by Josephine Wing Sze Lau. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 84-112). / Abstracts in English and Chinese. / Acknowledgment --- p.iii / Abstract --- p.iv / List of Abbreviations --- p.viii / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- Methamphetamine (METH) --- p.1 / Chapter 1.1.1 --- Historical Background and Epidemiology --- p.1 / Chapter 1.1.2 --- Physical Effects of METH --- p.4 / Chapter 1.1.3 --- Neurochemical Alternation of METH --- p.6 / Chapter 1.2 --- Mechanisms of METH Toxicity / Chapter 1.2.1 --- Oxidative Stress --- p.8 / Chapter 1.2.1.1 --- Superoxide (O2-) and Superoxide Dismutase (SOD) --- p.10 / Chapter 1.2.1.2 --- "Hydrogen Peroxide (H202), Catalase and Glutathione (GSH)" --- p.11 / Chapter 1.2.1.3 --- Hydroxyl Radical (OH.) --- p.12 / Chapter 1.2.1.4 --- Nitric Oxide (NO) --- p.13 / Chapter 1.2.2 --- Apoptosis --- p.16 / Chapter 1.2.3 --- Excitotoxicity --- p.17 / Chapter 1.2.4 --- Mitochondrial Dysfunction --- p.18 / Chapter 1.2.5 --- Hyperthermia --- p.21 / Chapter 1.2.5.1 --- Cyclooxygenase-2 (COX-2) --- p.23 / Chapter 1.2.5.2 --- Heme-oxygenase-1 (HO-1) --- p.25 / Chapter 1.2.5.3 --- The Effects of Nitric Oxide (NO) on COX-2 and HO-1 Expressions --- p.27 / Chapter 1.3 --- Astrocytes / Chapter 1.3.1 --- Characteristics of Astrocytes --- p.29 / Chapter 1.3.2 --- Astrocyte Functions --- p.30 / Chapter 1.3.3 --- The Role of Astrocytes in METH-induced Neurotoxicity --- p.34 / Chapter 1.4 --- Aim of Project --- p.37 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS / Chapter 2.1 --- Cell Cultures / Chapter 2.1.1 --- Astrocyte Cultures --- p.42 / Chapter 2.1.2 --- CATH.a Cell line and Astrocytes Co-cultures --- p.43 / Chapter 2.2 --- Treatment / Chapter 2.2.1 --- METH Treatment --- p.44 / Chapter 2.2.2 --- Inhibition of Cyclooxygenase-2 (COX-2) and Inducible Nitric Oxide Synthase (iNOS) --- p.44 / Chapter 2.3 --- Lactate Dehydrogenase (LDH) Assay --- p.45 / Chapter 2.4 --- Assay for Reactive Oxygen Species (ROS) Formation --- p.47 / Chapter 2.5 --- Assay for Adenosine Triphosphate (ATP) Content --- p.48 / Chapter 2.6 --- Determination of Mitochondrial Membrane Potential (Δ Ψm) --- p.50 / Chapter 2.7 --- Determination of Nitrite Levels in Cultured Astrocytes --- p.51 / Chapter 2.8 --- Western Blot Analysis --- p.52 / Chapter 2.8.1 --- COX-2 --- p.53 / Chapter 2.8.2 --- HO-1 --- p.53 / Chapter 2.9 --- Viability Assay of CATH.a-Astrocyte Cocultures --- p.54 / Chapter 2.10 --- Statistics --- p.55 / Chapter CHAPTER THREE: --- RESULTS / Chapter 3.1 --- The Effects of METH Treatment on Cultured Astrocytes / Chapter 3.1.1 --- Lactate Dehydrogenase (LDH) Activities --- p.56 / Chapter 3.1.2 --- Morphological Changes --- p.56 / Chapter 3.1.3 --- The Production of Reactive Oxygen Species / Chapter 3.1.3.1 --- Rate of change (0-120 min) --- p.57 / Chapter 3.1.3.2 --- Time course (0 - 48 h) --- p.57 / Chapter 3.1.4 --- Change in ATP Content --- p.58 / Chapter 3.1.5 --- Change in Mitochondrial Membrane Potential (Δ Ψm) --- p.59 / Chapter 3.1.6 --- Nitrite levels after METH treatment / Chapter a) --- Striatal astrocytes --- p.59 / Chapter b) --- Mesencephalic astrocytes --- p.60 / Chapter c) --- Cortical astrocytes --- p.60 / Chapter 3.1.7 --- The Effects of Aminoguanidine (AG) on Nitrite Levels / Chapter a) --- Striatal astrocytes --- p.61 / Chapter b) --- Mesencephalic astrocytes --- p.62 / Chapter c) --- Cortical astrocytes --- p.62 / Chapter 3.1.8 --- The Effects of Indomethacin (INDO) on Nitrite Levels / Chapter a) --- Striatal astrocytes --- p.63 / Chapter b) --- Mesencephalic astrocytes --- p.64 / Chapter c) --- Cortical astrocytes --- p.64 / Chapter 3.1.9 --- Change in Cyclooxygenase-2 (COX-2) Protein Levels / Chapter a) --- Striatal astrocytes --- p.65 / Chapter b) --- Mesencephalic astrocytes --- p.65 / Chapter c) --- Cortical astrocytes --- p.66 / Chapter 3.1.10 --- Change in Heme-oxygenase-1 (HO-1) Protein Levels / Chapter a) --- Striatal astrocytes --- p.66 / Chapter b) --- Mesencephalic astrocytes --- p.66 / Chapter c) --- Cortical astrocytes --- p.67 / Chapter 3.2 --- Cell Viability on CATH.a-Astrocyte Cocultures After METH Treatment --- p.67 / Chapter CHAPTER FOUR: --- DISCUSSION AND CONCLUSION --- p.69 / REFERENCES --- p.84

: Methamphetamine-related deaths have been rising along with those involving synthetic opioids, mostly fentanyl and fentanyl analogs (FAs). However, the extent to which methamphetamine involvement in deaths differs from those changes occurring in synthetic opioid involvement is unknown.: To determine the patterns and temporal changes in methamphetamine-related deaths with and without other drug involvement.: Data from all methamphetamine-related deaths in West Virginia from 2013 to 2018 were analyzed. Quasi-Poisson regression analyses over time were conducted to compare the rates of change in death counts among methamphetamine and fentanyl//FA subgroups.: A total of 815 methamphetamine-related deaths were analyzed; 572 (70.2%) were male and 527 (64.7%) involved an opioid. The proportion of methamphetamine only deaths stayed relatively flat over time although the actual numbers of deaths increased. Combined fentanyl/FAs and methamphetamine were involved in 337 deaths (41.3%) and constituted the largest increase from 2013 to 2018. The modeling of monthly death counts in 2017-2018 found that the average number of deaths involving fentanyl without methamphetamine significantly declined (rate of change -0.025, < .001), while concomitant fentanyl with methamphetamine and methamphetamine only death counts increased significantly (rate of change 0.056 and 0.057, respectively, < .001).: Fentanyl and FAs played an increasingly significant role in methamphetamine-related deaths. The accelerating number of deaths involving fentanyl/FAs and methamphetamine indicates the importance of stimulants and opioids in unintentional deaths. Comprehensive surveillance efforts should continue to track substance use patterns to ensure that appropriate prevention programs are undertaken.