Insulin

Secretion of Insulin.- A. The Kinetics of Insulin Release.- I. Introduction.- II. Dissociation of Different Phases of Insulin Secretion.- III. Influence of Non-glucose Stimulators.- IV. Role of Insulinogenesis.- V. Kinetic Characteristics of Insulin Response to Glucose.- VI. Theoretical Models for Insulin Secretion.- VII. Application in Man.- References.- B. Insulin Synthesis in ?-Cells.- I. Role of Proinsulin in Insulin Biosynthesis.- 1. Introduction.- 2. Chemical Structure and Properties of Proinsulin and Related Compounds.- 3. Secretory Cycle of the ?-Cell.- 4. Localization of the Converting Process.- 5. The Nature of the Proinsulin Converting Enzymes.- 6. The Products of Conversion and Their Intracellular Fate.- 7. Significance of Proinsulin for the Biosynthesis of Insulin.- II. Secretion of Proinsulin and C-Peptide and their Significance.- 1. Secretion of Proinsulin and C-Peptide In Vitro.- 2. Immunological Methods for the Determination of Circulating Proinsulin and Related Peptides.- 3. Secretion of Proinsulin and C-Peptide In Vivo.- a) Proinsulin.- ?) Normal subjects and animals.- ?) Pathological states.- b) Intermediate Forms.- c) C-Peptide.- 4. Peripheral Degradation of Proinsulin.- 5. Biological Activity.- a) In-Vivo Studies.- b) In-Vitro Studies.- 6. Prospects and Conclusions.- References.- C. Biochemistry and Biophysics of Insulin Secretion.- I. Energy Metabolism of the B-Cell.- 1. Introduction.- 2. Methodological Problems in the Study of Islet Metabolism.- a) Preparation of Islet Samples.- b) Microchemical Assay.- 3. Enzymatic Equipment and Nucleotide Levels of the B-Cells.- a) Phosphorylation of Glucose.- b) Glycolysis.- c) Pentose Phosphate Shunt.- d) Citric Acid Cycle and Respiratory Chain.- e) Amino Acids.- f) Lipids and Fatty Acids.- g) Glycogen Metabolism.- h) The Level of Nucleotides in the B-Cells.- 4. Substrate Utilization and Metabolism in the B-Cell.- a) Metabolism of Glucose.- b) Metabolism of Non-Carbohydrate Substrates.- 5. Energy Metabolism of the B-Cell In Relation To Insulin Biosynthesis and Secretion.- References.- II. Hexoses and Insulin Secretion.- 1. Introduction.- 2. Insulin Secretion Due to Hexoses and Pentoses as Well as Their Derivatives and Analogues.- a) Effects of Hexoses, Hexose Derivatives, and of Mannoheptulose.- ?) Stimulation of release.- ?) Inhibition of release by hexoses and mannoheptulose.- ?) Modulation of release by 2-deoxyglucose and mannoheptulose.- ?) Insulin release due to phlorizin.- ?) Effects of fasting and feeding on glucose-induced insulin release.- ?) The kinetics of release and of inhibition of release due to hexoses and mannoheptulose.- b) Effects of Polyols and Pentoses.- 3. The Permissive Action of Glucose Allowing Insulin Release Due to Calorigenic Molecules of Low Molecular Weight.- a) Glucose Dependency of Release Due to Various Stimuli.- b) Experimental Alterations of Chemosensitivity of Islets.- 4. Metabolic Function of Hexoses in the Islets of Langerhans.- a) Stimulation of Glycolysis and Respiration in Islets by Various Hexoses.- b) Dissociations of Metabolic and Insulin-Releasing Actions of Hexoses.- c) Dissociation of Metabolic and Permissive Functions of Glucose.- d) The Significance of Metabolite and Cofactor Levels in Islets Exposed to Glucose and Other Substrates.- 5. Interactions of Hexoses and Alloxan in Islets.- 6. Electrophysiological Effects of Hexoses on Islet Cells.- 7. A Possible Model Explaining the Multiple Actions of Glucose on the ?-Cells.- References.- III. Amino Acids and Insulin Secretion.- 1. Introduction.- 2. Transport and Metabolism of Amino Acids by Islet Cells.- a) Transport of Amino Acids.- b) Synthesis and Degradation of Amino Acids.- c) Protein Degradation.- d) Amino Acid Levels.- 3. Amino Acid-Induced Insulin Secretion.- a) L-Arginine and Related Substances.- b) L-Leucine and Related Substances.- c) Other Amino Acids.- 4. Conclusions.- References.- IV. Participation of the Adenylate Cyclase System.- 1. Introduction.- 2. The Regulation of cAMP Metabolism in the B-cell.- a) Membrane Receptors in the B-cell.- b) Insular Adenylate Cyclase.- c) Insular Phosphodiesterase.- d) The Level of cAMP in the B-cell.- ?) Immediate Regulation of cAMP Concentration in the B-cell.- ?) Long-term Regulation of cAMP Concentration in the B-cell.- e) Conclusion.- 3. The Effect of cAMP Upon Insulin Release.- a) The Insulinotropic Action of cAMP.- b) Combined Effects of cAMP and Other Insulino tropic Agents.- c) Combined Effects of cAMP and Inhibitory Agents.- 4. The Mode of Action of cAMP in the B-cell.- a) Effects of cAMP on Glucose Metabolism in the B-cell.- b) Effects of cAMP on Insulin Biosynthesis.- c) Effects of cAMP on Islet-cell Protein Phosphokinase.- d) Effects of cAMP on Tubulin Metabolism in the B-cell.- e) Effects of cAMP on Calcium Handling by the B-cell.- 5. Physiological Significance of the B-cell Adenylate Cyclase System.- a) Insulinotropic Action of Hormonal Polypeptides and the Entero-Insular Axis.- b) Inhibition of Insulin Release by Catecholamines During Stress and Exercise.- 6. Concluding Remark.- References.- V. Role of Cations.- 1. Introduction.- 2. Calcium.- a) Effect of Calcium Upon Insulin Release.- b) The Handling of Calcium by the B-cell.- c) The mode of Action of Calcium in the B-cell.- 3. Barium.- 4. Magnesium.- 5. Sodium.- a) Effect of Sodium on Islet Function.- b) Insular Handling of Sodium.- 6. Potassium.- a) Effect of Potassium on Islet Function.- b) Insular Handling of Potassium.- 7. pH.- a) Effect of pH on Islet Function.- b) Intracellular pH of Insular Tissue.- 8. Cations and Secretory Granules.- 9. Conclusion.- References.- VI. The Bioelectrical Activity of the Islet Cell Membrane.- 1. Introduction.- 2. Measurement of Electrical Properties of Islet Cells.- 3. Electrical Activity Induced by Islet Stimulants.- 4. Relationship Between Electrical Activity and Insulin Release.- 5. Effect of Ions on Glucose-Induced Electrical Activity.- a) Monovalent Ions.- b) Divalent Cations.- 6. Effects of Inhibitors and Anoxia on Electrical Activity.- 7. Effect of Diabetogenic Agents.- 8. Discussion.- References.- D. Effects of Sulfonylurea Derivatives on Pancreatic ?-Cells.- I. Introduction.- II. Descriptive Aspects of Insulin Release, Insulin Biosynthesis, and ?-Cell Morphology.- 1. Acute Effects on Insulin Release.- 2. Insulin Biosynthesis and Long-term Effects on Insulin Secretion.- 3. ?-Cell Morphology.- III. Explanatory Hypotheses Concerning the Insulin-Releasing Action of Sulfonylureas.- 1. Enhanced Glucose Recognition.- 2. Enhanced Recognition of Amino Acids.- 3. Increase of Cyclic AMP.- 4. Redistribution of Metal Ions.- 5. Binding to Plasma Membrane.- 6. Direct Effects on Insulin Storage.- IV. Concluding Remarks.- References.- Pharmacokinetics of Insulin.- A. Distribution in the Organism.- I. Introduction.- 1. Early Distribution Studies.- 2. Specificity of Distribution Pattern of 131I-labelled Insulin.- 3. Limitations of Early Distribution Studies.- 4. Reinvestigation of Distribution of 131I-labelled Insulin in the Rat.- 5. Role of the Liver and Kidneys in the Distribution and Degradation of Insulin.- 6. Intracellular Distribution of Insulin.- II. Dynamics of Insulin Distribution.- III. Kinetics of Insulin Distribution.- 1. Rate of Plasma Insulin Disappearance (Half-Life Concept).- 2. Limitations of Plasma Half-Life Estimates.- 3. Alternative Methods of Analysis of Plasma Insulin Disappearance.- 4. Limitations of Estimation of Metabolic Clearance Rates or Fractional Loss Rates of Insulin from Plasma.- IV. Summary and Conclusions.- References.- B. Degradation of Insulin.- I. Qualitative Aspects of Inactivation of Insulin.- 1. Inactivation of Insulin by Tissue Brei and Extracts.- 2. 131I-Insulin Degradation by Tissue Extracts, Slices and Homogenates.- 3. Specificity of 131I-Insulin Degradation by Tissue Extracts and Homogenates.- 4. Further Studies on the Nature, Specificity, and Localization of Enzymatic Degradation Processes for Insulin.- a) Evidence for Reductive Cleavage of S-S Bonds in Insulin.- b) Evidence for Proteolytic Degradation of Insulin.- II. Subcellular Localization of Insulin Degradation.- III. Regulation of Insulin-Degrading Activity.- IV. Quantitative and Kinetic Aspects of Proteolytic Degradation of 131I-Insulin.- V. Comments.- VI. Summary and Conclusions.- References.- Effects of Insulin and Proinsulin.- A. Insulin Receptor Interactions and the Action of Insulin.- I. Introduction.- II. Insulin Receptors.- 1. Binding of Iodoinsulin to Receptors.- 2. Localization of Receptors.- a) Localization of Receptors to Cell Surfaces.- b) Assymetric Positioning of the Insulin Receptor in the Membrane.- 3. Insulin Receptor Interaction.- a) Properties of the Insulin Receptor Interaction.- b) Effect of Enzymatic Digestions.- ?) Neuraminidase.- ?) Proteases.- ?) Galactosidases.- ?) Phospholipases.- c) Effects of Insulin-Agarose Derivatives.- d) Insulin Receptors and Plant Lectins.- e) Insulin-Resistant States.- 4. Solubilization and Isolation of the Insulin Receptor.- III. Mechanism of Insulin Action.- References.- B. Effects of Insulin on Cellular Protein Synthesis.- I. Introduction.- II. Insulin and Amino Acid Transport in Muscle Reconsidered.- 1. Relation of Insulin Action on Protein Synthesis to the Action of the Hormone on Amino Acid Transport.- 2. Insulin and Amino Acid Transport in Muscle.- a) Accumulation of Individual Natural Amino Acids.- b) Amino Acid Transport in the Absence of Protein Synthesis.- 3. The Nature of the Effect of Insulin on Amino Acid Transport.- 4. Does Availability of Amino Acids Limit Protein Synthesis in Muscle ?.- 5. The Dilemma of the Functional Heterogeneity of the Intracellular Amino Acid Pool.- a) Evidence for Compartmentalization.- b) Possible Organization of Amino Acid Pools.- c) Amino Acid Pools and the Measurement of Protein Synthesis.- III. Insulin, Diabetes and the Function of Muscle Ribosomes.- 1. Muscle Ribosomes from Diabetic Animals Catalyze Protein Synthesis Less Effectively than Do Ribosomes from Normal Animals.- 2. Small Amounts of Insulin Administered to Diabetic Animals Rapidly Increase the Synthesis of Protein by Ribosomes.- 3. Insulin Increases the Synthesis of all Muscle Proteins.- 4. Diabetes Reduces the Number of Active Ribosomes in Muscle.- 5. Preparations of Ribosomes from the Muscle of Diabetic Animals Contain Fewer Polysomes and More Monomers than Normal.- 6. Insulin-Induced Formation of Polysomes and Increase in Protein Synthesis Do not Require the Synthesis of RNA.- 7. The Reduced Ability of Diabetic Ribosomes to Translate Messenger RNA is not Related to the Binding of the Template.- 8. Diabetic Ribosomes Are Less Effective than Normal in the Translation of Poly- uridylic Acid at Lower Concentrations of Magnesium and More Effective at Higher Concentrations of the Cation.- 9. When Hybrid Ribosomes Containing a Normal and Diabetic Subunit Are Constructed, the Defect in Protein Synthesis Appears to Be Carried by the 60S Subunit.- 10. Diabetes Does not Alter the Peptidyl Transferase Activity of Ribosomes.- 11. The Elongation Factor-2 Catalyzed Hydrolysis of GTP by Muscle Ribosomes is not Changed in Diabetes.- 12. Diabetic Ribosomes Bind Less Phe-tRNA than Normal at Low Concentrations of Magnesium and More at High Concentrations.- 13. Diabetic Ribosomes Are Less Effective than Normal in the Translation of Polyuridylic Acid and RNA Extracted from Encephalomyocarditis Virus (EMCV RNA).- 14. Formation of an Initiation Complex by Normal and Diabetic Ribosomes.- IV. Insulin, Diabetes, and the Structure of Ribosomes.- 1. No Difference Has Been Found in the Proteins of Normal and Diabetic Ribosomes.- 2. No Difference Has Been Found in the RNA of Normal and Diabetic Ribosomes, although Diabetes Does Decrease the Number of Ribosomes to Be Found in Muscle.- 3. Diabetes Decreases the Melting Temperature of Ribosomes.- 4. The Pleotypic Program and the Intracellular Mediator of Insulin Action.- V. Coda.- References.- C. Effects of Insulin on Nucleic Acids, Nucleotides and Cyclic AMP.- I. Introduction.- II. Preliminary Considerations.- 1. Insulin as an Inductor or Depressor.- 2. Insulin as a Ligand Interacting with the Cell Membrane.- III Action of Insulin on Nucleic Acids.- 1. In Rat Diaphragm.- 2. In Liver.- 3. Discussion.- IV. Action of Insulin on the Incorporation of 32P-Labelled Inorganic Phosphate into Mononucleotides in Rat Diaphragm and Adipose Tissue.- 1. Experimental Approach.- 2. Phosphate Turnover in the Diaphragm of the Normal Rat.- 3. Phosphate Turnover in the Diaphragm of the Hypophysectomized Rat Treated with or without Growth Hormone.- 4. Phosphate Turnover in Isolated Adipose Tissue Cells.- 5. Effects of Insulin on Phosphate Turnover.- a) Discussion.- b) Significance.- V. Interaction of Insulin with the Cyclic AMP System.- 1. The Action of Insulin upon the cAMP Level.- 2. Mechanisms of the Effect of Insulin upon the cAMP Level.- a) Decrease in the Activity of Adenylate Cyclase.- b) Increase in the Activity of Cyclic AMP-Phosphodiesterase.- c) Is Insulin Acting at the Kinase Level ?.- 3. Conclusion: The Search for “Another” Transducer Specific for Insulin.- VI. General Conclusions.- References.- D. Metabolic Effects on Muscular Tissue.- I. Introduction.- II. Muscular Tissues Used for the Study of Insulin Action.- III. Effects of Insulin on Transport Processes.- 1. Glucose and Other Sugars.- a) Use of Sugars and Analogues.- b) Methodical Problems.- c) Insulin Membrane Action and Degradation.- d) Kinetics.- e) Inhibition of Insulin-Stimulated Sugar Transport.- 2. Electrolytes and Water.- a) Monovalent Cations.- ?) Potassium.- ?) Sodium.- b) Divalent Cations.- c) Anions.- d) Membrane Potential.- e) Water.- 3. Relationship Between the Transport of Glucose and Electrolytes.- a) Extracellular Ionic Milieu.- b) Intracellular Ionic Milieu.- c) Ionic Permeability of the Plasma Membrane.- IV. Effects of Insulin on Metabolic Processes.- 1. Sugar Phosphorylation.- 2. Glycogen Metabolism.- a) Amount and Labelling Studies.- b) Glycogen Synthetase.- 3. Glycolysis.- 4. Oxidation of Glucose and Other Substrates.- 5. Lipid Metabolism.- 6. Electrolytes and Glucose Metabolism.- V. “Insulin-Like” Effects.- 1. Contractile Activity.- 2. Anoxia and Metabolic Poisons.- 3. Hyperosmolarity.- 4. Enzymes.- 5. Compounds Structurally Related to Insulin.- References.- E. Metabolic Effects on Adipose Tissue in vitro.- I. Introduction and Early Studies.- II. Description of the Effects of Insulin.- 1. Membrane Transport of D-glucose and Related Sugars.- a) Recognition of Sugar Transport as a Major Locus of Insulin Action.- b) Evidence that the Transport of D-glucose is Accelerated by Insulin.- c) Transport Studies in Ghosts.- d) Characteristics of the Effect of Insulin on D-glucose Uptake.- e) Effect of Insulin on the Transport of Other Sugars.- f) Role of Monovalent Cations in Sugar Uptake.- 2. Glycogen Metabolism.- 3. Fatty Acid Synthesis.- a) Conversion of Intracellular Glucose to Pyruvate.- b) Conversion of Pyruvate to Intramitochondrial Citrate.- c) Conversion of Intramitochondrial Citrate to Fatty Acids.- 4. Triglyceride Metabolism.- a) Esterification of Fatty Acids.- b) Hydrolysis of Triglycerides.- c) Uptake of Triglycerides.- 5. Tissue Respiration and ATP Production.- 6. Amino Acid Metabolism and Protein Synthesis.- III. Analysis of the Mechanism of Action of Insulin.- 1. The Insulin Receptor.- 2. Correlation with Tissue cyclic-AMP Concentration.- 3. Role of Adenylate Cyclase.- 4. Role of Phosphodiesterase.- 5. Reflections.- References.- F. Direct In Vitro Effects of Insulin on Liver Metabolism.- I. Effects of Insulin on the Net Glucose Balance and on Glycogen Metabolism.- II. In Vitro Effects of Insulin on Gluconeogenesis in Liver.- III. The Mode of Action of Insulin on 3?,5?-cAMP-Mediated Processes in Vitro.- IV. Effects of Insulin on Electrolyte Metabolism.- V. Effects of Insulin on Hepatic Lipid Metabolism.- VI. Effects of Insulin on the Metabolism of Amino Acids and Proteins.- References.- G. Action of Insulin on Some Other Organs and on Differentiation.- I. Action on Stomach Function and Exocrine Pancreas.- 1. Action on Stomach Function.- a) Stimulation of Gastric Secretion by Insulin Hypoglycemia.- b) Inhibitory Actions of Insulin.- c) Direct Mucosal Stimulation of Acid Secretion.- d) Participation of Intracellular Mediation in Oxyntic Cells.- References.- 2. Action on the Exocrine Pancreas.- a) Influence of Insulin on Exocrine Pancreas Secretion.- b) Influence of Insulin on Synthesis of Pancreatic Juice Components.- c) Trophic Effects of Insulin on Exocrine Pancreas Tissue.- References.- II. Action on the Mammary Gland.- 1. General Metabolic Alterations.- 2. Specific Functional Alterations.- References.- III. Action on the Crystalline Lens.- 1. Diabetic and Sugar Cataract.- 2. Hexose Transport in Lens.- 3. Lens Differentiation.- References.- IV. Action on Some Mesenchymal Tissues and Single Cells.- 1. Action on Fibrocytes and Mesenchymal Tissues.- a) Action of Insulin on Fibroblasts in Culture.- b) Action of Insulin on Skin and Wound Tissue.- c) Action of Insulin on Cartilage and Bone.- References.- 2. Action on Tumor Cells.- References.- 3. Action on Blood Cells.- a) Erythrocytes.- b) Granulocytes and Lymphocytes.- References.- H. Metabolie Alterations in the Body by Insulin.- I. Effect of Insulin on the Fate of Glucose.- 1. Introduction.- 2. Isotope Dilution Techniques.- a) Definitions and Problems Arising from Incomplete Intermixing.- b) Recycling of Label.- c) Data from Studies Applying Isotope Dilution Techniques.- ?) Changes in the fate of body glucose induced by a deficient supply of insulin or by the diabetic state.- ß) Changes in the fate of glucose induced by insulin.- ?) Effect of glucose loading on the fate of body glucose as measured by isotope dilution.- 3. Effect of Insulin on the Fate of Body Glucose as Measured by Catheterization Techniques.- a) Measurements of the Net Balance of Glucose across the Liver.- b) Effect of Insulin on Net Splanchnic Glucose Output.- 4. Effect of Insulin on Glucose Utilization by the Extrahepatic Tissues.- 5. Conclusions.- References.- II. Alterations in Fat Metabolism.- 1. Introduction.- 2. Normal Metabolism of Lipids.- a) Postprandial State.- b) The Postabsorptive State (12-18 h after the Last Meal).- 3. Effects of Severe Insulinopenia.- a) Short-Term Insulin Deficiency (up to 6 h).- b) Long-Term Insulin Deficiency (One Day or More).- 4. Glycogenesis Type I.- 5. The Primary Lipodystrophies.- 6. Excess Insulin and Endogenous Hypertriglyceridemia.- 7. Endogenous Hyperlipemia and Insulin Resitance.- 8. Conclusions.- References.- I. Modification of the Effects of Insulin by Hormonal Factors.- I. Introduction.- II. Insulin.- 1. Effect on Blood Glucose Concentration.- 2. Effect on Hepatic Glucose Output.- 3. Effect on Perfused Liver.- 4. Effect of Endogenous Insulin on Hepatic Glucose Output.- 5. Effect on Glycogen Concentration.- 6. Effect on Hepatic Gluconeogenesis.- 7. Effect on Glucose Uptake.- 8. Comments.- III. Growth Hormone.- 1. Hypophysectomy — Effect on Glucose Production, Uptake, and Sensitivity to Insulin.- 2. Growth Hormone Administration in the Hypophysectomized Animal.- 3. Growth Hormone Administration in the Normal Animal.- 4. Growth Hormone-Induced Diabetes.- 5. Comments.- IV. Adrenal Glucocorticosteroids.- 1. Adrenalectomy — Effect on Glucose Production, Uptake, and Sensitivity to Insulin.- 2. Glucocorticoid Administration in Hypophysectomized and Adrenalectomized Animals.- 3. Glucocorticoid Administration in the Normal Animal.- 4. Comments.- V. Epinephrine.- 1. Effect on Hepatic Glucose Output.- 2. Effect on Glucose Uptake.- 3. Comments.- VI. Glucagon.- 1. Effect on Hepatic Glucose Output.- 2. Effect on Glucose Uptake.- 3. Comments.- VII. Prolactin and Human Placental Lactogen (HPL).- VIII. Thyroid Hormones.- IX. Concluding Remarks.- References.- Immunopathology of Insulin.- A. Insulin Allergy, “Cellular” Antibodies and Insulitis.- I. Insulin Allergy.- 1. Introduction.- 2. Clinical Symptoms.- a) Local Delayed Reaction.- b) Immediate Local or Generalized Reactions.- c) Other Types.- 3. Frequency.- 4. Diagnostic Procedures.- a) Skin Tests.- b) Laboratory Findings (Cytological and Serological Studies).- ?) Delayed allergic reaction.- ?) Immediate type allergy.- c) Histological Examinations.- 5. Pathogenesis and Classification.- 6. Concepts of Etiology.- a) Immunogenicity of Insulin and the Role of Proinsulin and Insulin-Related Proteins.- b) Antigenic Determinants of the Insulin Molecule.- 7. Relationship to Insulin Resistance.- 8. Relationship to Lipodystrophy.- 9. Treatment.- a) Delayed Allergic Reaction.- b) Immediate Allergic Reactions.- ?) Hypoglycemic agents.- ?) Change of insulin.- ?) Desensitization.- ?) Other measures.- II. “Cellular” Antibodies and Insulitis.- References.- B. Insulin-Binding Antibodies of Serum and Insulin Resistance.- I. Methods for the Determination of Insulin Antibodies.- 1. Determination by Inhibition of Insulin Effects.- a) Inhibition of Insulin Effects in Animals.- b) Inhibition of Insulin Effects in Isolated Tissue.- c) Inhibition of Insulin Degradation and Elimination.- 2. Determination of Insulin-Binding Antibodies by Immunological Tests.- a) Serologic Methods.- b) Passive Antibody Transfer.- 3. Detection of Insulin Antibodies by Direct Measurement of Insulin Antibody Binding.- a) Electrophoresis.- b) Ultracentrifugation.- c) Precipitation of Insulin-Protein Complexes.- d) Adsorption Techniques.- 4. Calculation of Quantitative Parameters of Insulin-Antibody Binding at Equilibrium.- II. Chemical-Physical Properties of Insulin Antibodies.- 1. Classification in Regard to Serum Protein Fractions.- 2. Two Main Components of Insulin Antibodies of IgG Class.- III. Influence of the Insulin Molecule on its Binding to Insulin Antibodies of Serum.- 1. Effect of Species Differences on Insulin Immunology.- 2. Effects of Chemical Modifications on the Antibody Binding of Insulin.- a) Insulin Chains and Chain Fragments.- b) Insulin Modifications with Intact Spatial Configuration.- ?) Analogs of insulin with shortened chains.- ?) Insulin analogs with prolonged chains.- ?) Insulin analogs with varied primary structure.- ?) Insulin derivatives.- ?) Cross-linked insulins.- ?) Insulin with partially cleaved disulfide bridges.- IV. Insulin Antibodies in Man.- 1. Without Previous Insulin Treatment.- 2. After Insulin Treatment.- a) Insulin-Binding Antibodies and Insulin Requirement.- b) Insulin-Binding Antibodies and the Quality of Metabolic Control in Diabetics.- c) Acute Effects of Insulin Antibodies on Metabolic Processes.- d) Chronic Effects of Insulin-Binding Antibodies on Metabolic Processes.- V. Antibody-Dependent Insulin Resistance.- 1. Pathogenesis.- 2. Frequency of Occurrence.- 3. Clinical Picture.- 4. Therapy.- References.- Determination and Preparations of Insulin.- A. Radioimmunoassay of Insulin.- I. Introduction.- II. Principles of the Radioimmunological Method.- III. Methods of Radioimmunoassay of Insulin.- 1. Materials.- 2. Insulin Labeling.- a) Principles.- b) Method of Labeling Insulin.- c) Purification of Iodine-Labeled Insulin.- 3. Separation of Free Insulin from Antibody-Bound Labeled Insulin by Means of Amberlite.- 4. Sensitivity and Accuracy of the Radioimmunoassay of Insulin.- a) Serum Dilution and Antiserum Used.- b) Concentration of Radioactively Labeled Insulin.- 5. Serum Insulin Assays.- 6. Other Methods of Separation.- a) General Survey.- b) Double-Antibody Method.- c) Separation with Charcoal and Dextran.- d) Cellulose Powder.- 7. Use of Insulin Receptors for Assay Purposes.- IV. Assay of Proinsulin and C-Peptide.- 1. Proinsulin Assay.- 2. C-Peptide.- References.- B. The Biological Assay of Insulin-Like Serum Activity (ILA).- I. Introduction.- II. Principle and Survey of Insulin Bioassays.- 1. In- Vivo Methods.- 2. In- Vitro Methods.- a) Rat-Diaphragm Method.- b) Rat-Adipose-Tissue Method.- c) Rat-Fat-Cell Method.- III. Practical Execution of Insulin Bioassays.- 1. In-Vivo Methods (Glycogen Synthesis of the Rat Diaphragm in Situ).- a) Experimental Animals.- b) Method.- c) Analytic Methods.- d) Calculation.- e) Norms.- f) Interference.- 2. In- Vitro Methods.- a) Determination of Insulin Activity from Glycogen Synthesis of the Isolated Rat Diaphragm.- ?) Experimental animals.- ß) Materials.- ?) Method.- ?) Analytic methods.- ?) Calculation.- b) Determination of the Insulin Activity from 14CO2 Production of Isolated Rat Epididymal Adipose Tissue.- ?) Experimental animals.- ß) Materials.- ?) Method.- ?) Analytic methods.- ?) Calculation.- ?) Norm.- IV. Forms of ILA in the Blood and their Significance: Conclusions.- 1. Immunologic and Biologic ILA.- 2. Nonsuppressible Insulin-like Activity (NSILA).- 3. Bound Insulin.- 4. Synalbumin.- References.- C. Methods for the Standardization of Insulin and the Determination of Blood Glucose Concentration.- I. Standardization of Insulin.- 1. Synopsis and Introduction.- 2. Methods of Standardization.- a) Mouse Convulsion Test.- b) Hypoglycemic Effect in Rabbits.- c) Standardization of Insulin Preparations with Prolonged Action.- d) Statistics.- II. Methods of Determining Blood Glucose Concentration.- 1. Chemical Methods of Measuring Blood Glucose Concentration.- a) Methods Based on Reduction.- b) Formation of Osazone.- c) Furfural-Reaction.- 2. Enzymic Methods of Determining Glucose.- a) Digestion by Microorganisms.- b) Reaction with Pure Enzymes.- ?) Glucose oxidase plus peroxidase.- ß) Glucose oxidase without peroxidase.- ?) Hexokinase plus glucose-6-phosphate-dehydrogenase.- References.- D. The Isolation of Insulin from the Pancreas.- I. General Aspects of Insulin Isolation.- 1. The Need for Insulin and How It Is Satisfied.- 2. Properties of the Pancreas Influencing Isolation.- 3. Properties of Insulin Influencing Isolation.- 4. Methods of Detecting Impurities.- II. Methods of Insulin Isolation.- 1. Extraction and Precipitation.- 2. Adsorption to Ion Exchangers.- 3. Table: Yield of Insulin Obtained Using Different Methods.- III. Insulin Purification Using Chromatographic Methods.- 1. Paper Chromatography.- 2. Gel Chromatography.- 3. Ion Exchange Chromatography.- a) Cation Exchangers.- b) Anion Exchangers.- IV. Special Purification Processes.- 1. Countercurrent Distribution and Partition Chromatography.- 2. Electrophoresis.- V. Table: Isolation of Insulin from Different Species.- VI. Isolation of Insulin Analogs and the C-Peptide.- References.- E. Insulin Preparations with Prolonged Effect.- I. Introduction.- 1. History.- 2. Pharmaceutical Chemistry.- 3. Absorption.- 4. Timing.- 5. Mixtures.- 6. Stability.- 7. Immunological Side Effects.- II. Insulin Preparations.- 1. PZI and NPH.- a) Protamine Insulin.- b) PZI.- c) NPH.- d) Pharmaceutical Composition.- 2. Surfen Insulins.- 3. The Lente Insulins.- a) Introduction.- b) Chemistry.- c) Crystallization of Insulin.- ?) The role of metal ions.- ?) The shape and size.- d) Composition.- e) Timing of Action.- f) Clinical Results.- 4. Insulin Rapitard.- III. Monocomponent Insulin.- 1. Introduction.- 2. Immunogenicity of Components of Crystalline Insulin in Rabbits.- 3. Preparation and Stability of MC Insulin Preparations.- 4. Clinical Results.- a) Insulin Antibodies.- b) Antibodies Against Proinsulin and a-Component.- References.- Author Index.