Regulation of energy metabolism in pancreatic islets by glucose and tolbutamide

Summary The kinetics of insulin secretion and oxygen uptake in response to D-glucose and tolbutamide were compared in mouse pancreatic islets. In addition, the role of decreased ATP as a driving force for secretagogue-induced oxygen consumption was examined. D-glucose (10–30 mmol/1) triggered a biphasic insulin release which always coincided with a monophasic increase in islet oxygen uptake. In the presence of D-glucose (5–30 mmol/1), tolbutamide (3–500 g,mol/1) consistently elicited an initial peak of insulin secretion which was followed by a continued decline. Tolbutamide-induced secretory profiles were accompanied by similar respiratory profiles. Oxygen consumption per ng of insulin released during the test phase was higher after elevation of the glucose concentration than after addition of tolbutamide. In conjunction with 5 or 10 mmol/l D-glucose, but not with 15 or 30 mmol/1 D-glucose, tolbutamide (30–100 mol/1) lowered islet ATP content significantly (p < 0.02). Phosphocreatine was not found in isolated islets, although they contained substantial creatine kinase activity. It is concluded that the driving force for Tobutamide-induced oxygen uptake is a decrease in the phosphorylation potential caused by the work load imposed by stimulation of the secretion process. However, a major proportion of the respiratory response to glucose also results from enhancement of biosynthesis.     

Oscillations in cytoplasmic free calcium concentration in human pancreatic islets from subjects with normal and impaired glucose tolerance

Summary Plasma insulin levels in healthy subjects oscillate and non-insulin-dependent diabetic patients display an irregular pattern of such oscillations. Since an increase in cytoplasmic free Ca²⁺ concentration ([Ca²⁺]_i) in the pancreatic beta cell is the major stimulus for insulin release, this study was undertaken to investigate the dynamics of electrical activity, [Ca²⁺]_i-changes and insulin release, in stimulated islets from subjects of varying glucose tolerance. In four patients it was possible to investigate more than one of these three parameters. Stimulation of pancreatic islets with glucose and tolbutamide sometimes resulted in the appearance of oscillations in [Ca²⁺]_i, lasting 2–3 min. Such oscillations were observed even in some islets from patients with impaired glucose tolerance. In one islet from a diabetic patient there was no response to glucose, whereas that islet displayed [Ca²⁺]_i-oscillations in response to tolbutamide, suggesting that sulphonylurea treatment can mimic the complex pattern of glucose-induced [Ca²⁺]_i-oscillations. We also, for the first time, made patch-clamp recordings of membrane currents in beta-cells in situ in the islet. Stimulation with glucose and tolbutamide resulted in depolarization and appearance of action potentials. The islet preparations responded to stimulation with a number of different secretagogues with release of insulin. The present study shows that human islets can respond to stimulation with glucose and sulphonylurea with oscillations in [Ca²⁺]_i, which is the signal probably underlying the oscillations in plasma insulin levels observed in healthy subjects. Interestingly, even subjects with impaired glucose tolerance had islets that responded with oscillations in [Ca²⁺]_i upon glucose stimulation, although it is not known to what extent the response of these islets was representative of most islets in these patients.Abbreviations [Ca²⁺]_i Cytoplasmic free Ca²⁺ - NIDDM non-insulin-dependent diabetes mellitus - DMSO dimethylsulphoxide - PC pancreatic cancer     

Effect of complement depletion on O2 supply and consumption in ischemic dog myocardium

Summary The purpose of this study was to determine whether depletion of serum complement can decrease the severity of an ischemic episode by improving regional O₂ supply and conumption parameters in the ischemic region of the heart. Fourteen anesthetized dogs with serum complement intact or depleted (100 U/kg cobra venom factor given 8 hrs before) were subjected to left anterior descending coronary artery (LAD) occlusion for 6 hrs. Myocardial blood flows were determined before and 6 hrs after LAD occlusion using radioactive microspheres. Regional arterial and venous O₂ saturations were determined using microspectrophotometry. In control animals, flow decreased from 122±42 to 13±14 ml/min/100 g (mean±SD) in the occluded LAD region. With complement depletion, LAD occlusion resulted in a flow reduction in the ischemic region (38±29 ml/min/100 g), but to a lesser degree than seen in the same region in control animals, especially in the subendocardium. O₂ consumption was decreased in the ischemic region of both treatment groups, though O₂ consumption was higher in this region in complement depleted animals compared to the values in control animals. The O₂ supply/consumption ratio was decreased similarly in the ischemic region of control and complement depleted groups. Thus, with complement depletion, flow to the ischemic zone was improved but this region was still flow restricted. The flow increase during complement depletion was sufficient to allow an increased O₂, utilization in the ischemic region.     

Glucagon-like peptide-1 but not glucagon-like peptide-2 stimulates insulin release from isolated rat pancreatic islets

Summary Glucagon-like peptide-1 and glucagon-like peptide-2 are encoded by the m-RNA of pancreatic preproglucagon. They show high conservation in different species and substantial sequence homology to glucagon. Because no definite biological activity of these peptides has been reported, we investigated the effect of synthetic C-terminally amidated glucagon-like peptide-1 [1–36] and synthetic human glucagon-like peptide-2 [1–34] with a free C-terminus on insulin release from isolated precultured rat pancreatic islets in the presence of glucose. Glucagon-like peptide-1 stimulates insulin release at 10 and 16.7 mmol/l glucose in a dose-dependent manner. Significant stimulation starts at 2.5 nmol/l in the presence of 10 mmol/l glucose and near maximal release is observed at 250 nmol/l, with approximately 100% increase over basal at both glucose concentrations. The peptide reaches 63% of the maximal stimulatory effect of glucagon. No stimulation occurs in the presence of 2.8 mmol/l glucose. Glucagon-like peptide-2 has no effect on insulin secretion at any glucose concentration tested. It is concluded that glucagon-like peptide-1, in contrast to glucagon-like peptide-2, exhibits a glucose-dependent insulinotropic action on isolated rat pancreatic islets similar to that of glucagon and gastric inhibitory polypeptide.     

The pancreas and oxygen consumption (Part 2) Steroids do not modify resting pancreatic oxygen consumption in dogs: A preliminary report

In order to evaluate the effect of methylprednisolone sodium succinate (MPSS) on the alteration of pancreatic oxygen consumption (VO2) in hypovolemic shock, MPSS was administered to four normal canines and three hypovolemic animals. All were treated according to the protocol used in the initial report. (The Pancreas and Oxygen Consumption 1: Pancreatic Oxygen Consumption in Normo- and Hypovolemic Dogs.) All seven underwent a splenectomy at the beginning of the experiment. Pancreatic VO₂, obtained by adding up VO₂ for the head (minus the uncinate process) and tail of the pancreas, was equal to the product of regional blood flow,Q, determined electromagnetically on the gastroduodenal (GDA) and splenic (SA) arteries, times O₂ extraction, (a-v)O₂; O₂ content (in mL%) was measured in the femoral artery (RFA), in the splenic (SV) and superior pancreaticoduodenal (SPDV) veins. Similar determinations were carried out on the right hind limb that served as a control. Recordings were made for 4 h in both groups, the first hour determinations (five in all) serving as reference values. Methylprednisolone did not appear to alter pancreatic VO₂, which showed a significant increasing trend from + 77% 1 h after MPSS had been given, to + 98% 3 h later (vs + 56 and +92%, respectively, in the control group). As in the control group, these increases were owing to augmented O₂ extraction by the pancreas. No significant change was noted between the head and tail of the pancreas. In the hind limb, VO₂ increased significantly the first 2 h and differed from control VO₂ at the end of the first hour only.     

Fentanyl inhibits glucose-stimulated insulin release from β-cells in rat pancreatic islets

AIM： TO explore the effects of fentanyl on insulin release from freshly isolated rat pancreatic islets in static culture. METHODS： Islets were isolated from the pancreas of mature Sprague Dawley rats by common bile duct intraductal collagenase V digestion and were purified by discontinuous Ficoll density gradient centrifugation. The islets were divided into four groups according to the fentanyl concentration： control group （0 ng/mL）, group I （0.3 ng/mL）, group I （3.0 ng/mL）, and group III （30 ng/mL）. In each group, the islets were co-cultured for 48 h with drugs under static conditions with fentanyl alone, fentanyl ＋ 0.1 μg/mL naloxone or fentanyl ＋ 1.0 μg/mL naloxone. Cell viability was assessed by the MTT assay. Insulin release in response to low and high concentrations （2.8 mmol/L and 16.7 mmol/L, respectively） of glucose was investigated and electron microscopy morphological assessment was performed. RESULTS： Low- and high-glucose-stimulated insulin release in the control group was significantly higher than in groups I and II （62.33 ± 9.67 μIU vs 47.75 ± 8.47 μIU, 39.67 ± 6.18 μIU and 125.5 ± 22.04 μIU vs 96.17 ± 14.17 μIU, 75.17 ± 13.57 μIU, respectively, P 〈 0.01） and was lowest in group III （P 〈 0.01）. After adding 1 μg/mL naloxone, insulin release in groups II and II was not different from the control group. Electron microscopy studies showed that the islets were damaged by 30 ng/ml fentanyl. CONCLUSION： Fentanyl inhibited glucose-stimulated insulin release from rat islets, which could be prevented by naloxone. Higher concentrations of fentanyl significantly damaged β-cells of rat islets.     

The role of reduced glucose transporter content and glucose metabolism in the immature secretory responses of fetal rat pancreatic islets

Summary Isolated fetal islets show an immature or poor secretory response to nutrient secretagogues which may result from impaired mitochondrial oxidative processes. Insulin secretion, glucose metabolism and detection of metabolic enzymes by radiolabelling and immunoprecipitation were compared in islets isolated from neonatal (aged 5 days) and fetal rats (at 20 days gestation). The insulin secretory dynamics of fetal islets were abnormal in response to stimulation by glucose (10 mmol/l); a rapid release of insulin reaching a maximum 6 min after stimulation was observed with no rising second phase release. However, when the data were expressed as percentage of islet insulin content released, fetal islets released significantly more insulin than neonatal islets in response to glucose (4.86±0.45 % vs 1.81±0.62 %, p <0.01) or 100 nmol/l glibenclamide (2.49±0.17 % vs 0.25±0.06 %, p < 0.001). Fetal islets however, failed to release insulin in response to stimulation by glyceraldehyde (10 mmol/l) unlike neonatal islets. Both glucose utilisation (as measured by the formation of [³H] H₂O from 5-[³H] glucose) and glucose oxidation (as measured by the formation of [¹⁴C] CO₂ from U-[¹⁴C] glucose) did not increase significantly in response to increasing the medium glucose concentration to 10 mmol/l whereas in neonatal islets, glucose utilisation and glucose oxidation were significantly increased 2.5- and 2.7-fold, respectively. When islets were incubated with both radiolabelled glucoses simultaneously, the rate of glucose oxidation was shown to be directly proportional to the rate of glucose utilisation. The relationship between glucose utilisation and glucose oxidation was similar in fetal and neonatal islets. Finally, in experiments to detect and semiquantify metabolic enzymes, the level of GLUT-2 transporter protein was significantly reduced by 50 % (p <0.02) whereas the levels of pyruvate dehydrogenase peptides were similar in fetal and neonatal islets. In conclusion, these data do not support the hypothesis that abnormal mitochondrial oxidation is responsible for the immature secretory responses to nutrient secretagogues found in fetal islets but rather that step(s) earlier in the glycolytic pathway are important for development of normal secretory function. [Diabetologia (1994) 37: 134–140] Received: 1 June 1993 and in revised form: 16 August 1993     

Drastic Dependence of the pH Sensitivity of Fe2O3-Bi2O3-B2O3 Hydrophobic Glasses with Composition

Fe₂O₃-Bi₂O₃-B₂O₃ (FeBiB) glasses were developed as novel pH responsive hydrophobic glasses. The influence of the glass composition on the pH sensitivity of FeBiB glasses was investigated. The pH sensitivity drastically decreased with decreasing B₂O₃ content. A moderate amount of Fe₂O₃ and a small amount of B₂O₃ respectively produces bulk electronic conduction and a pH response on glass surfaces. Because the remaining components of the glass can be selected freely, this discovery could prove very useful in developing novel pH glass electrodes that are self-cleaning and resist fouling.     

Permissive effect of glucose on the glucagon-induced accumulation of cAMP in isolated rat pancreatic islets

Summary Glucose stimulation increased the cAMP content of collagenase-isolated rat pancreatic islets fourfold above baseline values. The elevation was transient, lasting about 5 min, and was dose-dependent. Insulin release continued at a constant rate throughout the incubation. — Glucagon, in the absence of glucose, increased cAMP for about 1 min, but only slightly, and had no effect on insulin release. In the presence of glucose, however, glucagon enhanced islet cAMP content 15-fold and increased the release of insulin. Glucagon was most effective at high glucose concentrations (16.6 and 25 mM). — This indicates that glucagon is critically dependent on the presence of glucose in order to increase the islet cAMP content and to stimulate insulin release. The inability of glucagon to generate sufficient cAMP in the absence of glucose might be one of the reasons why the hormone is a potentiator rather than an initiator of insulin release.     

Phosphoenolpyruvate in rat pancreatic islets: A possible intracellular trigger of insulin release?

Summary The content of phosphoenolpyruvate (PEP) has been measured in isolated rat islets of Langerhans incubated in vitro. Islet PEP was higher in islets incubated with 16.7 mmol/l glucose than in islets incubated with zero or 2.8 mmol/l glucose. Islet PEP content was also increased in islets incubated with 5 mmol/l D-glyceraldehyde. Mannoheptulose abolished the glucose-induced rise in PEP content but not that elicited by D-glyceraldehyde. These results are consistent with a role for PEP as an intracellular mediator of glucose- and glyceraldehyde-induced insulin release. The kinetics of pyruvate kinase in extracts of rat islets were studied. The maximal extractable activity was considerably higher than known rates of glycolytic flux. The Km values were found to be 0.16 mmol/l for PEP and 0.5 mmol/l for ADP. The control of islet PEP content and the possible role of PEP in insulin release are discussed.     

Atomic Layer Deposition of Ultrathin La2O3/Al2O3 Nanolaminates on MoS2 with Ultraviolet Ozone Treatment

Due to the chemically inert surface of MoS₂, uniform deposition of ultrathin high-κ dielectric using atomic layer deposition (ALD) is difficult. However, this is crucial for the fabrication of field-effect transistors (FETs). In this work, the atomic layer deposition growth of sub-5 nm La₂O₃/Al₂O₃ nanolaminates on MoS₂ using different oxidants (H₂O and O₃) was investigated. To improve the deposition, the effects of ultraviolet ozone treatment on MoS₂ surface are also evaluated. It is found that the physical properties and electrical characteristics of La₂O₃/Al₂O₃ nanolaminates change greatly for different oxidants and treatment processes. These changes are found to be associated with the residual of metal carbide caused by the insufficient interface reactions. Ultraviolet ozone pretreatment can substantially improve the initial growth of sub-5 nm H₂O-based or O₃-based La₂O₃/Al₂O₃ nanolaminates, resulting in a reduction of residual metal carbide. All results indicate that O₃-based La₂O₃/Al₂O₃ nanolaminates on MoS₂ with ultraviolet ozone treatment yielded good electrical performance with low leakage current and no leakage dot, revealing a straightforward approach for realizing sub-5 nm uniform La₂O₃/Al₂O₃ nanolaminates on MoS₂.     

Machine learning–based inverse design for electrochemically controlled microscopic gradients of O2 and H2O2

A fundamental understanding of extracellular microenvironments of O₂ and reactive oxygen species (ROS) such as H₂O₂, ubiquitous in microbiology, demands high-throughput methods of mimicking, controlling, and perturbing gradients of O₂ and H₂O₂ at microscopic scale with high spatiotemporal precision. However, there is a paucity of high-throughput strategies of microenvironment design, and it remains challenging to achieve O₂ and H₂O₂ heterogeneities with microbiologically desirable spatiotemporal resolutions. Here, we report the inverse design, based on machine learning (ML), of electrochemically generated microscopic O₂ and H₂O₂ profiles relevant for microbiology. Microwire arrays with suitably designed electrochemical catalysts enable the independent control of O₂ and H₂O₂ profiles with spatial resolution of ∼10¹ μm and temporal resolution of ∼10° s. Neural networks aided by data augmentation inversely design the experimental conditions needed for targeted O₂ and H₂O₂ microenvironments while being two orders of magnitude faster than experimental explorations. Interfacing ML-based inverse design with electrochemically controlled concentration heterogeneity creates a viable fast-response platform toward better understanding the extracellular space with desirable spatiotemporal control.

Ubiquitous spatiotemporal heterogeneity of natural environments fosters the diverse and fascinating biology that our world embraces, and motivates researchers to mimic natural environments with high spatiotemporal resolution (1–5). Given their close relevance in biochemical metabolisms, dioxygen (O₂) and hydrogen peroxide (H₂O₂) as a surrogate of reactive oxygen species (ROS) are two ubiquitous biologically relevant species in extracellular medium (1, 6). Their extracellular spatial and temporal distributions, particularly at the microscopic scale ranging from 1 μm to 100 μm (7–11), are critical for signal transduction, protein expression, biochemical redox balance, and regulation for cellular metabolism with extensive ecological, environmental, and biomedical implications (Fig. 1A) (1, 3, 8–13). A programmable creation of the spatiotemporal concentration profiles of O₂ and H₂O₂ offers the freedom to mimic, control, and perturb the microenvironments of O₂ and H₂O₂ and hence advance our understanding in microbiology.Open in a separate windowFig. 1.AI-based inverse design of electrochemically generated O₂ and H₂O₂ heterogeneities. (A) The ubiquitous spatiotemporal heterogeneities of O₂ and H₂O₂ in microbiology and the challenges posed in this research topic. (B) The combination of electrochemistry and ML-based inverse design offers a viable approach to mimicking and controlling the heterogeneities of O₂ and H₂O₂ in microbiology. O, oxidant; R, reductant; E_appl (t), the time-dependent electrochemical voltages applied on electrodes. (C) The design of the electrochemically active microwire array electrodes for the generation of O₂ and H₂O₂ gradients; 4e⁻ ORR & 2e⁻ ORR, four-electron and two-electron oxygen reduction reaction into H₂O and H₂O₂, respectively. (D and E) The 45°-tilting images of SEM for the representative microwire arrays used for the training of the ML model (D) and the ones inversely designed for targeted O₂ and H₂O₂ gradients (E); k = (P, D, L), the morphological vector that includes the P, D, and L of the synthesized wire arrays in units of micrometers. (Scale bars, 20 μm.)Despite recent progress (14–18), there remain major technical challenges, particularly in the achievable spatiotemporal resolution and high-throughput design of concentration profiles to suit a plethora of scenarios in microbiology. Approaches based on microfluidics and hydrogels have been able to achieve concentration gradients of O₂ and H₂O₂ through the provision of either O₂/H₂O₂ source (14, 19–21), O₂/H₂O₂ scavenging agents (15, 22, 23), or a combination of both (24) across liquid-impermeable barriers such as agar layers or polymeric thin films (25, 26). Yet such approaches, dependent on passive mass transport and diffusion across more than 10² μm, are inherently incapable of achieving spatial features of less than 100 μm and temporal resolution smaller than ∼10¹ s, the prerequisites to investigate microbiology at cluster or single-cell levels (10–12). Moreover, the large variations of extracellular O₂ and H₂O₂ gradients in different microbial systems demand an inverse design strategy, which, with minimal expenditure, quickly programs a desired concentration profile catering to a specific biological scenario (2–5). The current lack of inverse design protocol impedes the adoption of controllable extracellular heterogeneity to mimic and investigate microbial systems that are of environmental, biomedical, and sustainability-related significance.We envision that the integration of electrochemically generated concentration gradients with inverse design based on machine learning (ML) will address the aforementioned challenges (Fig. 1B). Electrochemistry offers a venue for transducing electric signals into microscopic concentration profiles within ∼10⁰ μm to ∼10² μm away from electrodes’ surface, following the specific electrode reaction kinetics and the mass transport governing equations in the liquid phase (27). Proper designs of electrodes’ microscopic spatial arrangement and electrochemical kinetics lead to concentration gradients that are spatiotemporally programmable by time-dependent electric signals of varying voltages (28). Such benefits of electrochemically generated concentration gradients lead us to employ electrochemistry as a tool to spatiotemporally control the concentration profiles in the extracellular medium. In one example, we found that wire arrays electrochemically active toward O₂ reduction create anoxic microenvironment about 20 μm away from the aerobic external bulk environments, modulate the size and extent of O₂ depletion in the anoxic microenvironment by the wire array’s morphology and applied electrochemical potential (E_appl), and hence enable O₂-sensitive rhizobial N₂ fixation in ambient air powered by renewable electricity (29). Moreover, while not reported before as far as we know, electrochemically generated concentration heterogeneity is commensurate with ML-based inverse design (30, 31), thanks to the mathematically well-defined electrochemical processes that can be numerically simulated (32, 33). We recently reported neural networks, trained by numerically simulated data, that explore the influence of electrode geometry on electrochemical N₂ fixation and achieve optimized morphologies of wire array electrodes untenable without such an ML-based strategy (34). An inverse design for the electrochemically generated gradients will quickly program desirable microenvironments of O₂ and ROS with high spatiotemporal resolutions, thanks to the well-reported electrochemical transformation related to O₂ and H₂O₂ with high electrochemical selectivity (35, 36).In this work, we report an inverse design based on neural networks for independent electrochemical creation of O₂ and ROS microscopic gradients that are relevant, and mimic their extracellular heterogeneities in microbial systems. We hypothesize that careful design of electrocatalysis of O₂ reduction reaction (ORR) can either facilitate four-electron ORR on Pt electrocatalyst for a controllable O₂ spatiotemporal profile or promote two-electron ORR on Au electrocatalyst for a programmable generation of H₂O₂ gradient without significantly perturbing the O₂ one, thanks to their concentration differences in biological mediums (∼10⁻¹ μM to ∼10¹ μM for H₂O₂ and ∼10¹ μM to ∼10² μM for O₂) (2, 7–11). Electrochemically active microwire array electrodes as exemplary model systems (Fig. 1C) are experimentally shown to achieve tunable heterogeneities of O₂ and H₂O₂ independently, with spatial resolution of ∼10¹ μm and temporal resolution of ∼10° s, and are suitable as a platform for independently perturbing biologically relevant O₂ and H₂O₂ profiles in microbial systems. We further established and experimentally validated two neural networks that inversely design the wire array electrodes’ morphologies toward targeted microenvironments of O₂ and H₂O₂, respectively, which is at least one order of magnitude faster than trial-and-error numerical simulation and two orders of magnitude faster than experimental explorations. The demonstrated inverse design of electrochemically generated controlled gradients not only demonstrates a full electrochemical control of concentration profiles in an electrode’s proximity but also establishes an approach of spatiotemporally mimicking and perturbing extracellular space guided by artificial intelligence.     

Tritiated taurine handling by isolated rat pancreatic islets

A gating of volume-sensitive anion channels may participate in the depolarization of the plasma membrane caused by high concentration of d-glucose in insulin-producing B-cells of the endocrine pancreas. The efflux of tritiated taurine from prelabeled cells is currently used to assess changes in the activity of such channels. The handling of [1,2-³H]taurine by isolated rat pancreatic islets was therefore investigated. The net uptake of [1,2-³H]taurine was found to represent a concentration-, time-, and temperature-dependent process. It was progressively increased in the range of d-glucose concentrations between 2.8 and 8.3 mM, but no further increase was observed at 16.7 mM d-glucose. Over 15 min incubation, the efflux of radioactivity from prelabeled islets was inhibited by MK571 (1.0 mM). It was increased in response to hypoosmolarity both in the presence and absence of extracellular Na⁺. Whether in salt-balanced or Na⁺-deprived media, the efflux of radioactivity from prelabeled islets increased in response to a rise in d-glucose concentration from 2.8 to 5.6 or 8.3 mM, but decreased when the concentration of the hexose was further increased from 8.3 to 16.7 mM. In perifused islets, however, the radioactive efflux from prelabeled islets was inhibited, in a concentration-related manner, when islets first deprived of d-glucose for 45 min were then exposed to 2.8, 5.6, or 16.7 mM d-glucose. Likewise, in prelabeled and perifused islets first exposed for 45 min to 4.0 mM d-glucose, a later rise in hexose concentration to 8.3 mM failed to affect significantly effluent radioactivity, while an increase in hexose concentration from 4.0 to 16.7 mM inhibited the radioactive outflow. In these perifusion experiments, the rise in d-glucose concentration provoked the expected changes in insulin output. The findings obtained in islets examined immediately after preincubation in the presence of [1,2-³H]taurine are consistent with the presence of volume-sensitive anion channels in islet cells and with a gating of such channels in response to a rise in d-glucose concentration from 2.8 to 5.6–8.3 mM. However, the radioactive fractional outflow rate from prelabeled islets seems to reach its highest value at about 8.3 mM d-glucose, being unexpectedly decreased at a higher concentration (16.7 mM) of the hexose. In conclusion, the pleiotropic effects of d-glucose upon tritiated taurine outflow from prelabeled rat islets, which could conceivably be ascribed to differences in the handling of this amino sulfonic acid by distinct islet cell types, indicates that the present approach is far from optimal to characterize unambiguously the regulation by the hexose of volume-sensitive anion channel activity in insulin-producing islet cells.     

Characterization of Al2O3 Matrix Composites Fabricated via the Slip Casting Method Using NiAl-Al2O3 Composite Powder

This work aimed to characterize Al₂O₃ matrix composites fabricated by the slip casting method using NiAl-Al₂O₃ composite powder as the initial powder. The composite powder, consisting of NiAl + 30 wt.% Al₂O₃, was obtained by mechanical alloying of Al₂O₃, Al, and Ni powders. The composite powder was added to the Al₂O₃ powder to prepare the final powder for the slip casting method. The stained composite samples presented high density. EDX and XRD analyses showed that the sintering process of the samples in an air atmosphere caused the formation of the NiAl₂O₄ spinel phase. Finally, the phase composition of the composites changed from the initial phases of Al₂O₃ and NiAl to Al₂O₃, Ni, and NiAl₂O₄. However, in the area of Ni, fine Al₂O₃ particles remaining from the initial composite powder were visible. It can be concluded that after slip casting, after starting with Al₂O₃ and the composite powder (NiAl-Al₂O₃) and upon sintering in air, ceramic matrix composites with Ni and NiAl₂O₄ phases, complex structures, high-quality sintered samples, and favorable mechanical properties were obtained.     

Effect of a hypoglycaemic agent M&B 39890A on glucagon secretion in isolated rat islets of Langerhans

Summary Administration of the compound M&B 39890A lowered serum glucose levels significantly (p<0.001) in genetically obese mice, while no effect on serum insulin levels was observed. In in vitro experiments with isolated rat islets of Langerhans M&B 39890A inhibited arginine-stimulated glucagon release at all concentrations tested (0.5, 5.0 and 50 mol/l). Insulin secretion was not inhibited by M&B 39890A (0.5 and 5.0 mol/l), but was slightly decreased at 50 mol/l. M&B 39890A (5 mol/l) also inhibited glucagon secretion in vitro in the presence of 2 mmol/l, 6 mmol/l and 20 mmol/l glucose, while exerting no effect on insulin secretion. These results suggest that the hypoglycaemic action of M&B 39890A may be due to its direct and selective effect on glucagon secretion; this appears to operate by a mechanism different to that of glucose.     

Pulse Plasma Sintering of NiAl-Al2O3 Composite Powder Produced by Mechanical Alloying with Contribution of Nanometric Al2O3 Powder

NiAl-Al₂O₃ composites, fabricated from the prepared composite powders by mechanical alloying and then consolidated by pulse plasma sintering, were presented. The use of nanometric alumina powder for reinforcement of a synthetized intermetallic matrix was the innovative concept of this work. Moreover, this is the first reported attempt to use the Pulse Plasma Sintering (PPS) method to consolidate composite powder with the contribution of nanometric alumina powder. The composite powders consisting of the intermetallic phase NiAl and Al₂O₃ were prepared by mechanical alloying from powder mixtures containing Ni-50at.%Al with the contribution of 10 wt.% or 20 wt.% nanometric aluminum oxide. A nanocrystalline NiAl matrix was formed, with uniformly distributed Al₂O₃ inclusions as reinforcement. The PPS method successfully consolidated NiAl-Al₂O₃ composite powders with limited grain growth in the NiAl matrix. The appropriate sintering temperature for composite powder was selected based on analysis of the grain growth and hardness of Al₂O₃ subjected to PPS consolidation at various temperatures. As a result of these tests, sintering of the NiAl-Al₂O₃ powders was carried out at temperatures of 1200 °C, 1300 °C, and 1400 °C. The microstructure and properties of the initial powders, composite powders, and consolidated bulk composite materials were characterized by SEM, EDS, XRD, density, and hardness measurements. The hardness of the ultrafine-grained NiAl-Al₂O₃ composites obtained via PPS depends on the Al₂O₃ content in the composite, as well as the sintering temperature applied. The highest values of the hardness of the composites were obtained after sintering at the lowest temperature (1200 °C), reaching 7.2 ± 0.29 GPa and 8.4 ± 0.07 GPa for 10 wt.% Al₂O₃ and 20 wt.% Al₂O₃, respectively, and exceeding the hardness values reported in the literature. From a technological point of view, the possibility to use sintering temperatures as low as 1200 °C is crucial for the production of fully dense, ultrafine-grained composites with high hardness.     

Effect of 2-deoxyglucose on [32P] phosphate and insulin release from perifused rat pancreatic islets

Summary The effect of 2-deoxyglucose on glucose mediated insulin and [³²P]phosphate release was studied by perifusion of isolated rat pancreatic islets. When islets were perifused with media containing 2.8 mmol/l glucose and 20 mmol/l 2-deoxyglucose for 60 minutes and then exposed to media containing 8.3 or 16.7 mmol/l glucose and 20 mmol/l 2-deoxyglucose for the next 15 minutes, insulin release at either glucose concentration was prompt but blunted. Similarly, islets preincubated (90 min) with [³²P] orthophosphate, then perifused with 20 mmol/l 2 deoxyglucose for 75 min and stimulated by either 8.3 or 16.7 mmol/l glucose for the final 15 minutes of 2 deoxyglucose exposure demonstrated obtundation of [³²P]phosphate release. Perifusion of islets with 20 mmol/l 2-deoxyglucose alone induced no heightened³²P efflux. These studies suggest that 2-deoxyglucose affects initial events in stimulus-secretion coupling of glucose mediated insulin release.     

Metabolism of cold-stored pancreatic islets

Summary A previous study showed that the ability of glucose to stimulate insulin release was retained in islets stored at 8 °C for one week provided that glucose was present in a high concentration in the storage medium. The metabolic properties of islets stored in the cold have now been further explored in an attempt to clarify the protective effect of glucose. During storage in the cold the islet formation of ³H₂O from (5-³H) glucose and oxygen consumption were only a few per cent of that of fresh islets whereas the uptake of ⁸⁶Rb⁺ was 20–48%. Rewarming the cold-stored islets to 37 °C after one week of cold-storage restored the ⁸⁶Rb⁺ uptake, the formation of ³H₂O and ¹⁴CO₂ from labelled glucose and oxygen consumption to 75, 80, 60 and 40% respectively of fresh islet levels. The results emphasize the usefulness of cold-storage for preservation of functionally intact isolated islets.On leave from the Metabolic Research Unit, 1143 HSW, University of California, San Francisco, CA 94143, USA     

Alloxan-glucose interaction: Effect on incorporation of14C-leucine into pancreatic islets of rat

Summary The acute effect of alloxan on the incorporation of¹⁴C-leucine into isolated rat islets of Langerhans was studied. I.v. administration of alloxan (40 mg/kg body weight) in rats inhibited the subsequentin vitro incorporation of¹⁴C-leucine into (pro-)insulin in the isolated islets. Glucose (750 mg/kg body weight), when administered 5 min prior to alloxan, completely protected the islets against alloxan toxicity. The protective effect of glucose was partly reversed when the concentration of alloxan was raised to 80 mg/kg body weight. Similar results of inhibition of (pro-)insulin biosynthesis by alloxan and its protection by glucose were obtained when isolated rat islets were exposed to alloxan and/or glucosein vitro. Islets exposed to glucosein vitro immediately after alloxan exposure showed a slower rate of inhibition of (pro-)insulin biosynthesis, as compared to islets washed before exposure to D-glucose. In view of these findings, it is suggested that there is a common recognition site on B-cell for alloxan and glucose. CDRI Communication No. 2617.