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PubMed1995

Structure of simusan, a new acidic exopolysaccharide from Arthrobacter sp.

Simusan, a major exopolysaccharide produced by an ethanol-utilizing Arthrobacter sp. strain CE-17, contains D-glucose, D-mannose, D-galactose, L-rhamnose, D-glucuronic acid, and pyruvic acid in the ratios approximately 3:2:1:1:1:1 as well as O-acyl groups (presumably (presumably residues of acetic and palmitic acid). On the basis of chemical modifications of the polysaccharide, solvolysis with anhydrous hydrogen fluoride resulting in a penta- and an octa-saccharide fragment, Smith degradation, and 1H and 13C NMR analysis, the following structure of the repeating unit was established: [formula: see text] It is suggested that at least one of the glucose residues and the galactose residue are O-acetylated.

Arthrobacter↗
PubMed1986

Effect of the mycotoxin citrinin on some hormones and on enzymes and substrates of the Embden-Meyerhof pathway in mice.

The effects of six weeks treatment with multiple i.p. doses of citrinin (15, 25 and 35 mg/kg) on some hormonal activities and on enzymes and substrates of the Embden--Meyerhof pathway in mice were studied. Adenosine triphosphatase, hexokinase, lactate dehydrogenase, lactic acid and pyruvic acid decreased in blood, liver, kidney and brain of citrinin-treated mice. Glucose levels in blood, kidney and brain increased and glycogen content of liver decreased. Cortisol, triiodothyronine and thyroxine levels of serum increased, but insulin levels decreased.

Anaerobiosis↗
PubMed1991

Effect of human, recombinant interleukin 2 on Punta Toro virus infections in C57BL/6 mice.

The effect of human recombinant interleukin-2 (rIL-2) on Punta Toro virus (PTV) infection was investigated in C57BL/6 mice. Immunologic and viral parameters were assessed after mice were treated i.p. with rIL-2 for 5 days. Treatment of mice with 25000 and 12500 units/mouse of rIL-2 resulted in significant inhibition of the disease as indicated by increases in survival of mice as well as decreases in liver and serum virus titers. Serum glutamic oxalic acid and pyruvic acid transaminase levels were also lowered indicating reduced liver damage. Murine IL-2 production returned to normal or above-normal levels in rIL-2 treated mice. Natural killer cell activity was also moderately stimulated by rIL-2 treatment. Significant amounts of interferon were not detected in the sera of treated mice. Weight gain and survival rates were similar for both toxicity and normal controls indicating that rIL-2 treatments had no toxic effect.

Animals↗
PubMed1989

Modulation of prostaglandin E2 catabolism and action by fuel substrates in rat hepatocytes.

The hepatic level of prostaglandins will reflect the balance between synthesis of prostaglandins and their rapid catabolism via beta-oxidation by hepatocytes. In the present study we examined the effect of physiological fuel substrates on the breakdown and action of prostaglandin E2 (PGE2) in isolated rat hepatocytes. Palmitic acid (0.32 mM), a long-chain fatty acid, inhibited the rate of PGE2 breakdown (10(-7) M) by approx. 80%. As the palmitic acid concentration was increased from 0 to 0.8 mM, the percentage of PGE2 remaining in the incubation 5 min following prostaglandin addition was raised from approx. 10% to over 98%. Octanoic acid (0.8 mM) also inhibited PGE2 catabolism, while butyric acid (0.8 mM) and pyruvic acid (2.5 mM) were without effect. The inhibition of glucagon-stimulated glycogenolysis by PGE2 was increased in the presence of 0.6 mM palmitic acid, consistent with decreased PGE2 catabolism. These studies demonstrate that changes within the range of free fatty acid concentrations seen physiologically in vivo may dramatically alter PGE2 catabolism and, therefore, the effect of PGE2 to modulate hormonal action in the liver.

Animals↗
PubMed1981

Biochemical studies in infective amosite pneumoconiosis.

The effects of Candida albicans, of amosite dust, and of the 2 agents combined, on some biochemical parameters of the lung in male guinea pigs were determined. Adult guinea pigs were intratracheally injected with 50 mg of amosite dust and 0.3 mg of viable suspension of mycelia of Candida albicans. The other groups received either dust or organism or none. Biochemical measurements done at 30, 60 and 90 days after infection showed that the superimposed state produced more rapid and drastic changes in pulmonary contents of collagen, mucopolysaccharides and phospholipids and activities of lactic dehydrogenase and succinate dehydrogenase. The levels of lactic acid and pyruvic acid were also more profoundly changed after amosite + Candida. These results indicate that Candida albicans which is commonly found in upper respiratory tract could aggrevate the lesions caused by exposure to asbestos.

Animals↗
PubMed1996

Indirect ultraviolet detection of biologically relevant organic acids by capillary electrophoresis.

Indirect UV detection of fourteen short-chain organic acids (e.g., oxalic acid, citric acid, malonic acid, tartaric acid, methylmalonic acid, alpha-ketoglutaric acid, succinic acid, ethylmalonic acid, methylsuccinic acid, glutaric acid, apidic acid, methylglutaric acid, lactic acid and pyruvic acid) by capillary electrophoresis is described. The method used phthalate as the UV-absorbing additive in carbonate buffer and the non-absorbing analytes were detected indirectly at 230 nm. The influences of buffer pH, ionic strength, concentration of phthalate and organic modifier on indirect signal response and migration behavior of the organic acids were investigated. Comparisons of reproducibility on migration time, limit of detection and separation efficiency among three types of capillary (e.g., polyacrylamide-coated, myristyltrimethylammonium bromide-coated and uncoated capillaries) were conducted. The method developed was applied to detect succinic acid, methylmalonic acid, citric acid, glutaric acid and lactic acid in human body fluids, and preliminary results were provided.

Buffers↗
PubMed2006

Structural analysis of an extracellular polysaccharide produced by a benzene tolerant bacterium, Rhodococcus sp. 33.

Rhodococcus sp. 33 can tolerate and efficiently degrade various concentrations of benzene, one of the most toxic and prevailing environmental pollutants. This strain produces a large quantity of extracellular polysaccharide (33 EPS), which plays an important role in the benzene tolerance in Rhodococcus sp. 33, especially by helping the cells to survive an initial challenge with benzene. This EPS has been reported to be composed of D-galactose, D-glucose, D-mannose, D-glucuronic acid, and pyruvic acid at a molar ratio of 1:1:1:1:1. To understand the protective effect of 33 EPS, we determined its chemical structure by using 1H and 13C NMR spectroscopy including 2D DQF-COSY, TOCSY, HMQC, HMBC, and NOESY experiments. The polysaccharide was shown to consist of tetrasaccharide repeating units with the following structure: [structure: see text].

Benzene↗
PubMed2005

Functional characteristics of H+ -dependent nicotinate transport in primary cultures of astrocytes from rat cerebral cortex.

In the present study, we report the characteristics of H(+)-coupled nicotinate transport in primary cultures of astrocytes from rat cerebral cortex. The [(3)H]nicotinate transport in rat astrocytes increased up to a pH 5.5. The nicotinic acid uptake at pH 6.0 was both energy-dependent and saturable with a Michaelis constant (K(t)) of 2.8+/-0.4 mM and the maximal uptake rate (V(max)) of 31+/-3.2 nmol/mg protein/10 min. This process was reduced by a protonophore, carbonylcyanide p-trifluoromethoxyphenylhydrazone, and a typical monocarboxylate transporter (MCT) inhibitor, alpha-cyano-4-hydroxycinnamic acid, suggesting that nicotinate uptake by rat astrocytes is mediated by H(+)-coupled monocarboxylate transport system. [(3)H]Nicotinate transport in rat astrocytes was significantly inhibited by various monocarboxylic acids such as l-lactic acid and pyruvic acid with a relatively low affinity (K(i)>10 mM). On the other hand, the uptake process of l-lactic acid was also saturable with a high-affinity component (K(t)=0.27 mM) and a low-affinity component (K(t)=35.9 mM). Reverse transcription-PCR and Western blot analyses revealed that three MCT subtypes, MCT1/Slc16a1, MCT2/Slc16a7, and MCT4/Slc16a3, were expressed in these cells. Because l-lactate reduced to 67% of the nicotinate uptake even at 10mM, it is unlikely that nicotinate uptake in rat astrocytes is mediated by MCT1 and/or MCT2. These results provide biochemical evidence of a H(+)-coupled and saturable transport system, presumed to be a low-affinity monocarboxylate transporter MCT4 or other unknown H(+)-coupled monocarboxylate transport system, for nicotinate in rat cerebrocortical astrocytes.

Animals↗
PubMed1976

High-speed liquid chromatographic determination of phenylpyruvic acid.

A high-speed liquid chromatographic method has been developed for the determination of urinary phenylpyruvic acid. This acid is converted by treatment with naphthalene-2,3-diamine into 3-benzyl-2-hydroxybenzoquinoxaline, which is extracted into carbon tetrachloride for separation. 2-Mercaptoethanol is a useful stabilizer, and 2-chlorothioxanthone is a suitable internal standard. The method is specific, and the results are not affected by the presence of such 2-oxo-acids as pyruvic acid, 2-oxobutyric acid, 2-oxoglutaric acid, and 4-hydroxphenylpyruvic acid.

Chemical Phenomena↗
PubMed1992

Elucidation of the topological parameters of N-acetylneuraminic acid and some analogues involved in their interaction with the N-acetylneuraminate lyase from Clostridium perfringens.

A series of neuraminic acid derivatives modified in the side chain or at C-3, C-4 or C-5 were tested as substrates of inhibitors of N-acetylneuraminate lyase (EC 4.1.3.3) from Clostridium perfringens. The results, together with Km and Ki values reported previously, indicate that the region most important for the binding of sialic acids is an equatorial zone reaching from C-8 via the ring oxygen atom to C-4 of the sugar molecule, whereas the substituents at C-9 and C-5 may be varied to a higher extent without significantly disturbing enzyme action. It is shown that stereo-electronic factors are responsible for the immediate heterolytic fragmentation of the cyclic sialic acid into pyruvic acid and 2-acetamidomannose or a related C-6 sugar.

Clostridium perfringens↗
PubMed2004

Bone-constructing cells from ethmoid bone may have multilineage differentiation potential: preliminary report.

OBJECTIVE: In order to investigate multilineage differentiation in human cultured cells from ethmoid bone, we conducted a morphological study to examine adipogenic and chondrogenic differentiation. MATERIAL AND METHODS: After reaching confluence, cells underwent terminal adipogenic differentiation by treatment with 100 microM indomethacin, 0.5 mM 1-methyl-3-isobutylxanthine, 1 microM dexamethasone (DEX), 10 microg/ml insulin and 0.3% dimethylsulfoxide in a medium supplemented with 10% fetal bovine serum. Chondrogenic differentiation was attempted by centrifuging a pelleted micromass using transforming growth factor-beta3 (TGF-beta3), DEX, ascorbic acid (AA), pyruvate acid, proline, glucose and (ITS)-plus. RESULTS: The cultured cells displayed adipocyte but not chondrogenic lineage under these conditions. Considering the possibility that some differentiation potential may be lost with in vitro culture but maintained using another chondrogenic differentiation medium containing TGF-beta1, it is possible that cultured cells may have multilineage potential, including chondrogenic differentiation ability. CONCLUSIONS: These morphological abilities of human cultured cells may indicate the possibility of the existence of mesenchymal stem cells in sinus bone. If mesenchymal stem cells exist in ethmoid bone, they may play an important role in future research on the regulation mechanisms of human bone tissue.

Adipocytes↗
PubMed1978

[Effect of food deprivation on various parameters of rumen juice and blood of sheep].

In order to find out how short-time denutrition changes the concentration of some substances in the rumen fluid and the blood, tests with full-grown sheep were carried out. Fodder was withheld from sheep with inserted Jarrett fistulae for 48 hours after normal feeding. After 48 hours the animals were given concentrated fodder, after 52 hours exclusively hay. From the 72nd hour onwards the animals were provided with fodder as usual. Samples of the rumen fluid and blood samples were taken at the beginning of the test, after the last normal feeding and then in the 24th, 32nd, 48th, 52nd, 56th, 72nd and 96th hour. We could find out that, during the 48-hour denutrition, the pH-value of the rumen fluid turned alkaline and the concentrations of ammonia, volatile fatty acids and lactic acid decreased. The protein metabolism underwent a rapid change in the organism. The protein content of the blood plasma decreased, above all the albumin content, as well as the concentration of glycoproteins and volatile amino acids. Among the various amino acids, the concentration of glycine increased highly, that of alanine and valine just slightly. The concentration of most amino acids decreased or--of some of them remained the same. Among the paramters that are characteristic of lipid and carbohydrate metabolism, the total content of lipids and cholesterin decreased, and so did the concentration of blood sugar, lactic acid and pyruvic acid in the blood plasma. The results indicate that short-time denutrition has a considerable influence on the rumen fermentation and the intermediary metabolism of ruminants. The quickly arising lack of energy of ruminants slows down the protein synthesis and increases the glyconeogenesis from amino acids. The tissue is supplied with energy by the mobilisation of lipids.

Amino Acids↗
PubMed1976

The mechanism of hypoglycemia due to semistarvation in the rat.

A quantity of diet providing 50% of the energy and protein requirement was fed to 4 week old rats. After 3 weeks, the amount of food was reduced to 25% and resulted in hypoglycemia in most of the rats within 4 to 7 days. The rats were divided into three groups: 1) normal controls; 2) malnourished, exhibiting blood sugar levels more than 40 mg/100 ml; 3) malnourished with blood sugar concentration less than 40 mg/100ml. The plasma concentrations of energy providing substrates: free fatty acids, glycerol, pyruvic acid, lactic acid and alanine were significantly lower in the hypoglycemic group than in the other two groups, while the first two groups displayed no significant differences. Plasma insulin concentrations were reduced in the undernourished groups and the lowest levels were found in the hypoglycemic rats. The plasma corticosterone concentration was significantly greater in group 2 but only slightly higher in group 3 than in the controls. Blood glucose concentration did not increase after intravenous glucagon in group 3, but the response was restored 4 hours after glucose administration. Blood glucose did not increase after alanine infusion in group 3 and increased only moderately in group 2. There was a normal increase after glycerol infusion in both malnourished groups. It was concluded that in semistarvation, fatal hypoglycemia is due to the depletion of fat stores accompanied by impaired gluconeogenesis from alanine.

Alanine↗
PubMed2006

Lactobacillus casei metabolic potential to utilize citrate as an energy source in ripening cheese: a bioinformatics approach.

AIMS: To identify potential pathways for citrate catabolism by Lactobacillus casei under conditions similar to ripening cheese. METHODS AND RESULTS: A putative citric acid cycle (PCAC) for Lact. casei was generated utilizing the genome sequence, and metabolic flux analyses. Although it was possible to construct a unique PCAC for Lact. casei, its full functionality was unknown. Therefore, the Lact. casei PCAC was evaluated utilizing end-product analyses of citric acid catabolism during growth in modified chemically defined media (mCDM), and Cheddar cheese extract (CCE). Results suggest that under energy source excess and limitation in mCDM this micro-organism produces mainly L-lactic acid and acetic acid, respectively. Both organic acids were produced in CCE. Additional end products include D-lactic acid, acetoin, formic acid, ethanol, and diacetyl. Production of succinic acid, malic acid, and butanendiol was not observed. CONCLUSIONS: Under conditions similar to those present in ripening cheese, citric acid is converted to acetic acid, L/D-lactic acid, acetoin, diacetyl, ethanol, and formic acid. The PCAC suggests that conversion of the citric acid-derived pyruvic acid into acetic acid, instead of lactic acid, may yield two ATPs per molecule of citric acid. Functionality of the PCAC reductive route was not observed. SIGNIFICANCE AND IMPACT OF THE STUDY: This research describes a unique PCAC for Lact. casei. Additionally, it describes the citric acid catabolism end product by this nonstarter lactic acid bacteria during growth, and under conditions similar to those present in ripening cheese. It provides insights on pathways preferably utilized to derive energy in the presence of limiting carbohydrates by this micro-organism.

Acetic Acid↗
PubMed1980

Improved medium for selective isolation and enumeration of Bifidobacterium.

Petuely's selective medium for Bifidobacterium was improved by addition of riboflavin, nucleic acid bases, pyruvic acid, and nalidixic acid. The modified medium, when examined under strictly anaerobic conditions for efficient isolation of Bifidobacterium from human fecal samples, exhibited selective and high viable counts that were close to those found on the usual nonselective medium.

Actinomycetaceae↗
PubMed1983

Linalyl Acetate Is Metabolized by Pseudomonas incognita with the Acetoxy Group Intact.

Metabolism of linalyl acetate by Pseudomonas incognita isolated by enrichment culture on the acyclic monoterpene alcohol linalool was studied. Biodegradation of linalyl acetate by this strain resulted in the formation of linalool, linalool-8-carboxylic acid, oleuropeic acid, and Delta-4-acetoxy-4-methyl hexenoic acid. Cells adapted to linalyl acetate metabolized linalyl acetate-8-aldehyde to linalool-8-carboxylic acid, linalyl acetate-8-carboxylic acid, Delta-4-acetoxy-4-methyl hexenoic acid, and geraniol-8-carboxylic acid. Resting cell suspensions previously grown with linalyl acetate oxidized linalyl acetate-8-aldehyde to linalyl acetate-8-carboxylic acid, Delta-4-acetoxy-4-methyl hexenoic acid, and pyruvic acid. The crude cell-free extract (10,000 g of supernatant), obtained from the sonicate of linalyl acetate-grown cells, was shown to contain enzyme systems responsible for the formation of linalyl acetate-8-carboxylic acid and linalool-8-carboxylic acid from linalyl acetate. The same supernatant contained NAD-linked alcohol and aldehyde dehydrogenases involved in the formation of linalyl acetate-8-aldehyde and linalyl acetate-8-carboxylic acid, respectively. On the basis of various metabolites isolated from the culture medium, resting cell experiments, growth and manometric studies carried out with the isolated metabolites as well as related synthetic analogs, and the preliminary enzymatic studies performed with the cell-free extract, a probable pathway for the microbial degradation of linalyl acetate with the acetoxy group intact is suggested.

Journal Article↗
PubMed2001

Bordetella pertussis autoregulates pertussis toxin production through the metabolism of cysteine.

Pertussis toxin (Ptx) expression and secretion in Bordetella pertussis are regulated by a two-component signal transduction system encoded by the bvg regulatory locus. However, it is not known whether the metabolic pathways and growth state of the bacterium influence synthesis and secretion of Ptx and other virulence factors. We have observed a reduction in the concentration of Ptx per optical density unit midway in fermentation. Studies were conducted to identify possible factors causing this reduction and to develop culture conditions that optimize Ptx expression. Medium reconstitution experiments demonstrated that spent medium and a fraction of this medium containing components with a molecular weight of <3,000 inhibited the production of Ptx. A complete flux analysis of the intermediate metabolism of B. pertussis revealed that the sulfur-containing amino acids methionine and cysteine and the organic acid pyruvate accumulated in the media. In fermentation, a large amount of internal sulfate (SO4(2-)) was observed in early stage growth, followed by a rapid decrease as the cells entered into logarithmic growth. This loss was later followed by the accumulation of large quantities of SO4(2-) into the media in late-stage fermentation. Release of SO4(2-) into the media by the cells signaled the decoupling of cell growth and Ptx production. Under conditions that limited cysteine, a fivefold increase in Ptx production was observed. Addition of barium chloride (BaCl2) to the culture further increased Ptx yield. Our results suggest that B. pertussis is capable of autoregulating the activity of the bvg regulon through its metabolism of cysteine. Reduction of the amount of cysteine in the media results in prolonged vir expression due to the absence of the negative inhibitor SO4(2-). Therefore, the combined presence and metabolism of cysteine may be an important mechanism in the pathogenesis of B. pertussis.

Adenylate Cyclase Toxin↗
PubMed2003

Chemolithoorganotrophic growth of Nitrosomonas europaea on fructose.

The nitrifying bacterium Nitrosomonas europaea can obtain all its carbon for growth from CO(2) and all its energy and reductant for growth from the oxidation of NH(3) and is considered an obligate chemolithoautotroph. Previous studies have shown that N. europaea can utilize limited amounts of certain organic compounds, including amino acids, pyruvate, and acetate, although no organic compound has been reported to support the growth of N. europaea. The recently completed genomic sequence of N. europaea revealed a potential permease for fructose. With this in mind, we tested if N. europaea could utilize fructose and other compounds as carbon sources to support growth. Cultures were incubated in the presence of fructose or other organic compounds in sealed bottles purged of CO(2). In these cultures, addition of either fructose or pyruvate as the sole carbon source resulted in a two- to threefold increase in optical density and protein content in 3 to 4 days. Studies with [(14)C]fructose showed that >90% of the carbon incorporated by the cells during growth was derived from fructose. Cultures containing mannose, glucose, glycerol, mannitol, citrate, or acetate showed little or no growth. N. europaea was not able to grow with fructose as an energy source, although the presence of fructose did provide an energy benefit to the cells. These results show that N. europaea can be grown in CO(2)-free medium by using fructose and pyruvate as carbon sources and may now be considered a facultative chemolithoorganotroph.

Fructose↗