Monophosphate 32P-postlabeling assay of DNA adducts from 1,2:3,4-diepoxybutane, the most genotoxic metabolite of 1,3-butadiene: in vitro methodological studies and in vivo dosimetry

Sperm quality and DNA integrity of coke oven workers exposed to polycyclic aromatic hydrocarbons

“…Radioactive labeled modified nucleotides were mapped by multidirectional anionexchange thin-layer chromatography (TLC) on polyethyleneimine-cellulose sheets [28]. The 32 P-labeled I-compounds were visualized by screenenhanced autoradiography at -80°C using Kodak BioMax XAR film (Sigma-Aldrich, USA) or with the aid of an InstantImager (Packard Instruments, USA).…”

“…The 32 P-labeled I-compounds were visualized by screenenhanced autoradiography at -80°C using Kodak BioMax XAR film (Sigma-Aldrich, USA) or with the aid of an InstantImager (Packard Instruments, USA). Radioactivity of TLC fractions from individual sperm samples was determined with the aid of an InstantImager electronic autoradiography system as previously described [28]. The extent of covalent DNA adducts was estimated by calculating relative adduct labeling values from the sample count rates, the amount of DNA assayed (expressed as pmol DNA monomer units or DNA-P), and the specific activity of [γ-32 P]ATP according to Zhou et al [29].…”

Mutagenic potential of adenine N6 adducts of monoepoxide and diolepoxide derivatives of butadiene

“…Koc et al [1999] hypothesized that BDO 2 is rapidly hydrolyzed to BDE in the endoplasmic reticulum. An N 6 -trihydroxy adenine derivative was identified by Leuratti et al [1994] in the DNA from CHO cells exposed to BDO 2 and in vivo studies using 32 P-postlabeling methodology showed that when mouse skin was treated with BDO 2 , the major adducts detected comigrated with adenine adducts formed in in vitro studies [Mabon et al, 1996]. Advances in synthetic organic chemistry facilitate the production of site-and stereospecific adducts of these reactive metabolites and allow study of their role in the mutagenic process.…”

Tissue-Specific Attenuation of Endogenous DNA I-Compounds in Rats by Carcinogen Azoxymethane: Possible Role of Dietary Fish Oil in Colon Cancer Prevention

“…After treatment of the mixture with nuclease P1 to convert normal nucleotides to nucleosides, modified nucleotides (Xp) were converted to 5V -32 P-labeled deoxyribonucleoside 3V ,5V -bisphosphates (pXp) by incubation with carrier-free [g-32 P]ATP and polynucleotide kinase. Radioactively labeled modified nucleotides were mapped by multidirectional anion-exchange TLC on polyethyleneiminecellulose sheets (44). Labeled products were purified and partially resolved by one-dimensional development with solvent 1 (Table 2) overnight (D1).…”