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Biomedical subjects

N R Lazarus

Publications and source records attributed to N R Lazarus.

At least 37 records · Page 2Linked to original sources
PubMed1978

The relation between insulin and adipocyte insulin receptors during treatment of human obesity.

Seventeen obese patients were submitted to 1 week's severe then 3 months moderate dietary restriction. The glycaemic and insulinaemic response to oral glucose was measured at baseline, after 1 week and at 3 months, and the results related to adipocyte insulin receptors estimated from fat biopsies taken on each occasion and to the weight lost. Adipocyte insulin binding was found to be significantly inversely correlated with the insulinaemic response to oral glucose (r = -0.54, P less than 0.05), an inverse relationship maintained even after weight loss had been achieved and displayed within as well as between subjects. Variation of 125I-insulin binding to adipocytes was related to receptor number and not to receptor affinity and could not be accounted for by differences in 125I-insulin degradation. Adipose cell size did not correlate significantly either with the insulinaemic response or with cell surface receptor density. The findings suggest: (1) that a significant proportion of obese subjects are not hyperinsulinaemic and that their metabolic response to weight reduction may differ from that of those who are hyperinsulinaemic; (2) that reduced insulin binding to adipocytes in the obese reflects hyperinsulinaemia, when present, and not adiposity as such; (3) that the degree of insulinaemia is a major determinant of the cell surface receptor density.

Adipose Tissue↗
PubMed1977

Reversal of streptozotocin-induced diabetes in rats by intraperitoneal implantation of encapsulated neonatal rabbit pancreatic tissue.

Implants of rabbit neonatal pancreas, encased in 'Nucleopore' chambers (0.4 micrometer) reversed streptozotocin-induced diabetes in the rat. Blood-glucose, plasma-insulin, and oral glucose-tolerance test returned to normal. An isolated, perfused, streptozotocin-treated pancreas removed from a diabetic animal did not secrete insulin and removal of implants after 6 weeks from six animals caused all animals to die in hyperglycaemia within 8 days. This shows that the implant did not lead to the re-establishment of endogenous pancreatic function. Implanted diced neonatal pancreas in three chambers removed after 6 weeks secreted glucagon, insulin, and pancreatic polypeptide in vitro. No rejection reactions were seen. Rabbit neonatal pancreatic implants may thus be feasible therapy in insulin-requiring diabetic patients. Implants of other non-syngeneic endocrine cells--i.e., pituitary, thyroid, and ovary--may be useful in other hypoendocrine syndromes.

Animals↗
PubMed1977

An in vitro system for studying insulin release: effects of glucose and glucose-6-phosphate.

1. Investigation of the ionic requirements of the in vitro insulin release system, which consists of cod islet plasma membrane and rabbit islet granules incubated at pH 6.5, showed that the presence of Ca(2+) was obligatory for the system to operate.2. Glucose-initiated insulin release was as effective in the presence of beta-gamma-methylene ATP, as it was in the presence of ATP. This analogue of ATP is a substrate neither for adenylate cyclase nor for any known animal membrane proteases. The effect of ATP on glucose mediated release is allosteric.3. Glucose (16 mM)-initiated insulin release was slower than that induced by glucose-6-phosphate (4 mM); 150 and 120 sec, respectively.4. The lag found with glucose-mediated insulin release was dependent upon glucose concentration. The lower the glucose concentration, the longer the lag. With 1 mM glucose the lag extended to 30 min.5. Once insulin release was initiated, the rate and amount of insulin release was independent of the glucose concentration.6. Pre-incubation of membranes with Ca(2+), glucose and ATP prior to the addition of granules, abolished the extended lag that had been obtained with 1 mM glucose. Events in the plasma membrane are the major contributor to the generation of the extended lag.7. The glucose analogue 5'thio-D-glucose, although not able to release insulin, was shown to compete with glucose for the glucoreceptor. By increasing the ratio of analogue to glucose the lag time increased. Thus, the lag time is dependent upon the ;effective' external glucose concentration.8. The max. amount of insulin released by 4 ng of membrane in the presence of glucose (16 mM) was 300 ng. The fact that membranes became refractory to glucose after this max. amount of insulin was released showed that recycling of release sites was not taking place in vitro and that granule: granule interactions were not occurring.9. The 120 sec lag before glucose-6-phosphate-initiated release was independent of glucose-6-phosphate concentration. The rate of insulin release with glucose-6-phosphate was concentration dependent.10. Glucose-6-phosphate did not cause further insulin release from a membrane that had released the max. amount of insulin it was capable of in the presence of glucose. The addition of tolbutamide (10 mM) to such a membrane did cause insulin release. This suggests that glucose and glucose-6-phosphate share a final common pathway.11. Adrenaline and somatostatin did not inhibit glucose-mediated insulin release.

Adenosine Triphosphate↗
PubMed1977

The ability of pancreatic polypeptides (APP and BPP) to return to normal the hyperglycaemia, hyperinsulinaemia and weight gain of New Zealand obese mice.

Intraperitoneal injections of avian pancreatic polypeptide (APP) and bovine pancreatic polypeptide (BPP) are capable of returning to normal the hyperinsulinaemia, hyperglycaemia and weight gain of New Zealand obese mice. The lag glucose tolerance also becomes indistinguishable from normal. The mechanism whereby these polypeptides cause reversion is not known. Reversion can also be brought about by the intraperitoneal implantation of islets from white mice into New Zealand obese animals. The implanted islets secrete mouse pancreatic polypeptide. We conclude that the New Zealand obese syndrome arises from a genetic lack of mouse pancreatic polypeptide. We suggest that in humans a lack of pancreatic polypeptide might manifest as a syndrome analogous to that found in New Zealand obese mice.

Animals↗
PubMed1976

The purification and biological properties of pancreatic big glucagon.

1. Big glucagon was present in extracts of ox, dog, rat and turkey pancreas, representing 10-15% of the glucagon immunoreactivity, and was shown to be of islet origin by its presence in extracts of isolated pigeon islets. 2. Big glucagon was homogeneous by immunoassay after polyacrylamide-gel electrophoresis and was more electronegative than little glucagon. 3. Big glucagon was purified from bovine pancreas, and its apparent molecular weight was estimated by gel filtration as 8200+/-9%. 4. Limited tryptic proteolysis of the molecule produced an immunoreactive component slightly smaller than little glucagon. 5. Linear dilution curves were obtained with mammalian big glucagons by using both enteroglucagon cross-reacting and 'little-glucagon-carboxyl-end-specific' antisera. 6. The half-times for the disappearance of the immunoreactivity of big and little glucagon that had been injected into the rat circulation were 6.9 and 3.2min respectively. 7. Big glucagon was approximately one-sixth as effective as little glucagon in displacing radioactive little glucagon from its liver membrane receptor. 8. Big glucagon was equipotent on a molar basis with little glucagon in the stimulation of the mouse islet adenylate cyclase, an indicator of insulinogenic activity. 9. On a molar basis, big glucagon inhibited basal liver adenylate cyclase activity to the same extent that little glucagon stimulated the enzyme. 10. Big glucagon was without effect on blood glucose concentration in the rat in doses up to 5mug/kg. 11. Big glucagon was equipotent, on a molar basis, with little glucagon in stimulating lipolysis in isolated chicken fat-cells.

Adenylyl Cyclases↗
PubMed1976

Studies on the biosynthesis of pancreatic glucagon in the pigeon (Columba livia).

The biosynthesis of glucagon was studied in microdissected pigeon pancreatic, islets. [3H]-Tryotophan and [3H]leucine were incorporated into big and little glucagon. No precursor-product relationship was evident between big and little glucagon after radioactive pulsechase and immunoreactive chase incubations. Radioactive and immunoreactive little glucagon and immunoreactive big glucagon were actively secreted and the synthesis of both glucagons was inhibited by high concentrations of glucose. [3H]Tryptophan and [3H]leucine were incorporated into an islet protein of about 20000mol.wt. Gel filtration of extracts of turkey pancreas revealed the presence of an immunoreactive peak of mol.wt. approx. 20000. This glucagon-immunoreactive component was also present in dog and ox pancreas and was stable to chaotropic agents and elution at various pH values. A similar-sized glucagon-immunoreactive species was present in the dog circulation. These results are discussed in the light of the presently accepted mechanisms of glucagon biosynthesis.

Animals↗
PubMed1976

An in Vitro system for studying insulin release caused by secretory granules-plasma membrane interaction: definition of the system.

1. An in vitro system is described in which insulin beta-granule-plasma membrane interaction can be studied. 2. The system shows an absolute requirement for physiological amounts of Ca2+ (2 muM) in order for insulin release to proceed. 3. ATP (5 muM) is abot to augment the Ca2+ effect. 4. Glucose (17 mM) alone does not cause insulin release but in the presence of Ca2+ is as effective as ATP. 5. When glucose, ATP and Ca2+ are added together a positive cooperative effect is produced with over 85% of the total insulin, added in the form of beta-granules, being released into the medium in 10 min. 6. The system responds to tolbutamide, in the presence of Ca2+ and ATP, by releasing insulin. 7. Diazoxide, a potent insulin inhibitor in vivo, demonstrates a similar activity in vitro. 8. Various control experiments utilizing alternative membranes, granules, nucleotides, sugars and phosphorylated intermediates of metabolism have all reinforced the specificity of the release mechanisms. 9. These results demonstrate that the in vitro system mimics responses found in the intact organism and can be utilized to dissect the mechanisms associated with exocytosis of insulin granules. 10. Preliminary experiments utilizing adrenaline granules-adrenal plasma membranes and pituitary granules-pituitary plasma membranes suggest that the in vitro system can be extended to all granule secreting processes.

Adenosine Triphosphate↗
PubMed1976

An approach to a molecular understanding of exocytotic insulin release.

By the use of an in vitro insulin releasing system, new insights into the meechanisms underlying the insulin exocytotic process have been gained. It is proposed that insulin release is initiated by glucose interacting with a glucoreceptor on the plasma membrane. Some properties of this receptor are discussed. It is postulated that after initiation of secretion, continued insulin release is under the control of phosphorylated intermediates of glucose metabolism, i.e. glucose-6-phosphate and phosphoenol pyruvate, operating via a membrane-bound protein kinase. The initiation of insulin release by glucose, and the augmentation of this initiation by the above mentioned intermediates, is viewed as a modified cascade system. The cascade theory of insulin secretion is postulated as an alternative to the threshold distribution hypothesis of insulin secretion. The action of tolbutamide in relation to the two pool theory of insulin secretion is discussed.

Animals↗
PubMed1975

The relation of conformation and association of insulin to receptor binding; x-ray and circular-dichroism studies on bovine and hystricomorph insulins.

Crystal and solution structure studies on insulins of different sequences and of widely different receptor binding affinities are reported. Bovine insulin, studied as a control, has a circular dichroism spectrum which is dependent both on protein concentration and zinc concentration. The spectrum appears to be related to the level of association of the insulin molecules. This implies that when using circular dichroism to compare solution structures of insulin derivatives or species variants one must make the comparison at equivalent levels of association and not merely at the same concentration. Changes in circular dichroism are related to the known crystal structure of zinc insulin hexamers. The chinchilla insulin spectrum shows a reduced zinc dependence in low-salt conditions which correlates with the inability to form crystals in similar conditions. This is attributed to an amino acid substitution at position B4. Crystals are obtained in high-salt conditions and X-ray diffraction patterns show these to be isomorphous with bovine 4Zn insulin crystals. Guinea pig insulin failed to crystallise under conditions which are normally conducive to the formation of crystals of zinc insulin hexamers and the circular dichroism showed no zinc dependence. This is consistent with a monomeric structure. The significance of the association behaviour of chinchilla and guinea pig insulins may be in the storage of the hormone in vivo. Whereas the monomeric form of chinchilla insulin has a structure closely related to bovine insulin, the circular dichroism indicates a gross structural difference for guinea pig insulin. This may be similar to that in des-A21, des-B30-insulin, as both lack the Arg-B22--Asn-A21 carboxylate ion pair. The similarity of structure of chinchilla and bovine insulins is reflected in their receptor binding whereas the low receptor binding of guinea pig insulin probably results from the changes in its conformation rather than an alteration in residues of a receptor binding region.

Animals↗
PubMed1975

Model for extrusion of insulin beta granules.

Considerable controversy surrounds the mechanisms of insulin secretion. This hypothesis attempts to explain the events which may lead to insulin extrusion through the beta-cell membrane. The role of various effectors of insulin secretion on the activities of membrane-bound enzymes is discussed. Glucose, without effect on these enzymes, did induce insulin secretion from insulin granules and plasma membranes in vitro. The model proposes a mechanism whereby glucose may trigger first-phase insulin secretion; reinforcing mechanisms, such as raised levels of cyclic adenosine monophosphate, could augment subsequent secretion.

Adenosine Triphosphatases↗
PubMed1975

The control of insulin receptors in the New Zealand obese mouse.

Insulin receptor concentrations in liver plasma membranes of New Zealand Obese (NZO) mice have been studied. When NZO mice were implanted with normal islets of Langerhans their blood glucose and plasma insulin declined. When the implanted islets were removed these changes were reversed and the mice reverted to their insulin resistant state. Changes were observed in the binding of 125I-insulin to liver plasma membranes of implanted NZO mice. Binding increased when the plasma insulin was decreased and conversely insulin binding decreased when the plasma insulin levels became elevated. The increased insulin binding was not accompanied by any changes in the affinity of NZO liver receptors for insulin.

Animals↗
PubMed1974

Insulin release from the perfused rat pancreas. Mode of action of tolbutamide.

Dextran-linked tolbutamide causes first-phase insulin release when perfused through the isolated rat pancreas. The linked sulphonylurea is also able to stimulate the membranal adenylate cyclase present in mouse islets. These facts make it highly likely that sulphonylureas exert their pharmacological action by interaction with the plasma membrane. Their action may be mediated via the adenylate cyclase enzyme.

Adenosine Triphosphate↗
PubMed1973

The biosynthesis of glucagon in perfused rat pancreas.

The biosynthesis of glucagon was studied by using the recirculated, isolated perfused rat pancreas. [(3)H]Tryptophan was initially incorporated into acid-ethanol-extractable protein, which on gel filtration was eluted with a molecular weight of about 9000 and contained a small amount of glucagon immunoreactivity. With longer incubation [(3)H]tryptophan incorporation into a second peak was obtained in an identical position with that of the majority of rat glucagon immunoreactivity. This peak of labelled protein exhibited migration characteristics on polyacrylamide-gel electrophoresis identical with those of rat glucagon and was identified as newly synthesized glucagon by demonstration of specific binding and dissociation behaviour with glucagon antibodies. The incorporation of [(3)H]tryptophan into acid-ethanol-extractable protein was inhibited by cycloheximide. High concentrations of glucose increased [(3)H]tryptophan incorporation into high-molecular-weight protein but decreased incorporation into proteins smaller than cytochrome c. The pattern of [(3)H]leucine incorporation into protein was similar to that of [(3)H]tryptophan.

Journal Article↗