Biomedical subjects

J Satrústegui

Publications and source records attributed to J Satrústegui.

At least 55 records · Page 3Linked to original sources

PubMed1987

Variations due to hyperoxia and ageing in the activities of glutathione S-transferase and NADPH-cytochrome c reductase.

We have studied the influence of hyperoxia and ageing on the activities of NADPH-cytochrome c reductase and glutathione S-transferase in different rat organs. Lung glutathione S-transferase activity increases markedly in 5-day-old pups exposed to hyperoxia, as observed for the O2- scavenging enzyme, superoxide dismutase. The levels of NADPH-cytochrome c reductase increase as well but after a 3-day lag period. In the liver, there is a pronounced decrease of both activities in 24-month-old rats, but at 12 months the activity of glutathione S-transferase increases whereas that of NADPH cytochrome c reductase activity decreases with respect to 3 months. The pattern of variations with age of NADPH cytochrome c reductase is similar in liver and brain. However the behaviour of brain glutathione S-transferase parallels that of the liver enzyme only up to 12 months. Thereafter the brain activity is maintained at a high level. These observations open the possibility that the high glutathione S-transferase levels in the old rat brain might be involved in protection towards oxidative alterations during ageing.

Aging↗

PubMed1986

The influence of age on the calcium-efflux pathway and matrix calcium buffering power in brain mitochondria.

The variations with age of the ruthenium red-insensitive calcium efflux rate have been studied in rat brain mitochondria. Both H+- and Na+-dependent effluxes are decreased with age when expressed as a function of calcium taken up in mitochondria incubated in the presence of 0.8 mM inorganic phosphate (Pi) and 0.2 mM ADP. However, the age-dependent differences in calcium efflux rates disappear when mitochondria are incubated in the absence of ADP and Pi. It is suggested that the decrease in efflux rate observed with age corresponds to an increased calcium buffering power of the mitochondrial matrix due to an increase in mitochondrial Pi. The causes of the increased Pi accumulation in old-rat-brain mitochondria are yet unknown but possibly not due to differences in the Pi efflux. The results suggest that the age-dependent lowering of the free calcium concentration in the brain mitochondrial matrix together with the reduced activity of the calcium uniporter (Vitórica, J. and Satrústegui, J. (1986) Brain Research 378, 36-48) could lead to an impaired activation of mitochondrial dehydrogenases after a rise in cytosolic calcium.

Aging↗

PubMed1986

Involvement of mitochondria in the age-dependent decrease in calcium uptake of rat brain synaptosomes.

Calcium uptake in rat brain synaptosomes decreases during ageing. The possible involvement of mitochondria in altered calcium homeostasis has been investigated. Mitochondria isolated from old rat brain showed decreased calcium uptake rates. Since neither the mitochondrial membrane potential nor the delta pCa decreases with age, it was concluded that variations in the driving force for calcium uptake were not the cause for impaired calcium transport in mitochondria from aged rat brain. The steady state calcium distribution in isolated aged rat brain mitochondria was achieved at higher extramitochondrial calcium concentrations than that of adults. Studying the effects of the selective release of calcium from the mitochondrial pool by the addition of an uncoupler to 45Ca loaded synaptosomes incubated in high-potassium media, it was found that the intrasynaptic mitochondrial pool and the intra/extramitochondrial 45Ca distribution also decreased considerably in 24-month-old rats. Steady state fluorescence anisotropy (rs) of diphenylhexatriene-labelled mitoplasts from 'free' brain mitochondria increased with ageing. However, since no changes in rs from synaptosomal mitochondria were found in 24-month-old rats, it is suggested that alterations in lipid dynamics are not involved in the impaired calcium uptake observed in brain mitochondria from aged rats. The implications of these findings in the calcium homeostasis of brain endings are discussed.

Aging↗

PubMed1986

Increased basal gluconeogenesis in the aged rat.

Post-absorptive gluconeogenesis from lactate measured in vivo increases 3-fold in 24-month-old rats compared to 3-month-old animals. Fractional lactate turnover rates showed no significant differences between the two groups of animals. Lower plasma glucose concentrations and insulin-glucagon ratios may explain the increase in gluconeogenesis observed in aged rats.

Aging↗

PubMed1985

The role of ADP in the modulation of the calcium-efflux pathway in rat brain mitochondria.

The role of ADP in the regulation of Ca2+ efflux in rat brain mitochondria was investigated. ADP was shown to inhibit Ruthenium-Red-insensitive H+- and Na+-dependent Ca2+-efflux rates if Pi was present, but had no effect in the absence of Pi. The primary effect of ADP is an inhibition of Pi efflux, and therefore it allows the formation of a matrix Ca2+-Pi complex at concentrations above 0.2 mM-Pi and 25 nmol of Ca2+/mg of protein, which maintains a constant free matrix Ca2+ concentration. ADP inhibition of Pi and Ca2+ efflux is nucleotide-specific, since in the presence of oligomycin and an inhibitor of adenylate kinase ATP does not substitute for ADP, is dependent on the amount of ADP present, and requires ADP concentrations in excess of the concentrations of translocase binding sites. Brain mitochondria incubated with 0.2 mM-Pi and ADP showed Ca2+-efflux rates dependent on Ca2+ loads at Ca2+ concentrations below those required for the formation of a Pi-Ca2+ complex, and behaved as perfect cytosolic buffers exclusively at high Ca2+ loads. The possible role of brain mitochondrial Ca2+ in the regulation of the tricarboxylic acid-cycle enzymes and in buffering cytosolic Ca2+ is discussed.

Adenosine Diphosphate↗

PubMed1985

Impairment of glutamate uptake and absence of alterations in the energy-transducing ability of old rat brain mitochondria.

The proton electrochemical gradient has been measured in old brain mitochondria isolated from 2- or 24-month-old rats with the use of different respiratory substrates. With succinate as substrate, neither the respiratory rate, membrane potential or delta pH varied with age, indicating that the dielectric strength of the mitochondrial membrane was unaltered in old animals. The ohmic behavior of the membrane was tested in experiments in which the respiratory rate was partially inhibited by malonate, and was found to be unchanged with age. When glutamate plus malate were used as substrates, the respiratory rate was substantially reduced, and a drastic decrease in glutamate uptake was observed in old rat brain mitochondria.

Aging↗

PubMed1985

Age-dependent modifications of rat heart succinate dehydrogenase.

The activity of rat heart succinate dehydrogenase (SDH) increases 1.7-fold in old animals (Vitorica, J., Cano, J., Satrústegui, J. and Machado, A., Mech. Ageing Dev., 16 (1981) 105-116). This increase is due to an increase in enzyme protein itself because: (a) the activation state of the enzyme does not vary with age; and (b) the increase of activity is paralleled by an increase in immunoprecipitable SDH in old rat heart mitochondria. SDH from old rat heart mitochondria differs in a number of ways from that of young animals: (a) The km value for succinate increases with age. (b) The thermostability decreases, and the activation energy in the 20-40 degrees C interval is higher in old animals. (c) The breaking points of the Arrhenius plots of SDH are shifted to higher values. (d) Reactivity towards N-ethylmaleimide in succinate protected mitochondria decreases with age.

Aging↗

PubMed1985

Regulation by ammonium of glutamate dehydrogenase (NADP+) from Saccharomyces cerevisiae.

The activity of glutamate dehydrogenase (NADP+) (EC 1.4.1.4; NADP-GDH) of Saccharomyces cerevisiae is decreased under conditions in which intracellular ammonia concentrations increases. A high internal ammonia concentration can be obtained (a) by increasing the ammonium sulphate concentration in the culture medium, and (b) by growing the yeast either in acetate + ammonia media, where the pH of the medium rises during growth, or in heavily buffered glucose + ammonia media at pH 7.5. Under these conditions cellular oxoglutarate concentrations do not vary and changes in NADP-GDH activity appear to provide a constant rate of oxoglutarate utilization. The following results suggest that the decrease in NADP-GDH activity in ammonia-accumulating yeast cells is brought about by repression of synthesis: (i) after a shift to high ammonium sulphate concentrations, the number of units of activity per cell decreased as the inverse of cell doubling; and (ii) the rate of degradation of labelled NADP-GDH was essentially the same in ammonia-accumulating yeast cells and in controls, whereas the synthesis constant was much lower in the ammonia-accumulating cells than in the controls.

Acetates↗

PubMed1984

Interaction with protein SH groups could be involved in adriamycin cardiotoxicity.

The use of the antineoplastic agent adriamycin is limited by its cardiotoxicity. The mechanism of cardiotoxicity has been investigated through the study of adriamycin effects on a number of heart enzymes. Adriamycin inhibited the activity of NADP-isocitrate dehydrogenase, malic enzyme, and 6-phosphogluconate dehydrogenase, three enzymes that have in common the presence of reactive SH groups involved in activity. Adriamycin action was prevented by the presence of proteins or dithioerythrol and mimicked by dithiobis dinitrobenzoate. It is suggested that adriamycin effects are due to interaction with enzyme SH groups by a product of adriamycin metabolism.

Animals↗

PubMed1984

Age-dependent variations in peroxide-utilizing enzymes from rat brain mitochondria and cytoplasm.

The location of peroxide-utilizing enzymes has been studied in rat brain. Glutathione peroxidase and glutathione reductase distributions indicate that both enzymes are located in the cytoplasm and in the matrix space of "synaptosomal" and "free" mitochondria. On the other hand, catalase distribution parallels that of NADH-cytochrome c reductase (rotenone-insensitive), and appears to be associated with the outer membrane of brain mitochondria. Whereas no gross age-dependent changes in various marker enzymes were found, a gradual but significant increase in glutathione peroxidase from the soluble fraction of free mitochondria was detected. The consequences of such increase are discussed with regard to the reducing potential of the cell.

Aging↗

PubMed1984

Development of enzymes of energy metabolism in rat heart.

The behavior of several enzymes was studied during rat heart development (4 days before birth to adult stage). Hexokinase has its highest activity during the fetal period; it decreases at birth and remains with low activity in the adult. The alpha-glycerophosphate dehydrogenase and alpha-glycerophosphate oxidase profiles are similar up to the 15th day of development. From there onwards, both profiles diverge, the cytoplasmic activity increasing 3-fold, while the mitochondrial activity remains unchanged. The developmental profiles of the malate dehydrogenases are almost parallel. The development of citrate synthase and succinate dehydrogenase results in a 2- to 4-fold increase in their activities. However, ATPase increases dramatically (20-fold) over the same period. With respect to the enzymes of the adenine nucleotide metabolism, adenylate kinase is fully expressed throughout all ages examined, showing no variation during development. AMP deaminase and creatine kinase increase during development, the cytoplasmic creatine kinase reaching a high level at birth whereas the increases of the mitochondrial enzymes take place gradually during development.

AMP Deaminase↗

PubMed1983

Ammonia accumulation in acetate-growing yeast.

During growth on acetate, the pH of yeast cultures rises from 5.8 to around 7-8 in the stationary phase. This was found to result from acetic acid uptake and accompanying H+ loss. In addition, acetate-growing yeast were found to accumulate ammonia. The influence of pH on ammonia transport and accumulation was studied with the analogue [14C]methylamine with the following results. (a) Methylamine uptake kinetics from 0.1-50 mM were consistent with a single-component uptake system (NH+4 permease) at pH values more acidic than 6.5, and with a two-component system (NH+4 permease and NH3 diffusion) above pH 7.5. (b) Equilibrium accumulation of methylamine was found to increase with increasing pH. (c) Methylamine efflux from methylamine-loaded cells increased as the external pH decreased. It was concluded from measurements of the internal pH under various culture conditions that the accumulation of ammonia in acetate-growing alkaline cultures resulted from the sum of two processes: (1) an energy-driven NH+4 transport; and (2) NH3 diffusion dependent on the delta pH.

Acetates↗

PubMed1983

Citrate synthase of Tetrahymena pyriformis: evolutionary and regulatory aspects.

Citrate synthase (EC 4.1.3.7) from Tetrahymena pyriformis has been purified 185-fold. The molecular weight of the native enzyme was determined to be 120,000. The enzyme is labile at low ionic strength, but can be stabilized by KCl and glycerol. It is activated by KCl at low (below 60 mM) or high concentrations, and inhibited by divalent cations (Mn2+, Mg2+, Ca2+). The Michaelis constants are 0.1 mM for oxalacetate and 0.01 mM for acetyl-CoA. The kinetics with oxalacetate exhibit negative cooperativity, with a nH = 0.66. Among the metabolites tested, only ATP and GTP can inhibit the enzyme but Mg2+ relieves the ATP inhibition. Incubation with sulfhydryl reagents (DTNB) in the absence of its substrates results in a rapid inactivation of the enzyme. It is concluded that Tetrahymena citrate synthase is closer to the enzyme from Gram-positive bacteria than to those of eucaryotes.

Adenosine Triphosphate↗

PubMed1983

Development of NADPH-consuming pathways in heart, brain and liver of the rat.

The behavior of the principal NADPH-consuming pathways, fatty acid synthesis (-ATP-citrate lyase), transhydrogenase, glutamate dehydrogenase, glutathione reductase, NADPH-cytochrome c reductase and mitochondrial and cytoplasmic thioredoxin reductases were studied during the development of the heart, brain and liver of the rat. The liver is the tissue with highest NADPH-consuming activities. Transhydrogenase activity is highest in the heart; it undergoes two steep increases of activity, one at birth and another between 15 and 25 days. Liver cytoplasmic glutathione reductase activity increases mainly at birth. The activities in heart and brain are lower and the brain enzyme activity slightly increases throughout development. Liver NADPH-cytochrome c reductase activity greatly increases at weaning. The heart activity is the lowest, no changes taking place in the developmental period. Heart mitochondrial and brain cytoplasmic thioredoxin reductases undergo increases in activity at birth. The ratios of NADPH-producing/NADPH-consuming activities are constant in all tissues studied, and oscillate between congruent to 4 in fetuses to congruent to 2 in adults, the only exception being the adult heart ratio (congruent to 23). This otherwise constancy confirms that both processes are closely correlated.

ATP Citrate (pro-S)-Lyase↗

PubMed1982

[Study of the inhibition produced by adriamycin against specific enzymes in the rat heart].

Adriamycin is used for the treatment of leukemia and other neoplastic processes. Unfortunately the drug has a toxic effect on some tissues. Cardiotoxicity in particular limits the use of the drug. Several hypotheses have been given to explain adriamycin heart toxicity. The authors have studied the effect of the drug on the enzymes isocitrate dehydrogenase-NADP, malic enzyme, 6-P-gluconate dehydrogenase, malic dehydrogenase-NAD+, hexokinase, and phosphofructokinase. They observed that adriamycin inhibits the NADP-dependent enzymes, and that the sulfhydryl group of the enzymes may be involved in the inhibitory action of adriamycin.

Animals↗

PubMed1981

Age-related quantitative changes in enzyme activities of rat brain.

The patterns of brain enzymes linked to energy metabolism have been determined in rats aged between 3 and 21 months and compared to those of the developing brain as an estimate of the senescent energy capacity of this organ. During aging, pyruvate kinase increases, pointing towards an enhancement of the glucose-dependence of this organ. However, NAD-isocitrate dehydrogenase declines, suggesting a reduction of Krebs cycle activity in the aged rat brain. An increase in cytoplasmic NAD-malate dehydrogenase found during aging could provide an alternative mechanism of NAD recovery.

Aging↗

PubMed1981

Comparison between developmental and senescent changes in enzyme activities linked to energy metabolism in rat heart.

The developmental and senescent patterns of a number of heart enzyme activities linked to energy metabolism have been studied in rats aged between 4 days and 21 months. A morphometric study of mitochondrial volume fractions and numbers has been also carried out. Developmental changes result in a rise of most mitochondrial enzymes (NADP+-isocitrate dehydrogenase, malic enzyme, succinate dehydrogenase, citrate synthase) and mitochondrial volume fractions. Exceptions are NAD+-isocitrate dehydrogenase, which declines from 4 days onwards, and NAD+-malate dehydrogenase, which declines and then rises over the same period. Senescent changes follow two different trends. While pyruvate kinase and those mitochondrial enzymes lying between citrate formation and isocitrate oxidation (citrate synthase, NADP+-and NAD+-isocitrate dehydrogenases) decline to some degree, mitochondrial succinate dehydrogenase and NAD+-malate dehydrogenase activities increase over the same period. This could point towards a partial impairment of Krebs cycle function, and a reduced energy-producing capacity in the aged rat heart.

Aging↗

PubMed1981

Metabolic implications of ageing: changes in activities of key lipogenic and gluconeogenic enzymes in the aged rat liver.

The variations in enzyme activities involved in the main pathways of liver energetic metabolism--glycolysis, Krebs cycle, gluconeogenesis and lipogenesis--have been studied in rats ranging between the age of 4 days and 21 months. The major changes observed are the following: (1) enzymes involved in glycolysis (pyruvate kinase) and lipogenesis (NADP-malic enzyme, ATP-citrate lyase) decrease in activity during ageing, and (2) gluconeogenic enzymes (phosphoenolpyruvate carboxykinase, glucose-6-phosphatase) are maintained or slightly increased over the same period. The results suggest that an increase in the capacity for gluconeogenesis with respect to that for lipogenesis takes place in the aged rat liver.

ATP Citrate (pro-S)-Lyase↗