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[Reference values of urinary orotic acid in a healthy Tunisian population].

BACKGROUND: Orotic acid (OA) is an intermediary metabolite of pyrimidine synthesis. An elevation of urinary orotic acid excretion has been described in congenital defect of the urea cycle enzymes and in primary orotic aciduria. Several techniques have been used to measure OA and many reference values are published without considering age and sex. POPULATION AND METHODS: The reference values of urinary OA excretion, expressed in mumol/mmol of creatinine, are reported in a healthy Tunisan population using a colorimetric method. The study included 20 men and 20 women (age = 31 +/- 11 years) and 30 children aged from 3 days to 8 years. RESULTS: There was a significant increase (P < 0.01) of urinary OA excretion in women (4.38 +/- 1.35) compared to men (3.26 +/- 0.80) and of children (5.03 +/- 1.14) compared to adults (3.82 +/- 1.24). Urinary OA excretion was significantly higher among children aged less than 1 year compared to older children. CONCLUSION: It is necessary for every laboratory practicing this kind of exploration to have its own norms which depend on both age and sex.

Adult↗

Dietary orotic acid affects antioxidant enzyme mRNA levels and oxidative damage to lipids and proteins in rat liver.

We investigated the effects of the dietary addition of orotic acid on liver antioxidant enzymes, mRNA levels of these enzymes, and peroxidative products by comparing casein with soy protein as the source of dietary protein. Rats fed the casein diet accumulated more liver lipids than those fed the soy protein diet when orotic acid was added. The addition of orotic acid lowered both the activity of liver Cu, Zn-superoxide dismutase and the level of Cu, Zn-superoxide dismutase mRNA. The addition of orotic acid led to a significant increase in the contents of conjugated dienes and protein carbonyls in the liver. In addition, dietary soy protein protected the increase in the levels of lipids and proteins peroxide induced by orotic acid. The addition of orotic acid to the casein diet increased the activities of both serum ornithine carbamoyltransferase and alanine aminotransferase. Thus, liver damage might result from the increased superoxide anion due to the decrease in the activity of hepatic superoxide dismutase, as well as increase in the production of hepatic peroxidative products in rats fed the casein diet with orotic acid.

Journal Article↗

Determination of orotic acid, uric acid, and creatinine in milk by liquid chromatography.

A simultaneous determination of orotic acid, uric acid, and creatinine in milk is described. Following deproteinization, the sample was analyzed by reversed-phase liquid chromatography, using a highly aqueous cationic ion-pair eluent and photodiode array UV detection. In view of their potential dietary significance, the validated method was applied to survey the influence of species, season, and lactation on their contribution to the nonprotein nitrogen pool in milk. Mature bovine milk contained orotic acid, uric acid, and creatinine in the range of 30-70, 9-24, and 6-12 microg/mL, respectively. Although uric acid and creatinine were present in all milks, orotic acid was essentially absent in nonruminant milks. In contrast to urate and creatinine, expression of orotic acid in bovine milk was strongly dependent on stage of early lactation. The co-existence in mammalian milks of related nucleoside and nucleotide components was also determined.

Animals↗

Effect of orotic acid on liver glycogen of different animal species.

The effect of orotic acid on the liver glycogen content in the mice, frogs and catfish was studied. It was observed that the orotic acid significantly increases the glycogen content in the liver of mice and catfish as it does in rats. On the other hand it causes a fall of the glycogen level in frogs in experiments made both in autumn and spring. This effect was modified by amino acids administered together with orotic acid.

Animals↗

Cycloheximide sensitivity of orotic acid biosynthesis induced by ammonia and glycine administration.

Administration of either ammonia or glycine to both rats and mice results in an increased synthesis in the liver and urinary excretion of orotic acid. The two most relevant observations obtained are that carbamoyl phosphate synthesized inside the mitochondria is involved in the increased synthesis of orotic acid and that this latter process is almost completely abolished by cycloheximide and actinomycin D, inhibitors of protein and RNA synthesis. Orotic acid synthesis could be controlled by an induction-suppression mechanism. Inhibition of synthesis of excess orotic acid brought about by N-(phosphonacetyl)-L-aspartic acid but not by acivicin, suggests that glutamine-dependent cytosolic synthesis of carbamoyl phosphate, is not involved. Administration of ornithine together with glycine completely suppressed the synthesis of orotic acid, but promoted a twofold increase of urea excretion. The concentration of ornithine rather than that of carbamoyl phosphate or the activity of the enzymes involved, may represent a limiting factor controlling both the flux of ammonia in the urea cycle and the availability of mitochondrial carbamoyl phosphate for orotic acid synthesis. Two enzymes have been found to be induced by glycine: ornithine decarboxylase and aspartate transcarbamoylase (aspartate carbamoyltransferase). Both enzymes may contribute to the increase in orotic acid synthesis, aspartate transcarbamoylase more directly and ornithine decarboxylase by lowering the ornithine concentration. Ornithine decarboxylase activity was completely suppressed but that of aspartate transcarbamoylase was further increased by cycloheximide treatment. Inhibition of orotic acid biosynthesis by cycloheximide appears to be the result of a decreased availability in the cytosol of carbamoyl phosphate synthesized inside the mitochondria.

Ammonium Chloride↗

Determination of orotic acid in children's urine.

A reliable method is described for the determination of urinary orotic acid. The orotic acid is separated quickly and easily from the urine by anion exchange chromatography. Reference values are reported for children. The method is suitable for the differentiation of those inherited metabolic defects that lead to hyperammonemia.

Adolescent↗

Reduction of beta-oxidation capacity of rat liver mitochondria by feeding orotic acid.

Rats were maintained on fat-free high carbohydrate diets either with or without orotic acid (1%, w/w), pantethine (1%, w/w), adenine (0.25%, w/w), and/or p-chlorophenoxyisobutyrate (0.25%, w/w). Oxidation of fatty acid by liver mitochondria was inhibited to less than half that of the control after administration of orotic acid. Activities of acyl-CoA dehydrogenases were markedly decreased by orotic acid administration, but the following enzyme activities were not, or only slightly decreased: acyl-CoA synthetase, carnitine acyltransferases, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase and 3-ketoacyl-CoA thiolase. Simultaneous addition of pantethine in the orotic acid-containing diet prevented induction of fatty liver. It also prevented decreases in fatty acid oxidation capacity and acyl-CoA dehydrogenase activity. Introduction of adenine or p-chlorophenoxyisobutyrate, which reverse orotic acid-induced fatty liver, reversed oxidation and acyl-CoA dehydrogenase activities to control levels. The oxidation capacity of the peroxisomal system remained unchanged after administration of orotic acid.

Acyl-CoA Dehydrogenases↗