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Department of Pathology UMI Number: 9526054 UMI Microform 9526054 Copyright 1995, by OMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, MI 48103 ACKNOWLEDGMENTS I wish to thank Dr. Hari M. Sharma, research adviser, for his ideas, insights, and inspiration, and Dr. John A. Lott, academic adviser, for his guidance, patience, and encouragement. I am very grateful to the members of my graduate research committee, Drs. Peter B. Baker and Rao V. Panganamala for their suggestions and assistance. I would like to acknowledge the help of Ellen Kauffman, Jim Favret, Michael Burton and Drs. Howard Newman, Atef Hanna and John W. Heinz for their support and technical assistance in this study. I also would like to thank Dr. John B. Brandt, and Ms. Kathy Reger-Heinz for their generosity in permitting me to use the instruments and reagents in the clinical laboratories. I also wish to thank my wife, Kuem Ja, who has not only offered me technical help, but also love, patience, prayer, and encouragement, and my daughter, Susie for her sacrifice. VITA October 23,1938 .............................. Born - Hongsong, Choong Nam, Korea 1959 - 1960 ........................................ Served in Korean Army February, 1963...................................B. Sc. in Biology, The Yonsei University, Seoul, Korea 1963 - 1967 ........................................ Staff medical technologist, Yonsei University Hospital, Seoul, Korea 1967 - 1968 ........................................ Medical technologist (ASCP), Muhlenberg Hospital, Plainfield, NJ 1968 - 1970 ........................................ Staff medical technologist, Muhlenberg Hospital, New Jersey 1970 to present .................................Sr. medical technologist, Clinical Chemistry Laboratory, The Ohio State Univ. Hospitals, Columbus, Ohio 1976 - 1978 ........................................ Technical consultant to Glasrock Inc., Fairbum, Georgia 1982 - 1983 ........................................ Assistant Professor, and Director of Department of Med. Technology, The Inchon College of Nursing and Allied Medical Profession. PUBLICATIONS Lee JY.: Effects of EDTA on the activity of leukocyte alkaline phosphatase. J. New Jersey Med. Tech., 17: 26-32,1969. Lee JY, Lott JA, and Sharma HM.: Biochemical changes induced by Maharishi Amrit Kalash (MAK-4) and MA-208 in diet-induced hypercholesterolemic rabbits. In: Free Radicals in Diagnostic Medicine: A Systems Approach to Laboratory Technologies, Clinical Correlations, and Antioxidant therapy. Armstrong D., ed., Plenum Press, New York, NY. FIELDS OF STUDY Major Field: Pathology TABLE OF CONTENTS Page ACKNOWLEDGMENTS..............................................................................ii V ITA .................................................................................................................iii LIST OF TABLES..........................................................................................ix LIST OF FIGURES........................................................................................xi LIST OF PLATE...........................................................................................xiii LIST OF ABBREVIATIONS......................................................................xiv CHAPTER I. INTRODUCTION...................................................................1 I-A. Introduction ...................................................................................1 CHAPTER II. LITERATURE REVIEW II-A. What Are Free Radicals?............................................................ 3 II-B. Production of Free Radicals in the Human Body ....................4 II-C. Target Molecules of Free Radical Attacks ...............................6 a. Lipids .......................................................................................6 b. Protein ................................................................................ 7 c. Nucleic acids ........................................................................... 8 d. Carbohydrates.........................................................................9 H-D. Free Radical-Induced Tissue Damage and Diseases ............10 a. Membrane damage ................................................................10 b. Endothelial damage and vascular effects ...........................11 c. Lipid peroxidation ................................................................11 II-E. Atherosclerosis .......................................................................... 12 a. Structure and metabolism of LDL ......................................13 b. LDL receptors ....................................................................... 14 c. Oxidative modification of LDL ..........................................15 d. Characteristics of oxidized-LDL ........................................16 e. Atherosclerosis caused by ox-LDL ....................................19 f. Prevention of ox-LDL as an anti-atherosclerotic mechanism............................................................................. 21 II-F. Measurement of Free Radicals ................................................ 21 a. Direct methods ......................................................................21 b. Indirect methods ...................................................................21 II-G. Antioxidants .............................................................................. 22 a. Mechanisms and functions of biological antioxidants ........................................................................... 23 b. Synergistic effects of antioxidants ......................................30 c. Natural vitamin E and synthetic vitamin E ........................31 II-H. Research on M AK ....................................................................32 a. Antioxidant effects of M A K .............................................. 32 b. Antiatherogenic effects of MAK ........................................34 c. Other miscellaneous activities of MAK .............................35 1. Anticarcinogenic activities of MAK ........................35 2. Immunomodulatory effects of M AK ...................... 36 3. Effects of MAK on CNS ........................................... 38 4. Effects of MAK on other conditions ...................... 39 II-I. WHHL Rabbits and Lipid Research ....................................... 39 a. WHHL rabbits as the experimental m odel........................39 b. Probucol as a primary drug of choice in antioxidant research .............................................................40 c. WHHL rabbits used for other experiments ....................... 42 II-J. Problem Statement ................................................................... 43 E-K. Hypothesis ............................................................................... 44 vi CHAPTER ID. THE ANTIOXIDANT AND ANTIATHEROGENIC EFFECTS OF MAK-4 IN WHHL RABBITS...............................46 ni-A. Introduction ............................................................................46 III-B. Materials and Methods ........................................................ 49 a. Chemicals and instruments ................................................49 b. Animals and diets ...............................................................49 c. Blood collection ..................................................................50 d. Maharishi Amrit Kalash-4 (MAK-4)................................51 e. Ingredients and chemical constituents of MAK-4 ..........51 f. Quality control of MAK-4 ................................................