Antagonism in the simultaneous oxidation of D-amino acids by D-amino acid oxidase.
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This study compared the NADH- and NADPH-supported p-nitrophenetole (NP) O-deethylase, ethylmorphine (EM) O-deethylase and EM N-demethylase activities of rat hepatic microsomes with respect to dioxygen requirement, inhibition by carbon monoxide, inhibition by classical inhibitors of cytochrome P-450 systems, and the involvement of NADH-cytochrome b5, cytochrome b5 reductase and NADPH-cytochrome P-450 reductase. The results generated the following conclusions and speculations: NADH- and NADPH-supported O-deethylations of NP involve different P-450 hemoproteins. This conclusion was based largely on the observations that the NADPH-supported reaction was inhibited by carbon monoxide and cyanide (5 mM), whereas the NADH-supported reaction was not; the NADH-supported reaction required a relatively high pO2 for maximal activity, whereas the NADPH-supported reaction did not, and the NADPH-supported reaction was depressed in microsomes from rats that had been administered Co2+, Mn2+, allylisopropylacetamide (AIA) or polyriboinosinic acid X polyribocytidylic acid (poly IC), whereas the NADH-supported reaction was not. However, the NADH- and NADPH-supported reactions shared some common features: both were strongly inhibited by alpha-naphthoflavone and weakly inhibited by 2-diethylaminoethyl 2,2-diphenyl valerate HCI (SKF 525-A), both were destroyed by linoleic acid hydroperoxide, and both were induced by 3-methylcholanthrene (MC) and phenobarbital. The use of antibodies against NADPH-cytochrome P-450 reductase, NADH-cytochrome b5 reductase and cytochrome b5 demonstrated that both the NADH- and the NADPH-supported reactions depend on established components of cytochrome P-450 systems. The P-450 hemoproteins involved primarily in both the NADH- and NADPH-supported deethylation of NP are the P1-450 type, i.e. they are markedly induced by MC and inhibited by alpha-napthoflavone. The NADH- and NADPH-supported O-deethylations of NP involve separate electron transfer systems.(ABSTRACT TRUNCATED AT 400 WORDS)
The intermediate, medial part of the hyperstriatum ventrale (IMHV) is a region of the avian forebrain which is known to be essential for early learning in the domestic chick. The IMHV in an in vitro slice preparation displays two forms of synaptic plasticity. The incidence of both varies with age and is maximal between 3 and 5 days post-hatch. Since NMDA receptors are critical for at least one of these plasticities, we have investigated the relationship between age and the contribution of NMDA receptors to the field response evoked by local, low-frequency stimulation and have found that the magnitude of the NMDA-dependent component of the response varies with age peaking between 3 and 5 days post-hatch. Spontaneous neural activity, recorded intracellularly, can be completely and reversibly silenced by NMDA receptor blockade and the incidence of spontaneous activity also varies with age, peaking between 3 and 5 days. These results suggest that the IMHV contains NMDA receptors which can be activated near resting membrane potential. Either the efficiency or the numbers of these receptors is maximal at a specific point in development and their peak activity coincides with a peak in synaptic plasticity. These characteristics are similar to those reported for young mammals.
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A reliable and practicable method for the determination of short chain fatty acids in plasma and brain tissue is presented. The sample preparation by partition chromatography on silicic acid allows subsequently a quantitation of short chain fatty acids without interference by methylmalonic acid, or other more polar compounds. With the gas-chromatographic system 2-methylbutyrate is separated from isovalerate. Reference values are given for plasma. The system is also useful in combination with mass spectrometry.
A method is described in which ammonia chemical ionization gas chromatography-mass spectrometry was utilized in the selected ion monitoring mode to provide an accurate, selective approach to the quantification in amniotic fluid of a number of hydroxylated organic acids derived from the metabolism of the branched-chain amino acids. 2-Hydroxy-n-caproic acid was employed as an internal standard and the hydroxy acids were isolated from amniotic fluid by liquid partition chromatography and the trimethylsilyl derivatives were quantified. Normal values have been obtained for 2-hydroxyisovaleric acid, the sum of 2-hydroxyisocaproic acid and 2-hydroxy-3-methylvaleric acid, 2-methyl-3-hydroxybutyric acid, 3-hydroxyisovaleric acid and 2-ethyl-3-hydroxypropionic acid. The method also provides data on the concentration of methylmalonic acid. The concentration of 2-hydroxyisovaleric acid was not useful in the prenatal diagnosis of a fetus with maple syrup urine disease. Elevated concentrations of 2-methyl-3-hydroxybutyric acid as well as methylmalonic acid were found in the amniotic fluid of two fetuses with methylmalonic acidemia.
The efficacy of clofibrate (CPIB) and nicotinic acid (NA) in the treatment of type III hyperlipoproteinemia was evaluated in 5 male subjects in a randomized cross-over study with clofibrate 1 g b.i.d. and NA 3 g/day (given either b.i.d. or t.i.d.). Following a baseline period of 6 weeks, each drug was given for 12 weeks with samples for lipid and lipoprotein determinations obtained at 6, 9, and 12 weeks. Both clofibrate and NA resulted in a significant reduction from baseline of total cholesterol (23% and 28%), VLDL cholesterol (49% and 56%), total triglycerides (40% and 43%), and VLDL triglycerides (46% and 48%), as well as a significant increase in HDL cholesterol (22% and 28%) and HDL/LDL ratio (31% and 62%). The HDL/LDL ratio was higher on NA than clofibrate (0.47 +/- 0.19 vs. 0.38 +/- 0.09, P less than 0.05). Four subjects were continued in the study and treated sequentially with NA 3.0 g/day (alternate to the previous schedule) and gemfibrozil 1.2 g/d in divided doses. Each of the 4 regimens resulted in a significant change from baseline of each of the measured lipid and lipoprotein determinations except LDL cholesterol. Comparison among the treatment regimens revealed no differences except for significantly higher HDL cholesterol and HDL/LDL ratio with NA given t.i.d.
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The present work deals with the kinetics of hydrolysis of the acyl derivatives of stobadine, an originally synthesized potential antiarrhythmic and antihypoxic drug, which was found to have also an excellent scavenging effect on reactive oxygen species. The acyl derivatives of stobadine, which possess high lipophilicity, represent model blood-brain barrier penetrating agents. It is assumed that the acyl derivatives of stobadine may act as prodrugs which are hydrolysed in different biological tissues to release the active drug. The decomposition of three acyl derivatives of stobadine was studied in acidic, basic and neutral buffer solutions at constant ionic strength (0.1 mol/L) at 25 degrees and 70 degrees C using UV spectrophotometric method. The pseudo first-order rate constants and the pH-rate profile for the degradation of acetyl-, valeroyl- and nicotinoyl-derivatives of stobadine were determined. Confirmation that stobadine was the first degradation product was provided by thin-layer chromatography.
Positive-ion thermospray liquid chromatography-mass spectrometry (TSP-LC-MS) is used to detect organic acids via the direct injection of untreated urine from newborns and infants. Two methods are reported for the separation of organic acids. The separation of urinary organic acids is effected in either an acidic, pH 2.5 sulfuric acid, or a non-acidic, 0.05 M ammonium acetate, pH 6.8, mobile phase. Use of pH 2.5 sulfuric acid and an HPX-87H organic acid column produces better separation but has less sensitivity than the use of 0.05 M ammonium acetate, pH 6.8 and a C18 column. Positive ion TSP-LC-MS has been used to detect methylmalonic aciduria, 3-hydroxy-3-methylglutaric aciduria, propionic aciduria, isovaleric aciduria and argininosuccinic aciduria.
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