Drug Safety Evaluation Methods and Protocols by Gautier, Jean-Charles (Ed.)

Drug Safety Evaluation

Methods and Protocols

Series: Methods in Molecular Biology, Vol. 691

Gautier, Jean-Charles (Ed.)

2011, XIII, 431p. 78 illus., 4 illus. in color.

-Features a compendium of analytical technologies, including some review chapters, with a focus on clarity and applicability in real life laboratory practice
-Contains methods and techiques easily translated to a myriad of scientific disciplines
-Presents key tips from the experts on the best way to conduct these techniques succesfully

Non-clinical drug safety evaluation, the assessment of the safety profile of therapeutic agents through the conduct of laboratory studies in in vitro systems and in animals, is an essential step in the progress of new pharmaceuticals heading toward the ultimate goal of clinical trials and, eventually, approval. In Drug Safety Evaluation: Methods and Protocols, expert researchers detail a compendium of analytical technologies with a focus on clarity and applicability in real life laboratory practice. These meticulous contributions feature key topics such as acute to chronic general toxicity studies, histopathology studies, reproductive toxicity studies, genotoxicity studies, safety pharmacology studies, investigative toxicity studies, and safety biomarker studies. As a volume in the highly successful Methods in Molecular Biology™ series, chapters include brief introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Drug Safety Evaluation: Methods and Protocols serves as an ideal guide to this field, helpful to pharmaceutical scientists, toxicologists, biochemists, and molecular biologists as well as scientists from all other disciplines who wish to translate these thorough methods into their own work.

Content Level » Professional/practitioner

Keywords » Carcinogenicity - Chemical protein adducts - Developmental toxicity - Experimental design - General toxicity studies - Genotoxicity - Novel safety biomarkers - Reproductive toxicology - Safety pharmacology - Screening assays - Therapeutic agents

Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

PART I GENERAL TOXICOLOGY

1 Developing Combination Drugs in Preclinical Studies. . . . . . . . . . . . . . . . . . . . . . 3
Alberto Lodola

2 Preclinical Evaluation of Juvenile Toxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Paul C. Barrow, Stéphane Barbellion, and Jeanne Stadler

PART II PATHOLOGY

3 Necropsy and Sampling Procedures in Rodents. . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Laurence Fiette and Mohamed Slaoui

4 Histopathology Procedures: From Tissue Sampling to Histopathological
Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Mohamed Slaoui and Laurence Fiette

5 Principles and Methods of Immunohistochemistry . . . . . . . . . . . . . . . . . . . . . . . . 83
José A. Ramos-Vara

6 Tissue Microarrays and Digital Image Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
Denise Ryan, Laoighse Mulrane, Elton Rexhepaj,
and William M. Gallagher

PART III GENETIC TOXICOLOGY

7 Micronucleus Assay and Labeling of Centromeres with FISH Technique. . . . . . . . 115
Ilse Decordier, Raluca Mateuca, and Micheline Kirsch-Volders

8 The Use of Bacterial Repair Endonucleases in the Comet Assay. . . . . . . . . . . . . . . 137
Andrew R. Collins
PART IV SAFETY PHARMACOLOGY

9 Manual Whole-Cell Patch-Clamping of the HERG Cardiac K+ Channel . . . . . . . . 151
Xiao-Liang Chen, Jiesheng Kang, and David Rampe

PART V INVESTIGATIVE TOXICOLOGY

10 Generation and Analysis of Transcriptomics Data . . . . . . . . . . . . . . . . . . . . . . . . . 167
Philip D. Glaves and Jonathan D. Tugwood

11 Protocols of Two-Dimensional Difference Gel Electrophoresis
to Investigate Mechanisms of Toxicity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Emmanuelle Com, Albrecht Gruhler, Martine Courcol,
and Jean-Charles Gautier

12 Protocols and Applications of Cellular Metabolomics in Safety Studies
Using Precision-Cut Tissue Slices and Carbon 13 NMR . . . . . . . . . . . . . . . . . . . . 205
Gabriel Baverel, Sophie Renault, Hassan Faiz, Maha El Hage,
Catherine Gauthier, Agnès Duplany, Bernard Ferrier, and Guy Martin

13 Statistical Analysis of Quantitative RT-PCR Results. . . . . . . . . . . . . . . . . . . . . . . . 227
Richard Khan-Malek and Ying Wang

14 Evaluation of Mitochondrial Respiration in Cultured Rat Hepatocytes . . . . . . . . . 243
Jean-Pierre Marchandeau and Gilles Labbe

PART VI SCREENING ASSAYS FOR DEVELOPMENTAL TOXICITY

15 FETAX Assay for Evaluation of Developmental Toxicity . . . . . . . . . . . . . . . . . . . . 257
Isabelle Mouche, Laure Malesic, and Olivier Gillardeaux

16 Evaluation of Embryotoxicity Using the Zebrafish Model . . . . . . . . . . . . . . . . . . . 271
Lisa Truong, Stacey L. Harper, and Robert L. Tanguay

PART VII CHEMICAL PROTEIN ADDUCTS

17 Protocols of In Vitro Protein Covalent Binding Studies in Liver . . . . . . . . . . . . . . 283
Jean-François Lévesque, Stephen H. Day, and Allen N. Jones

18 Utilization of MALDI-TOF to Determine Chemical-Protein Adduct
Formation In Vitro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
Ashley A. Fisher, Matthew T. Labenski, Terrence J. Monks,
and Serrine S. Lau

19 Utilization of LC-MS/MS Analyses to Identify Site-Specific
Chemical Protein Adducts In Vitro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 317
Ashley A. Fisher, Matthew T. Labenski, Terrence J. Monks,
and Serrine S. Lau

20 One-Dimensional Western Blotting Coupled to LC-MS/MS
Analysis to Identify Chemical-Adducted Proteins in Rat Urine . . . . . . . . . . . . . . . 327
Matthew T. Labenski, Ashley A. Fisher, Terrence J. Monks,
and Serrine S. Lau

21 Identification of Chemical-Adducted Proteins in Urine
by Multi-dimensional Protein Identification Technology
(LC/LC–MS/MS). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
Matthew T. Labenski, Ashley A. Fisher, Terrence J. Monks,
and Serrine S. Lau

PART VIII SAFETY BIOMARKERS

22 Optimization of SELDI for Biomarker Detection in Plasma . . . . . . . . . . . . . . . . . 351
Jean-Francois Léonard, Martine Courcol, and Jean-Charles Gautier

23 Differential Proteomics Incorporating iTRAQ Labeling
and Multi-dimensional Separations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
Ben C. Collins, Thomas Y.K. Lau, Stephen R. Pennington,
and William M. Gallagher

24 NMR and MS Methods for Metabonomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 385
Frank Dieterle, Björn Riefke, Götz Schlotterbeck, Alfred Ross,
Hans Senn, and Alexander Amberg

25 Absolute Quantification of Toxicological Biomarkers by Multiple
Reaction Monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 417
Thomas Y. K. Lau, Ben C. Collins, Peter Stone, Ning Tang,
William M. Gallagher, and Stephen R. Pennington

Index