Hormone and Metabolie Research

Editors-in-Chief: R. Levine, Duarte E. F. Pfeiffer, Ulm

Assistant-Editors: R. J . Mahler, Duarte R. Ziegler, Ulm

306 Figures in 327 Single figures

Volume 6/1974 and210Tables

Georg Thieme Verlag Stuttgart

III

TABLE OF CONTENTS

M e l a n i e , F . Prohormones in Tissues and in Circulation . . . 1 Miyai, K., M. Azukizawa and Y. K u m a h a r a Bioassay and Immunoassay Measurements of Serum Thyrotropin in Primary Hypothythyroid-ism Before and After the Administration of Thyrotropin Releasing Hormone 9

Moody, A . J . , M . A . Stanlf, M. Stan, J. G l i e m a n n A Simple Free Fat Cell Bioassay for Insulin . . . 12

Serrano-Rios, M., F G . H a w k i n s , F. Escobar, J.M. M a t o , L . L a r r o d e r a , M. de O y a and J . L . R o d r i g u e z - M i n o n Insulin Secretion in Addison's Disease: Effect of Hydrocortisone Treatment 17

Lücke, C., B . Höffken, H J . M i t z k a t and I. Trau tschold Effect of 2-Deoxy-D-Glucose and Mannoheptu-lose on the Insulin Response to Pancreozymin in Rabbits 22 Lisch, H.-J., S. Sailer, F. Sandhofen R . Tschikof and H. Braunsteiner The Action of Insulin and Glucose on Lipolysis in Isolated Human Fat Cells 25 Werner, S. and H Low Inhibitory Effects of Calcitonin on Lipolysis and ^Calcium Accumulation in Rat Adipose Tissue in Vivo 30

Schleyer, M., K L . F e h m , K . H Voigt, J.D. Faul­haber and E.F. Pfeiffer Studies on the Pituitary "Fettstoffwechselhor­mon" 36

Das, I. and D . A . Hems Glycogen Synthetase and Phosphorylase Activ-ities in Different Tissues of Genetically Obese Mice 40

Goebell, H The Effect of Ethionine on the Energy-Produc-ing Metabolism in the Rat Pancreas 44

H o w e l l , S . L . , J.C. Edwards and W. M o n t a g u e Regulation of Adenylate Cyclase and Cyclic-AMP Dependent Protein Kinase Activities in A2-Cell Rieh Guinea-Pig Islets of Langerhans . 49

R o l d a n , A . G . , E J . del Castillo and V.G. Foglia Effect of Cyclic AMP on Ketogenesis in Liver Homogenates from Pancreatectomized-Hypo-physectomized Dogs 53

R o l l e r i , E . , R . M a l v a n o , C. Gandolfi and U. Rosa Effect of Irreversible Adsorbents on the Disso-ciation of the Antigen-Antibody Complexes . 57

GunZaga, J., Th. Lauffenburger, C h a r l o t t e Lent-ner, M . A . Dambacher, H.G. Haas and H F l e i s c h , A . J . Olah Diphosphonate Treatment of Paget's Disease of Bone 62 Gautvik, K.M. and A . H . Tashjian, Jr. Human Medullary Thyroid Carcinoma: Control of Calcitonin Secretion In Vivo and in Tissue Culture 70

Boonayathap, U. and S.F. M a r o t t a Renal Handling of Cortisol in Dogs 74

Gennser, G., P. L i e d h o l m and J. T h o r e i l Pituitary Responses to Continuous Administra­tion of LRH in Human Males with Oligozoo-spermia 79

Eshet, R . , Z. L a r o n , M. B r o w n and R . A r n o n Immunological Behaviour of HGH from Plasma of Patients with Familial Dwarfism and High IR-HGH in a Radioimmunoassay System Using the Cross-Reaction between HGH and HCS . . . 80

K e r p e l - F r o n i u s , E . , G. Gäcs and Ch. H e r v e l Pituitary Function in Malnutrition 82

Legros, J.J. and P. Lefebvre Lack of Effect of Human Neurophysin I on the Metabolism of Rat Adipose Tissue 83

Renauld, A . , R . C . Sverdlik and L . L . Andrade Effect of Thyroidectomy upon the Insulin Tol-erance Test 84

Bajaj, J.S. and J. Vallance-Owen Fenfluramine and Glucose Uptake by Muscle . 85

F r a y n , K . N . , A n m a r i e Hedges and Marilyn J. Kirby Stimulation by Fenfluramine of Glucose Uptake into Skeletal Muscle In Vitro 86

Bercovitch, L and J . A . Moorhouse Effect of Posture on Plasma Cortisol Level . . . 87

R a h e j a , K . L . and E.F. Reber Relative Ineffectiveness of Exogenous as Com-pared with Endogenous Testosterone in Combat-ing Hypercholesterolemia 88

Sutherland, D.J.B, and R . L . S i n g h a i Stimulation of Prostatic Adenyl Cyclase by Di­hydrotestosterone 89

L e r n e , C.E., B . L . Wajchenberg, M . O . R . Leite and A . B o r e l l i The Dissociation of Urinary Sodium and Cal­cium Excretions 90 Rayssiguier, Y. and P. L a r v o r Parathyroid Response to Hypocalcemia in Mag­nesium Deficient Rat 91

IV

S u l m a n , G. and E l i Tal Treatment of Functional Hypothyroidism by Oral TRF Monitored by Daily Urinary Thyrox­ine and Histamine Assay 92

G h i o n e , S.t M. Pellegrini, G. Buzzigoli, A . Carpi, C. Valori and L . D o n a t o Plasma and Urinary Catecholamine Levels and Thyroid Activity in Relation to Cardiovascular Changes in Hyper- and Hypothyroids 93

de Reviers, M. and M.P. Dubois Binding of Synthetic-LHRF Antibodies in the Median Eminence of the Cockerel 94

K a l k , W.J., A . I . Vinik, S. B a n k , P. K e l l e r a n d W.P. U. Jackson Selective Loss of Beta Cell Response to Glu­cose in Chronic Pancreatitis 95

Bruckdorfer, K . R . , S.S. Kang, L H K h a n , A . R . B o u r n e and J. Y u d k i n Diurnal Changes in the Concentrations of Plas­ma Lipids, Insulin and Corticosterone in Rats fed Diets Containing Various Carbohydrates . 99 Tjälve, H , D . Popov and P. Slanina Effect of the Local Anesthetics Procaine and Lidocaine on Insulin Secretion from Rabbit Pancreas Pieces 106

Loubatieres, A . L . , M M . L o u b a t i e r e s - M a r i a n i , R . A l r i c , G. Ribes and H Agot Studies of the Action of Neutral Red on Glyc-emia and on Insulin and Glucacon Secretion in the Dog 111

Aynsley-Green, A . and K G . M M . A l b e r t i In Vivo Stimulation of Insulin Secretion by Guanidine Derivatives in the Rat 115

H o w l a n d , B . E . and E J . Zebrowski Serum and Pituitary Gonadotropin Levels in Alloxan-Diabetic Rats 121

Roos, P , J M . M a r t i n , S. Westman-Naeser and C. Hellerström Immunoreactive Growth Hormone Levels in Mice with the Obese-Hyperglycemic Syndrome (Genotype obob) 125 R a b k i n , R . , Margaret S w a n n , D J . Shapiro and L . Isaacson Effect of Growth Hormone on Sodium Trans­port and Osmotic Water Flow AcrossToadSkin . 129

Pokroy, N . , S. Epstein, Salle H e n d r i c k s and B . Pimstone Thyrotrophin Response to Intravenous Thyro-trophin-Releasing Hormone in Patients with Hepatic and Renal Disease 132

Renauld, A . , R . C . Sverdlik a n d L . L . Andrade Effects of Hypothyroidism on Serum Immuno­reactive Insulin, Free Fatty Acids and Blood Sugar in the Dog as Tested for Oral Glucose Tolerance. Corrective Effects to Thyroxine Therapy 137

Adkofer, F., U. Armbrecht and H Schleusener, with technical assistance f r o m C. Brammeier and M M . Schürnbrand Plasma Lecithin: Cholesterol Acyltransferase Activity in Hypo- and Hyperthyroidism . . . . 142

G o r i n , E . and H M . Goodman Protein Kinase in Adipose Tissue: Effect of Hypophysectomy 146

Elkeles, R . S . The Role of Heparin in Lipoprotein Lipase Activity 151

Boonayathap, U. and S.F. M a r o t t a Renal Handling of Cortisol in Dogs. II. Effect of ACTH on Stop Flow Pattern 154

Siegmund, P , A . Tüllmann and M. H o l k e with the technical collaboration of P. Ott and L . Uher The Cyclic AMP-Mediated Action of Epinephri-ne and the Activity of Carbonic Anhydrase in Avian Erythrocytes 158

O f D o r , R . K . , F E . Newsome, C O . Parkes and D.H. Copp Peptides Obtained from Porcine Calcitonin After Degradation by a Human Plasma Frac-tion 161 Berger, W., M. Stahl, E. O h n h a u s and H Gaschke Pancreatic Glucagon, Plasma Insulin and Blood Glucose Response to Arginine Infusion in Non-diabetic Subjects Following Biguanide Pretreat-ment 165

Davis, B . and N . R . Lazarus Effect of Glucagon, Diazoxide, Phentolamine on Islet Adenylate Cyclase 166

Strohfeldt, P , H Kettl and K.F. Weinges Perfusion of the Isolated Rat Hindlimb with a Synthetic Medium 167

M e i n h o l d , H. and K. W. Wenzel Radioimmunoassay of Thyroxine in Unextract-ed Serum 169

Nieschlag, E . , T Walk and A . E . Schindler Dehydroepiandrosterone (DHA) and DHA-Sulfate during Pregnancy in Maternal Blood. . 170

Mauss, J. and G. Borsch Clomiphene Citrate Stimulation in Male Sub-and Infertility Monitored by Simple Immuno-chemical Assays for FSH and LH in Uncon-centrated Urine 171

N e r u p , J., O. Ortved Andersen, G. Bendixen, J. Egeberg, R . Gunnarsson, H. K r o m a n n and J.E. Poulsen Glucose Intolerance and Islet Damage in Mice Immunized with Homologous Endocrine Pancreas — A Preliminary Communication . . 173

V

K u m a r , D . , S . D . M e h t a l i a and L . V. Miller Antigenicity of Monocomponent Porcine Insulin in Rabbits 175

Pento, J.T., A . Kagan and S M G l i c h Influence of Altered States of Calcium Homeo-stasis on Insulin Secretion in Rats and Rabbits 177

Somogyi, J., I. Vincze, F. Willig and F . K Schmidt The Subcellular Distribution of ^-Gliben­clamide in Different Tissues of Rat 181 Tchobroutsky, G., M.-E. Lenormand, G. M i c h e l and R . Assan Lack of Post Prandial Exercise-Induced Growth Hormone Secretion in Normoglycemic Insulin-Treated Diabetic Men 184

Clemens, J . A . , C J . Shaar, E . B . Smalstig and C. Matsumoto Effects of Some Psychoactive Agents on Pro-lactin Secretion in Rats of Different Endocri-ne States 187

V H e r m i t e , M . , C Robyn, J. Golstein, G. Rothenbuchner, J. B i r k , U. Loos, M. Bonnyns and L . Vanhaelst Prolactin and Thyrotropin in Thyroid Diseases: Lack of Evidence for a Physiological Role of Thyrotropin-Releasing Hormone in the Regula­tion of Prolactin Secretion 190

P a v l o v i c - H o u r n a c , M. and D . Delbauffe In Vivo Incorporation of Labelled Amino Acid into Different Protein Fractions of the Rat Thyroid Gland 196

Busnardo, B . , M.E. Girelli, L . V a r o t t o and A . Giacobbi Computer Analysis of Early Thyroxine Distri­bution in Thyrotoxicosis and Hypothyroidism 202 Klenerova, V. and S. Kynie The Activity of Adenylate Cyclase, Phospho­diesterase and Protein Kinase in the Adeno-hypophyses of Rats Treated by Chronic Ad­ministration of Estrogenes and Acute Appli­cation of Thyroxine 208

Kley, KK, E. Nieschlag, F. B i d l i n g m a i e r and K L . Krüskemper Possible Age-Dependent Influence of Estro­gens on the Binding of Testosterone in Plasma of Adult Men 213

Sladek, C D . Gluconeogenesis and Hepatic Glycogen For­mation in Relation to the Rat Estrous Cycle . 217

Adolfsson, S., S. Boström, M. F a h l e n , A . Hjalmarson, B . E . Hustvedt and R . Johansson Enzyme Activities in Muscle and Liver After Destruction of the Ventromedial Hypothala­mus Area and Administration of Insulin 222

M a c h o , L . The Response of Adipose Tissue to Lipid Mobilizing Hormones in Postnatally Under-and Overnourished Rats 226

Gilbert, C . K and D J . G a l t o n The Effect of Catecholamines and Fasting on Cyclic-AMP and Release of Glycerol from Human Adipose Tissue 229 Cox, B . D . , A . R . Clarkson, M J . Whichelow and P. R u t l a n d Effect of Adrenaline on Plasma Vitamin C Levels in Normal Subjects 234

Ward, W.F., K.M. K l i t g a a r d and R . C Meade Insulin Degradation by Fat Cell Ghosts . . . . 238

H e r r m a n n , J, H J . Rusche, H J . Kröll, P. Hä­ger and K L . Krüskemper Free Triiodothyronine (T3)- and Thyroxine (T4)-Serum Levels in Old Age 239

Galloway, D . G . and J. Pelletier Influence of Age on the Pituitary Response of Male Lambs to Synthetic LH-RH Injection . . 240

Wickramasinghe, R . K Effect of 11-Deoxycorticosterone on Choles-terol Sidechain Cleavage by Adrenal Cortex Mitochondria 241

Jerums, G. and D J . G a l t o n Two Types of Insulin Receptors Mediating Opposite Effects on Cyclic-AMP in Fat Cells of the Rat 242 G o z a r i u , L . , K. Forster, J . D . F a u l h a b e r , H. M i n n e and R . Ziegler Parathyroid Hormone and Calcitonin: Influ-ences upon Lipolysis of Human Adipose Tissue 243

T i e n g o , A . , D . Fedele, P. Frasson, M. Muggeo and G. Crepaldi Ethanol Effect on Glucagon Secretion in the Pig 245

Kissebah, A . K , N. Vydelingum, B . R . T u l l o c h , H H o p e - G i l l and T R . Fräser The Role of Calcium in Insulin Action. I. Purification and properties of enzymes regulat-ing lipolysis in human adipose tissue: effects of cyclic-AMP and calcium ions 247

G o b e r n a , R . , J. T a m a r i t Jr., J. Osorio, R . Fuss­gänger, J. T a m a r i t and E . F . Pfeiffer Action of B-Hydroxy Butyrate, Acetoacetate and Palmitate on the Insulin Release in the Perfused Isolated Rat Pancreas 256

Rehfeld, J.F. and J. Iversen Secretion of Immunoreactive Gastrin from the Isolated, Perfused Canine Pancreas . . . . 260

VI

Alemany, M. and E . Herr era Effects of Suckling and Food Deprivation on the Acitivity of Citrate Enzymes in the Liver of the Rat 264

H o w e l l , S . L . , C. Hellerström and M. Tyhurst Intracellular Transport and Storage of Newly Synthesised Proteins in the Guinea Pig Pan-creatic A Cell 267

Z e r m a t t e n , A . and J.-P. F e l b e r Sensitivity to Glucose of an Intestinal Factor Stimulating Insulin Release 272

Vargas, L . , M. Bronfman and M.E. K a w a d a Stress, Insulin Antagonist and Transient Dia­betes Mellitus in the Rat 275

Tarachand, U. and J. E a p e n Effect of Streptozotocin-Diabetes on In Vivo Protein Synthesis by Placenta and Liver of Mice 280

Schönborn, J., W. Poserf U. Panten and A . Hasselblatt Effect of Hypoglycemic Sulfonylureas on Hepatic Fructose Metabolism in the Rat . . 284

Rado, J.P.y L . Borbely, L . Szendet J. Fischer and J. T a k o Investigation of the Diuretic Effect of Gliben­clamide in Healthy Subjects and in Patients with Pituitary and Nephrogenic Diabetes Insipidus 289

M a r s c h n e r , L , P. B o t t e r m a n n , F. E r h a r d t , R . L i n k e , G. Löffler, V. Maier, P. Schwandt, W. Vogt and P C . Scriba Group Experiments on the Radioimmunolo-gical Insulin Determination 293 S u t t e r - D u b , M.Th.f R . Leclercq, B . Ch.J. Sutter and R . Jacquot Plasma Glucose, Progesterone and Immuno-reactive Insulin Levels in the Lactating Rat . 297

Hervas, F. and G. M o r r e a l e de Escobar A Rapid Procedure for the Radioimmuno-assay of Rat Growth Hormone 300

Kowadlo Silbergeld, A . and Z. L a r o n Lipolytic Effects of Sheep (3-Lipotropin in Rat Adipose Tissue: Interaction with Theophylline and Dihydroergotamine 303

Berthezene, F., B . Chavrier and R . M o r n e x Increase in the Amount of Iodide Released from the Thyroid after Perchlorate or Car­bimazole Administration 306

Jortay, A . M . , F . R . L . C a n t r a i n e and J.E. D u -mont Iodide Trapping by Thyroid Slices in Vitro . . 309

Lilienfeld-Toal, H v., R . - D . Hesch, M. Hüfner and Chr. Mclntosh Excretion of Cyclic 3', 5'-Adenosine Monophos-phate in Renal Insufflciency and Primary Hyper-parathyroidism after Stimulation with Para-thyroid Hormone 314

Werner, S., K. Hall and H Low Similar Effects of Calcitonin, Insulin and So-matomedin A on Lipolysis and Uptake of Cal­cium and Glucose in Rat Adipose Tissue In Vitro 319

Gleispach, H. The Urinary Steroid Excretion in Girls during Puberty 325

M a r o t t a , S . F . , D . M . L a n u z a and L . G . Hiles Diurnal Variations in Plasma Corticosterone and Cations of Male Rats on Two Lighting Schedules 329

Bates, R . F . L . and J.P. B a r l e t The Preventive Effect of Porcine Calcitonin given by Mouth on Restraint-Induced Gastric Ulcer in Rats 332

Kappus, H and H M . Bolt Irreversible Binding of Ethynylestradiol Meta­bolites to Protein: Lack of Methylation by S-adenosyl-Methionine 333

Kato, H , T I t o , Y. Kido and T Torigoe Preparation of 1251-Labeled Estrone-Tri Ty-rosine Methyl Ester 334

Rastogi, G.K., J. C h a k r a b o r t i and M.K. S i n h a Serum Gonadotropins (LH and FSH) and their Response to Synthetic LHRH in Diabetic Men with and without Impotence 335

Mohnfeld, G., J. Mauss and G. Borsch Synthetic LH-RH Stimulation Monitored by Immunochemical Assay for Urinary FSH and LH 336

Voigt, K . H . , H G . D a h l e n , H L . F e h m , J. B i r k , K. Schröder, H P . G . Schneider, G. Rothen-buchner and E.F. Pfeiffer Simultaneous Stimulation Test for the Anterior Pituitary Hormones 337

Pfeiffer, E . F , Ch. T h u m and A . H Clemens The Artificial Beta Cell 339

I m u r a , H., Y. N a k a i , Y. Kato, Y. Yoshimoto and K. M o r i d e r a Propranolol-Insulin Stimulation Test in the Diagnosis of Growth Hormone Deficiency . . 343

B e r l e , P . , E. Finsterwalder and M. Apostolakis Comparative Studies on the Effect of Human Growth Hormone, Human Prolactin and Human Placental Lactogen on Lipid Metabo­lism 347

VII

Ogundipe, 0.0. and A . Bray The Influence of Diet and Fat Cell Size on Glucose Metabolism, Lipogenesis and Lipolys­is in the Rat Kissebah, A . H . , B . R . T u l l o c h , N . Vydelingum, H Hope-Gill, P. Clarke and T . R . Fräser The Role of Calcium in Insulin Action . . . . Werner, S, and H Low Adenylate Cyclase in Rat Adipocyte Plasma Membranes after Adrenalectomy and Adminis­tration of Cortisone Acetate

G o l d m a n , J.K. and L . L . Bernardis Metabolism of Glucose, Fructose and Pyruvate in Tissues of Weanling Rats with Hypothalamus Obesity

F r a n c o i s e R a b a i n and L . Picon Effect of Insulin on the Materno-Fetal Trans­fer of Glucose in the Rat

A n a A r a n d a and E. Herr era The Effect of Food Deprivation on In Vivo Gluconeogenesis in the Suckling Rat

Gaschke, H , A . Denes, J. Girard, F. Collard and W. Berger Circadian Variations of Carbohydrate Toleran-ce in Maturity Onset Diabetics Treated with Sulfonylureas

Massi-Benedetti, F., A . Falorni, A . Luyckx and P. Lefebvre Inhibition of Glucagon Secretion in the Hu­man Newborn by Simultaneous Administra­tion of Glucose and Insulin Vinik, A . L and A . Hardcastle Structure Antigenicity Relationships of Gluca­gon and Related Peptides

Renauld, A . , L . L . Andrade, R . C . Sverdlik and R . R . Rodriguez Serum Insulin Response to Glucose Infusion in Hyperthyroid Dogs

Story, J . A . and D . R . Griffith Effect of Exercise on Thyroid Hormone Secretion Rate in Aging Rats

Becker, F . O . , F. B u c h a n a n and M. Rasho Parathyroid Hormone Degradation Studies in Maternal and Fetal Rat Tissues by Radioimmu-noassay

Leicht, E , G. B i r o and K.F. Weinges Inhibition of Releasing-Hormone-Induced Secretion of TSH and LH by Calcitonin . . . .

B o r r e i l , J and S. B o r r e l l Alterations of Adrenal Corticosteroids and Catecholamines Düring Arnino-Glutethimide Feeding

S u v a r n a l a t h a , M. and H.B. Devaraj S a r k a r Effect of Steroids on Hemicastration In-duced Compensatory Hypertrophy of the

351 Testis in the Frog, Rana hexadactyla (Lesson) 417

Debry, G., P. D r o u i n and L . Mejean 357 Influence of Triglyceridemia Level on Glucose

Tolerance and Insulinemia in Subjects with Type IV Hyperlipoproteinemia 421

Bittner, R . , H G . Beger and E . Kraas 365 Osciilation in Insulin Response to Glucose

Stimulation 423

Sundby, F . , J. Markussen and W. D a n h o Camel Glucagon: Isolation, Crystallization

370 and Amino Acid Composition 425

L a u b e , H , R . D . Fußgänger and E.F. Pfeiffer Paradoxical Glucagon Release in Obese Hyper-

376 glycemic Mice 426

Velasco, C, W. O p p e r m a n n , N. Marine and R . Camerini-Davalos

381 Effect of Genetic Diabetes on Kidney Gluco-syltransferase 427

Sutton, J. and L . Lazarus Effect of Adrenergic Blocking Agents on Growth Hormone Responses to Physical Exercise . . 428

3 8 6 Rado, J.P., L . Vegh and I r e n e Sawinsky Interference of Triamterene with Cortisol Determination in Urine 429

Yang, M.M.P. and S . H Kok Effect of a Single Dose of Progesterone on Blood Glucose in Diabetic Rats and on Insulin Sensitivity in Normal Rats 430

Bolt, W.H, H. Kappus and H M . Bolt Ring A Oxidation of 17a-Ethynylestradiol in Man 432

Steiner, H , M. Z a n i s i and L . M a r t i n i Antiovulatory Activity of the Thyrotropin Re-leasing Hormone in the Rat 432

Cabello, G. and M.C. M i c h e l Plasmatic Hormonal Iodine in Healthy and

4 0 3 Diarrheic Calves 434

Sawhney, R . C . and G.K. Rastogi Estimation of TBG by a Competitive Ligand-Binding Assay 435

Voigt, K.H., H G . D a h l e n , H L . F e h m , L . B i r k , K. Schröder, H P . G . Schneider, G. Rothenbuch-ner and E.F. Pfeiffer Simultaneous Stimulation Test for the Anterior Pituitary Hormones 436

F i n k , G., J.C. Cresto, R . A . G u t m a n , R . I . L a v i n e , A . H . Rubenstein and L . Recani Plasma Proinsulin-Like Material in Insulin Treat­ed Diabetics 439

VIII

R a n c o n , F . , M. L a b u r t h e , G. Rosselin and P. Freychet Untractable Hypoglycemia in an Infant: Studies on Pancreas Insulin and Glucagon 443 A n d r e a n i , D . , M. I a v i c o l i , G. T a m b u r r a n o and G. M e n z i n g e r Comparative Trials with Monocomponent (MC) and Monospecies (MS) Pork Insulins in the Treatment of Diabetes mellitus. Influence on Antibody Levels, on Insulin Requirement and on Some Complications 447

G o l d m a n t J.K. and L . L . Bernardis Persistent Adipose Tissue Insulin Responsive-ness Düring Fasting of Weanling Rats with Hypothalamus Obesity 454

H o p e - G i l l H , N. Vydelingum, A . H Kissebah, B . R . T u l l o c h and T R . Fräser Simulation and Enhancement of the Adipose Tissue Insulin Response by Procaine Hydro-chloride: Evidence for a Role of Calcium in Insulin Action 457

Rosak, C., E. Haupt, J. Beyer and K. Schaff­ung Investigations on the Antilipolytic Activity of Sulfonylureas in Man with Indications of Limit Dosages Concerning their Insulin-Secret-ing Properties 464

Lambert, A . E . , J.C. H e n q u i n and P. Malvaux Cationic Environment and Dynamics of Insulin Secretion. IV. Effect of Ouabain 470

Schimmel, R . J . and D . G r a h a m Inhibition by Diphenylhydantion of the Dia-betogenic Action of Streptozotocin 475

O h n e d a , A . , M. Sato, K. M a t s u d a , H I t a b a s h i , K. H o r i g o m e , M. C h i b a and S. Yamagata Suppression of Pancreatic Glucagon Secretion by Tolbutamide in Dogs 478

K r a l , J . G . : The Effects of Different Methods of Sacrifice on Lipid and Carbohydrate Meta­bolism in the Rat 483

Boninsegna, A . , G. Federspil and C. D e P a l o The Effect of Muscular Exercise on Free Fatty Acids, Acetoacetate and 3-Hydroxy-butyrate Blood Levels 488

Tarachand, U. and J. E a p e n A Comparative Study on In Vivo Protein Syn-thesis by Liver of Pregnant and Non-pregnant Mice 491

Schimpff, R . M . , M. D o n n a d i e u , M. Gourmelen and F. Girard The Effects of hGH Treatment on Somatome-din Levels in the Serum (as determined by 3 5 S uptake into cartilage) 494

Vinik, A . L , W.J. K a l k , H M c L a r e n and M. Paul Impaired Prolactin Response to Synthetic Thyro-tropin-Releasing Hormone after a 36 Hour Fast . 499

Rothenbuchner, G., D A . Koutras, S. Raptis, J. B i r k , U. Loos, G. Rigopoulos and B. Mala-mos The Effect of Thyrotrophin-Releasing Hormone on Serum TSH, T 4 , and T 3 Levels in Endemie and Sporadic Nontoxic Goitre 501 Golstein, J., E . Van Cauter, R . Leclercq, G. Copinschi and L . Vanhaelst Monthly Variations of Serum Levels of Thyro­tropin, Growth Hormone in Cortisol in Man . 506

D a h l e n , H G . , E. Keller and H.P.G. Schneider Linear Dose Dependent LH Release Following Intranasally Sprayed LRH 510

Baier, H , B i r o and K.F. Wernges Serum Levels of FSH, LH and Testosterone in Human Males 514 B a r l e t , J.P.: The Influence of Porcine Calci­tonin Given Intragastrally on Restraint-In-duced Gastric Ulcers in Pigs . 517

A n t o n i o , P . , M. G a b a l d o n , T Lacomba and A . Juan Effect of the Antiestrogen Navoxidine on the Occurrence of Estrogen-Dependent Renal Tu­mors in Hamster 522

Clarke, P , A . Kissebah, N. Vydelingum, H Hope-Gill, B . T u l l o c h and R . Fräser Regulation of Glycogenolysis through Changes in Intracellular Calcium 525

Vinik, A . L and S. Jessop Red Blood Cell Potassium and Insulin Release 526

Dolais-Kitabgi, Jacqueline, M. L a b u r t h e and G. Rosselin Immunological Specificity of the ß Subunit of the Human Chorionic Gonadotropin (HCG) 527

Rastogi, G.K., R . C . Sawhney, N.C. P a n d a and B. B. Tripathy Thyroid Hormone Levels in Adult Protein Ca-lorie Malnutrition (PCM) 528

Thomas, J . A . and M. Manandhar Effect of Prolactin and/or Testosterone on Cyclic AMP in the Rat Prostate Gland . . . . 529

y ' / Rado, J.P., T Simon, E. Juhos, J. T a k o and O. Nagy Interference of Psychotropic Drugs with Cortisol Determinations 530 Sönka, J., J. Kopeckä, A . Pavlovd, A . Z b i r k o -vä and J. Stas Effects of Diet and Exercise on Catecholamine Excretion 532 Meyer, Jr., R . A . and Martha H Meyer Increased Liver Inorganic Phosphate Following Thyrocalcitonin Administration to Rats . . . . 533

Offermann, G. and F. D i t t m a r A Direct Protein-Binding Assay for 25-Hydro-xycalciferol 534

IX

AUTHOR'S INDEX

A Adlkofer, F. 142 Adolfsson, S. 222 Agot, H. 111 Alberti, K.G.M.M. 115 Alemany, M. 264 Alric, R. 111 Andrade, L .L . 84, 137, 400 Andreani, D. 447 Antonio, P. 522 Apostolakis, M. 347 Aranda, ANA. 381 Armbrecht, U. 142 Arnon, R. 80 Assan, R. 184 Aynsley-Green., A . 115 Azukizawa, M. 9

B Baier, H. 514 Bajaj, J.S. 85 Bank, S. 95 Barlet, J.P. 332, 517 Bates, R.F.L. 332 Becker, F.O. 407 Beger, H.G. 423 Bendixen, G. 173 Bercovitch, L. 87 Berger, W. 165, 386 Berle, P. 347 Bernardis, L .L. 370, 454 Berthezene, F. 306 Beyer, J. 464 Bidlingmaier, F. 213 Birk, L 190, 337,436,501 Biro, G. 410, 514 Bittner, R. 423 Borsch, G. 171, 336 Bolt, H.M. 333 Boninsegna, A . 488 Bonnyns, M. 190 Boonayathap, U. 74, 154 Borbely, L. 289 Borreil, J. 414 Borreil, S. 414 Borelli, A. 90 Boström, S. 222 Bottermann, P. 293 Bourne, A.R. 99 Braunsteiner, H. 25 Bray, A. 351 Bronfman, M. 275 Brown, M. 80 Bruckdorfer, K.R. 99 Buchanan, F. 407 Busnardo, B. 202 Buzzigoli, G. 93

C Cabello, G. 434 Camerini-Davalos, R.A. 427 Cantraine, F.R.L. 309 Carpi, A. 93 Castülo, Del. E J . 53 Cauter, E. van 506 Charkaborti, J. 335 Chavier, Bernadette 306 Chiba, M. 478 Qakson, A.R. 234

Clarke, P. 357, 525 Clemens, A.H. 339 Clemens, J.A. 187 Collard, F. 386 Copinschi, G. 506 Copp, D.H. 161 Cox, B.D. 234 Crepaldi, G. 245 Cresto, J.C. 439

D Dahlen, H.G. 436, 510 Dahlen, H.L. 337 Dambacher, M.A. 62 Danho, W. 425 Das, I. 40 Davis, B. 166 Debry, G. 421 Delbauffe, Daniele 196 Denes, A . 386 De Pala, C. 488 Devaraj-Sarkar, H.B. 417 Dittmar, F. 534 Dolais-Kitabgi, Jacqueline 527 Donato, L. 93 Donnadieu, M. 494 Drouin, P. 421 Dubois, M.P. 94 Dumot, J.E. 309

E Eapen, J. 280, 491 Edwards, J.C. 49 Egeberg, J. 173 Elkeles, R.S. 151 Epstein, S. 132 Erhardt, F. 293 Escobar, F. 17 Eshet, R. 80

F Fahlen, M. 222 Falorni, A. 392 Faulhaber, J.D. 36/243 Fedele, D. 245 Federspil, G. 488 Fehm, H.L. 36, 337/436 Felber, J.R. 272 Fink, G. 439 Finsterwalder, E. 347 Fischer, J. 289 Fleisch, H. 62 Foglia, V.G. 53 Forster, K. 243 Fräser, R. 525 Fräser, T.R. 247, 357, 457 Frasson, P. 245 Frayn, K .N . 86 Freychet, P. 443 Fussgänger, R. 256 Fussgänger, R.D. 426

G Gabaldon, M. 522 Gäcs, G. 82 Galloway, D.G. 240 Galton, D.J. 229, 242 Gandolfi, C. 57 Gautvik, K .M. 70

Gennser, G. 79 Ghione, S. 93 Giacobbi, A . 202 Gilbert, C.H. 229 Girard, F., 494 Girard, J., 386 Girelli, M.E. 202 Gleispach, H., 325 Glick, S.M. 177 Goberna, R. 256 Goebell, H. 44 Göschke, H. 386 Goldman, J.K. 370, 454 Golstein, J. 190, 506 Goodman, H.M. 146 Gorin, E. 146 Goschke, H. 165 Gourmelen, M. 494 Gozariu, L. 243 Graham, D. 475 Griffith, D.R. 403 Guncaga, J. 62 Gunnarsson, R. 173 Gutman, R.A. 439

H Haas, H.G. 62 Hau, K. 319 Hardcastle, A. 396 Hasselblatt, A. 284 Haupt, E. 464 Hawkins, F.G. 17 Hedges, Annmarie 86 Hellerström, C. 125, 267 Hems, D.A. 40 Hendricks, Salie 132 Henquin, J.C. 470 Hermite, M.L. 190 Herrera, E. 264, 381 Hervas, F. 300 Hervei, Ch. 82 Hesch, R.D. 314 Hialmarson, A. 222 Hiles, L.G. 329 Höffken, B. 21 Holke, M. 158 Hope-Gill, H. 247, 357, 457, 525 Horigome, K. 478 Howell, S.L. 49, 267 Howland, B.E. 121 Hüfner, M. 314 Hustvedt, B.E. 222 Hynie, S. 208

I Iavicoli, M. 447 Imura, H. 343 Isaacson, L. 129 Itabashi, H. 478 Ito, T. 334 Wersen, J. 260

J Jackson, W.P.U. 95 Jacquot, R. 297 Jerums, G. 242 Jessop, S. 526 Johansson, R. 222 Jortay, A .M. 309

X

Juan, A. 522 Juhos, E. 530

K

Kagan, A . 177 Kalk, W.J. 95, 499 Kang, S.S. 99 Kato, H. 334 Kato, Y . 343 Kappus, H. 333 Kawada, M.E. 275 Keller, E. 510 Keller, P. 95 Kerpel-Fronius, E. 82 Kettl, H. 167 Khan, L H . 99 Kido, Y . 334 Kirby, Marilyn, J. 86 Kissebah, A. 525 Kissebah, A . H . 247, 357, 457 Klenerova, V. 208 Kley, H .K. 213 Klitgaard, H .M. 238 Kok, S.H. 430 Kopecka, J. 532 Koutras, D.A. 501 Kowadlo Silbergeld, A . 303 Kraas, E. 423 Kral, J.G. 483 Kromann, H. 173 Krüskemper, H .L. 213 Kumahara, Y. 9 Kumar, D. 175

L Laburthe, M. 443, 527 Lacomba, T. 522 Lambert, A.E . 470 Lanuza, D.M. 329 Laren, H. , Mc. 499 Laron, Z. 80, 303 Larrodera, L. 17 Larvor, P. 91 Laube, H. 426 Lauffenburger, Th. 62 Lawine, R.L. 439 Lazarus, L. 428 Lazarus, N.R. 166 Leclercq, R. 297, 506 Lefebvre, P. 83/392 Legros, J.J. 83 Leicht, E. 410 Leite, M.O.R. 90 Lerne, C E . 90 Lenormand, Marie-Emmanuelle 184 Lentner, Charlotte 62 Liedholm, P. 79 v. Lilienfeld-Toal, H. 314 Linke, R. 293 Lisch, H.-J. 25 Löffler, G. 293 Low, H. 30/319/365 Loos, U . 190, 501 Loubatieres, A . L . 111 Loubatieres-Mariani, M.M. 111 Lücke, C. 21 Luyckx, A. 392

M Macho, L. 226

Maier, V. 293 Malmos, B. 501 Maltarello, C , Miss 447 Malvano, R. 57 Malvaux, P. 470 Manandhar, M. 529 Marine, N . 427 Markussen, J. 425 Marotta, S.F. 74, 154, 329 Marschner, I. 293 Martin, J.M. 125 Martini, L. 432 Massi-Benedetti, F. 392 Mato, J .M. 17 Matsuda, K. 478 Matsumoto, C. 187 Mauss, J . 171, 336 Mclntosh, Chr. 314 Meade, R.C. 238 Mehtalia, S.D. 175 Meinhold, H. 169 Mejean, L. 421 Melani, F. 1 Menzinger, G. 447 Meyer, Martha, H. 533 Meyer, Ralph, A . Jr. 533 Miayai, M. 9 Michel, G. 184 Michel, M.C. 434 Miller, L .V . 175 Minne, H. 243 Mitzkat, H J . 21 Mohnfeld, J. 336 Montague, W. 49 Moody, A.J . 12 Moorhouse, J.A. 87 Moridera, K. 343 Mornex, R. 306 Morreale, de Escobar, G. 300 Muggeo, M. 245

N Nagy, O. 530 Nakai, Y . 343 Nerup, J. 173 Newsome, F.E. 161 Nieschlag, E . 170, 213

O O'dor, R.K. 161 Off ermann, G. 534 Ogunipe, O.O. 351 Ohneda, A . 478 Ohnhaus, E. 165 Olah, A.J . 62 Oppermann, W. 427 Ortvedandersen, O. 173 Osorio, J. 256 Oya, de, M. 17

P Panda, N.C. 528 Panten, U. 284 Parkes; C O . 161 Paul, M. 499 Paulsen, J.E. 173 Pavlova, A . 532 Pavlovig, Mira. 196 Peletier, J . 240 Pellegrini, M. 93 Pento, J.T. 177

Pfeiffer, E.F. 36, 256, 337, 339, 426,436 Picon, L. 376 Pimstone, B. 132 Pokroy, N . 132 Popov, D. 106 Poser, W. 284

R Rabain, Francoise 376 Rabkin, R. 129 Rado, J.P. 289,429,530 Raheja, K . L . 88 Rancon, F. 443 Raptis, S. 501 Rasho, M. 407 Rastogi, G.K. 335,435,528 Rayssiguier, Y. 91 Reber, E.F. 88 Recant, L. 439 Rehfeld, J.F. 260 Renauld, A . 84, 137,400 Reviers, de. M. 94 Ribes, G. 111 Rigopoulos, G. 501 Robin, C. 190^ Rodriguez-Minon, J.L. 17 Rodriguez, R.R. 400 Roldan, A . G . 53 Rolleri, E. 57 Rosa, U. 57 Rosak, C 464 Roos, P. 125 Rosselin, G. 443,527 Rothenbuchner, G. 190, 337,436, 501 Rubenstein, A . H . 439 Rutland, P. 234

S Sauer, S. 25 Sandhofer, F. 25 Sato, M. 478 Sawhney, R.C. 435,528 Sawinsky, I. 429 Schimpff, R .M. 494 Schimmel, R.J. 475 Schindler, A .E . 170 Schleusener, H. 192 Schleyer, M. 36 Schmidt, F .H. 181 Schneider, H.P.G. 337, 436, 510 Schöffling, K. 464 Schönborn, J. 284 Schröder, K. 337, 436 Schwandt, P. 293 Scriba, P.C. 293 Serrano-Rios, M. 17 Shaar, C J . 187 Shapiro, D.J. 129 Siegmund, P. 158 Simon, T. 530 Singhai, R.L. 89 Sinha, M.K. 335 Sladek, Celia, D. 217 Slanina, P. 106 Smalstig, E.B. 187 Somogyi, J. 181 Sonka, J. 532 Stahl, M. 165 Stan, M. 12 Stan, M.A. 12 Stas,J. 532

XI

Steiner, H. 432 Story, J.A. 403 Strohfeldt, P. 167 Sulman, G. 92 Sundby, F. 425 Sutherland, DJ .B . 89 Sutter, B.Ch. 297 Sutter-Dub, M.Th. 297 Sutton, J. 428 Suvarnalatha, M. 417 Sverdük, R.C. 84,137,400 Swann, Margaret 129 Szende, L. 289

T Takrf, J. 289,530 Tal, El i . 92 Tamarit, J. 256 Tamarit, J. Jr. 256 Tamburrano, G. 447 Tarachand, U. 280, 491 Tashjian, A.H. J.R. 70 Tchobroutsky, G. 184 Thomas, J.A. 529 Thoreil, J. 79 Thum, Ch. 339

Tiengo, A. 245 Tjälve, H. 106 Torigoe, T. 334 Trautschold, I. 21 Tripathy, B.B. 528 Tschikof, R. 25 Tüllmann, A. 158 Tulloch, B. 525 Tulloch, B.R. 247, 357, 457 Tyhurst, M. 267

V Vallance-Owen, J. 85 Valori, C. 93 Vanhaelst, L . 190,506 Vargas, L. 275 Varotto, L. 202 Vegh, L. 429 Velasco, C. 427 Vincze, I. 181 Vinik, A.I. 95,396, 499, 526 Vogt, W. 293 Voigt, K . H . 36, 337,436 Vydelingum, N . 247, 357,457,

W Wajchenberg, B.L. 90 Walk, T. 170 Ward, W.F. 238 Weinges, K .F . 167,410,514 Wenzel, K.W. 169 Werner, S. 30, 319, 365 Westman-Naeser, S. 125 Whichelow, M.J. 234 Wickramasighe, R.H. 241 Willig, F. 181

Y Yamagata, S. 478 Yang, M.M.P. 430 Yoshimoto, Y. 343 Yudkin, J. 99

Z Zanisi, M. 432 Zbirkova, A . 532 Zebrowski, E.J. 121 Zermatten, A. 272 Ziegler, R. 243

XII

Biguanides 165 Bioassay, insulin 12 - , thyrotropin 9 Blood pressure, neutral red 111

SUBJECT INDEX

A A-cell, intracellular transport of

proteins 267 A-2-cell 49 Acetoacetate 256, 488 ACTH 1, 36, 226

effect on reabsorption of Cortisol 154

Addison's disease 17 Adenine nucleotides, pancreas 44 - , surface fluorescence 284 Adenohypophysis 208 Adenylate cyclase 208, 365, 443 - , islets of Langerhans 49,166 - , prostatic 89 Adipose Tissue 226, 357, 37G, 454 - and cells, rat - , human 229 - , protein kinase 146 - , rat 83 Adrenal cortex, mitochondria 241 Adrenalectomized dogs, neutral red

111 Adrenalectomy 30, 365 Adrenal gland, cat 414 Adrenaline 49, 234, 357 Adrenergic blocking agents 428 Adrenocorticotropic normo ne 1, 36,

154, 226 Age dependence, binding of testoste-

rone 213 Age, influence on pituitary 240 - , thyroid hormones 403 Albino rat 483 Allergie reactions 447 Alloxan 121 Alpha-cell, see also A-cell 392 Amiloride hydrochloride 115 Amino acid uptake, thyroid gland 196 Amino glutethimide administration

414 Anesthetics, local 106 Anterior pituitary hormones 337 - , simultaneous Stimulation test 436 Antibodies, insulin 175 Antigen-antibody complex 57 Anti-insulin serum 376 Antilipolysis 319 Antilipolytic activity 464 Antiovulatory activity 432 Antiserum heterogeneity 57 Arginine, chronic pancreatitis 95 - infusion 165 Artificial 0-cell 339 - pancreas 339 Ascorbic acid 234 Athyreosis 190 Atropine 115 Autoimmunity, endoerine pancreas

173

Autoradiography, thyroid slices 309

B 0-blockade 234 Beta-cell 392 "Big"-ACTH 1 - gastrin 1 - glucagon 1 - growth hormone 1

C Calcitonin 30, 319,410,517

human 161 - , lipolysis 243 - , porcine 161

ulcers 332 - , release 70 Calcium 70, 247, 319/329/365 - balance 62 - , calcitonin 30 - excretion, urinary 90 - , homeostasis 177 - , intracellular 525 - kinetics 62 Caloric intake 351 Calvarium 407 camp, see cyclic AMP Carbimazole 306 Carbohydrate metabolism 483 Carbon dioxide 457 Carbonic anhydrase 158 Cardiovascular changes, thyroid Dys-

funetions 93 Catecholamine excretion 532 Catecholamines 229, 414 - , levels in plasma and urine 93 - , release, neutral red 111 Chlorpropamide 386 Cholecystokinin, chronic pancreatitis 95 Cholesterol 403 - aeyltransferase 142 - , sidechain cleavage 241 Chronic pancreatitis 95 Circadian variations, carbohydrate toler-

ance 386 Citräte enzymes 264 Clomiphene ci träte, male 171 Coma, diabetic 339 Computer analysis, thyroxine distribu-

tion 202 Copper 329 Cortical cells 417 Corticosteroids 414 Corticosterone, diurnal rhythm 99 Cortisol 213,506 - , determination 429

influence of psychotropic drugs 5 30 - , plasma level 87 - , reabsorption site 154 - stop flow 154 Cortisone 365 - acetate 30 C-Peptide 1 Cross-reaction, HGH and HCS 80 Cyclic AMP 53, 70, 158, 208, 229, 247,

357, 365,457,529 - , excretion 314 - , fat cells of the rat 242

D Dehydroepiandrosterone, pregnancy 170 11-Deoxycorticosterone 241 2-Deoxy-D-glucose 21 Dexamethasone 303

DHA-sulfate, pregnancy 170 Diabetes 275 - , automatic treatment of acute condi-

tions 339 - coma 339 - , experimental 173 •-, genetic427 - , gonadotropins 121 - insipidus 289 - , rats430 - , streptozotocin 475 - , proinsulinlike material 439 Diaphragm 370 Diarrheic calves 434 Diazoxide, effect on islets 166 Diet 351 - and exercise, catecholamine excre­

tion 532 Dietary carbohydrates, diurnal rhythms

99 Diethylstilbestrol 522 Dihydroergotamine 303 Dihydrotestosterone 89 Diphenylhydantoin 475 Diphosphonate 62 Diuretic effect, glibenclamide 289 Diuretics 115 Diurnal rhythm 99 - variations, corticosterone and cations

329 D,L-ethionine 44 Dog 400 Dogs, hypothyroidism 137 Dopamine neuroleptics 187 Dwarfism 343 - , familial 80

E Exercise, thyroid hormones 403 Eminence median 94 Entero-insular axis 95 Enzymeactivities, muscle and liver 222 Epinephrine 158 Erythrocytes 158 Estradiol 213,217 Estrogen 208 Estrone 213 - tri tyrosine methyl ester, 1251-labelled

334 Estrous cycle 217 Ethanol, effect on glucogon secretion

245 17a-Ethynylestradiol in man 430 Ethynylestradiol, metabolites 333 Exercise, growth hormone secretion 184 - , muscular 488

F Fasting 229,499 Fat cells, free 12 - size 351 Fatty Acid 457 Fenfluramine 85, 86 Fettstoffwechselhormon 36 Fetus 376 F F A 137, 386, 400, 483, 488 Follicle stimulating hormone 121, 171,

335,336,514 Food deprivation 264

XIII

Guanidinobutyric acid 115 Guanidino-propionic acid 115 Guinea pig 267 Gut gli 396

Free Antigen 57 Free Fatty Acids 137, 386, 400, 483,

488 Fructose 370, 457 - , sulfonylureas 284 FSH 121,514 - and LH - urinary 336 - , diabetics 335 - , infertility 171

G Gastric secretion 517 - ulcer517

, restraint-induced 332 Gastrin 1, 260 Gastrointestinal hormones, effect of 2-

deoxyglucose 21 Gestation 376 Ghosts, fat cells 238 Glibenclamide 181, 289, 464 - , subcellular distribution 181 Glibornuride 386 Glucagon 1, 49, 70, 392, 443, 478 - , camel 425 - , effect on adenylate cyclase in islets

166 - , pancreatic 156, 396 - , paradoxical release 426 - secretion, neutral red 111 - pig 245 Gluconeogenesis 284, 381 Glucose 25, 234, 256, 319, 370, 376,

386, 392, 457, 470 - (3H)12

blood 115, 184 - controlled insulin infusion 339 - , diurnal rhythm 99 - homeostasis 392 - infusion test 400 - uptake, muscle 85, 86 Glucosyltransferase, kidney 427 Glucuronyltransferase 522 Gluconeogenesis 217 Glycemia, Addison's disease 17 - , neutral red 111 - Glycerol 483 ~ release 357 Glycogen 370, 457 - content, muscle and liver 222 Glycogenolysis 525 Glycogen synthesis 217 - synthetase 40, 457 Glycolysis 284 Glycolytic metabolites, pancreas 44 Goitre, nontoxic endemic and spora-

dic501 Gonadotropins, diabetics 121, 335 Group experiment, radioimmunoassay

for insulin 293 Growth hormone 1, 129, 275, 386,506 - , human 184, 303, 347 - , obese mice 125 - , propanolol-insulin Stimulation test

343 - , rat, radioimmunoassay 300 --, responses to physical exercise 428 - therapy 494 Growth rate 494 Guanidine Chloride, insulin secretion

115 Guanidinobutyramide 115

H HB 419181 HCG, 0-subunit 527 HCS 80 Hemicastration 417 Heparin 151 Hepatic disease, TSH-response 132 HGH, antibodies 484 - , immunological behaviour 80 3H-glucose 12 Hindlimb, isolated, rat 167 Histamine assay, TRF treatment 92 Histomorphometry 62 3 H-lipid counting 12 - extraction 12 Human chorionic gonadotropin (HCG)

527 - placental lactogen 347 Hydrocortisone treatment, Addison's

disease 17 0-Hydroxy butyrate 256 25-Hydroxycalciferol 534 Hyperbaric oxygen 40 Hypercholesterolemia 88 Hyperglycemic mice 40 Hyperinsulinemia 222 Hyperparathyroidism 314 Hyperthyroidism 93, 142, 190, 202, 400 - , cholesterol a cyltransferase 142 - , dogs137 - , treatment 92 Hypocalcemia, parathyroid response 91 - , insulin and glucagon 443 Hypophysectomie 5 3 - , protein kinase 146 Hypothalamic lesion 222 - obesity 370, 454 Hypothyroidism, primary 9

Insulin resistance 447 , obese mice 40,125

- , responsiveness 454 - RIA, group experiment 293 - secretion 177,470

, Addison's disease 17 in vitro, local anesthetics 106

, neutral red 111 , pancreozymin 21

- , sensitivity in normal rats 430 - tolerance test 84 - treated diabetes 184 Insulitis 173 Intestinal hormone 272 Intestine 407 Intravenous glucose, chronic pancreatitis

95 - glucose test, rapid 17 - tolbutamide test, Addison's disease 17 Iodide 306 - , hormonal 434 - trapping in vitro 309 IR-HGH 80 IRI, serum 137 Irreversible adsorbent 57 Islets, adenylate cyclase 166 - of Langerhans 49 Isolated fat cells 25 - islets 470 - pancreas 260 - perfused pancreas 256 K Ketoacidosis 339 Ketone bodies 53 Kidney 407

L Lactation 297 Lecithin 142 LH 514 - , diabetics 335

infertility 171 LHRF antibodies 94 LH-RH, see also LRH - , diabetic men 335 - , injection in lambs 240 - Stimulation 336 Lipase, hormone sensitive 247 Lipid counting (3H) 12 - extraction (3H) 12 - metabolism 347,483 Lipodystrophy 447 Lipogenesis 351 Lipolysis 226, 229, 247, 303, 351, 357 - , calcitonin 30 - , isolated fat cells 25 Lipolytic hormone 36 Lipolysis, influence parathyroid hormo­

ne and calcitonin 243 Lipoprotein lipase activity, heparin 151 Liver 407,491 - cirrhosis 142 - , enzyme activities 222 - , fructose metabolism 284 - glycogen 381 - membranes 443 - , metabolism 264 - , protein synthesis 280 - triglycerides 53 Local anesthetics 106 L R H , see also LH-RH 510

I Immunoassay, prolactin 499 Immunological specificity, HCG 527 Immunoreactive insulin (I.R.I.),

Addison's disease 17 Impotence 335 Infertility, male 171 Insulin 217, 219, 234, 247, 256, 272,

280, 357, 376, 386, 392, 443, 457 - administration 222 - antagonist 275 - antibodies 175, 447 - , bioassay 12 - , chronic pancreatitis 95 - , degradation by fat cell ghosts 238 - , diurnal rhythm 99 - , growth hormone 343 - , immunoreactive 400 - infusion, glucose-controlled 339 - , lactating rat 297 - , lipolysis 25 - , oscillations in response 423 - , plasma 115, 165, 184 - , receptors 242 - , recovery in radioimmunoassay 293 - release 526 - requirement 447

XIV

LRH, continuous administration 79 - , human males 79 Luteinizing hormone (LH) 121, 410 - releasing hormone (LH-RH) 410

M Magnesium 70, 329 - deficient rat 91 Malnutrition 82 Mannoheptulose 21 Microangiopathy 447 Monocomponent insulin (MC) 175, 447 Monospecies pork insulin (MS) 447 Monthly rhythms, hormones 506 Muscle 407 ~, enzyme activities 222 - , glucose uptake 85, 86

N Nafoxidine522 Nasal spray 510 Neuroleptics 187 Neurophysin I 83 Neutral red, insulin secretion 111 Newborn, human 392 Nicotinamide 280 Norepinephrine 226 Nutrition 226

O Obese hyperglycemic mice, glucagon

release 426 - , growth hormone levels 125 - mice 40 Obesity, hypothalamic 370 OGTT 137 Oligozoospermia, LRH 79 Oral amino acids load 272 - glucose 272

, chronic pancreatitis 95 , tolerance test, Addison's disease

17 Osmotic water flow, growth hormone

129 Ouabain 470

Oxygen consumption, liver 284

P Paget's disease of bone 62 Palmitate 256 Pancreas, metabolism 44 Pancreatectomized rat, partially 275 Pancreatectomy 53 Pancreatic a cell 267 Pancreatitis, chronic 95 Pancreozymin 21 - , chronic pancreatitis 95 Parathormone 1 Parathyroid hormone 243, 314 - , degradation 407 - , radioimmunoassay 407

respone to hypocalcemia 91 Pentobarbital 483 Pentose shunt 457 Perchlorate 306 Perfusion, rat lundlimb 167 Perifusion 470 Phentolamine, effect on islets 166 Phosphatase 247 Phosphate, inorganic 533 Phosphodiesterase 208 Phosphorylase 40 Pig 517

Pituitary, diabetic rats 121 - , "Fettstoffwechselhormon" 36 - function, malnutrition 82 - responses to LRH 79 - respone to LH-RH 240 Placenta 280 Plasma lipids, diurnal rhythm 99 Polydiethylstilbestrol phosphate 522 Posture, effect on Cortisol 87 Potassium 470 Pregnancy 297 - , DHA 170 Pregnant mice 491, 280 Procaine-HCL 357,457 Progesterone 217, 297, 430 Pro-hormones in tissues and in circula-

tion 1 Proinsulin 1 - antibody 439 - -like material 439 Prolactin 187,499,529 - and testosterone 529 - , human 347 - inhibiting factor (PIF) 187 - , thyroid diseases 190 Propranolol 53 Pro-parathormone 1 Propranolol, growth hormone 343 Prostaglandin Ei208 Prostaglandins 229 Prostate gland 529 Prostatic adenyl cyclase 89 Protein-binding assay, 25-Hydroxy-

calciferoi 534 Protein caloriec malnutrition (PCM)

528 - kinase 49, 208, 247

, hypophysectomy 146 - synthesis 196, 267, 488

, placenta and liver 280 Psychoactive drugs 187 Psychotropic drugs 530 PTH-fragment 1 Puberty, girls 325 Pyridine nucleotides, pancreas 44 Pyruvate 370, 457 - dehydrogenase 457

R Radioimmunoassay, data processing

293 - , quality control 293 - , thyrotropin 9

thyroxine 169 Rana hexadactyla 417 Rat, conscious 115 - , organs, gübenclamide distribu-

tion 181 Receptors 443 Red blood cells, potassium 526 Renal Cortisol reabsorption 74 - disease, TSH response 132 - tumors522 Rhythm, diurnal 99 Ribosomes 488

S S-adenosyl-methionine 333 Salmon calcitonin 161 Secretin 21, 272 - , chronic pancreatitis 95 Sex organs, diabetic rats 121 Sheep 0-lipotropin 303

Skeletal muscle 86 Sodium excretion, urinary 90 - transport, effect of growth hormone

on 129 Somatomedin 125, 319, 494 Steroids 417 - , urinary 325 Stüling cells 417 Streptozotocin 280, 475, 488 Stress 275 Suckling 264 - -food deprivation 381 Sugar, Blood 137,400 Sulfation factor 125 Sulfonylureas 284

Synthetic medium for perfusion 167

T T 3 190 TBG, competitive ligand-binding assay

435 - , estimation of 435 Testis compensatory hypertrophy 417 Testosterone 514, 529 - , and prolactin 529 - /cortisol 213 - , endogenous.88 Theophylline 303 Threshold, renal, cortisol 74 Thyrocalcitonin administration 533 Thyroglobulin synthesis 196 Thyroid 306 - activity 93 - , dog 309 Thyroidectomy 84 Thyroid gland, amino acid uptake 196 - hormone levels 528

s in nontoxic goitre 501 Thyroidism, asymptomatic 190 Thyroid secretion rate 403 - , sheep 309 - stimulating hormone (TSH) 410 Thyrotoxicosis 202 Thyrotropin 9, 506 - Releasing hormone 9, 499

, rat 432 (TSH-RH)410

Thyroxine 137, 202, 208 - (T 4 ) , old age 239 - , radioimmunoassay 169 Tissue culture, calcitonin secretion 70 Toad skin, growth hormone 129 Tolbutamide 386,464, 478 TRF92 TRH 190 - , hepatic and renal diseases 132 - in nontoxic goitre 501 Triamterene 429 Triglycerides 403 Triiodothyronine (T 3 ) , old age 239 Tryptic peptides and secretin 396 TSH 190

hepatic and renal diseases 132 - , serum, in nontoxic goitre 501 Tubulär maximum, cortisol 74 U,V Urinary excretion, sodium and calcium 90 Uncinate Dog 478 Vasopressin 1 W,Z Weanling rat 454 Zinc 329

293

Horm. Metab. Res. 6 (1974) 293-296 © Georg Thieme Verlag Stuttgart

Group Experiments on the Radioimmunological Insulin Determination*

I. Marschner1, P. Bottermann1, F. Erhardt1, R. Linke2, G. Löffler 1 , V. Maier3, P. Schwandt1, W. Vogt 1, and P.C. Scriba1

Sonderforschungsbereich 51 Muenchen, and Sonderforschungsbereich 87 Ulm (1 München, 2j utzing, ^uim), Germany

Summary

By means of two group experiments, performed in seven laboratories and using six different modifications in the in­sulin radioimmunoassay technique, it is shown that the use of Standards diluted in human serum of low basal insulin concentration, instead of buffer, improves appreciably the interlaboratory precision. The causes lies perhaps in some serum factor, which could disturb some modifications of the radioimmunoassay more than others. There was no im-provement of precision when the same insulin Standard was used in all laboratories. Standard curves made up in diffe­rent human fasting sera have a scatter only within the ränge of pipetting and counting error.

Key-Words: Group E x p e r i m e n t - I n s u l i n R I A - D a t a Pro­cessing - Q u a l i t y C o n t r o l - I n s u l i n Recovery

Introduction

Elimination of inter-laboratory differences of the results of clinical chemical analyses has recently drawn major attention. Quality control of routine methods largely is performed by means of partici-pation in group experiments. The aim of this work was to design and to test a modell for collabora-tive, comparative studies on the radioimmunological insulin determination with the goal of improving the inter-laboratory precision.

Material and Methods All experiments were performed on the same samples by seven individual groups of workers in different laboratories. First g r o u p e x p e r i m e n t ( 1 9 7 2 ) : For the first group experi­ment we dispatched the following two lots of serum samp­les to all seven partieipants.

Twentyfour samples from a serum pool, mixed from the serum of 5 patients, venipuneture one hour after breakfast.

Si> serum samples with different concentrations of insulin, taken from one patient during an oral glucose tolerance test (OGTT). The venipunetures were performed at 0, 5, 10 , 20 , 3 0 and 6 0 minutes after the administration of 100 gm glu­cose. Each sample was assayed in triplicate. In addition, each box with the frozen samples contained a vial of 2 nU insulin Standard preparation (kindly supplied by Fa. Buchler, Braunschweig - RCC, Amersham). With this insilin Standard each partieipant prepared a Standard curve

*Sipported by the Deutsche Forschungsgemeinschaft (SFB 5 1 , München, and SFB 87, Ulm).

Received: 15 Feb. 1974 Accepted: 12 Mar. 1974

in his own buffer system, and using his usual concentra­tions. The unknowns were then evaluated with the labora-tory's own Standard curve, as well as with the Standard curve containing the common insulin Standard. Second g r o u p e x p e r i m e n t ( 1 9 7 3 ) : The samples assayed in the second group experiment were as follows: Nine samples from a new serum pooL Triplicates of seven serum samples drawn from a further pa­tient during an OGTT. Times of venipuneture: 0, 5, 10, 15, 2 0 , 3 0 and 6 0 minutes after administration of glucose. Triplicates of 8 samples to form the recovery curve. These were made in human serum with a low basal concentration of insulin. For this we used the serum of a fasting patient which contained approximately 6 M U insulin/ml. 200 /utU insulin were then dissolved in each millilitre of an aliquot of this serum, so providing a stock serum Standard of 206 M U insulin/ml. From this, further serial 1 + 1 dilutions were made, using the rest of the low insulin serum as diluent. Thus the following Standards were obtained 206, 106, 56 , 3 1 , 18 .5 , 12 .3 , 9 .2 , and 6 /uU/ml. In all cases the identity of the samples was withheld from the partieipants.

Results

First group experiment The individual group modifications of the insulin RIA technique are shown in Table 1. Because all unknowns were measured in one assay-run, the with­in assay coefficient of Variation could be obtained from the 24 samples of the serum pool. This ranged between 3.4% and 14%. The scatter of the means of the 24 values from each laboratory was between 21 juU/ml and 46 /iU insulin/ml and had a coefficient of Variation .of 21%. The results from the OGTT-sera had a similar scatter. The means of the measured in­sulin concentrations lay between 13 juU (fasting) and 82 /iU/ml (60 minutes after glucose), and the coeffi­cient of Variation of 18% calculated in the lower con­centration ränge increased to 33% in the higher con­centration ränge. The result from the first group ex­periment was, that there was no improvement of inter-laboratory precision, when the same insulin Stan­dards were used by all groups.

Second group experiment Because of the different methods for evaluating RIA data (logit- or parabola-transformation, graphic-manual procedures), which intrinsically yield different hor­mone concentration values, particularly at either end of the Standard curve, all results were handled as raw counts in this second group experiment in contrast

294 I. Marschner et al.

Fig. l a Results of the insulin-determinations of the serum pool (n=9) as calculated from each laboratory's own Standard curve in buffer (second group experiment). The results from each laboratory are represented in a Gaussian-curve. As usual the peak of each curve represents the mean value, the width of each curve between upper and middle third is two Standard deviations. The peaks (means) have a scatter with a coefficient of Variation of 28%. The results of the first group experi­ment (cf. text) were similar. Fig. l b The same data, related to the recovery curve, i.e. Standards dissolved in low insulin serum. Coefficient of Variation of the means 8%.

to the first one. In this way we were able to evaluate all hormone concentrations using one method, and so record only the differences of the assay variations. This was done with a Computer program, using spline approximation to fit the Standard curve. A Siemens 404/3 data processing machine was employed. The curves were plotted by an on-line controlled Hagen-plotter (Marschner, E r h a r d t and Scriba 1973). The count rates of the unknowns from the various groups were related firstly to their corresponding Standard curve and then to their recovery curve.

Calculation o f the unknowns, using the S t a n d a r d curve i n buffer The means of the 9 serum pool samples from all seven groups had a scatter with a coefficient of Va­riation of 28%. This, and also the individual within assay precision were not appreciably different from

the results of the first experiment (Fig. la). Similar results were also obtained by comparing the hor­mone values from the OGTT in both group experi­ments (Fig. 2a).

C a l c u l a t i o n of the unknowns, using the recovery curve By calculating the insulin concentrations of the sam­ples from the serum pool (Fig. lb) and those from the OGTT (Fig. 2b) using the recovery curve (see above) we obtained a great improvement of inter-laboratory precision. Inspite of the fact, that the individual within-assay precision remained nearly un-influenced, the scatter of the means of the pooled serum was reduced to a coefficient of Variation of only 8% from 28%.

It was noticed, that the recovery curves in serum generally had a gradient steeper than the Standard curves in buffer (Fig. 3).

Table 1 Modifications of the radioimmunological determinations used by the partieipants.

Serum Volume Ml

Pre-Incubation h

Incubation

h

Temp.

°C

Pipetting Steps

B/F-Separation

A 100 6 24 4 3 Filtration on Cellulose Acetate Filters, B 100 6 18 4 3 Using Preprecipitated Antibody

C 100 - 17-24 4-8 2 Solid-Phase Technique Fa. Boehring;r

D 100 - 72 4 4 Ion Exchange Resin (Amberlite)

E 100 - 24 22 6 Sepharose-bound Antibody Pharmacia, Uppsala

F 50 3 8 22 3 Sephadex G-75-Column Chromatogaphy

G 100 - 120 37 3 Dextran Coated Charcoal Adsorption

Group Experiments on the Radioimmunological Insulin Determination 295

MICHROUNITS INSULIN/ML

120-h-

110»- •

100 J- •

MICROUNITS INSULIN/ML

120-

110--

100

Füg. 2a io MINUTES Fig. 2b io 00 MIN

Fig.. 2a Results of the insulin-determinations in the sera from the OGTT, related to each laboratory's own Standard curve in buffer. Ordinate: insulin concentration in pU/ml. Abscissa: time of venipuneture after glucose administration. The middle line* connects the means of all Single values of all laboratories, the shaded area covers two Standard deviations. Mean coeffi-ciemt of Variation 14%. Fig.. 2b The same data, related to the recovery curves: mean coefficient of Variation 6-5%. In the higher concentration ränge of the Standard curve no improvement was obtained (decreasing steepness).

We examined the usefulness of various human fasting sera for insulin recovery curves and prepared 7 re­covery curves as described above. Triplicates of 4 comtrol samples calculated by the Computer on each of the 7 recovery curves, showed a coefficient of Variation not greater than 7%, which lays within the precision of the assay.

Discussion

Group experiments are a well aeeepted procedure to examine the validity of laboratory results. For ordi-nary laboratory methods one or two determinations of a substance are sufficient for such a project where-as because of the nonlinear RIA Standard curves 6 to 8 determinations in triplicates of samples having con­centrations which are spread over the sensitive part of the Standard curve are essential.

It is desirable, that the concentrations of the samples are within a physiological context, like in our case from an OGTT.

To make sure, that the results represent all individu­al laboratory errors, each partieipant should handle his samples as in a routine assay. A detailed Informa­tion about the individual assay is not necessary. The only important fact is the agreement of results with those of other laboratories.

Since one is commonly interested in comparing as­say Systems and not the different evaluation proce-dures, it is advantageous to ask for count rates in-stead of hormone concentrations. From the obtain­ed count rates the hormone concentrations will be

8[%|

0

\ RECOVE RY C :URVE

STANDAR D CU RVE \ ^ \ ^

\ \

3.13 6.25 12.5 25.0 50.0 100.0 200.0

MICROUNITS INSULIN / ML

Fig. 3 A typical Standard curve in buffer and recovery curve. The latter has a steeper gradient. Ordinate: B / B 0

(B = antibody bound activity, B 0 = maximal antibody-bound activity in absence of cold hormone). Abscissa: logarithmic dose (/uU insulin/ml). The horizontal line at 87% is the limit of sensitivity, calculated as 3 Standard deviations of B0-values (n = 9).

296 I. Marschner et al.

calculated in a uniform fashion by an appropriate Computer program. From the count rates of the Standard curves one gets also Information about the steepness of the slope of the Standard curves and the limits of sensitivity (threefold Standard devia-tion of the zero-standard).

The two group experiments, regarded as pilot studies, show, that both the type of the RIA System (anti­body, bound-free-separation, incubation-time and -temperature, sample volume) are of secondary im-portance in improving inter-laboratory precision. It is, therefore, less important to standardize either of these. Differences in the purity of the tracer, the in-cubation and the antigen-antibody-reaction seem to play a greater role in serum samples than in pure buffer Systems. In view of this, it was considered better to dissolve the insulin Standard not in buffer, but in serum with a low basal insulin concentration. Comparable effects have been shown by E r h a r d t , Marschner, Pickardt, and Scriba (1973) in the meas-urement of hTSH.

A coefficient of Variation greater than 20% is con­sidered too high for satisfactory hormone measure-ment, particularly when needed frequently, for cli-nical and research purposes and when the RIA itself is relatively simple (Ashford et al. 1969). This can be improved considerably, however, by using Stan­dards in serum. In this way the inter-laboratory pre­cision approximates to the ränge of such assays as cholesterol or urea-nitrogen (Stamm and Büttner 1969).

It is apparent, that the precision will decrease a little, if each laboratory uses a different serum for its own Standards. Despite this, our results suggest, that this decrease would not be great. It is, furthermore re-commended that Standards be made by serial dilutions of 1 + 1, starting from one stock Solution containing about 200 to 300 juU/ml. Thereby, it is necessary to use only one pipette and one Single volume, resulting in a precision superior tc all other ways of pipetting Standard dilutions.

References

A s h f o r d , W.R., J . C a m p b e l l , J . K . D a v i d s o n A M . F i s h e r , R.E. H a i s t , A M . Lacey, B . L i n , J M . M a r t i n N i n a H . M o r l e y , K . Sudha R a s t o g i , a n d W.O. S t o r v i c k : A consideration of methods of insulin assay. Diabetes 18 (1969) 823-833

E r h a r d t , F . , I . M a r s c h n e r , Renate C. P i c k a r d t , P . C . S c r i b a : Verbesserung und Qualitätskontrolle der radioimmunolo­gischen Thyreotropin-Bestimmung. Z. klin. Chem. 11 (1973) 381-387

M a r s c h n e r , /., F . E r h a r d t , P . C . S c r i b a : The calculation of the radioimmunoassay Standard curve by "spline func-tion". Symposion on radioimmunoassay and related pio-cedures in clinical medicine and research. Internationa Atomic Energy Agency, Sept. 1973. Istanbul, Turkey

Stamm, £>., H . Büttner: Ringversuch zur Qualitätskontrolle 1968. Z. klin. Chem. 7 (1969) 393403

Requests for reprints should be addressed to: Dr. I. Marschner, Laboratorien für Klinische Chemie und Endokrinologie II. Medizinische Klinik der Universität D-8000 München 2, Ziemssenstr. 1 (Germany).