Molecular Toxicology Protocols

ISBN-13: 9781493956548 / Angielski / Miękka / 2016 / 646 str.

Phouthone Keohavong; Stephen G. Grant

Opis

This book details techniques for analyzing genotoxic exposure and the resulting biological effects, including intermediate biomarkers such as DNA and chromosomal damage, mutations in reporter and oncogenes, and the earliest possible detection of cancer.

Spis treści

Part I. Toxicoproteomics

 

1. Array-Based Immunoassays with Rolling-Circle Amplification Detection

Katie Partyka, ShuangshuangWang, Ping Zhao, Brian Cao, and Brian Haab

 

2. Analysis of Protein Changes Using Two Dimensional Difference Gel Electrophoresis

Weimin Gao

 

 

Part II. Toxicant-Induced Inflammation

 

3. Assessment of Pathological and Physiological Changes in Mouse Lung Through Bronchoalveolar Lavage

Yuanpu Peter Di

 

4. Analysis of Clinical and Biological Samples Using Microsphere-Based Multiplexing Luminex System

Yingze Zhang, Rahel Birru, and Y. Peter Di

 

           

Part III. Gene Promoter Methylation

 

5. Detection of DNA Methylation by MeDIP and MBDCap Assays: An Overview of Techniques

Hang-Kai Hsu, Yu-I Weng, Pei-Yin Hsu, Tim H.-M. Huang, and Yi-Wen Huang

 

6. Screening of DNA Methylation Changes by Methylation-Sensitive Random Amplified Polymorphic DNA-Polymerase Chain Reaction (MS-RAPD-PCR)

Kamaleshwar P. Singh

 

 

Part IV. Array Technologies

 

7. Strategies for Measurement of Biotransformation Enzyme Gene Expression

Marjorie Romkes and Shama C. Buch

 

8. Genotyping Technologies: Application to Biotransformation Enzyme Genetic Polymorphism Screening

Marjorie Romkes and Shama C. Buch

 

9. TaqMan Fluorogenic Detection System to Analyze Gene Transcription in Autopsy Material

            Kaori Shintani-Ishida, Bao-Li Zhu, and Hitoshi Maeda

 

 

Part V. Analysis of DNA Adducts

 

10. 32P-Postlabelling Analysis of DNA Adducts

            David H. Phillips and Volker M. Arlt

 

11. Modification of the 32P-Postlabeling Method to Detect a Single Adduct Species as a Single Spot

            Masako Ochiai, Takashi Sugimura, and Minako Nagao

 

12. DNA Isolation and Sample Preparation for Quantitation of Adduct Levels by Accelerator Mass Spectrometry

Karen H. Dingley, Esther A. Ubick, John S. Vogel, Ted J. Ognibene, Michael A. Malfatti, Kristen Kulp, and Kurt W. Haack

 

13. Analysis of DNA Strand Cleavage at Abasic Sites

            Walter A. Deutsch and Vijay Hegde

 

 

Part VI. Detection of Chromosomal and Genome-Wide Damage

 

14. Premature Chromosome Condensation in Human Resting Peripheral Blood Lymphocytes without Mitogen Stimulation for Chromosome Aberration Analysis Using Specific Whole Chromosome DNA Hybridization Probes

            Rupak Pathak and Pataje G. S. Prasanna

 

15. Mutagen Sensitivity as Measured by Induced Chromatid Breakage as a Marker of Cancer Risk

            Xifeng Wu, Yun-Ling Zheng, and T. C. Hsu

 

16. Pulsed-Field Gel Electrophoresis Analysis of Multicellular DNA Double-Strand Break Damage and Repair

            Nina Joshi and Stephen G. Grant

 

 

Part VII. Detection and Characterization of Surrogate Gene Mutation

 

17. Detection of Pig-a Mutant Erythrocytes in the Peripheral Blood of Rats and Mice

            Vasily N. Dobrovolsky, Xuefei Cao, Javed A. Bhalli, and Robert H. Heflich

 

18. The Blood-Vased Glycophorin A Human In vivo Somatic Mutation Assay

            Nicole T. Myers and Stephen G. Grant

 

19. Flow Cytometric Quantification of Mutant T Cells with Altered Expression of the T-Cell Receptor: Detecting Somatic Mutants in Humans and Mice

            Seishi Kyoizumi, Yoichiro Kasunoki, and Tomonori Hayashi

 

20. Analysis of in Vivo Mutation at the HPRT and TK Genes of Mouse Lymphocytes

            Vasily N. Dobrovolsky, Joseph G. Shaddock, and Robert H. Heflich

 

21. Quantifying In Vivo Somatic Mutations Using Transgenic Mouse Model Systems

Roy R. Swiger

 

22. The Human T-Cell Cloning Assay: Identifying Genotypes Susceptible to Drug Toxicity and Somatic Mutation

Sai-Mei Hou

 

23. Molecular Analysis of HPRT Mutations in Human Lymphocytes

            Phouthone Keohavong, Liqiang Xi, Stephen G. and Grant

 

24. Simultaneous Quantification of t(14;18) and HPRT Exon 2/3 Deletions in Human Lymphocytes

            James C. Fuscoe

 

 

Part VIII. Detection and Characterization of Cancer Gene Mutation

 

25. Mutation Screening of the TP53 Gene by Temporal Temperature Gel Electrophoresis

            Therese Sørlie, Hilde Johnson, Phuong Vu, Guro Elisabeth Lind, Ragnhild Lothe, and Anne-Lise Børreson-Dale

 

26. Detection of Point Mutations of K-ras Oncogene and p53 Tumor Suppressor Gene in Sputum Samples

Wei-Min Gao and Phouthone Keohavong

 

27. ACB-PCR Quantification of Somatic Oncomutations

Meagan B. Myers, Page B. McKinzie, Yiying Wang, Fanxue Meng, and Barbara L. Parsons

 

28. Gel-Based Nonradioactive Single Strand Conformational Polymorphism and Mutation Detection

Vibhuti Gupta, Reetakshi Arora, Sailesh Gochhait, Narendra K. Bairwa, and Rameshwar N. K. Bamezai

 

29. Detection and Characterization of Oncogene Mutations in Preneoplastic and Early Neoplastic Lesion

            Toshinari Minamoto

 

30. Detection of DNA Double-Strand Breaks and Chromosome Translocations Using Ligation-Mediated PCR and Inverse PCR

            Sheetal Sing, Shyh-Jen Shih, and Andrew T. M. Vaughan

 

 

Part IX. Analysis of DNA Damage and Repair Mechanisms

 

31. Quantitative PCR-Based Measurement of Nuclear and Mitochondrial DNA Damage and Repair in Mammalian Cells

            Amy Furda, Janine H. Santos, Joel N. Meyer, and Bennett Van Houten

 

32. The Sister Chromatid Exchange (SCE) Assay

Dawn M. Stults, Michael W. Killen, and Andrew J. Pierce

 

33. The Gene Cluster Instability (GCI) Assay for Recombination

Michael W. Killen, Dawn M. Stults, and Andrew J. Pierce

 

34. Measuring Recombination Proficiency in Mouse Embryonic Stem Cells

            Andrew J. Pierce and Maria Jasin

 

35. Microsatellite Instability: An Indirect Assay to Detect Defects in the Cellular Mismatch Repair Machinery

            Narendra K. Bairwa, Anjana Saha, Sailesh Gochhait, Ranjana Pal, Vibhuti Gupta, and Rameshwar N. K. Bamezai

 

36. Unscheduled DNA Synthesis: The Clinical and Functional Assay for Global Genomic Nucleotide Excision Repair

            Jean J. Latimer, and Carytal M. Kelly

 

37. An Immunoassay for Measuring Repair of UV Photoproducts

            Shirley McCready

 

38. Analysis of Double-Strand Break Repair by Non-Homologous DNA End Joining in Cell-Free Extracts from Mammalian Cells

            Petra Pfeiffer, Andrea Odersky, Wolfgang Goedecke, and Steffi Kuhfittig-Kulle

 

 

Part X. Analysis of Cellular Bioenergetics and Apoptosis

 

39. Bioenergetic Analysis of Intact Mammalian Cells Using the Seahorse XF42 Extracellular Flux Analyzer and a Luciferase ATP Assay

            Michelle Barbi de Moura, and Bennett Van Houten

 

40. Quantification of Selective Phosphatidylserine Oxidation During Apoptosis

            James P. Fabisiak, Yulia Y. Tyurina, and Valerian E. Kagan

 

41. Quantitative Method of Measuring Phophatidylserine Externalization During Apoptosis Using Electron Paramagnetic Resonance (EPR) Spectroscopy and Annexin-Conjugated Iron

            James P. Fabisiak, Grigory G. Borisenko, and Valerian E. Kagan

 

42. Detection of Programmed Cell Death in Cells Exposed to Genotoxic Agents Using a Caspase Activation Assay

Madhu Gupta, Madhumita Santra, and Patrick P. Koty

Blurb

Molecular Toxicology Protocols, Second Edition aims to bring together a series of articles describing validated methods to elucidate specific molecular aspects of toxicology, the emphasis being on the application of molecular methods to genetic toxicology. The volume is divided into ten parts, roughly corresponding to the spectrum of biomarkers intermediate between exposure and disease outcomes as proposed in molecular epidemiology models. Subjects of these new chapters range from preparation of fluid specimens for analysis of cellular inflammatory responses to genotoxic insults to sensitive methods for proteomic analysis and aberrant DNA methylation patterns. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Molecular Toxicology Protocols, Second Edition addresses not only the needs of molecular biologists and toxicologists, but also those of individuals interested in applying molecular methods to clinical applications, such as geneticists, pathologists, biochemists, and epidemiologists.