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Alloxan cytotoxicity in vitro : Microscope photometric analyses of trypan blue uptake by pancreatic islet cells in suspension

Suspensions of islet cells were prepared by shaking pancreatic islets from non inbred ob/ob mice in a Ca 2+ free buffer. The cells were incubated with or without 20 mM alloxan, and subsequently with Trypan Blue. The uptake of Trypan Blue by cell nuclei was analysed by microscope photometry and by counting the frequency of cells appearing stained on visual inspection. Cells classified as stained o

The pancreatic β cell recognition of insulin secretagogues. XII. Insulin release in response to halogenated hexosamines

The effects of N iodoacetyl 2 amino 2 deoxy (D) glucose and various N bromoacetylglycosylamines on the release of insulin from microdissected pancreatic islets of non inbred ob/ob mice were studied. N Bromoacetyl β (D) glucosylamine (10 m(M)) initiated insulin release in the absence of (D) glucose and, at concentrations of 2.5-10 m(M), but not 20 m(M), potentiated insulin release in response to 10

Preparation and characterization of plasma membrane-enriched fractions from rat pancreatic islets

Methods have been developed for the isolation on a semi-micro scale of a plasma membrane-enriched fraction from rat islets of Langerhans. An important feature of these experiments is the use of 125I-labeled wheat germ agglutinin as a specific probe for plasma membrane-containing fractions. The partly purified plasma membrane fraction had a density in sucrose of about 1.10 and was enriched in the a

The dynamics of insulin release from mouse pancreatic islet cells in suspension

The overall dynamics of glucose-induced insulin release was strikingly similar in dispersed cells and intact islets perifused in parallel. Both preparations exhibited a latency of 1-2 min, after which period there was a brisk rise of insulin release followed by a sustained second phase. During the second phase, insulin release from dispersed cells attained a stable plateau rate, whereas the releas

Studies on the function of pancreatic islet cell membranes.

Pancreatic islets rich in beta-cells were isolated from non-inbred ob/ob-mice and used for studying various aspects of the function of the plasma membrane. A review is given of the authors' work along the following lines: the role of transmembrane transport or membrane binding in the recognition of insulin-releasing sugars, amino acids, sulfonylureas, and sulphydryl-blocking agents; the role of cy

Stimulation of insulin release by thiols

The effects of thiol compounds on insulin release were studied in microdissected pancreatic islets of non-inbred ob/ob mice. In control experiments the reactivity of thiols against 6,6′-dithiodinicotinic acid and the degradation of mouse insulin were measured. At a concentration of 0.1 mM, 1-thio-D-glucose or reduced glutathione potentiated the insulin-releasing action of 10 mM D-glucose without a

Glucagon and insulin release from the allografted canine pancreas

Six previously pancreatectomized dogs were transplanted with ductligated, pancreatic allografts. Glucagon and insulin levels in the venous outflow from the graft and in the systemic circulation were determined during the first 60 minutes after transplantation. Two of the dogs were subjected to L-arginine stimulation 5 days after transplantation and the glucagon levels in the venous outflow from th

The use of dispersed pancreatic islet cells in measurements of transmembrane transport

Suspensions of dispersed islet cells were prepared by shaking collagenaseisolated pancreatic islets of ob ob-mice in Ca2+-free buffer. The dispersed cells exhibited a glucose uptake with stereospecificity for the d isomer and concentrated Rb+ about 30-fold from a medium containing 70 μm RbCl. These results compare well with previous observations on unbroken islets and indicate that the dispersion

The pancreatic β cell recognition of insulin secretagogues : does cyclic AMP mediate the effect of glucose?

Insulin release and the content of cAMP were studied in microdissected pancreatic islets of non inbred ob/ob (obese) mice. In the absence of 3 isobutyl 1 methylxanthine, a phosphodiesterase inhibitor, 20 mM glucose had no effect on cAMP save a very small initial rise detectable by a freeze stop perifusion technique only. However, combined with this methylxanthine, 20 mM glucose produced significan

Cytotoxic effects of a thiol reactive ferritin compound on isolated pancreatic β cells and the protective action of L cysteine

The effects of an electron dense thiol reagent, Hg phenylazo ferritin, on β cell ultrastructure was studied in cell suspensions from mouse pancreatic islets. This type of thiol reagent induced severe damage in the β cells. The ferritin particles were traced with the electron microscope and found to be bound to the plasma membrane as well as to various cytoplasmic structures. The β cell cytotoxic a

Effects of pancreozymin and secretin on insulin release and the role of the exocrine pancreas

Mouse pancreatic islets microdissected free from, or surrounded by, exocrine cells were used to study the effects of secretin and choleeystokinm-pancreozymin on insulin release. Both secretin and cholecystokinin-pancreozymin potentiated glucose-stimulated insulin release regardless of whether exocrine cells were present. The results fail to support the idea that the presence of the exocrine parenc

The preparation of, and studies on, free cell suspensions from mouse pancreatic islets

Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca2+-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200-500 islets, about 2.2×106 cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated f

Assessment of Ultra-Short Heart Variability Indices Derived by Smartphone Accelerometers for Stress Detection

Body acceleration due to heartbeat-induced reaction forces can be measured as mobile phone accelerometer (m-ACC) signals. Our aim was to test the feasibility of using m-ACC to detect changes induced by stress by ultra-short heart rate variability (USV) indices (standard deviation of normal-to-normal interval-SDNN and root mean square of successive differences-RMSSD). Sixteen healthy volunteers wer

Influence of the mutation "diabetes" on insulin release and islet morphology in mice of different genetic backgrounds

Mice, 7-8-mo old, of the C57BL/KsJ-db strain and homozygotic for the mutant gene db, exhibited marked hyperglycemia and moderately elevated serum insulin levels. Light and electron microscopy provided evidence of a slightly decreased proportion of β cells in the pancreatic islets, irregular islet architecture with intraislet ducts, and degenerative as well as hypertrophic changes in the individua

The pancreatic β-cell recognition of insulin secretagogues. VIII. Comparisons of glucose with glyceraldehyde isomers and dihydroxyacetone

d-Glyceraldehyde stimulated the release of insulin from pancreatic islets of Umeå- ob ob-mice whether or not glucose was present in the medium. Like the action of glucose, that of d-glyceraldehyde was biphasic in time, exhibited a sigmoidal dose-response relationship, was potentiated by theophylline, arginine, iodoacetamide, or l-glyceraldehyde, and was inhibited by epinephrine, 2,4-dinitrophenol,

The pancreatic β cell recognition of insulin secretagogues. Influence of neuraminidase treatment on the release of insulin and the islet content of insulin, sialic acid, and cyclic adenosine 3':5' monophosphate

Pancreatic islets rich in β cells and microdissected from noninbred ob/ob mice contained about 15 pmol sialic acid per μg dry weight as determined by a microversion of the Warren thiobarbituric acid method. Removal of half of the sialic acid by treatment with Clostridium perfringens neuraminidase had no effect on the islet content of insulin, moderately inhibited glucose oxidation, and resulted in