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PubMed1972

Effect of Sendai virus infection on lipid metabolism in chick embryo fibroblasts.

Lipid metabolism in the "abortive" system, Sendai virus-infected primary chick embryo fibroblasts, was examined by using (32)P-orthophosphate, (14)C-glucose, and (14)C-glycerol as precursors. Incorporation of radioactivity from (32)P-orthophosphate and (14)C-glucose into lipid was increased in infected cells. Synthesis of all individual phospholipids was about equally stimulated. There was also evidence for increased lipid synthesis in more productively infected monkey kidney cells. Incorporation of (14)C-glycerol when at a high level in the medium was also increased. However, when this precursor was supplied in minute quantities of high specific activity, incorporation was inhibited. Even though incorporation of radioactivity from (14)C-glucose was stimulated during long labeling periods, the uptake of this precursor during short pulses was inhibited in infected cells. The phenomenon of increased labeling of triglyceride in infected chick cells under certain conditions is discussed, in conjunction with the other effects, in terms of related changes in other virus-infected systems.

Animals↗

PubMed1975

Regulation of carbohydrate metabolism in lymphoid tissue. Quantitative aspects of [U-14C]glucose oxidation by rat spleen slices.

When washed spleen slices from fed rats are incubated with 3 mm-[U-14C]glucose, the rate of glucose utilization (46.2 mumol/h per g dry wt.) is sufficient to account, theoretically, for 80% of the O2 consumption. Measurement of net lactate production, however, and the fate of the radioactive carbon, indicates that the contribution of glucose to the respiratory fuel of the tissue is only 25-30% whereas 60-70% of the glucose utilized is converted into lactate. At saturating glucose concentrations (above 5 mm) its contribution to the respiratory fuel of the slice is increased to a maximum value of 34-39%. Only 2% of the glucose utilized is metabolized via the oxidative steps of the pentose phosphate pathway. Starvation for 72 h marginally increases both the rate of glucose utilization (by 21%) and its net contribution to the respiratory fuel (by 29%). Insulin, glucagon, adrenaline and adenosine 3':5'-cyclic monophosphate have no significant effect on either the rate of glucose utilization or on the pattern of radioactive isotope distribution. The uptake of glucose is increased by only 20%, whereas the production of lactate doubles when slices are incubated under anaerobic conditions. In assessing the suitability of spleen slices for metabolic studies, the only serious major perturbation, compared with the freeze-clamped organ, is an elevated mitochondrial [NAD+]/[NADH] ratio (connected with increased endogenous NH3 production) that is partially restored to normal values on incubation with glucose. Equal proportions of erythrocytes and leucocytes are found in the washed spleen slice. Metabolic contributions of the constituent cell populations in the washed slice are calculated and it is concluded that lymphocytes account for the major part of the glycolytic metabolism (80-90%), whereas the contribution of erythrocytes is insignificant.

Adenosine Diphosphate↗

PubMed1980

Intestinal absorption of 14C-labelled beta-cyclodextrin in rats.

beta-Cyclodextrin and glucose labelled universally by 14C was administered orally to rats and blood radioactivity level was followed. In the case of glucose about 20% of the administered radioactivity, related to 10 ml blood was to be found and this appeared in the blood within 10 min following the oral administration. In the case of beta-cyclodextrin maximum 5% of the administered activity could be found in blood, and even this only 4 to 10 h following administration. Presumably, beta-cyclodextrin cannot be absorbed either from the stomach or from the small intestine, only the labelled open-chain dextrins and glucose formed from cyclodextrins on amylase action. Therefore it is very likely that the orally administered beta-cyclodextrin cannot provoke toxic symptoms.

Animals↗

PubMed1989

Glucose metabolism and NADH recycling by Treponema hyodysenteriae, the agent of swine dysentery.

Glucose metabolism and the mechanisms of NADH oxidation by Treponema hyodysenteriae were studied. Under an N2 atmosphere, washed cell suspensions of the spirochete consumed glucose and produced acetate, butyrate, H2, and CO2. Approximately twice as much H2 as CO2 was produced. Determinations of radioactivity in products of [14C]glucose and [14C]pyruvate metabolism and analyses of enzyme activities in cell lysates revealed that glucose was catabolized to pyruvate via the Embden-Meyerhof-Parnas pathway. The results of pyruvate exchange reactions with NaH14CO3 and Na14COOH demonstrated that pyruvate was converted to acetyl coenzyme A (acetyl-CoA), H2, and CO2 by a clostridium-type phosphoroclastic mechanism. NADH:ferredoxin oxidoreductase and hydrogenase activities were present in cell lysates and produced H2 from NADH oxidation. Phosphotransacetylase and acetate kinase catalyzed the formation of acetate from acetyl-CoA. Butyrate was formed from acetyl-CoA via a pathway that involved 3-hydroxybutyryl-coenzyme A (CoA) dehydrogenase, butyryl-CoA dehydrogenase, and butyryl-CoA transferase. T. hyodysenteriae cell suspensions generated less H2 and butyrate under 10% O2-90% N2 than under 100% N2. Cell lysates contained NADH oxidase, NADH peroxidase, and superoxide dismutase activities. These findings indicated there are three major mechanisms that T. hyodysenteriae cells use to recycle NADH generated from the Embden-Meyerhof-Parnas pathway--enzymes in the pathway from acetyl-CoA to butyrate, NADH:ferredoxin oxidoreductase, and NADH oxidase. Versatility in methods of NADH oxidation and an ability to metabolize oxygen could benefit T. hyodysenteriae cells in the colonization of tissues of the swine large bowel.

Animals↗

PubMed2002

Effects of D-mannoheptulose upon D-glucose metabolism in pancreatic B and non-B islet cells.

D-mannoheptulose was recently proposed to be transported into cells mainly at the intervention of GLUT2. In the present study, the heptose (10 mM) decreased the steady state content of dispersed rat pancreatic islet cells in D-[U-(14)C]glucose, and inhibited to a greater relative extent the utilization of D-[5-(3)H]glucose, the oxidation of D-[U-(14)C]-glucose and its conversion to radioactive amino acid when the dispersed islet cells were incubated at 16.7 mM rather than 2.8 mM D-glucose. A comparable situation was found in purified islet B-cells, whereas D-mannoheptulose only exerted minor to negligible effects upon the metabolism of D-glucose in non-B islet cells. This coincided with a much higher uptake of D-[(3)H]mannoheptulose by B, as distinct from non-B, islet cells. These findings indicate that the unexpectedly greater relative inhibitory action of D-mannoheptulose upon D-glucose metabolism by isolated islets (or dispersed islet cells) observed at high rather than low hexose concentration cannot be accounted for solely by differences in the relative contribution of non-B cells to total D-glucose metabolism by islets incubated at increasing concentrations of D-glucose. A comparable metabolic response to D-mannoheptulose is indeed observed in purified B cells. It could be attributable, in part at least, to D-glucose and D-mannoheptulose countertransport, resulting inter alia in a greater net uptake of the heptose by B cells exposed to a high concentration of the hexose.

Animals↗

PubMed1988

Analysis of the reaction products from incubation of sugarcane vacuoles with uridine-diphosphate-glucose: no evidence for the group translocator.

Isolated sugarcane (Saccharum spp. hybrid H50-7209) vacuoles incorporate radioactivity during incubation with labeled UDP-glucose by a mechanism which was postulated to be responsible for sucrose storage in the vacuoles (UDP-glucose group translocator). Analysis of the reaction products in the medium revealed that several enzymic processes are going on during incubation with UDP-glucose such as production of hexose phosphates, UMP, and sugars, all of which seem unrelated to the incorporation of radioactivity into vacuoles. The incorporated radioactivity was identified mainly as (1-->3)-beta-glucan (callose) of polymerization grades up to more than 20. Callose occurs as a contaminant at the surface of isolated vacuoles coming from the plasmalemma. The properties of UDP-glucose incorporation into the vacuolar preparation compared favorably with known properties of callose synthase. The low mol wt glucans that are found are probably degradation products of labeled callose due to hydrolases, which are liberated by centrifugation of vacuoles. The labeled disaccharide, which chromatographically had been formerly identified as sucrose, is laminaribiose. No sucrose (or sucrose phosphate) could be identified in the vacuole preparation after incubation with UDP-glucose. Thus, the mechanism of sucrose storage in sugarcane vacuoles is still open.

Journal Article↗

PubMed1978

[Simultaneous enzyme determination of C-14 glucose and C-14 fructose in human body fluids].

A new enzymatic method for simultaneous estimation of radioactivity of 1-14C-glucose and 1-14C-fructose is described. It is based on the isomerisation of 1-14C-fructose to 1-14C-glucose by phosphoglucoseisomerase (EC 5.3.1.9) and on its enzymatic removal as ribulose-5-phosphate and 14CO2. The method is specific, reproducible, and gives over 97% recoveries of glucose and fructose concentrations up to 11 mmol/1 in aquos and biological solutions.

Body Fluids↗

PubMed2006

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 concentrations 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.

Animals↗

PubMed2002

Non-invasive measurements of myocardial carbon metabolism using in vivo 13C NMR spectroscopy.

Despite their prime role in maintaining contractile performance, myocardial substrate uptake, substrate preference and metabolism are difficult to assess non-invasively. The objective of the present work was to extend the scope of cardiac 13C nuclear magnetic resonance (NMR) spectroscopy to the in vivo situation ('closed-chest model') and to quantitatively appraise myocardial metabolism in vivo. For this purpose, overnight-fasted Sprague-Dawley rats received intravenous infusions of non-radioactive 13C-labeled glucose, 3-hydroxybutyrate, and acetate as markers for glycolysis, metabolism of ketone bodies and direct incorporation into tricarboxylic acid (TCA) cycle, respectively. In vivo 13C NMR spectra (at 7 T) were acquired from the myocardium with a time resolution of 6 min. At the end of the infusion experiments, tissue extracts were prepared and further analyzed by high-resolution 13C NMR spectroscopy in order to corroborate the findings obtained in vivo. Accordingly, 3-hydroxybutyrate and acetate were rapidly extracted by the myocardium and supplied 42 +/- 6 and 53 +/- 9% of the acetyl-CoA for TCA cycle operation, whereas glucose, although also well extracted, did not contribute to myocardial oxidative metabolism. Myocardial TCA cycle turnover (V(TCA)) in vivo was estimated at 1.34 +/- 0.07 micromol/min/g wet weight, myocardial oxygen consumption (MVO2) at 2.95 +/- 0.16 micromol/min/g wet weight, exchange rate between alpha-ketoglutarate and glutamate (V(x)) at 1.22 +/- 0.08 micromol/min/g wet weight and rate of glutamine synthesis (V(gln)) at 0.14 +/- 0.02 micromol/min/g wet weight. The substantial synthesis of myocardial glutamine is in contrast to experiments with isolated and saline perfused hearts. In conclusion, it is demonstrated that 13C NMR spectroscopy of the heart in intact rats is feasible and provides new quantitative insight into myocardial substrate uptake, preference and metabolism in vivo.

3-Hydroxybutyric Acid↗

PubMed1977

Evidence for a specific transport of D-hexoses across the human term placenta in vitro.

Isolated cotylidons of human term placentas are perfused artificially on the fetal and maternal side. The relative transfer rates of radioactive labelled D-glucose, L-glucose, D-mannose and D-mannitol across the placenta are measured and the inhibition of these transports by phloretin is studied: 1. Transfer rates of D-glucose and D-mannose exceed that of L-glucose about 1.5-4 times. 2. Phloretin (10(-3) mol/l) decreases the transports of D-hexoses, whereas the transport of L-GLUCOSE REMAINS UNAFFECTED. 3. If the concentration of D-glucose is increased, the transport of D-mannose is inhibited competitively. 4. L-glucose and D-mannitol equal in transfer rates. These results show, that D-hexoses cross the human placenta by a specific carrier and by simple diffusion.

Biological Transport, Active↗

PubMed1990

Dichloroacetate increases glucose use and decreases lactate in developing rat brain.

Dichloroacetate (DCA) activates pyruvate dehydrogenase (PDH) by inhibiting PDH kinase. Neutralized DCA (100 mg/kg) or saline was intravenously administered to 20 to 25-day-old rats (50-75g). Fifteen minutes later a mixture of [6-14C]glucose and [3H]fluorodeoxyglucose (FDG) was administered intravenously and the animals were sacrificed by microwave irradiation (2450 MHz, 8.0 kW, 0.6-0.8 sec) after 2 or 5 min. Brain regional rates of glucose use and metabolite levels were determined. DCA-treated rats had increased rates of glucose use in all regions studied (cortex, thalamus, striatum, and brain stem), with an average increase of 41%. Lactate levels were lower in all regions, by an average of 35%. There were no significant changes in levels of ATP, creatine phosphate, or glycogen in any brain region. Blood levels of lactate did not differ significantly between the DCA- and the saline-treated groups. Blood glucose levels were higher in the DCA group. In rats sacrificed by freeze-blowing, DCA treatment caused lower brain levels of both lactate and pyruvate. These results cannot be explained by any systemic effect of DCA. Rather, it appears that in the immature rat, DCA treatment results in activation of brain PDH, increased metabolism of brain pyruvate and lactate, and a resulting increase in brain glycolytic rate.

Animals↗

PubMed1981

Regulation of carbohydrate metabolism in cultured mammalian cells: energy provision in a glycolytic mutant.

The metabolism of radioactively labelled D-glucose, L-glutamine, and L-glutamate has been determined in a glycolytic mutant of Chinese-hamster ovary cells, R1.1.7, and in its parent, CHO-K1. The complete oxidation of glucose via the TCA-cycle is negligible in both cell types, but there is significant oxidation of carbon-1. CHO-K1 cells derive most of their energy from glycolysis and are independent of respiration in the short term. R1.1.7 cells are respiration-dependent and are rapidly killed by respiratory inhibitors. Both cell types oxidize L-glutamine and L-glutamate, but oxidation of these substrates does not appear sufficient to satisfy completely the energy requirements of R1.1.7 cells.

Anemia, Hemolytic, Congenital Nonspherocytic↗

PubMed1992

Glucose tolerance and insulin secretion in spontaneously hyperthyroid cats.

Glucose tolerance and insulin secretion after administration of a glucose load were determined in 11 clinically normal cats and 15 cats with spontaneous hyperthyroidism. In six hyperthyroid cats, a glucose tolerance test was repeated after treatment with radioactive iodine (131I). All cats had similar baseline glucose concentrations. However, the cats with hyperthyroidism had a significantly decreased glucose clearance, which was worse after treatment. Hyperthyroidism also caused a marked increase in basal and glucose-stimulated insulin secretion, which was not improved with treatment. It is concluded that hyperthyroidism in cats may lead to long-lasting alterations of glucose tolerance and insulin secretion which may not be reversed by treatment.

Animals↗

PubMed2005

Sucrose utilization during somatic embryo development in black spruce: involvement of apoplastic invertase in the tissue and of extracellular invertase in the medium.

The involvement of apoplastic invertase (Ap Inv) and sucrose synthase (SuSy) in the somatic embryo development of black spruce (Picea mariana) was investigated under different maturation conditions. Replacing 6% sucrose with 3% or 1% sucrose in the maturation medium drastically decreased Ap Inv activity and amount in embryogenic tissues. This was accompanied by a decrease in the hexose pool that resulted in a lower starch deposition and protein amount in embryogenic tissues together with a lower embryo production. Conversely, SuSy activity was stable during maturation regardless of the sucrose concentration used in the medium. The presence of an extracellular enzyme responsible for sucrose hydrolysis in the maturation medium was also verified. An immunodetection experiment with anti-acid invertase antibodies revealed the presence of an active 53 kDa polypeptide in the medium, which had a similar molecular mass to that of the Ap Inv polypeptide found in embryogenic tissues. Utilization of sucrose from the medium by the tissues was also studied using labelled 14C-sucrose. Distribution of the radioactivity between tissular sucrose, glucose, and fructose showed that sucrose was diffused into the cell wall of embryogenic tissues and partly hydrolyzed by Ap Inv. These results show that the utilization of sucrose from the medium, the Ap Inv activity in embryogenic tissues, and the release of an active invertase into the medium operate together for the utilization of the carbohydrates during somatic embryo development in black spruce.

Fructose↗

PubMed1989

The neural structures involved in cross-modal recognition and tactile discrimination performance: an investigation using 2-DG.

Given the failure of the ablation method to identify the neural structures/systems that are crucial for cross-modal recognition (CMR) and for tactile discrimination performance (TDP), we injected radioactive 2-deoxy-[14C]glucose (2-DG) into monkeys trained to a high level of CMR or TDP. Nine monkeys were trained to recognize in one sense-modality, to a level greater than 80% correct, the objects experienced in the alternate modality on only a single prior trial. After injection, CMR was continued (Expt. 1a) exactly as before, except that all CMR problems were now in only one direction; or (Expt. 1b) the second--the CMR--trial of each problem was not realized (i.e. no objects were available in the second modality) so that the monkey merely 'expected' the cross-modal trial. Nine other monkeys were trained on a graded roughness discrimination task with conventional 'titration' procedures to a stable level of performance, either (Expt. 2a) without having undergone any cortical removal, or (Expt. 2b) after unilateral removal--contralateral or ipsilateral to the preferred hand--of the posterior insula or of the second somatosensory projection cortex (SII). Exactly the same training procedures were continued in Expt. 2 after injection of 2-DG. Coronal radiographs were made at 490 microns for all 18 monkeys (and also for a 19th, which served as a control in Expt. 1b). The optical densities of the autoradiographs were measured quantitatively in respect of 50 structures or part-structures (e.g. sulci, thalamic nuclei, subcortical structures); they were rated either with the aid of 'pseudo-colours' produced by the computer, or directly by judging the black/white optical densities, in respect of 29 other structures. These 79 structures were then the dependent variables in MANOVAs or ANOVAs, to determine differences between groups; or within groups with respect to the left/right and ipsilateral/contralateral hemispheres; or interaction effects. It was found that certain structures (e.g. the ventral portion of the claustrum, the insula with its extension ventrally into the fundus, nucleus Medialis dorsalis and nucleus Pulvinar oralis) repeatedly, whereas others (e.g. the amygdala) never gave rise to significant outcomes. Structures in the left cerebral hemisphere were frequently found to be more strongly labelled than those in the right hemisphere.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1967

Occurrence and metabolism of scylloinositol in the locust.

1. scylloInositol has been identified as a component of locust haemolymph, where it occurs in concentrations of 0.2-0.4mg./ml. 2. A simple method for the identification of scylloinositol is described. This has been used to demonstrate the presence of scylloinositol in five other insect species. 3. Locust phospholipids contain myoinositol but no scylloinositol. 4. Radioactivity from [(14)C]glucose is incorporated into myoinositol and scylloinositol by the locust in vivo. 5. Extracts of locust fat body catalyse the conversion of myoinositol into scylloinositol. This seems to take place by a two-step process in which myoinositol is first oxidized with NAD(+) to myoinosose-2, and the myoinosose-2 is stereospecifically reduced with NADPH to scylloinositol.

Adipose Tissue↗

PubMed1968

Compartmentation between glycolysis and gluconeogenesis in rat liver.

1. The specific radioactivity-time relationships of glucose, glucose 6-phosphate, glycerol 1-phosphate and UDP-glucose were determined in rat liver after the intravenous injection of [U-(14)C]fructose, and a kinetic analysis was carried out. The glucose 6-phosphate pool was found to be compartmented into gluconeogenic and glycolytic components, and evidence was obtained that the triose phosphates were similarly compartmented. The glycolytic pathway was fed by glycogenolysis and glucose phosphorylation. There was no direct evidence that glycogenolysis fed only the glycolytic pathway, but this interpretation would make the liver resemble other organs in this respect. 2. UDP-glucose was not formed solely from gluconeogenic glucose 6-phosphate, as there was some dilution of label in the intervening glucose 1-phosphate pool, probably from glycogenolysis, though other pathways cannot be excluded. 3. The data cannot be explained by isotopic exchange.

Animals↗

PubMed1970

Measurement of uridine diphosphate glucuronic acid concentrations and synthesis in animal tissues.

1. A method for the isolation from animal tissues of UDP-glucuronic acid by one-dimensional paper chromatography is described and its concentrations in some tissues of several species of vertebrates are reported; the incorporation of [(32)P]-phosphate into UDP-glucuronic acid in vivo was also investigated. 2. The concentration of UDP-glucuronic acid was higher in the liver of rats, rabbits and guinea pigs than in the same tissue of some species of birds, amphibia and fishes; also, the concentration of UDP-glucuronic acid in rat liver, kidney and small intestine was several times lower than that of the same tissues of guinea pigs. 3. The rate of [(32)P]-phosphate incorporation into UDP-glucuronic acid was very high in rat liver and kidney and almost reached equilibrium with the radioactivity of UDP-glucose 30min after the administration of the [(32)P]phosphate.

Animals↗