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PubMed1981

Subunit composition of mitochondrial F1-ATPase isolated from Saccharomyces carlsbergensis.

1. The subunit stoicheiometry of mitochondrial F1-ATPase from yeast (Saccharomyces carlsbergensis), grown in the presence of [3H]leucine and uniformly labelled [14C]glucose, has been determined. 2. The stoicheiometry on the basis of radioactivity is : alpha: beta: gamma: epsilon = 3 : 3 : 1 : 1. The amount of the smallest subunit, epsilon, could not be measured by this method. 3. The molecular weights of the subunits, determined by urea-SDS gel electrophoresis, are 53 000, 50 000, 33 000, 12 500 and 6500, respectively. The calculated molecular weight of the ATPase is 360 000, assuming the presence of one epsilon subunit per F1. 4. The amino acid composition of the total ATPase and of the individual subunits has been determined. 5. The aurovertin-binding properties of F1 are discussed in relation to the subunit stoicheiometry.

Adenosine Triphosphatases↗

PubMed1986

[Phospholipid biosynthesis in the lungs in a chronic inflammatory bronchopulmonary process].

Glycerokinase, glycerophosphate dehydrogenase, glycerophosphate acyltransferase activity, glycerophosphate, dioxiacetonphosphate level and in vivo incorporation of (U-14C)-glucose into the lung phospholipid structure were studied in normal rats and in conditions of chronic bronchopulmonary inflammation. The inhibition of glycerokinase and glycolytic ways of glycerophosphate formation was demonstrated. The data obtained have shown that glucose incorporation into phosphatidyl cholines, phosphatidyl glycerols and phosphatidyl ethanolamines was decreased, while the incorporation of radioactivity into the sphingomyelins and lysophosphatidyl cholines was significantly activated.

Animals↗

PubMed1977

Failure of lactose and glucose to influence in vivo intestinal calcium transport in normal rats.

The influence of luminal lactose and glucose on intestinal calcium transport was examined in vivo in normal post-weaned young rats. In situ segments of duodenum, midgut and ileum were perfused through the lumen for 1 h with an isotonic solution of 3.4 mM calcium containing radioactive isotopic 45Ca as tracer and additionally either 10 mM lactose or 10 mM glucose, or, for the control group, no saccharide. Net absorption was measured by disappearance of luminal calcium; lumen-to-plasma flux was determined from the disappearance of tracer 45Ca. This concentration of luminal lactose or glucose had no direct effect on calcium transport under these steady state conditions.

Animals↗

PubMedDate not supplied

Dissociation of insulin absorption and blood flow during massage of a subcutaneous injection site.

Nine healthy volunteers with normal body weights were injected subcutaneously with 125I-labeled soluble human insulin (10 U) in one thigh and 133Xe in the contralateral thigh for the measurement of subcutaneous blood flow on 2 consecutive mornings. On one of the days, standardized massage of both injection sites was performed for 30 min starting 30 min after insulin injection. Serum insulin and plasma glucose were determined intermittently before, during, and after massage, and elimination of radioactivities was monitored continuously by external detectors. During massage, the first-order elimination rate constants of 125I increased approximately sixfold compared with the rise during control (0.19 +/- 0.04 to 0.88 +/- 0.15%/min during the last 15 min of the massage vs. 0.21 +/- 0.03 to 0.32 +/- 0.03%/min during control). Serum insulin increased from 13.8 +/- 1.8 mU/L before massage to a maximal value of 56.4 +/- 8.7 mU/L 10 min after massage (vs. 15.3 +/- 3.0 and 19.7 +/- 2.2 mU/L during control). Plasma glucose fell significantly faster on the massage day, from 10 min after massage onward. No significant alteration in the subcutaneous blood flow was found during or after massage. The results suggest that the pronounced enhancement of insulin absorption induced by massage of the injection site is mainly not blood flow mediated.

Absorption↗

PubMed1986

Studies of anaerobic and aerobic glycolysis in Saccharomyces cerevisiae.

Glucose metabolism was followed in suspensions of Saccharomyces cerevisiae by using 13C NMR and 14C radioactive labeling techniques and by Warburg manometer experiments. These experiments were performed for cells grown with various carbon sources in the growth medium, so as to evaluate the effect of catabolite repression. The rate of glucose utilization was most conveniently determined by the 13C NMR experiments, which measured the concentration of [1-13C]glucose, whereas the distribution of end products was determined from the 13C and the 14C experiments. By combining these measurements the flows into the various pathways that contribute to glucose catabolism were estimated, and the effect of oxygen upon glucose catabolism was evaluated. From these measurements, the Pasteur quotient (PQ) for glucose catabolism was calculated to be 2.95 for acetate-grown cells and 1.89 for cells grown on glucose into saturation. The Warburg experiments provided an independent estimate of glucose catabolism. The PQ estimated from Warburg experiments was 2.9 for acetate-grown cells in excellent agreement with the labeled carbon experiments and 4.6 for cells grown into saturation, which did not agree. Possible explanations of these differences are discussed. From these data an estimate is obtained of the net flow through the Embden-Meyerhof-Parnas pathway. The backward flow through fructose-1,6-bisphosphatase (Fru-1,6-P2-ase) was calculated from the "scrambling" of the 13C label of [1-13C]glucose into the C1 and C6 positions of trehalose. Combining these data allowed us to calculate the net flux through phosphofructokinase (PFK). For acetate-grown cells we found that the relative flow through PFK is a factor of 1.7 faster anaerobically than aerobically.(ABSTRACT TRUNCATED AT 250 WORDS)

Aerobiosis↗

PubMed1976

Effects of glucose on 45Ca2+ uptake by pancreatic islets as studied with the lanthanum method.

1. Fluxes of 45Ca2+ were studied in pancreatic islets from non-inbred ob/ob-mice. Because La3+ blocked the transmembrane fluxes of 45Ca2+ in islet cells, incubations aimed at measuring glucose-induced changes of the intracellular Ca2+ were ended by washing the islets with 2 mM-La3+ for 60 min. 2. Uptake of 45Ca2+ progressed for 2 hr; the intracellular concentration of exchangable Ca2+ was about 7 m-mole/kg dry wt., as estimated from the isotope distribution at apparent equilibrium in islets exposed to 3 mM D-glucose. Raising the D-glucose concentration to 20 mM enhanced the 45 Ca2+ uptake whether or not the islets had first been equilibrated with the isotope. The stimulatory effect of D-glucose was observed in Tris buffer containing no anions but Cl- as well as in polyanionic bicarbonate buffer. The effect could not be reproduced with equimolar L-glucose. 3. The rate of 45Ca2+ release was the same whether the islets had been pre-loaded in the presence of 3 or 20 mM D-glucose. Thus the 45Ca2+ that had been taken up in response to 20 mM D-glucose appeared to be released much more slowly than the bulk of intracellular 45Ca2+. The release of 45Ca2+ was not significantly influenced by D-glucose during the release period. Incubation for 30 min was require for half of the radioactivity to be released. 4. The rates of insulin secretion were about the same in uni-anionic Tris buffer as in polyanionic bicarbonate buffer. A marked insulin secretory response to 20 mM D-glucose was observed in either buffer. 5. It is concluded that 20 mM D-glucose causes a net uptake of Ca2+ from the extracellular fluid into the interior of the beta-cells. This uptake is probably not regulated at the level of the plasma membrane but more likely reflects an increased affinity of some intracellular phase or compartment for the ion. Because the observed uptake and release of intracellular 45Ca2+ are slow processes in comparison with the rapid effects of extracellular Ca2+ on insulin secretion, insulin secretion may also depend on a more superficial and La3+-displacable Ca2+ pool.

Animals↗

PubMed2001

An extract of Lagerstroemia speciosa L. has insulin-like glucose uptake-stimulatory and adipocyte differentiation-inhibitory activities in 3T3-L1 cells.

The effects of extracts isolated from Lagerstroemia speciosa L. (banaba) on glucose transport and adipocyte differentiation in 3T3-L1 cells were studied. Glucose uptake-inducing activity of banaba extract (BE) was investigated in differentiated adipocytes using a radioactive assay, and the ability of BE to induce differentiation in preadipocytes was examined by Northern and Western blot analyses. The hot water BE and the banaba methanol eluent (BME) stimulated glucose uptake in 3T3-L1 adipocytes with an induction time and a dose-dependent response similar to those of insulin. Furthermore, there were no additive or synergistic effects found between BE and insulin on glucose uptake, and the glucose uptake activity of insulin could be reduced to basal levels by adding increasing amounts of BE. Unlike insulin, BE did not induce adipocyte differentiation in the presence of 3-isobutyl-1-methylxanthine (IBMX) and dexamethasone (DEX). BE inhibited the adipocyte differentiation induced by insulin plus IBMX and DEX (IS-IBMX-DEX) of 3T3-L1 preadipocytes in a dose-dependent manner. The differences in the glucose uptake and differentiation inhibitory activities between untreated cells and those treated with BE were significant (P < 0.01). The inhibitory activity was further demonstrated by drastic reductions of peroxisome proliferator-activated receptor gamma2 (PPARgamma2) mRNA and glucose transporter-4 (GLUT4) protein in cells induced from preadipocytes with IS-IBMX-DEX in the presence of BE. The unique combination of a glucose uptake stimulatory activity, the absence of adipocyte differentiation activity and effective inhibition of adipocyte differentiation induced by IS-IBMX-DEX in 3T3-L1 cells suggest that BE may be useful for prevention and treatment of hyperglycemia and obesity in type II diabetics.

1-Methyl-3-isobutylxanthine↗

PubMed1987

Effects of glucose transport on the incorporation of 32P-inorganic phosphate into phospholipids in Hymenolepis diminuta (Cestoda).

1. The absorption of glucose and alpha-methyl-D-glucoside by Hymenolepis diminuta was concentration dependent. 2. alpha-Methyl-D-glucoside competitively inhibited glucose absorption and was not metabolised by the parasite. 3. alpha-Methyl-D-glucoside significantly lowered (P less than 0.05) the incorporation of 32Pi into all phospholipid classes. 4. Glucose inhibited (P less than 0.01) 32Pi incorporation into phosphatidylcholine only. 5. Phlorizin did not affect 32Pi labelling of phospholipids. 6. Serotonin and histamine stimulated (P less than 0.01) 32Pi labelling of all phospholipid classes. 7. Radioactivities in water soluble fractions were increased (P less than 0.01) in the presence of glucose, serotonin and histamine.

Animals↗

PubMed1997

Kinetics of Sugar Transport and Phosphorylation Influence Glucose and Fructose Cometabolism by Zymomonas mobilis.

The competitive inhibition of fructokinase by glucose has been proposed as the mechanism by which Zymomonas mobilis preferentially consumes glucose from mixtures of glucose and fructose and accumulates fructose when growing on sucrose. In this study, incorporation of radioactive fructose into biomass was used as a measure of fructose catabolism. It was determined that the rate of fructose incorporation by Z. mobilis CP4 was somewhat lower in the presence of an equimolar concentration of glucose but that the inhibition of fructokinase by glucose was not nearly as severe in vivo as was predicted from in vitro studies. Interestingly, addition of glucose to a culture of Z. mobilis CP4-M2, a glucokinaseless mutant, resulted in an immediate and nearly complete inhibition of fructose incorporation. Furthermore, addition of nonmetabolizeable glucose analogs had a similar effect on fructose catabolism by the wild-type Z. mobilis CP4, and fructose uptake by Z. mobilis CP4-M2 was shown to be severely inhibited by equimolar amounts of glucose. These results suggest that competition for fructose transport plays an important role in preferential catabolism of glucose from sugar mixtures. Indeed, the apparent K(infm) values for sugar uptake by Z. mobilis CP4 were approximately 200 mM for fructose and 13 mM for glucose. Other experiments supported the conclusion that a single facilitated diffusion transport system, encoded by the glf gene, is solely responsible for the uptake of both glucose and fructose. The results are discussed with regard to the hypothesis that the kinetics of sugar transport and phosphorylation allow the preferential consumption of glucose and accumulation of fructose, making the fructose available for the enzyme glucose-fructose oxidoreductase, which forms sorbitol, an important osmoprotectant for Z. mobilis when growing in the presence of high sugar concentrations.

Journal Article↗

PubMed1978

Collagenase isolation and 45Ca efflux studies of human islets of Langerhans.

22 human pancreases were removed soon after circulatory arrest from donors aged 15--63 years. Part of the pancreas was used for isolation of islets by the collagenase technique. The mean yield +/- SEM, expressed as the number of islets isolated from 4 g pancreas was 79 +/- 15. The yield varied considerably even when the pancreas was removed immediately after circulatory arrest and the histology was normal, and there was no correlation with the age of the donors. The islet content of insulin (ng/microgram dry islet) was 8.5 +/- 1.2 (mean +/- SEM). Isolated islets were loaded with 45Ca in the presence of 20 mM glucose and placed in a perfusion apparatus for further studies of the 45Ca washout. The decrease of washout of radioactivity in Ca2+-deficient medium offers support for the existence of a Ca2+-Ca2+ exchange process. When added in a concentration of 20 mM, glucose tended to stimulate 45Ca efflux in perfusion medium of ordinary ionic composition but inhibited this process when the medium was deficient in Ca2+. Exposure to the calcium ionophore X-537A resulted in immediate stimulation of of 45Ca efflux from human islets as previously observed for islets from rats and mice. This suggests that Ca2+ has a direct regulatory role for insulin release also in humans.

Adolescent↗

PubMedDate not supplied

Protein secretion by the rat epididymis can be selectively modified in vitro by local anesthetics, glucose deprivation, dinitrophenol, ouabain, and ionophores.

Protein secretion by the caput epididymidis has been examined in vitro using radioactive methionine as a precursor for protein synthesis. Newly synthesized and secreted proteins were separated by gel electrophoresis and visualized by fluorography. Local anesthetics such as procaine had the ability to reduce the secretion of some, but not all, proteins. Selective inhibition of secretion of the same proteins occurred when either dihydrocytochalasin B, monensin, ouabain, or dinitrophenol was added to the medium, or when the concentration of glucose was reduced below 1 mM. Calcium ionophore also selectively modified protein secretion, but the proteins affected were different from those influenced by local anesthetics. Other agents tested (eg, adrenergic and cholinergic agonists and antagonists, sodium pentobarbitone, antipsychotic drugs, cyclic AMP, colchicine) did not selectively modify protein secretion, even though overall protein synthesis and secretion was reduced in some instances. Procaine and dihydrocytochalasin B also reduced glucose utilization by epididymal tissue and it is suggested that these agents may reduce protein secretion by limiting the supply of energy for the exocytotic process. This conclusion is supported by the fact that dinitrophenol, an uncoupler of oxidative phosphorylation, caused a similar alteration in protein secretion. The possibility that a restricted energy supply modifies protein secretion primarily by creating a disturbed intracellular Na/K balance is suggested by the observation that the monovalent ionophore, monensin, and the Na/K ATPase inhibitor, ouabain, were both able to duplicate the effects of the local anesthetics.

2,4-Dinitrophenol↗

PubMed1966

Chemical degradation of dipicolinic acid-C14 and its application in biosynthesis by Penicillium citreo-viride.

Kanie, Matuso (Kagoshima University, Kagoshima, Japan), Shigeo Fujimoto, and J. W. Foster. Chemical degradation of dipicolinic acid-C(14) and its application to biosynthesis by Penicillium citreo-viride. J. Bacteriol. 91:570-577. 1966.-A chemical degradation of dipicolinic acid-C(14) has been worked out, enabling determination of the specific radioactivity of the carboxyl-carbons (carbons-7 and -8), and of the following carbons of the pyridine ring: carbons-2 and -6 combined, carbons-3 and -5 combined, and carbon-4. The degradation was applied to dipicolinic acid synthesized by washed, submerged mycelium of the mold from glucose and C(14)O(2), and from glucose-1-C(14), -2-C(14), and -6-C(14). The distribution of radioactivity within the labeled dipicolinic acids is consistent with operation of respiratory cycles and with the incorporation of one molecule of CO(2) in the pyridine acid. A C(3) compound is a primary building block. The C(7) chain is believed to result from a C(3) plus C(4) condensation, pyruvic acid and aspartic acid beta-semialdehyde being proposed as likely precursors. Other aspects of the biosynthesis of C(7) open-chain compounds and of dipicolinic acid are discussed.

Carbon Dioxide↗

PubMed1981

Glucose utilization during chromatolysis: a 14C deoxyglucose study.

After transection of the facial and hypoglossal nerves, rats were injected with 14C 2-deoxyglucose. Contact-autoradiographs of histological sections showed increased radioactivity in the motor nuclei of the operated side between 1 and 28 days post operation. This suggests an increase of glucose utilization and thus an enhanced energy supply to the facial and hypoglossal nuclei during the process of axonal reaction.

Animals↗

PubMed2001

Cerebral glucose transport implies individualized glial cell function.

Previous positron emission tomography (PET) measurements of cerebral glucose transport using [11C]-3-O-methylglucose (CMG) suggested an interindividual variation in the values of the rate constant of tracer outflow (k2) larger than that for the clearance rate of inflow (K1). These two parameters were examined in healthy cerebral cortex by dynamic PET in 4 men and 2 women (aged 24 to 73 years) without neurologic disease, and in 1 man (42 years) with a recent left hemispheric cerebral infarction under normoglycemia (average blood plasma d-glucose concentration, 5.44 +/- 1.94 micromol/mL) and again under hyperglycemia (average, 10.24 +/- 1.44 micromol/mL). Time-radioactivity curves were obtained from healthy cortex (grey matter) and plasma and analyzed for the values of K1 and k2 by two graphical approaches and two fitting procedures. Both K1 and k2 significantly declined with increasing plasma glucose levels. A highly significant interindividual but not intraindividual variability for k2 was found at normoglycemia and hyperglycemia. The interindividual variability of K1, although borderline significant, was less than that of k2. Accordingly variable were the distribution volumes K1/k2. These data suggest individualized glial cell function and may be relevant to pathogenesis of neuropsychiatric disease.

3-O-Methylglucose↗

PubMed1991

De novo synthesis of diacylglycerol from glucose. A new pathway of signal transduction in human neutrophils stimulated during phagocytosis of beta-glucan particles.

The phagocytosis of beta-glucan particles by human neutrophils and the associated activation of NADPH O2- forming oxidase were accompanied by an increased hydrolysis of phosphoinositides by phospholipase C, hydrolysis of phosphatidylcholine by phospholipase D, accumulation of diglyceride (DG) mass, and [Ca2+]i rise. The reaction of phospholipid hydrolysis played a minor role in the formation of DG, which was mainly formed by de novo synthesis from glucose. The activation of this pathway was shown by the stimulation of the incorporation of [U-14C]glucose into DG, which occurred very rapidly after the challenge of neutrophils with beta-glucan particles. This DG derived from glucose was found almost completely as 1-acyl-2-acyl-glycerol (DAG). On the basis of the finding that phosphatidic acid was the precursor of DAG, an increase in the incorporation of [U-14C]acetate into DAG did not occur, and the [14C]radioactivity was in the glycerol backbone, the synthesis of DAG from [U-14C]glucose occurred very likely via dihydroxyacetone phosphate and glycerol 3-phosphate, stepwise acylation to phosphatidic acid, and dephosphorylation by phosphatidate phosphatase.

Calcium↗

PubMed1987

Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase activity.

Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase [(Na,K)-ATPase] activity (HK cells) was compared with that in normal canine erythrocytes (LK cells). Activities of some of the glycolytic enzymes in the HK cells were significantly higher than those in LK cells. The concentrations of adenosine triphosphate (ATP) and glycolytic intermediates in HK cells were almost equal to those in LK cells. Glucose utilization and lactate production by HK cells in vitro and incorporation of [32P]orthophosphate or [14C]glucose into 2,3-diphosphoglycerate in HK cells were higher than in LK cells. Radioactivity of [32P]ATP in HK cells was lower than in LK cells, but increased to approximately that of LK cells when (Na,K)-ATPase of HK cells was completely blocked by ouabain. When HK cells and LK cells were incubated in the absence of glucose, the concentration of ATP in HK cells was decreased more than that of LK cells. Although ouabain reduced the rate of decrease in ATP in HK cells, the decrease in ATP in HK cells was still 2-fold that in LK cells. The half-life of HK cells was about one-half that of LK cells. The results indicated that glycolysis is greater in HK cells than in LK cells, and that the increased glycolysis in HK cells was stimulated by an increased rate of ATP breakdown for active cation transport by the (Na,K)-ATPase and by increased degradation of ATP for some other pathway, eg, glutathione synthesis. Thus, the increased demand for ATP in HK cells might result in shortening the lifespan of HK erythrocytes.

Animals↗

PubMed1990

Mechanism and pathway of glucose transportation in a digenetic trematode Ceylonocotyle scoliocoelium.

The mode of absorption, kinetics and mechanism of transportation of 14C--glucose in a digenetic trematode, Ceylonocotyle scoliocoelium were studied by culturing the trematodes under in vitro condition in Hank's saline supplemented with radioactive tracer. It was then followed separately by autoradiography and liquid scintillation. Autoradiographic study reveals that the maximum amount of glucose uptake occurs through cuticle whereas the role of intestinal canal is insignificant in this respect. Maximum cpm is recorded in the precipitate followed by TCA treatment. This is due to the presence of a protein-bound 14C-glucose which is assumed as the carrier protein. The inhibitor, phlorizin, used here presumably acts noncompetitively.

Absorption↗

PubMed1998

Fetal vascular responses to maternal glucose administration in streptozocin-induced ovine diabetes mellitus.

OBJECTIVE: To assess the impact of administration of a glucose load to the pregnant diabetic ewe on fetal biochemistry and umbilical-placental perfusion. METHODS: Six pregnant ewes were rendered glucose intolerant by the administration of streptozocin at 85-90 days gestation. An intravenous glucose load was administered to the 6 diabetic ewes and 4 control animals at 125-130 days gestation. Assessment of the fetal responses to this glucose load was made by cardiovascular monitoring, biochemical evaluation and measurement of fetal blood flows by the radioactive microsphere technique. RESULTS: In the fetus of the diabetic ewe the arterial pH progressively decreased from 7.36 +/- 0.04 to 7.10 +/- 0.09 (p < 0.05) 180 minutes after the maternal glucose load. These fetuses displayed a consistent and significant reduction in arterial oxygen tension compared to controls, with a decrease in pO2 from 14.9 +/- 3.3 mmHg at baseline to 11.3 + 3.7 mmHg after 180 minutes. Umbilical-placental blood flow did not alter significantly in the fetus of the diabetic ewe following glucose administration, although in the control fetuses there was a significant reduction from baseline after 60 minutes. Blood flow to the fetal brain and kidney were significantly elevated in the fetus of the diabetic ewe compared to control at all times, suggesting a metabolic adaptation to preserve oxygen delivery may be present. CONCLUSIONS: Maternal hyperglycemia produces profound metabolic alterations in the ovine fetus which are unaccompanied by significant alterations in umbilical-placental blood flow. In the chronically hyperglycemic pregnant ewe, a glucose load produces a major disturbance of fetal homeostasis.

Animals↗