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Electrochemically treated glassy carbon electrode for amperometric detection in high-performance liquid chromatography.

The application of an electrochemically pre-treated glassy carbon electrode for the amperometric detection of electroactive components, such as tyrosine and oxipurinol, in biological samples was studied in order to demonstrate the usefulness of a pre-anodized electrode in high-performance liquid chromatography. The electrochemical pre-treatment was carried out in 0.2 M phosphate buffer (potassium dihydrogenphosphate-potassium hydroxide, pH 6.5) at 1900 mV vs. Ag/AgCl for 2 min. The pre-anodized electrode response for the oxidation of lactic acid and pyruvic acid was also studied. The electrochemical treatment enhanced and stabilized the electrode response to the oxidation of tyrosine and both acids.

Chromatography, High Pressure Liquid↗

Effect of octreotide acetate on the plasma concentration and urinary excretion of uridine and purine bases.

To determine the effect of octreotide acetate on urinary excretion of uric acid and plasma concentration of uridine, we subcutaneously administered octreotide acetate (1 microg/kg of body weight) to 5 healthy subjects. Ninety minutes after administration, octreotide acetate increased the plasma concentration of uridine by 15% and decreased the plasma concentration of glucagon by 24% and that of insulin to below the detection limits. In addition, octreotide acetate decreased the urinary excretion of uric acid, sodium, and chloride by 60%, 40%, and 38%, respectively, at 1 hour after administration. However, octreotide acetate did not affect the concentrations of hypoxanthine, xanthine, uric acid, cyclic AMP in plasma, lactic acid and pyruvic acid in blood, urinary excretion of hypoxanthine and xanthine, or creatinine clearance. From these results, we speculated that octreotide acetate decreases the urinary excretion of uric acid by decreasing the concentration of glucagon and/or urinary excretion of sodium, and increases the plasma concentration of uridine via decreased concentrations of glucagon and insulin.

Adult↗

[Protective action of sophoridine on acute myocardial infarction in anesthetized dogs].

The effects of sophoridine on acute myocardial infarction induced by ligation of left anterior descending coronary artery in 16 anesthetized and open-chest dogs were studied. Treatment with sophoridine 20 mg.kg-1 iv reduced the size of myocardial infarct 6 h after ligation and lowered the elevated plasma levels of lactic acid and pyruvic acid 15, 30 and 60 min after ligation. Sophoridine also lowered the activity of serum CPK 60 and 120 min after ligation and reduced TXB2 and increased 6-keto-PGF1 alpha, hence the metabolites of PGI2/TXA2 ratio after ligation was increased. Electron microscopy showed that sophoridine protected mitochondria of ischemic cells. These results indicated that sophoridine showed protective effects on acute myocardial infarction. The beneficial effect on PGI2/TXA2 and mitochondria seems to contribute its protective action.

6-Ketoprostaglandin F1 alpha↗

Blood carbohydrates and phosalone poisoning in Rana tigrina (Daudin).

In Rana tigrina, blood levels of total carbohydrates, lactic acid and pyruvic acid were determined in relation to allometric growth in both sexes. The blood carbohydrates content increased in relation to growth and maturity of gonads. Treatment by phosalone resulted in the accumulation of lactic acid in blood suggesting that this organophosphate compound inhibits the normal oxidative metabolism.

Animals↗

Purification and characterization of indolepyruvate decarboxylase. A novel enzyme for indole-3-acetic acid biosynthesis in Enterobacter cloacae.

Indolepyruvate decarboxylase, a key enzyme for indole-3-acetic acid biosynthesis, was found in extracts of Enterobacter cloacae. The enzyme catalyzes the decarboxylation of indole-3-pyruvic acid to yield indole-3-acetaldehyde and carbon dioxide. The enzyme was purified to apparent homogeneity from Escherichia coli cells harboring the genetic locus for this enzyme obtained from E. cloacae. The results of gel filtration experiments showed that indolepyruvate decarboxylase is a tetramer with an M(r) of 240,000. In the absence of thiamine pyrophosphate and Mg2+, the active tetramers dissociate into inactive monomers and dimers. However, the addition of thiamine pyrophosphate and Mg2+ to the inactive monomers and dimers results in the formation of active tetramers. These results indicate that the thiamine pyrophosphate-Mg2+ complex functions in the formation of the tetramer, which is the enzymatically active holoenzyme. The enzyme exhibited decarboxylase activity with indole-3-pyruvic acid and pyruvic acid as substrates, but no decarboxylase activity was apparent with L-tryptophan, indole-3-lactic acid, beta-phenylpyruvic acid, oxalic acid, oxaloacetic acid, and acetoacetic acid. The Km values for indole-3-pyruvic acid and pyruvic acid were 15 microM and 2.5 mM, respectively. These results indicate that indole-3-acetic acid biosynthesis in E. cloacae is mediated by indolepyruvate decarboxylase, which has a high specificity and affinity for indole-3-pyruvic acid.

Carboxy-Lyases↗

Functional activity of a monocarboxylate transporter, MCT1, in the human retinal pigmented epithelium cell line, ARPE-19.

The purpose of this study was to identify and characterize the functional activity of monocarboxylic acid transporter 1 (MCT1) on the human retinal pigmented epithelium (RPE) cell line, ARPE-19, and to evaluate whether the cell line can function as an in vitro screening tool for intravitreally administered drugs/prodrugs targeted to the MCT1 expressed in RPE. Uptake studies were carried out at 37 degrees C, for 30 s, with ARPE-19 cells. [(14)C]l-Lactic acid was selected as a substrate for this transporter. Uptake of [(14)C]L-lactic acid by ARPE-19 cells was found to exhibit saturable kinetics (K(m) = 3.1 +/- 0.6 mM and V(max) = 63.1 +/- 4.1 pmol/min/mg of protein). Monocarboxylic acids, such as benzoic acid, salicylic acid, and pyruvic acid, inhibited the uptake of [(14)C]L-lactic acid whereas di- and tricarboxylic acids, such as phthalic, succinic, and citric acids, did not demonstrate any inhibitory effect. Uptake was stereospecific where D-lactic acid was less effective in inhibiting [(14)C]L-lactic acid uptake than unlabeled L-lactic acid. ELISA indicated the expression of only MCT1, MCT4, and MCT8 isoforms by ARPE-19 cells. Increase in [(14)C]L-lactic acid uptake was observed as the uptake medium pH was lowered from 7.4 to 5.0. Moreover, inhibition of [(14)C]L-lactic acid uptake was observed in the presence of the protonophore 2,4-dinitrophenol. Uptake was significantly decreased in the presence of sodium azide, ouabain, p-chloromercuribenzoic acid (pCMBA), N-ethylmaleamide, dithiothreitol, and p-chloromercuribenzene sulfonate (pCMBS). However, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and L-thyroxine did not inhibit [(14)C]L-lactic acid. RT-PCR studies and sequence analysis of the PCR product confirmed the expression of MCT1 by ARPE-19 cells. Our results indicate that MCT1 is functionally active and is the only MCT isoform involved in the apical uptake of monocarboxylates by ARPE-19 cells. This cell line may thus be used as an effective screening tool for intravitreally administered drugs/prodrugs targeted toward MCT1 expressed on the RPE.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Identification of lactate. Analytical methods of pharmacopoeias with DBH in respect to environmental and economical concern, Part 4.

The identification of lactate according to Ph. Eur. 1997 and DAB 2000 uses the oxidation of lactic acid to pyruvic acid by boiling with bromine water in sulphuric acid. Acetaldehyde arising by decarboxylation is detected according to Legal applying a time consuming and troublesome procedure. 1,3-Dibromo-5,5-dimethylhydantoin (DBH) as well as potassium bromate can replace elemental bromine. Lactic acid and all lactates of Ph. Eur. 1997 and DAB 2000 can be identified better and faster using lactate oxidase (LOD, test strip system Accusport). According to DAB 2000 the base of ethacridine lactate has to be separated. This is no longer necessary, if an enzymatic identification is applied.

Bromates↗

Changes in cerebral glycogenolysis and related enzymes in diazinon treated hyperglycaemic animals.

Effect of diazinon (10,20 and 40 mg/kg, i.p.) on the level of blood glucose in rats was investigated. Hyperglycaemia peaked 2 h after i.p. treatment with 40 mg/kg diazinon. The cerebral acetylcholinesterase activity was significantly reduced. The blood level of pyruvic acid was unchanged while that of lactic acid was significantly increased. Convulsions and biochemical changes caused by diazinon (40 mg/kg) were prevented by diazepam injected immediately after diazinon. In diazinon-treated hyperglycaemic animals, the glycogen content of the brain was depleted, the activities of glycogen phosphorylase, phosphoglucomutase and hexokinase were significantly increased and the activity of glucose-6-phosphatase remained unchanged. Lactate dehydrogenase activity was also increased by treatment with diazinon. The induced changes may compensate for the energy requirement of stimulatory effects caused by diazinon.

Animals↗

Corneal damage by half mustard (2-chloroethyl ethyl sulfide, CEES) in vitro preventive studies: a histologic and electron microscopic evaluation.

The effect of half-mustard (2-chloroethyl ethyl sulfide, CEES) on the morphology and ultrastructure of the cornea has been studied in vitro. Extensive necrotic changes were observed histologically as well as electron microscopically. The outer layer of corneal epithelium was observed to undergo vacuolization and globulization prior to its denudation. The epithelium becomes separated from the Bowman's membrane. These necrotic changes are prevented from taking place in the presence of a mixture of taurine, pyruvic acid, alpha-keto glutaric acid and pantothenic acid suggesting the use of this mixture in the prevention of mustard damage.

Animals↗

[Hormonal and metabolic criteria of the adequacy of anesthesia during surgery of nephroblastoma in children].

The data are presented on changes in hypophyseal-adrenal system and carbohydrate metabolism during transperitoneal nephrectomy in 42 children aged 1 to 15 years with stage II-III tumors. In 22 patients surgery was performed under combined neuroleptanalgesia with halothane for induction and maintenance of anesthesia. In 20 patients halothane was replaced by diazepam. Glucose, insulin, ACTH, CTH, hydrocortisone, lactic and pyruvic acid levels, total activity of LDG, its isoenzymes and cAMP have been assessed at different stages of anesthesia and surgery.

Adolescent↗

Effect of storage temperature and pyruvate on kinetics of anthocyanin degradation, vitisin A derivative formation, and color characteristics of model solutions.

The formation of vitisin A, an anthocyanin formed naturally in small quantities in maturing port wines, was studied in model wine solutions at several storage temperatures (10, 15, 20, and 32 degrees C). Vitisin A was formed through the interaction between malvidin 3-glucoside and pyruvic acid, Acylated forms of vitisin A, having the 6-position of the sugar acylated with acetic acid (3-acetylvitisin A) and p-coumaric acid (3-p-coumarylvitisin A), were also formed through the interaction between pyruvic acid and malvidin 3-acetylglucoside and malvidin 3-p-coumarylglucoside, respectively. A maximum degradation of the anthocyanins was obtained at higher temperatures, and it followed a first-order kinetics both with and without pyruvic acid in the solution. Whereas at low temperatures (10 and 15 degrees C) the presence of pyruvic acid accelerated the kinetic reaction, at higher temperatures (20 and 32 degrees C) it decreased it. The activation energy values for the degradation of the three anthocyanins in model solutions without and with pyruvic acid were not significantly different from each other. At low temperatures the highest concentrations of vitisin A compounds were obtained. All solutions showed a decrease in L value, indicating that all solutions became darker. This change increased with increasing temperature. All model solutions increased in the hue angle, indicating that the solutions changed from a bluish-red to an orange-red or even brownish-red color. Samples without pyruvic acid remained lighter and became browner than those with pyruvic acid. A good correlation between the amount of vitisin A in the solution and hue angle was found, indicating that vitisin A may contribute the orange-red of solutions, compared to the browner control.

Acetylation↗

Mechanisms for plasma-mediated activation of human blood cell aldehyde dehydrogenase.

Aldehyde dehydrogenase (ALDH; EC 1.2.1.3) activity assays were carried out on isolated human blood cells in phosphate-buffered saline (PBS) and in PBS mixed with human plasma. In assays with intact erythrocytes or sonicated leukocytes, the presence of 50% (v/v) or greater of plasma in the reaction mixtures produced a 2-fold increase in the rate of aldehyde oxidation. In corresponding assays with sonicated erythrocyte samples, the ALDH activity was enhanced on an average 1.5-fold, whereas a slight decrease was observed in assays with intact leukocytes. The ALDH inhibitor disulfiram almost completely abolished the enzyme activity both in the absence and presence of plasma. In assays with sonicated leukocytes, the activation effect could be antagonized by EDTA, indicating that it was caused largely by divalent cations. With sonicated erythrocytes, a significantly reduced ALDH activity was found only with the highest concentration of EDTA tested, and since a similar reduction was obtained also when plasma was omitted, the plasma-mediated activation of erythrocyte ALDH was suggested to be due to a different mechanism. After separation of plasma by gel filtration, an active fraction was identified by GC-MS and 1H-NMR to contain pyruvic acid, lactic acid and glucose. When tested at physiological plasma concentrations, pyruvic acid caused an increase in erythrocyte ALDH activity similar to that obtained with plasma, while lactic acid and glucose did not. Pyruvic acid did not activate the leukocyte ALDH. Based on these results, it is indicated that the plasma-mediated activation of erythrocyte ALDH is due to pyruvic acid, which reoxidizes NADH via lactate dehydrogenase (EC 1.1.1.27) and, thereby, increases the rate of dissociation of NADH from the terminal enzyme-NADH complex, the rate-limiting step in the ALDH pathway.

Aldehyde Dehydrogenase↗

Evidence for oligomer formation in clouds: reactions of isoprene oxidation products.

Electrospray ionization mass spectrometry (ESI-MS) was used to investigate product formation in laboratory experiments designed to study secondary organic aerosol (SOA) formation in clouds. It has been proposed that water soluble aldehydes derived from aromatics and alkenes, including isoprene, oxidize further in cloud droplets forming organic acids and, upon droplet evaporation, SOA. Pyruvic acid is an important aqueous-phase intermediate. Time series samples from photochemical batch aqueous phase reactions of pyruvic acid and hydrogen peroxide were analyzed for product formation. In addition to the monomers predicted by the reaction scheme, products consistent with an oligomer system were found when pyruvic acid and OH radical were both present. No evidence of oligomer formation was found in a standard mix composed of pyruvic, glyoxylic, and oxalic acids prepared in the same matrix as the samples analyzed using the same instrument conditions. The distribution of high molecular weight products is consistent with oligomers composed of the mono-, oxo-, and di-carboxylic acids expected from the proposed reaction scheme.

Aldehydes↗