[Identification of antioxidant molecules by thin-layer nanochromatography].
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Biomedical subjects
Publications and source records attributed to J Alary.
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A field study of fluoride pollution and of its consequences was made over a period of five years in the vicinity of an alumina reduction plant. This study, based upon the use of a static soda impregnated paper sampler, shows that the results obtained are in good agreement with the atmospheric fluoride concentrations obtained with dynamic samplers, especially when fluoride is present in gaseous form (HF). The results suggest that vertically mounted paper samplers are less sensitive to the collection of particulate fluorides. Collection rate is strongly influenced and increases with speed. If this parameter is known, an accurate estimation of the absolute atmospheric concentration is possible; the values shown by this technique are in good correlation with the value (annual mean) of pasture fluoride content. As a correlation exists between pasture fluoride content and the symptoms shown by cattle, the static filter samplers can predict disease in the case of slow chronic fluorosis. The field study agrees with earlier laboratory experiments and shows the validity of these simple and inexpensive types of exposure methods.
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4-Hydroxy-2-nonenal (HNE), an aldehyde end product of lipid peroxidation in biological systems, is capable of producing a range of powerful biological effects. Despite its biological relevance, the metabolic fate of this aldehyde is unknown in vivo. This study examines the urinary excretion of HNE in the rat and the nature of metabolites formed. Following iv administration of [3H]HNE, the majority of the dose appeared in urine (67.1% after 48 h). The radio-HPLC metabolic profile showed that no unchanged parent compound was detected in urine whereas at least four metabolites were present, most of them corresponding to mercapturic acid conjugates. Two major pathways were involved in the biotransformation of HNE in vivo: (i) reduction/oxidation of the aldehyde group, and (ii) conjugation to endogenous glutathione leading to mercapturic acid conjugates in urine. These end products were isolated by HPLC and identified by mass spectrometry as HNE mercapturic acid, 1,4-dihydroxynonene mercapturic acid, 4-hydroxynonenoic mercapturic acid, and the corresponding lactone.
Chloramphenicol (CP) has been implicated as, although not proven to be, a causative agent of aplastic anemia in humans. Recent studies from our laboratory have presented evidence that CP-oxamylethanolamine was an end product of CP biotransformation in birds. Because this novel metabolic pathway has never been reported in other species, we have now expanded these investigations to rat and humans. [3H]CP was administered po (10 mg/kg) to adult male Wistar rats and to a human volunteer. Urine was collected and analyzed by HPLC and GC-MS for CP metabolite determination. In rat, the two most important metabolites in 0-24 h urine were CP-base and CP-acetylarylamine which together accounted for about 50% of the ingested radioactivity. The remainder was due to unchanged CP, CP-oxamic acid, CP-alcohol, CP-glucuronide, and CP-oxamylethanolamine. The presence of these end products was also demonstrated in man. CP-oxamylethanolamine represented 0.74% and 1.37% of the ingested radioactivity in rat and human urine samples, respectively. CP-oxamylethanolamine formation was confirmed in vitro with isolated rat hepatocytes, suggesting the involvement of liver in the production of this metabolite. The origin of CP-oxamylethanolamine has been investigated with the use of hepatic liver microsomes from phenobarbital-treated rats. The incubation of [3H]CP with this subcellular fraction led to the binding of a radiolabeled compound to the microsomal lipids, whereas no binding occurred when CP-oxamic acid was incubated with the microsomes. Enzymatic hydrolysis of the microsome lipid fraction with phospholipase D from Streptomyces chromofuscus released CP-oxamylethanolamine.(ABSTRACT TRUNCATED AT 250 WORDS)
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Following subcutaneous administration of [3H]chloramphenicol (CP) to duck, HPLC and TLC analyses showed that the two most important metabolites in 0- to 24-hr excreta were CP-oxamic acid and CP-alcohol, which together accounted for about one-third of the radioactivity therein. The remainder was due to unchanged CP (15% dose), CP-base (5% dose), and various metabolites representing < 4% dose each. Among these, CP-glucuronide and CP-sulfate have been previously isolated in mammals. In addition to these metabolites, several previously unreported in vivo CP biotransformation products were identified in this study by HPLC and MS comparison with synthetic reference compounds. These new metabolites were unequivocally identified as 1-O-monoacetyl CP, CP-1,3-diacetate, N-acetyl CP-base, CP-oxamylglycine, and CP-oxamylethanolamine. Besides these formally identified compounds, the CP-phosphate structure was tentatively assigned to a conjugate metabolite resistant to beta-glucuronidase and sulfatase hydrolysis. The possible origin of these metabolites is discussed extensively.
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