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C Pieters Orcid.Org/ 0000-0002-9025-6960 Evaluating the neuropsychiatric properties of efavirenz in an inflammatory model of schizophrenia C Pieters orcid.org/ 0000-0002-9025-6960 Dissertation submitted in fulfilment of the requirements for the degree Master of Science in Pharmacology at the North West University Supervisor: Dr M Möller-Wolmarans Examination: November 2019 Student number: 25056522 Vir my ewige held: Dadda Kobus Pieters 18 Julie 1972 – 9 Oktober 2014 God of creation There at the start Before the beginning of time With no point of reference You spoke to the dark And fleshed out the wonder of light And as You speak A hundred billion galaxies are born In the vapor of Your breath the planets form If the stars were made to worship so will I I can see Your heart in everything You've made Every burning star A signal fire of grace If creation sings Your praises so will I God of Your promise You don't speak in vain No syllable empty or void For once You have spoken All nature and science Follow the sound of Your voice And as You speak A hundred billion creatures catch Your breath Evolving in pursuit of what You said If it all reveals Your nature so will I I can see Your heart in everything You say Every painted sky A canvas of Your grace If creation still obeys You so will I If the stars were made to worship so will I If the mountains bow in reverence so will I If the oceans roar Your greatness so will I For if everything exists to lift You high so will I If the wind goes where You send it so will I If the rocks cry out in silence so will I If the sum of all our praises still falls shy Then we'll sing again a hundred billion times God of salvation You chased down my heart Through all of my failure and pride On a hill You created The light of the world Abandoned in darkness to die And as You speak A hundred billion failures disappear Where You lost Your life so I could find it here If You left the grave behind You so will I I can see Your heart in everything You've done Every part designed in a work of art called love If You gladly chose surrender so will I I can see Your heart Eight billion different ways Every precious one A child You died to save If You gave Your life to love them so will I Like You would again a hundred billion times But what measure could amount to Your desire You're the One who never leaves the one behind So will I – Hillsong United TABLE OF CONTENTS ACKNOWLEDGMENTS i ABSTRACT iv LIST OF FIGURES vi LIST OF TABLES xiii LIST OF ABBREVIATIONS xv GLOSSARY xxiii CHAPTER 1 1 INTRODUCTION 1 1.1 Dissertation approach and layout 1 1.2 Problem statement 2 1.3 Study questions 5 1.4 Study objectives 6 1.5 Hypothesis 7 1.6 Project layout 7 1.7 Expected outcomes 14 1.8 Ethical considerations 15 References 17 CHAPTER 2 26 LITERATURE REVIEW 26 Human immunodeficiency virus, Acquired Immune Deficiency 1.1 26 Syndrome & Highly active anti-retroviral therapy. 1.2 Efavirenz 27 Efavirenz abuse and mechanism of efavirenz-related neuropsychiatric 1.2.1 28 effects 1.3 Substance-induced psychotic disorder, schizophrenia and addiction 31 1.3.1 Substance-induced psychotic disorder 31 1.3.1.1 Schizophrenia 31 1.3.2 Drug abuse and addiction 33 1.4 Neuroanatomy and neurochemistry 33 1.4.1 Dopamine 34 1.4.1.1 Dopamine transporter 37 Dopamine-and- cyclic adenosine monophosphate -regulated 1.4.1.2 38 phosphoprotein (with a molecular weight of 32kD) 1.4.2 Serotonin 40 1.5 Neurodevelopmental hypothesis 42 1.6 Inflammation 43 1.7 The role of the cerebellum in schizophrenia and addiction 46 1.8 Neuroplasticity 47 1.8.1 c-Fos 48 1.9 Hypothalamic-pituitary-adrenal axis 51 1.10 Oxidative stress 52 1.10.1 N-acetylcysteine 54 1.11 Animal models 57 1.11.1 Maternal immune activation 60 1.11.2 Behavioural analysis 61 Synopsis 63 References 64 CHAPTER 3 107 MANUSCRIPT 107 1. Introduction 111 2. Materials and methods 114 2.1 Statement on ethics 114 2.2 Animals 114 2.3 Study design 114 2.4 Drugs and drug exposure protocol 117 2.5 Body weight 118 2.6 Behavioural analysis 118 2.7 Peripheral and neurochemical analysis 120 2.8 Statistical analysis 122 3. Results 123 3.1 Body weight 123 3.2 Behavioural analysis 123 3.3 Peripheral analysis 130 3.4 Neurochemical analysis 132 4. Discussion 138 References 147 CHAPTER 4 164 CONCLUSION 164 1.1 Concluding remarks 164 1.2 Study outcomes 173 1.3 Limitations and future recommendations 175 References 176 ADDENDUM A 180 ETHICS APPROVAL LETTER 180 ADDENDUM B 182 PRE-NATAL AND POST-NATAL EXPERIMENTAL PROTOCOLS 182 PRE-NATAL PROTOCOL 182 Saline and lipopolysaccharide administration 182 POST-NATAL PROTOCOL 187 Conditioned place preference 188 Locomotor activity 191 Pre-pulse inhibition 192 References 213 ADDENDUM C 216 PERIPHERAL- AND NEUROCHEMICAL ANALYSIS 216 PERIPHERAL ANALYSIS 220 Corticosterone 220 Glutathione 226 NEUROCHEMICAL ANALYSIS 231 c-Fos 232 Dopamine transporters 237 Phosphoprotein phosphatase-1 regulatory subunit 247 1B References 257 ADDENDUM D 259 AUTHOR GUIDELINES 259 Frontiers in Psychiatry 259 ADDENDUM E 279 LETTERS OF CONSENT TO SUBMIT MANUSCRIPT 280 ACKNOWLEDGEMENTS/BEDANKINGS Abba Vader Alle eer kom U toe! Dankie dat U met elke teleurstelling my vasgehou het en met elke klein oorwinning saam fees gevier het! Dankie vir die krag en motivering wat Vader my elke dag gegee het. Mag alles wat ek aanpak altyd tot die eer en verheerliking van U wees. Sanette Pieters Liefste mamma, ek is amper doodseker die dag toe God besluit het om die vrou te skape was mamma in Sy gedagtes. Mamma is alles wat ‘n vrou moet wees; Godvresend, waardig, beeldskoon, intelligent, hardwerkend, sterk (ongelooflik sterk), eerlik en ‘n totale refleksie van God se liefde en lig! Ek sal in my lewe nooit genoeg dankie kan sê vir al die opofferings wat mamma elke liewe dag maak vir my en boetie nie. Alles wat ek is, is tedanke aan mamma. Dr Marisa Möller-Wolmarans Dr Marisa, baie dankie vir die ongelooflike groot leerskool wat dr vir my oor die twee jaar gebied het. Dankie dat dr se deur altyd oopgestaan het en dat daar altyd vir my tyd was. Dankie vir al dr se insette, moeite, geduld en dat ek altyd op dr kon staatmaak. Prof Brian Harvey I will never have the words to explain my admiration for you as a person and researcher. What a privilege to study and work underneath a man of your stature! Prof, thank you so much for all your contributions towards this project and thank you for always being kind and making time despite busy circumstances. Your love and determination in research are inspiring! Prof Linda Brand Prof Brand, baie dankie vir die rol wat Prof elke dag in my lewe gespeel het. Prof het ‘n ongelooflike passie en liefde vir farmakologie wat ek al bewonder vanaf my voorgraadse studies! Dr Stephan Steyn Dr Stephan, sonder dr sou ek seker nogsteeds nie statistiek of die terme daarvan verstaan het nie! Ek sal nooit vir dr genoeg kan dankie sê vir al dr se hulp en insette nie. Niks was ooit te veel moeite vir dr nie! Baie dankie! i Dr De Wet Wolmarans Dr De Wet, dit is ‘n besonderse voorreg om iemand soos dr te ken! Baie dankie vir al dr se raad en ondersteuning deur my meesters, ek het dit so baie waardeer. Dr Francois Viljoen, Dr Malie Rheeders, Dr Makhotso Lekhooa, Prof Tiaan Brink, Prof Douglas Oliver & Cynthia Dabhelia Baie dankie vir elke dag se hope vriendelikheid wat julle na almal uitstraal. Antoinette Fick, Kobus Venter & Walter Dreyer Tannie Antoinette & Kobus – Baie dankie dat ek tydens my eksperimente in die Vivarium elke slag op julle kon staat maak. Julle bystand en raad het vir my baie beteken. Walter, baie dankie vir al jou hulp en bystand met die ELISA kits. Jy is ‘n ongelooflik in jou werk! Willem Cloete Jy verdien die grootste medalje van almal! Dankie dat jy met my uitgehou het tydens my meesters, ek weet ek was soms verskriklik moeilik! Dankie dat jy my altyd net ondersteun en aanmoedig om al my drome uit te leef. Jy is ‘n man na aan God se hart en viraltyd my grootste liefde! Jason Pieters Boetie, jou bydrae tot my meesters is soveel meer as wat jy ooit sal weet! Dankie dat jy elke keer wat ek my journal clubs & colloquim ge-oefen het geduldig geluister het, al het jy nie verstaan waarvan ek praat nie. Dankie dat jy net so ongelooflike boetie is! Annika Anna Britz Jy is ‘n vriendin reguit vanaf God vir my gestuur! Baie dankie dat ek elke liewe goeie en slegte dingetjie met jou kan deel en dat jy altyd saam my bly is en my konstant ondersteun. Dankie dat jy altyd net laatweet “Moet nie worry nie my tjoms, ek bid saam”. Jy is altyd daar maak nie saak wat nie! Willem & Elize Cloete Oom Willem en tannie Elize, baie dankie dat julle huis altyd vir my oop was en dat daar altyd tyd was vir lag en gesels! Dankie vir al julle belangstelling, motivering, liefde en ondersteuning gedurende my meesters, dit het vir my baie beteken. Ek is baie lief vir julle.
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