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R Parrilla

Publications and source records attributed to R Parrilla.

At least 73 records · Page 4Linked to original sources
PubMed1986

Pyruvate decarboxylating action of L-cycloserine. The significance of this in understanding its metabolic inhibitory action.

We present evidence which demonstrates that L-cycloserine, structural analog of L-alanine, which is known to be an effective aminotransferase inhibitor, is also a potent inhibitor of cellular pyruvate metabolism. This effect was found to be related to its almost instantaneous action in decreasing pyruvate concentrations in a dose-dependent manner. 1H nuclear magnetic resonance studies clearly demonstrate that the irreversible removal of pyruvate induced by L-cycloserine is caused by the decarboxylating action of the latter. Pyruvate disappearance induced by L-cycloserine can be stoichiometrically accounted for as acetate. The process does not involve any chemically detected transformation of L-cycloserine. These observations lead to two main considerations regarding the known action of L-cycloserine. First, its inhibitory effect on gluconeogenesis from lactate could be explained only on the basis of its ability to reduce pyruvate availability with no apparent need for transaminase inhibition. Second, its ability as a transaminase inhibitor should be reconsidered in view of its potent decarboxylating action on pyruvate and probably other oxoacids.

Animals↗
PubMed1986

Rate-limiting steps for hepatic gluconeogenesis. Mechanism of oxamate inhibition of mitochondrial pyruvate metabolism.

Oxamate, structural analog of pyruvate, inhibits gluconeogenesis from pyruvate or substrates yielding pyruvate. The inhibitory effect is the result of a decreased mitochondrial pyruvate utilization. Although the inhibition of gluconeogenesis is competitive for pyruvate, in isolated mitochondria oxamate displays a mixed type kinetics inhibitory pattern of pyruvate utilization. Evidence is presented indicating that this mixed type pattern of inhibition is the result of the action of oxamate on two different sites: noncompetitive inhibition of pyruvate carboxylation, and competitive inhibition of pyruvate entry into the mitochondria. At concentrations of pyruvate above 0.4 mM, although pyruvate carboxylation is decreased by 40% by oxamate, no detectable effects on the gluconeogenic flux were observed. This finding strongly indicates that pyruvate carboxylase is not an important rate-limiting step for hepatic gluconeogenesis. Thus, the inhibition of gluconeogenesis at low pyruvate concentrations (less than 0.4 mM) seems to be the result of an interaction of oxamate with the mitochondrial pyruvate translocator, indicating that pyruvate transport across the mitochondrial membrane is the first nonequilibrium step in the gluconeogenic pathway when low physiological concentrations of this substrate are utilized.

Alanine↗
PubMed1986

Interrelation between gluconeogenesis and hepatic protein synthesis.

Acute administration of glucagon to the rat in vivo inhibits hepatic polypeptide chain elongation by about 30%. This effect was not observed in adrenalectomized rats, despite the significant increases in the hepatic content of cyclic AMP. Fatty acid administration mimics the glucagon action on protein synthesis; however, in adrenalectomized animals they were ineffective. Whether glucagon or fatty acids were administered, there was a significant increase in the state of reduction of the NAD system in normal as well as in adrenalectomized rats. This observation rules out the change in the cellular state of reduction as the mediator of their action on protein synthesis. A correlation was observed between the ability of glucagon or fatty acids to inhibit protein synthesis and to stimulate gluconeogenesis. An increased biosynthetic activity as reflected by an increased gluconeogenic flux is accompanied by a decreased phosphorylation state of adenine nucleotides that might be responsible for the inhibitory effect on protein synthesis. In adrenalectomized animals in which neither glucagon nor fatty acids stimulate gluconeogenesis, no effects on phosphorylation state or on the rate of protein synthesis were detected.

Adenine Nucleotides↗
PubMed1986

Interaction of oxamate with the gluconeogenic pathway in rat liver.

Oxamate, a structural analog of pyruvate, known as a potent inhibitor of lactic dehydrogenase, lactic dehydrogenase, produces an inhibition of gluconeogenic flux in isolated perfused rat liver or hepatocyte suspensions from low concentrations of pyruvate (less than 0.5 mM) or substrates yielding pyruvate. The following observations indicate that oxamate inhibits flux through pyruvate carboxylase: accumulation of substrates and decreased concentration of all metabolic intermediates beyond pyruvate; decreased levels of aspartate, glutamate, and alanine; and enhanced ketone body production, which is a sensitive indicator of decreased mitochondrial free oxaloacetate levels. The decreased pyruvate carboxylase flux does not seem to be the result of a direct inhibitory action of oxamate on this enzyme but is secondary to a decreased rate of pyruvate entry into the mitochondria. This assumption is based on the following observations: Above 0.4 mM pyruvate, no significant inhibitory effect of oxamate on gluconeogenesis was observed. The competitive nature of oxamate inhibition is in conflict with its effect on isolated pyruvate carboxylase which is noncompetitive for pyruvate. Fatty acid oxidation was effective in stimulating gluconeogenesis in the presence of oxamate only at concentrations of pyruvate above 0.4 mM. Since only at low pyruvate concentrations its entry into the mitochondria occurs via the monocarboxylate translocator, from these observations it follows that pyruvate transport across the mitochondrial membrane, and not its carboxylation, is the first nonequilibrium step in the gluconeogenic pathway. In the presence of oxamate, fatty acid oxidation inhibited gluconeogenesis from lactate, alanine, and low pyruvate concentrations (less than 0.5 mM), and the rate of transfer of reducing equivalents to the cytosol was significantly decreased. Whether fatty acids stimulate or inhibit gluconeogenesis appears to correlate with the rate of flux through pyruvate carboxylase which ultimately seems to rely on pyruvate availability. Unless adequate rates of oxaloacetate formation are maintained, the shift of the mitochondrial NAD couple to a more reduced state during fatty acid oxidation seems to decrease mitochondrial oxaloacetate resulting in a decreased rate of transfer of carbon and reducing power to the cytosol.

Alanine↗
PubMed1986

Effect of ethanol on proteolysis in isolated liver cells.

Ethanol, when tested alone, inhibited proteolysis by about 20%; however, no effect was detected when it was combined with exogenous oxidizable fuels which inhibited proteolysis by themselves. Ethanol was effective in inhibiting proteolysis in the presence of protease inhibitors like ammonia, leupeptin or methylamine, indicating that its mechanism of action involves a non-lysosomal pathway of degradation. Ethanol oxidation is mandatory for it to have effect on proteolysis, however, its action is not related to an increased state of reduction of the NAD system. In contrast to other reductants of the NAD system, ethanol effect is accompanied by a rise in the phosphorylation state of the adenine nucleotides, suggesting that its action might be related to the cellular energy state.

Alcohols↗
PubMed1985

1H NMR detection of cerebral myo-inositol.

A previously unassigned group of prominent multiplets of the 360 MHz 1H NMR spectrum of acid stable metabolite extracts from rat brain is shown to arise from free myo-inositol. This conclusion is derived from a systematic analysis of the high-resolution 1H NMR spectra of brain acid extracts, in which appropriate conditions and optimal proton signals have been selected for the quantitative analysis of up to 15 metabolites. Developmental variations in the cerebral content of myo-inositol could be readily detected using this approach, which provides a novel alternative to study myo-inositol metabolism under physiological or pathological conditions.

Animals↗
PubMed1983

Fatty acid-glucose interaction in the control of protein synthesis by isolated rat lung cells.

(1) The addition of long chain fatty acids to the incubation medium of isolated rat lung cells produced a dose-dependent inhibition of protein labelling from L-[3H]valine. Maximal rate changes were observed at fatty acids levels within the range of their physiological concentration. (2) The effect of fatty acids on protein labelling does not seem to be mediated by their oxidation. The following observations seem to support this conclusion: (a) the rate of fatty acid oxidation by lung cells was remarkably low, so that no significant variations in the state of reduction of the NAD system were detected; (b) there was no correspondence in the dose-response patterns of fatty acid oxidation and inhibition of protein labelling; (c) octanoate was much more actively oxidized than oleate, however the latter was more effective in decreasing protein labelling. (3) An apparent relationship between the length of the fatty chain and its ability to inhibit protein labelling seems to exist. The longer the chain the stronger the inhibitory effect observed. (4) The effect of fatty acid on protein labelling seems to be mediated by a cellular energy depletion secondary to an inhibition of the respiratory chain. Their ability to decrease oxygen uptake and adenine nucleotide content was also proportional to the chain length. (5) Glucose, which apparently acted by increasing energy production at substrate level phosphorylation, partially prevented the inhibitory effect of fatty acid on protein labelling. This observation supports the point of view that fatty acids do not act in decreasing protein labelling by perturbing directly the protein synthesis machinery but decreasing the phosphorylation potential.

Adenine Nucleotides↗
PubMed1983

Acute effects of ethanol in the control of protein synthesis in isolated rat liver cells.

The acute effect of ethanol on hepatic protein synthesis is a rather controversial issue. In view of the conflicting reports on this subject, the effect of ethanol on protein labeling from L-[3H]valine in isolated liver cells was studied under a variety of experimental conditions. When tracer doses of the isotope were utilized, ethanol consistently decreased the rate of protein labeling, regardless of the metabolic conditions of the cells. This inhibition was not prevented by doses of 4-methylpyrazole large enough to abolish all the characteristic metabolic effects of ethanol, and it was not related to perturbations on the rates of L-valine transport and/or proteolysis. When ethanol was tested in the presence of saturating doses of L-[3H]valine no effect on protein labeling was observed. These observations suggest that the ethanol effect in decreasing protein labelling from tracer doses of the radioactive precursor does not reflect variations in the rate of protein synthesis but reflects changes in the specific activity of the precursor. These changes probably are secondary to variations in the dimensions of the amino acid pool utilized for protein synthesis. Even though it showed a lack of effect when tested alone, in the presence of saturating doses of the radioactive precursor ethanol inhibited the stimulatory effects on protein synthesis mediated by glucose and several gluconeogenic substrates. This effect of ethanol was not prevented by inhibitors of alcohol dehydrogenase, indicating that a shift of the NAD system to a more reduced state is not the mediator of its action. It is suggested that ethanol probably acted by changing the steady-state levels of some common effector(s) generated from the metabolism of all these fuels or else by preventing the inactivation of a translational repressor.

Animals↗
PubMed1982

Role of the adenylate system and glycolytic flux in the control of protein synthesis in isolated rat lung cells.

(1) Glucose stimulates the incorporation of amino acids into protein in lung cells isolated by digestion of the lung stroma with collagenase. This effect reflects mainly an increase in protein synthesis since no effect of glucose had been found to the uptake of amino acid precursors and, although glucose decreases the rate of intracellular proteolysis by 15%, this effect cannot account for the increased incorporation of radioactivity into proteins. Furthermore, glucose did not induce any significant change in the intracellular content of valine. (2) For glucose to act on protein synthesis, it must be glycolyzed since its stereoisomer, L-glucose, which is not metabolized by lung cells, has no effect. (3) The mechanism of glucose action does not seem to be related simply to variations of cellular ATP content or energy charge. The following arguments seem to support this conclusion: (i) glucose does not bring about significant variations in the concentration of reactants of the adenylate system; (ii) the increase in protein synthesis induced by glucose in energy-depleted cells correlates with a rise in ATP content and energy charge; however, adenosine, which increases ATP levels in a form quantitatively similar to glucose, is unable to affect protein synthesis: (iii) glucose also accelerates the incorporation of amino acids into proteins in adenosine-treated lung cells in which the ATP concentration was almost double that of the control and the energy charge was considerably elevated, ruling out the possibility that a rise in the steady-state concentration of ATP and/or energy charge alone could be responsible for the acceleration of protein synthesis. (4) It can be concluded that the effect of glucose in increasing protein synthesis in lung cells is dependent on some signal arising from its breakdown and not to variations in the concentration of reactants or energy charge of the adenylate system.

Adenine Nucleotides↗
PubMed1982

Role of the state of reduction of the NAD system on the regulation of hepatic protein synthesis in the rat in vivo.

1. The administration of octanoate to rats in vivo increased the state of reduction of the hepatic NAD system and decreased the phosphorylation potential. This effect was accompanied by a 20% inhibition of protein synthesis. 2. The acute administration of ethanol produced similar reduction of the hepatic NAD system; however, in contrast to octanoate no effect on the phosphorylation potential was detected and rates of protein synthesis were unaffected. 3. It is concluded that a rise in the state of reduction of the NAD system is not effective in decreasing hepatic protein synthesis in vivo unless it is accompanied by a decrease in the phosphorylation potential.

Adenosine Diphosphate↗
PubMed1978

Flux of metabolic fuels during starvation in the rat.

Rats were starved for 6 days to determine whether the "nitrogen sparing" observed during starvation in humans was also present in rats. The urinary nitrogen excretion decreased on the first day, probably due to metabolism of remaining dietary protein. From the second day of starvation to the end of the starvation period, the urinary nitrogen excretion increased progressively. The hepatic glycogen stores were depleted at the end of the first day. The blood glucose concentration remained constant throughout starvation period except for a 15% decrease on the first day. There was increased mobilization of lipid stores, starting on the first day, reflected by an increase in the blood free fatty acids, glycerol and ketone body concentrations. These metabolite concentrations began to increase on the third and fourth day which probably reflected depleted fat stores since no visible body fat was observed by the fourth day. The data indicate that the rat does not spare body protein during starvation, probably because it depletes its glycogen and fat stores rapidly and must then depend on body protein as the major fuel for energy metabolism.

Amino Acids↗
PubMed1977

On the mechanism of glucagon stimulation of hepatic gluconeogenesis.

The addition of L-alanine as substrate to a perfused rat liver preparation produced a five-fold increase in the rate of glucose production. This enhancement of the gluconeogenic flux seems to be a consequence of a rise in the steady-state levels of pyruvate and oxaloacetate subsequent to the rise in alanine concentration. Glucagon (2 X 10(-9) M) increased the gluconeogenic flux from alanine (10 mM) by 50 percent, even though the concentration of the substrate in the perfusion fluid was at saturation. This effect was accompanied by a rise in the intracellular concentration of alanine. However, the steady-state concentration of pyruvate and oxaloacetate were decreased, probably as a consequence of a more reduced state of the nicotinamide-nucleotide system. In vivo, the intraperitoneal administration of glucagon to starved rats was accompanied by a decrease in the hepatic alanine and pyruvate concentrations despite the striking effects raising the plasma glucose levels. These observations seem to indicate that the effect of the hormone increasing the hepatic glucose output must be mediated through some other mechanism(s) independent of the intracellular variations in the hepatic amino acids levels.

Alanine↗