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PubMed1982

Mapping of the neural activity in the cerebellum of the mouse during stepping by means of 2-deoxyglucose autoradiography.

Incorporation of radioactive 2-deoxy-D-glucose (2-DG) into the cerebellum of mice which were resting in a cage or stepping on a treadmill was examined by means of autoradiography. In the cerebellum of the mouse resting in the cage, a large amount of 2-DG was incorporated into the posterior vermis, the flocculus, cerebellar nuclei and vestibular nuclei. On the other hand, during stepping, incorporation of 2-DG was increased remarkably in the anterior vermis, the pyramis, the anterior part of the ansiform lobule and the paraflcculus, indicating that these regions are functionally involved in locomotor movements.

Animals↗
PubMed1989

Synthesis of glutamate and glutamine in dibutyryl cyclic AMP-treated astrocytes.

The relative contributions of radioactively labeled fatty acids and glucose to synthesis of glutamate and glutamine were compared in native and dibutyryl cyclic AMP (diBcAMP)-treated primary rat astrocytes. The intracellular specific activities of glutamate and glutamine were 10-fold greater than the specific activities of aspartate or alanine. Butyrate, octanoate and palmitate were equally as effective as precursors for glutamate and glutamine while glucose was 50% as effective as the fatty acids. The specific activity of glutamate and glutamine were identical in the absence of diBcAMP. In diBcAMP treated cells the specific activity of glutamine was greater than that of glutamate when octanoate and palmitate were the labeled precursors. This suggests that cultured astrocytes preferentially utilize free fatty acids for glutamate/glutamine synthesis and that diBcAMP-treated astrocytes contain more than one glutamate compartment.

Animals↗
PubMed1975

The effect of methionine on the metabolism of serine and glycine in vitamin B-12-deficient rats.

The effect of methionine supplementation on glycine and serine metabolism was studied in vitamin B-12-deficient rats which received only 0.2% methionine in the diet. In the perfused liver, incorporation of the C-2 of glycine to the C-3 of serine was increased by addition of methionine to the perfusate. The oxidation of [1-14C]glycine to 14CO2 was however depressed. Unlike methionine, glycine did not have any significant effect on the liver folate coenzyme distribution. Oxidation of [3-14C]serine to 14CO2 both in vivo and in perfused liver was increased by methionine. A major portion of the C-3 radioactivity however was recovered in glucose. Data presented indicate that the rate of oxidation of [2-14C]histidine to 14CO2 is a more sensitive indicator of folate deficiency than the rate of oxidation of [3-14C]serine to 14CO2 although both are presumably tetrahydrofolate dependent.

Animals↗
PubMed1992

Effect of inorganic mercury on in vitro placental nutrient transfer and oxygen consumption.

The effect of mercury (HgCl2) on placental amino acid and glucose transfer as determined by the use of their nonmetabolizable radioactive analogues, aminoisobutyric acid (AIB) and 3-o-methyl glucose (3MG), respectively, was studied in an in vitro perfusion model of a term human placenta. Hg2+ was found to decrease the transfer and accumulation of AIB without affecting 3MG transfer. It was also found to decrease the placental oxygen consumption rate. Placental circulation and tissue morphology remained intact, as demonstrated by the antipyrine transfer rate, and by electron microscopy, respectively. The mechanism by which Hg2+ may interfere with placental amino acid transfer and accumulation is discussed. Although much higher concentrations than those found in the ordinary polluted environment were used, this is the first report showing that Hg2+ interferes with an essential human placental function in a system employing a whole human placental cotyledon. This finding may indicate the possible involvement of Hg2+ in impaired organogenesis in early pregnancy or deranged fetal growth during the last trimester.

3-O-Methylglucose↗
PubMed1983

Metabolism of leucine by the isolated perfused goat udder.

Seven lactating goat mammary glands from 6 goats were perfused for several hours in the presence of [U-14C]L-leucine (4 experiments) or [2-3H; 1-14C]DL-leucine (3 experiments) and received adequate quantities of glucose, acetate and amino acids. Radioactivity in casein was mainly recovered in leucine and 90% of casein leucine was derived from free plasma leucine. About 64% of the leucine molecules were used for casein synthesis. Up to 12% of the molecules were channelled into lipid synthesis, while the remaining (up to 24%) were metabolized to CO2. From the 3H/14C ratio of casein and casein leucine, it was calculated that 70-80% of the leucine molecules were reversibly transaminated before their incorporation into milk protein. However, only 4-8% of the plasma leucine molecules were transaminated during passage through the udder. Different pools for oxidation and for protein synthesis may be present in the goat mammary gland.

Amino Acids↗
PubMed1971

Lipid metabolism by rat lung in vitro. Effect of starvation and re-feeding on utilization of (U- 14 C)glucose by lung slices.

1. The incorporation of [U-(14)C]glucose into several lipid components of lung and liver slices, and the activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), ;malic' enzyme (EC 1.1.1.40) and NADP-isocitrate dehydrogenase (EC 1.1.1.42) of the cell cytosol were examined in normal, starved and re-fed rats. 2. Lipogenesis and the activities of these enzymes in liver were decreased markedly in rats starved for 72h. Re-feeding starved rats on a fat-free diet for 72h resulted in the well documented hyperlipogenic response in liver, particularly in its ability to convert glucose into neutral lipid, and increased activities of glucose 6-phosphate dehydrogenase, ;malic' enzyme and 6-phosphogluconate dehydrogenase to values approx. 700, 470 and 250% of controls respectively. 3. Approx. 70% of the total label in lung lipids was present in the phospholipid fraction. Hydrolysis of lung phospholipids revealed that lipogenesis from glucose was considerable, with approx. 40% of the total phospholipid radioactivity present in the fatty acid fraction. 4. Incorporation of glucose into total lung lipids was decreased by approx. 40% in lung slices of starved rats and was returned to control values on re-feeding. Although phospholipid synthesis from glucose was decreased in lung slices of starved rats, the decrease proportionally was greater for the fatty acid fraction (approx. 50%) as compared with the glycerol fraction (approx. 25%). 5. The activities of lung glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and NADP-isocitrate dehydrogenase were not affected by the dietary alterations. ;Malic' enzyme activity was not detected in lung cytosol preparations. 6. The results are discussed in relation to the surface-active lining layer (surfactant) of the lung.

Animals↗
PubMed1971

Kinetin and carbohydrate metabolism in chinese cabbage.

The effects of kinetin on starch and sugar levels and on (14)CO(2) and (32)P-orthophosphate labeling patterns of floated Chinese cabbage (Brassica pekinensis) leaf discs were investigated. Kinetin caused gross starch degradation. Neutral sugars were depressed by 30 to 40% in leaf tissue treated with kinetin for 24 hours. (14)CO(2) labeling of leaf discs pretreated with kinetin for 24 hours showed increased radioactivity in chloroform-soluble material and most sugar phosphates, and a 35 to 40% decrease in radioactivity in the neutral sugars, glucose, sucrose, and fructose. Incorporation into ATP was increased by 40% by kinetin. (32)P-Orthophosphate uptake was inhibited 30% by kinetin. When corrected for uptake, kinetin stimulated incorporation into chloroform-soluble material but had little effect on other cell fractions. These results indicate that kinetin mobilizes starch reserves and increases the flow of sugars required for the synthesis of lipids and structural materials in floated discs.

Journal Article↗
PubMed1977

The hypoglycaemic action of somatostatin in the anaesthetized dog.

The hypoglycaemic action of somatostatin was investigated in fasted anaesthetized dogs. An isotopic turnover technique with radioactive D-[2-3H] glucose tracer was used to measure the rates of hepatic production and peripheral utilization of glucose before, during, and after a 60min infusion of 150 mug somatostatin. Plasma concentrations of insulin, glucagon, growth hormone, cortisol and free fatty acids were also measured. Somatostatin infusion caused a fall of plasma glucose demonstrable within 10 min. After 60 min plasma glucose had fallen to 75% of the pre-infusion concentration. Simultaneoulsy the specific activity of the D-[2-3H] glucose increased and calculation of the rates of production and utilization of glucose over 10 min intervals indicated that the mean hepatic production rate of glucose fell to 59% of the pre-infusion rate while the rate of peripheral utilization of glucose fell to 83% of the pre-infusion rate. These results show that the hypoglycaemic action of intravenous somatostatin depends exclusively upon the inhibition of hepatic glucose production. This effect may have been partly due to a fall of plasma glucagon concentration and occurred despite a fall of plasma insulin to less than 1 mu 1(-1).

Animals↗
PubMed1974

Lipid peroxidation in the killing of phagocytized pneumococci.

To directly examine the role of hydrogen peroxide in the killing of bacteria after ingestion by granulocytes, we have studied some of the events of phagocytosis of a mutant strain of pneumococci which is relatively deficient in peroxide production. The hydrogen peroxide-deficient pneumococci and the otherwise identical wild type were grown with [(14)C]arachidonic and [(3)H]palmitic acid labels to label their lipids with unsaturated and saturated fatty acids, respectively. They were then incubated with both normal and chronic granulomatous disease granulocytes. The rates of ingestion and bacterial killing and the stability of fatty acids in the cell-bacteria complex were followed. Radioactive carbon dioxide released from glucose was also independently followed to measure glucose oxidation. Ingestion was similar for all cell-bacteria combinations. Chronic granulomatous disease cells killed the peroxide-positive wild pneumococci much more effectively (20-fold) than the peroxide-deficient mutant. Normal cells killed both peroxide-positive and -negative strains effectively. A considerable loss of [(14)C]arachidonic acid ( approximately 40%) consistent with lipid peroxidation of this unsaturated fatty acid was observed in all normal cells and in chronic granulomatous disease cells with peroxide-positive pneumococci. However, no loss of [(14)C]arachidonic acid occurred in chronic granulomatous disease cells with the peroxide-deficient pneumococci. No loss tritiated palmitic acid occurred in any cell-bacteria combination. Glucose oxidation was impaired in the chronic granulomatous disease cells in comparison to normal cells at rest and was especially impaired in chronic granulomatous disease cells ingesting the peroxide-deficient mutant pneumococci. This defect was partially corrected after phagocytosis of the peroxide-positive strain. These data directly support the hypothesis that bacterial killing is partially dependent upon an intact peroxide-generating system in the leukocyte-bacteria complex. Moreover, they indicate that bacterial lipid peroxidation is associated with the generation of peroxide during phagocytosis. Finally, they suggest that such peroxidation may contribute to effective phagocytic bacterial killing.

Animals↗
PubMed1977

Rate of ribosomal ribonucleic acid chain elongation in Escherichia coli B/r during chloramphenicol treatment.

In Escherichia coli B/r growing in glucose-amino acids medium, the radioactive labeling of 5S ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) was measured after the simultaneous addition to the bacteria of chloramphenicol (CAM) (100 mug/ml), rifampin (200 mug/ml), and radioactive uracil. Accumulation of 5S rRNA ceased 85 s after the addition of rifampin, independent of the presence or absence of CAM; this indicates that CAM did not affect the rRNA chain growth rate. Together with previous measurements of the synthesis of rRNA and messenger RNA under these conditions, the results imply that CAM caused a redistribution of RNA polymerase which greatly favored stable RNA synthesis (77 to 97% of total functioning RNA polymerase engaged in synthesis of rRNA and tRNA). Further, it is inferred that RNA polymerase molecules were activated that were inactive during exponential growth. The labeling of tRNA observed under these conditions suggests the existence of clusters of tRNA genes at the 3' end of long transcripts that resemble the rRNA precursor in length and response to CAM and may be parts of rRNA transcripts.

Chloramphenicol↗
PubMed1978

Nucleotides and coenzymes in nuclei isolated from rat liver.

In rat-liver nuclei, isolated by the non-aqueous technique, the concentrations and labelling rates of the purine moiety of acid-soluble nucleotides were determined and compared with corresponding data for non-fractionated tissue and nuclei-free cytoplasm. Livers were used from untreated rats, from rats with a highly stimulated synthesis of NAD and from rats following a heavy metabolic load with adenosine. Under all circumstances, the nuclear and cytoplasmic concentrations of nucleotides (e.g. ATP and its dephosphorylated forms, pyridine nucleotides) and of free glucose were practically identical. Specific radioactivities after a pulse with formate also indicated a nucleo-cytoplasmic equilibrium for purine-containing nucleotides. It is concluded that precursor pools for nuclear biosyntheses as well as energy supply for other nuclear activities may be determined by an analysis of the non-fractionated tissue.

Adenine Nucleotides↗
PubMed1982

Estimation of free myo-inositol in milks of various species and its source in milk of rats (Rattus norvegicus).

Free myo-inositol content of milk of rats and other rodents fed standard laboratory chow is 80 mg/100 g as compared to about 4 mg/100 g for cow milk. Feeding experiments with graded amounts of myo-inositol demonstrated that the myo-inositol content of milk is strongly influenced by dietary intake. Radioactivity from either [carbon-14] glucose or [hydrogen-3] myo-inositol injected intraperitoneally was incorporated rapidly into milk myo-inositol. Thus, both mammary biosynthesis and active transport contribute to the high myo-inositol in rat milk, but the latter is quantitatively the more important.

Animals↗
PubMedDate not supplied

[The significance of the structure of dietary oligosaccharides in their metabolism into liver lipids].

In experiments on 24 Wistar male rats the time course of incorporation of radioactive carbon and hydrogen of glucose and fructose into liver lipids was studied, as well as the influence on this process of the animals feeding with rations containing sucrose and inverted sugar (17% calory value). A mathematical model has been suggested for the evaluation of the effect of sugar and a glucose-fructose mixture on monosugar utilization into liver lipids. It was found that sucrose led to an increase in the rate of the label incorporation into liver lipids from glucose and fructose, and to a growth in the constant of the label release from lipids, i.e. induced acceleration of the liver lipid turnover. These effects were absent in rats given inverted sugar.

Animals↗
PubMed1988

[Biochemical study on nonenzymatic glycosylation of human erythrocyte Cu-Zn-superoxide dismutase].

Human erythrocytes contain glycosylated and nonglycosylated Cu-Zn-superoxide dismutases which can be separated by boronate affinity chromatography. The percentage of the glycosylated form is significantly increased in the erythrocytes of the patients with diabetes as compared to normal erythrocytes. The nonglycosylated Cu-Zn-superoxide dismutase, which was washed through the boronate column, was glycosylated in vitro upon exposure to radioactive or nonradioactive D-glucose. Incorporation of D-glucose into the protein was observed, and with increase in glycosylation, the enzymatic activity decreased, indicating that the glycosylation of the enzyme led to low active form. The sites of glycosylation of the superoxide dismutase were identified by amino acid analysis after reverse-phase high performance liquid chromatography of the trypsin-treated peptides.

Chromatography, Affinity↗
PubMedDate not supplied

[Glycogen and fat deposition from labelled precursors at high levels of sugar consumption].

The feeding of rats during two weeks with a high-carbohydrate diet, in which sucrose presented 40% of energy, induced an increased inclusion of the radioactive label from 3H-glucose and 14C-acetate into store lipids, and 3H-glucose inclusion into the liver and muscle glycogen. The ratio 3H/14C in glycogen and store lipids after administration of the mixture of 3H-glucose and 14C-acetate to the animals reflected a specific effect of sucrose in high-carbohydrate diets.

Adipose Tissue↗
PubMed1975

The response of red cell hexose monophosphate shunt after sulfhydryl inhibition.

In this investigation, we studied the importance of cellular glutathione (GSH) in the hexose monophosphate shunt (HMPS) activity of unstimulated human erythrocytes and the mechanism by which pyruvate stimulates the HMPS. The rate of HMPS activity was measured by the production of radioactive CO2 from 14C-1-glucose or 14C-1-ribose using a vibrating reed electrometer and ionization chamber. HMPS activity was not significantly impaired by N-ethylmaleimide (NEM) in concentrations which bound all red cell GSH. Red cells incubated under carbon monoxide (CO), an experimental condition which eliminates peroxide production, still had HMPS activity which was 44% of the value under air. Pyruvate stimulation of the HMPS was unaffected by doses of NEM which bound all cellular GSH or by incubation under CO. These data indicated that pyruvate stimulation of the HMPS occurs by pathways which do not involve peroxide formation, GSH, or oxygen. This study indicates that sulfhydryl blockade of GSH does not necessarily inhibit HMPS activity and that HMPS activity in red cells may respond to reactions not linked directly to glutathione reduction.

Blood Glucose↗
PubMed1967

Measurement of flow of carbon atoms from glucose and glycogen glucose to glyceride glycerol and glycerol in rat heart and epididymal adipose tissue. Effects of insulin, adrenaline and alloxan-diabetes.

1. Flow of carbon atoms from glucose and glycogen glucose to glyceride glycerol, glyceride fatty acids and glycerol was calculated in the perfused rat heart and incubated epididymal adipose tissue from the incorporation of (14)C from [U-(14)C]-glucose (into glyceride glycerol, glyceride fatty acids and glycerol in the medium), and from measurements of the specific activity of l-glycerol 3-phosphate, and the effects of insulin, adrenaline and alloxan-diabetes were studied. Measurements were also made of the uptake of glucose and the outputs of lactate, pyruvate and glycerol. 2. New methods are described for the measurement of radioactivity in small amounts of metabolites (glycerol, glucose 6-phosphate and fructose 6-phosphate and l-glycerol 3-phosphate) in which use has been made of alterations in charge induced by enzymic conversions to effect resolution by ion-exchange chromatography. 3. In hearts the specific activity of l-glycerol 3-phosphate was less than that of glucose in the medium but similar to that of lactate released during perfusion. Because repeated measurements of the specific activity of l-glycerol 3-phosphate was impracticable, the specific activity of lactate has been used as an indirect measurement of glycerol phosphate specific activity. 4. In fat pads, specific activity of lactate was the same as that of glucose in the medium and thus the specific activity of l-glycerol 3-phosphate was taken to be the same as that of medium glucose. 5. In hearts from alloxan-diabetic rats, despite decreased glucose uptake and l-glycerol 3-phosphate concentration, flow of carbon atoms through l-glycerol 3-phosphate to glyceride glycerol was increased about threefold. 6. In fat pads, flow of carbon atoms through l-glycerol 3-phosphate to glyceride glycerol was increased by insulin (twofold), by adrenaline in the presence of insulin (fivefold) and by diabetes in pads incubated with insulin (1.5-fold). These increases could not be correlated either with increases in glucose uptake, which was unchanged by adrenaline and decreased in diabetes, or with the concentration of l-glycerol 3-phosphate, which was decreased by adrenaline and unchanged in diabetes. 7. These results are discussed in relation to the control of glyceride synthesis in heart and adipose tissue and to the regulation of glyceride fatty acid oxidation in the perfused rat heart.

Adipose Tissue↗