Role of citrate, lactate, pyruvate, and fatty acids in albumin-induced bone resorption.
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Animals↗
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Structural studies of an acidic galactomannan from the reference strain for Serratia marcescens serogroup O4.
An acidic, partially acetylated galactomannan has been isolated from the lipopolysaccharide of the reference strain (C.D.C. 864-57) for Serratia marcescens serogroup O4. From the results of methylation analysis, Smith degradations, and n.m.r. spectroscopic studies of the O-deacetylated polymer, it was concluded that the repeating unit has the structure shown, in which the acetal-linked pyruvic acid has the R configuration. The polymer is believed to confer O specificity on the organism, but not to constitute the side chain of the lipopolysaccharide. (formula; see text).
Acetylation↗
Microbial synthesis of L-[15N]leucine L-[15N]isoleucine, and L-[3-13C]-and L-[3'-13C]isoleucines studied by nuclear magnetic resonance and gas chromatography-mass spectrometry.
The preparation of leucine and isoleucine labeled with 15N and of site-specific 13C-labeled isoleucines is described. This method is based on the induction of the biosynthetic pathways specific for branched chain amino acids in glutamic acid producing bacteria, and controlled provision of stable isotope labeled precursors. Corynebacterium glutamicum (ATCC 13032), a glutamic acid overproducer, was incubated in leucine production medium which consisted of a basal medium supplemented with [15N]ammonium sulfate, glucose, and sodium alpha-ketoisocaproate. production of L-[15N]leucine reached 138 mumol/ml at an isotopic efficiency of 90%. It was purified and checked by proton NMR and GC-MS. The electron impact (EI) spectrum showed 95 atom% enrichment. The cultivation of C. glutamicum in a similar medium containing alpha-ketobutyrate yielded L-[15N]isoleucine at a concentration of 120 mumol/ml. The GC-MS EI and chemical ionization (CI) spectra confirmed enrichment of 96 atom% 15N as that of the labeled precursors. The biosynthesis of L-[13C]isoleucine was carried out by induced cells which were transferred to a similar medium in which [2-13C]- or [3-13C]pyruvic acid replaced glucose. 13C NMR of the product isoleucine revealed single-site enrichment at C-3 or at C-3' respective to the precursor [13C]pyruvate; i.e., C-3 was labeled from [2-13C]pyruvate and C-3' from [3-13C]pyruvate. Mass spectrometric analysis confirmed that all molecules were labeled only in one carbon. This site-specific incorporation of [13C]pyruvate is contrasted with the labeling pattern obtained when producing cells were supplied with [2-13C]acetate, instead of pyruvate, when most label was incorporated into carbons 3 and 3' of the same isoleucine molecule.
Actinomycetales↗
Biochemical and molecular characterization of fumarase from plants: purification and characterization of the enzyme--cloning, sequencing, and expression of the gene.
A cDNA EST clone encoding the C-terminal portion of Arabidopsis thaliana fumarase was identified by homology analysis. A fragment of cDNA encoding the N-terminal region of fumarase was amplified from a cDNA library using PCR and cloned. Genomic DNA corresponding to the coding region of fumarase was amplified and cloned. Arabidopsis fumarase was expressed as a chimeric fusion protein and polyclonal antibodies were generated. Fumarase was purified to near-homogeneity (over 600-fold) from etiolated Pisum sativum mitochondria. The identification of fumarase was confirmed by a combination of immunoblot and N-terminal amino acid sequencing. Kinetic analysis of highly purified fumarase yielded a KM(malate) of 0.45 mM and a Vmax(malate) of 650 mumol of fumarate/min/ mg. The pea fumarase was inhibited by the alpha-keto acids pyruvate and alpha-ketoglutarate at low millimolar concentrations. Adenylates were highly inhibitory; the degree of this inhibition was reduced in the presence of Mg2+, suggesting that uncomplexed adenylates are the inhibitory species.
Amino Acid Sequence↗
Structure of extracellular polysaccharides of Escherichia coli strains 36M, 72M, and 29M isolated from coligranuloma of chick intestine. I. Polysaccharide from E. coli 36M.
An extracellular polysaccharide was isolated from culture broth of Escherichia coli 36M, and fractionated on a column of Sephadex G-150 into two fractions; the high molecular weight portion (85% of the total polysaccharide) contained pyruvic acid, and showed a positive immune reaction with anti-Ps-I-serum obtained from a rabbit. The low molecular weight portion (15% of the total polysaccharide) showed a negative immune reaction. The methylation, Smith's degradation, partial acid hydrolysis and methanolysis of the higher molecular weight polysaccharide revealed a repeating structure as follows: (see article).
Animals↗
Pyruvate dehydrogenase deficiency in a child responsive to thiamine treatment.
We report the clinical features in a 4-year-old child who was investigated for a suspected metabolic disorder but was subsequently diagnosed as having a pyruvate dehydrogenase deficiency. A muscle biopsy was performed and the data obtained suggested thiamine treatment which resulted in a regression of the clinical findings and a return to normal values of blood lactic and pyruvic acids. The interruption of thiamine supplementation after 1 year of treatment led to a prompt recurrence of the previous clinical and biochemical symptoms.
Biopsy↗
Reactive oxygen species as mediators of organ dysfunction caused by sepsis, acute respiratory distress syndrome, or hemorrhagic shock: potential benefits of resuscitation with Ringer's ethyl pyruvate solution.
Reactive oxygen species are reactive, partly reduced derivatives of molecular oxygen. Important reactive oxygen species in biological systems include superoxide radical anion, hydrogen peroxide, and hydroxyl radical. Peroxynitrite, is another important species in biological systems. A variety of enzymatic and non-enzymatic processes can generate reactive oxygen species in mammalian cells. An extensive body of experimental evidence from studies using animal models supports the view that reactive oxygen species are important in the pathogenesis of ischemia-reperfusion syndromes, sepsis, acute respiratory distress syndrome, and multiple organ dysfunction syndrome. This view is further supported by data from clinical studies that correlate biochemical evidence of reactive oxygen species-mediated stress with the development of acute respiratory distress syndrome or sepsis in patients. Ethyl pyruvate, a simple derivative of pyruvic acid, has been shown to be efficacious in several animal models of critical illness, and warrants further evaluation in this regard.
Adult↗
Hemodynamic effects of isoproterenol in canine endotoxin shock.
Myocardial function and peripheral hemodynamic alterations were measured through the late stages of canine endotoxin shock. 60 min postendotoxin paired animals were given infusions of either 5 ml/kg per hr of 5% dextrose or dextrose plus isoproterenol (0.25 mug/kg per min). Comparable blood lactic and pyruvic acid levels were determined, the excess lactic acid calculated, and pH values were obtained. During the initial stages the classic pattern of hemodynamic alterations was observed; an excess of lactic acid appeared and the pH decreased. Outstanding was evidence of markedly reduced myocardial function in the late stages of shock with progressive rise in left ventricular end diastolic pressure (LVEDP), low cardiac index, rise of central venous pressure, increased central blood volume, tachycardia, and declining arterial pressure. Analyses of left ventricular function curves also indicated myocardial failure. Infusion of dextrose alone failed to decrease mortality rate (10 of 18 dying), whereas the rate was significantly decreased with isoproterenol (2 of 18). Dextrose infusion did not benefit myocardial function. Isoproterenol resulted in a marked improvement in myocardial action with a significant increase in heart work associated with, yet very minor, increments of LVEDP. In addition, tachycardia subsided, peripheral resistance decreased, and the blood pressure stabilized. The prognostic value of excess lactic acid was doubtful but a progressive fall in later stages was associated with survival.
Animals↗
1H-NMR analysis of microbial-derived organic acids in primary root carious lesions and saliva.
In addition to lowered pH values, the molecular profile and concentrations of microbial-derived organic acids in carious dentin are important demineralization parameters involved in the induction, development and progression of dental caries. High-resolution proton ((1)H) NMR spectroscopy was employed to examine the organic acid status of primary root carious lesions. (1)H-NMR analysis of post-neutralized perchloric acid extracts of active carious lesions revealed that at an operating frequency of 600 MHz, the (1)H-NMR-detectable organic acid composition of carious dentin samples (mean molecular percentage content +/- standard error; the mean molecular percentage content is defined here as the mean of the concentration of each (1)H-NMR-visible organic acid/anion expressed as a percentage of total (1)H-NMR-detectable organic acid/anion level in each sample) was acetate 51 +/- 2%, formate 37 +/- 2%, lactate 5 +/- 1%, propionate 3 +/- 0.8%, pyruvate 2.4 +/- 0.3%, n-butyrate 1.2 +/- 0.2%; succinate 0.1 +/- 0.1%; iso-butyrate, n- and iso-valerate, and n- and iso-caproate (total) <0.2%. Further components detectable included alanine, glycine, choline, phosphorylcholine, trimethylamine oxide, methanol, glycolate and assorted saccharides. In view of their high dissociation constants (K(a)), our results demonstrate that formic and pyruvic acids (K(a) = 1.77 x 10(-4) and 3.20 x 10(-3) mol/dm(3), respectively) contribute substantially to the decreased pH values associated with active caries lesions (cf. lactate K(a) = 1.40 x 10(-4) mol/dm(3)), and hence the pathogenesis of primary root caries.
Aged↗
The Sulfolobus solfataricus electron donor partners of thermophilic CYP119: an unusual non-NAD(P)H-dependent cytochrome P450 system.
CYP119 from Sulfolobus solfataricus is the first well-characterized thermophilic cytochrome P450 enzyme. The endogenous substrate for this enzyme is not known but it hydroxylates lauric acid in a reaction supported by surrogate mesophilic electron donors. However, reconstitution of a high-temperature catalytic system requires identification of the normal thermophilic electron donor partners of CYP119. Here, we describe cloning, expression in Escherichia coli, and characterization of the requisite electron donor partners from S. solfataricus. One is a thermostable ferredoxin and the second a 2-oxoacid-ferredoxin oxidoreductase that utilizes pyruvic acid rather than NAD(P)H as the source of reducing equivalents. CYP119 is the only cytochrome P450 to date known to obtain electrons from a non-NAD(P)H-dependent protein. The two thermophilic partners have been used to reconstitute a catalytic system that hydroxylates lauric acid at 70 degrees C, and the optimal conditions for this system have been defined. This first high-temperature in vitro catalytic system represents an important step in the development of industrially relevant catalysts.
Amino Acid Sequence↗
Studies on the physiopathology of chronic obstructive pulmonary disease in the horse. V. Blood gas and acid-base values during exercise.
The haemoglobin concentration, the partial pressures of oxygen and carbon dioxide, the oxygen content and the pH were determined in the arterial and mixed venous blood of 5 normal and 3 horses with chronic obstructive pulmonary disease (COPD) at 3 stages of an exercise distance of 1200 m. Arterial and mixed venous samples were collected simultaneously by means of an automatic technique during the walk, trot and gallop at 0-100 m, at 500-600 m and at 1100-1200 m. The standard bicarbonate and the lactic and pyruvic acid concentrations were also determined in arterial and mixed venous blood. Highly significant changes in the mean values of PvO2. O2 content and delta a-vO2 content occurred during exercise in COPD subjects, and significant changes in PvO2 and delta a-vO2 content occurred during exercise in normal subjects. We concluded that COPD subjects compensated for respiratory dysfunction during exercise by extracting more oxygen from the blood than did normal horses. There was a highly significant correlation between the changes in standard bicarbonate and the changes in lactic acid concentration during exercise in both normal and COPD subjects. This led to the conclusion that lactic acid production was primarily, but not completely, responsible for the metabolic acidosis of exercise in horses.
Animals↗
Exercise-induced increase in glandular kallikrein activity in human plasma and its significance in peripheral glucose metabolism.
Glucose uptake into skeletal muscle of human forearm at rest and during exercise is reported to be diminished by the administration of a kallikrein (kallidinogenase) inhibitor. The present study was conducted to clarify the changes of glandular kallikrein (GK) activity in human plasma during acute exercise and its significance in peripheral glucose metabolism. 10 non-diabetic inpatients, aged 49.5 years, and 8 diabetic inpatients, aged 53.8 years, were studied. After an overnight fast, bicycle ergometer exercise test was performed for 15 min (25 W (5 min)----50 W (5 min)----75 W (5 min]. Before (basal), during (at 15 min) and after exercise (at 25 min), venous blood samples were drawn to determine plasma GK activity and glucose, serum immunoreactive insulin (IRI), C-peptide immunoreactivity (CPR), nonesterified fatty acids, pyruvate, lactate, noradrenaline and adrenaline levels. In the 1st trial, in both non-diabetics (n = 10) and diabetics (n = 8), plasma GK activity increased significantly during exercise. After the 1st trial, in 6 patients (5 non-diabetics; 1 diabetic), exercise test was repeated once after 2-4 weeks and in a diabetic patient, exercise test was repeated twice at 4-week intervals, accordingly exercise test was performed 26 times in 18 patients in total. Natural logarithmic correlation between sigma glucose level and sigma GK activity (r = 0.52, n = 26), and hyperbolic correlation between sigma glucose level and sigma IRI level (r = -0.66, n = 26) but no correlation between sigma glucose level and sigma CPR level (r = 0.17, n = 26) were found.(ABSTRACT TRUNCATED AT 250 WORDS)
Blood Glucose↗
Oxidative and glycolytic metabolism of semen components by washed guinea pig spermatozoa.
The concentration of several potentially metabolizable substances in guinea pig semen and the ability of these substances to support ATP synthesis and the motility of guinea pig sperm have been determined. Both glucose and fructose were present in high concentration in semen and were equipotent at the concentration tested in maintaining high levels of ATP and a high rate of motility. Lactic and pyruvic acids also supported a high rate of sperm motility but maintained lower levels of ATP. These constituents of guinea pig semen, as well as the metabolites alpha-glycerophosphate, succinic acid, and beta-hydroxybutyric acid, are oxidized at unusually high rates. The active oxidative metabolism of guinea pig sperm is compared with that of human sperm which is primarily glycolytic.
Adenosine Triphosphate↗
Lipid-bound saccharides in Rhizobium meliloti.
The lipid-bound saccharides formed by incubation of uridine diphosphate glucose with a particulate enzyme of Rhizobium meliloti were studied. They behaved like polyprenyl diphosphate saccharides when treated with ammonia or hot phenol, when catalytically hydrogenated, and on DEAE-cellulose chromatography. The saccharide moieties obtained after heating at pH 2 for 10 min at 100 degrees C were separated with a gel filtration column. The following compounds were detected: galactose, glucosyl beta 1-3 galactose (Tolmasky, M. E., Staneloni, R. J., Ugalde, R. A., and Leloir, L. F. (1980) ARch. Biochem. Biophys. 203, 358-364), and some octasaccharides (I). These were compared by paper electrophoresis, thin layer and paper chromatography with an octasaccharide obtained from Alcaligenes faecalis var. myxogenes strain 11 (II). Furthermore, Compounds I and II were compared with the exopolysaccharide of Rhizobium meliloti (III) by partial acid hydrolysis and methylation analysis. The results were consistent with the identity of the repeating unit of Compound III with Compounds I and II except for differences in the substituents (acetyl or succinyl). Studies on the labeling of the lipid-bound saccharides have shown that the sequence is: first, galactose and glucosyl beta 1-3 galactose, then the rest of glucose residues, and finally, the substituents (acetyl and pyruvic acid).
Chromatography, Paper↗
A teichoic acid of Nocardioides albus VKM Ac-805T cell walls.
A teichoic acid of Nocardioides albus VKM Ac-805T cell walls, a typical species of the genus Nocardioides, contains a poly(glycosylglycerol phosphate). The repeating unit of the polymer has the structure: [figure]. These units are in phosphodiester linkage at C-3 of glycerol and C-3 of beta-D-galactopyranose. beta-D-Galactopyranosyl residues are substituted at C-4 by beta-D-glucopyranose carrying a 4,6-pyruvate ketal group in S-configuration. The presence of pyruvic acid in the majority of repeating units increases the anionic properties of the polymer in comparison with most other common teichoic acids. This is the first report of the occurrence of a beta-D-galactofuranosyl residue in teichoic acids; it probably acts as a terminator of an extending chain of the polymer. The ratio of beta-D-galactopyranosyl to beta-D-galactofuranosyl units is 7:1. The polymer structure was determined by NMR spectroscopy. This type of teichoic acid structure has not been reported previously.
Actinomycetales↗
Lactate racemase. Hydroxylamine-dependent 18O exchange of the alpha-hydroxyl of lactic acid.
The lactic acid racemase (EC 5.1.2.1) derived from Clostridium butylicum catalyzes the racemization of the alpha-18O label. The proposed alpha-carbonyl intermediate for the enzyme-catalyzed reaction has been previously shown to be trapped as an enzyme-bound oxime in the presence of hydroxylamine. This report demonstrates that the formation of the inactive enzyme-bound oxime, followed by reactivation in the presence of an excess of competing free carbonyl (pyruvic acid) results in a complete loss of the alpha-18O label from an original alpha-18O-labeled lactic acid.
Clostridium↗
Cell wall teichoic acids of two Brevibacterium strains.
Structurally identical teichoic acids were detected in cell walls of two soil isolates assigned to Brevibacterium linens based on phylogenetic data. Both cell walls contain unsubstituted 1,3-poly(glycerol phosphate) and poly(glycosylglycerol phosphate). Repeating units of the latter--alpha-D-GlcpNAc-(1-->4)-beta-D-Galp-(1-->1)-Gro--are bound by phosphodiester bonds including OH-3 of galactose and OH-3 of glycerol. Some of the N-acetylglucosamine residues have 4,6-pyruvic acid acetal, amounts of the latter in the two strains being unequal. Species-specificity of the structures of teichoic acids in the genus Brevibacterium is discussed.
Brevibacterium↗
Cerebral blood flow and exchange of oxygen, glucose ketone bodies, lactate, pyruvate and amino acids in anesthetized children.
Cerebral blood flow (CBF) and cerebral av differences of oxygen, glucose, 3-hydroxybutyrate, acetoacetate, lactate, pyruvate and amino acids were measured in anaesthetized children before elective surgery in order to study possible age-dependent variations. CBF was measured in 70 children, aged 11 days to 15 years. Cerebral av differences were studied in approximately 50% of the subjects. Mean values were: CBF 0.65 ml x g-1 x min-1, cerebral exchange in nmoles x g-1 x min-1; 1348, glucose 248, acetoacetate 12,3-hydroxybutyrate 34 (uptake), lactate-48, pyruvate-8 (release). No net exchange of amino acids was found with the exception of histidine (uptake). Neither CBF nor the cerebral exchange of oxygen and circulating substrates showed any correlation to age within the group. Compared with adults anesthetized by the same technique (barbiturate induction, nitrous oxide-oxygen relaxant) the children had a slightly higher mean CBF, while the cerebral uptake of oxygen and glucose were equal to values in adults. The cerebral uptake of ketone bodies was higher in children than reported values in adults investigated in the awake state after comparable periods of fasting.
Acetoacetates↗