Principles of Genetic Toxicology

1 Origins of Genetic Toxicology.- The History of Genetic Toxicology.- Technology Transfer to Applied Genetics.- The Components of Genetic Toxicology.- References.- 2 Fundamentals of Genetic Toxicology.- Basic Genetics for Toxicologists.- Gene Structure.- Gene Function.- The Cell Cycle and Chromosome Mechanics in Somatic and Germ Cells.- DNA Alterations Resulting in Genotoxic Effects in Cells: Mechanisms and Categorization.- A Classification Scheme for Genotoxic Effects.- Repair of DNA Damage.- References.- 3 The Consequences of Genotoxic Effects in Humans and Other Mammals.- Gene Pool Consequences.- The Direct Effect of Mutagens on the Transmissible Gene Pool.- The Relationship of Genotoxic Effects to Other Toxicologic End Points.- Relationship of Potency between Mutagenicity Assays and in Vivo Oncogenicity.- References.- 4 Screening Chemicals for Genotoxic Properties.- and Background.- Characteristics of Adequate Screening Tests.- The Type(s) and Number(s) of End Points Detected.- The Metabolic Capability of the Test or Associated Activating System.- Reliability/Reproducibility.- Facilities.- Strategies for Test Battery Development.- General Philosophy.- Approaches to Test Battery Development.- Recommendations for Group I Tests.- Interpretation of Data from Screening Tests.- The Use of Controls in in Vitro and in Vivo Testing.- Approaches to Data Analysis and Interpretation.- References.- 5 Genetic Risk Estimation.- Definition of Risk.- Risk Estimation in Somatic and Germ Cells.- Somatic Cell Risk.- Germ Cell Risk.- Relationship of the Route of Exposure to Total Body Dose.- Relationship of the Route of Exposure to Metabolism.- Blood/Gonadal Barriers.- Molecular Dosimetry of Chemical Mutagens.- Measuring the Genotoxic Effect.- Approaches to Germ Cell Risk Using Animal Models.- Extrapolating Data from Short-Term Tests to Humans.- Extrapolation of in Vitro Data Directly to in Vivo Responses.- Threshold.- References.- 6 Applications of Genetic Toxicology to Environmental and Human Monitoring.- Monitoring Techniques.- Occupational Monitoring.- Environmental Monitoring.- References.- 7 The Genetic Toxicology Laboratory.- General Laboratory Areas.- Chemical Storage and Waste Disposal.- Laboratory Safety and Employee Monitoring.- Good Laboratory Practices Requirements.- References.- 8 Descriptions of Genetic Toxicology Assays.- General Classification of Genetic Toxicology Assays.- Description of Common Assays for Gene Mutation.- Microbial.- Mammalian Cells in Vitro.- Insects.- Mammals.- Tests for Chromosome Aberrations.- Microbial.- Mammalian Cells in Vitro.- Insect Tests for Chromosome Effects.- Mammals.- Tests for Primary DNA Damage.- Microbial Tests.- Cultured Mammalian Cell Repair Assays.- SCE Analysis.- Insects.- In Vivo Mammalian Primary DNA Damage.- In Vitro Cell Transformation.- References.- 9 Sample Study Designs.- Selected Study Designs.- Protocol I: Ames Salmonella/Microsome Plate Assay.- Materials.- Test Procedures.- Evaluation Criteria.- Surviving Populations.- Dose-Response Phenomena.- Control Tests.- Evaluation Criteria for Ames Assay.- Relation between Mutagenicity and Carcinogenicity.- References.- Protocol 2: Mitotic Recombination in S. cerevisiae Strain D3.- Dosing Procedure.- Method.- Evaluation Criteria.- References.- Protocol 3: Mitotic Gene Conversion in S. cerevisiae Strain D4.- Dosing Procedure.- Method.- General Evaluation Criteria.- Surviving Populations.- Dose-Response Phenomena.- Control Tests.- Evaluation Criteria for the Preincubation Plate Incorporation Assays Using Yeast Strain D4.- Reproducibility.- References.- Protocol 4: Mitotic Recombination in S. cerevisiae Strain D5.- Dosing Procedure.- Method.- Evaluation Criteria.- References.- Protocol 5: L5178Y TK+/?Mouse Lymphoma Forward Mutation Assay.- Materials.- Indicator Cells.- Media.- Control Articles.- Sample Forms.- Experimental Design.- Dose Selection.- Mutagenicity Testing.- Preparation of 9000g Supernatant (S9).- Assay Acceptance Criteria.- Assay Evaluation Criteria.- Reference.- Protocol 6: Chromosome Aberrations in Chinese Hamster Ovary Cells.- Materials.- Indicator Cells.- Medium.- Control Articles.- Experimental Design.- Toxicity and Dose Determination.- Cell Treatment.- Preparation of S9 Reaction Mixture.- Staining and Scoring of Slides.- Evaluation Criteria.- Reference.- Protocol 7: Sister Chromatid Exchange in Human Lymphocytes.- Materials.- Experimental Design.- Option 1.- Option 2.- Staining and Scoring of Slides.- Evaluation Criteria.- Reference.- Protocol 8: Sister Chromatid Exchange in Chinese Hamster Ovary Cells.- Materials.- Experimental Design.- Toxicity and Dose Determination.- Cell Treatment.- Preparation of S9 Reaction Mixture.- Staining and Scoring of Slides.- Evaluation Criteria.- References.- Protocol 9: Determination of Unscheduled DNA Synthesis in Human WI-38 Cells.- Materials.- Experimental Design.- Dose Selection.- Nonactivation Assay.- Activation Assay.- Extraction of DNA.- DNA Analysis.- Preparation of 9000g Supernatant (S9).- Evaluation Criteria.- Reference.- Protocol 10: Unscheduled DNA Synthesis in Rat Liver Primary Cell Cultures.- Materials.- Experimental Design.- Dose Selection.- UDS Assay Method.- Evaluation Criteria.- Reference.- Protocol 11: In Vitro Transformation of BALB/3T3 Cells.- Materials.- Experimental Design.- Dose Selection.- Transformation Assay.- Scoring of Transformed Foci.- Assay Acceptance Criteria.- Assay Evaluation Criteria.- References.- Protocol 12: Microbial Host-Mediated Assay.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Route of Administration.- Test and Control Articles.- Bacteria Administration and Recovery.- Cell Plating for Survival and Mutant Determinations.- References.- Protocol 13: Bone Marrow Cytogenetic Analysis in Rats.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Route of Administration.- Methodology.- Evaluation Criteria.- References.- Protocol 14: Dominant Lethal Assay in Rats.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Route of Administration.- Methodology.- References.- Protocol 15: Heritable Translocation Assay in Mice.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Methodology.- References.- Protocol 16: Mouse Micronucleus Assay.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Dosing Schedule and Route of Administration.- Extraction of Bone Marrow.- Preparation of Slides.- Screening the Slides.- Evaluation Criteria.- References.- Protocol 17: Mouse Sperm head Abnormalities.- Materials.- Experimental Design.- Animal Husbandry.- Dose Selection.- Dosing Schedule and Route of Administration.- Preparation of Sperm Suspension.- Evaluation Criteria.- References.- Protocol 18: Somatic Mutation Assay in the Mouse (Spot Test).- Materials.- Animals.- Control Articles.- Experimental Design.- Animal Husbandry.- Dose Selection.- Route of Administration.- Treatment.- Spot Observation.- References.- Protocol 19: Test for Loss of X and Y Chromosomes in D. melanogaster.- Materials.- Experimental Design.- Solubility Testing.- Palatability and Toxicity Testing.- Fertility Testing.- Test Size.- Procedure.- Collection of Males.- Dosing.- Mating and Scoring.- Data Analysis.- Reference.- Protocol 20: Heritable Translocation Test in D. melanogaster.- Materials.- Experimental Design.- Solubility Testing.- Palatability and Toxicity Testing.- Fertility Testing.- Test Size.- Procedure.- Collection of Males.- Dosing.- Mating and Scoring.- Data Analysis.- Reference.- Protocol 21: Sex-Linked Recessive Lethal Test in D. melanogaster.- Materials.- Experimental Design.- Solubility Testing.- Palatability and Toxicity Testing.- Fertility Testing.- Test Size.- Procedure.- Collection of Males.- Dosing.- Mating and Scoring.- Data Analysis.- References.- Appendices.- Appendix A: Preparation of S9 Liver Homogenates.- Induction of Microsomal Enzymes.- Preparation of the S9 Fraction.- Preparation of the S9 Mix.- References.- References.- Appendix C: Selected References and Reviews of Genetics and Genetic Toxicology.