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Point Mutation Detection in the Cotton Rat. Christine H Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 2000 Indices of Exposure to Polycyclic Aromatic Hydrocarbons in a Bioindicator Species: Point Mutation Detection in the Cotton Rat. Christine H. Lemarchand-carpentier Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Lemarchand-carpentier, Christine H., "Indices of Exposure to Polycyclic Aromatic Hydrocarbons in a Bioindicator Species: Point Mutation Detection in the Cotton Rat." (2000). LSU Historical Dissertations and Theses. 7277. https://digitalcommons.lsu.edu/gradschool_disstheses/7277 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6* x 9” black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. Bell & Howell Information and Learning 300 North Zeeb Road, Ann Arbor, Ml 48106-1346 USA 800-521-0600 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. INDICES OF EXPOSURE TO POLYCYCLIC AROMATIC HYDROCARBONS IN A BIOINDICATOR SPECIES: POINT MUTATION DETECTION IN THE COTTON RAT A Dissertation Submitted to the Graduate Faculty o f the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Interdepartmental Program in Veterinary Medical Sciences through The Department of Veterinary Physiology, Pharmacology and Toxicology by Christine Lemarchand-Carpentier DEDV, University of Nantes, France, 1993 August, 2000 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number 9984345 UMI* UMI Microform9984345 Copyright 2000 by Bell & Howell Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. Bell & Howell Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DEDICATION These years o f efforts and sacrifices are dedicated to my family, in particular, those for whom there will be no opportunity to make up our absence. The Atlantic Ocean represented a gap in a lot of hearts. Nonetheless, my status of expatriated student was accepted with good will. ii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ACKNOWLEDGMENTS This research was supported by grants from The Department of Energy and The State of Louisiana Board of Regents. I would like to thank my committee members: Dr Jay Means, who gave me the opportunity to enter this program; Dr Vince Wilson, for his help and optimism in the development of PCR/RE/LCR and the use of his laboratory and equipment; Dr Steven Barker, for his advice and the use of its powerful computer!; and Dr Steven Nicholson. This is also the opportunity to acknowledge Dr Beth Goad, for her advice in pathology and her logistic support, Dr David Baker for communicating his experimental results on F344 rats, and Dr Leberg and his assistant for trapping wild Cotton rats. Thank you to Laynette Spring, Vladimir and Suzan Newman for their technical advice in molecular biology, July Millard and all thermocycler users for your comprehension, while I was monopolizing this piece of equipment, Xuiquin Yin and Kristin Clark for avoiding me excessive commuting on the campus, and Drs Melanie Rembert and Rett Stout for their help in handling and procedures on laboratory rodents. The expertise of Michael Hooks from the Office of Environmental and Occupational Safety was of great help in the handling of hazardous substances. Staff from the Veterinary Computer Services kindly provided their time and advice to resolve my computer problems. The assistance in administrative matters and the humor o f the secretaries from the Departments of Veterinary Pathology and Veterinary Physiology, Pharmacology and Toxicology were also appreciated during this project. Last but not least, merci to Thomas, my husband, for his support and the symbiosis we were able to achieve during these years, particularly when our little one, Audrey, was sick. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE OF CONTENTS Dedication.............................................................................................................................................ii Acknowledgments ................................................................................................................................iii Abbreviations....................................................................................................................................... vi Abstract........................... viii Chapter I: Introduction....................................................................................................................... 1 References ..................................... 5 Chapter 2: Literature Review........................................................................................................... 10 Human Exposure to Environmental Pollutants .......................... 10 Polycyclic Aromatic Hydrocarbons ......................................... 10 Exposure in Occupational Settings ........................................................ 13 Exposure in the General Population ..................................................................................... 14 2-Aminoanthracene and Benzo[a]anthracene, Models of PAHs and Substituted PAHs 16 2 Aminoanthracene .................................................................................................................16 Benz[a]anthracene................................................................................................................. 23 Monitoring o f Environmental Pollutants Using Animals ............... 29 Historical Examples .............................................................................................. 29 Definitions .............................................................................................................................. 30 Limitations and Advantages of Biomonitoring ................................................................. 32 The Case o f PAHs................................................................................................................. 33 Cotton Rats ( Sigmodon hispidus) as Candidates ................................................................ 34 The Process o f Carcinogenesis ....................................................................... 38 Factors Influencing Fixation Mutation at One S ite ............................................................39 Factors Influencing the Potential Biological Effect of a Mutation ....................................45 Mutation Spectrum ....................................................................................... 54 Below the Threshold Mutation Detection Assays ......................................................................56 References ......................................................................................................................................58 Chapter 3: Evaluation of the Effects of 2-Aminoanthracene and Benz(a|anthracene in Cotton Rats (Sigmodon hispidus) 78 Introduction ....................................................................................................................................78 Experimental Design .....................................................................................................................80 Materials ............ 80 Animals _____ ...................... 80 Safety Measures.................................................................................................................... 8 1 Preliminary Study: Dietary Exposure o f Cotton Rats .........................................................81
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