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TO establish its physiological role, information is necessary on endogenous MT levels in various tissues. Current methods for the determination of MT are either laborious or relative insensitive. In this communication, we report on a rapid and sensitive assay of MT which is based on the cadmium (Cd) saturation method of Onosaka [Onosaka and Cherian (1982), Tox. Appl. Pharmacol. 63, 270 I. The method takes advantage of the efficient binding of Cd to MT at a definite molar ratio. After separation of MT from excess Cd and Cd-binding ligands other than MT by the addition of hemoglobin and subsequent heating, MT is quantified by the determination of Cd with atomic absorption spectroscopy.
Moreover, when 3H- M- DNM was incubated with primary rat hepatocytes and their DNA was isolated and enzymatically hydrolysed, formation of DNA adducts was observed by HPLC analysis. These data suggest that, in contrast to DNM and ADM, the new anthracyclines, M-DNM, CN-DNM and CN-ADM, can be metabolically activated to generate genotoxic intermediates which react covalently with DNA. most apparent lesions observed were neural tube defects (exencephaly), fetal weight retardation and embryolethality.
Cancer Res. 43:992,1983). , Cancer Res. 42:919,1982). Inhibition of cooxidation with increasing concentrations of indomethacin (100-400~) leads to a decrease of protein bound radioactivity and Z,Z-DIES, but to increased amounts of cis-DES (Z-DES). Time couse studies with native enzyme in the absence of exogenous AA show very efficient and ra- pid isomerization (Z-DES 60% within 3 min); this reaction is inhibited by phenidone, another inhibitor of PGS. Isomerization of E-DES to Z-DES is not accompanied by the formation of protein-bound material and therefore places different toxicological significance on this pathway compared to the cooxidation reaction mediated by the same enzyme.
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