PubMed Health⌕ Search

Biomedical subjects

B Hellman

Publications and source records attributed to B Hellman.

At least 73 records · Page 4Linked to original sources
PubMed1992

Stimulation of insulin release by isosmolar addition of permeant molecules.

Pancreatic beta-cells are known to respond to hyposmolar stress by releasing insulin. It was evident from perifusion studies using islet cells from ob/ob-mice mixed with polyacrylamide beads that a similar type of secretory response can be obtained by isosmolar addition of 10-25 mM of the rapidly penetrating urea molecule. There was no effect with hyperosmolar addition of urea. The urea-induced insulin release differed from the ordinary stimulation of secretion in not disappearing but being more pronounced after previous heating to 45 degrees C or removal of extracellular Ca2+. Isosmolar urea was exceptional as an insulin secretagogue in being effective also in the presence of the alpha 2-adrenergic agonist clonidine or when lowering the temperature to 24 degrees C. Further support for the idea that isosmolar addition of rapidly penetrating molecules induces insulin release was obtained by testing non-metabolizable glucose analogues. Whereas 25 mM 3-O-methyl-D-glucose doubled the secretory rate within 4 min, the non-permeant L-glucose had only a slight initial action. When not compensating for the alterations of the medium osmolarity 3-O-methyl-D-glucose was without effect. Although expansion of beta-cells cannot explain the existence of a pronounced initial secretory response to D-glucose it may under certain conditions contribute to the stimulatory effects of the sugar.

3-O-Methylglucose↗
PubMed1992

Glucose sensing of individual pancreatic beta-cells involves transitions between steady-state and oscillatory cytoplasmic Ca2+.

Glucose stimulation of individual pancreatic beta-cells is associated with a rise of the cytoplasmic Ca2+ concentration ([Ca2+]i) manifested either as large amplitude oscillations (0.2-0.5/min) or as a sustained increase. Determinants for the transitions between the basal and the two stimulated states have now been studied using dual-wavelength fluorometric measurements on individual ob/ob mouse beta-cells loaded with the Ca2+ indicator Fura-2. The transition from the basal state to large amplitude oscillations was induced by raising the glucose concentration to 7 mM or above. The frequencies and shapes of the [Ca2+]i cycles remained largely unaffected when raising glucose as high as 40 mM. However, in some cells the oscillatory pattern was transformed into a sustained increase of [Ca2+]i at high glucose concentrations. Although the peak values for the oscillations exceeded the steady-state increase, the time average [Ca2+]i was higher during the latter phase. Both types of glucose-induced transitions were facilitated by the presence of 1-100 nM glucagon. Protein kinase C activation by 10 nM of the phorbol ester TPA resulted in a transformation of the glucose-induced oscillations into a sustained increase of [Ca2+]i but the levels reached were considerably lower than obtained with glucose alone. It is concluded that the glucose sensing of the individual beta-cell is based on sudden transitions between steady-state and oscillating cytoplasmic Ca2+. It is these transitions rather than alterations of the oscillatory characteristics which determine the average [Ca2+]i regulating insulin release.

Animals↗
PubMed1991

Carbachol has opposite effects to glucose in raising the sodium content of pancreatic islets.

Integrating flame photometry was used for measuring sodium in single pancreatic islets from ob/ob mice. Exposure to 100 microM carbachol resulted in a 25-40% increase in sodium without any effect on potassium during incubation with 0-5 mM glucose in media deficient or not in Ca2+. This action of carbachol was abolished by 10 microM atropine or by raising the glucose concentration to 20 mM. A minor increase of the steady state content of sodium occurred in the presence of 200 microM ATP or 10 nM tetradecanoylphorbol 13-acetate (TPA). Carbachol differed from TPA in markedly stimulating sodium accumulation after ouabain inhibition of the Na/K pump. The results indicate that muscarinic receptor activation has opposite effects to glucose in inducing a rise of the islet content of sodium. It is suggested that the cholinergic control of the endocrine pancreas involves entry of Na+ in addition to the Na+ entry mediated by protein kinase C activation of Na+/H+ countertransport.

Adenosine Triphosphate↗
PubMed1991

Cyclic AMP as a determinant for glucose induction of fast Ca2+ oscillations in isolated pancreatic beta-cells.

The effect of glucose on the cytoplasmic Ca2+ concentration ([Ca2+]i) of pancreatic beta-cells from ob/ob-mice was examined by dual wavelength recordings of the 340/380 nm fluorescence excitation ratio of fura-2. Single beta-cells responded to 11-20 mM glucose with an initial lowering of [Ca2+]i, followed by an increase usually manifested as large amplitude oscillations (300-500 nm) with a frequency of 0.2-0.5/min (a-type). Particularly in freshly isolated beta-cells, there were also superimposed fast oscillations with frequencies of 2-8/min amplitudes in the 70-250 nM range (b-type) and sometimes pronounced [Ca2+]i transients exceeding 250 nM with durations below 10 s (c-type). After addition of 1-100 nM glucagon or 1 mM of the dibutyryl or 8-bromo derivatives of cyclic AMP, glucose generated numerous b-type oscillations superimposed on those of the a-type or on an elevated steady-state level. The duration of the b-type oscillations increased slightly when glucose was raised from 11 to 16 mM. The c-type transients probably represent a separate reaction predominantly seen when raising cyclic AMP much above its normal concentration. It is concluded that glucose can induce fast oscillations of [Ca2+]i also in isolated beta-cells, especially when measures are taken to increase their cyclic AMP content.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1991

Intracellular ATP mimics GTP-gamma-S in generating Ca2+ oscillations in pancreatic beta-cells.

Intracellular free calcium ([Ca2+]i) was measured in individual pancreatic beta-cells from mice using dual emission microfluorometry and the indicator Indo-1 applied by a patch clamp pipette. GTP-gamma-S (100 microM) injected together with 0.3 or 3 mM ATP evoked repetitive [Ca2+]i transients with a frequency of about 1 per min in beta-cells kept at a membrane potential of -70 mV. The oscillatory pattern was unaffected by the Ca2+ channel blocker verapamil (50 microM). When omitting GTP-gamma-S from the pipette medium it became evident that 3 mM ATP alone can induce oscillations. The results provide additional evidence for an important role of ATP in the ionic control of insulin release, indicating that such regulation may also involve activation of G-proteins.

Adenosine Triphosphate↗
PubMed1991

Glucose stimulation of ouabain-resistant efflux of Na+ from rat pancreatic islets.

1. Integrating flame photometry was employed for measuring the mobilization of sodium from rat pancreatic islets after substitution of extracellular Na+ by N-methylglucamine. 2. Glucose accelerated the initial loss of sodium both in the absence and presence of ouabain (1 mM). In the latter case the effect was maximal at 5 mM of the sugar. 3. Amiloride (0.1 mM), an inhibitor of Na(+)-H+ exchange, prevented the effect of glucose on the ouabain-resistant Na+ efflux, increasing the rate of outward transport in the absence of the sugar. 4. Extracellular K+ and arginine (10 mM) mimicked the action of glucose in promoting a ouabain-resistant mobilization of sodium. 5. Whereas the hypoglycaemic sulphonylurea tolbutamide (100 microM) did not modify the outward transport of Na+, the ouabain-resistant component of this process was partially suppressed after bumetanide (100 microM) inhibition of the chloride-dependent co-transport of Na+ and K+. 6. It is suggested that the glucose-induced lowering of the steady-state content of islet sodium involves an increased outward transport mediated at least in part by mechanisms other than stimulation of the Na(+)-K+ pump.

Animals↗
PubMed1991

Glucose has regulatory effects on insulin release also in the virtual absence of extracellular Ca2+.

The glucose effect on insulin release in a Ca(2+)-deficient medium was analyzed in perifusion experiments with aggregates of cells prepared by dispersal of the beta-cell-rich pancreatic islets of ob/ob-mice. Hyperosmolar additions of 20 mM D-glucose or its poorly metabolized transport analogue 3-0-methyl-D-glucose resulted in 50% suppression of the secretory rate. However, after isosmolar additions of the sugars, replacing non-penetrating sucrose, there was a stimulation of insulin release. Whereas D-glucose was less effective than 3-O-methyl-D-glucose in stimulating insulin release after isosmolar addition, the opposite was found for the enhanced secretory response obtained when the sugars were excluded from the perifusion medium. The studies indicate that D-glucose has regulatory actions on insulin release also in the virtual absence of extracellular Ca2+. This effect is not only due to osmolar influences but involves also direct suppression of the secretory activity probably mediated by the metabolism of the sugar.

3-O-Methylglucose↗
PubMed1990

Regulatory volume decrease of pancreatic beta-cells involving activation of tetraethylammonium-sensitive K+ conductance.

Beta-cell-rich pancreatic islets from ob/ob-mice were used for evaluating the early effects of hypotonic stress. The beta-cells responded to an abrupt lowering of the osmotic pressure by 102 mOsm with both a transient stimulation of insulin release (peak value 25 times above basal) and a loss of potassium without major effects on sodium. The secretory response was obtained also in the presence of 100 microM quinine or 20 mM tetraethylammonium+. The loss of potassium was not affected by 20 mM glucose or 10 microM bumetanide, but became less apparent in the presence of 100 microM quinine and disappeared when the islets were exposed to 20 mM tetraethylammonium+. Amiloride and high concentrations of the hypoglycemic sulfonylureas tolbutamide and glibenclamide had only a slight suppressive action on potassium mobilization. Patch clamp analyses revealed an increased frequency of small channel openings after exposure to the hypotonic medium. It is concluded that the pancreatic beta-cells have the ability for a regulatory volume decrease involving activation of tetraethylammonium-sensitive K+ conductance. The stimulation of insulin release obtained by lowering the osmotic pressure seems to be related to the entry of water rather than to the ion movements responsible for the readjustment of the beta-cell volume.

Animals↗
PubMed1990

Inhibitory effects of benomyl and carbendazim on the [3H]thymidine incorporation in various organs of the mouse--evidence for a more pronounced action of benomyl.

The benzimidazole fungicides benomyl and carbendazim were compared with regard to effects on [3H]thymidine incorporation in various organs of male mice given the compounds orally at various time intervals before sacrifice. Since carbendazim is a major metabolite of benomyl, it is generally assumed that the fungicidal action and toxicity of these compounds are due to the action of carbendazim. However, whereas benomyl inhibited the [3H]thymidine incorporation into thymus, spleen, liver, kidney and testis, an equimolar amount (3.4 mmol/kg body wt) of carbendazim induced a similar effect only in testis.

Animals↗
PubMed1990

Inhibitory action of benzimidazole fungicides on the in vivo incorporation of [3H]thymidine in various organs of the mouse.

The effect of benomyl on the DNA turnover in various organs of the mouse was evaluated by measuring the incorporation of [3H]thymidine 24 hr after oral administration of different doses (1.3, 2.55 and 5.1 nmol/kg body weight) of benomyl. In the thymus, spleen and testis there was a clear relationship between dose and effect, the no-observed-effect level being 1.3 mmol/kg body weight. However, in the liver and kidney there was no obvious relationship between dose and effect, the [3H]thymidine incorporation being inhibited even at the lowest dose. Equimolar amounts of the closely related fungicide carbendazim inhibited the [3H]thymidine incorporation only in the testis. The observed differences between the two compounds was not a result of different absorption rates. Whole-body autoradiography indicated a rapid absorption and a similar distribution pattern for [phenyl(U)-14C)benomyl and [phenyl(U)-14C]carbendazim. Apart from an accumulation in the retina, liver and kidney, most other organs were almost devoid of [14C]benomyl- and [14C]carbendazim-associated radioactivity.

Animals↗
PubMed1990

Disappearance of glucose-induced oscillations of cytoplasmic Ca2+ in pancreatic beta-cells exposed to streptozotocin or alloxan.

Dual wavelength microfluorometry and the indicator fura-2 were employed for measuring cytoplasmic Ca2+ (Ca2+i) in individual pancreatic beta-cells isolated from ob/ob-mice. In most beta-cells, a rise of external glucose from 3 to 20 mM resulted in large amplitude oscillations in Ca2+i, superimposed on a basal level of 60-90 nM. The diabetogenic agents streptozotocin and alloxan (1-4.4 mM) rapidly abolished the glucose-induced oscillations of Ca2+i. The presence of a high glucose concentration during the exposure to the drugs counteracted the action of alloxan but not that of streptozotocin. Perturbation of the cyclic variations of Ca2+i by streptozotocin did not interfere with a glucose-induced increase of the ion in mildly affected beta-cells. The most advanced lesions obtained with the exposure to the diabetogenic agents were manifested as uncontrolled and sustained increases of Ca2+i. Although disrupting the intracellular Ca2+ homeostasis by separate mechanisms, streptozotocin and alloxan may finally kill the beta-cells by activating a common suicidal process due to an excessive rise of Ca2+i.

Alloxan↗
PubMed1990

The cytoplasmic Ca2+ response to glucose as an indicator of impairment of the pancreatic beta-cell function.

The effects of glucose on the cytoplasmic Ca2+ concentration (Ca2+i) regulating insulin release were investigated using pancreatic beta-cells representative for the normal and diabetic situations. Increase of the glucose concentration resulted in a slight lowering of Ca2+i followed by a rise, often manifested as high amplitude oscillations. The Ca2+i-lowering component in the glucose action associated with suppression of insulin release became particularly prominent when the beta-cells were already depolarized by tolbutamide. Glucose-induced inhibition of insulin release was observed also in experiments with rats made diabetic with streptozotocin or alloxan. Other studies indicated lowering of plasma insulin after intravenous glucose administration in patients with insulin- and noninsulin-dependent diabetes mellitus. Brief exposure of beta-cells to 2.2 mmol l-1 streptozotocin resulted in impairment of the response to glucose, manifested as disappearance of the cyclic variation of Ca2+i. The results indicate that glucose-induced depolarisation is a vulnerable process, the disturbance of which may contribute to insulin secretory defects in diabetes mellitus.

Animals↗
PubMed1990

Effect of o,p'-DDD on the in vivo incorporation of 3H-thymidine into DNA: evidence for induced cell proliferation in the mouse lung.

The effects of o,p'-DDD on the DNA synthesis in the C57Bl mouse lung and liver were studied. As determined by 3H-thymidine incorporation into DNA, a selective increase in the lung DNA synthesis (+59%) was observed 2 days after a single intraperitoneal injection of 100 mg/kg o,p'-DDD. Microautoradiography showed that the incorporated 3H-thymidine was confined to a restricted number of heavily labelled cells, presumably proliferating type II cells. At the most, a 9 times higher rate of cell proliferation was observed in the lung 4 days after an intraperitoneal injection of 500 mg/kg o,p'-DDD. Using mouse lung or liver S-9 as activating system, no mutagenic activity of o,p'-DDD was detected in the Ames test. The induced cell proliferation may indicate a tissue-selective promoter activity of o,p'-DDD in the mouse lung.

Animals↗
PubMed1990

Sulfonylurea mimics the effect of glucose in inducing large amplitude oscillations of cytoplasmic Ca2+ in pancreatic beta-cells.

The effects of the insulin-releasing sulfonylurea tolbutamide on the cytoplasmic Ca2+ concentration [( Ca2+]i) in individual pancreatic beta-cells or suspensions of beta-cells were analyzed using the probe fura-2 and dual-wavelength fluorometry. Subsequent additions of 1, 10, and 100 microM tolbutamide induced a graded response, ranging from a single [Ca2+]i peak to a sustained increase. These effects depended on the presence of extracellular Ca2+ and were reversed by the hyperglycemic sulfonamide diazoxide. The responses were diminished in the presence of albumin and varied considerably between different cells. Sometimes tolbutamide triggered slow large amplitude oscillations in [Ca2+]i similar to those induced by glucose. The increase in [Ca2+]i during each tolbutamide-induced oscillation was often more rapid than for glucose-induced oscillations. Oscillations or steady state increases in [Ca2+]i induced by glucose were little influenced by tolbutamide. However, subthreshold concentrations of glucose could reactivate [Ca2+]i response to tolbutamide that had declined. Although in several ways the abilities of glucose and tolbutamide to raise [Ca2+]i were similar, the sulfonylurea lacked a [Ca2+]i-lowering component. The latter effect of glucose was so pronounced that an increase of its concentration from 3 to 20 mM caused temporary lowering of [Ca2+]i to the basal level, even during tolbutamide stimulation. The results indicate that closure of the ATP-sensitive K(+)-channels is important for the large amplitude oscillations of [Ca2+]i the appearance of which reflects the balance between entry of Ca2+ through the voltage-dependent channels and its removal from the cytoplasm.

Animals↗
PubMed1989

Regulation of pH in individual pancreatic beta-cells as evaluated by fluorescence ratio microscopy.

Pancreatic beta-cells are known to maintain intracellular pH (pHi) at a value well above that predicted from the electrochemical gradient. The mechanisms for the active extrusion of protons were examined by continuously monitoring pHi in individual beta-cells from ob/ob mice using the fluorescent indicator 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). In a medium nominally devoid of bicarbonate, the steady-state pHi was 6.82 +/- 0.02 and the intracellular buffering capacity was equivalent to 79 +/- 3 mM/pH unit. pHi remained unaffected after raising the glucose concentration from 3 to 20 mM, it was lowered when depolarizing the beta-cells with tolbutamide and it increased in the presence of carbachol. After removal of Na+ there was a significant drop of pHi and blockage of the pHi recovery following acid loading with the NH4+ prepulse technique. Whereas addition of amiloride had a similar, but less pronounced effect, omission of Cl- resulted in moderate alkalinisation. After switching to a medium containing bicarbonate, minor acidification was followed by adjustment of pHi to a steady state higher than the initial one. The results indicate that the acid load arising from glucose metabolism in the beta-cells is effectively buffered and the protons extruded both by Na+-H+ and Cl- -HCO3- exchangers.

Amiloride↗
PubMed1989

Free and bound sodium in pancreatic beta-cells exposed to glucose and tolbutamide.

The effects of glucose and tolbutamide on the sodium handling of the pancreatic beta-cells were evaluated by measuring the total sodium content in intact islets from ob/ob-mice by integrating flame photometry and the free ion in individual beta-cells by dual wavelength fluorometry. Whereas increasing the glucose concentration from 3 to 20 mM resulted in a lowering of sodium, the addition of 100 microM tolbutamide caused a rise. The above-mentioned effects were most marked (about 50%) for the physiologically significant free sodium. The data indicate a more important role for Na+ in the regulation of insulin release than so far acknowledged. Increase of Na+ may contribute to the secretory response to hypoglycemic sulfonylureas by providing an additional rise of cytoplasmic Ca2+.

Animals↗
PubMed1989

Effects of depolarizing agents on the sodium content of rat pancreatic islets.

Rat pancreatic islets were used for studying the effects of depolarization on their sodium content. The islet sodium was markedly affected by small variations of extracellular K+. As with increased K+, the presence of low concentrations of glucose (5 mM) and arginine (2 mM) decreased the sodium content. The latter substances did not lower the sodium concentration below the value obtained by depolarization with excessive K+, nor was it possible to obtain a further decrease when 10 mM arginine was combined with 5 mM glucose. The sodium content was also reduced in the presence of 10 mM L-leucine, 10 mM 2-ketoisocaproate and 0.1 mM Ba2+. Tolbutamide differed from the other depolarizing agents in that it increased the sodium concentration, an effect manifested also in the presence of excessive K+. The observation that depolarizing agents other than sulfonylureas do not increase but actually reduce sodium implies that islet cells are exceptional among electrically excitable cells. The observed reduction of sodium may reflect activation of a voltage-sensitive carrier mechanism for outward transport of Na+.

Animals↗