Biomarkers in Toxicology

Visit the link below to download the full version of this book:

https://medidownload.com/product/biomarkers-in-toxicology/

Click Download Now

xii FOREWORD

mitochondrial dysfunction and toxicity, and biomar- Greece, Hungary, India, Italy, Japan, Portugal, South
kers of oxidative and nitrosative stress. There are also Africa, and Spain. Professor Gupta deserves our
chapters on citrulline as a biomarker of diseases and gratitude for assembling such a distinguished group of
toxicities and pathological biomarkers in toxicology. experts to produce a comprehensive book on this
Part VIII is devoted to applications of biomarkers in rapidly growing and very important field.
toxicology with ten extremely comprehensive chapters.
Timothy C Marrs
The 64-chapter book has an outstanding array of
Edenbridge, UK
authors from the USA, Canada, Denmark, Finland,
 List of Contributors

Angela Aliberti Frank A. Barile

St. John’s University, College of Pharmacy and College of Pharmacy and Health Sciences, Department
Health Sciences, Department of Pharmaceutical of Pharmaceutical Sciences, Toxicology Division,
Sciences, Toxicology Division, Queens, NY, USA St. John’s University, Queens, NY, USA

Patrick Allard Sudheer R. Beedanagari

Department of Environmental Health Sciences and Bristol Myers Squibb, New Brunswick, NJ, USA
Institute for Society & Genetics, University of
California, Los Angeles, Los Angeles, CA, USA Enrico Bergamaschi
Department of Clinical and Experimental Medicine,
Arturo Anadón Occupational and Environmental Medicine Unit,
Department of Toxicology and Pharmacology, Faculty University of Parma, Parma, Italy
of Veterinary Medicine, Universidad Complutense
de Madrid, Madrid, Spain Sneha P. Bhatia
Research Institute for Fragrance Materials, NJ, USA
Vellareddy Anantharam
Parkinson’s Disorder Research Laboratory, Karyn Bischoff
Iowa Center for Advanced Neurotoxicology, Senior Extension Associate, Diagnostic Toxicologist,
Department of Biomedical Sciences, Iowa State Cornell University, New York State Animal Health
University, Ames, IA, USA Diagnostic Center, Ithaca, NY, USA

Anthony E. Archibong Vı́ctor Castellano

Department of Physiology, Meharry Medical College, Department of Toxicology and Pharmacology,
Nashville, TN, USA Faculty of Veterinary Medicine, Universidad
Complutense de Madrid, Madrid, Spain
Adam D. Aulbach
MPI Research, Mattawan, MI, USA Daniel T. Chang
US Environmental Protection Agency, National
Aryamitra Banerjee Exposure Research Laboratory, Research Triangle Park,
Toxicology Research Laboratory, University of Illinois NC, USA
at Chicago, Chicago, IL, USA
James J. Chen
Leah D. Banks Director of Biostatistics Branch, Division of
Department of Biochemistry & Cancer Biology, Meharry Bioinformatics and Biostatistics, National Center for
Medical College, Nashville, TN, USA Toxicological Research, US Food and Drug
Administration, Jefferson, AR, USA

xiii
xiv LIST OF CONTRIBUTORS

Deborah Citrin Jorge Estévez

Investigator, Radiation Oncology Branch, Instituto de Bioingenierı́a, Universidad Miguel
Center for Cancer Research, National Cancer Institute, Hernández de Elche, Spain
Bethesda, MD, USA
Daniel S. Fabricant
Rory Conolly Director, Division of Dietary Supplement Programs, Center
US Environmental Protection Agency, National for Food Safety and Applied Nutrition, US Food and Drug
Health and Environmental Effects Research Laboratory, Administration, College Park, MD, USA
Research Triangle Park, NC, USA
Anna M. Fan
R.W. Coppock Chief, Pesticide and Environmental Toxicology Branch,
Adjunct Professor, Department of Public Health Office of Environmental Health Hazard Assessment,
Sciences, School of Public Health University of Alberta, California Environmental Protection Agency,
Adjunct Professor, Environmental Health, Concordia Oakland, CA, USA
University College of Alberta, Edmonton, AB, Canada;
Ali Faqi
President and CEO, Robert W. Coppock, Toxicologist
Senior Director of Developmental and Reproductive
and Associates Ltd, Vegreville, AB, Canada
Toxicology, MPI Research, Mattawan, MI, USA
Lucio G. Costa Suzanne E. Fenton
Department of Environmental and Occupational Health National Toxicology Program (NTP) Laboratories
Sciences, University of Washington, Seattle, WA, USA Branch, Division of the NTP, National Institute of
Environmental Health Sciences, National Institutes
T.V. Damodaran of Health, Department of Health and Human Services,
Department of Biology, North Carolina Central Research Triangle Park, NC, USA
University, Durham, NC, USA
Vanessa A Fitsanakis
Kellye K. Daniels King University, Department of Biology, Bristol,
Director, Toxicology and Pathology, Southern Research TN, USA
Institute, Birmingham, AL, USA
John Flaskos
Curtis C. Dary Laboratory of Biochemistry and Toxicology,
US Environmental Protection Agency, National Faculty of Veterinary Medicine, Aristotle University
Exposure Research Laboratory, Las Vegas, NV, USA of Thessaloniki, Thessaloniki, Greece

Robin B. Doss Swaran J.S. Flora

Murray State University, Toxicology Department, Division of Regulatory Toxicology, Defence Research
Breathitt Veterinary Center, Hopkinsville, KY, USA and Development, Gwalior, MP, India

Filipe V. Duarte Sue M. Ford

Department of Life Sciences, Center for Neurosciences St. John’s University, College of Pharmacy & Health
and Cell Biology, University of Coimbra, Portugal Sciences, Pharmaceutical Sciences Department,
Toxicology Program, Jamaica, NY, USA
Margitta M. Dziwenka
Assistant Director of Operations, Health Sciences Domniki Fragou
Laboratory Animal Services, Faculty of Medicine and Laboratory of Forensic Medicine and Toxicology, School
Dentistry, University of Alberta, Edmonton, AB, of Medicine, Aristotle University of Thessaloniki, Greece
Canada; President and CEO, Toxalta Consulting Ltd.
Vegreville, AB, Canada Shayne C. Gad
Gad Consulting Services, Cary, NC, USA
Stephen Edwards
US Environmental Protection Agency, National Health Niels-Christian Ganderup
and Environmental Effects Research Laboratory, Chief Scientific Officer, Ellegaard Göttingen Minipigs
Research Triangle Park, NC, USA A/S, Dalmose, Denmark
 LIST OF CONTRIBUTORS xv

Dale R. Gardner Alan J Hargreaves

United States Department of Agriculture, Agricultural School of Science and Technology, Nottingham Trent
Research Service, Poisonous Plant Research Laboratory, University, Nottingham, UK
Logan, UT, USA
Kelly L. Harris
Ronette Gehring Department of Biochemistry & Cancer Biology, Meharry
Department of Anatomy and Physiology, College of Medical College, Nashville, TN, USA
Veterinary Medicine, Kansas State University,
Manhattan, KS, USA Holly E. Hatfield
Department of Biochemistry, University of Cincinatti
Fernando Gil Medical School, Cincinnati, OH, USA
Department of Legal Medicine and Toxicology,
Diane E. Heck
University of Granada School of Medicine,
Department of Environmental Health Science,
Granada, Spain
School of Health Sciences and Practice, New York
Medical College, Valhalla, NY, USA
Saryu Goel
Associate Director & Project Leader, Preclinical & Antonio F. Hernández
Competitive Intelligence, Supernus Pharmaceuticals, Department of Legal Medicine and Toxicology,
Inc. Rockville, MD, USA University of Granada School of Medicine,
Granada, Spain
Michael-Rock Goldsmith
US Environmental Protection Agency, National Corey J. Hilmas
Exposure Research Laboratory, Research Triangle Park, Chief, Dietary Supplements Regulation Implementation
NC, USA Branch, Division of Dietary Supplement Programs,
Center for Food Safety and Applied Nutrition, US Food
Christopher M. Grulke and Drug Administration, College Park, MD, USA
US Environmental Protection Agency, National
Exposure Research Laboratory, Research Triangle Park, Evangelia Hondroulis
NC, USA Nanobioengineering/Bioelectronics Laboratory,
Department of Biomedical Engineering, Florida
Mary Gulumian International University, Miami, FL, USA
Toxicology Department, National Institute
for Occupational Health, Hematology Darryl B. Hood
and Molecular Department of Toxicology, Division of Environmental Health Science, College
National Institute of Occupational Health, of Public Health, Department of Neuroscience, College
Johannesburg, South Africa of Medicine, The Ohio State University, Columbus,
OH, USA
P.K. Gupta
Patron and Founder, President, Society of Toxicology Kathryn Hudak
Founder, Editor-in-Chief, Toxicology International Research Fellow, Radiation Oncology Branch, Center for
President, Academy of Sciences for Animal Welfare, Cancer Research, National Cancer Institute, Bethesda,
Former Head of Div. Pharmacology & Toxicology, MD, USA
IVRI and Advisor to WHO, C- 44, Rajendra Nagar,
Bareilly, UP, India Pasi Huuskonen
School of Pharmacy/Toxicology, Faculty of Health
Ramesh C. Gupta Sciences, University of Eastern Finland, Kuopio, Finland
Professor and Head, Murray State University,
Toxicology Department, Breathitt Veterinary Center, Stewart B. Jacobson
Hopkinsville, KY, USA Shin Nippon Biomedical Laboratories USA, Ltd,
Everett, WA, USA
Sharon Gwaltney-Brant
Consultant, Veterinary Information Network, Huajun Jin
Adjunct Faculty, Department of Comparative Parkinson’s Disorder Research Laboratory, Iowa Center
Biosciences, College of Veterinary Medicine, University for Advanced Neurotoxicology, Department of Biomedical
of Illinois at Urbana-Champaign, IL, USA Sciences, Iowa State University, Ames, IA, USA
xvi LIST OF CONTRIBUTORS

Laurie B. Joseph Prasada Rao S. Kodavanti

Department of Pharmacology and Toxicology, Neurotoxicology Branch, Toxicity Assessment Division,
Rutgers University, Ernest Mario School of Pharmacy, NHEERL/ORD, US Environmental Protection Agency,
Piscataway, NJ, USA Research Triangle Park, NC, USA

Jun Kanno Urmila P. Kodavanti

Division of Cellular and Molecular Toxicology, Environmental Public Health Division, National
Biological Safety Research Center, National Institute Health and Environmental Effects Research Laboratory,
of Health Sciences, Tokyo, Japan Office of Research and Development, US Environmental
Protection Agency, Research Triangle Park, NC, USA
Anumantha G. Kanthasamy
Parkinson’s Disorder Research Laboratory, George A. Kontadakis
Iowa Center for Advanced Neurotoxicology, Department of Ophthalmology, Faculty of Medicine,
Department of Biomedical Sciences, Iowa State University of Crete, Heraklion, Greece
University, Ames, IA, USA
Péter Krajcsi
Arthi Kanthasamy SOLVO Biotechnology, Budaörs, Hungary
Parkinson’s Disorder Research Laboratory,
Iowa Center for Advanced Neurotoxicology, Gopala Krishna
Department of Biomedical Sciences, Iowa State Executive Director, Preclinical & Competitive
University, Ames, IA, USA Intelligence, Supernus Pharmaceuticals, Inc., Rockville,
MD, USA
Navinchandra M. Kaore
Department of Microbiology, People’s College Kavya A. Krishna
of Medical Sciences & Research Center, Bhopal, MP , Supernus Pharmaceuticals, Inc. Rockville, MD, USA
India
Maria Kummu
Shilpa N. Kaore Department of Pharmacology and Toxicology,
Department of Pharmacology, People’s College Institute of Biomedicine, and Center for Arctic
of Medical Sciences & Research Center, Bhopal, Medicine, Thule Institute of Oulu, University of Oulu,
MP, India Oulu, Finland

Bhupendra S. Kaphalia George D. Kymionis

Department of Pathology, University of Texas Medical Department of Ophthalmology, Faculty of Medicine,
Branch, Galveston, TX, USA University of Crete, Heraklion, Greece

Vesa Karttunen Michelle A. Lasher

School of Pharmacy/Toxicology, Faculty of Murray State University, Toxicology Department,
Health Sciences, University of Eastern Finland, Breathitt Veterinary Center, Hopkinsville, KY, USA
Kuopio, Finland
Chen-zhong Li
Greer E. Kaufman Nanobioengineering/Bioelectronics Laboratory,
Department of Bioanalysis, Southern Research Institute, Department of Biomedical Engineering, Florida
Birmingham, AL, USA International University, Miami, FL, USA

Ravneet Kaur Wei-Jiun Lin

Aveley, Western Australia Assistant Professor, Department of Applied
Mathematics, Feng Chia University, Taiwan
Hong Duck Kim
Department of Environmental Health Science, School of Bommanna G. Loganathan
Health Sciences and Practice, New York Medical Department of Chemistry and Watershed Studies
College, Valhalla NY, USA Institute, Murray State University, Murray, KY, USA
 LIST OF CONTRIBUTORS xvii

Jarkko Loikkanen Pushpinder K. Multani

School of Pharmacy/Toxicology, Faculty of Health Stress Neurobiology Division, Department of
Sciences, University of Eastern Finland, Kuopio, Finland Anesthesiology, Abramson Cancer Center, Children’s
Hospital of Philadelphia, Philadelphia, PA, USA
Anna E. Lokshin
University of Pittsburgh Cancer Institute, Hillman Päivi Myllynen
Cancer Center, Department of Medicine, Department Center for Arctic Medicine, Thule Institute, University
of Pathology, Department of Ob/Gyn, School of of Oulu, Oulu, Finland
Medicine, University of Pittsburgh, Pittsburgh,
PA, USA Kirsi Myöhänen
ECHA, European Chemicals Agency, Helsinki, Finland
Marcello Lotti
Professor of Medicine, Dipartimento di Scienze Rekek Negga
Cardiologiche, Toraciche e Vascolari, Universitá degli King University, Department of Biology, Bristol,
Studi Padova, Italy TN, USA

Tzu-Pin Lu Jairo Nelson

Postdoctoral Fellow, Division of Bioinformatics and Nanobioengineering/Bioelectronics Laboratory,
Biostatistics, National Center for Toxicological Research, Department of Biomedical Engineering, Florida
US Food and Drug Administration, Jefferson, AR, USA International University, Miami, FL, USA

Yi Lu Brian M. Nolen
Department of Pathology and Laboratory Medicine, University of Pittsburgh Cancer Institute, Hillman
University of Tennessee Health Science Center, Cancer Center, Pittsburgh, PA, USA
Memphis, TN, USA
Meliton N. Novilla
Chikezie Madu Shin Nippon Biomedical Laboratories USA, Ltd,
Department of Pathology and Laboratory Medicine, Everett, WA, USA; Adjunct Associate Professor of
University of Tennessee Health Science Center, Pathobiology, School of Veterinary Medicine, Purdue
Memphis, TN, USA University, West Lafayette, IN, USA

Rémi Magnan Stephanie Padilla

SOLVO Biotechnology, Budaörs, Hungary Integrated Systems Toxicology Division, National
Health and Environmental Effects Research Laboratory,
Brinda Mahadevan US Environmental Protection Agency, Research
Medical Safety & Surveillance, Abbott Laboratories, Triangle Park, NC, USA
Columbus, OH, USA
Carlos M. Palmeira
Jane A. Mantey Department of Life Sciences, Center for
Department of Neuroscience & Pharmacology, Meharry Neurosciences and Cell Biology, University of
Medical College, Nashville, TN, USA Coimbra, Portugal

Marı́a Rosa Martı́nez-Larrañaga Kip E. Panter

Department of Toxicology and Pharmacology, Faculty United States Department of Agriculture, Agricultural
of Veterinary Medicine, Universidad Complutense de Research Service, Poisonous Plant Research Laboratory,
Madrid, Madrid, Spain Logan, UT, USA

Vincent P. Meador Markku Pasanen

Covance Laboratories, Inc. Global Pathology, Madison, School of Pharmacy/Toxicology, Faculty of Health
WI, USA Sciences, University of Eastern Finland, Kuopio, Finland

Dejan Milatovic Daniel J. Patrick

Charlottesville, VA, USA Director of Pathology, MPI Research, Mattawan, MI, USA
xviii LIST OF CONTRIBUTORS

Sofia Pavanello Matthew R. Ricci

Research Scientist, Dipartimento di Scienze Research Diets, Inc., New Brunswick, NJ, USA
Cardiologiche, Toraciche e Vascolari, Universitá degli
Studi Padova, Italy Anabela P. Rolo
Center for Neurosciences and Cell Biology, University
Olavi Pelkonen of Coimbra, Department of Biology, University of
Department of Pharmacology and Toxicology, Aveiro, Portugal
Institute of Biomedicine, University of Oulu,
Oulu, Finland Magdalini Sachana
Laboratory of Biochemistry and Toxicology,
Claudia Pellacani Faculty of Veterinary Medicine, Aristotle University
Department of Neuroscience, University of Parma, of Thessaloniki, Thessaloniki, Greece
Parma, Italy
Heidi Sahlman
School of Pharmacy/Toxicology, Faculty of
Michael A. Pellizzon
Health Sciences, University of Eastern Finland,
Research Diets, Inc., New Brunswick, NJ, USA
Kuopio, Finland
Andrew D. Penman Nitin Saini
Vice President, Drug Development, Southern Research Janssen Research & Development, Malvern, PA, USA
Institute, Birmingham, AL, USA
William F. Salminen
Martin Phillips PAREXEL, Sarasota, FL, USA
US Environmental Protection Agency, National
Exposure Research Laboratory, Research Triangle Park, Kai Savolainen
NC, USA Nanosafety Research Centre, Finnish Institute of
Occupational Health, Helsinki, Finland
Jason Pitt
Northwestern University, Department of Physiology, Laura K. Schnackenberg
Chicago, IL, USA Division of Systems Biology, National Center
for Toxicological Research, US Food and Drug
Argyro Plaka Administration, Jefferson, AR, USA
Department of Ophthalmology, Faculty of Medicine,
University of Crete, Heraklion, Greece D.T. Selvam
Division of Microbiology, Division of Regulatory
Aramandla Ramesh Toxicology, Defence Research and Development,
Department of Biochemistry & Cancer Biology, Meharry Gwalior, MP, India
Medical College, Nashville, TN, USA
Praveen Sharma
Kausik Ray Professor and Head, Department of Biochemistry,
Scientific Review Officer, Scientific Review Branch, All India Institute of Medical Sciences, Jodhpur, India
Division of Extramural Activities, National
Institute on Deafness and Other Communication Qiang Shi
Disorders, National Institutes of Health, Bethesda, Division of Systems Biology, National Center for
MD, USA Toxicological Research, US Food and Drug
Administration, Jefferson, AR, USA
Casey E. Reed
National Toxicology Program (NTP) Laboratories Elina Sieppi
Branch, Division of the NTP, National Institute of Department of Pharmacology and Toxicology, Institute
Environmental Health Sciences, National Institutes of Biomedicine, and Center for Arctic Medicine, Thule
of Health, Department of Health and Human Services, Institute of Oulu, University of Oulu, Oulu, Finland
Research Triangle Park, NC, USA
Jon Sobus
Jenni Repo US Environmental Protection Agency, National
School of Pharmacy/Toxicology, Faculty of Health Exposure Research Laboratory, Research Triangle Park,
Sciences, University of Eastern Finland, Kuopio, Finland NC, USA
 LIST OF CONTRIBUTORS xix

Miguel A. Sogorb Kirsi Vähäkangas

Instituto de Bioingenierı́a, Universidad Miguel School of Pharmacy/Toxicology, Faculty of Health
Hernández de Elche, Spain Sciences, University of Eastern Finland, Kuopio, Finland

Melissa Stick Deon van der Merwe

Chief, Scientific Review Branch, Division of Extramural Kansas State University, Department of Diagnostic
Activities, National Institute on Deafness and Other Medicine/Pathobiology, Kansas State Veterinary
Communication Disorders, National Institutes of Diagnostic Laboratory, Manhattan, KS, USA
Health, Bethesda, MD, USA
Ana T. Varela
Markus Storvik Department of Life Sciences, Center for Neurosciences
School of Pharmacy/Toxicology, Faculty of Health and Cell Biology, University of Coimbra, Portugal
Sciences, University of Eastern Finland, Kuopio, Finland
Eugenio Vilanova
David T. Szabo Instituto de Bioingenierı́a, Universidad Miguel
National Center for Environmental Assessment, Office Hernández de Elche, Spain
of Research and Development, US Environmental
Protection Agency, Washington, DC, USA Suryanarayana V. Vulimiri
National Center for Environmental Assessment,
Yu-Mei Tan Environmental Protection Agency (EPA), Washington
US Environmental Protection Agency, National DC, USA
Exposure Research Laboratory, Research Triangle Park,
NC, USA Kevin D. Welch
United States Department of Agriculture, Agricultural
João S. Teodoro Research Service, Poisonous Plant Research Laboratory,
Department of Life Sciences, Center for Neurosciences Logan, UT, USA
and Cell Biology, University of Coimbra, Portugal
Xi Yang
Rogelio Tornero-Velez Division of Systems Biology, National Center for
US Environmental Protection Agency, National Toxicological Research, US Food and Drug
Exposure Research Laboratory, Research Triangle Park, Administration, Jefferson, AR, USA
NC, USA
Snjezana Zaja-Milatovic
Beáta Toth School of Medicine, University of Virginia,
SOLVO Biotechnology, Budaörs, Hungary Charlottesville, VA, USA

Aristides M. Tsatsakis Csaba K. Zoltani

Department of Forensic Sciences & Toxicology, Faculty Emeritus US Army Research Laboratory, Aberdeen
of Medicine, University of Crete, Heraklion, Greece Proving Ground, MD, USA
 C H A P T E R

1
Introduction
Ramesh C. Gupta

Biomarkers can be broadly defined as indicators or sig- some cases, as the metabolite of a drug can be used as a
naling events in biological systems or samples of mea- biomarker, and the drug and/or its metabolite has to be
surable changes at the molecular, biochemical, cellular, patented by the United States Patent Office and by a
physiological, pathological, or behavioral levels in similar governmental office/agency in other countries.
response to xenobiotics. The Biomarkers Definitions In the past, many drugs were developed with bio-
Working Group of the National Institutes of Health marker assays that guided their use, and this trend is
(NIH) has defined the biomarker as “a characteristic likely to continue in the future for drug discovery and
that is objectively measured and evaluated as an indica- development. With the judicious use of biomarkers, as
tor of normal biological processes or pharmacological in evidence-based medicine, patients are most likely to
responses to a therapeutic agent.” In the field of toxicol- benefit from select treatments and least likely to suffer
ogy, biomarkers have been classified as markers of from their adverse effects. On the contrary, utilization
exposure, effect, and susceptibility. Measurement of bio- of a bad biomarker can be as harmful to a patient as a
markers reflects the time-course of an injury and pro- bad drug. Therefore, biomarkers need to be validated
vides information on the molecular mechanisms of and evaluated by an accredited laboratory, which parti-
toxicity. These biomarkers provide us the confidence of cipates in a proficiency testing program, to provide a
accurate diagnosis, prognosis, and treatment. The bio- high level of confidence to both clinicians and patients.
markers of early chemical exposure can occur in concert In the toxicology field, biomarkers should be specific,
with biomarkers of early disease detection, and that accurate, sensitive, valid, biologically or clinically rele-
information aids in avoiding further chemical exposure vant, and easy and fast to perform in order to be useful
and in strategic development of a novel treatment, as predictive tools for toxicity testing and surveillance
including personalized medicine (i.e. treating the and for improving quantitative estimates of exposure
patient, and not the disease). In essence, with the utili- and dose. Therefore, biomarkers are utilized in biomo-
zation of specific biomarkers, an ounce of prevention nitoring data that are useful in a variety of applications,
can be worth a pound of treatment. from exposure assessment to risk assessment and
Biomarkers are used in drug development, during management.
preclinical and clinical trials, for efficacy and safety In the early 1990s, Dr. Maria Cristina Fossi, from the
assessment. Safety biomarkers can be used to predict, University of Siena, Italy, emphasized the approach for
detect, and monitor drug-induced toxicity during both the development and validation of nondestructive bio-
preclinical studies and human clinical trials. Developing markers over destructive biomarkers in the field of toxi-
highly sensitive methods and their validation for mea- cology. She described the ideal biomarker as being:
surement of biomarkers and understanding the resul- measurable in readily available tissues or biological pro-
tant data are complex processes that require a great ducts and obtainable in a noninvasive way; related to
deal of time, effort, and intellectual input. Furthermore, exposure and/or degree of harm to the organism; directly
understanding drug metabolism seems essential in related to the mechanism of action of the contaminants;

R. Gupta (Ed): Biomarkers in Toxicology. © 2014 Elsevier Inc. All rights reserved.
ISBN: 978-0-12-404630-6 DOI: http://dx.doi.org/10.1016/B978-0-12-404630-6.00001-4
3
4 1. INTRODUCTION

highly sensitive with techniques that require minimal invasive approach, and nondestructive biomarkers over
quantities of sample and are easy to perform and cost destructive biomarkers, but this may not be possible in
effective; and suitable for different species. The develop- all cases. Recently, the 2011 Joint SOT/EUROTOX-
ment and validation of new techniques in the laboratory Debate proposed that “biomarkers from blood and urine
may provide the basis for a valuable field method. But, will replace traditional histopathological evaluation to
before a new biomarker’s application, some basic infor- determine adverse responses,” identifying and compar-
mation is required, such as dose response relationships, ing the strengths and limitations of histopathology with
and biological and environmental factors, which can serum and urine biomarkers. Unlike histopathological
influence the baseline values of responses. It is important techniques, blood and urine biomarkers are noninvasive,
to mention that, when dealing with a biochemical or met- quantifiable, and of translational value. Of course, the
abolic biomarker, species differences can be the biggest complete replacement of histopathological biomarkers
challenge for any toxicologist. with blood and urine may not be possible in the near
Biomarkers have applications in all areas of toxicol- future, as in some instances histopathological biomarkers
ogy, especially in the field of pesticides, metals, myco- will still be used because of recent developments in
toxins, and drugs. In the case of veterinary toxicology, invaluable molecular pathology techniques.
biomarkers of plant toxins deserve equal attention. For the quest of developing the most sensitive and
Farmers, pesticide application workers, and greenhouse reliable biomarkers, integration of novel and existing
workers are exposed to pesticides by direct contact and biomarkers with a multidisciplinary approach appears
their family members can be exposed via second-hand fruitful. Furthermore, a multi-biomarkers approach
exposure. Measurement of residues of pesticides and seems more informative and accurate than a single bio-
their metabolites, and metals in urine, serves as the marker approach. The latest technologies, such as
most accurate and reliable biomarkers of exposure in microRNAs (miRNAs), have been well recognized as
agriculture, industrial, and occupational safety and reliable and robust biomarkers for early detection of
health settings. Recent evidence suggests that in utero or diseases, birth defects, pathological changes, cancer,
early life-exposure to certain pesticides, metals, and and toxicities. Because they are stable in biofluids, such
other environmental contaminants may cause neurode- as blood, there is rapidly growing interest in using
generative (Alzheimer’s, Parkinson’s, schizophrenia, miRNAs as diagnostic, prognostic, and predictive bio-
Huntington’s, ALS, and others), metabolic, and cardio- markers, and the outlook for the clinical application of
vascular diseases, and cancer later in life. In these dis- miRNA discoveries is promising, especially in molecu-
eases and many others, specific and sensitive lar medicine. The application of miRNAs is still very
biomarkers play important roles in early diagnosis, and new. Soon incorporating pharmacological and toxico-
this can serve as the cornerstone for timely therapeutic logical targeting of miRNAs into the development of
intervention. innovative therapeutic strategies will be routine. Still,
Mycotoxin related toxicity, carcinogenesis, and other more innovative biomarkers need to be developed that
health ailments are encountered in man and animals will be highly sensitive (biotechnology-based techni-
around the world. In developing countries, where regu- ques), require minimum quantities of sample, and will
latory guidelines are not strictly followed, adverse promise high throughput screening.
health effects (especially reproductive and developmen- At the recent annual meetings of the Society of
tal effects) are devastating. In these scenarios, early bio- Toxicology in San Francisco, CA, and San Antonio, TX,
markers of exposure play a pivotal role in avoiding USA, the EUROTOX in Stockholm, Sweden, and
further exposure to the contaminated food/feed and International Congress of Toxicology in Seoul, South
thus safeguard human and animal health. Korea, a large number of toxicologists emphasized the
With the current knowledge of system biology, meta- importance of biomarkers in health, disease, and toxic-
bonomics, and various mathematical and computa- ity. Accordingly, the Biomarkers in Toxicology book is
tional/chemometric modelings, undetectable biomarkers prepared to meet the challenges of today’s toxicologists,
can be discovered and these biomarkers can predict how pharmacologists, and physicians in academia, industry,
tissues respond to toxicants and drugs and/or their and government. This reference book may be of partic-
metabolites, and how the tissue damage and repair pro- ular interest to those in governmental agencies, such as
cesses compromise the tissue’s function. Imaging and USEPA, USFDA, NIOSH, OSHA, CDC, REACH, EFSA,
chemometric biomarkers are of greater sensitivity and etc. This is the most comprehensive biomarkers book to
carry more information than conventional biomarkers, as date as it covers every possible aspect of exposure,
they detect: (1) low levels of chemical exposure (expo- effects, and susceptibility to chemicals. There are many
sure biomarker), and (2) an early tissue response (endog- novel topics that are not covered in any previous book.
enous response biomarker). The priority will always be This book identifies and establishes the most sensitive,
for the development of a noninvasive approach over an accurate, unique, and validated biomarkers that can be
 1. INTRODUCTION 5

used as indicators of exposure and effect(s) of chemi- dedicated to the application of biomarkers in toxicol-
cals, and chemical-related long-term diseases, such as ogy, including the latest strategies and technologies in
cardiovascular, metabolic and neurodegenerative dis- the development of biomarkers, biomarkers in drug
eases, and cancer. Sixty-four chapters are organized development, safety evaluation, and toxicity testing
under eight sections with a user-friendly format, and and integration of biomarkers in biomonitoring of
each chapter is enriched with current literature and chemical exposure and risk assessment, especially in
references for further reading. The book begins with the context of environmental and occupational
general concepts of toxicity and safety testing and bio- medicine.
marker development using various animal and animal The editor remains indebted to the contributors of
alternative models, pharmacokinetic/PBPK modeling, this book for their hard work and dedication. These
followed by biomarkers of system/organ toxicity, che- contributors are highly qualified and considered author-
micals and biotoxins. There are several chapters on bio- ities in the fields of toxicology and pharmacology. He
markers of pharmaceuticals, nutraceuticals, petroleum expresses his gratitude to Ms. Denise Gupta for index-
products, chemical mixtures, radiation, nanoparticles, ing, and to Ms. Robin B. Doss and Ms. Michelle A.
epigenetics, genotoxicity, and carcinogenesis. Seven Lasher for their continuous support in technical assis-
chapters are devoted to special topics including bio- tance and text and reference checking. Finally, the edi-
markers of neurodegenerative diseases (Alzheimer’s tor would like to thank Ms. Mary Preap, Ms. Kristine
and Parkinson’s), mitochondrial dysfunction and toxic- Jones, Ms Marion Stockton, Ms Melissa Read and Mr
ity, oxidative/nitrosative stress, citrulline as a bio- Roderick Crews (Academic Press/Elsevier) for their
marker in diseases and toxicities, and pathological immense support at every stage of the production of
biomarkers. Lastly, a large number of chapters are this book.
 C H A P T E R

2
Rodents model for toxicity testing
and biomarkers
Shayne C. Gad

INTRODUCTION and in a manner that the relationship of findings in one

species as applying to another is known.
Accordingly, we will proceed to understand the
Three rodent species are widely used in toxicology: the
current uses of these three rodent species as predic-
rat, the mouse, and the hamster. Two of these, the rat
tive models for effects in humans, as well as how
and mouse, are the most widely used in experimental
they are measured and what their normal ranges
biology and medicine. These have formed the basis for
are.
the identification of toxicities associated with exposure
As these potential pieces of data are overviewed and
to drugs, industrial and agricultural chemicals, evalua-
considered, it is important to remember that each of
tion of their potential hazard, and understanding the
these biomarkers is a part of the overall picture as to
mechanisms of their toxicity since toxicology became an
what the model is predicting as per potential adverse
identified discipline.
effects in humans. Meaningful safety assessment
A large set of biomarkers are known for use in identi-
requires that all the data be incorporated in an inte-
fying and determining the relative (and relevant) risks
grated safety assessment.
to humans or other target species. These include:
The picture becomes both more complex but also
clearer as to relevance as new biomarkers are identified
G Body weights
and come to be understood. These include proteomics
G Hematology
(Amacher, 2010), new clinical chemistry parameters,
G Clinical chemistry
immune system responses, and real-time physiologic
G Organ weights
system measurements by telemetrized instrumentation
G Gross histologic changes at necropsy
(Gad, 2013).
G Microscopic
Table 2.1 (Gad, 2013) presents the classic associations
G Changes in physiologic functions and electro-
of biomarkers with renal and liver toxicity, while
physiology
Table 2.2 (adapted from Gad, 2013) presents an over-
view of the association between classical clinical chem-
Biomarkers are measurements of test model (animal)
istry parameters and specific target organ toxicities.
parameters that can provide important quantitative data
Table 2.3 summarizes causes associated with hemato-
about the biological state of the test model, predictive of
logical findings in the rat.
effects in humans. These biomarkers in toxicology are In the last few years, diligent efforts under the rubric
preferably shared by both test animals and humans, of the Critical Path Initiative have led to the

R. Gupta (Ed): Biomarkers in Toxicology. © 2014 Elsevier Inc. All rights reserved.
ISBN: 978-0-12-404630-6 DOI: http://dx.doi.org/10.1016/B978-0-12-404630-6.00002-6
7
8 2. RODENTS MODEL FOR TOXICITY TESTING AND BIOMARKERS

TABLE 2.1 Classic associations in toxicology

Liver Toxicity Renal Toxicity

Increased plasma activity of liver marker enzymes: e.g. alanine Increased water consumption and urine volume. Urine parameters may
and aspartate aminotransferases. change, e.g. enzymes and cellular debris.
Decreased plasma total protein concentration. Increased plasma concentrations of urea and creatinine. Proteinuria.
Increased coagulation times due to decreased synthesis of Severe renal toxicity may lead to decreased erythrocyte parameters due to
coagulation factors. effects on erythrocyte synthesis.
Increased liver weight due to enzyme induction or Increased kidney weight.
accumulation of lipid or glycogen.
Change in color or size at necropsy. Change in color or size at necropsy.
Histological findings such as necrosis or centrilobular Histological change, e.g. basophilic tubules or necrosis, papillary necrosis,
hypertrophy due to enzyme induction. or glomerular changes.

TABLE 2.2 Association of changes in biochemical parameters with actions at particular target organs

Parameter Blood Heart Lung Kidney Liver Bone Intestine Pancreas Notes
Albumin k k Produced by the liver; very significant reductions
indicate extensive liver damage
ALP m m m Elevations usually associated with cholestasis; bone
alkaline phosphatase tends to be higher in young
animals
ALT m Elevations usually associated with hepatic damage or
(formerly disease
SGPT)
AST (formerly m m m m Present in skeletal muscle and heart and most
SGOT) commonly associated with damage to them
Beta-2- m
microglobulin
Bilirubin m m Usually elevated due to cholestasis, due to either
(total) obstruction or hepatopathy
BUN m k Estimates blood filtering capacity of the kidneys; does
not become significantly elevated until the kidney
function is reduced to 60
75%
Calcium m Can be life threatening and result in acute death
Cholinesterase m k Found in plasma, brain, and RBC
CPK m Most often elevated due to skeletal muscle damage but
can also be produced by cardiac muscle damage; can be
more sensitive than histopathology
Creatinine m Also estimates blood filtering capacity of kidney as
BUN does
Glucose m Alterations other than those associated with stress
uncommon and reflect an effect on the pancreatic islets
or anorexia
GGT m Elevated in cholestasis; this is a microsomal enzyme,
and levels often increase in response to microsomal
enzyme induction
HBDH m m —
KIM-1 m
LDH m m m m Increase usually due to skeletal muscle, cardiac muscle,
or liver damage; not very specific
Protein (total) k k Absolute alterations usually associated with decreased
production (liver) or increased loss (kidney); can see
increase in case of muscle wasting (catabolism)
SDH mk Liver enzyme that can be quite sensitive but is fairly
unstable; samples should be processed as soon as
possible
Troponin m

Note: m, increase in chemistry values; k, decrease in chemistry values; ALP, alkaline phosphatase; BUN, blood urea nitrogen; CPK, creatinine phosphokinase;
GGT, gamma glutamyl transferase; HBDH, hydroxybutyric dehydrogenase; LDH, lactic dehydrogenase; RBC, red blood cells; SDH, sorbitol dehydrogenase;
SGOT, serum glutamic oxaloacetic transaminase (also called AST [aspartate amino transferase]); SGPT, serum glutamic pyruvic transaminase (also called ALT
[alanine amino transferase]).
 THE RAT 9

TABLE 2.3 Some probable conditions affecting hematological changes

Parameter Elevation Depression Parameter Elevation Depression

Red blood cells 1. Vascular shock 1. Anemias Platelets 1. Bone marrow
2. Excessive diuresis a) Blood Loss depression
3. Chronic hypoxia b) Hemolysis 2. Immune disorder
4. Hyperadreno- c) Low RBC
corticism production
Hematocrit 1. Increased RBC 1. Anemias Neutrophils 1. Acute bacterial
2. Stress 2. Pregnancy infections
3. Shock 3. Excessive hydration 2. Tissue necrosis
a) Trauma 3. Strenuous exercise
b) Surgery 4. Convulsions
4. Polycythemia 5. Tachycardia
6. Acute hemorrhage
Hemoglobin 1. Polycythemia 1. Anemias Lymphocytes 1. Leukemia
(increased 2. Lead Poisonings 2. Malnutrition
in production of 3. Viral infections
RBC)
Mean cell volume 1. Anemias 1. Iron deficiency Monocytes 1. Protozoal infections
2. B-12 deficiency
Mean corpuscular 1. Reticulocytosis 1. Iron deficiency Eosinophils 1. Allergy
hemoglobin 2. Irradiation
3. Pernicious anemia
4. Parasitism
White blood cells 1. Bacterial infections 1. Bone marrow Basophils 1. Lead poisoning
2. Bone marrow depression
stimulation 2. Cancer
chemotherapy
3. Chemical
intoxication
4. Splenic disorders

identification of a more specific set of clinical chemistry of cell membrane from cytoskeleton; stretching of the
biomarkers for key potential target organs, and these cell membrane results in increased membrane damage.
can be shown in this grouping: iv. Reactive Oxygen Species. Produced within the cell
and by infiltrating neutrophils and macrophages,
especially after restoration of blood flow to an area
Heart Troponins Zethelius et al. (2008)
(reperfusion injury). Cell injury triggers release of a
Kidney KIM-1, Albumin, Beta- Hoffmann et al. (2010);
2-Microglobulin Ozer et al. (2010); Vaidya
number of inflammatory cytokines and chemokines
et al. (2010) which amplify the host immune response and
Liver DILI (Drug Induced Shi et al. (2010) attract neutrophils to the site.
Liver Injury), ALT, BUN, v. Lipid Breakdown Products. Unesterified free fatty
coagulation factor acids, acyl carnitine, and lysophospholipids. These
have a detergent effect on membranes and may
exchange with membrane phospholipids, causing
Other recently identified biomarkers for specific tar-
permeability changes.
gets include:

i. Mitochondrial Dysfunction. Increased uptake of cal-

cium (because of ATP depletion) by mitochondria
activates phospholipases, resulting in accumulation THE RAT
of free fatty acids. These cause changes in the per-
meability of mitochondrial membranes, such as the
mitochondrial permeability transition.
Use in toxicological research
ii. Progressive Loss of Phospholipids. Increased degrada- Ideally, safety testing of products intended for use in
tion by endogenous phospholipases and inability of humans, or to which humans could be exposed, should
the cell to keep up with synthesis of new phospholi- be done in humans. The data from humans would
pids (reacylation, an ATP-dependent process). apply without reservation to complex human physiol-
iii. Cytoskeletal Abnormalities. Activated proteases lyse cyto- ogy and cellular/biochemical mechanisms and human
skeletal elements and cell swelling causes detachment risk assessment. Unfortunately, humans cannot be used
10 2. RODENTS MODEL FOR TOXICITY TESTING AND BIOMARKERS

for this purpose. Therefore, the choice of an appropriate incidence of spontaneous glomerular sclerosis (Bolton
species for toxicology studies should be based upon a et al., 1976), sensitivity to the carcinogenic actions of
comparison of the pharmacokinetics and metabolism of 7,12-dimethylbenz(a)anthracene (Boyland and Sydnor,
the test compound in different laboratory species and 1962), the effects of trimethyltin on operant behavior
man. In the absence of this data this choice is often and hippocampal GFAP (MacPhail et al., 2003), differ-
based upon practicality and economics. The rat has ences in renal carcinogenesis (Hino et al., 2003), differ-
become a species of choice because of metabolic similar- ences in cytochrome P4501A1 gene expression caused
ities, as well as their small size, relatively docile nature, by 2,3,7,8-tetrachlorodibenzo-p-dioxin in the liver (Jana
short life span, and short gestation period. The exten- et al., 1998), susceptibility to 4-nitroquinoline 1-oxide
sive use of the rat in research has led to the develop- inducing carcinoma (Kitano et al., 1992), and differences
ment of a large historical database of their nutrition, in the levels of drug-metabolizing enzymes (Page and
diseases, and general biology. Vesell, 1969). In recent years, research and breeding
programs have been focused on producing inbred and
Characteristics outbred strains focused on specific disease models and
susceptibility to the development of certain carcinoma.
While the rat is a species of choice in toxicology research When choosing a strain for use, it is important to con-
because of the many physiological similarities and ana- sider these differences.
tomical characteristics, differences exist that must be Of importance for carcinogenicity studies, strain dif-
considered when designing and conducting studies with ferences have been found in the incidence of spontane-
this animal. Rats are obligate nose breathers, and, as ous tumors. Table 2.4 gives the incidence of
such, an inhaled test material is subject to nasal filtration spontaneous tumors found in commonly used strains in
and absorption. The placenta is considerably more carcinogenicity studies. The historical incidence is
porous in the rat. This difference may increase the important to the analysis of a study in that a high spon-
chance of fetal exposure to an administered test material taneous rate may mask a small test material related
or increase the overall level of fetal exposure to an increase in tumor incidence.
administered test material. The overall distribution of Due to lower spontaneous tumor rates, the Wistar has
intestinal microflora is different in the rat, which may become the most popular strain in toxicological research.
lead to differences in the metabolism of an orally admin-
istered test material. These and other differences in the Normal physiological values
rat may lead to positive signs of toxicity to a test material
that may not be present in a different species. General values for selected physiological parameters are
given in Tables 2.5 and 2.6. Normal values will vary
based upon the strain of animal, supplier, feed, and
Strain differences
housing conditions. These tables should be used as a
Breeding rats for specific characteristics has produced point of reference only.
some physiological differences between strains of rats.
Some of these differences are known to affect how the
various strains react to toxicants. Among others, strain-
Study designs
specific differences have been found in sensitivity to
thiourea (Dieke and Richter, 1945), sensitivity to acet- The length and design of toxicology studies used to pre-
aminophen nephrotoxicity (Newton et al., 1985a,b), the dict human risk are governed by guidelines issued by

TABLE 2.4 Incidence of common spontaneous tumors in Fischer 344 and CD (SD)IGS rats

% Tumors in Untreated Rats

CD(SD)IGS CD (SD) Fisher Wistar

Organ Tumor Type Male Female Male Female Male Female Male Female
Adrenal gland Pheochromocytoma 10.0 2.3 11.3 2.3 11.9 3.2 3.2 1.3
Mammary gland Fibroadenoma 1.4 44.5 1.3 16.7 0.8 7.1 1.2 30.2
Pancreas Islet cell adenoma 3.6 1.4 4.0 0.3 1.5 0.2 5.3 1.9
Pituitary Gland Adenoma pars distalis 33.6 56.8 35.7 50.3 12.4 28.2 41.1 65.8
Testis Interstitial cell tumor 1.8 7.0 74.6 4.3
Thyroid gland C-Cell Adenoma 10.5 5.0 5.0 5.7 12.5 8.2 10.1 10.7

Source: Adapted from Charles River (2000), Mitsumori et al. (2001)