Kooperativer Bibliotheksverbund

UID:

Format: xxii, 434 Seiten , Illustrationen, Diagramme

ISBN: 0128007648 , 9780128007648

Note: Literaturangaben

Language: English

Subjects: Chemistry/Pharmacy , Biology

Keywords: Gentoxikologie ; Testen ; Labortechnik

UID:

Format: 1 online resource (458 pages) : , illustrations, tables

ISBN: 0-12-801006-1

Note: Front Cover -- Genetic Toxicology Testing -- Copyright Page -- Contents -- List of Contributors -- Foreword -- Preface -- References -- 1 A Practical Guide to Genetic Toxicology Testing -- 1.1 Introduction -- References -- 2 General Recommendations -- 2.1 Establishing a New Assay -- 2.1.1 Method Set-Up -- 2.1.2 Method Validation -- 2.2 Spreadsheets and Manipulation of Results -- 2.3 Laboratory Historical Control Databases -- 2.3.1 Vehicle/Negative Control Database -- 2.3.2 Positive Control Database -- 2.3.3 Spreadsheet Calculations -- 2.4 Use of Computer Systems -- 2.5 Study Design -- 2.6 Evaluation Criteria -- 2.6.1 Valid Assay -- 2.6.2 Criteria for Interpretation of Results -- 2.6.3 Statistical Analysis -- 2.7 Organization of SOPs -- 2.8 Planning a Study -- 2.9 Preparing a Protocol Complying with GLP -- 2.9.1 Standardized Boilerplate Protocols -- 2.10 Collecting Results -- 2.10.1 Data Tabulation -- 2.10.2 Presentation of Report Tables -- 2.11 Reports -- 2.12 Training -- 2.13 Improving Quality and Efficiency -- 2.13.1 Improving Quality -- 2.13.2 Minimizing the Potential Problems on Studies -- 2.13.3 Reducing the Need for Repetition of Parts of the Study -- 2.13.4 Accommodating Repeat or Supplementary Tests -- 2.13.5 Running More Studies in a Given Timeframe -- 2.13.6 Timeframe for Routine Studies -- 2.13.7 Reducing the Effort Needed to Prepare Protocols, Tables, and Reports -- 2.13.8 Reducing the Effort Needed to Perform the Study -- 2.13.9 Reducing Costs or Labor Requirement -- 2.14 QA -- 2.15 Qualifying a Contract Laboratory -- 2.16 Responsibilities of the Study Monitor -- References -- 3 Formulation of Test Articles -- 3.1 Introduction -- 3.1.1 Safety -- 3.1.2 Selecting an Appropriate Formulation -- 3.2 Formulation Laboratories -- 3.2.1 Designing and Equipping a Genetic Toxicology Formulation Area -- 3.2.1.1 Hoods -- 3.2.1.2 Storage equipment. , 3.2.2 Personal Protective Clothing -- 3.2.2.1 Standard -- 3.2.2.2 Capital equipment -- 3.2.2.3 Consumables -- 3.3 Safety Data Sheets -- 3.4 Receipt of the Test Article -- 3.5 Formulation Types and Planning -- 3.6 Solubility and In Vitro Compatibility Testing -- 3.6.1 Introduction -- 3.6.2 Choice of Solvent -- 3.6.3 Solubility Testing -- 3.6.4 Calculations and Checking: Small-Volume (In Vitro) Assays -- 3.6.5 Compatibility of Formulation with Culture Medium -- 3.7 Formulation of Dose Solutions -- 3.8 Formulation of Bulk Formulations -- 3.9 Formulation of Suspensions -- 3.9.1 Aqueous Suspending Agents -- 3.9.2 Large Volume Suspensions -- 3.9.3 Small Volume Suspensions -- 3.10 Chemical Analysis and Stability -- References -- 4 The Bacterial Reverse Mutation Test -- 4.1 Introduction -- 4.2 History -- 4.3 Fundamentals -- 4.4 Equipment -- 4.5 Consumables -- 4.6 Reagents and Recipes -- 4.6.1 Ampicillin 2 µg/disc -- 4.6.2 Biotin 0.37 mg/mL -- 4.6.3 Crystal Violet 5 µg/disc -- 4.6.4 Glucose 0.4 g/mL -- 4.6.5 G6P 1M: Glucose-6-Phosphate -- 4.6.6 HBT: 500 µM Histidine, 500µM Biotin, 500 µM Tryptophan Solution -- 4.6.7 Histidine HCl.H2O 5 mg/mL -- 4.6.8 KMg -- 4.6.9 MGA Plates -- 4.6.10 Minimal Glucose Master (MGM, MGMA and MGMAT) Plates -- 4.6.11 NADP 0.1 M -- 4.6.12 Nutrient Agar Plates -- 4.6.13 Nutrient Broth -- 4.6.14 Phosphate Buffer 0.2 M pH 7.4 -- 4.6.15 Positive Control and Diagnostic Mutagen Solutions -- 4.6.16 S9 Fraction -- 4.6.17 S9 Mix -- 4.6.18 Tetracycline 1 µg/disc -- 4.6.19 Top Agar Incomplete: TAI -- 4.6.20 Top Agar Complete: TAC -- 4.6.21 Tryptophan 5 mg/mL -- 4.6.22 VB Salts 50×: Vogel-Bonner Salts -- 4.7 Suggested Phases in Development of the Test -- 4.8 The Bacterial Strains -- 4.8.1 Genotypes of Routinely Used Strains -- 4.8.2 Obtaining the Tester Strains -- 4.8.3 Receipt of Bacterial Strains -- 4.8.4 Phenotyping of New Isolates. , 4.8.5 Freezing of Selected Isolates -- 4.8.6 Diagnostic Mutagen Test -- 4.9 Routine Testing -- 4.9.1 Designing a Study -- 4.9.1.1 Metabolic activation system -- 4.9.2 Test Article Considerations -- 4.9.2.1 Solvent selection -- 4.9.2.2 Dose volumes -- 4.9.2.3 Dose levels -- 4.9.3 Positive Controls -- 4.10 Standard Test Procedures -- 4.10.1 Plate Incorporation Method -- 4.10.2 Preincubation Method -- 4.10.3 Standard Study Design -- 4.10.4 Examination of the Plates -- 4.10.5 Interpretation of Results -- 4.10.5.1 Evaluation of toxicity -- 4.10.5.2 Validity of the study -- 4.10.5.3 Criteria for negative/positive/equivocal outcome -- 4.10.5.4 Unexpected and borderline results -- 4.10.6 Presentation of Results -- 4.10.7 Testing of Volatile and Gaseous Compounds -- 4.11 Screening Tests -- 4.11.1 Simplified Test Systems -- 4.11.2 Screening Tests Using Standard Tester Strains -- 4.11.3 Reduced Format Tests Using Standard Tester Strains -- 4.12 Appendix 1: Growing and Monitoring Suspension Cultures -- References -- 5 The Mouse Lymphoma TK Assay -- 5.1 Introduction -- 5.2 History -- 5.3 Provenance of the Cells -- 5.4 Spontaneous Mutation Frequency -- 5.5 Materials -- 5.5.1 Safety -- 5.5.2 Growth Medium -- 5.5.3 Cell Culture -- 5.5.4 Metabolic Activation -- 5.5.5 Test Item -- 5.5.6 Vehicle -- 5.5.7 Positive Controls -- 5.6 Study Design -- 5.6.1 General Test Conditions -- 5.6.2 Preliminary Toxicity Test -- 5.6.3 Main Mutation Test -- 5.6.3.1 Posttreatment procedures -- 5.6.3.2 Expression period -- 5.6.3.2.1 Expression period day 1 -- 5.6.3.2.2 Expression period day 2 -- 5.6.3.3 Viability assessment and mutant selection (microtiter version) -- 5.6.3.3.1 Colony counting (microtiter version) -- 5.6.3.4 Viability assessment and mutant selection (agar version) -- 5.6.3.4.1 Colony counting (agar version) -- 5.6.3.5 Analysis of results. , 5.6.3.5.1 Relative suspension growth -- 5.6.3.5.2 Toxicity assessment -- 5.6.3.5.3 MF assessment (microtiter version) -- 5.6.3.5.4 MF assessment (agar version) -- 5.6.3.6 Acceptance criteria -- 5.7 Evaluation Criteria -- 5.8 Predictivity of the MLA -- References -- 6 The In Vitro Micronucleus Assay -- 6.1 Introduction -- 6.2 Practical Considerations -- 6.2.1 Regulatory Guidelines -- 6.2.2 Good Laboratory Practice (GLP) -- 6.2.3 Cell Types -- 6.2.4 Laboratory Proficiency -- 6.2.5 Controls -- 6.2.6 Metabolic Activation -- 6.2.7 S9 Rat Liver Homogenate -- 6.2.8 Experimental Design -- 6.2.9 Cytotoxicity Measures -- 6.2.9.1 Methods used to determine cytotoxicity in the absence of cytochalasin B [16] (reproduced with permission of the author) -- 6.2.9.2 Method used to determine cytotoxicity in the presence of cytochalasin B -- 6.2.10 Historical Controls -- 6.3 Methods -- 6.3.1 Mononuclear Assay -- 6.3.2 Binuclear Assay -- 6.3.3 Centromeric Labeling -- 6.3.4 Nondisjunction Assay -- 6.4 Materials -- 6.4.1 Mononuclear Assay -- 6.4.2 Binuclear Assay -- 6.4.3 Centromeric Labeling -- 6.4.4 Nondisjunction Assay -- 6.5 Protocols -- 6.5.1 S9 Mix -- 6.5.2 Mononuclear Assay -- 6.5.2.1 Treatment schedules -- 6.5.2.2 Cell culture and treatment -- 6.5.2.3 Slide preparation -- 6.5.2.4 Coding of slides -- 6.5.2.5 Analysis of slides (microscope) -- 6.5.2.6 Analysis of slides (semiautomated scoring) -- Criteria for evaluation -- 6.5.3 Binuclear Assay -- 6.5.3.1 Treatment schedules -- 6.5.3.2 Human peripheral blood lymphocytes -- 6.5.3.3 Donors -- 6.5.3.4 Lymphocyte culture -- 6.5.3.5 Staining and analysis -- 6.5.3.6 Coding of slides -- 6.5.3.7 Analysis of slides -- 6.5.3.8 Evaluation of results (acceptance criteria and statistics) -- 6.5.3.9 Criteria for a valid assay -- 6.5.3.10 Evaluation of data -- 6.5.4 Centromeric Labeling. , 6.5.4.1 FISH using a programmable hotplate such as HYBrite™ or Thermobrite™ -- 6.5.4.2 Alterative protocol for FISH -- 6.5.4.3 Slide checking -- 6.5.4.4 Slide scoring -- 6.5.5 Nondisjunction Assay -- 6.5.5.1 FISH method -- 6.5.5.2 Slide checking -- 6.5.5.3 Slide scoring -- 6.6 Flow Cytometric Method -- 6.6.1 Equipment -- 6.6.2 Consumables -- 6.6.3 Reagents and recipes -- 6.6.4 Suspension Cell Protocol -- 6.6.5 Attachment Cell Protocol -- 6.6.6 Flow Cytometric Data Acquisition -- 6.6.7 Flow Cytometric Data Analysis -- 6.6.7.1 Independent cytotoxicity assessment -- 6.6.7.2 Criteria for a valid assay -- 6.6.7.3 Criteria for positive/negative outcomes -- 6.6.8 Creating an Analysis Template -- 6.6.9 Example Plate Layout -- 6.6.10 Example Results Table -- 6.6.11 Advice for Test Article Exposure -- 6.6.11.1 Suspension cells -- 6.6.11.2 Attachment cells -- 6.6.11.3 Metabolic activation -- 6.6.11.4 Positive controls -- 6.6.12 Use of Multichannel Aspirator with Bridge -- 6.6.13 Plate Placement During Nucleic Acid Dye B Photoactivation -- 6.6.14 Updates and Future Work -- References -- 7 The In Vitro Chromosome Aberration Test -- 7.1 Introduction -- 7.2 History -- 7.3 Fundamentals -- 7.4 Equipment -- 7.5 Consumables and Reagents -- 7.6 Reagents and Recipes -- 7.6.1 Colcemid 10 µg/mL in PBS -- 7.6.2 Fix -- 7.6.3 F-12 Complete -- 7.6.4 Freezing Medium 10% (CHO Cells) -- 7.6.5 Hypotonic Solution (0.075 M KCl) -- 7.6.6 Heparin Sodium 1000 U/mL -- 7.6.7 G6P 1 M: Glucose-6-Phosphate -- 7.6.8 KMg -- 7.6.9 NADP 0.1 M -- 7.6.10 PHA M Form (Phytohemagglutinin) -- 7.6.11 Phosphate Buffer 0.2 M, pH 7.4 -- 7.6.12 Positive Control Solutions -- 7.6.13 RPMI Complete -- 7.6.14 S9 Fraction -- 7.6.15 S9 Mix -- 7.7 Phases in Development of the Test -- 7.8 Cell Characterization -- 7.8.1 Modal Chromosome Number -- 7.8.2 Mycoplasma -- 7.8.3 Cell-Cycle Time -- 7.9 Routine Testing. , 7.9.1 General Considerations.

Additional Edition: ISBN 0-12-800764-8

Language: English

UID:

Format: 1 online resource (458 pages) : , illustrations, tables

ISBN: 0-12-801006-1

Note: Front Cover -- Genetic Toxicology Testing -- Copyright Page -- Contents -- List of Contributors -- Foreword -- Preface -- References -- 1 A Practical Guide to Genetic Toxicology Testing -- 1.1 Introduction -- References -- 2 General Recommendations -- 2.1 Establishing a New Assay -- 2.1.1 Method Set-Up -- 2.1.2 Method Validation -- 2.2 Spreadsheets and Manipulation of Results -- 2.3 Laboratory Historical Control Databases -- 2.3.1 Vehicle/Negative Control Database -- 2.3.2 Positive Control Database -- 2.3.3 Spreadsheet Calculations -- 2.4 Use of Computer Systems -- 2.5 Study Design -- 2.6 Evaluation Criteria -- 2.6.1 Valid Assay -- 2.6.2 Criteria for Interpretation of Results -- 2.6.3 Statistical Analysis -- 2.7 Organization of SOPs -- 2.8 Planning a Study -- 2.9 Preparing a Protocol Complying with GLP -- 2.9.1 Standardized Boilerplate Protocols -- 2.10 Collecting Results -- 2.10.1 Data Tabulation -- 2.10.2 Presentation of Report Tables -- 2.11 Reports -- 2.12 Training -- 2.13 Improving Quality and Efficiency -- 2.13.1 Improving Quality -- 2.13.2 Minimizing the Potential Problems on Studies -- 2.13.3 Reducing the Need for Repetition of Parts of the Study -- 2.13.4 Accommodating Repeat or Supplementary Tests -- 2.13.5 Running More Studies in a Given Timeframe -- 2.13.6 Timeframe for Routine Studies -- 2.13.7 Reducing the Effort Needed to Prepare Protocols, Tables, and Reports -- 2.13.8 Reducing the Effort Needed to Perform the Study -- 2.13.9 Reducing Costs or Labor Requirement -- 2.14 QA -- 2.15 Qualifying a Contract Laboratory -- 2.16 Responsibilities of the Study Monitor -- References -- 3 Formulation of Test Articles -- 3.1 Introduction -- 3.1.1 Safety -- 3.1.2 Selecting an Appropriate Formulation -- 3.2 Formulation Laboratories -- 3.2.1 Designing and Equipping a Genetic Toxicology Formulation Area -- 3.2.1.1 Hoods -- 3.2.1.2 Storage equipment. , 3.2.2 Personal Protective Clothing -- 3.2.2.1 Standard -- 3.2.2.2 Capital equipment -- 3.2.2.3 Consumables -- 3.3 Safety Data Sheets -- 3.4 Receipt of the Test Article -- 3.5 Formulation Types and Planning -- 3.6 Solubility and In Vitro Compatibility Testing -- 3.6.1 Introduction -- 3.6.2 Choice of Solvent -- 3.6.3 Solubility Testing -- 3.6.4 Calculations and Checking: Small-Volume (In Vitro) Assays -- 3.6.5 Compatibility of Formulation with Culture Medium -- 3.7 Formulation of Dose Solutions -- 3.8 Formulation of Bulk Formulations -- 3.9 Formulation of Suspensions -- 3.9.1 Aqueous Suspending Agents -- 3.9.2 Large Volume Suspensions -- 3.9.3 Small Volume Suspensions -- 3.10 Chemical Analysis and Stability -- References -- 4 The Bacterial Reverse Mutation Test -- 4.1 Introduction -- 4.2 History -- 4.3 Fundamentals -- 4.4 Equipment -- 4.5 Consumables -- 4.6 Reagents and Recipes -- 4.6.1 Ampicillin 2 µg/disc -- 4.6.2 Biotin 0.37 mg/mL -- 4.6.3 Crystal Violet 5 µg/disc -- 4.6.4 Glucose 0.4 g/mL -- 4.6.5 G6P 1M: Glucose-6-Phosphate -- 4.6.6 HBT: 500 µM Histidine, 500µM Biotin, 500 µM Tryptophan Solution -- 4.6.7 Histidine HCl.H2O 5 mg/mL -- 4.6.8 KMg -- 4.6.9 MGA Plates -- 4.6.10 Minimal Glucose Master (MGM, MGMA and MGMAT) Plates -- 4.6.11 NADP 0.1 M -- 4.6.12 Nutrient Agar Plates -- 4.6.13 Nutrient Broth -- 4.6.14 Phosphate Buffer 0.2 M pH 7.4 -- 4.6.15 Positive Control and Diagnostic Mutagen Solutions -- 4.6.16 S9 Fraction -- 4.6.17 S9 Mix -- 4.6.18 Tetracycline 1 µg/disc -- 4.6.19 Top Agar Incomplete: TAI -- 4.6.20 Top Agar Complete: TAC -- 4.6.21 Tryptophan 5 mg/mL -- 4.6.22 VB Salts 50×: Vogel-Bonner Salts -- 4.7 Suggested Phases in Development of the Test -- 4.8 The Bacterial Strains -- 4.8.1 Genotypes of Routinely Used Strains -- 4.8.2 Obtaining the Tester Strains -- 4.8.3 Receipt of Bacterial Strains -- 4.8.4 Phenotyping of New Isolates. , 4.8.5 Freezing of Selected Isolates -- 4.8.6 Diagnostic Mutagen Test -- 4.9 Routine Testing -- 4.9.1 Designing a Study -- 4.9.1.1 Metabolic activation system -- 4.9.2 Test Article Considerations -- 4.9.2.1 Solvent selection -- 4.9.2.2 Dose volumes -- 4.9.2.3 Dose levels -- 4.9.3 Positive Controls -- 4.10 Standard Test Procedures -- 4.10.1 Plate Incorporation Method -- 4.10.2 Preincubation Method -- 4.10.3 Standard Study Design -- 4.10.4 Examination of the Plates -- 4.10.5 Interpretation of Results -- 4.10.5.1 Evaluation of toxicity -- 4.10.5.2 Validity of the study -- 4.10.5.3 Criteria for negative/positive/equivocal outcome -- 4.10.5.4 Unexpected and borderline results -- 4.10.6 Presentation of Results -- 4.10.7 Testing of Volatile and Gaseous Compounds -- 4.11 Screening Tests -- 4.11.1 Simplified Test Systems -- 4.11.2 Screening Tests Using Standard Tester Strains -- 4.11.3 Reduced Format Tests Using Standard Tester Strains -- 4.12 Appendix 1: Growing and Monitoring Suspension Cultures -- References -- 5 The Mouse Lymphoma TK Assay -- 5.1 Introduction -- 5.2 History -- 5.3 Provenance of the Cells -- 5.4 Spontaneous Mutation Frequency -- 5.5 Materials -- 5.5.1 Safety -- 5.5.2 Growth Medium -- 5.5.3 Cell Culture -- 5.5.4 Metabolic Activation -- 5.5.5 Test Item -- 5.5.6 Vehicle -- 5.5.7 Positive Controls -- 5.6 Study Design -- 5.6.1 General Test Conditions -- 5.6.2 Preliminary Toxicity Test -- 5.6.3 Main Mutation Test -- 5.6.3.1 Posttreatment procedures -- 5.6.3.2 Expression period -- 5.6.3.2.1 Expression period day 1 -- 5.6.3.2.2 Expression period day 2 -- 5.6.3.3 Viability assessment and mutant selection (microtiter version) -- 5.6.3.3.1 Colony counting (microtiter version) -- 5.6.3.4 Viability assessment and mutant selection (agar version) -- 5.6.3.4.1 Colony counting (agar version) -- 5.6.3.5 Analysis of results. , 5.6.3.5.1 Relative suspension growth -- 5.6.3.5.2 Toxicity assessment -- 5.6.3.5.3 MF assessment (microtiter version) -- 5.6.3.5.4 MF assessment (agar version) -- 5.6.3.6 Acceptance criteria -- 5.7 Evaluation Criteria -- 5.8 Predictivity of the MLA -- References -- 6 The In Vitro Micronucleus Assay -- 6.1 Introduction -- 6.2 Practical Considerations -- 6.2.1 Regulatory Guidelines -- 6.2.2 Good Laboratory Practice (GLP) -- 6.2.3 Cell Types -- 6.2.4 Laboratory Proficiency -- 6.2.5 Controls -- 6.2.6 Metabolic Activation -- 6.2.7 S9 Rat Liver Homogenate -- 6.2.8 Experimental Design -- 6.2.9 Cytotoxicity Measures -- 6.2.9.1 Methods used to determine cytotoxicity in the absence of cytochalasin B [16] (reproduced with permission of the author) -- 6.2.9.2 Method used to determine cytotoxicity in the presence of cytochalasin B -- 6.2.10 Historical Controls -- 6.3 Methods -- 6.3.1 Mononuclear Assay -- 6.3.2 Binuclear Assay -- 6.3.3 Centromeric Labeling -- 6.3.4 Nondisjunction Assay -- 6.4 Materials -- 6.4.1 Mononuclear Assay -- 6.4.2 Binuclear Assay -- 6.4.3 Centromeric Labeling -- 6.4.4 Nondisjunction Assay -- 6.5 Protocols -- 6.5.1 S9 Mix -- 6.5.2 Mononuclear Assay -- 6.5.2.1 Treatment schedules -- 6.5.2.2 Cell culture and treatment -- 6.5.2.3 Slide preparation -- 6.5.2.4 Coding of slides -- 6.5.2.5 Analysis of slides (microscope) -- 6.5.2.6 Analysis of slides (semiautomated scoring) -- Criteria for evaluation -- 6.5.3 Binuclear Assay -- 6.5.3.1 Treatment schedules -- 6.5.3.2 Human peripheral blood lymphocytes -- 6.5.3.3 Donors -- 6.5.3.4 Lymphocyte culture -- 6.5.3.5 Staining and analysis -- 6.5.3.6 Coding of slides -- 6.5.3.7 Analysis of slides -- 6.5.3.8 Evaluation of results (acceptance criteria and statistics) -- 6.5.3.9 Criteria for a valid assay -- 6.5.3.10 Evaluation of data -- 6.5.4 Centromeric Labeling. , 6.5.4.1 FISH using a programmable hotplate such as HYBrite™ or Thermobrite™ -- 6.5.4.2 Alterative protocol for FISH -- 6.5.4.3 Slide checking -- 6.5.4.4 Slide scoring -- 6.5.5 Nondisjunction Assay -- 6.5.5.1 FISH method -- 6.5.5.2 Slide checking -- 6.5.5.3 Slide scoring -- 6.6 Flow Cytometric Method -- 6.6.1 Equipment -- 6.6.2 Consumables -- 6.6.3 Reagents and recipes -- 6.6.4 Suspension Cell Protocol -- 6.6.5 Attachment Cell Protocol -- 6.6.6 Flow Cytometric Data Acquisition -- 6.6.7 Flow Cytometric Data Analysis -- 6.6.7.1 Independent cytotoxicity assessment -- 6.6.7.2 Criteria for a valid assay -- 6.6.7.3 Criteria for positive/negative outcomes -- 6.6.8 Creating an Analysis Template -- 6.6.9 Example Plate Layout -- 6.6.10 Example Results Table -- 6.6.11 Advice for Test Article Exposure -- 6.6.11.1 Suspension cells -- 6.6.11.2 Attachment cells -- 6.6.11.3 Metabolic activation -- 6.6.11.4 Positive controls -- 6.6.12 Use of Multichannel Aspirator with Bridge -- 6.6.13 Plate Placement During Nucleic Acid Dye B Photoactivation -- 6.6.14 Updates and Future Work -- References -- 7 The In Vitro Chromosome Aberration Test -- 7.1 Introduction -- 7.2 History -- 7.3 Fundamentals -- 7.4 Equipment -- 7.5 Consumables and Reagents -- 7.6 Reagents and Recipes -- 7.6.1 Colcemid 10 µg/mL in PBS -- 7.6.2 Fix -- 7.6.3 F-12 Complete -- 7.6.4 Freezing Medium 10% (CHO Cells) -- 7.6.5 Hypotonic Solution (0.075 M KCl) -- 7.6.6 Heparin Sodium 1000 U/mL -- 7.6.7 G6P 1 M: Glucose-6-Phosphate -- 7.6.8 KMg -- 7.6.9 NADP 0.1 M -- 7.6.10 PHA M Form (Phytohemagglutinin) -- 7.6.11 Phosphate Buffer 0.2 M, pH 7.4 -- 7.6.12 Positive Control Solutions -- 7.6.13 RPMI Complete -- 7.6.14 S9 Fraction -- 7.6.15 S9 Mix -- 7.7 Phases in Development of the Test -- 7.8 Cell Characterization -- 7.8.1 Modal Chromosome Number -- 7.8.2 Mycoplasma -- 7.8.3 Cell-Cycle Time -- 7.9 Routine Testing. , 7.9.1 General Considerations.

Additional Edition: ISBN 0-12-800764-8

Language: English

UID:

Format: 1 online resource (458 pages) : , illustrations, tables

ISBN: 0-12-801006-1

Note: Front Cover -- Genetic Toxicology Testing -- Copyright Page -- Contents -- List of Contributors -- Foreword -- Preface -- References -- 1 A Practical Guide to Genetic Toxicology Testing -- 1.1 Introduction -- References -- 2 General Recommendations -- 2.1 Establishing a New Assay -- 2.1.1 Method Set-Up -- 2.1.2 Method Validation -- 2.2 Spreadsheets and Manipulation of Results -- 2.3 Laboratory Historical Control Databases -- 2.3.1 Vehicle/Negative Control Database -- 2.3.2 Positive Control Database -- 2.3.3 Spreadsheet Calculations -- 2.4 Use of Computer Systems -- 2.5 Study Design -- 2.6 Evaluation Criteria -- 2.6.1 Valid Assay -- 2.6.2 Criteria for Interpretation of Results -- 2.6.3 Statistical Analysis -- 2.7 Organization of SOPs -- 2.8 Planning a Study -- 2.9 Preparing a Protocol Complying with GLP -- 2.9.1 Standardized Boilerplate Protocols -- 2.10 Collecting Results -- 2.10.1 Data Tabulation -- 2.10.2 Presentation of Report Tables -- 2.11 Reports -- 2.12 Training -- 2.13 Improving Quality and Efficiency -- 2.13.1 Improving Quality -- 2.13.2 Minimizing the Potential Problems on Studies -- 2.13.3 Reducing the Need for Repetition of Parts of the Study -- 2.13.4 Accommodating Repeat or Supplementary Tests -- 2.13.5 Running More Studies in a Given Timeframe -- 2.13.6 Timeframe for Routine Studies -- 2.13.7 Reducing the Effort Needed to Prepare Protocols, Tables, and Reports -- 2.13.8 Reducing the Effort Needed to Perform the Study -- 2.13.9 Reducing Costs or Labor Requirement -- 2.14 QA -- 2.15 Qualifying a Contract Laboratory -- 2.16 Responsibilities of the Study Monitor -- References -- 3 Formulation of Test Articles -- 3.1 Introduction -- 3.1.1 Safety -- 3.1.2 Selecting an Appropriate Formulation -- 3.2 Formulation Laboratories -- 3.2.1 Designing and Equipping a Genetic Toxicology Formulation Area -- 3.2.1.1 Hoods -- 3.2.1.2 Storage equipment. , 3.2.2 Personal Protective Clothing -- 3.2.2.1 Standard -- 3.2.2.2 Capital equipment -- 3.2.2.3 Consumables -- 3.3 Safety Data Sheets -- 3.4 Receipt of the Test Article -- 3.5 Formulation Types and Planning -- 3.6 Solubility and In Vitro Compatibility Testing -- 3.6.1 Introduction -- 3.6.2 Choice of Solvent -- 3.6.3 Solubility Testing -- 3.6.4 Calculations and Checking: Small-Volume (In Vitro) Assays -- 3.6.5 Compatibility of Formulation with Culture Medium -- 3.7 Formulation of Dose Solutions -- 3.8 Formulation of Bulk Formulations -- 3.9 Formulation of Suspensions -- 3.9.1 Aqueous Suspending Agents -- 3.9.2 Large Volume Suspensions -- 3.9.3 Small Volume Suspensions -- 3.10 Chemical Analysis and Stability -- References -- 4 The Bacterial Reverse Mutation Test -- 4.1 Introduction -- 4.2 History -- 4.3 Fundamentals -- 4.4 Equipment -- 4.5 Consumables -- 4.6 Reagents and Recipes -- 4.6.1 Ampicillin 2 µg/disc -- 4.6.2 Biotin 0.37 mg/mL -- 4.6.3 Crystal Violet 5 µg/disc -- 4.6.4 Glucose 0.4 g/mL -- 4.6.5 G6P 1M: Glucose-6-Phosphate -- 4.6.6 HBT: 500 µM Histidine, 500µM Biotin, 500 µM Tryptophan Solution -- 4.6.7 Histidine HCl.H2O 5 mg/mL -- 4.6.8 KMg -- 4.6.9 MGA Plates -- 4.6.10 Minimal Glucose Master (MGM, MGMA and MGMAT) Plates -- 4.6.11 NADP 0.1 M -- 4.6.12 Nutrient Agar Plates -- 4.6.13 Nutrient Broth -- 4.6.14 Phosphate Buffer 0.2 M pH 7.4 -- 4.6.15 Positive Control and Diagnostic Mutagen Solutions -- 4.6.16 S9 Fraction -- 4.6.17 S9 Mix -- 4.6.18 Tetracycline 1 µg/disc -- 4.6.19 Top Agar Incomplete: TAI -- 4.6.20 Top Agar Complete: TAC -- 4.6.21 Tryptophan 5 mg/mL -- 4.6.22 VB Salts 50×: Vogel-Bonner Salts -- 4.7 Suggested Phases in Development of the Test -- 4.8 The Bacterial Strains -- 4.8.1 Genotypes of Routinely Used Strains -- 4.8.2 Obtaining the Tester Strains -- 4.8.3 Receipt of Bacterial Strains -- 4.8.4 Phenotyping of New Isolates. , 4.8.5 Freezing of Selected Isolates -- 4.8.6 Diagnostic Mutagen Test -- 4.9 Routine Testing -- 4.9.1 Designing a Study -- 4.9.1.1 Metabolic activation system -- 4.9.2 Test Article Considerations -- 4.9.2.1 Solvent selection -- 4.9.2.2 Dose volumes -- 4.9.2.3 Dose levels -- 4.9.3 Positive Controls -- 4.10 Standard Test Procedures -- 4.10.1 Plate Incorporation Method -- 4.10.2 Preincubation Method -- 4.10.3 Standard Study Design -- 4.10.4 Examination of the Plates -- 4.10.5 Interpretation of Results -- 4.10.5.1 Evaluation of toxicity -- 4.10.5.2 Validity of the study -- 4.10.5.3 Criteria for negative/positive/equivocal outcome -- 4.10.5.4 Unexpected and borderline results -- 4.10.6 Presentation of Results -- 4.10.7 Testing of Volatile and Gaseous Compounds -- 4.11 Screening Tests -- 4.11.1 Simplified Test Systems -- 4.11.2 Screening Tests Using Standard Tester Strains -- 4.11.3 Reduced Format Tests Using Standard Tester Strains -- 4.12 Appendix 1: Growing and Monitoring Suspension Cultures -- References -- 5 The Mouse Lymphoma TK Assay -- 5.1 Introduction -- 5.2 History -- 5.3 Provenance of the Cells -- 5.4 Spontaneous Mutation Frequency -- 5.5 Materials -- 5.5.1 Safety -- 5.5.2 Growth Medium -- 5.5.3 Cell Culture -- 5.5.4 Metabolic Activation -- 5.5.5 Test Item -- 5.5.6 Vehicle -- 5.5.7 Positive Controls -- 5.6 Study Design -- 5.6.1 General Test Conditions -- 5.6.2 Preliminary Toxicity Test -- 5.6.3 Main Mutation Test -- 5.6.3.1 Posttreatment procedures -- 5.6.3.2 Expression period -- 5.6.3.2.1 Expression period day 1 -- 5.6.3.2.2 Expression period day 2 -- 5.6.3.3 Viability assessment and mutant selection (microtiter version) -- 5.6.3.3.1 Colony counting (microtiter version) -- 5.6.3.4 Viability assessment and mutant selection (agar version) -- 5.6.3.4.1 Colony counting (agar version) -- 5.6.3.5 Analysis of results. , 5.6.3.5.1 Relative suspension growth -- 5.6.3.5.2 Toxicity assessment -- 5.6.3.5.3 MF assessment (microtiter version) -- 5.6.3.5.4 MF assessment (agar version) -- 5.6.3.6 Acceptance criteria -- 5.7 Evaluation Criteria -- 5.8 Predictivity of the MLA -- References -- 6 The In Vitro Micronucleus Assay -- 6.1 Introduction -- 6.2 Practical Considerations -- 6.2.1 Regulatory Guidelines -- 6.2.2 Good Laboratory Practice (GLP) -- 6.2.3 Cell Types -- 6.2.4 Laboratory Proficiency -- 6.2.5 Controls -- 6.2.6 Metabolic Activation -- 6.2.7 S9 Rat Liver Homogenate -- 6.2.8 Experimental Design -- 6.2.9 Cytotoxicity Measures -- 6.2.9.1 Methods used to determine cytotoxicity in the absence of cytochalasin B [16] (reproduced with permission of the author) -- 6.2.9.2 Method used to determine cytotoxicity in the presence of cytochalasin B -- 6.2.10 Historical Controls -- 6.3 Methods -- 6.3.1 Mononuclear Assay -- 6.3.2 Binuclear Assay -- 6.3.3 Centromeric Labeling -- 6.3.4 Nondisjunction Assay -- 6.4 Materials -- 6.4.1 Mononuclear Assay -- 6.4.2 Binuclear Assay -- 6.4.3 Centromeric Labeling -- 6.4.4 Nondisjunction Assay -- 6.5 Protocols -- 6.5.1 S9 Mix -- 6.5.2 Mononuclear Assay -- 6.5.2.1 Treatment schedules -- 6.5.2.2 Cell culture and treatment -- 6.5.2.3 Slide preparation -- 6.5.2.4 Coding of slides -- 6.5.2.5 Analysis of slides (microscope) -- 6.5.2.6 Analysis of slides (semiautomated scoring) -- Criteria for evaluation -- 6.5.3 Binuclear Assay -- 6.5.3.1 Treatment schedules -- 6.5.3.2 Human peripheral blood lymphocytes -- 6.5.3.3 Donors -- 6.5.3.4 Lymphocyte culture -- 6.5.3.5 Staining and analysis -- 6.5.3.6 Coding of slides -- 6.5.3.7 Analysis of slides -- 6.5.3.8 Evaluation of results (acceptance criteria and statistics) -- 6.5.3.9 Criteria for a valid assay -- 6.5.3.10 Evaluation of data -- 6.5.4 Centromeric Labeling. , 6.5.4.1 FISH using a programmable hotplate such as HYBrite™ or Thermobrite™ -- 6.5.4.2 Alterative protocol for FISH -- 6.5.4.3 Slide checking -- 6.5.4.4 Slide scoring -- 6.5.5 Nondisjunction Assay -- 6.5.5.1 FISH method -- 6.5.5.2 Slide checking -- 6.5.5.3 Slide scoring -- 6.6 Flow Cytometric Method -- 6.6.1 Equipment -- 6.6.2 Consumables -- 6.6.3 Reagents and recipes -- 6.6.4 Suspension Cell Protocol -- 6.6.5 Attachment Cell Protocol -- 6.6.6 Flow Cytometric Data Acquisition -- 6.6.7 Flow Cytometric Data Analysis -- 6.6.7.1 Independent cytotoxicity assessment -- 6.6.7.2 Criteria for a valid assay -- 6.6.7.3 Criteria for positive/negative outcomes -- 6.6.8 Creating an Analysis Template -- 6.6.9 Example Plate Layout -- 6.6.10 Example Results Table -- 6.6.11 Advice for Test Article Exposure -- 6.6.11.1 Suspension cells -- 6.6.11.2 Attachment cells -- 6.6.11.3 Metabolic activation -- 6.6.11.4 Positive controls -- 6.6.12 Use of Multichannel Aspirator with Bridge -- 6.6.13 Plate Placement During Nucleic Acid Dye B Photoactivation -- 6.6.14 Updates and Future Work -- References -- 7 The In Vitro Chromosome Aberration Test -- 7.1 Introduction -- 7.2 History -- 7.3 Fundamentals -- 7.4 Equipment -- 7.5 Consumables and Reagents -- 7.6 Reagents and Recipes -- 7.6.1 Colcemid 10 µg/mL in PBS -- 7.6.2 Fix -- 7.6.3 F-12 Complete -- 7.6.4 Freezing Medium 10% (CHO Cells) -- 7.6.5 Hypotonic Solution (0.075 M KCl) -- 7.6.6 Heparin Sodium 1000 U/mL -- 7.6.7 G6P 1 M: Glucose-6-Phosphate -- 7.6.8 KMg -- 7.6.9 NADP 0.1 M -- 7.6.10 PHA M Form (Phytohemagglutinin) -- 7.6.11 Phosphate Buffer 0.2 M, pH 7.4 -- 7.6.12 Positive Control Solutions -- 7.6.13 RPMI Complete -- 7.6.14 S9 Fraction -- 7.6.15 S9 Mix -- 7.7 Phases in Development of the Test -- 7.8 Cell Characterization -- 7.8.1 Modal Chromosome Number -- 7.8.2 Mycoplasma -- 7.8.3 Cell-Cycle Time -- 7.9 Routine Testing. , 7.9.1 General Considerations.

Additional Edition: ISBN 0-12-800764-8

Language: English