title: Chicken fetal liver DNA damage and adduct formation by activation-dependent DNA-reactive carcinogens and related compounds of several structural classes.
abstract: The chicken egg genotoxicity assay (CEGA), which utilizes the liver of an intact and aseptic embryo-fetal test organism, was evaluated using four activation-dependent DNA-reactive carcinogens and four structurally related less potent carcinogens or non-carcinogens. In the assay, three daily doses of test substances were administered to eggs containing 9-11-day-old fetuses and the fetal livers were assessed for two endpoints, DNA breaks using the alkaline single cell gel electrophoresis (comet) assay and DNA adducts using the (32)P-nucleotide postlabeling (NPL) assay. The effects of four carcinogens of different structures requiring distinct pathways of bioactivation, i.e., 2-acetylaminofluorene (AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (B[a]P), and diethylnitrosamine (DEN), were compared with structurally related non-carcinogens fluorene (FLU) and benzo[e]pyrene (B[e]P) or weak carcinogens, aflatoxin B2 (AFB2) and N-nitrosodiethanolamine (NDELA). The four carcinogens all produced DNA breaks at microgram or low milligram total doses, whereas less potent carcinogens and non-carcinogens yielded borderline or negative results, respectively, at higher doses. AAF and B[a]P produced DNA adducts, whereas none was found with the related comparators FLU or B[e]P, consistent with comet results. DEN and NDELA were also negative for adducts, as expected in the case of DEN for an alkylating agent in the standard NPL assay. Also, AFB1 and AFB2 were negative in NPL, as expected, due to the nature of ring opened aflatoxin adducts, which are resistant to enzymatic digestion. Thus, the CEGA, using comet and NPL, is capable of detection of the genotoxicity of diverse DNA-reactive carcinogens, while not yielding false positives for non-carcinogens.
authors: New York Medical College, Chemical Safety Laboratory, Department of Pathology, Valhalla, New York 10595 gary_williams@nymc.edu.
authors: Gary M
authors: GM
authors: Williams
authors: New York Medical College, Chemical Safety Laboratory, Department of Pathology, Valhalla, New York 10595.
authors: Jian-Dong
authors: JD
authors: Duan
authors: New York Medical College, Chemical Safety Laboratory, Department of Pathology, Valhalla, New York 10595.
authors: Klaus D
authors: KD
authors: Brunnemann
authors: New York Medical College, Chemical Safety Laboratory, Department of Pathology, Valhalla, New York 10595.
authors: Michael J
authors: MJ
authors: Iatropoulos
authors: Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorfer Strasse 65, 88379 Biberach an der Riss, Germany.
authors: Esther
authors: E
authors: Vock
authors: Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorfer Strasse 65, 88379 Biberach an der Riss, Germany.
authors: Ulrich
authors: U
authors: Deschl
mesh_terms: D000818:Animals; D002273:Carcinogens / Q000737:chemistry / Q000633:toxicity; D002642:Chick Embryo; D020552:Comet Assay; D018736:DNA Adducts / Q000187:drug effects; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D008099:Liver / Q000187:drug effects* / Q000196:embryology / Q000473:pathology; D059010:Single-Cell Analysis; D013329:Structure-Activity Relationship
chemical_list: D002273:Carcinogens; D018736:DNA Adducts