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Urinary excretion of purines in sheep with experimental orotic aciduria.

Urinary excretion of uric acid, hypoxanthine, allantoin, and urea was measured in sheep (Polish Merino, about 20 kg b.w.) with experimental orotic aciduria. The 240 min infusion of 6-azauridine solution into the jugular vein induced a highly significant increase of urinary orotic acid, uric acid and hypoxanthine excretion. No differences were found in relation to excretion of allantoin and urea in examined sheep. It was calculated that renal clearance of uric acid and hypoxanthine increased significantly in response to 6-azauridine infusion. Intravenous infusion of sodium orotate evoked a highly significant elevation of renal urate clearance. No significant change in renal urea clearance was observed. The data suggest that competition between the renal transport of orotate and actively transported purine compounds (uric acid and hypoxanthine) occurs.

Allantoin↗

Determination of organic acids, sugars, diacetyl, and acetoin in cheese by high-performance liquid chromatography.

Sugars (lactose, glucose, and galactose), nonvolatile acids (citric, orotic, piruvic, lactic, ossalic, and hippuric), some free fatty acids (formic, acetic, propionic, butyric, isobutyric, valeric, and isovaleric), diacetyl, and acetoin were separated on an Aminex HPX-87H column using a simple isocratic HPLC method and identified by retention times with ultraviolet and refractive index detectors. With the proposed technique it is possible to evaluate the development of microbial fermentations and, at the same time, degree of cheese ripening.

Acetoin↗

Effect of dietary nucleotides and orotate on the blood levels of prostacyclin (PGI2) and thromboxane (TXA2) in the weanling rat.

Dietary nucleotides affect the maintenance of immune responses, tissue repair and polyunsaturated fatty acid metabolism. Orotate, a pyrimidine nucleotide precursor, induces fatty livers by impairing VLDL hepatic secretion. The aim of this study was to evaluate the changes in the blood levels of fatty acids and prostacyclin (PGI2) and thromboxane (TXA2) in the weanling rat caused by the dietary intake of nucleotides and orotate. Three groups of rats at weaning were fed a control diet, an orotate supplemented diet (O-50) and a nucleotide supplemented diet (N-50) during 4 weeks, respectively. Absolute values of plasma polyunsaturated fatty acids greater than 18 carbon atoms of the n-6 and n-3 series were increased in the N-50 group and decreased in O-50 with regard to the control. However, the relative fatty acid composition of plasma lipid fractions was mostly unaffected. Plasma 6-keto-PGF1 alpha showed a trend to be increased in N-50 and serum TXB2 was significantly increased in that group. Both eicosanoids were unchanged by dietary orotate intake. These results may be explained because of the increased plasma 20:4n-6 found in rats fed a supplemented nucleotide diet. Thus, nucleotides present in foods appear to modulate PUFA conversion and eicosanoids synthesis in early life.

6-Ketoprostaglandin F1 alpha↗

Pyrimidine nucleobase ligands of orotate phosphoribosyltransferase from Toxoplasma gondii.

Sixty-seven pyrimidine nucleobase analogues were evaluated as ligands of Toxoplasma gondii orotate phosphoribosyltransferase (OPRTase, EC 2.4.2.10) by measuring their ability to inhibit this enzyme in vitro. Apparent Ki values were determined for compounds that inhibited T. gondii OPRTase by greater than 20% at a concentration of 400 microM. 1-Deazaorotic acid (0.47 microM) and 5-azaorotic acid (2.1 microM) were found to bind better (8.3- and 1.9-fold, respectively) to T. gondii OPRTase than orotic acid, the natural substrate of the enzyme. Based on these results, a structure-activity relationship of ligand binding to OPRTase was formulated using uracil, barbituric acid, and orotic acid as reference compounds. It was concluded that the following structural features of pyrimidine nucleobase analogues were required or strongly preferred for binding: (i) an endocyclic pyridine-type nitrogen or methine at the 1-position; (ii) exocyclic oxo groups at the 2- and 4-positions; (iii) a protonated endocyclic pyridine-type nitrogen at the 3-position; (iv) an endocyclic pyridine-type nitrogen or methine at the 5-position; (v) an exocyclic hydrogen or fluorine at the 5-position; (vi) an endocyclic pyridine-type nitrogen or methine at the 6-position; and (vii) an exocyclic negatively charged or electron-withdrawing group at the 6-position. A comparison of the results from the present study with those from a previous study on mammalian OPRTase [Niedzwicki et al., Biochem Pharmacol 33: 2383-2395, 1984] identified four compounds (6-chlorouracil, 5-azaorotic acid, 1-deazaorotic acid, and 6-iodouracil) that may bind selectively to T. gondii OPRTase.

Animals↗

Ribonucleic acid synthesis in porphyric rat liver induced by 3,5-dicarbethoxy-1,4-dihydrocollidine.

1. Administration of 3,5-dicarbethoxy-1,4-dihydrocollidine to rats caused a marked increase in the activity of delta-aminolaevulinic acid synthetase as well as a slight net increase in RNA in the livers. 2. 3,5-Dicarbethoxy-1,4-dihydrocollidine primarily stimulated the synthesis of RNA in the nucleus of the liver cell. 3. The decrease in RNA synthesis after administration of this drug resulted in a rapid decrease in the activity of this induced enzyme. This was also confirmed by treatment with actinomycin D. 4. From kinetic experiments on synthesis of RNA and delta-aminolaevulinic acid synthetase in vivo the induction mechanism of this enzyme was discussed.

5-Aminolevulinate Synthetase↗