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C Michoudet

Publications and source records attributed to C Michoudet.

11 recordsLinked to original sources
PubMed1994

Glutamine synthesis from glucose and ammonium chloride by guinea-pig kidney tubules.

1. At a physiological concentration (5 mM), glucose was found to be metabolized by isolated kidney cortex tubules prepared from fed guinea pigs. 2. The release of 14CO2 from [U-14C]glucose indicated that oxidation of the glucose carbon skeleton represented about 50% of the glucose removed; significant amounts of lactate and glutamine also accumulated. 3. Addition of 0.1-10 mM NH4Cl led to a dose-dependent stimulation of glucose metabolism which was accompanied by a large increase in lactate and glutamine accumulation and, to a lesser extent, in glucose oxidation. 4. Comparison of the release of 14CO2 from [1-14C]- and [6-14C]glucose indicates that, in both the absence and the presence of NH4Cl, the pentose phosphate shunt was only a minor pathway of glucose metabolism. 5. The central role of pyruvate carboxylase in the conversion of glucose carbon into glutamine carbon was demonstrated by using a bicarbonate-free medium and measuring the fixation of 14CO2 from [14C]bicarbonate, which was recovered mostly at C-1 of glutamine plus glutamate. 6. The NH4Cl-induced stimulation of glucose removal was secondary not only to increased glutamine synthesis, as shown by the effect of methionine sulphoximine, an inhibitor of glutamine synthetase, but also to the stimulation of phosphofructokinase activity by NH4Cl. 7. Renal arterio-venous difference measurements revealed that, in vivo, the guinea-pig kidney removed glucose from the circulating blood, which suggests that glucose carbon may contribute to the carbon skeleton of the glutamine released by this organ.

Ammonium Chloride↗
PubMed1992

Release and fixation of CO2 by guinea-pig kidney tubules metabolizing aspartate.

1. The metabolism of L-[U-14C]aspartate, L-[1-14C]aspartate and L-[4-14C]aspartate was studied in isolated guinea-pig kidney tubules. 2. Oxidation of C-1 plus that of C-4 of aspartate accounted for 90-92% of the CO2 released from aspartate, whereas oxidation of the inner carbon atoms of aspartate (which occurs beyond the 2-oxoglutarate dehydrogenase step) represented only 8-10% of aspartate carbon oxidation. 3. The formation of [1-14C]glutamine and [1-14C]glutamate from [1-14C]aspartate and [4-14C]aspartate indicated that about one-third of the oxaloacetate synthesized from aspartate underwent randomization at the level of fumarate. 4. With [U-14C]aspartate as substrate, the percentage of the C-1 of glutamate and glutamine found radiolabelled after 60 min of incubation was 92.7% and 47.5% in the absence and the presence of bicarbonate respectively. 5. That CO2 fixation occurred at high rates in the presence of bicarbonate was demonstrated by incubating tubules with aspartate plus [14C]bicarbonate; under this condition, the label fixed was found in C-1 of glutamate, glutamine and aspartate, as well as in C-4 of aspartate, demonstrating not only randomization of aspartate carbon but also aspartate resynthesis secondary to oxaloacetate cycling via phosphoenolpyruvate carboxykinase, pyruvate kinase and pyruvate carboxylase. 6. The importance of CO2 fixation in glutamine synthesis from aspartate is discussed in relation to the possible role of the guinea-pig kidney in systemic acid-base regulation in vivo.

Acetazolamide↗
PubMed1990

Glutamine synthesis from aspartate in guinea-pig renal cortex.

1. Glutamine was found to be the main carbon and nitrogen product of the metabolism of aspartate in isolated guinea-pig kidney-cortex tubules. Glutamate, ammonia and alanine were only minor products. 2. Carbon-balance calculations and the release of 14CO2 from [U-14C]aspartate indicate that oxidation of the aspartate carbon skeleton occurred. 3. A pathway involving aspartate aminotransferase, glutamate dehydrogenase, glutamine synthetase, phosphoenolpyruvate carboxykinase, pyruvate kinase, pyruvate dehydrogenase and enzymes of the tricarboxylic acid cycle is proposed for the conversion of aspartate into glutamine. 4. Evidence for this pathway was obtained by: (i) inhibiting aspartate removal by amino-oxyacetate, an inhibitor of transaminases, (ii) the use of methionine sulphoximine, an inhibitor of glutamine synthetase, which induced a large increase in ammonia release from aspartate, (iii) the use of quinolinate, an inhibitor of phosphoenolpyruvate carboxykinase, which inhibited glutamine synthesis from aspartate, (iv) the use of alpha-cyano-4-hydroxycinnamate, an inhibitor of the mitochondrial transport of pyruvate, which caused an accumulation of pyruvate from aspartate, and (v) the use of fluoroacetate, an inhibitor of aconitase, which inhibited glutamine synthesis with concomitant accumulation of citrate from aspartate.

Aminooxyacetic Acid↗
PubMed1989

Stimulation of glutamine metabolism by the antiepileptic drug, sodium valproate, in isolated dog kidney tubules.

The effects of sodium valproate, a widely used antiepileptic drug and an hyperammonemic agent, on glutamine and glutamate metabolism were studied in isolated dog kidney tubules. Valproate markedly stimulated glutamine removal as well as the formation of ammonia, aspartate, pyruvate, lactate, alanine and glucose; the increase in ammonia formation was explained by a stimulation by valproate of flux not only through glutaminase (EC 3.5.1.2) but also through glutamate dehydrogenase (EC 1.4.1.3). By contrast, valproate did not stimulate glutamate removal or ammonia, aspartate and glucose formation from glutamate; this suggests that the increase in flux through glutamate dehydrogenase with glutamine as substrate was secondary to the increase in flux through glutaminase. Accumulation of pyruvate, alanine and lactate in the presence of valproate was much less from glutamate than from glutamine. Inhibition by amino-oxyacetate of accumulation of aspartate and alanine from glutamine caused by valproate did not prevent the acceleration of glutamine utilization and the subsequent stimulation of ammonia formation. These data are consistent with a stimulatory effect of valproate primarily exerted at the level of glutaminase in dog kidney tubules. However, the fact that assayed activity of glutaminase remained unchanged in the presence of valproate suggests that this compound accelerates flux through the latter enzyme by an indirect mechanism probably related to the renal metabolism of this compound.

Alanine↗
PubMed1988

Pyruvate carboxylation in glutamine synthesis from alanine by isolated guinea-pig renal cortical tubules.

Isolated guinea-pig kidney cortex tubules were incubated in Krebs-Henseleit buffer containing NaH14CO3 (25 mM) and L-alanine (5 mM). A high rate of alanine metabolism was found to be accompanied by a high rate of both 14CO2 fixation and glutamine synthesis. The fixation of 14CO2 was virtually abolished in the presence of oxalate, a known inhibitor of pyruvate carboxylase, indicating that, in guinea-pig renal cortex, this enzyme is responsible for the synthesis of oxaloacetate in the conversion of alanine into glutamine. More than 90% of the label fixed was found in carbon 1 mainly of glutamine and to a lesser extent of glutamate. In the presence of alanine + NaH14CO3 + MSO, an inhibitor of glutamine synthetase, most of the 14CO2 fixed by pyruvate carboxylase was subsequently released and carbon 1 of glutamate was the only site of labelling. In the presence of alanine + NaH14CO3, the fact that not all the glutamine found was labelled in carbon 1 could be explained by glutamine synthesis from endogenous substrates as well as by glutamine synthesis from alanine after prior equilibration of [4-14C]-oxaloacetate with fumarate; that such equilibration occurred was demonstrated by the observation that [1-14C]-glutamine and [1-14C]-glutamate were synthesized from [1-14C]-alanine.

Alanine↗
PubMed1987

Characteristics of acetaldehyde metabolism in isolated dog, rat and guinea-pig kidney tubules.

The metabolism of acetaldehyde was studied in isolated dog, rat and guinea-pig kidney-cortex tubules. In contrast with previous observations of Cederbaum and Rubin in rat kidney mitochondria (Archs Biochem. Biophys. 179, 46-66 1977) acetaldehyde was found to be metabolized by the tubules at high rates and in a dose-dependent manner at concentrations up to 5-10 mM. At high acetaldehyde concentrations (1-10 mM) acetaldehyde removal was accompanied by a high rate of acetate accumulation which explained most of the acetaldehyde metabolized in dog and guinea-pig but not in rat kidney tubules. These species differences in acetaldehyde metabolism can be explained by the differences in activities of aldehyde dehydrogenase (EC 1.2.1.3) and acetyl-CoA synthetase (EC6.2.1.1), the enzymes involved in renal acetaldehyde metabolism which were measured in the renal cortex of the three species. The acetaldehyde carbon removed and not accounted for by acetate accumulation was completely oxidized to CO2 as demonstrated by the measurement of [U-14C]-acetaldehyde conversion into 14CO2. At "physiological" acetaldehyde concentrations (0.1 and 0.2 mM) acetaldehyde utilization was also concentration-dependent but no acetate accumulation was observed.

Acetaldehyde↗
PubMed1987

Metabolism of acetaldehyde in human and baboon renal cortex. Ethanol synthesis by isolated baboon kidney-cortex tubules.

Acetaldehyde (1-20 mM) was metabolized at high rates and in a dose-dependent manner in isolated human and baboon kidney-cortex tubules. Acetaldehyde removal was accompanied by a large accumulation of acetate in both human and baboon tubules. By contrast, a large synthesis of ethanol was observed only in baboon tubules. Consistent with the latter finding, ethanol was found to be metabolized at significant rates in baboon but not human tubules. In the tubules from both species, a significant fraction of the acetaldehyde removed was also completely oxidized to CO2 and H2O. These results suggest that, in both man and baboon, the kidneys participate in the in vivo metabolism of acetaldehyde; they also suggest that, in contrast with the human kidneys, the baboon kidneys contribute to the detoxication of circulating ethanol.

Acetaldehyde↗
PubMed1987

The choice of the wavelengths in spectral analysis: an application to determine the composition of a tetraribonucleotide mixture.

We present a simple program running on a pocket computer allowing us to determine the order of importance of the wavelengths (regarding the accuracy of the results) in spectral analysis and to evaluate the absolute errors made on the determination of the concentrations of the constituents. We have applied this programme to the determination of the composition of a tetraribonucleotide mixture and have compared our results with other results published.

Microcomputers↗
PubMed1985

Quantitative technique for bio-electrical spectroscopy.

Bio-electrical impedance measurements have been widely used for the study of body tissues. Apart from recordings of biophysical signals (respiration, perfusion, cardiac output, red cell settling, etc.) measurements of specific resistivity of a tissue provide information about its pathological state. Mapping electrical parameters will give more detailed information. The interpretation of recorded data and the design of equipment, both require a preliminary knowledge of the values encountered under normal and pathological conditions. The purpose of the technique described here is to determine the complex resistivity of breast tissue samples in vitro at frequencies between 0.5 kHz and 1 MHz. The equipment is described and the calibration procedure explained. A calculation of the final error interval is given. Characteristic spectra of modulus and phase angle recorded in normal and pathological breast tissue are shown. The technique can however be used for other body tissues, typical applications being fundamental tissue studies and a determination of the most suitable frequencies for use with impedance measuring devices. In clinical practice it could contribute to the determination of intra- and extracellular volume, the monitoring of transplanted organs and the examination of surgically treated tumours, and to any technique based on tissue characterization.

Biomedical Engineering↗