeeine, Montpellier. L'6tude des eorrglations du mgtabolisme des A.G.L.P. et du glucose, amorcde en 1956 par le travail original de Dole, a abouti ~ la notion de cycle physiologique glucose/ A.G., developpg par Randle depuis 1963; ainsi s'explique l'intdr@t port6 aux @reuves dynamiques qui sollieitent les dl@ments constitutifs de ce cycle. L 'un des deux aspects de ee cycle a fair l 'objet de ce travail: les modi- fications des A.G.L.P. ont dtg recherchdes au eours d'@reuves modifiant la glycdmie (injection I.V. de glucose, de glucagon, de tolbutamide et d'insuline et injection S.C. d'insuline). La vitesse de diminution des A.G.L.P. sous l'effet du glucose I .V . (0.3 mg/kg) peut s'exprimer par un coefficient lid au coefficient d'asslmilation glueidique par une relation lin4aire; par eontre, nous n 'avons pu retrouver de corr@lation entre KG et le pourcentage de diminution des A.G.L.P. mesurd ~ un temps donnd. Nous avons retrouv@, comme Lipsett et Lefebvre Faction diphasique du glucagon (3 mg I.V.) sur les A.G.L.P: la phase initiale de diminution parait secondaire k l'hyperglycdmie et la rgascenclon traduit l'effet lipolytique de eette hormone, mis en 6vidence in vitro. L'@reuve ~ l'insuline I .V . (0.1 U/kg) est d 'un int6r@t m@diocre, en raison des effets adrdnergiques r@actionnels k l'hypoglyc@mie brutale. L'insuline S.C. (0.2 U/kg) provoque une diminution nette et prolong6e des A.G.L.P. Le Tolbutamide (1 g I.V.) a des effets comparables b~ l ' insuline S.C. Au eours de ees diff@rentes 6preuves, l 'analyse des A.G.- L.P. par chromatographic gaz-liquide n'a montr6 aucune variation significative du pourcentage de chaque A.G. : cette eonstatation sugg@re que la mobilisation des A.G. des lipides de r@serve s'effectue de fa~on uniforme. Ces @preuves sont suseeptibles d'aider ~ mieux eomprendre lee relations physiopathologiques du mgtabolisme des glucides avec le mdtabolisme des lipides chez le diabdtique.

Free fatty acids and carbohydrate metabolism in normal subjects. P. BAI~JON, A. CI~ASTES DE PAUV.ET and B. DEscoMPs. Pathology Department and Biochemical Laboratory, Fa- culty of Medicine; Montpellier.

The study of the correlations between the metabolism of plasma free fatty acids and of glucose, initiated by Dole's original work in 1956, has led to the concept of a physiological glucose-fatty-acid cycle. This concept has been developed by l~andle since 1963. It explains the interest taken in tests to elucidate the dynamics and constitutional elements of this cycle. One of the two aspects of this cycle was the subject of this study: to bring about changes in the free fatty acids of the plasma by means that alter the blood sugar level (intravenous injection of glucose, glucagon, tolbutamide and insulin, and subcutaneous injection of insulin). The rate of fall of the plasma free fatty acids under the influence of intravenous glucose (0.3 mg/kg) may be expressed by a coefficient, which is lineally related to the coefficient of carbohydrate assimilation. On the other hand, we could find no correlation between Ka and the percentage fall of the free fatty acids measured at a given time. Like Lipsett and Lefebvre, we found that glucagon (3 mg I.V.) had a biphasie action on the free fatty acids: an initial phase of fall appeared to be secondary to a rise of blood sugar, and the subsequent rise of free fatty acids indicated the lipolytie effect of this hormone, which had been demonstrated in vitro. The test with intravenous insulin (0.1 U/kg) was of little interest, because of the adrenergic reaction to the sharp fall in the blood sugar. Subcutaneous insulin (0.2 U/kg) caused a marked and prolonged fall in the free fat ty acids. Intravenous tolbutamide (lg) had effects comparable with those of subcutaneous insulin. Analysis by gas-liquid chromatography of the free fat ty acids during the course of these tests showed no significant variation in the proportion of each fat ty acid. This finding suggests that the mobilization of the fatty acids from the reserve lipids is effected in a uniform way. These tests may help in the understanding of the physiopathological relationships between carbohydrate metabolism and lipid metabolism in the diabetic.

14. Eigenheiten des bei Azidose auftretenden Insulin- sehocks. L. BA~TA. I. Kinderklinik der Medizinisehen Universitiit Budapest.

Bei Kaninchen wurde durch viermalige, viertelstiindliche i.v. Verabreichung yon 20 ml einer 40 bzw. 50~oigen Glu- cosel6sung eine Azidose herbeigefiihrt. I n der Mehrheit der F/~lle sank das Blut-pI-I urn 0.1--0.2 ab. Von 23 Ka- ninehen erhielten 13 zusammen mit der letzten Glucose- dosis 64 E. Insulin i .v. ; 10 Tiere erhielten kein Insulin. Die 10 nicht mit Insulin behandelten Tiere blieben am Leben, wogegen 6 yon den insulinbehandelten 13 Tieren 3--4 Stunden naeh der Insulinbehandlung unter Schock- symptomen verendeten. Bei den verendeten Xaninehen war unmit telbar naeh der Insulingabe ein sig~ifikanter Anstieg des Blutzuckers zu beobachten, dem eine Blut- zuckerabnahme folgte; bei den Tieren, bei denen die Symptome des Insulinschoeks nicht auftraten, sank das Blutzueker sehneller und auf tiefere Werte ab, als bei den unter Schocksymptomen verendeten. Das pH und das Standard-Biearbonat fielen bei den insulinbehandelten und verendeten Tieren signifikant schneller ab, als bei den am Leben gebliebenen. Bei den verendeten Tieren werden sowohl dureh den Ablauf der Blutzuckerkurve wie die Ver/~nderung des pH die gegenregulatorischen Wirkungen in den Vordergrund gestellt. -- Verfasser hat in seinen friiheren Untersuehungen die Insulinwirkung bei Azetessigsaurevergiftung gepriift und die Ergebnisse

VoL 2, No. 2, 1966 Organization Section - Abstracts 139

mit den Befunden vergliehen, die bei Pat ienten im auf normo- bzw. hyperglykamischem Niveau auftretenden Insulinschoek erhoben wurden.

Characteristics of insulin shock during acidosis. L. B ~ A . First Paediatrie Clinic of the Medical Univer- sity of Budapest.

Acidosis in rabbits was provoked by four intravenous injections, at 15-minute intervals, of 20 ml of a 40 or 50% solution of glucose. In most cases, there was a fall of 0.1--0.2 in the p]:I of the blood. Of 23 rabbits, 13 were given 64 I U of insulin intravenously together with the last dose of glucose, while the remaining 10 received no insulin treatment. These 10 animals remained alive, whereas 6 of tile 13 insulin-treated animals died 3- -4 hours after the insulin injection, exhibiting shock symptoms. In the rabbits tha t died, the blood sugar rose significantly immediately after the administration of insulin and then fell again; however, in the animals tha t did not develop symptoms of insulin shock the blood sugar fell much more rapidly and to a much lower level than in those that died with shock symptoms. In the latter animals, the pH of the blood and the bicarbonate fell much more steeply than in the animals that remained alive. In the animals that died, both the blood sugar curve and the pH behaved in a manner indicating the predominance of counterregulatory mechanisms. In ear- Her studies, the author has examined the effect of insulin in animals with acetoacetic acid poisoning and compared the findings with observations in normog]ycaemic and hyperglycaemic patients who develop insulin shock.

15. Zur Wirkung yon Buformin auf das Fettgewebe. 1%IJ-DIGER BECK~•162 Biochemische Abteilung der Che- mic Griinenthal GmbH., Stolberg/1%hld.

Die dutch Biguanide bewirkte Blutzuckersenkung beruht vorwiegend auf einer vermehrten Verwert~mg yon Glu- cose im Muskelgewebe (1, 2). Dagegen ist bisher noch ungeklart, ob diese oralen Antidiabetika den Stoffwechsel des Fettgewebes beeinflussen. Entsprechende in vitro- Versuehe ffihrten in Abhangigkeit yon den verwendeten, racist sehr hohen I~ionzentrationen, zu widersprfiehlichen Ergebnissen (3). Wit untersuchten die Wirktmg yon Bu- formin (1-Butyl-gibuanid-hydroehlorid) auf das epididy- male 1%attenfettgewebe mit I(onzentrationen, die den therapeutisehen ]Blutspiegeln entsprechen. Unsere Ergebnisse zeigen, daI3 0.5, 1.0 und 5.0/~g Bufor- min/ml ebenso wie l~henformin (4) die spontane Lipolyse und die dureh 0.2 /~g/ml Noradrenalin aktivierte Fett- spaltung nieht beeinflussen. Weiterhin konnten wit mit Glucose-l-laC und Glucose-U-14C keinen Effekt yon Bu- formin (I /~g/ml) auf die Glucoseaufnahme, die Glucose- oxydation und den Einbau yon Glucose in Triglyeerid- Glycerin und Triglycerid-Fettsauren beobaehten. Die Wirkung yon Insulin (i und i0 mE/m]) auf den Stoff- weehsel des Fettgewebes wird dureh Buformin (i /~g/ml) weder gehemmt noeh potenziert. Literatur : 1. BUTTERFIELD, W. J . H., U. M. J . WHICHELOW : Diabetes 11, 281 (1962). 2. SC~LESS, G.L.: Metabolism 13, 934 (1964). 3. Literaturfibersicht: BECK~ANlV, 1%.: Dtsch. reed. Wsehr. 90, 1589 (1965). 4. S~O~E, D.B., u. J .D . BROWN: 2nd international symposium on drugs af- fecting lipid metabolism, Mailand 1 3 . - 15.9. 1965.

The action of buformin on adipose tissue. 1%. B~CKMAN~. Biochemische Abteilung der Chemie Grfi- nenthal GMBH., Stolberg/1%hld.

The fall in blood sugar brought about by biguanides is due primarily to an increased utilization of glucose by muscle tissues (BUTTERFIELD and W]~Ic~T,ow, 1962; SCHLnss, 1964). On the other hand, it is not certain

whether these oral antidiabeties influence the metabolism of adipose tissue. Experiments in vitro have yielded con- flieting results, depending on the concentration used, which has usually been very high (BEcK~rA~, 1965). We investigated the effect of Buformin (1-butylbiguanide hydrochloride) on rat epididymal adipose tissue at con- eentrations corresponding to those attained in blood when the drug is used therapeutically. Our results show that concentrations of 0.5, 1.0 and 5.0 #g of Buformin per ml and similar concentrations of Phen- formin (S~oNE and B~owlv, 1965) have no influence either on spontaneous lipolysis or on lipolysis induced by 0.2 #g of noradrenaline per ml. Furthermore, using 1-1aC - and U-14C-labelled glucose we were unable to detect any effect of Buformin (1 /~g/ml) on glucose uptake, glucose oxidation or the incorporation of glucose-carbon into triglyeeride glycerol and triglyceride fat ty acids. The effect of insulin (1 and 10 mU/ml) on the metabolism of adipose tissue was neither inhibited nor potentiated by Buformin (1 #g/m1).

16. Einfaehe semiquantRative Bestimmung der Glucose in der Konjunktivalfliissigkeit zur raschen Erfassung starker Hyperglykaemien. W. BE~aGE~. Diabetikerberatungsstelle der Med. Univer- sitatspoliklinil~, Zfirich und Med. Abteilung des Kranken- haus Neumfinster, Zollikerberg.

Bei tiber 200 Diabetikern und Niehtdiabetikern wurde die Glucose in der Konjunktivalflfissigkeit semiquanti tat iv mit Hilfe eines Enzymteststreifens (Tes-Tape, Glucotest), in einzelnen Fallen quanti tat iv mit der Oxydasemethode best immt und mit den gleichzeitig ermitte]ten Blutzueker- werten verglichen..Methode: Ein ca. 4 em langer Test- streifen wird solange mit einem ]Ende in den unteren Konjunktivalsack eingelegt, bis er auf einer Lange yon 0.5--1 cm angefeuchtet ist (1. Teststreifenreaktion (1. T.R.)). Nach 30--60 Sekunden legt man einen zweiten Teststreifen ein (2. T.1%.). (Die 2. T.1%. ist eher positiv als die 1. T. 1%., da der Glucosegehalt im Reizsekret h6her ist als in der basalen Konjunktivalflfissigkeit.) Das er- haltene 1%esultat wird folgendermai3en angegeben: 0 ---- keine Verfarbung, (+) = vereinzelte Stellen hellgrfin, + = homogene bla2griine Verfarbung, + + = he]lgrfine Verfgrbung, + + + = dunkelgriine Verfarbung. Wir erhielten folgende Beziehung zwisehen Blutzucker (BZ) und Teststreifenreaktion: BZ unter 100 m g % 1. T. 1%. immer neg., 2. T.1%. nur vereinzelt (+) , BZ 400--500 rag% 1. T.1%. in 70% + , vereinzelt + + . BZ 600--700 rag% 1. T.1%. immer + + od. + + + . BZ fiber 700 rag% 1. T.1%. immer + + + . In anderen Worten 1. T.R. 0, 2. T.1%. 0 - - ( + ) BZ unter 200 rag%, 1. T.R. + od. + + BZ mei- stens fiber 400 rag%. 1. T.I~. + + + BZ fiber 700 rag%. Quantitative Bestimmungen des Glucosegehalts in der Konjunktivalfifissigkeit ergaben bei BZ-Werten yon 70-- 280 mg% Werte yon 17--68 m g % und bei BZ-Werten yon 600--800 rag% Werte yon 90--120 rag%. Mit dieser Methode sollte es mSglich sein, am Krankenbet t raseh und ohne Blutentnahme eine Normo- oder Hypoglykae- mie yon einer schweren I-Iyperglykaemie zu unterschei- den, was fiir die Differentialdiagnose komatSser Zustande beim Diabetiker yon Nutzen sein kaml.

Rapid detection of severe hyperglycaemia by a simple semiquantitative determination of glucose in the conjunctival fluid. ~r BERGER. Diabetes Advice Centre of the Med. Poll- din., Univ. of Zurich; Med. Dept. of Neumfinster t tosp. Zollikerberg, Switzerland.

In more than 200 diabetics and non-diabetics, the glucose content of the conjunctival fluid was determined semi- quantitatively by means of an enzymeimpregnant test strip (Tes-Tape, Glucotest); in a few cases it was also

Diabetologia, u 2 10

140 Organization section �9 Abstracts Diabetologia

determined quantitatively by the glucose-oxidase method and the results compared with the blood sugar levels determined at the same time. Method: a test strip about 4 cm long was placed with one end in the lower conjunc- tivaI sac and left there unt i l it was moistenend for a length of 0.5--1 cm (lst test strip reaction (1. T.R.)). After 30--60 seconds a second test strip was treated similarly (2. T.R.) (The 2. T.R. is more likely to be positive than the 1. T.1%. because the glucose content of the secretion following irritation is higher than that of conjunctival fluid under basal conditions.). The result obtained was expressed as follows: 0 = no color, (+ ) ~ isolated pale green patches, + = homogeneous pale green color, @ + = green color, + + + ~ dark green color. We obtained the following relationship between the blood sugar (BS) and the test tape reaction: 13S less than 100 rag%, 1. T.R. always negative, 2. T.1%. only occasionally ( + ) ; BS 400-- 500 rag%, 1. T.R. in 70% + , occasionally + + ; BS 600-- 700 rag%, 1. T.R. always + + or + + + ; BS more than 700 mg%, I. T.1%. always + + +. In other words, I. T.I~. 0, 2. T.1%. 0 or (+) = BS less than 200 rag%; I. T.I%. between -]- and + Jr = BS usually over 400 rag%; i. T.1%. ++ + = BS over 700 rag%. Quantitative determinations of the glucose content of the conjunctival fluid gave values of 17--68% for BS levels of 70--280 rag% and values of 90--200 rag% for BS levels of 600--800 rag%. With this method it should be possible at the bedside and without taking a blood sample to make a rapid differentiation between normoglycaemia or hypoglycaemia and severe hyperglyeaemia, which could be of value in the differential diagnosis of comatose states in diabetics.

17. Nachweis der Insulinaktiviti~t im Lebergewebe. A. BEttlNGER, G. GEYEtt, H. THALER, K.H. TRAGL und W. WALD~XUSL. I. Medizinische Universitatsklinik, Wien. An hungernden Kaninchen haben wir beobachtet, daB das Glucagon nur bei gesunden und nicht bei alloxandiabeti- schen Versuchstieren einen Leberglykogenschwund er- zeugt. Erst nach einer gleichzeitigen Injekt ion yon Glu- cagon, und einer stoffwechselaktiven Insulinmenge ge- winnt die alloxandiabetische Leber die Fahigkeit, das Gly- kogen abzubauen. Der Leberglykogenschwund ist abet nicht Ms der Ausdruck einer Insulinwirkung zu werten, da das Insulin ohne Glucagonzusatz bei gesunden und diabetischen Kaninchen eine Leberglykogenzunahme er- zeugt. Dieses Ergebnis 1M3t also erkennen, dab die An- wesenheit yon stoffwechselaktivem Insulin zur normalen Entfal tung des Glucagoneffektes erfordcrlich ist. Ein feb- lender Leberglykogenschwund im AnschluB an eine In- jektion yon Glucagon laBt daher auf einen Mangel an stoffwechselaktivem Insulin schlieBen. Quantitative Be- st immungen des Glykogengchaltes im Leberpunktat yon Altersdiabetikern haben folgendes gezeig~: ])as Glucagon bewirkt bei hungernden Altersdiabetikern keinen Leber- glykogenschwund. Eine gleichzeitige Verabreichung yon Insulin oder Kohlehydraten oder Zuckerbfldern oder oralen Antidiabetika fiihrt zu einem Abbau des Leberglyko- gens, also zu einer Zunahmc der Insulinaktivits im Le- bergewebe. An t Iand weiterer Untersuchungen wird dar- gestelit, wie oft am Tag wiihrend einer I)iatbehandltmg und einer oralen Diabetestherapie aktives Insulin in den Stoffwechsel der Leber eingreift und welchen EinfluB die Insulinantagonisten auf die Insulinaktivit~it in der Leber nehmen.

Determination of insulin activity in liver tissues. A. BERINGE:a, G. GEYEI% H. TH~ER, K.H. TRAOL und W. WALnHs First medical clinic, University of Vienna. We have observed that the administration of glucagon to fasting rabbits caused a disappearance of glycogen from the liver only in healthy animals and not in animals with alloxan diabetes. I n the diabetic animals the ability of

the liver to break down glycogen was restored only after the simultaneous injection of glucagon and a metabolically active dose of insulin. The disappearance of glycogen from the liver is not at tr ibutable to the effect of the insulin, however, since without the addition of glucagon insulin causes an accumulation of glycogen in the liver, both in healthy and in diabetics rabbits. From this result it follows that the presence of metabolically active insulin is necessary for normal glucagon activity. Conse- quently, if an injection of glucagon is not followed by disappearance of glycogen from the liver, this is evidence of a deficiency of metabolically active insulin. Quanti- tative determinations of the glycogen content of liver- puncture specimens obtained from elderly diabetics have shown the following: In fasting elderly diabetics, administration of glucagon causes no disappearance of glycogen from the liver. The administration of insulin, carbohydrates, sugar precursors or oral antidiabetic substances at the same time as the glucagon leads to breakdown of liver glycogen, indicating an increase in the insulin activity in the liver. Evidence from other studies is presented, showing that in patients undergoing dietetic t reatment or therapy with oral antidiabetics, active insulin participates several times a day in liver metabolism and that insulin antagonist may exert an effect on the insulin activity in the liver.

18. Effect of trishydroximethilaminomethane on glucose uptake by epididymal fat pad incubated in vitro in Ringer medium and in human serum (normal and diabetic). M. BIANCHESSI, R. MILA2Vl et C. GALLI. Ist i tut0 di Se- meiotiea Medica, Milan, Italic.

Trishydroximethflaminomethane (TItAM) is used as a buffering substance in the t reatment of diabetic acidosis (1). Results from electrophoresis suggested that it releases globulin-bound insulin in serum of diabetic patients (1); moreover an hypoglycaemie activity of TttAM was shown in normal but not in pancreatectomized dogs, suggesting a stimulation of insulin secretion (2). As was to be expected from these data we found in experiments performed in triplicate in Krebs-1%inger-biearbonate buffer (glucose 300 rag%; gas phase O 3 + COs 5%) that Tt tAM (1, 10, 25, 50 raM) did not stimulate glucose uptake by rat epididymal fat pad either in the absence or in the presence of crystalline bovine insulin {200 mmU/ml + gelatin 200 rag%). ~rhen used at 50 mM concentration, TFIAM did not modify glucose uptake by fat pad incubated in non-diluted serum (reconstituted to a glucose concentration of 300 rag%) of two normal and of two diabetic insulin-requiring subjects. These data seem to exclude that THAM enhances glucose metabolism of adipose *issue (Ringer experiments). The lack of any i~- crease in "total" insulin-like activity (serum experiments) in the presence of THAM, despite the fact tha t it has been described that THAM releases globulin-bound insulin, may be justified by the fact tha t released insulin does not increase "total" insulin-like activity as measured by adipose tissue method (Antoniades 1961). According to this point of view the immunological and the diaphragm muscle assays -- which measure "free" insulin only -- can be expected to give information whether increase of "free" insulin really follows the described releasing of "bound" insulin (i). Bibliography: 1. REES, S.B., et al.: Ann. N.Y. Acad. Sci. 92, 755 (1961). 2. BENNET, T.E., and TA~AIL: Ann. N.Y. Acad. Sci. 92, 651 (1961).

19. The absorption rate of subcutaneously injected insulin and the blood sugar concentration. CHR. BINDE]~, AA. V. NIELSEN and K. J~I~GENSEI~. Hvi- dore Hospital & Novo Research Insti tute, Copenhagen.

VoL 2, No. 2, 1966 Organization Section �9 Abstracts 14I

Four types of pharmaceutical insulin containing iesI-insulin were prepared: Insulin Novo, Actrapid, Lento and t~apitard. The insulins were given subcutaneously to diabetic patients. The absorption rate was estimated from external registration of the radioactivi ty above the site of injection. Blood sugar concentrations were determined simultaneously with the radioactivity measurements. The neutral solution, Insulin Actrapid, was absorbed faster than the acid insulin solution, Insulin Novo. Significant correlation existed between the absorption rate and the corresponding change in blood sugar concentration. In- sulin Lento was absorbed faster from the abdominal region than from the femoral region. The absorption rates for l~apitard were almost the same for the two regions. Absorption rates of Lento and Rapi tard and changes in blood sugar concentrations were significantly correlated in the periods from 8 a.m. to 11 a.m., 11 a.m. to 3 p.m., and 3 p.m. to 9 p.m.

20. Effects of insulin in vitro on adipose tissue from patients with diabetes mellitus. P. BJOtCNTOI~P. First Med. Service, Sahlgrenska Sjukhuset, G6teborg, Sweden.

A system has been worked out for human adipose tissue, where insulin in physiological concentrations in vitro causes an increase in the incorporation of label from 1-i4C- glucose into carbon dioxide, glyeeride-glycerol and fa t ty acid release. 10 controls were compared with 9 insulin- dependent diabetic patients. The adipose tissue of diabetic patients showed an increase in glycerol and free fa t ty acid release, which could be inhibited with insulin in vitro, a decreased incorporation of label into glyceride- glycerol, and a severely decreased synthesis of fa t ty acids, which could not be corrected by the addition of insulin in vitro.

21. Beziehungen zwischen Kalium- und Kohlehydrat- stoffwechselstfrungen bei Diabetes Mellitus. K. BoJAsrowicz, A. ZVBOWSKI und T. PATO~A. I. Medi- zinische Klinik der Medizinischen Akademie in Lodz, Polen.

Bei 84 Personen mit stabilem Kohlehydratstoffwechsel wurde die Glueosetoleranz and ihre Beziehung zum Ka- liumspiegel im Serum, Plasma und in Erythrocyten untersucht. Bei 43 Fettsiichtigen wurde mehrmals oral 2 mg eines Saliurcticums (Renese, Pfizer) verabreicht. Die da- nach auftretende Hypokali~mie verursachte eine Vermin- derung der Glucosetoleranz, die bei 23 Kranken mit Dia- betes mellitus am ausgepr~gtesten war. Bei 12 Kranken mit la tentem Diabetes wurde eine mittel-starke und bei 8 Fettsiichtigen ohne Kohlenhydratstoffwechselst6rungen nur eine schwache Verschlechterung der Glucosetoleranz gefunden. 36 Pat ienten mit Hypokali/imie erhielten eine Kaliumaeetat und Bromkalinmmisehung his zur Erlan- gung der Normokali/~mie. Daraufhin normalisierte sich die Glucosetoleranz. Die gleichzcitige Verabreichung yon Renese und der Kaliummisehung in I5 F~llen fiihrte zu einer unbedeutenden Erniedrigung des Kaliumspiegels ohne wesentliche Glyk/imiever/inderungen. Beim Gegen- regulationsdiabetes erreiehte man nach Anwendung der Kalinmmischung allein bei 33 von 36 Behandeltcn eine deutliche Verbesserung der Glucosctoleranz und gleichzeitig einen geringgradigen Anstieg des Kaliumspiegels im Serum. Bei 5 Gesunden waren die Ver/inderungen unwe- sentlich. Sgmtliche Ergebnisse sind statistisch gesichert. Schlugfolgerung: Bei Beurteilung des latenten Diabetes und des Diabetes mellitus sollte der m6gliche Einflul3 einer St6rung des Kalinmstoffwechsels mit in Betracht gezogen werden.

Relationships between disturbances in potassium metabolism and in carbohydrate metabolism in diabetes mellitus.

K. BOJANOWICZ, A. ZUBOWSKI and T. PATORi. First Me- dical Clinic of the Medical Academy; Lodz, Poland.

The glucose tolerance was measured in 84 controlled persons and related to potassium concentrations of serum, plasma and erythrocytes. Forty-three obese patients were given 2 mg of l~enese (Pfizer) by mouth for 7 days to promote elimination of salt. The resulting hypokaliaemia caused a reduction in glucose tolerance. This was greatest in 23 patients with diabetes mellitus, moderate in 12 patients with latent diabetes, and least in 8 obese patients with normal carbohydrate metabolism. Thirty-six hypo- kaliaemic patients were given a mixture of potassium acetate and potassium bromide for an average of days until serum potassium was normal. This brought about a return to normal of the values obtained in the glucose tolerance test. Simultaneous administration of l~enese and the potassium mixture in 15 cases caused only an insignificant fall in serum potassium without any appre- ciable change in blood glucose. In 33 out of 36 patients with counterregulatory diabetes, the administration of the potassium mixture alone caused a noticeable improvement in the glucose tolerance test and at the same time a small rise in serum potassium. In 5 normal subjects, the changes were unimportant. The results obtained were statistically significant. Conclusion: In the assessment of latent diabetic states and of diabetes me]litus, the role of disturbances in potassium metabolism should not be overlooked.

22. Insulin therapy beyond normoglycemia in the treatment of diabetes and the diagnosis of prediabetes. C. BOlVESSA, L. CREI~O:NINI and PAD. PASlNI. 0sp'edalo Policlinico, Is t i tut i 0spitalieri, Milano, Italy.

The limited success of insulin-induced restoration of normoglycemia in the t reatment or prevention of vascular or obstetrical complications of diabetes mellitus suggests the existence of an insulin deficiency which is not evidenced by hyperglycemia, and therefore not corrected by insulin t reatment aiming at the normalization of blood glucose concentrations. The authors show that insulin adminis- trat ion beyond the doses required for the restoration of normog]ycemia is the only means to compensate for this insulin deficiency and that such a t reatment is well to- lerated.

23. Adaptation of epididymal adipose tissue hexokinase to carbohydrate diet. B. BORI~EBmK. Inst i tute for Medical Biochemistry, Oslo University, Norway.

The effect of dietary carbohydrate (glucose) on epididymal adipose tissue hexokinase activity was studied. A rela- t ively low hexokinase act ivi ty was found in 48 hour fasted rats. When sueh fasted rats were refed a high carbohydrate diet ad libitum, hexokinase activity increased slowly until a maximal value was reached at about 36 hours after start of the refeeding. Kinetic experiments indicated the presence of two distinct hexokinascs with different Km(glucose)-values. Both enzymes adapted to the carbohydrate diet. The adaptation of hexokinase act ivi ty to glucose was observed in vitro, in the epididymal adipose tissue homo- genate as well as in the incubated whole fat pad. I t was found tha t insulin was a necessary factor in the process.

24. Kohlenhydrat- und Fettstoffwechsel unter oraler Ge- stagen- Oestrogen-Medikation. P. BOTTERMANN, K. KOPETZ, K. SCHLEYPEN und K. S c H w ~ z . II . Medizinische Universit/its-Klinik, Miinchen.

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142 Organization Section �9 Abstracts Diabetologia

Uber j eweils 3 Zyklen wurden insgesamt 17 Probandinnen zwei verschiedene Gestagen-Oestrogen-Kombinationen verabreicht. Vor und w~hrend dieser Medikation fiihrten wit i.v. Glucose-Belastungen durch und ermittelten die Glucosetoleranz an Hand des Glueoseassimilisationskoef- fizienten K. Gleiehzeitig verfolgten wir die Konzentra- tions/inderungen des Seruminsulinspiegels (immunolo- gisch) und der nichtveresterten Fettsauren. Wir fanden keine signifikanten Anderungen der genannten Parameter und konnten uns somit im Gegensatz zu Gershberg und Mitarbeitern yon einer diabetogenen Wirkung yon Ge- stagen-Oestrogen-Gemischen nicht iiberzeugen.

Carbohydrate and fat metabolism dm'ing oral gestagen- oestrogeu therapy. P. BOTTERMANN, K. KGI'ETZ, K. SClILEYI~EN and K. SCHWARZ. 2nd Medical Clinic, Munich.

Two different gestagen-oestrogen combinations were administered for 3 menstrual cycles each to a total of 17 women. Intravenous glucose tolerance tests were performed before and after treatment and the glucose tolerance was determined by means of the glucose assimilation coefficient K. At the same time, the changes in serum insulin concentrations were followed immuno]ogically and the free fatty acids were also determined. There was no significant change in these parameters. Therefore, contrary to Gershberg and co-workers we were not able to establish a diabetogenic effect of gestagen-oestrogen combinations.

25. Les hypoglyc6mies post-alcooHques de l'adulte: consi- d6rations cliniques, biologiques et physiopathologiques: 6rude de 8 observations. H. Bowl~, G. DOICF, M. TUTI>T et Y. ]~IQUOIS. Chaire de l~utrition humaine, H6pital de l 'H6tel-Dieu; Paris.

Confirmant 1as observations ant6rieures, l'hypoglyc@mie survient chez des alcooliques ddnutris, sous aliment@s, l 'occasion d'une absorption exagdr@e d'alcool. L'expres- sion clinique enes t trompeuse, les manifestations d'ivresse dtant prddominantes 5 fois sur 8. L'exploration biologique statique met en ~vidence l'hypoglycdmie, souvent au~ dessous de 0 g 40/1, l 'dldvation de l'aleooldmie 2 g/l, l'dl@vation du rapport lactate/pyruvate, une non r@ponse au glucagon. L'insulindmie a @t@ trouv@e normale dans les 2 cas ex- plords. La ponction biopsie hdpatique chez 4 malades a montrd la prdsence de glycog6ne mais sa r@partition anormale dans le tissu hdpatique. Line @rude dynamique du mdtabolisme de l'alcool, par perfusion de 180 g d'alc0ol en 8 heures, a dr@ mise en oeuvre avec le concours de J. Tremoli~res, dans 2 accidents hypoglyedmiques et chez 8 alcooliques chroniques normoglycdmiques, de faqon & essayer d'@tablir les m@- canismes physiopathologiques de l 'dthyloxydation. Les rdsultats sont discut~s. I1 apparalt essentiellement qu'il existe chez les alcooliques chroniques 2 voles accessoires de la d6gradation de l'alcoo]: l& voie peroxydasique et ]a voie 1%. N. asique. Ces voles ne seraient pas ddvelopp@es chez les rares malades souffrant d'hypoglye~mie. II en rdsulterait un trouble de la glyeog@nolyse et une diminution de la production hdpatique de glucose.

Post-alcoholic hypoglycaemia in the adult: clinical, biochemical and physiopathologieal considerations: study of eight cases. H. Boule, G. DORF, M. TURIN and Y. I:~IQUOIS. Dept. of Human ~utr i t ion, HStel-Dieu, Paris.

Hypoglycsemia was found to occur in malnourished and undernourished alcoholics when they consumed an excessive amount of alcohol. This confirms previous observations. The clinical appearance was deceptive, since

in 5 out of 8 eases the predominating signs were of drunkenness. A biochemical examination showed hypoglycaemia, often below 40 mg per 100 ml, a rise in the alcohol level to more than 2 g/l, a rise in the lactate/ pyruvate ratio, and a failure to respond to glucagon. In the 2 cases where the insulin level was examined, it was found to be normal. Liver puncture was performed in 4 patients and showed that glycogen was present but was distributed in an abnormal way through the hepatic tissue. A study of alcohol metabolism by perfusion of 180 g in 8 hours was carried out, with the help of J. Tremoli@res, in two victims of hypoglycaemie accidents and in eight normoglyeaemic chronic alcoholics, with a view to determining the physiopathologieal mechanisms of hypoglycaemia during alcohol absorption. The results are discussed in the paper. In essence, there appear to exist in chronic alcoholics two accessory pathways of alcohol breakdown : one employing peroxidase , the other l%NAse. These pathways are not developed in the rare patients who suffer from hypoglycaemia. The effect is to cause disturbances of glycogenolysis and a fall in the hepatic output of glucose.

26. On the application of the pyruvate kinase reaction for studies of phosphorylation. S.E. BRoy, I~, G. W E ~ E ~ M ~ K and E. Bo~G~uND. De- partment of Histology, University of Uppsala, Sweden.

Development of ultramicrochemica] methods is required for studies of the pancreatic islets in animal research and for the assay of various biopsies from human diabetics. The pyruvate kinase reaction (ADP ~ phosphoenolpyruvate = ATP ~- pyruvate) permits measurements of small amounts of ADP. With a coupled lactate dehydrogenase reaction the consumption of ADP can be followed by the conversion of NADII to NAD*. The application of this analytical system for studies of phosphorylation from ATP makes it possible to maintain the concentration of ATP at a low level due to cyclic regeneration. The applicability of the pyruvate kinase reaction has been evaluated by kinetic studies on the phosphorylation of different sugars, l~elevant inhibitory effects have been tested to settle the appropriate concentrations of the reactants and other additives. The results seem also to be of interest for om�9 conception of the regulation at the substrate level. Interference of ATP-ase and action of phosphatases on phosphoenolpyruvate constitute sources of error in analyses of crude tissue preparations; avoid- ance of these effects is thus essential.

27. Spontaneous hypoglycemia in diabetes after long-term treatment with sulphonylureas. B. BR~I. Maria Vittoria Hospital, Torino.

There is good evidence that the hypoglycemic effect of sulphonylureas is primarily referable to a stimulation of insulin secretion by the /~-cells, even though the blood sugar level is reduced. During long-term treatment of adult diabetes with sulphonylureas one therefore might advance the theoretical question whether an extremely protracted hypersecretive stimulus could eventually lead not only to an exhaustion of the islet-cell apparatus ("secondary failure"), but equally to a functional hyperinsulinism with clinical hypoglycaemie syndrome. These considerations are dictated by the personal observation of 3 cases characterized by the following clinical features, in chronological sequence: i) Well-established diagnosis of diabetes of medium severity in women (aged: 50, 65, 68); 2) Continuous treatment with tolbutamide over a period of at least 18 months and suspended a fortnight before the observation; 3) Absence of diabetic syrnptomat-

Vol. 2, No. 2, 1966 Organization Section �9 Abstracts 143

elegy in the last 6 months ; 4) Spontaneous hypoglyeemic syndrome with neurological disorders (transient hemiple- gia in 2 eases) relieved by sugar administration; 5) Serial determinations of fasting blood sugar gave values under 90 mg~ (Hagedorn-Jensen) and intravenous tolbutamide test showed the persistance of marked hypoglycemia at the second and the third hour. Owing to the uncertain results obtained with the plasma insulin assays and in the absence of any anatomical control, such cases could be ascribed, in a purely hypothetical way only, to the group of hypoglyeemie syndromes related to diabetes. Islet-cell neoplasia as a compensatory phenomenon both in man and in experimental animals is well-kown.

28. Observations on factors influencing lipolysis in vitro. THOMAS W. BUI~NS, C.N. HALES and ANNE STOCKELL I'IAICTREE. Dept. of Biochemistry, Univ. of Cambridge, England.

We have tested human sermn, various hormones and human pitui tary fractions for lipolytic act ivi ty using in- tact epididyrnal fat pads of the rat and isolated cell suspensions prepared from rat fat pads and from human adipose tissue. Glycerol release was enhanced when serum was incubated with epididymal fat pads and with rat isolated adipose tissue cells, but no difference in this effect was noted between samples obtained in the fasting state compared with those obtained �89 hour after glucose administration. Suspensions of isolated cells from human adipose tissue were responsive to adrenaline, glucagon and to human pitui tary peptide and protein fractions, including ones with high TSI-t, H G t t and ACTt t content. Site of origin of the tissue and age of the donor appear to be factors influencing the responsiveness of cell suspensions. In general, susgensions of rat cells were more responsive, on a weight basis, to these human pituitary fractions, than were suspensions of human cells.

29. Glycosuria and the diagnosis of diabetes. W . J . H . BUTTERFIELD, I{. KEEN and M.J. WHICHELOW. Department of Medicine, Guy's Hospital Medical School, London, S.E. 1.

The presence of glycosuria post-prandially or in a random urine sample has long been used as a tool in the diagnosis of diabetes, routinely and in mass screening surveys, but evidence is accumulating that this simple procedure may be inadequate. Standard 50 g glucose tolerance tests were performed on approximately 300 subjects in the age/sex stratified random sample drawn from the population during the Bedford Diabetes Survey. Urine specimens were tested for glycosuria between 60 and 90 minutes after glucose. Whereas in the survey 4% of this sample had had post prandial glycosuria, 30% of them had glycosuria during the glucose tolerance test. Analysis of the results shows that the peak and 2 hour blood sugars tend to rise with age, and that the level at which glycosuria appears also tends to rise with age. This is particularly striking amongst the women. The value of the findings of glycosuria following 50 g glucose or post-prandially in relation to the diagnosis of diabetes will be discussed.

30. Glucose disposal in the intravenous glucose tolerance test and its realtion to k-values. W . J . H . BUTTE~FIELD, M.E. ABaA~S, J .S . WA~ELIN and M.J. WmC~ELOW. Depar tment of Medicine, Guy's Hos- pital Medical School, London, S.E. 1.

To investigate the validity of K-values in the intravenous tolerance test, glucose uptake has been followed in the

forearm tissues during the test in lean and plmnp non- diabetic subjects and diabetic patients. Arterial and venous blood sugar levels and forearm blood flows were measured continuously for 8 minutes from the start of the glucose infusion, and then at frequent intervals until 1 hour. The mean glucose uptake during the hour varied widely amongst the non-diabetics (range 0.277 to 1.442 mg/ 100 ml/min) being greater in the lean subjects. A close inverse correlation was found between cell glucose uptake and fat fold thickness (as a measure of obesity) (R = --0.855, 0:01 > P > 0.001). In the diabetics glucose uptake was lower than would be expected for their degree of fatness. Calculations have also been made of the total glucose uptake in the peripheral tissues, the amount of glucose remaining in the Mood and the amount being excreted in the urine. K-values have been derived by computer from both the absolute and incremental arterial and venous blood sugar curves, and comparisons made between diabetics and non-diabetics. The relationship between K-values and the disposal of glucose into the various body compartments will be discussed.

31. Glucose tolerance test in normal and obese subjects with low plasma free fatty acid levels experimentally induced by nicotinic acid. U. BUTTUI~INI, A. GNUDI, C. COSOELLI, G. VALENTI and V. PALlV[ARI. Ist i tuto di Semeiotica Medica, Universi ty of Parma, Italy.

According to the assumptions of some authors (Randle, Vannotti) the increased levels of plasma FFA impair the glucose utilisation by muscle and adipose tissue "in rive". In previous research we have observed, on the contrary, that in obese subjects the glucose tolerance is still altered when the plasma FFA levels decrease to normal values after glucose administration. In our present work we have experimentally lowered the plasma FFA level in normal and obese subjects by carrying out a perfusion of nicotinic acid during a venous glucose tolerance test. No statistically significant differences have been observed in normal subjects, and obese subjects with abnormal G.T.T. The authors conclude that it is difficult to explain the abnormalities of the carbohydrate metabolism of the obese subject as depending upon only a primary trouble of plasma FFA.

32. Effect of dicumarol (bishydroxycoumarin) on the penetration of hexoses and pentoses into the cut diaphragm. J.L.I~. CANDELA, R. ROSALES, M. SALINAS and the tech- nical assistance of D. I'IEI~MOSO and M. REUS. Institute "G. Maranon"; Madrid.

Results obtained in other experiments prompted us to determine whether Dicumarol has any effect on the penetration of sugars into the "cut" diaphragm. Diaphragms were incubated in Krebs-phosphate buffer supplemented, according to the particular experiment, with D-glucose, D-ga]actose, D-3-O-methylglucose, D-xylose or L-arabinose. Dicumarol dissolved in 1 ~o Na0I-I was added to the buffer to give a final concentration of i • I0 -a -- 1 • i0 -a M. When used, insulin was at a concentration of i0 000 #U/ml. The final pI-I was 7.0. Dicumarol inhibited the passage of glucose and of galactos into the muscle and also inhibited the effect of insulin. This inhibition was not observed with D-xylose and L- arabinose, where it appeared that the passage of these pentoses was accelerated. The results of these and subsequent experiments will be discussed.


33. The effect of inhibited FFA-mobilization on plasma beta-hydroxybutyric acid. L.A. CARLSON and J. 0STeAls. Dept. of Internal Med., Karolinska Hosp. and King Gustav V l~esearch Inst., Stockholm, Sweden.

Among events secondary to the excessive mobilization of F F A in various states is the increased production of ketone bodies and triglycerides (in the liver), which may lead to hyperketonemia and hyperlipemia. The purpose of the present investigation was to study whether a decrease in the mobilization of F F A from adipose tissue would per se lower an elevated concentration of beta-hydroxy-butyric acid in plasma. For that reason the acute effect of nicotinic acid administration on plasma lipids, plasma beta- hydroxy-butyric acid and blood glucose was studied in fasting subjects and diabetic patients.

34. Insulin and growth hormone in plasma during glucoses infusion in turner's syndrome. E. CEICASI, J. LINDSTEN and 1%. LUFT. Department of Endocrinology, Karolinska sjukhuset, Stockholm.

Prolonged glucose infusion in 11 non-diabetic patients with Tamer ' s syndrome was accompanied by blood glucose levels higher than normal. The plasma insulin response to hyperglycemia was, in most instances, of the delayed type. Plasma growth hormone reacted abnor- mally to hyperglycemia in many of the patients. There was no increase in the frequency of diabetes among the close relatives of the patients in a large sample.

A mathematical model was constructed of blood sugar curves (obtained by means of an oral loading dose of 50 g glucose), the number of curves used as a base being 150:50 from normal subjects, 50 from known diabetics, and 50 from latent diabetics. Each type is characterized by a eonsistent curve representing a mathematical model corresponding to the appropriate type of diabetes, and each curve is expressed by 6 figures, each of which can be represented by an individual point on an axis in a n abstract six-dimensional system or by a vector drawn from the beginning of the system towards the position of the individual point. This gives a multidimensional space formed by the aggregate of the points representing the different types of blood sugar curves. An ellipsoid can be found relating this space to an equal probability density. Baye's formula enables the degree of probability to be calculated, whereby it becomes possible to consider any curve as representing one of the 3 types of curve, provided that the charac- teristic of the aggregate of points by equations of the extreme surfaces and a calculation of the functions of the probability density are available. Verification of this mathematical model by a data processing-machine in the practical diagnosis of i20 unknown eurves showed a 90~o agreement with the clinical diagnosis. The use of a mathematical model has emphasized the importance of investigating the blood sugar curves in the obese and in patients treated with chlorothiazide. I t pro- vides an objective and efficient method of detecting latent disturbances of carbohydrate metabolism and of differentiating latent from other forms of diabetes.

35. Essai d'application d'un module math6matique dans le diagnostic pr6coce du diabgte. J. CH5EBOWSKI, J. WA~TA~ et T. BOGDANIK. II . Clinique mddicale de l'Acad@mie de M@decine, Bialystok, Pologne.

Le mod@le mathTmatique des courbes glyeTmiques (obtenues apr@s surcharge de 50 g de glucose per os) est form@ ~ partir de 150 eottrbes individuelles: 50 normales, 50 obtenues chez des diab@tiques dTclarTs et 50 chez des diabTtiques latents. Chaque type de courbe est earact@ris6 par une 6quation conforme qui repr@sente un modgle mathTmatique correspondant ~ un type donn6 de diabgte. Chaeune de ees courbes est exprimTe par 6 chiffres rTels, dont chacun peut &tre reprTsent6 par un point individuel s u r u n axe dans un syst~me abstrait de 6 dimensions ou par un vecteur dress@ du commencement du systgme vers la position de ce point individuel. On obtient ainsi un espace multidimensionel form@ par une sgrie de points reprgsentant les diff@rents types des courbes glycTmiques. On peut trouver une ellipsoide ap- proximative de cet espace ~ une densit@ de probabilit6 6gale. La formule de Bayes permet de calculer le degr6 de probabilit6 avec lequel il est possible de consid@rer tme courbe quelconque eomme repr@sentative d 'un des 3 types de courbe, pour autant que nous ayons ~ disposi- tion la caractTristique de l 'ensemble des points par des 6quations des surfaces extrSmes et le calcul des fonctions de la densit6 de probabilitT. La vTrification de ce modSle mathTmatique par une machine ~ calculer dans le diagnostic pratique de 120 diff@rentes courbes inconnues, a montr6 un accord de 90~o avee le diagnostic clinique. L'usage du mod@le mathTmatique a soulign6 l ' importance de l 'exploration des courbes glyeTmiques chez les obgses et ehez les personnes recevant des ehlorothiazides pour le dTpistage objectif des troubles latents du mTtabolisme hydrocarbon@ et pour le diagnostic diff@rentiel du diabgte latent.

Attempt at applying a mathematical model to the early diagnosis of diabetes. J. CttLEBOWSKI, J . WAtCTAK and T. BOGDANIK. 2rid Med. Clin., Academy of Med., Bialystok, Poland.

36. The antilipolytic action of insulin. C. CIILOUVERAKIS. M. R. C. Metabolic Reactions Research Unit, Department of Biochemistry, Imperial College of Science and Technology, London.

Many of the metabolic effects of insulin can be at tr ibuted to its primary action on glucose metabolism. A notable exception appears to be the inhibitory effect of insulin on the epinephrine-stimulated lipolysis by rat adipose tissue. In the present paper, the results of studies on the effect of insulin on lipolysis and its modification by glucose analogues, such as 2-deoxyglueose, will be presented. Insulin was found to be an antipolytic agent even in the absence of glucose whereas 2-deoxyglucose inhibited the reesterification rate of NEFA. Further studies on the action of insulin on the extracted lipolytic anzyme of the rat adipose tissue and on its possible mechanism will be presented and discussed.

37. The relationship between immunochemical serum insulin and muscle glucose uptake in the human forearm. N. ffUEL CHRISTENSEN, H. ORSKOV and K. LUNDB2EK. The 2nd Clinic of Internal Med., Kommunehospitalet , Aarhus.

We have investigated the effect of endogenous and ex- ogenous human insulin on glucose uptake in muscle tissue. The investigation has been performed on non-diabetics and on untreated juvenile diabetics. Blood samples were collected from the brachial artery and from a deep musele vein. Serum insulin ( t t ~ . s and RANDSE) and blood glucose (glucose oxidase) were determined in the fasting state, during i .v. glucose infusion, as well as after i.v. injection of human insulin (NOVO). Blood flow was estimated by classical venous occlusion plethysmography. The following results are discussed: 1) In human forearm studies, a linear relationship between muscle glucose-uptake and immunological serum insulin was obtained i n non-diabetics and in two un-

Vol. 2, No. 2, 1966 Organization Section ' Abstracts 145

treated, juvenile diabetics. But one juvenile diabetic did not respond to insulin. 2) Whole-body insulin utilization studies after injection of non-radioactive human insulin (NOVO) showed insulin half-life values between five and fifteen minutes.

38. The effect of glucagon and secretin on glucose disposal rate and serum immunoactive insulin. M. C~RISTEN-SEN. t tv idore Hospital, N e r o Research Inst., Copenhagen, Denmark.

There is evidence that both glucagon and secretin, given intravenously with glucose, augment the insulin response and thereby the glucose disposal rate. 50 normal female subjects were subjected to the double intravenous glucose tolerance test. The effect of glueagon and secretin given with the second glucose injection has been investigated. A significant rise in the glucose disposal rate has been shown with both substances in the same person. The effect on serum immunoactive insulin, and the physiological aspects will be discussed.

39. M@tabolisme in vitro de l'ac@tate dans le tissu h@pati- que de la souris ob~se-hyperglyc@mique de bar-harbor (O-H) jeune. J. CIIRISTOI)HE, J. FUI~N-ELLE et J. WIlglkND. Laboratoire de Chimie biologique et de la Nutrition, Faeultd de Md- deeine et de Pharmaeie, Universit@ libra de Bruxelles, Belgique.

I. La concentration des phospholipides est normale mais cello des triglyc6rides d6passe 5 fois la normale dans le foie de la souris O-H. Les glyc~rides, phospholipides et F F A du foie O-H eontiennent plus de 18 : 5 (acide ol6ique) et moins de 18:2 et d'acides gras longs (20 C et plus) qua los lipides eorrespondants chez la souris N (non obese t@moin). II . Une tranehe de foie O-H, incub6e 90 rain dans un tampon de Landau, Hastings et Zottu (K+ 110 #Eq/1) contenant une albumine pauvre en F F A (40 g/l; 2 Eq/M), du glucose 10 mM et de l'ac@tate-l-14C 0.10 mM (29 e/M) est avide de 58:2 et e~de 18:1 au milieu. En outre le tissu O-t{ incorpore l 'ae6tate-l-l~C 2 ~ 3 fois mieux dans les glycgrides et aussl bien dans les phospholipides qua le tissu N. D'une mani@re g~ndrale le foie O-H synth6tise in vitro moins d'aeides gras courts et plus d'acides gras k 18 C e t d'acides gras monod6satur@s (56:5 et 18:1). L'act ivi t6 sp@cifique (a.s.) des F F A h6patiques est moins 61ev6e chez la souris O-H car le peti t pool des F F A in- tratissulaires est un peu plus ddvelopp~ que chez l'ani- real N. III. En presence d'une albumine satur~e en FFA (7 Eq/M) l'incorporation d'acdtate dans les aeides gras des glycdri- des est moins ralentie dans le foie O-I-I que dans le tissu N. IV. Aussi bien chez la souris N que ehez la souris O-H, l'incubation avee eerie albumine saturde en FFA n'affeete pas l'oxydation de l'ac@tate-5-1tC, mais elle stimule la ddsaturation des aeides 16:0 et 18:0 ndoform@s ainsi qua l'61imination des d6riv@s 16 : 1 et 18 : I dans le milieu. L'a.s. des FFA h6patiques diminue, ce qui tient k une moindre lipogen@se, h la perte des FFA ndoform@s et k un affiux de FFA non radioaetifs (18 : 2) par example).

Metabolism in vitro of acetate in hepatic tissue of young bar-harbor (0-H) obese hyperglycaemic mice. J. C]~RIS~OPEE, J . FU~N-EL~E and J . WIN-AND. Biochemis- t ry and Nutri t ion Lab., Free Univ. of Brussels, Belgium

I. In the O-H. mouse liver the concentration of phospholipids is normal but the concentration of triglycerides is five times the normal. The glycerides, phospholipids, and

non-esterified fat ty acids (NEFA) of the O-H liver contain more 18:1 (oleic ac id )and less 18:2 and long-chain fat ty acids (20 carbon atoms and more) than the corresponding lipids in the N mouse (a non-obese control). II . A section of O-H liver incubated for 90 minutes in a Landau, Hastings, and Zottu buffer solution ( K + 110 #Eq/1) containing albumin, poor in N E F A (40 g/l; 2 Eq / M), 10 mM of glucose, and 0.10 mM (29 c/M) 1-i4C-acetate showed avidi ty for 18:2 and yielded up 18:1 to the medium. Moreover, the O-H tissue incorporated 1-1*C-acetate twice to three times better in the glycerides than N tissue and just as well in the phospholipids. Generally speaking, O-It liver synthetized in vitro fewer short-chain fat ty acids and more 18-carbon fat ty acids and monodesatu- rated fat ty acids (56:1 and 18: 1). The specific activity of the hepatic non-esterified fat ty acids was lower in the O-H mouse, for the small pool of tissue N E F A was a little more developed than in the N animal. I I I . In the presence of albumin saturated in non-esterified fat ty acids (7 Eq/M) the incorporation of acetate in the fat ty acids of the glyeerides was slowed down less in O-t{ liver than in N tissue. IV. Both in the N mouse and in the O-t t mouse, incubation with the albumin saturated in non-esterified fat ty acids did not affect the oxidation of 5-1aC-acetate, but it stimulated the desaturation of the newly formed 16:0 and 18 : 0 acids as well as the elimination of the 16 : 1 and 18:1 derivatives from the medium. The specific act ivi ty of the hepatic non-esterifid fa t ty acids decreased, due to reduced lipogenesis, loss of newly formed non-esterified fat ty acids, and influx of non-radioactive N E F A (such as 18:2).

40. Conjugal diabetes. A.M. CooK~., M.G. FItzGErALD, J.M. MALIN-s and D.A. PYXE. Rare Diseases subcommittee of the Medical and Scientific Section, British Diabetic Association.

A co-operative study has been made of the children of conjugal diabetic couples. Of the children of 250 eouples, both of whom were diabetic, about 5~o have been diag- nosed as having diabetes. This is a greater number than was found in control couples, in whom neither or one parent was diabetic. Diabetes is commoner in the children of diabetic parents who develop diabetes early in life than in those of older couples. If diabetes depends upon a single recessive gene all the offspring of the conjugal diabetic pairs should eventually become diabetic. However, the incidence so far suggests that not more than one quarter will do so.

41. Plasma free fatty acids (nora) during prolonged glucose or insulin infusion in normal and diabetic subjects. C.A. C•AVETTO, L. SOIANZO, A. SikIAN-I, P. MARTIIgO and A. VITEI~LI. General Medical Clinic of the Universi ty of Turin.

25% glucose solution (0.5 g/min) was given by intravenous infusion lasting four hours to normal subjects and adult diabetics, according to the experimental technique of Ceresa etal . (Diabetologia 1, 73 (1965)). The plasma N E F A levels determined every 15' throughout the infusion by the method of Dole (J. elin. Invest. 35, 150 (1956)). In the normal subjects the plasma N E F A levels showed a sharp fall in the first and second hours, followed by only very slight further variations or, at the most, by a slight increase. In the diabetics there was a slight drop of the plasma N E F A during the earlier period of infusion, and a well-marked drop during the later period. In most instances, the lowest level of N E F A was associated with


the highest level of blood sugar. Insulin was administered intravenously at the rate of 0.05 u/rain for three hours and the plasma N E F A levels were determined every 15'. In the normal subjects the plasma N E F A level dropped considerably during the first hour, and. then rose again. The behaviour of the plasma N E F A level in the diabetic subjects was similar to that in the normal, but the drop during the first hour was less great. The behaviour of the plasma N E F A level was similar to that of the blood sugar. The differences found between the behaviour of the plasma N E F A level in normM and in diabetic subjects prob- ably depend on the different carbohydrate metabolic pat- perns in the two groups tested. The late fall of the N E F A level during the later period of glucose infusion in the diabetic subjects is still difficult to explain.

42. Serum levels of uric acid in diabetes mellitus. G. CREP~DI, F. SALA~I)I~, A. TIENr G. ENzI and P. AVOGARO. Clinica Medica Generale dell 'Universit~ di Pa- dova.

Serum levels of uric acid and fasting blood sugar have been determined in 124 patients with diabetes mellitus. The group of diabetic patients demonstrated mean values of serum uric acid not significantly different from those observed in a control group of 83 normal subjects (diabetic patients 5.20 ~= 1.31 rag%; normal subjects 5.27 =~ 1.15 mg%). Young diabetic patients showed serum levels of uric acid significantly lower than middle age and old patients. The significance is higher if the comparison is made according to sex. Patients with higher values of fasting blood sugar (above 250 rag%) showed lower values of serum uric acid (4.56 rag%) and vice versa (below 150 rag%, with 5.33 mg~o). The relationship has been studied between the serum level of uric acid and body weight. A positive correlation has been demonstrated in diabetic patients between serum uric acid and body weight (r ~- 0.4343; P < 0.001). An elevated frequency of athero- sclerotic diseases has been found in 15 obese adult diabetic patients with hyperuricemia (above 7 rag%).

43. Changes in plasma free fatty acids and glycaemia during some tests exploring glucose and lipid metabolism in obese patients. L. CUCUP~ACI-II, A. STI~ATA, U. ZULIA:NI, A. D~.TL'A~NA and P. CUCVRACHL Ist i tuto di Clinica Medica Generale e Terapia Medica de]l'Universit~ deg]i Studi di Parma; Italia.

The interrelationship between plasma free fatty acids and blood glucose levels in the course of tests exploring carbohydrate metabolism (oral glucose load, intravenous tolbutamide and insulin tests)was investigated in female obese patients. Decreased carbohydrate tolerance, lowered insulin sensitivity and normal responsiveness to tolbutamide, when compared with the normal control subjects, were observed in the obese patients. Moreover, an inverse correlation between the changes in plasma free fatty acids and in blood glucose after each test was found in the obese. According to these results, it seems possible to conclude that in essential obesity there exists an impairment in glucose metabolism, rather than early lipid metabolic abnormalities. Moreover, there is good evidence for the belief tha t glucose metabolism is impaired in the obese, especially considering that the lipid mobilizing effect of fasting and of epinephrine, and the postheparin clearing fsctor of plasma are normal in these patients.

44. The effects of phenformin treatment in obese patients. L. CUCURACHI, A. STRATA, U. ZULIANI and A. DELL'A~NA. Ist. di Clin. Med. Gen. e Terapie reed., Univ. degli Studi di Parma, Italy.

Phenformin caused reduction in body weight with anore- xic effect in female obese patients. In the obese with impaired carbohydrate metabolism, phenformin caused improvement in the abnormalities of some humoral in- dices by the following changes: decrease in plasma free fat ty acids and blood glucose levels; increase in glucose utilization rate (K-value) and recovery of carbohydrate tolerance and insulin sensitivity. The subjects with normal blood glucose levels and carbohydrate tolerance at the start, showed during phenformin t reatment no evident variations in glycaemia, but a decrease in plasma free fat ty acids was demonstrable.

45. Observations on the action of insulin in obese patients. L. CU~URAC~I, A. STRATA, U. ZULIANI and S. CASTELLO. Ist. di Clin. Med. Gem, e Terapia reed., Univ. deg]i Studi di Parma, Italy.

Sensitivity to insulin (intravenous injection of 0.I U/kg), evaluated by changes in plasma free fatty acids and blood glucose, was studied in female obese patients. The mean fall of the blood glucose levels following insulin injection in ~he obese was lower than that of the normal control group, while the decrease of the plasma free fatty acids was not impaired. In the obese patients the insulin effect on plasma free fatty acids was increased to an extent comparable with the reduction of the insulin effect on the blood sugar. Finally, a significant positive correlation was found between insulin responsiveness, carbohydrate tolerance (oral glucose load) and glucose utilization rate (K-value) in the obese. On the basis of these results the role of some factors inhibiting insulin activity in essential obesity is considered.

46. Leucine-induced hypoaminoacidaemia. A. CzYzYK. I I I Department of Medicine and Diabetes, Praski Hospital, Warsaw, Poland.

Normal dogs were rapidly injected with glucose, then glucose and 1-1eucine, before and after t reatment with ehlorpropamide. The blood sugar level was determined, as well as that of I]%I and of free amino acids, the last by means of column chromatography. The assimilation coefficient "Kam" was calculated for each amino acid from the curve obtained on a semilogarithmic scale. The highest coefficients after the injection of leucine were found for isoleueine, valine, tyrosine, phenylalanine, me- thionine, threonine. When ehlorpropamide was administered, the coefficients increased by 10% on the average. Only 1% of the injected leucine was excreted in the urine. The renal clearance of the other amino acids was somewhat increased, but the urinary loss remained at a low absolute value. These results point to the active transfer of many amino acids after rapid loading with 1-1eucine in the dog.

47. Influence des biguanides sur la lactacid@mie du dia- b~tique au cours de l'6preuve de charge au fructose. G. DEBRY, d-. LAUIaENT, H. VINIAKER. Groupe de Re- cherches de Nutri t ion et de Didt6tique de I ' INSERM, In- stitut ~6gional d'Hygigne, Facult6 de M@decine, Nancy.

Les auteurs @tudient l'influence de deux biguanides hypoglyc@miants, la Metformine et la Phenformine sur la lactaciddmie du diab6tique. Pour ce faire, ils utilisent ]a technique d'hyperlactaciddmie provoqude par pe r fus ion intraveineuse de fructose et la m6thode de dosage en- zymatique (lactico-deshydrog@nase) de la lactacid@mie. Cette 6preuve dynamique est pratiqu6e deux fois pour chaque malade aprgs que l'gquilibre du diabgte air @t6 r6alis6 par Fun puis par l'autre biguanide.

Vol. 2, No. 2, 1966 O r g a n i z a t i o n Sect ion . A b s t r a c t s 147

L ' d t u d e des rdsu l t a t s o b t e n u s condu i t ~ la d iscuss ion du m o d e d ' a e t i o n de ces s u b s t a n c e s hypog lyedmian t e s .

Influence of biguanides on the blood lactic acid level of diabetics during the fructose loading test. G. DEBRu J . LAU~E~T a n d H. VINIAKEm N u t r i t i o n a n d Die te t i c s 1%eseareh Group of INSERS{ , ~ e g i o n a l I n s t . of I-Iygiene, N a n c y (France) .

The a u t h o r s s t ud i ed t h e inf luence of two b iguan ides , Met- f o r m i n a n d P h e n f o r m i n , w h i c h lower t h e b lood sugar level, on t h e b lood lac t ic acid level in d iabet ics . To ca r ry ou t t h e s tudy , t h e y emp loyed t h e t e c h n i q u e of ra i s ing t h e b lood lac t ic acid level b y i n t r a v e n o u s in fus ion of f ruc tose a n d m e a s u r e d t h e b lood lact ic ac id level b y t h e lac t ic d e h y d r o g e n a s e m e t h o d . Th i s t e s t was car r ied ou t twiee for e ach p a t i e n t , w h e n his d iabe tes h a d b e e n con- t ro l l ed b y first one a n d t h e n t h e o the r b iguan ide . The s t u d y of t h e resu l t s o b t a i n e d leads to a d iscuss ion of t h e m o d e of ac t ion of t he se subs tances , wh ich lower t h e b lood sugar level.

48. Serum insulin in normal and obese persons. TOlCSTEN DECKERT a n d LEIF HP~GERU1 ~. Cogenhagen C o u n t y Hosp . , Glos t rup .

F a s t i n g s e r u m insu l in a n d glucose c o n c e n t r a t i o n were d e t e r m i n e d in 68 h e a l t h y sub jec t s , all 50 years old. 28 of these pe r sons h a d a b o d y we igh t more t h a n 15~ over t h e idea l weight . All persons h a d a b loodsuga r c o n c e n t r a t i o n be low 120 mg~ ( H a g e d o r n N o r m a n Jensen) . F o r insu l in d e t e r m i n a t i o n t h e i m m u n o l o g i c m e t h o d of Ha le s a n d R a n d l e was used. T h e r e was no dif ference in insu l in con- c e n t r a t i o n b e t w e e n m e n a n d women . 2 5 % of t h e obese pe r sons h a d a s ign i f ican t ly e l eva t ed insu l in concen t r a t i on . The m e a n b lood suga r c o n c e n t r a t i o n of t h e g roup w i t h e l eva t ed insu l in c o n c e n t r a t i o n was s igni f icant ly h i g h e r t h a n t h e m e a n b lood sugar c o n c e n t r a t i o n of t h e g roup w i t h n o r m a l insu l in c o n c e n t r a t i o n . The qnes t i on w h e t h e r pe r son w i t h e l eva t ed fa s t ing insu l in c o n c e n t r a t i o n are p red i abe t i e s is discussed.

49. Der Verlauf der proliferativen Retinopathie bei nicht- hypophysectomierten Patienten. TOI~STEN DECKERT und SVE~D E. SI~O~SEN. Niels Steen- sens, Hosp i t a l , Gen tof te , D g n e m a r k .

Bei 51 D i a b e t i k e r n m i t e iner p ro l i f e r a t i ven R e t i n o p a t h i e , welche n a c h den i m A u g e n b l i e k g e l t e n d e n I~r i te r ien fiir eine H y p o p h y s e e t o m i e wohl gee ignet w/iren, wurde eine r e t r o s p e c t i v e U n t e r s u e h u n g anges te l l t . Alle F/ille w a r e n m i n d e s t e n s 2 J a h r e l ang ver fo lg t worden , im D u r e h s e h n i t t 6 J a h r e lang. 4 - - 5 J a h r e n a e h de r e r s t m a l i gen Diagnose de r pro l i fe ra t i - y e n R e t i n o p a t h i e w a r e n 5 0 % der F a t i e n t e n e n t w e d e r ge- s t o r b e n oder au f b e i d e n A u g e n b l ind . Die P rognose des S e h v e r m 6 g e n s w a r abh / ing ig 1. y o n der L o k a l i s a t i o n u n d V e r b r e i t u n g der P ro l i f e ra t ionen , 2. v o m Al te r des P a t i e n t e n u n d 3. d a v o n , ob sieh eine p e r m a n e n t e P r o t e i n u r i e en twieke l t e oder n ich t .

Prognosis of proliferative retinopathy in patients not subjected to hypophysectomy. To~s~nN DECKERT a n d SVEND ERIK SIMONSEX. Niels S teensens HOsp. Gentof te , D e n m a r k .

A r e t ro spec t i ve s t u d y was m a d e of 51 d i abe t i c p a t i e n t s w i t h p ro l i f e ra t ive r e t i n o p a t h y w ho a p p e a r e d well su i t ed for h y p o p h y s e c t o m y , as j u d g e d b y p r e s e n t l y a c c e p t e d cr i ter ia . All t h e p a t i e n t s h a d b e e n fol lowed for a t l eas t 2 years , t h e ave r ag e d u r a t i o n of o b s e r v a t i o n be ing 6 years . B e t w e e n 4 a n d 5 yea r s a f t e r t h e p resence of p ro l i f e ra t ive r e t i n o p a t h y was f irst d iagnosed , 5 0 % of t h e p a t i e n t s were

e i the r dead or b l i n d in b o t h eyes. The r a t e a t w h i c h loss of v is ion occur red d e p e n d e d on (1) t h e loca l iza t ion a n d sp read of t h e pro l i fe ra t ion , (2) t h e age of t h e p a t i e n t , a n d (3) w h e t h e r or no t p e r m a n e n t p r o t e i n u r i a developed.

50. Taux plasmatiques de l'insuline et de l'hormone de croissance (H.G.tI.) dans deux cas de diabbte lipoatro- phique. M. DEROT, G. ]~OSSELIN, I~. ASSAS et G. TCHOBICOUTSKY. H6tel-Dieu, Paris, France.

Dana la premiere observation (CAs Bow, G. BOUDIN, J.L. DE GENNES et Coll. -- Bull. Mdm. Soc. Med. llLp. Paris 114, 895 (1963)) l'insulindmie dos@e pendant une pdriode de lactescence du sdrum est de 300 /~u/ml ~ jeun. Apr~s restriction des ]ipides alimentaires en pr@sence d'un sdrum clair une hyperglyc6mie lorovoqude a dtd pratiqude par voie veineuse rapide. Le coefficient 102 K 6tait de i.i0. La glyc6mie .k jeun de 102 rag%. L'insulin6mie de 60 /~u/ml. Pendant l'6preuve l'insulindmie atteint 140 /~u/ml k la trenti6me et ~ la q u a r a n t i 6 m e m i n u t e . Le t a u x de I 'H .G .H. es t de 8 m # g / m l au d 6 b u t de l ' @ r e u v e , t a u x n o r m a l dans n o t r e l abo ra to i r e ehez u n e f e m m e a m b u l a t o i r e (Dosage r ad io - immunolog i - que avee d e u x an t i eo rps -- S t a n d a r d s W i l h e l m i N . I . H . Hs 545 A). Dans la seconde observation (Cas LEB 9 ans I/2 -- Dr. GuIAI~D, non publid ~ ce jour), l'insuline et la I-I.G.H.. plasmatiques ont dtd dosdes 30, 60, 90 et 120 minutes aprgs surcharge en glucose par vole bueeale pendant une pgriode de diab~te franc. L'insulindmie s'dl@ve progres- sivement de 90 ~ 320 #u/ml en prdsenee d'une glyc@mie croissant de 186 k 376 rag%. Le taux de l'hormone de eroissance reste dlevd pendant l'dpreuve: 25, 12 15, 25 m#g/ml, mais des taux dlevds de H.Cx.H. sont souvent eonstat@s dans notre laboratoire chez des diabdtiques sans lipoatrophie. L'hyperinsulinisme et l'hyperglycdmie semblent seeon- daires k l'hyperlipdmie, comme le suggbre l'observation Bow, oh la clarification du plasma s'aecompagne d'une dinlinution de l'anomalie de la toldrance au glucose et de l'hyperinsulinisme.

Plasma levels of insulin and human growth hormone (HGH) in 2 eases of lipoatrophic diabetes. M. DI~I~OT, G. ROSSELIN, 1%. ASSAN and G. TC~IOBI~OU~SKY. II6tel-Dieu, Paris.

In the first case (G. BOUDIN, J.L. DEGENNES el) al. Bull. Mdm. Soc. Med. m6p. Paris 114, 895 (1963))the fasting blood insulin level measured during a period when the serum was milky was 300 #u/ml. After restriction of dietary lipids and with the serum clear, hyperglycaemia was induced quickly by intravenous injection. The coefficient 102 K was I.I0. The fasting blood sugar was 102 rag% and the blood insulin 60 #u/ml. During the test the blood insulin level reached 140 #u/ml at the 30th and 40th minute. The HGH level was 8 #u/ml at the beginning of the test -- the normal level in an ambulant female subject in our laboratory (radioimmunological assay with two antibodies -- Wilhelmi N.I.II. Standards H S 545 A). I n t h e second o b s e r v a t i o n (LEB, 91/2 years old, Dr . Guia rd , as ye t u n p u b l i s h e d ) t h e insu l in a n d H G H in t h e p l a s m a were d e t e r m i n e d 30, 60, 90 a n d 120 m i n u t e s a f t e r oral glucose load ing d u r i n g a pe r iod of f r a n k d iabe tes . The b lood insu l in level rose p rogress ive ly f rom 90 to 320 #u /ml , whi le t h e b lood glucose level rose f rom 186 to 376 mg~ The H G H level r e m a i n e d h i g h d u r i n g t h e t e s t : 25, 12, 15, 25 m#g /ml . H i g h levels of H G t t , however , were o f t en obse rved in our l a b o r a t o r y in d iabe t i c s who were n o t l ipoa t roph ic . The h i g h insu l in level a n d hype rg ly - eaemia seemed to be s e c o n d a r y to t h e hype r l i paemia , as

148 Organization Section �9 Abstracts

the Boy observation suggests. In this case the clearing of the plasma was accompanied by a decrease in the ab- normality of glucose tolerance and high insulin level.

51. Int6r~t pratique et indications du dosage des acides gras libres (AGL) au cours du test au tolbutamide intraveineux. 1%. DEUIn, G. DEnOTZX, GUINA~D et C. LAUI~EI~T. H6pital Saint-Joseph (Coll~ge de M5decine des H6pitaux de Paris). Paris. Line centaine de sujets normaux et diab@tiques de toutes cat@gories ont subi une hyperglycemic provoquge par voie orale, un test k l'insuline intraveineuse et un test au tolbutamide intraveineux avec ~tude parall~le des variations des AGL. Le test au tolbutamide permet de distinguer sans ambi- guit@ quatre categories de sujets- normal, insulino-d~pendant, diab@tique obese, expos@ au risque h~r~ditaire. Los auteurs essaient de ddgager des enseignements com- pl6mentaires ~ partir des variations des AGL observ~es au cours de ce dernier test en particulier e n c e qui con- cerne: -- le d6pistage des sujets expos6s au risque h~r6ditaire, -- le bJen fond@ de certains traitements par los hypoglyc~miants oraux chez los sujets jeunes, -- la confirmation des troubles mineurs de la glycor~gu- lation observes chez certains malades non diab@tiques et difficilement interpr@tables.

Indications for and practical value of determining the free fatty acids during intravenous tolbutamide testing. 1%. DE~n , G. DE~O~X, G c I ~ D and C. L ~ E ~ T . St. Joseph Hosp., (College of Med. of the Paris Hosp.) Paris.

Approximately 100 normal subjects and diabetics of all types were given oral glucose to cause hyperglycaemia and tested with intravenous insulin and intravenous tolbutamide. The variations in the level of free fatty acids were studied concurrently. The tolbutamide test made it possible to distinguish unambiguously 4 classes of subjects: normal subjects; insulin-dependent subjects ; obese diabetics ; and subjects exposed to hereditary risks. The authors have a t tempted to draw further conclusions from the variations in the level of the free fa t ty acids, especially during the tolbutamide test. The conclusions are related to: a) the detection of subjects exposed to hereditary risk; b) the justification for certain forms of t reatment with oral compounds lowering the blood sugar level in young subjects; and c) the confirmation that there are minor disturbances of sugar regulation that are observed in certain non-diabetic patients and are difficult to interpret.

52. Insulin immunoglobulin studies in dermal hypersen- sitivity. JA~ES D~VLI~, Department of Endocrine Immunology, St. Vincent's Hospital, St. Stephen's Green, Dublin 2.

In a beef insulin treated diabetic population there is an increased incidence of dermal reactions to beef insulin. This is paralleled by a preferential binding of beef insulin by the patients serum. I t is postulated that dermal sensitivity is due to an antibody. Radioimmunoeleetrophoretic and chromatographic data will be presented, demonstrating a Ylm antibody to insulin in patients with acute local insulin reactions, and a possible relative increase in Y1 antibody in patients with dermal sensitivity.

53. Der intravenSse Glucosetoleranztest und das Verhalten der nicht veresterten FettsKuren zur Erfassung eines latenten Diabetes Mellitus.

Diabetologia

P. DIETERLE, K.P. EYIER, G.G. HOFMANI~, P.C. SCRIBA und K. SCHWA~Z. II . Medizinische Klinik der Universit~t Mfinchen, Mfinchen, DBR.

Nachdem diabetische Organver/~nderungen bereits Jahre vor Manifestation eines Diabetes mellitus auftreten k6n- non, ist die friihzeitige Erfassung eines latenten Diabetes yon groBer Bedeutung..Uns hat sich in der Diagnostik des latenten Diabetes der intraven6se Glucosetoleranztest ( i .v .G.T.T.) bewghrt. Beim i .v .G.T .T , n i t der Bestimmung des Glucoseassimi- lationskoeffizienten ka sind jedoch kG-Werte zwisehen 1.0 trod 1.2 bei uns einem unklaren Zwischenbereich zuzu- ordnen, in d e n nicht entschieden werden kann, ob die untersuchten Personen zur Gruppe der Normalen odor der ]atenten Diabetiker gehSren. Bei einer n i t d e n i .v. G.T.T. koordinierten Bestimmung der nieht veresterten Fetts~uren (NFS) l~Bt sich aus deren Verhalten aber haufig die Frage entscheiden, ob bei jenen F~llen nit kG-Werten in diesem unklaren Zwisehenbereich tats~ch- lich ein latenter Diabetes vorliegt, da bei Normalen die NFS sp/itestens naeh der 64. Minute wieder ansteigen, wahcend sic bei Diabetikern kontinuierlich bis zur 84. Mi- nute abfallen. Unsere Erfahrungen fiber die Glucoseto- leranz bei verschiedenen Erkrankungen (Adipositas, Ar- teriosklerose, Akromegalie, Hyperthyreose und Leber- krankheiten) werden kurz mitgeteilt.

The Intravenous glucose tolerance test and the behaviour of the free fatty acids in the detection of latent diabetes mellitus. P. DIETERLE, K.P. EYI~EE, G.G. HOFMAIqN, P.C. SOl%IDA and K. SCHwA~z. 2hd Med. Clin., Univ. of Munich, Ger- many.

Since diabetic changes in the organs may occur years before clinical evidence of diabetes me]litus, early detection of latent diabetes is of great importance. We have found the intravenous glucose tolerance test (i. v. G.T.T.) a useful diagnostic tool. When determining the glucose assimilation coefficient (KG) it was found, however, that KG values between 1.0 and 1.2. lay in an intermediate range of uncertain significance, and it was impossible to decide whether patients in this group were normal or latent diabetics. If determinations of the free fat ty acids (FFA) are performed at the same time as the i .v .G.T.T. , it is often possible to decide from the changes in the F F A levels whether patients with KG values in this intermediate range are, in fact, latent diabetics, since in normal subjects the F F A levels rise again by the 64th minute at the latest, whereas in diabetics they fall continuously until the 84th minute. A brief account is given of our experience with the glucose tolerance test in patients with various diseases (obesity, arteriosclerosis, acromegaly, hyper- thyroidism and liver diseases).

54. ~-Glucuronidase-Aktivit~it in Serum, Leber- und Nie- rengewebe beim Alloxan-Diabetes. 1%. E. D O H ~ A ~ u. M. K0~T.ER. Medizinische Universi- t~ts-Poliklinik, Bonn.

In Anlehnung an das auffallige Verhalten der fl-Glucuro- nidase beim Diabetes mellitus, die im Zustand der din- betischen Stoffwechsel-Decompensation cine hohe Serum- fermentaktivit~t zeigt, wird im folgenden fiber Ver~nderungen dieses Fermentes beim experimentellen Alloxan- Diabetes der Ra t te beriehtet. Es warden bier die fl-Glu- curonidase-Aktivit~t des Serums sowie des Leber- und Nierengewebes untersucht. Im einzelnen ergaben sich folgende Ergebnisse : 1. Die Serum-fl-Glucuronidase ist im Zustand der diabetischen Stoffwechseldekompensatioa stark erhSht, wobei maximal eine nahezu zehnfach hShere Akt ivi ta t als bei der Kontrollgruppe (ca. 530 7/100 ml/St) gemessen wurde.

Vol. 2, No. 2, 1966 Organization Section �9 Abstracts i49

2. Besonders hohe Serum-fi-Glucuronidase-Aktivit~iten wiesen die diabetisehen Tiere mit hochgradiger Polyurie auf. Beziehm~gen zwisehen der Serum-Aktivi tat und der HShe der Blutzuckerkonzentration resp. Harnzuckeraus- scheidung liegen sich aus den vorliegenden Ergebnissen nicht erkennen. 3. Bei der Halfte der diabetischen Tiere waren die fi-Glu- curonidase-Aktivit~iten des Leber- und Nierengewebes erniedrigv, wahrend die Aktivit~iten bei den fibrigen Ver- suchstieren den Durchschnittswerten der I(ontrollgruppe entsprechen.

~-glucuronidase activity in the serum and in liver and kidney tissues in alloxan diabetes. R.E . DOHR~A~N and M. KSHLER. Medical Policlinic, Uni- versity of Bonn.

The striking behaviour of/~-glucuronidase in patients with diabetes mellitus, indicating a high serum enzyme act ivi ty during metabolic decompensation, led us to study the changes that take place in the act ivi ty of this enzyme in rats with alloxan diabetes. Determinations of fl-g]u- curonidase act ivi ty in the serum and in liver and kidney tissue gave the following results: 1. During metabolic decompensation, the serum fi-glucuronidase levels are greatly increased, the maximum act ivi ty measured being nearly ten times that in a control gToup (about 530/~g/100 ml/hour). 2. Particularly high serum ~-glucuronidase activities were found in diabetic animals with severe polyuria. The results so far obtained do not provide any indication of a relationship between the serum activi ty and blood sugar levels or the concentration of sugar in the urine. 3. In half the diabetic animals~ the fl-glucuronidase activities of the liver and kidney tissues were diminished, while in the remaining animals the activities corresponded to the mean values for the control group.

55. Thyroiditis in juvenile diabetes. G. ENGLESON and S. A~o~ssoN. Dept. of Pediatrics, Universi ty Hosp. of Lund, Sweden.

During the last few years we have observed 4 cases of different types of thyroiditis in puberal girls with juvenile diabetes. In all cases there was an increased lability in the diabetic state, i.e. increased insulin requirement, a tendency to ketosis and unstable blood sugar values. Two girls had unequivocal Hashimoto thyroiditis, in one case there was a simple thyroiditis and the last patient had a simple goiter with thyroid antibodies and a low PBI. The 2 patients with Hashimoto thyroiditis were treated with prednisolone, whereupon the histo-pathological picture showed a tendency to normalization of the thyroiditis. There was, however, no significant clinical improvement of the diabetic condition. Data concerning thyroid function tests as well as immunological investigations, will be presented and discussed.

56. D-cells as source of a third pancreatic hormone and precursors of b-cells. A. E~PL]~. Depar tment of Zoology, Universi ty of Wash- ington.

There is now good evidence for the elaboration of a third pancreatic hormone by the D-cell which was originally described by Bloom (1931). This cell has long been con- fused with the true A-cell which produces g]ucagon, and all work in which there was no differentiation between A- and D-cells should be reconsidered. D-cells were found in all species and all animals above cyelostomes; also under various physiological and pathological conditions (e. g. hibernation, seasonal islet hypertrophy or atrophy,

hypophysectomy, starvation) D-cells are always present. There are now two hypotheses about the D-cell hormone : one suggests that it is identical with gastrin, and the other that it is lipocaie. Though there can be little doubt about the functional autonomy of the D-cell there exist in lower vertebrates islet cells which contain both D- and B-granules. Ferner (1952) postulated for human embryos the transformation of argyrophils (i.e. D-cells) into B- cells, and apparently this also occurs in seasonally hyper- plastic islets of adult toads. The question arises why transformations do not occur in adult human diabetics where D-cells are abundant, and how far lower vertebrates can be used for studies in this field.

57. A study of micturition in diabetic males and controls with ages between 20 and 50: clinical findings, cystometry, micturition-cystography and pelvic-bottom-clectromyo- graphy. S.-E. FAGERBEttG, N. KOCl~, I. PETERS]~N Ct I. STEER. Sahlgrenska sjukhuset, Universi ty of GSteborg, GSteborg, Sweden.

Urinary infections are frequent in diabetics. The cause is not clear. Neurogenic disturbances of the urinary bladder might be an important factor. Twenty-eight diabetic males have been investigated, none above 50 years of age. These patients have been compared with 25 controls. In particular, the presence of symptoms and signs from the urinary tract, and the appearance of neuropathy and diabetic angiopathy have been studied. The results of the cystometry and micturit ion-cystography showed among other things that the diabetics, even symptomless, had a large hypotone and atonic urinary bladder and sphincter disturbances in a high frequency. The pelvic bot tom electromyography showed tha t the potential durations in diabetics were significantly larger than in controls both in urethra and in anal sphincter as well as in levator ani.

58. Insulin Antibodies in patients with insulin allergy. S. FANKEAUSER and A. DE WEEK. Medical Policlinic and Allergy Research Unit, Depar tment of Dermatology, Universi ty of Berne.

Insulin allergy in diabetic patients is generally considered to be due to impurities in insulin preparations and only exceptionally to insulin itself. Insulin antibodies were determined in 16 patients with insulin allergy according to the method of Skom and Talmage. The use of corresponding labelled insulin preparations permit ted a se- parate determination of antibodies against porcine and bovine insulin. In 13 subjects a very high and in 3 subjects a moderately elevated concentration of anti- insulin antibodies was found, compared to non-allergic, insulin treated patients. Two diabetics, who were allergic to bovine insulin only, showed a higher serum binding capacity for bovine than for porcine insulin. In 5 cases insulin resistance was also observed in the course of t reatment with insulin. The frequent occurence of a high concentration of cir- culating anti-insulin antibodies in patients with insulin allergy suggests, tha t in these cases allergy is due to insulin itself and not to impurities. Insulin allergy and resistance are often observed in the same subjects, both manifestations represent abnormal immunological re- sponses to insulin.

59. Zellgcbundene Antik6rper gegen Insulin. K. FEDERLIlV, I. GIGLI, I-I. DITSCtIU:~EIT, G. I-IEINEMAI~N und E .F . PFEIFFER. Endokrinologische Abteilung der I. medizinischen Klinik und Hantklinik der J .W. Goethe Universitiit und der Stadt Frankfurt am Main.


Bei Patienten mit verz6gerter Allergic gegen Insulin wurden die wei2en Blutzellen auf ihre F/ihigkeit zur Antigen- bindung untersueht. Als Methoden dienten die Immun- fluoreszenz und die Immunadhaerenz. Bei 10 Patienten konnte gezeigt werden, dal~ Lymphoeyten und Granulo- cyten das Antigen (Insulin) zu binden vermSgen. Mit Immunfluoreszenz und Immunadhaerenz wurden nahezu gleiehe Ergebnisse erzielt. Maximal konnte eine Antigen- bindung an 60% der Zellen beobaehtet werden. Die Re- aktionsf/ihigkeit der Zellen geht der St/irke der Haut- veriinderungen parallel. Aus den Versuehen mit der Im- munadhaerenz geht hervor, dab bei der Antigenbindung an die Zelle auch Komplement fixiert wird. Neben den cytologischen Untersuchungen wurden die humoralen An- tikSrper der betreffenden Patienten best immt (Haemag- glutination und Insulinbindungskapazit/it nach BERsoN und YAnow). Teilweise wurden zellstgndige AntikSrper vor dem Auftreten humoraler AntikSrper beobachtet. ttistologische Untersuchungen der Haut am Ort der In- sulininjektion ergaben cine hyperergisehe granulomatSse Entziindung vom Typ der Sp/itreaktion. Die Untersu- chungen sind noch im Gange.

Cell-bound antibodies against insulin. K. FEDERLIN, I. GIGLI, H. DITSCItUNEIT, G. H:EINElVIANN and E .F . PFEIFFE~. Endocrinological Department of the First Medical Clinic and Dermatological Clinic of the J .W. Goethe Universi ty and of the City of Frankfurt am Main.

Using the methods of immune fluorescence and immune adherence, the antigenbinding capacity of the white blood cells was studied in patients with delayed allergy to insulin. In l0 patients, it could be shown that the lymphocy- tes and granulocytes were capable of binding the antigen (insulin). Immune fluorescence and immune adherence gave almost identical results. In some cases, up to 60% of the cells contained bound antigen. The reactivi ty of the cells paralleled the skin changes. From the immune adherence experiments it was clear that the binding of antigen by the cells is accompanied by complement fixation. In addition to the cytological studies, the ant- bodies in the blood were also determined (haemaggluti- nation and insulin-binding capacity by the method of BE~soN and YALOW). In some cases, cell-bound antibodies were observed before the appearance of antibodies in the blood. Histological studies of the skin at the site of the insulin injections showed a hyperergic granuloma- tous inflammation of the late reaction type. The studies are being continued.

60. Diminution de la sensibilit6 pancr6atique au glucose dans certaines formes de diabbte. J . P . F~LBEI~, L. G~ASSI, A.J . MOODY et A. VA~NOTTI. Clinique Mgdieale Universitaire, Lausanne.

L'dtude des relations entre les taux plasmatiques de glucose et d'insuline immuno-r@active d4montre une diminution de la r@onse insulinique aprgs surcharge glueos4e dans certaines formes de diab@te. Chez 12 cas 6tudi4s, les variations de l'insuline suivent celles de la glycdmie, mais song fortement diminu6es en intensit6, Cette ((r@sistance pancrdatique au glucose~) a dr@ reproduite expdrimentale- ment chez des sujets normaux trait4s par des injections r@pdtdes de glucagon i.m. et ehez des rats trait@s par du glucagon sous-cutan6 ou par de l 'huile d'olive per os. Lors des tests de tol4rance au glucose on observe que: 1. l'616vation m@me prolong@e de la glyc4mie produit une @14vation correspondante bien qu'attdnu6e, de l'insulin6- mie, et 2. une injection de tolbntamide en fin de surcharge glu- cos@e produit une 414ration de l'insulin4mie au-dessus des taux observ4s apr~s surcharge de glucose.

Ces observations sugg~rent la possibilit6 d'une diminution de la sensibilit4 pancrdatique au glucose plutSt qu 'un @uisement des riots de Langerhans.

Decrease in pancreatic sensitivity to glucose in certain forms of diabetes, J .P . FEr.BER, L. GRASSI, A.J . MOODY and A. VANNOTTI. Univ. 1Vied. Clin., Lausanne.

Study of the relationships between the plasma levels of glucose and immunoreactive insulin showed in certain forms of diabetes a decrease in the insulin reponse after a loading-dose of glucose. In the 12 cases studied, the variations in the insulin followed those of the blood sugar level, hut were of much lesser intensity. This "pancreatic resistance to glucose" was reproduced experimentally in normal subjects treated with repeated intramuscular injections of glucagon and in rats treated with subcutaneous glucagon or with oral olive off. During the glucose tolerance tests it was observed that : 1. even when the blood sugar level was raised for a prolonged period it produced a corresponding, though attenuated, elevation of the blood insulin level; and 2. an injection of tolbutamide after the glucose loading dose produced a rise in the blood insulin level above that observed after the glucose load. These observations suggest a possible decrease in pan- ereatic sensitivity to glucose rather than exhaustion of the islets of Langerhans.

61. Birmingham diabetes survey. A five year follow-up. M.G. FITZGERALD and L.A. P I ~ . On behalf of the Col- lege of General Practitioners of Great Britain and Ire- land.

During 1960, 18,532 subjects whose age and sex distribution was representative of the general population of England and Wales were tested for glycosuria one hour after the largest meal of the day, using glucose-oxidase test paper. Excluding 119 known cases of diabetes, 493 reported glycosuria and 465 of these had a standard 50 gram oral glucose tolerance test. In 127 an undoubted diabetic type of tolerance curve was shown. In 55 the fasting blood glucose exceeded 130 mg/100 ml. and many of these were complaining of diabetic symptoms; 72 had G.T.T. diabetes and a further 76 had an abnormal curve of doubtful significance. In addition there were 100 patients wigh lag storage curves, 54 with renal glycosuria and 108 with a normal tolerance test and no glycosuria. Five years later 49 of the 465 in the original survey had died but it has been possible to re-test some 350 in all the various groups. During this t ime about 30 new clinical cases of diabetes have appeared in the original population who had not reported glycosuria in 1960. The results of the further glucose tolerance tests will be reported and the significance of the findings discussed.

62. Pharmacological control of insulin secretion in hyperinsulinism. I-I. FRERICHS and W. CREUTZFELDT. Medizinische Uni- versitatsklinik G6ttingen.

We report on clinical and experimental observations with the nondiuretic benzothiadiazine derivative Diazoxide (3-methyl-7-chloro- 1.2.4-benzothiadiazine - 1.1 -dioxide). Three patients, each suffering from hyperinsulinism of several years duration were treated with Diazoxide. Con- servative therapy has been successful in all of the three cases, judged by the alleviation of hypoglycemie symptoms. Normoglycemia or even hyperglycemia was induced in two patients. The poor condition of all patients improved within a few weeks, so that surgery was possible.

Vol. 2, No. 2, 1966 O r g a n i z a t i o n Sect ion �9 A b s t r a c t s 151

I n eve ry case h y p e r i n s u l i n i s m h a d b e e n caused b y func- t i ona I B-cel l a d e n o m a s . L a p a r o t o m y was p e r f o r m e d a n d t he a d e n o m a s were r e m o v e d . In two p a t i e n t s t h e t u m o r s h a d b e e n local ized p r e o p e r a t i v e l y b y m e a n s of a coeliaeo- g raphy . Before, dur ing , a n d a f t e r oral t r e a t m e n t w i t h Diazoxide , b lood glucose levels, s e rum insu l in (IMI) a n d s e r u m nones t s r i f i ed f a t t y acids ( N E F A ) were d e t e r m i n e d u n d e r n o r m a l cond i t ions a n d d u r i n g glucose- a n d tol- b u l t a m i d e to le rance tes ts . T he resu l t s of these obse rva - t ions suggest , in a d d i t i o n to r e su l t s o b t a i n e d in v i t ro w i t h t he i so la ted p a n c r e a s of t h e r abb i t , t h e r a t a n d t h e min ia - t u r e pig, t h a t in su l in seere t ion is i n h i b i t e d b y Diazoxide . No side effects of t h e d r u g on t h e l iver, t h e k idneys , a n d t he h a e m a t o p o e t i e s y s t e m were obse rved . Two p a t i e n t s deve loped a m a r k e d h i r s u t i s m whi le on Diazoxide . Af te r a few days on t h e m i n i m a l effect ive dosage ( 4 - - 5 m g / k g p. o. ) in one p a t i e n t nausea , v o m i t u s a n d s i n u s t a c h y c a r d i a oecured before a re l iab le effect on t h e h y p o g l y c e m i a h a d b e e n no ted . I n t h i s ease, t h e c o m b i n a t i o n of Diazox ide a n d t r i c h l o r m e t h i a z i d e d id no t , as expec ted , p o t e n t i a t e a n d increase t h e effect of Diazoxide , b u t h a d to be dis- e 0 n t i n u e d because of inc reas ing in to le rance .

63. The inf luence of intravenous injection of 1-1eucine (rabbits) on glycogen synthesis by liver slices incubated in vitro. MAal)~ DEL CARMEN GAVel& FERNANDEZ and J.L. 1%. CA~DELA. Inst. "H. Maranon" C.S.I.C., Madrid (Spain).

Rabbits of both sexes were injected intravenously with l-leueine (i00 mg/kg) and ten minutes later they were beheaded after a blow on the head. Liver sclices were i n c u b a t e d in H a s t i n g s buffer s u p p l e m e n t e d w i t h glucose (30 raM/l) a n d glucose -UJ~C (0.1 /~c/ml). T he gas phase was 95% O2 : 5% CO2. The results obtained showed that injection of l-leucine: a) raises the glucose uptake of the liver slices; b) stimu- lates "de nero" synthesis of glycogen; c) has no apparent effect on glycogen synthetase, phosphorylase or gluco- kinase.

64. Diabetic neuropathy in animals and man. C. 1~. GILLIS, U n i v e r s i t y of Glasgow, Scot land .

The p e r i p h e r a l n e r v e s f rom t h e a m p u t a t e d lower l imbs of s ix d i abe t i c p a t i e n t s w ho e x h i b i t e d signs a n d s y m p t o m s of n e u r o p a t h y h a v e b e e n carefu l ly s t ud i ed his to logical ly . T h e t o t a l n u m b e r of n e r v e fibres a n d t h e i r d i a m e t e r h a s b e e n assessed a t va r ious co r r e spond ing levels of t h e femo- ra l a n d sciat ic axes a n d h i s t o g r a m s of t h e n e r v e f ibre d i a m e t e r spec t r a a t t hese va r ious levels h a v e b e e n con- s t ruc t ed . The resu l t s show t h a t t h e r e appea r s to be a specific pa t - t e r n of n e r v e f ibre d e g e n e r a t i o n in d i abe t e s mel l i tus . This is emphas i s ed b y c o m p a r i s o n w i t h a s t u d y of t h e n e r v e f ibre p o p u l a t i o n s of t h e leg in chron ic oeelusive a r t e r i a l d isease in m a n (GAI~VEN, GAIRNS a n d S~ITI~, 1962). ~ 1 18 m o n t h - o l d co lony of u n t r e a t e d a l l oxan d iabe t i c r a t s ha s b e e n s t ud i ed in a s imi la r m a n n e r a n d a t t h i s s tage i t appea r s t h a t t h e r e are m a n y s imi lar i t ies to t h e p a t t e r n of n e r v e f ibre d e g e n e r a t i o n obse rved in t h e h u m a n pa- t i en t s . The signif icance of t h e resu l t s a n d t h e m e t h o d s b y w h i c h t h e y were o b t a i n e d is discussed.

65. Serum insul in- l ike activity measured by the isolated fat cell method. g. GI~IEMAlgN. I n s t . Of Med. Phys io logy , Copenhagen .

I s o l a t e d ad ipose ceils were i n c u b a t e d in I(rebs-1%inger b i c a r b o n a t e buffer w i t h 0.1 m g g lucose /ml a n d a l b u m i n 1%. Insu l in , in c o n c e n t r a t i o n s of a b o u t 2 0 - - 3 0 ~ U / m l

or more , s t i m u l a t e d t h e convers ion of glucose 1-14C tO ~CO~ 1 5 - - 3 0 fold. The m i n i m u m effect ive c o n c e n t r a t i o n was a b o u t 0.25 #U/ml . W h e n t h e glucose c o n c e n t r a t i o n of t h e m e d i u m was increased to 2 m g / m l t h e m a x i m a l response to insu l in was a 3 - - 4 fold increase in glucose convers ion . The sens i t iv i ty , however , r e m a i n e d t h e same. Sera were t a k e n f rom 3 n o r m a l sub jec t s d u r i n g a n oral glucose to le rance tes t . The se ts , a d j u s t e d to a glucose c o n c e n t r a t i o n of 2 mg/ml , showed " I n s u l i n - l i k e " effects of more t h a n 30 #U/ml . A d d i t i o n of insu l in a n t i s e r u m h a d no effect. F a s t i n g sera, d i lu ted to 5 % in buffer , showed a " suppress ib le I L A " of 2 0 - - 4 0 # U / m l (measured " I L A " t imes d i l u t i on factor) . I n s u l i n (2,5 a n d 5 #U/ml ) a d d e d to t h e sera was q u a n t i t a t i v e l y recovered . 3 0 - - 6 0 m i n u t e s a f t e r t he glucose load " t h e suppress ib le I L A " h a d inc reased to 100- -160 /~U/ml. Af te r 120 m i n u t e s i t was 20 /~U/ml or less. I n all sera " n o n suppress ib le I L A " equivalent to 40--i00 /~U/ml was found. This fraction was constant throughout the glucose tolerance test for each individual.

66. Fine structure of B-cel l from pancreas slices incubated in vitro. J. GONEz-AcEBO, C. LOP]~z-QuIJADA and J.L.R.-CAN- DELA. Institute "G. Maranon" C.S.I.C., Madrid, Espagne.

Slices of the tail of the rabbit pancreas were incubated in a shaking incubator with Krebs-Henseleit medium supplemented with glucose (0.6 mg/ml) pyruvate, fu- marate and glutamate (5 mM), with a gas phase 95% 0~: 5% CO 2 and at 38~ There were no significant changes in t h e fine s t r u c t u r e of t h e islet ceils a t t r i b u t a b l e to t h e i ncuba t ion . Combined g l u t a r a l d e h y d e a n d o s m i u m fixa- t i on gave good p r e s e r v a t i o n of t h e sec re to ry p r o d u c t s of t h e B-cells. The m e c h a n i s m s of d e g r a n u l a t i o n a n d g ranu- l a t i on of t h e B-cells were obse rved to be t h e same as those r e p o r t e d for t he a n i m a l " i n v i v o " . The i n t e n s i t y of secre t ion va r i ed if t h e an ima l s were ki l led a f t e r be ing knocked unconsc ious or a f t e r be ing in jec ted w i t h a l e tha l dose of aneas the t i e . B e t a cell secre t ion was more p romi - n e n t in t h e fo rmer ease. T h e A-cells in genera l r e m a i n e d s tab le du r ing t h e i n c u b a t i o n t ime. The m e t h o d is con- s idered to be va l id for f u r t h e r s tud ies on t h e m e c h a n i s m of g ranu le release a n d syn thes i s in t h e p a n c r e a s of t h e r abb i t . P r e l i m i n a r y f indings conce rn ing morpho log iea l changes of t h e islet cells a f te r glucose s t i m u l a t i o n in v i t ro , will be p resen ted . E v i d e n c e of a doub le m e m b r a n e l im- i t ing t h e b e t a g ranu le will be shown.

67. Studies on the diabetogenic activity of an antibiotic: streptozotocin. ALAIlg E. GONET a n d ALBERT E. I~ENOLD. I n s t i t u t de B ioch imie Clinique, Geneva , Swi tzer land .

Streptozotocin is a large-spectrum antibiotic from strep- tomyees achromogenes with the empirical formula C1411~7N~012. Strepbozotoein preparations have been reported to exhibit, in addition, antitumoral and diabeto- genie activity in several species (RAI~IE~rEN etal. : Cancer Chemother, Rep. 29, 91 (1963); EVANS et al.: Cancer Chemother. Rep. in press). This unusual combination of biological activities led us to a more complete study of t h e d iabe togen ie ac t ion of t h e c o m p o u n d . A d i abe t i c s t a t e was easi ly i nduced in well over 75O/o of n o n - f a s t e d a lb ino W i s t a r r a t s in j ec ted once w i t h 65 m g / k g i .v . of a s t r ep to - zo toe in p r e p a r a t i o n o b t a i n e d f rom t h e U p j o h n Co. The re were no ear ly dea ths . Mild h y p e r g l y c e m i a a p p e a r e d w i t h i n hou r s a n d t h e n progressed g radua l ly to m a x i m a l non- f a s t ing levels b e t w e e n 400 a n d 1000 nag pe r 100 ml a f t e r 3 weeks. P o l y u r i a r e ached 5 0 - - 1 0 0 ml pe r d a y w i t h up to 15% glucosur ia . L i p e m i a was seen in severa l in s t ances ,

152 O r g a n i z a t i o n Sec t ion �9 A b s t r a c t s Diabetologia

although there was no weight loss or ketonuria and a 24 hour fast reduced blood glucose levels to the range of 120--200 mg per i00 ml. There was rapid (days) de- granulation of the fl cells of the pancreatic islets with hydropie degeneration and accumulation of glycogen. Pancreatic insulin content decreased from 2.5--4.5 U/gin to 0.1--0.4 U/gin. The most striking histological observation on the endocrine pancreas, however, was that of a remarkable proliferation of cells which exhibited the characteristics of ~ cells with aldehyde-thionin. Thus, the diabetic state produced by streptozotoein differs markedly from that produced by fl-eytotoxic agents and clearly warrants further study.

68. Diab~te exp6rimental chez le rat par injections de glucagon ou ingestions d'huUe. L. GEASSI e t J . P . FELBER. Clinique M@dicale Unive r s i - ta i re , L a u s a n n e .

U n lo t de 53 ra ts , divis~s en 5 groupes , a sub i d u r a n t u n e p@riode de 7 k 10 jours , k r a i son de 3 doses p a r jour , les traitements s u i v a n t : 1. g lueagon, in jec t ions s .e . (1 rag / j /200 g) 2. hui le d 'o l ive pe r os (3 • 1 ml) 3. so lu t ion glueosge pe r os (3 • 1 ml , 3 g/kg) 4. insul ine, in jec t ions s.c. (dose c o r r e s p o n d a n t k la dose c o n t e n u e d a n s le g lucagon) 5. NaC1, in jec t ions s.c. U n e @preuve de to le rance au glucose, pe r os (3 g/kg), a ~t@ effectuge chez tousles rats, avant et apr@s le traite- ment. Par rapport aux rats t@moins -- ayant regu du glucose, de l'insuline ou du NaCl -- nous observons ehez les rats trait@s au glucagon ou ~ l'huile d'olive: i. une perte de poids ehez les premiers, une prise de poids ehez les seconds; 2. une glyc@mie ~ jeun non modifi~e, mais une diminution signifieative de la tol@ranee au glucose aecompagn@e dans la majorit@ des cas d'une diminution relative ou absolue de l'insulin@mie immuno-reactive ; 3. u n e r@serve p a n e r ~ a t i q u e en insu l ine n o n diminu@e, d~ te rminee p o s t - m o r t e m apr~s e x t r a c t i o n ~ l 'g thanol-HC1 du pancr@as. L ' i n t e r e t de ces r@sultats cons is te en l ' ana log ie qu ' i l s p r e s e n t e n t avee ce r ta ines fo rmes de diab@tes cl iniques.

Experimental diabetes in the rat by glucagon injection or oil ingestion. L. GRASSI and J.P. FELBER. Univ. Med. Clin., Lausanne, Switzerland.

A batch of 53 rats, divided into 5 groups, was subjected to the following forms of treatment for a period of 7-- i0 days, 3 doses being administered daily: i. subcutaneous injections of glueagon (i rag/200 g per diem); 2. oral olive oil (3 • I ml) ; 3. oral glucose solution (3 • 1 ml, 3 g/kg) ; 4. subcutaneous injections of insulin (dose corresponding to that of the glueagon); 5. subcutaneous injections of NaCl. A glucose tolerance test (3 g/kg orally) was carried out oh all the rats before and after treatment. As compared with the control rats -- those which received �9 glucose, insulin or NaCl -- the following findings were o b t a i n e d w i t h t h e r a t s t r e a t e d w i t h g lucagon or olive oil: 1. t h e g lueagon r a t s los t weight , t h e r a t s on olive oil ga ined we igh t ; 2. t h e r a t s on g lucagon or olive oil h a d a n u n c h a n g e d f a s t i ng b lood sugar level, b u t showed a sig- n i f i can t decrease in glucose to le rance , a c c o m p a n i e d in m o s t cases b y a n abso lu t e or r e l a t ive fall in t h e i m m u - no reac t ive insu l in b lood leve l ; 3. in t h e r a t s on g lucagon or olive oil t h e p a n c r e a t i c insu l in reserve was n o t reduced , as d e t e r m i n e d pos t m o r t e m b y e thanol -HC1 e x t r a c t i o n of t h e pancreas .

These f indings are of i n t e r e s t because of t h e ana logy t h e y p r e sen t w i t h t hose in c e r t a i n fo rms of cl inical d iabe tes .

69. The influence of the duration of diabetes and the metabolic state on motor conduction velocity, electromyography and biot hesiometry. G. GI~EGEI~SEN. The 2nd Clinic of I n t e r n a l Med. a n d t h e Dep. of Neurology, K o m m u n e h o s p i t a l e t , Aa rhus .

A n i n v e s t i g a t i o n on 145 d iabe t i cs r evea led a close cor- r e l a t i on b e t w e e n t h e a b o v e m e n t i o n e d p a r a m e t e r s a n d t h e d u r a t i o n of d i abe t e s in y o u n g people. I~owever , a decl ine in n e r v e f u n c t i o n was f o u n d also in p a t i e n t s w i t h less t h a n 5 yea r s ' d u r a t i o n of d iabe tes . A t t e m p t s to de- m o n s t r a t e a m e t a b o l i c inf luence on n e r v e func t i on will be p r e s e n t e d .

70. Effect of "Dicumarol" (Bishydroxycoumarin) on the insulin secretion in vitro. MA. ANTONI~ Gi~IVTER, CLEMENTE LOPEZ-QuIJADA and J.L. I~-CANDELA. Institute "G. Maranon" C.S.I.C., Ma- drid, Spain.

In a large group of experiments which will be published elsewhere we have studied the effects of the uncouplers of oxidative phosphorylation on the secretion of insulin in vitro. The unexpected results obtained with Dicumarol, up to the present time unpublished, are the subject of this communication. Insulin was extracted from the incubation medium by the Grodsky and Tarver method, and determined either with the fat pad by increased 14CO2 production (RENOLD), or b y t h e immuno log i ca l m e t h o d (HALES a n d ]:~ANDLE). The resu l t s o b t a i n e d in t h e d i f ferent series of e x p e r i m e n t s show t h a t D i c u m a r o l s t imu la t e s t he secre t ion of insu l in a n d t h a t t h i s s t imu lus seems i n d e p e n d e n t of t h e concen- t r a t i o n of glucose in t h e m e d i u m .

71. Zur Frage der Ausscheidung und des Wirkspiegels von Nt-n Butylbiguanid. H. HALLEI~ und S.E. STI~AUZENBEI~G. Med. Klin. u. Poli- klin., Med. Akad. Dresden, Germany.

Mittels eines Papierehromatographisehen Verfahrens wurde die Ausseheidung des Butylbiguanides im I-Iarn gepriift, l~aeh einmaliger Belastung mit 200 mg wurden in 8 Stunden 75% und innerhalb 24 Stunden 85--90% der Substanz unver/inderb im Earn gefunden. Die Halb- wertszeit betrug zirka 2.5 Stunden. Unter der Voraus- setzung einer totalen Clearance erreehnen sich Plasma- konzentrationen unter 1.0/~g/ml. Die sieh ffir die Klinik ergebenden praktisehen Folgerungen sowie die gleichzeitig mit erfalBten Eliminationsverh~Itnisse yon Kreatinin und Kreatin werden diskutiert.

The excretion and effective plasma concentration of N'-n- Butylbiguanide. H. HALLER a n d S .E . STRAUZENBERG. Medical Clinic a n d Pol ic l inie of t h e Carl G u s t a v Carus Medical A c a d e m y , Dresden .

T h e exc re t ion of b u t y l b i g u a n i d e in t h e u r ine was de ter - m i n e d b y m e a n s of p a p e r c h r o m a t o g r a p h y . Af te r admi- n i s t r a t i o n of a single dose of 200 m g of t h e c o m p o u n d , 7 5 % was exc re t ed u n c h a n g e d in t h e u r ine in 8 h o u r s a n d 8 5 - - 9 0 % w i t h i n 24 hours . The t i m e r equ i r ed for 5 0 % e l im ina t i on was a p p r o x i m a t e l y 2.5 hours . A s s u m i n g 100% clearance , t h e ca lcu la ted p l a s m a c o n c e n t r a t i o n s are below 1.0 #g/ml . The impl i ca t ions for cl inical p rac t i ce are d iscussed as well as t h e r a t e s of e l im ina t i on of e rea t ine a n d e rea t in ine , w h i c h were d e t e r m i n e d a t t h e same t ime .

Vol. 2, _No. 2, 1966 Organization Section �9 Abstracts 153

72. Anderungen im Eiweifl und Fettstoffwechsel bei oraler und Insulin-Behandlung yon Zuckerkranken.

T. HALlos, B. LOCZKA, F. S~A~o~r E. MA~OR. Abtei- lung fiir innere Medizin I. ganos Krankenhaus, Budapest, Ungarn.

Zahlreichc Zuekerkranke leiden auch an St6rungen des l~ettstoffwechsels. Einlge Autoren sind der Ansicht, daI~ die Zuckerkrankheit primer eine FettstoffwechselstSrung sei, und die Manifestation des Hyperglykemiesyndroms erst durch die St6rung des Lipoidstoffwechsels -- infolge pathologiseher Funktion von Enzymen -- herbeigefiihrt werde. Auf Grund unserer bisherigen Ergebnisse m6chten wir darauf hinweisen, dal] durch die lqormalisierung des Zuckerhaushaltes, z.B. durch orale Antidiabetika, nur ein Teil der komplexen Stoffwechselst6rung eliminiert wird. Gleichzeitig verschlimmert sich die St5rung des Fett- bzw. EiweiI~-Stoffwechsels.

Changes in protein and fat metabolism during oral antidiabetic and insulin therapy of diabetics.

T. HAL~rOS, B. LOCZKA, F. SALA~O~ and E. MAzO~. First Department of Internal Medicine of the Janos Hospital, Budapest.

Many diabetics suffer from more or less serious disturbances in lipid metabolism. Some authors are of the opinion that diabetes is primarily a disturbance of fat metabolism and that the hyperglycaemic syndrome becomes manifest only as the result of the disturbance of lipid metabolism due to the pathological functioning of enzymes. Our results so far seem to indicate that normalization of sugar metabolism, e.g., by treatment with oral antidiabetics, eliminates only a part of the complex of metabolic disorders, and that the disturbances in lipid or protein metabolism are intensified.

73. Specific uptake in the islet tissue of the diabetogenic agents alloxan and dehydroascorbic acid.

L. HA~YimaSTR6~, Department of Pharmacology, Royal Veterinary College, Stockholm 50, Sweden.

The distribution in mice and young rats of L-ascorbic acid-l-laC and dehydro-L-ascorbic acid-l-saC has been studied by means of whole-body autoradiography and microautoradiography. These studies have shown that dehydroascorbic acid immediately after intravenous injection accumulated in the pancreatic islets of the adult mice. The concentration of aseorbie acid was not observable in the islets until 4 hours after injection. Microauto- radiography showed that most of the islet cells were labelled; possibly all of them. The concentration in the exocrine pancreas was low during the whole investigation period. In the young rats which are known to be restistant to the diabetogenic action of alloxan neither dehydroascorbic acid nor ascorbic acid was accumulated in the pancreatic islets. In a similar study (together with Dr Ullberg) alloxan-2- sac was injected to adult mice. The radioactivity was rapidly accumulated in the pancreatic islets and most of the islet cells were labelled. The concentration in the islets seemed to be the highest in the body. I t persisted longer in the pancreatic islets than in most other tissues, and was still observable 24 hours after injection. The distribution of the oxidized and reduced non-diabetogenic conversion products of alloxan has also been studied (together with Drs Hellman and Ullberg). No accumulation in the pancreatic islets was observed after injection of these non-diabetogenic compounds. I t was also shown that alloxan did not accumulate in the islet cells of young mice and rats.

74. Das Verhalten yon Fett- und Kohlehydratstoffwechsel- Metaboliten bei Gesunden und Diabetikern nach parcntaler Applikation versehiedener Zncker. H. HAMME~L, O. PIC~LE~, M. S~UDLA~. I. Medizinische .A.bteilung des Wilhelminspitals der Stadt Wien, Wien, Osterreich.

Bei Diabetikern, Pa t i en ten mit einem positiven Tolbuta- midtest, bzw. gesunden Personen wurden vor der iv. Applikation yon 50 g Dextrose bzw. Laevulose (200 ml einer 25%igen L6stmg, Einlaufgeschwindigkeit 5 Minu- ten), sowie 30, 60, 120 und 180 Minuten nach Unter- suehungsbeginn folgende Bcfunde erhoben: Blutzucker (Glucose enzymatisch), Laktat , Pyruvat , freies Glyerin, Gesamtglycerin und Triglyeeride (enzymatisch), freie Fett- s~iuren (nach DOLE und MEI~Et~TZ). Die Belastungen er- folgten nach einer 14stiindigen Nahrungskarenz der Pro- banden unter Grundumsatzbedingungen. Die Ver~nderungen der angefiihrten Parameter nach den Zuckerbela- stungen sind sowohl hinsichtlich der Intensi ta t als auch der Dauer yon der jeweiligen Stoffwechsellage abh~ngig. Die dargelegten Ergebnisse und die daraus ableitbaren Konsequenzen ftir Theorie und Klinik werden diskutiert.

The behaviour of fat and carbohydrate metabolites in healthy persons and in diabetics after parenteral administration of various sugars. H. I-IAM~ERL, 0. PICttLEI~ and M. S~UDLA~. First Medical Departmen~ of the Wilhelmina Hospital, Vienna.

Diabetics, patients with a positive tolbutamide test, and healthy persons were given 50 g of either dextrose or laevulose (200 ml of a 25% solution) administered by intravenous injection over a period of 5 minutes. Before the injection and at intervals of 30, 60, 120 and 180 minutes afterwards determinations were made of the blood levels of glucose (enzyme method), lactate, pyruvate, free glycerol, total glycerol and triglycerides (enzyme method), and free fat ty acids (DOLE and MEINE~Z). The injections were made after the patients had fasted for 14 hours under basal metabolic conditions. Both the intensity and the duration of the subsequent changes in the blood levels of the various metabolites were dependent on the metabolic state at the time of the injection. The results obtained and their thedretical and clinical implications are discussed.

75. Stoffwechselwirkungen von 3.5-Dimethylisoxazol im Hunger und bei Diabetes. A. I'I~SSELBLAT% P. BUBENttEIiVIEE und U. SCHWABE. Pharmakologisches Ins t i tu t der Universit~t G6ttingen.

3.5-Dimethylisoxazol hemmt die Lipolyse im Fettgewebe und senkt die Konzentrat ion der unveresterten Fettsau- ren im Blur. Diese Lipolysehemmung hat besonders im Hunger und beim akuten Diabetes, we sonst vermehrt Fetts~uren mobilisiert werden, Auswirkungen auf den Stoffwechsel. So wird an hungernden Rat ten und bei l%atten, bei denen dureh Anti-Insulinserum ein akuter Insulinmangel ausgel6st wurde, die Bildung yon Keton- k6rpern in der Lcber unterdriiekt und die Ketons~ure aufgehoben. In seiner starken antiketogenen Wirkung und der Lipolysehemmung zeigt 3.5-Dimethylisoxazol Parallelen zur Wirkung yon Insulin. Im Gegensatz zu Insulin vermag es jedoch die Glucoseutilisation nu t un- wesentlich zu steigern. Da der Ausfall yon Fetts~uren und Ketok6rpern als Energietr~ger im Hunger und bei aku- tern Diabetes hier nicht durch einen gesteigerten Glucose- stoffwechsel ausgeglichen werden kann, fiihrt 3.5-Dime- thylisoxazol zu einem Anstieg der Gluconeogenese. An hungernden Rat ten steig~ der Einbau yon Glycin=C 1~ in Glucose und Glykogen, die Corticosteronkonzentration im Plasma und die Stickstoffausscheidung im Urin. Diese Stoffwechselwirkung des 3.5-Dimethylisoxazol steht im

154 Organization Section Abstracts Diabotologia

Gegensatz zu der bekannten anabolen }Virkung yon In- sulin.

Metabolic effects of 3.5-dimethylisoxazole in hunger and in diabetes. A. I'IASSELBLAT~, P. BUBENHEIM~.~ and U. SCRWABE. Pharmacological Inst i tute of the University, G6ttingen.

3.5-Dimethylisoxazole inhibits ]ipolysis in adipose tissue and lowers the concentration of free fa t ty acids in the blood. This inibition of lipolysis produces alterations in metabolism, particularly in states of hunger and in acute diabetes, that are normally characterized by increased mobilization of fat ty acids. Thus, in starving rats and in rats made insulin-deficient by the administration of anti- insulin serum, the formation of ketone bodies in the liver was suppressed and the ketonaemia was eliminated. In its antiketogenic action and in its inhibiting effect on lipolysis, 3.5-Dimethylisoxazole shows some parallels to the action of insulin. In contrast to insulin, however, it causes only an insignificant rise in glucose utilization. Since in states of hunger and in acute diabetes, the absence of fat ty acids and ketone bodies as sources of energy cannot be compensated for by increased glucose metabolism, 3.5-Dimethylisoxazole leads to increased formation of glucose. In starving rats, there is an increase in the conversion of laC-glyeine to glucose and glycogen, in the concentration of corticosterone in the plasma, and in the excretion of nitrogen in the urine. This metabolic effect of 3.5-Dimethylisoxazole contrasts with the well- known anabolic action of insulin.

76. Cobalt and islet tissue. N. HAvu, E. PIItL and S. FALKMEtr Inst i tute of Patholo- gy, University of Umes Ume& 6, Sweden.

Preceding studies disclosed a marked cobalt-concentrating ability of islet tissue, apparently confined to the ul-(D-) cells and/or the fl-cells. By means of some recently developed methods for glutathione (GSH) assay of islet tissue and ultrastruetural localization of heavy metals we have now studied some biochemical effects of cobalt administration and tried to improve the morphological analysis. Using the isolated islet tissue of the marine teleost Cottus scorpius, it was found that a single i.m. injection of CoCI~ at L.D. 50 evoked a significant decrease in the GSH content of the islets at the same observation times as when the islet cell accumulation of 5sCo is at its maximum (after 2 d). The pat tern of this islet GSH decrease thus differed markedly from the more rapid GSI-I lowering effect of a diabetogenic dose of alloxan (max. after 2 h). l~epeated i.m. injections of CoC12 at L.D. 50 evoked clear- cut necroses of the parenchymal c~lls in that region of the islets where the metal ions are known to accumulate. These islet changes were accompanied by both hypo- and hyperglycemic reactions. The ultrastrueturM investigations, still in progress, have so far shown signs of metal accumulation and early de- generative lesions in some of the organelles of the islet parenehymal cells.

77. The metabolism of mammal ian pancreatic islets studied with the cartesian diver micro-gasometer. C. I.IELLEI~STIa61VL Histological Department, University of Uppsala, Uppsala, Sweden.

In studies of the metabolism of mammalian B-cells, Cartesian divers were used for analysis of the oxygen consumption of isolated and surviving pancreatic islets from obese-hyperglycemic mice. When single islets (dry weight 1--10 #g) were incubated in Krebs-l%inger phos-

phate buffer at + 37 ~ C, the respiratory rate was constant for several hours. A striking elevation of the respiratory rate was observed when the medium was supplemented with glucose; a less pronounced stimulation of the oxygen uptake being obtained with mannose and fructose. Galactose, 2-deoxy-glueose and marmoheptulose were without effect on the endogenous respiration. However, addition of mannoheptulose blocked the stimulatory effect of glucose on the islet respiratory rate. The results are considered representative for the B-cells since in obese-hyperglycemic mice this cell type comprises more than 90% of the isIet tissue.

78. Lactate dehydrogenase isoenzymes in mammal ian B-cells. B. I-IEL~,~ASr and I.-B. TXLJEDAL. Departments of Histo- logy, Karolinska Instituter, Stockholm 60, and University of Umeg~, Ume&, Sweden.

Disc eleetrophoresis, microehemieal assays and histo- chemical staining were employed in a characterization of the lactate dehydrogenase (LDH) a c t i v i t y in the pancreatic islets of obese-hyperglycemic mice. Five L D H isoenzymes were eleetrophoretically separated, 65% of the total enzyme activity being represented by LDH-5. A theoretical calculation, based on the sub-unit theory, revealed a proportion of 14% H- and 86% M-units in the LDH of the pancreatic islets. The islets displayed significantly lower LDH act ivi ty than the exoerine paren- ehyma. A considerably lower ratio between the dehydrogenase activities towards 2-oxobutyrate and p.yruvate was observed in both the endocrine and exocrme pancreas as compared to the heart. By means of a histo- chemical substrate film technique, a weaker staining for LDH activity was noted in the islet B-cells than in the acinar tissue. Addition of 0.08 mM phenazine methosul- phate did not apparently influence the degree of staining either in the endocrine or exocrine pancreas. The present data suggest that the islet B-cells are well equipped for the accumulation of lactate and the establish- ment of an oxygen debt.

79. Influence of dietary fats and carbo-hydrates on lipids and lipoproteins in diabetic retinopathy. A.J . I-Iou~s~VLLE~. Dept of the Eye-Clinic, l~otterdam.

In a previous study it was found that patients with diabetic ret inopathy had moderate to severe hyperlipemia. A study was undertaken to establish the influence of saturated and poly-unsaturated fats on the lipids in diabetic retinopathy. I t happened that in most of the cases the serum lipids rapidly improved after poly-unsaturated fat-intake. In a few cases there was either no improvement or even deterioration. I t was furthermore shown, that carbohydrates had a marked influence on the serum lipids, and of them especially the mono- and disaecharides (su- crose). The ~-fl-lipoproteins reflected the changes in diet, with the formation of a large pre fi-lipoprotein fraction during carbohydrate intake. This fraction disappeared after intravenous heparin, as could be demonstrated with ~/new agar-electrophoretic technique. After changing the diet from carbohydrate-rich;into fat-rich, the carbohydrate induced pre-fi-lipoprotein disappeared often only after 2--3 weeks. During these studies a hitherto unknown lipoprotein fraction was found in the plasma of several cases of diabetic retinopathy.

80. Bakteriurie und Diabetes. S. Iv.cA, C. DODICA und Z. IovAzcovlei. Klinisch-chemi- sches Laboratorium des Kreiskrankenhauses Lugoj, Lu- goj, gum~nien.

Vol. 2, No. 2, 1966 Organiza t ion Sect ion �9 Abst rac ts 155

f3ber die t t / iuf igkei t der Bakter iur ie im Diabetes sind die Meinungen verschieden. Es werden fiber h6here aber ebcnso h/iufig aueh tiber normale Anfa l l squoten ber iehte t . Unsere Un te r suchungen ers t recken sieh auf 106 Diabet i - ker, davon 21 Jugend l i che zwisehen 15 und 32 J a h r e n und 85 P a t i e n t e n zwischen 48 und 66 Jahren . Azetonur ie zeigten 17 jugendl iehe und l l d e r / i l t e r e n Pa t ien ten . Als Kont ro l le wurden zwei Gruppen gesunder Personen der- selben Altersklassen untersucht . Bet allen wurde die Urin- keimz/ihlung in Abs tgnden yon drei Tagen, zwei Z/ihlun- gen t/igtieh, insgesamt 6mat durehgeffihrt . Es wurde die Mi t te l s t rah l technik angewandt . Fi i r die zwei am gleichen Tag ausgeff ihrten Z~hlungen wurde die erste Probe dem !VIorgenurin, die zweite dem I-Iarn nach sechssti indiger Arbei t en tnommen. Ergebnisse : Bet 5 % der Jugend l i chen wurden im Morgen- urin, und bet 10~ im I-Iarn nach sechssti indiger Arbeits- be las tung pathologische Ke imzah len gefunden. I n ke inem dieser Fgl le bes tand eine Azetonurie . I n der Kontrol l - gruppe be t rug der Prozentsa tz im Morgenurin 4.5 %, naeh Arbe i t 6.5%. Bet den /il teren Diabe t ike rn fanden wir einen pathologischen Befund in 6% der F//lle im Morgen- harn und in 18.7% im H a m nach Arbei tsbelas tung. Auch un te r diesen befanden sich F/ille mi t Azetonurie . Bet der /i l teren Kont ro l lg ruppe fanden wir bet 60/o einen pa tho- logischen Ausfall im lVforgenurin a n d bet 7 .3% im H a m nach Arbei tsbelas tung. Es ergibt sich also eine bemerkenswer te Ste igerung der Anfal l squote im I-Iarn nach Arbeits- be las tung in der Gruppe der g l teren Diabet iker . Ebenso bemerkenswer t erseheint die Tatsache, dal3 sich Pat ten- t en mi t Azetonur ie den t t a rnwegs in fek ten gegenfiber rest- s touter erweisen. Die Diabe t iker h6herer Altersklassen scheinen ffir I-Iarnwegsinfekte empf/inglieher zu sein.

B a c t e r i u r i a a n d d iabe te s .

�9 S. ILCA, C. DODIOA and Z. IOANOVlCI. Clinico-chemieal Labo ra to ry of the Municipal Hospi ta l , Lugoj , Ruman ia .

Opinions differ as to the f requency of bacter iur ia in diabetes. Increased rates have been reported, bu t there are equal ly numerous reports of normal rates. Our invest i- gat ions cover 106 diabetics, of w h o m 21 were be tween the ages of 15 and 32 years, and 85 were in the age group 48--66. Aeetonur ia was present in 17 persons in the younger group and in 11 of those in the older group. Two groups of hea l thy persons in the same age groups served as controls. I n all subjects, two bacter ia l counts per day were per formed a t three day- in terva ls up to a to ta l of d counts. The first of the dai ly counts was made on the morn ing urine, the second af ter the subject had per formed six hours ' work. Resul t s : of the 21 diabetics in the younger age group, 5 ~ showed a pathological bacter ia l count in the morn ing urine and 10% in the ur ine af ter six hours ' work. None of the pa t ien ts wi th pathological counts exhib i ted ace- tonuria . I n the control group of younger subjects, pa tho- logical counts were found in the morn ing ur ine in 4 .5% and in the ur ine af ter work in 6 .5% of the subjects. I n the diabet ic pa t ien ts of the older age group, the corresponding figures were 6% and 18.7%. Again, none of the pa t ien ts wi th pathological counts had aeetonuria . I n the control group of older subjects, 6% showed a pa tho- logical count in the morn ing urine and 7.3% in the ur ine af ter work. Thus, there was a marked increase in the percentage of pathological counts in the ur ine af ter work among the diabetics in the older age group. I t is also remarkab le t h a t the pa t ien ts wi th aee tonur ia were more res is tant to infect ion of the u r ina ry t ract . Thus, diabetics in the upper age groups appear to be more liable to ur inary t r ac t infections.

Diabetologia, Vo]. 2 i1

156 Organization Section Diabetologia

M i n k o w s k i A w a r d 1967 - - Pr ix M i n k o w s k i 1967 - - M i n k o w s k i - P r e i s 1967

The European Association for the Study of Diabetes announces the 1967 Minkowski Award donated by Farb- werke Hoeehsb AG. Nominations are requested and should be in the hands of the Secretary of the Association before January 15th, 1967. Nominations may only be made by members of the European Association for the Study of Diabetes.

The Award consists of a certificate and of DM 10 0 0 0 . - plus travel expenses. I t is awarded for distinetion manifested by publications which cdntribute to the advance- ment of knowledge concerning diabetes mellitus. Candi- dates should be less than 40 years of age on January 1st, 1967. The Award will be given in relation to research, which has been carried out in Europe by a person nor- really resident in Europe.

Nominations should be sent to the Secretary of the Association, Prof. ALBE~T E. I~ENOLD, Inst i tut de Bio- ehimie Clinique, Sentier de la Roseraie, 1211 Geneva (Switzerland). I t is to be in the form of 6 copies of a curriculum vitae of the candidate, 6 copies of each of the relevant pubIications and 6 copies of a statement by the proposer(s) in English, French or German, of the manner in which the candidate's published contributions have aided knowledge concerning diabetes mellitus. Such a statement should not exceed 100 words.

The recipient of the Award will be expected to give a lecture on the relevant research at the Sixth Congress of the International Diabetes Federation, Ju ly 30 to August 4, I967, in Stockholm, Sweden. He will be expected to write an account of the research for publication in Diabetologia, the Journal of the Association.

L'Assoeiation Europdenne pour l 'E tude du Diab@te annonee la raise en contours du Prix Minkowski 1967, institu6 par Farbwerke tIoechst AG. Les propositions peuvent ~tre faites par un ou plusieurs membres de l'As- soeiation et devront parvenir avant le ler Janvier, 1967, au Secrdtaire: Prof. ALBERT E. I:~ENOLD, Inst i tut de Biochimie Clinique, Sentier de la Roseraie, 1211 Gen@ve 4, (Suisse).

Le Pr ix consiste en un dipl6me et une somme de 10000 DM; les frais de voyage du laur6at seront rem- bours6s. Le Prix est destin6 N r6eompenser des publications ayant 6t6 effeetuges en Europe et ayant permis l 'avaneement des connaissanees dans le domaine du diabgte suer6. Le laur@at devra gtre ggg de moins de 40 ans au ler janvier 1967 et r6sider habituellement en Europe.

Chaque proposition devra eomporter: un curriculum vitae du candidat en 6 exemplaires, 6 exemplaires des publications correspondantes et 6 copies d 'une appre- ciation personnelle du ou des membres prgsentant le candidat (en anglais, en franqais ou en allemand), appre- ciation qui portera sur la mani~re dont le travail et les publications du eandidat contribuent ~ l 'avaneement des cormaissanees sur le diab~te sucr@. Cette appr6eiation ne devra pas d@asser 1000 mots.

Le laur@at sera appel@ k donner une conf@rence sur ses t ravaux lors du Sixi6me Congr~s de la F~dgration Internationale du Diab@te, du 30 juillet au 4 aofit, 1967,

Stockholm, Suede. On lui demandera, 6galement, de r@diger un article t ra i tant de ses recherches, qui sera publi6 dans Diabetologia, Journal de l'Association.

Die Europ/iisehe Gesellschaft fiir Diabetologie erbit tet Vorsehli~ge fiir den Minkowski-Preis 1967, yon den Farb- werken Hoeehst AG, gestiftet. Die Vorsehl~ge sind vor dem 15. Januar 1967 bei dem Sekret~r der Gesellschaft, Professor AL~EaT E. RE~OLD, Inst i tut de Bioehimie Clinique, Sentier de la Roseraie, 1211 Gen~ve 4 (Switzer- land), einzureiehen.

Der Preis bestehb aus einer Urktmde sowie aus einer Geldsumme yon 10 000 DM. Die Reisekosten werden rfick- ersbattet. Der Preis belohnt besonders verdienstvolle Publikationen auf dem Gebiet des Diabetes mellitus. Der Empf/~nger soll am 1. Januar 1967 das Alter von 40 Jahren noeh nichb erreicht haben und soll seinen Wohnsitz in Europa haben.

Zu Vorschlggen bereehtigt sind alle Mitglieder der Europaischen Gesellschaft ffir Diabetologie.

Jedem Vorschlag sollen folgende Unterlagen beigefiigb werden: 6 Exemplare eines Lebenslaufes des Kandidaten, 6 Exemplare aller betreffenden Publikationen und6 Exem- plare einer Darstellung des oder der Vorschlagenden in englischer, franz6siseher oder deutscher Sprache, die den Beitrag der Publikationen des Kandidaten an die F6rde- rung des Wissens fiber den Diabetes mellitus darlegt. Die Darstellung soll 1000 Worte nieht tibersehreiten.

Es wird vom Empfgnger des Preises erwartet, dal3 er wghrend des seehsten Kongresses der Internationalen Dia- betes F6deration, vom 30. Jul i bis zum 4. August 1967, in Stockholm, Schweden, einen Vorbrag fiber seine For- schungsarbeiten h/ilt und da2 er eine Darstellung seiner Arbeiten zur Puhlikation in Diabetologia, dem Organ der Gesellschaft, einreichb.

Responsible for the text : Prof. Dr. K. OBERDISSE, 4 Dfisseldorf, 2. Med. Klinik und Poliklinik der Universitat. Responsible for advertisements : EDG~ SEIDLEE, 1 Berlin 31 (Wilmersdorf), tteidelberger Platz 3. Springer-Verlag, Berlin, Heidelberg, New York.

Printed in Germany by Buehdruekerei Georg Appl, Wemding]Schwaben.

Second Annual Meeting of the European Association for the Study of Diabetes abstracts

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