title: Menadione enhances oxyradical formation in earthworm extracts: vulnerability of earthworms to quinone toxicity.
abstract: NAD(P)H-cytochrome c reductase activities have been determined in the earthworms, L. rubellus and A. chlorotica, extracts. Menadione (0.35 mM, maximum concentration tested) was found to stimulate the rates of NADPH- and NADH-dependent cytochrome c reduction by three- and twofold, respectively. Superoxide dismutase (SOD) inhibited completely this menadione-mediated stimulation, suggesting that *O2- is involved in the redox cycling of menadione. However, SOD had no effect on the basal activity (activity in the absence of quinone) in the case of NADH-dependent cytochrome c reduction, whereas it partially inhibited the basal activity of NADPH-cytochrome c reduction. This indicates direct electron transfer in the former case and the formation of superoxide anion in the latter. DT-diaphorase, measured as the dicumarol-inhibitable part of menadione reductase activity, was not detectable in the earthworms' extracts. In contrast, it was found that DT-diaphorase represents about 70% of the menadione reductase activities in the freshwater mussel, Dreissena polymorpha. The results of this work suggest that earthworms, compared with mussels, could be more vulnerable to oxidative stress from quinones due to lack, or very low level of DT-diaphorase, an enzyme considered to play a significant role in the detoxification of quinones. On the contrary, mussels have efficient DT-diaphorase, which catalyzes two-electron reduction of menadione directly to hydroquinone, thus circumventing the formation of semiquinone.
authors.affiliation: Institute for Inland Water management and Waste Water Treatment (RIZA), P.O. Box 17, 8200 AA Lelystad, The Netherlands. a.osman@riza.rws.minvenw.nl
authors.forename: A M
authors.initials: AM
authors.lastname: Osman
authors.forename: P J
authors.initials: PJ
authors.lastname: Den Besten
authors.forename: P C M
authors.initials: PC
authors.lastname: van Noort
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000633:toxicity*; D001728:Dicumarol / Q000494:pharmacology; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D009835:Oligochaeta / Q000201:enzymology* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism; D014020:Tissue Extracts / Q000378:metabolism*; D024483:Vitamin K 3 / Q000494:pharmacology*
chemical_list: D016227:Benzoquinones; D017382:Reactive Oxygen Species; D014020:Tissue Extracts; C004532:quinone; D024483:Vitamin K 3; D001728:Dicumarol; D013482:Superoxide Dismutase; D009251:NADPH-Ferrihemoprotein Reductase; D016660:NAD(P)H Dehydrogenase (Quinone)