Biomedical subjects

A Sener

Publications and source records attributed to A Sener.

At least 181 records · Page 10Linked to original sources

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release. Functional response of islets to L-arginine and L-ornithine.

L-Arginine and L-ornithine stimulate insulin release from pancreatic islets exposed to D-glucose. This coincides with an increased outflow of 86Rb and 45Ca from prelabelled islets and an increased net uptake of 45Ca by the islets. In the presence of D-glucose, L-lysine stimulates insulin secretion to the same extent as L-arginine or L-ornithine, but the hormonal release is not further enhanced by combinations of these cationic amino acids. L-Arginine or L-ornithine failed to enhance insulin release evoked by either L-leucine or 2-ketoisocaproate. The inhibitor of ornithine decarboxylase D,L-alpha-difluoromethyl ornithine failed to affect the metabolism and insulinotropic action of D-glucose in pancreatic islets, and only caused a partial inhibition of the secretory response to either L-arginine or L-ornithine. The latter amino acids inhibited modestly but significantly D-glucose utilization and oxidation by pancreatic islets. These and complementary findings suggest that the secretory response to L-arginine and L-ornithine is not attributable to any major change in the overall oxidative catabolism of nutrients, but involves mainly a biophysical component, such as the depolarization of the plasma membrane by these cationic amino acids.

Animals↗

PubMed1989

Anomeric specificity of D-glucose phosphorylation by rat liver glucose-6-phosphatase.

The anomeric specificity of D-glucose phosphorylation by hepatic glucose-6-phosphatase was examined in rat liver microsomes incubated in the presence of carbamoyl phosphate. At 10 degrees C, the Km for the equilibrated hexose and phosphate donor was close to 56 mM and 11 mM, respectively. The enzymic activity, which was increased in diabetic rats, was about 40% lower in untreated than in sonicated microsomes. No anomeric difference in affinity was found in sonicated microsomes. In untreated microsomes, however, the Km for beta-D-glucose was slightly lower than that for alpha-D-glucose. The maximal velocity was higher with beta- than alpha-D-glucose in both untreated and sonicated microsomes. These data indicate that the phosphotransferase activity of glucose-6-phosphatase cannot account for the higher rate of glycolysis and glycogen synthesis found in hepatocytes exposed to alpha- rather than beta-D-glucose.

Animals↗

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release. Insulinotropic action of agmatine.

Agmatine causes a concentration-related (0.1-1.0 mM) stimulation of insulin secretion by rat pancreatic islets exposed to D-glucose (5.6-11.1 mM) but, like L-arginine, fails to affect insulin release either in the absence of glucose or at a high concentration (16.7 mM) of the hexose. Agmatine, which accumulates in islet cells, also stimulates 45Ca net uptake by the islets. The secretory response to agmatine represents a delayed process. It is enhanced by theophylline and suppressed by antimycin A or the absence of extracellular Ca2+. The insulinotropic action of agmatine is compatible with the view that endogenously formed polyamines may play a role in the secretory response of islet cells to L-arginine.

Agmatine↗

PubMed1989

Effect of a phospho-oligosaccharidic putative insulin messenger on insulin release in rats.

The phospho-oligosaccharide extracted from rat liver and supposed to act as the insulin second messenger inhibits glucose-stimulated insulin release. In the present study, this phospho-oligosaccharide was found not to affect D-[U-14C]glucose oxidation and 45Ca net uptake, but to inhibit insulin release evoked by either D-glucose or 2-ketoisocaproate in isolated rat islets. The relative extent of the latter inhibition was unaffected by either the concentration of D-glucose or the presence of dibutyryl-cyclic AMP, forskolin or glucagon in the incubation medium. At variance with the inhibitory effect of clonidine, that of the phospho-oligosaccharide was resistant to both blockade of alpha 2-adrenergic receptors or pre-treatment with the toxin of Bordetella pertussis. It is speculated, therefore, that such a phospho-oligosaccharide might interfere with a distal event in the insulin secretory sequence.

Animals↗

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release: metabolism of L-arginine and L-ornithine in tumoral islet cells.

The metabolism of L-arginine and L-ornithine was examined in tumoral islet cells of the RINm5F line and compared to the situation previously characterized in normal rat islets. The maximal velocity of arginase in cell homogenates, as well as either the production of 14C-urea or the steady-state content of 14C-labelled ornithine in intact cells exposed to L-[U-14C]arginine were about one order of magnitude lower in tumoral than normal islet cells. The activity of ornithine-glutamate transaminase was similar in both cell types, and this coincided with a comparable rate of 14C-labelled L-glutamate generation by intact cells exposed to L-[1-14C]ornithine. Despite a comparable cell content in 14C-labelled ornithine of normal and tumoral cells exposed to exogenous ornithine, the rate of di- and polyamine generation was about one order of magnitude higher in tumoral than normal islet cells, this coinciding with a much higher activity of ornithine decarboxylase in RINm5F cell than islet homogenates.

Adenoma, Islet Cell↗

PubMed1989

Inhibition by a nonmetabolized analogue of L-leucine of protein biosynthesis in tumoral pancreatic islet cells.

The effect of L-leucine, its deaminated metabolite 2-ketoisocaproate and its nonmetabolized analogue b(+/-)2-aminobicyclo[2,2,1]-heptane-2-carboxylic acid (BCH) upon protein labelling was examined in tumoral islet cells (RINm5F line) exposed to L-[4-3H]phenylalanine or L-[3-3H]serine. The interpretation of the results, in terms of changes in biosynthetic activity, was obscured by a possible interference of the tested nutrients with the uptake and further metabolism of the tracer tritiated amino acids. Nevertheless, when the cells were preincubated with the nutrient secretagogues and then incubated in the sole presence of L-[3-3H]serine, BCH, but not L-leucine or 2-ketoisocaproate, still inhibited protein labelling, this coinciding with a decrease in the ratio between TCA-precipitable and total radioactivity in the RINm5F cells. The inhibitory action of BCH was antagonized, to a limited extent, by D-glucose. It is proposed that BCH could be used as a tool to interfere with the function and growth of insulinoma cells.

Adenoma, Islet Cell↗

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release. Uptake of metabolized and nonmetabolized cationic amino acids by pancreatic islets.

In order to assess the possible role of L-arginine accumulation in islet cells as a determinant of its insulinotropic action, the uptake of L-arginine and other cationic amino acids (L-ornithine, L-homoarginine, D,L-alpha-methylornithine, D,L-alpha-difluoromethylornithine) by rat pancreatic islets was compared to the ionic and secretory responses of the islets to the same amino acids. A tight correlation was found between the net uptake of these amino acids and their capacity to stimulate 86Rb efflux, 45Ca uptake and efflux, and insulin release. In the latter respect, there was little difference between metabolized and nonmetabolized amino acids. Thus, although L-homoarginine and 4-amino-1-guanylpiperidine-4-carboxylic acid failed to act as a substrate for either arginase or amino acid aminotransferase in islet homogenates, they both stimulated 86Rb efflux, 45Ca uptake and efflux, and insulin secretion in intact islets. These findings are compatible with the view that the accumulation of these positively charged amino acids in islet cells represents an essential determinant of their secretory action. Hence, the release of insulin evoked by these amino acids could be due to depolarization of the plasma membrane with subsequent gating of voltage-sensitive Ca2+ channels and/or to some other biophysical effect, as suggested by the persistence of a sizeable secretory response to L-arginine or L-ornithine in islets perifused at a high concentrations of extracellular K+ (50 mM).

Amino Acids↗

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release: comparison with lysine-induced insulin secretion.

L-Lysine, like-L-arginine, L-ornithine, or L-homoarginine, accumulated in rat pancreatic islets and stimulated 86Rb efflux, 45Ca uptake and efflux, and insulin release in islets exposed to D-glucose (7.0 mM). The effect of L-lysine differed from that of the other cationic amino acids by such features as the absence of a threshold concentration for stimulation of insulin release, a much lesser sensitivity of the secretory response to intracellular acidification, and the stimulation of 86Rb net uptake over 60 min of incubation. This coincided with the fact that even in the absence of another exogenous nutrient, L-lysine was well oxidized, augmented NH4+ production, increased both the ATP content and ATP/ADP ratio, caused a time-related decrease in 86Rb fractional outflow, and provoked either a transient (10 mM L-lysine) or sustained (20 mM L-lysine) stimulation of insulin secretion. It is proposed, therefore, that the functional response of the pancreatic B-cell to L-lysine involves not only a biophysical mechanism similar to that responsible for the insulinotropic action of L-homoarginine, but also a significant, albeit modest, metabolic component, which reflects the capacity of L-lysine to act as a fuel in islet cells.

Animals↗

PubMed1989

Differential sensitivity to beta-cell secretagogues in cultured rat pancreatic islets exposed to human interleukin-1 beta.

The early stages of insulin-dependent diabetes mellitus are characterized by a selective inability to secrete insulin in response to glucose, coupled to a better response to nonnutrient secretagogues. The deficient glucose response may be a result of the autoimmune process directed toward the beta-cells. Interleukin-1 (IL-1) has been suggested to be one possible mediator of immunological damage of the beta-cells. In the present study we characterized the sensitivity of beta-cells to different secretagogues after human recombinant IL-1 beta (rIL-1 beta) exposure. Furthermore, experiments were performed to clarify the biochemical mechanisms behind the defective insulin response observed in these islets. Rat pancreatic islets were isolated and kept in tissue culture (medium RPMI-1640 plus 10% calf serum) for 5 days. The islets were subsequently exposed to 60 pM human recombinant IL-1 beta during 48 h in the same culture conditions as above and examined immediately after IL-1 exposure. The rIL-1 beta-treated islets showed a marked reduction of glucose-stimulated insulin release. Stimulation with arginine plus different glucose concentrations, and leucine plus glutamine partially counteracted the rIL-1 beta-induced reduction of insulin release. The activities of the glycolytic enzymes hexokinase, glucokinase, and glyceraldehyde 3-phosphate dehydrogenase, were similar in control and IL-1-exposed islets. Treatment with IL-1 also did not impair the activities of NADH+- and NADPH+-dependent glutamate dehydrogenase, glutamate-aspartate transaminase, glutamate-alanine transaminase, citrate synthase, and NAD+-linked isocitrate dehydrogenase. The oxidation of D-[6-14C]glucose and L-[U-14C]leucine were decreased by 50% in IL-1-treated islets. Furthermore, there was a significant decrease in the ratios of [2-14C]pyruvate oxidation/[1-14C]pyruvate decarboxylation and L-[U-14C]leucine oxidation/L-[1-14C]leucine decarboxylation, indicating that IL-1 decreases the proportion of generated acetyl-coenzyme-A residues undergoing oxidation. However, in the presence of IL-1 there was a significant increase in L-[U-14C]glutamate oxidation. These combined observations suggest that exposure to IL-1 induces a preferential decrease in glucose-mediated insulin release and mitochondrial glucose metabolism. This mitochondrial dysfunction seems to reflect an impairment in proximal steps of the Krebs cycle. It is conceivable that the IL-1-induced suppression and shift in islet metabolism can be an explanation for the beta-cell insensitivity to glucose observed in the early phases of human and experimental insulin-dependent diabetes mellitus.

Animals↗

PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release. Effect of 2-aminoisobutyric acid upon ornithine decarboxylase activity and insulin secretion.

Recent studies have revealed that, in rat pancreatic islets, exogenous L-arginine or L-ornithine are converted to putrescine, spermidine and spermine. The possible role of polyamine generation in the secretory response of the B-cell to the cationic amino acids was investigated in islets either preincubated for 90 min with 2-aminoisobutyric acid (AIB; 10 mM) or removed from rats injected intraperitoneally with AIB (2.4 mmol) 210 min before sacrifice. Although AIB failed to exert any direct effect upon the secretory response of the islets to either L-arginine or L-ornithine, preincubation of the islets with AIB increased both the activity of ornithine decarboxylase and, to a modest extent, the insulinotropic action of L-arginine and L-ornithine. Likewise, after treatment with AIB in vivo, the activity of ornithine decarboxylase increased tenfold or more in kidney, liver, pancreas, parotid gland and islet homogenates. The secretory response to D-glucose was little affected in islets removed from AIB-treated rats, but the insulinotropic action of L-arginine and L-ornithine were slightly increased. These findings suggest that the de novo generation of polyamines only plays a restricted role in the secretory response of islet cells to L-arginine and L-ornithine.

Amino Acids↗

PubMed1989

Activity and kinetics of liver 6-phosphofructokinase in normal and diabetic rats.

The activity and kinetics of phosphofructokinase were measured in the postmicrosomal supernatant of livers removed from either normal or streptozotocin-induced diabetic rats. The activity of the enzyme was lower in diabetic than normal rats. However, the relative magnitude of changes in reaction velocity evoked by different concentrations of D-fructose 6-phosphate, ATP, D-glucose 1,6-bisphosphate and/or D-fructose 2,6-bisphosphate were comparable in normal and diabetic rats. Such was also the case after separation of cytosolic proteins from low molecular weight metabolites by gel filtration chromatography. These findings suggest that a sustained increase in intracellular D-glucose concentration, as presumably occurring in hepatocytes of diabetic animals, does not result in any obvious alteration of the intrinsic kinetic properties of phosphofructokinase.

Animals↗

PubMed1988

Hexose metabolism in pancreatic islets. Feedback control of D-glucose oxidation by functional events.

A rise in extracellular D-glucose concentration in pancreatic islet cells causes a greater relative increase in the oxidation of pyruvate and acetyl residues than in glycolysis. A possible explanation for such an unusual situation was sought in the present study. The preferential stimulation of mitochondrial oxidative events was found to display a sigmoidal dependency on hexose concentration, and an exponential time course during prolonged exposure of the islets to a high concentration of D-glucose. The preferential stimulation of mitochondrial oxidative events was abolished in islets incubated in the presence of cycloheximide and absence of Ca2+, in which case the oxidation of D-[6-14C]glucose was more severely inhibited than that of D-[3,4-14C]glucose. Likewise, the inhibitor of protein biosynthesis and the absence of Ca2+ affected the oxidation of L-[U-14C]leucine preferentially, relative to that of L-[1-14C]leucine, in islets exposed to a high, but not a low, concentration of the amino acid. These results demonstrate that in pancreatic islets it is possible to dissociate both glycolysis from mitochondrial oxidative events and the oxidation of acetyl residues from their generation rate. Moreover, the experimental data suggest that nutrient-responsive and ATP-requiring functional processes exert a feedback control on mitochondrial respiration in this fuel-sensor organ.

Animals↗

PubMed1988

Stimulation by D-glucose of mitochondrial respiration.

D-glucose increases O2 uptake by cerebellum mitochondria. This effect is abolished by D-glucose-6-phosphate and D-mannoheptulose. It is proposed that the phosphorylation of D-glucose as catalyzed by bound hexokinase directly affects mitochondrial respiration.

Adenosine Diphosphate↗

PubMed1988

Anomeric specificity of D-glucose metabolism in human erythrocytes.

The metabolism of D-[5-3H]glucose and D-[U-14C]glucose displays anomeric specificity in human erythrocytes. At close-to-physiological hexose concentrations, pure beta-D-[5-3H]glucose is metabolized at a higher rate than the corresponding pure alpha-anomer in most, but not all, subjects. This situation represents a mirror image of that found in rat erythrocytes. The anomeric preference is modulated by the extracellular concentration of D-glucose and may differ for distinct metabolic variables. The anomeric specificity of D-glucose metabolism remains operative, whether at low or normal temperature, even in erythrocytes exposed to equilibrated D-glucose.

Blood Glucose↗

PubMed1988

Hexose metabolism in pancreatic islets. Metabolic and secretory responses to D-fructose.

D-Fructose (3.3 to 33.0 mmol/liter) caused a concentration-related increase in insulin output from rat islets exposed to D-glucose (3.3 to 7.0 mmol/liter), such an increase not being more marked in mouse islets. The fructose-induced increment in insulin release, relative to that evoked by D-glucose, was two times higher in islets exposed to D-glucose than in islets stimulated by D-mannose, 2-ketoisocaproate, or nonnutrient secretagogs. Likewise, the metabolism of D-fructose in islet cells was significantly different in the absence or presence of D-glucose. Thus, the ketose was largely channeled into the pentose phosphate pathway in glucose-deprived, but not so in glucose-stimulated, islets. In both glucose-deprived and glucose-stimulated islets, however, the magnitude of the secretory response to D-fructose was commensurate with the increase in ATP production attributable to its catabolism. These findings indicate that the metabolic fate of hexoses--and, hence, their insulinotropic capacity--is not ruled solely at the level of their phosphorylation.

Animals↗

PubMed1988

Crabtree effect in tumoral pancreatic islet cells.

The relationship between glycolysis and respiration was examined in a model of pancreatic B-cell dysfunction, namely in tumoral insulin-producing cells of the RINm5F line. A rise in D-glucose concentration from 2.8 to 16.7 mM increased the utilization of D-[5-3H]glucose and production of [14C]lactate from D-[U-14C]glucose, whereas decreasing the oxidation of either D-[U-14C]glucose or D-[6-14C]glucose. Whereas 2.8 mM D-glucose augmented O2 uptake above basal value, a further rise in D-glucose concentration to 16.7 mM decreased respiration, which remained higher, however, than basal value. Whether at low or high concentration, D-glucose exerted a pronounced sparing action upon the oxidation of endogenous nutrients in cells prelabeled with either L-[U-14C]glutamine or [14C]palmitate and, nevertheless, augmented above basal value the rate of lipogenesis, ATP/ADP content, adenylate charge, and cytosolic NADH/NAD+ and NADPH/NADP+ ratios. The generation of ATP resulting from the catabolism of either exogenous D-glucose or endogenous nutrients was not affected by the rise in hexose concentration from 2.8 to 16.7 mM. Thus, in sharp contrast with the situation found in normal islet cells, a rise in D-glucose concentration, instead of stimulating mitochondrial oxidative events, caused, through a Crabtree effect, inhibition of hexose oxidation and O2 consumption in tumoral islet cells.

Adenine Nucleotides↗

PubMed1988

Hexose metabolism in pancreatic islets stimulation by D-glucose of [2-3H]glycerol detritiation.

1. In pancreatic islets, a rise in glucose concentration is known to increase the ratio between D-[6-14C]glucose oxidation and D-[5-3H]glucose utilization. The opposite situation was found to prevail in parotid cells. 2. In rat pancreatic islets, D-glucose caused a concentration-related stimulation of 3H2O production from [2-3H]glycerol, but failed to affect 3H2O production from [1(3)-3H]glycerol or 14CO2 production from [U-14C]glycerol. At the low concentration used in most of these experiments (i.e. 1.0 mM), glycerol failed to affect D-[U-14C]glucose oxidation. 3. These findings suggest that the preferential stimulation by D-glucose of mitochondrial oxidative events in pancreatic islets represents an unusual situation in secretory cells and involves an accelerated circulation in the glycerol phosphate shuttle.

Anaerobiosis↗

PubMed1988

Underestimation of D-glucose phosphorylation as measured by 3H2O production from D-[2-3H]glucose.

In rat pancreatic islets, tumoral islet cells (RINm5F line), parotid gland, and in human erythrocytes, but not in rat hepatocytes, the production of 3H2O from D-[2-3H]glucose is 20-30% lower than from D-[5-3H]glucose. This coincides with the production of tritiated lactic acid from D-[2-3H]glucose and may be attributable to an intramolecular hydrogen transfer in the phosphoglucoisomerase reaction. It is concluded that the production of 3H2O from D-[2-3H]glucose is not a reliable tool to assess the total rate of hexose phosphorylation.

Animals↗