Ammonia effects on pyruvate/lactate production in astrocytes--interaction with glutamate

Ammonia exerts a multitude of metabolic and non-metabolic effects on brain tissue. In the present communication we have investigated its effect on lactate production rates, pyruvate production rates and pyruvate/lactate ratios in mouse cerebrocortical astrocytes and neurons in primary cultures. No effects were found in neurons. All three parameters were affected by ammonia in astrocytes, but less potently and to a smaller degree in cells that had been treated with dibutyryl cyclic AMP (morphologically differentiated cells) than in untreated cells (morphologically undifferentiated cells). In the differentiated cells ammonia had virtually no effect up to a concentration of 1.0 mM, but at 3.0 mM it increased lactate production and decreased pyruvate/lactate ratio significantly. In the undifferentiated cells ammonia greatly increased lactate accumulation (by 80% at 3.0 mM) and it inhibited pyruvate accumulation (by 40% at 3.0 mM). It thereby reduced the pyruvate/lactate ratio progressively within the entire range 0.1-3.0 mM ammonia. In support of the hypothesis that the ammonia-induced reduction of pyruvate/lactate ratio is secondary to depletion of cellular glutamate by formation of glutamine (and glutathione) and a resulting interruption of the malate-aspartate shuttle (MAS), the addition of glutamate to the incubation medium significantly diminished the ammonia-induced reduction of pyruvate/lactate ratio, whereas it had no effect on the increased lactate production. It is discussed that MAS interruption may have additional consequences in astrocytes.     

The movement of pyruvate, lactate and lactate dehydrogenase into rabbit oviductal fluid.

Pyruvate, lactate and lactate dehydrogenase appeared linearly in 2 ml 0.9% NaCl recirculated through the rabbit oviduct for 4 h in vivo. In oviducts from rabbits injected 3 days previously with 100 i.u. hCG, the rate of appearance of all three constituents was considerably reduced. It is considered unlikely that the lactate dehydrogenase secreted brings about the interconversion of pyruvate and lactate in the oviduct lumen.     

Citrate, pyruvate, and lactate contaminants of commercial serum albumin

Commercial serum albumin was found to contain lactate, pyruvate, and especially citrate in addition to fatty acids. Glucose, aspartate, and alpha-ketoglutarate were also present but at lower concentrations. Charcoal treatment followed by prolonged dialysis was effective in removing most of these contaminants.     

Metabolism of pyruvate and citrate in lactobacilli

Lactobacillus acidophilus, L. bulgaricus, L. casei, L. delbrueckii , L. lactis and L. plantarum contained a pyruvate oxidase for the oxidation of pyruvate to acetyl phosphate and acetate. The presence of an acetate kinase converted the acetyl phosphate to acetate. L. casei and L. plantarum produced lactate and acetoin, in addition to acetate, under the conditions used while L. casei also produced diacetyl. L. casei and L. plantarum were the only species to utilize citrate. L. helveticus and L. helveticus subsp. jugurti did not utilize pyruvate under the conditions used.     

Fermentation of trans-aconitate via citrate,oxaloacetate, and pyruvate by Acidaminococcus fermentans

Growing cells of Acidaminococcus fermentans (DSM 20731 and ATCC 25085) fermented trans-aconitate via citrate, oxaloacetate, and pyruvate to approximately 2 CO₂, 1.8 acetate, 0.1 butyrate and 0.9 H₂. The carbon and electron recoveries were close to 100%. On citrate no growth was observed and washed cells were unable to ferment this tricarboxylate. In cell-free extracts, however, citrate as well as trans-aconitate were readily fermented to CO₂ and acetate. Under these conditions, also cis-aconitate, oxaloacetate, and pyruvate were formed, whereas butyrate and intermediates of glutamate fermentation, 2-oxoglutatrate and glutaconate, could not be detected. Citrate Si-lyase, a Mg²⁺-dependent oxaloacetate decarboxylase, and pyruvate synthase were present in quantities that corresponded to the growth rate of the organism. Received: 3 May 1996 / Accepted: 12 August     

Studies on the effects of lactate transport inhibition, pyruvate, glucose and glutamine on amino acid, lactate and glucose release from the ischemic rat cerebral cortex

A rat four vessel occlusion model was utilized to examine the effects of ischemia/reperfusion on cortical window superfusate levels of amino acids, glucose, and lactate. Superfusate aspartate, glutamate, phosphoethanolamine, taurine, and GABA were significantly elevated by cerebral ischemia, then declined during reperfusion. Other amino acids were affected to a lesser degree. Superfusate lactate rose slightly during the initial ischemic period, declined during continued cerebral ischemia and then was greatly elevated during reperfusion. Superfusate glucose levels declined to near zero levels during ischemia and then rebounded beyond basal levels during the reperfusion period. Inhibition of neuronal lactate uptake with alpha-cyano-4-hydroxycinnamate dramatically elevated superfusate lactate levels, enhanced the ischemia/reperfusion evoked release of aspartate but reduced glutamine levels. Topical application of an alternative metabolic fuel, glutamine, had a dose dependent effect. Glutamine (1 mM) elevated basal superfusate glucose levels, diminished the decline in glucose during ischemia, and accelerated its recovery during reperfusion. Lactate levels were elevated during ischemia and reperfusion. These effects were not evident at 5 mM glutamine. At both concentrations, glutamine significantly elevated the superfusate levels of glutamate. Topical application of sodium pyruvate (20 mM) significantly attenuated the decline in superfusate glucose during ischemia and enhanced the levels of both glucose and lactate during reperfusion. However, it had little effect on the ischemia-evoked accumulation of amino acids. Topical application of glucose (450 mg/dL) significantly elevated basal superfusate levels of lactate, which continued to be elevated during both ischemia and reperfusion. The ischemia-evoked accumulations of aspartate, glutamate, taurine and GABA were all significantly depressed by glucose, while phosphoethanolamine levels were elevated. These results support the role of lactate in neuronal metabolism during ischemia/reperfusion. Both glucose and glutamine were also used as energy substrates. In contrast, sodium pyruvate does not appear to be as effectively utilized by the ischemic/reperfused rat brain since it did not reduce ischemia-evoked amino acid efflux.     

Dynamics of oxyhemoglobin, lactate, and pyruvate in plasma in connection with the woman menstrual cycle

Levels of lactate, piruvate, oxy- and carbohaemoglobin were shown to change regularly during women's menstrual cycle reflecting changes in metabolism in respect to forming and destroying of the ovum.     

Transport of pyruvate and lactate in yeast mitochondria.

Evidence for the existence of mediated transport of pyruvate and lactate in isolated mitochondria of Saccharomyces cerevisiae is presented. 1. The mitochondrial oxidation of pyruvate is specifically inhibited by the monocarboxylic oxoacids alpha-ketoisocaproate and by alpha-cyano-3-hydroxycinnamate, while pyruvate and malate dehydrogenases activities are not inhibited. 2. The stimulation of the mitochondrial oxidations of succinate, alpha-ketoglutarate and citrate by pyruvate are also inhibited by alpha-cyano-3-hydroxycinnamate. 3. The [14C]pyruvate uptake by yeast mitochondria follows saturation kinetics and is completely inhibited by alpha-cyano-3-hydroxycinnamate. 4. Large amplitude passive swellings of mitochondria of the wild type and of cytoplasmic rho- and rho-n mutants are induced by isoosmotic ammonium pyruvate and lactate. These pH-dependent swellings are inhibited by alpha-cyano-3-hydroxycinnamate suggesting that the carrier system is not coded by mitochondrial DNA.     

4-dimethylamino-3',4'-dimethoxychalcone downregulates iNOS expression and exerts anti-inflammatory effects

Reactive oxygen and nitrogen species contribute to the pathophysiology of inflammatory conditions. We have studied the effects of a novel superoxide scavenger, 4-dimethylamino-3', 4'-dimethoxychalcone (CH11) in macrophages and in vivo. CH11 has been shown to inhibit the chemiluminescence induced by zymosan in mouse peritoneal macrophages and the cytotoxic effects of superoxide. In the same cells, the modulation by superoxide of nitric oxide (NO) production in response to zymosan was investigated. CH11 was more effective than the membrane-permeable scavenger Tiron for inhibition of inducible nitric oxide synthase (iNOS) protein expression and nitrite production. We have shown that CH11 inhibited chemiluminescence in vivo, as well as cell migration, and eicosanoid and tumor necrosis factor-alpha (TNF-alpha) levels in the mouse air pouch injected with zymosan. This chalcone derivative also exerted anti-inflammatory effects in the carrageenan paw oedema.     

The intracellular location of pyruvate carboxylase, citrate synthase and 3-hydroxyacyl-CoA dehydrogenase in lactating rat mammary gland.

The intracellular location of pyruvate carboxylase (EC 6.4.1.1), citrate synthase (EC 4.1.3.7) and 3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) in rat mammary gland was investigated by using a fractional-extraction technique. The results indicate a mitochondrial location for all three enzymes.     

LDH isoenzyme pattern, lactate/pyruvate, and ultrastructure of fibrinogenic and epithelial cell lines exposed to 3% oxygen

Human diploid fibroblast WI38 and canine epithelial MDCK cells were incubated in an environment flushed continuously for 96 h with 3% and 20% oxygen. Lactate and pyruvate formation and LDH isoenzyme patterns of cells in logarithmic and stationary phases showed characteristic differences between both cell lines. Lactate formation by WI38 cells at 3% oxygen was considerable within the first 18 h. The MDCK cells responded with an even greater lactate formation only after 18 h.The dynamics of LDH isoenzyme changes measured as H/M subunits revealed for WI38 cells a sharp decline after 40 h at 3% oxygen in both logarithmic and confluent stages. MDCK cells show a sharp decline in H/M during the first 40 h (confluent phase) or 20 h (log phase). No changes in either cell line, regardless of the oxygen environment, were seen in the ultrastructure of cells, as seen by transmission electron microscopy. It is suggested that the fibroblast, being better equipped for anaerobic metabolism, quickly produces more lactate without changing the LDH isoenzyme pattern at 3% oxygen. Epithelial cells react first with a modification in the LDH pattern and then a continuous increase in the formation of lactate to very high values.