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Abstract Wagner, Charles Stephen ABSTRACT WAGNER, CHARLES STEPHEN. Evidence Based Validation of Botanical Interventions from Scotland and Medieval Wales. (Under the direction of Dr. Slavko Komarnytsky and Dr. Jillian De Gezelle.) The British Isles are a group of over 6,000 islands in the North Atlantic Ocean. From the first millennium BC onward, various tribes and invaders created a diverse cultural landscape of Celtic, Roman, and Germanic traditions, transforming the isles from barbarian outpost to world superpower. Within this hybrid landscape emerged a rich healing tradition, with a written record spanning approximately 2,000 years – from the Roman era through the Middle Ages, up until the early 20th century. This dissertation addresses the application of traditional Welsh and Scottish botanical interventions to modern health, and using current laboratory analyses, systematically evaluates specific botanicals and formulas for their potential to improve various human health outcomes. Chapter 1 overviews the history of the Celtic linguistic community within the context of plant use and comparatively examines plant species and target health conditions found in the medieval Welsh herbal Meddygon Myddfai against herbal texts from Greco-Roman, Anglo- Saxon, and continental European sources. Although the majority of the recipes were based on the Mediterranean herbal tradition, they preserved unique herbal preparation signatures and six plants could be attributed to the Celtic (Welsh) herbal tradition. This review provides, for the first time, initial evidence for traces of Celtic framework in the wider Western herbal tradition and warrants further investigations of bioactivity and clinical applications of the described plant leads. Chapter 2 examines the antibacterial potential of 83 of 138 plant species indicated in the medieval Welsh herbal Meddygon Myddfai to treat conditions related to microbial infections. In an ethnobotanical survey of the Isle of Arran, a novel field assay for in situ targeted screening of antimicrobials was employed. 67 of the 83 plants identified in Meddygon Myddfai and an additional 14 of 18 plants from local knowledge showed detectable levels of antimicrobial activity. In a follow-up proof-of-concept study, bioassay-guided fractionation was performed to identify bioactive constituents from two high scoring hits that inhibited Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative) bacterial growth. Sabinene, a natural bicyclic monoterpene from juniper "berries" (Juniperus communis L.) and alliin, a natural sulfoxide from garlic cloves (Allium sativum L.), were isolated and confirmed as primary antibacterial leads. Chapter 3 evaluates the anti-colitic potential of a polyherbal recipe from the medieval Welsh herbal Meddygon Myddfai in the dextran sodium sulfate (DSS) mouse model of colitis. The Myddfai recipe, consisting of Artemisia vulgaris L., Plantago major L., and Lamium album L. boiled in goat’s whey, dramatically alleviated body weight loss, disease activity index, and colon length shortening as compared to the DSS control. Beneficial microbiome modulation was also observed. Differential effects were observed from each individual ingredient suggesting that recipe was “designed” by the physicians that recorded it. In Chapter 4, 242 oat (Avena sativa L.) flour samples were analyzed for free, bound, and in vitro digested bitter compounds (avenanthramides A-C, coumaric, ferulic, and caffeic acids.) The samples consisted of 109 varieties from the AFRI CORE diversity panel, each grown at two different locations. Early modern anthropologists noted the consumption of oats, which was not common in continental Europe, to be associated with the robust health of 17th through 19th century Scottish peasants. The acute anti-hyperglycemic effect of selected whole grain oat flours in the polygenic C57BL/6J mouse model of diet-induced obesity was evaluated and correlated to metabolite composition. Pretreatment with oat cultivars high in avenanthramides showed marked reduction of post prandial glucose rise in mice. Collectively, these findings support historical botanical interventions as effective dietary agents to improve human health. When backed by evidence and proper clinical integration, historical botanical interventions have great potential for preventing chronic disease, managing symptoms, and enhancing patient quality of life. © Copyright 2020 by Charles Stephen Wagner All Rights Reserved Evidence Based Validation of Botanical Interventions from Scotland and Medieval Wales by Charles Stephen Wagner A dissertation submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Plant Biology Raleigh, North Carolina 2020 APPROVED BY: _______________________________ _______________________________ Dr. Slavko Komarnytsky Dr. Jillian De Gezelle Committee Co-Chair Committee Co-Chair _______________________________ _______________________________ Dr. Richard L. Blanton Dr. Xu “Sirius” Li DEDICATION To Matt, for teaching me to look at the Moon. “πάντες ἄνθρωποι τοῦ εἰδέναι ὀρέγονται φύσει.” -Aristotle, Metaphysica ii BIOGRAPHY Charles Wagner was born in Louisville, Kentucky. The child of two educators, Charlie was always fascinated by living things and their histories. A love of ideas, books, maps, and music led him to study philosophy at St. John’s College in Annapolis, Maryland before transferring to North Carolina State University, where he completed his bachelor’s degree in Plant Biology. His Orcadian heritage has inspired much of his work and travel, along with a desperate desire to live a simple life close to nature. iii ACKNOWLEDGMENTS I have seen and done a lot of strange and interesting things in my life. A PhD seemed like some natural extension of that. Through good times and bad I know that love and good “raisins” have been my true North. For that, and foremostly, I would like to thank my parents, Cindy and Steve Wagner. It has been a long road to get here and I could not have done it without your grit and determination to see me live well. You have stuck with me for a long time and there are really no words to express the impact this has had on me. Of course, in this same breath I must also include my sister, Elizabeth. There is no one on Earth that understands me the way you do and has shared my most inner life. You taught me to write, and I consider you my closest friend – through thick and thin. Then there is my brother Matthew. He has been the guiding principle in my life and my family’s. He lived a very difficult and complex life, yet he was always fearless, strong willed, and humorous. I am so grateful, and forever indebted to him. He saved me from the frivolity that so many live in and he taught me that life is so much more than what meets the eye. Rest in peace, dear brother. Next, I must thank my advisors and professors at NC State for fostering me as a young researcher. First, I would like to express my gratitude to Dr. Jillian De Gezelle. You enabled me to pursue my true interests and not once pulled me in another direction that better suited you. My journey into ethnobotany started with your classes and guidance and it was there I first learned of Myddfai. Thank you for sticking with me through my 4 semesters as a TA and for always supporting me thereafter. I am also greatly indebted to Dr. Slavko Komarnytsky, a true friend and brilliant person. You brought me into the lab and helped me learn to navigate so many hurdles in academia and iv life. Your expertise in biology and writing as well as your love of philosophy, history, and ideas has been a joy to engage with. I genuinely wish you the best and I will miss our conversations (and hope they continue) – deep and dark. спасибо большое. In addition, I would also like to thank Dr. Chad Jordan and Dr. Larry Blanton. You both took a huge chance on bringing me into the department of Plant Biology at NC State. Dr. Jordan helped me every step of the way in transferring from St. John’s as an undergrad and was a constant source of advice and encouragement. Dr. Blanton, similarly, lead me into graduate school, served on my committee, and has been there to answer every question imaginable without judgement. I would also like to thank a few others, and then there are even more. Thanks to Mickey Wilson, for teaching me countless techniques in the lab and never hesitating to help out. Thanks to Keith Robertson for showing me the plants of bonnie Scotland. I also couldn’t leave out my dear friend Dillon Ade, for pushing me to pursue a healthier lifestyle. Finally, last but not least, I must thank Katelyn DeZego, a wonderful partner and all- around amazing lady. I know that dealing with me is not so easy, as I am always dreaming of my next big undertaking. Your love and support for these past 4 years has saved me from immense loneliness and has kept me on the right path – changing for the better. You have all been instrumental in my success. I will end by thanking all the people that came before me on those dark Isles. Our world is changing faster than it ever has, yet more than ever I feel solidly rooted. I persist because of this identity. On the cliffs of Westray “I felt the touch of the kings and the breath of the wind… with the ghosts of the men who can never sing again...” And for that, I am forever grateful. v TABLE OF CONTENTS LIST OF TABLES ........................................................................................................................ ix LIST
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