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ANTIOXIDANT POTENTIAL of SPECIFIC CEREAL GRAIN FRACTIONS: in VITRO and in VIVO STUDIES by KABO MASISI a Thesis Submitted To

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ANTIOXIDANT POTENTIAL of SPECIFIC CEREAL GRAIN FRACTIONS: in VITRO and in VIVO STUDIES by KABO MASISI a Thesis Submitted To ANTIOXIDANT POTENTIAL OF SPECIFIC CEREAL GRAIN FRACTIONS: IN VITRO AND IN VIVO STUDIES By KABO MASISI A thesis Submitted to the Faculty of Graduate Studies of The University of Manitoba In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Department of Food Sciences, Faculty of Agricultural and Food Science University of Manitoba Winnipeg, MB, CANADA Copyright © 2016 by Kabo Masisi ACKNOWLEDGMENT It is with gratitude to acknowledge individuals for their valuable time and support and, organizations that provided funding and support for my research and program. First and foremost, I wish to acknowledge the financial support rendered by Dr. Trust Beta’s research program supported by Canada Research Chairs program and Natural Sciences and Engineering Research Council of Canada (NSERC) discovery grant; International Graduate Student Scholarship (IGSS), Graduate Enhancement of Tri-Council Stipends Grant (GETS) and BMO Financial Group Research Scholarships for Excellence which enabled me to read for a doctoral degree. This has been a most rewarding experience both inside and outside the classroom and research lab. I am also thankful to the Department of Food Science (Laboratory of Food Carbohydrates), Richardson Centre for Functional Foods and Nutraceuticals, Canadian Centre for Agri-Food Research in Health and Medicine (CCARM), Children’s Hospital Research Institute of Manitoba (CHRIM) (formerly Manitoba Institute of Child Health; MICH) and St-Boniface Hospital Albrechtsen Research Centre for providing the great facilities to conduct this research. I wish to express my profound appreciation to my supervisor Professor Trust Beta for giving me the opportunity to work on this very interesting project, painstakingly reading the drafts of this work and providing me with exceptionally insightful comments; and for her mentorship and leadership. Thank you for always encouraging me to consider other possibilities and for being patient as I tried new things. A special thank you also goes to my co-supervisor Professor Mohammed H. Moghadasian for the pathology training, painstakingly reading the drafts of this work (manuscripts) and providing me with exceptionally insightful comments; and for his mentorship and leadership. Thank you for I always encouraging me to consider other possibilities and for being patient as I tried new things. Special thanks also goes to Professor William L. Diehl-Jones for the molecular biology, cell biology and physiology training, painstakingly reading the draft of this work (manuscript) and providing me with exceptionally insightful comments and Professor Martin Scanlon for painstakingly reading the drafts of this work and providing me with exceptionally insightful comments. I wish to express my profound appreciation to Dr. Joseph Gordon for always sharing exceptionally insightful information with me. My sincerest gratitude also goes to the faculty, support staff and students of the Department of Food Science, CCARM, CHRIM and St-Boniface Hospital Albrechtsen Research Centre for creating a supportive learning environment. Special thank you to Alison Ser, Yang Qiu, Pat Heritage, Donald Chapman, Dennis Labossiere, Khuong Le and Dr. Babak Sobhi for technical support and my lab group members (Dr. Victoria Uchizi Ndolo, Dr. Lovemore Nkhata Malunga, Kuuku Biney, Lilei Yu, Yuwei Song, Nora Ghazzawi, Wajihah Mughal, Adewale Michael Esan and Dr. Rokiatou Kone-Berethe). My thanks also go to summer students (Nha Ky Huynh and Tamara Paestch). I would also like to thank the R.O. Burrell staff for their assistant with care of the mice used in this study. I also wish to thank my father Tumediso Masisi, my brother Kanokang Masisi, my sister Segakolodi Masisi and my cousin Lesego Gabaitire for their love and support and to apologize to them for being an absent son and brother. Last but not least to all my relatives and friends in Botswana, Canada, Japan, South Africa and other parts of the world for all the support, solidarity and friendship they have shown me as I staggered and faltered through my uncertain journey towards completion of this work. II POST DEFENSE ACKNOWLEDGEMENT I would like to extend my appreciation to the external examiner Professor David Kitts for being available at the time of my defense and for painstakingly reading my thesis and providing me with exceptionally insightful comments. III DEDICATION To the pillars of my life: God, my parents and grandmother. Without you, my life would fall apart. I might not know where the life’s road will take me, but walking with you, God, through this journey has given me strength. To the loving memory of my grandmother Natefho Mmolai (1924-2005), for sowing in me the seed of discipline, self respect, passion for better things in life. To the loving memory of my mother Gabokgwewi M. Masisi (1960-2006), for sowing in me the seed of discipline, self respect, passion for better things in life. Daddy (Tumediso Masisi), you always told me to “reach for the stars.” I think I got one. Thanks for inspiring my life for transformation. We made it… IV DECLARATION I hereby declare that I KABO MASISI is the sole author of this thesis and have not been accepted in any previous application for any degree. This is a true copy of the original thesis. It is a true record of work done by myself, unless otherwise stated. All sources of information have been duly acknowledged by means of references and copyright licenses for previously published materials have been obtained. The research was conducted in compliance with the ethical standards and guidelines of the University of Manitoba. V THESIS FORMAT This thesis has been prepared in the manuscript format. Referencing style used is for the Food Chemistry Journal. The thesis has seven chapters of which Chapters 1 and 2 are overall introduction and literature review. The experimental chapters are divided in three sections, Section A (Biochemical Model: Chapter 3), Section B (In vitro Model: Chapter 4) and Section C (In vivo Model: Chapter 5 and Chapter 6). Chapter 7 provides the general discussion, conclusions and areas for further research. Experimental Chapters Section A: Biochemical Model Chapter 3: Characterization, identification, quantification and antioxidant capacity of carotenoids of aleurone, germ and endosperm fractions of barley, corn and wheat Section B: In vitro Model Chapter 4: Carotenoids of aleurone, germ, and endosperm fractions of barley, corn and wheat differentially inhibit oxidative stress. Section C: In vivo Model Chapter 5: Dietary corn fractions reduce atherogenesis in LDL receptor knockout mice Chapter 6: Dietary corn fractions attenuate atherosclerosis in LDL receptor knockout mice by differentially modifying antioxidant enzymes activities and oxidative indicators VI TABLE OF CONTENTS ACKNOWLEDGMENT ......................................................................................................... I POST DEFENSE ACKNOWLEDGEMENT ...................................................................... III DEDICATION ..................................................................................................................... IV DECLARATION ................................................................................................................... V THESIS FORMAT ............................................................................................................... VI TABLE OF CONTENTS ................................................................................................... VII LIST OF TABLES ............................................................................................................ XIII LIST OF FIGURES ........................................................................................................... XIV LIST OF ABBREVIATIONS ........................................................................................ XVIII ABSTRACT ...................................................................................................................... XXI CHAPTER 1 ........................................................................................................................... 1 Introduction, Rationale, Hypotheses and Objectives.............................................................. 1 1.1 Introduction ................................................................................................................... 1 1.2 Rationale ....................................................................................................................... 4 1.3 Hypotheses and Objectives ........................................................................................... 4 References .............................................................................................................................. 8 CHAPTER 2 ......................................................................................................................... 13 Literature Review ................................................................................................................. 13 2.1 Oxidative Stress .......................................................................................................... 13 2.1.1 Reactive Oxygen Species ..................................................................................... 14 2.1.2 Antioxidants Defenses.......................................................................................... 18 2.2 Oxidative Stress on Cellular Components .................................................................
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