Yeung, Tak Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 181-197). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iv / Acknowledgements --- p.vi / List of Publications --- p.vii / Presentation --- p.vii / List of Abbreviations --- p.viii / Abbreviations in Figures --- p.xiii / Abbreviations in Symbols --- p.xiv / List of Cell Lines Used in this Study --- p.xv / Table of Contents --- p.xvi / List of Figures --- p.xxv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Cellular Redox State and Cancer Biology --- p.1 / Chapter 1.2 --- Reactive Oxygen Species (ROS) --- p.1 / Chapter 1.3 --- Regulation of Cellular Redox State by Intrinsic and Extrinsic Antioxidant Systems --- p.5 / Chapter 1.3.1 --- Intrinsic Antioxidant System --- p.6 / Chapter 1.3.2 --- Extrinsic Antioxidant System --- p.8 / Chapter 1.4 --- Glutathione --- p.9 / Chapter 1.4.1 --- General Information of Glutathione --- p.9 / Chapter 1.4.2 --- Functions of Glutathione --- p.12 / Chapter 1.4.2.1 --- As an Antioxidant and Free Radical Scavenger --- p.12 / Chapter 1.4.2.2 --- As a Detoxifier --- p.13 / Chapter 1.4.2.3 --- As a Regulator of Cell Signaling --- p.14 / Chapter 1.4.3 --- Synthesis of Glutathione --- p.15 / Chapter 1.4.4 --- Catabolism of Glutathione --- p.15 / Chapter 1.4.5 --- Transport and Uptake of Glutathione --- p.16 / Chapter 1.4.6 --- Glutathione in Cancer Biology --- p.18 / Chapter 1.4.6.1 --- "Role of Glutathione in the Regulation of Carcinogenesis, Growth and Apoptosis of Cancer Cells" --- p.18 / Chapter 1.4.6.1.1 --- Role of Glutathione in Carcinogenesis --- p.18 / Chapter 1.4.6.1.2 --- Role of Glutathione in the Growth of Cancer Cells --- p.20 / Chapter 1.4.6.1.3 --- Role of Glutathione in Apoptosis of Cancer Cells --- p.21 / Chapter 1.4.6.2 --- Role of Glutathione in the Regulation of Metastasis --- p.23 / Chapter 1.4.6.3 --- Role of Glutathione in Cancer Resistance and Therapy --- p.24 / Chapter 1.4.6.3.1 --- Role of Glutathione in Cancer Resistance --- p.24 / Chapter 1.4.6.3.2 --- Role of Glutathione in Cancer Therapy --- p.24 / Chapter 1.5 --- Aims of the Present Study --- p.25 / Chapter Chapter 2 --- In Vitro Study of Bαicαlein and Baicalin on Glutathione Depletion --- p.28 / Chapter 2.1 --- Introduction --- p.28 / Chapter 2.1.1 --- Scutellaria bαicαlensis Georgi --- p.28 / Chapter 2.1.1.1 --- General Clinical Applications to Treat or Prevent Diseases --- p.28 / Chapter 2.1.1.2 --- As an Antioxidant and Free Radical Scavenger --- p.29 / Chapter 2.1.1.3 --- Long History for Treatment of Cancers with the Obscure Mechanism --- p.30 / Chapter 2.1.1.4 --- Major Components --- p.31 / Chapter 2.1.2 --- Baicalein and Baicalin --- p.32 / Chapter 2.1.2.1 --- General Clinical Applications to Treat or Prevent Diseases --- p.32 / Chapter 2.1.2.2 --- As an Antioxidant and Free Radical Scavenger --- p.33 / Chapter 2.1.3 --- Hypothesis: Baicalein and Baicalin Induce Cancer Cell Death Via Glutathione Depletion --- p.35 / Chapter 2.2 --- Materials and Methods --- p.36 / Chapter 2.2.1 --- Chemicals --- p.36 / Chapter 2.2.2 --- Buffers and Solutions --- p.36 / Chapter 2.2.3 --- Animals --- p.37 / Chapter 2.2.4 --- Preparation of Rat Brain Microsomes --- p.37 / Chapter 2.2.5 --- Glutathione Depletion Assay In Vitro and Thiol Depletion Assay in Rat Brain Microsomes --- p.38 / Chapter 2.2.6 --- Statistical Analysis --- p.39 / Chapter 2.3 --- Results --- p.40 / Chapter 2.3.1 --- Effects of Baicalein and Baicalin on Sulfhydryl Contents of Glutathione --- p.42 / Chapter 2.3.2 --- Effects of Baicalein and Baicalin on Sulfhydryl Contents of Rat Brain Microsomes --- p.42 / Chapter 2.4 --- Discussion --- p.44 / Chapter Chapter 3 --- Effects of Baicalein and Baicalin on Proliferation of Different Human Cancer and Normal Cells --- p.45 / Chapter 3.1 --- Introduction-Importance of Developing A Novel Compound Inducing Cancer Cells to Cell Death with the Least Side Effects on Normal Cells --- p.45 / Chapter 3.2 --- Materials and Methods --- p.46 / Chapter 3.2.1 --- Instruments --- p.46 / Chapter 3.2.2 --- Chemicals and Cell Culture Reagents --- p.46 / Chapter 3.2.3 --- Buffers --- p.46 / Chapter 3.2.4 --- Cell Lines --- p.47 / Chapter 3.2.5 --- Cell Culture --- p.48 / Chapter 3.2.6 --- Determination of Cell Proliferation by MTT Assay --- p.49 / Chapter 3.3 --- Results --- p.51 / Chapter 3.3.1 --- Anti-Proliferative Effects of Baicalein and Baicalin on Different Cancer Cell Lines --- p.51 / Chapter 3.3.2 --- Effects of Baicalein on Different Normal Cell Lines --- p.56 / Chapter 3.4 --- Discussion --- p.58 / Chapter 3.4.1 --- Anti-Proliferative Effects of Baicalein and Baicalin on Different Cancer Cell Lines --- p.58 / Chapter 3.4.2 --- Effects of Baicalein on Cell Proliferation on Different Human Normal Cell Lines --- p.60 / Chapter Chapter 4 --- Glutathione-Depleting Effects of Baicalein on Cell Proliferation of Different Cell Lines --- p.61 / Chapter 4.1 --- Introduction-Brain Tumors --- p.61 / Chapter 4.1.1 --- Types and Classifications of Brain Tumors --- p.61 / Chapter 4.1.2 --- "Incidence Time, Patient Survival Time and Rate for" --- p.65 / Chapter 4.1.3 --- Symptoms and Diagnostic Methods for Brain Tumors --- p.66 / Chapter 4.1.4 --- "Treatments, Side Effects and Difficulties of Treatments for Brain Tumors" --- p.67 / Chapter 4.1.5 --- Glutathione Levels in Brain Normal and Cancer Cells --- p.69 / Chapter 4.2 --- Materials and Methods --- p.70 / Chapter 4.2.1 --- Instruments --- p.70 / Chapter 4.2.2 --- Chemicals --- p.70 / Chapter 4.2.3 --- Buffers --- p.70 / Chapter 4.2.4 --- Determination of Cell Proliferation by MTT Assay --- p.70 / Chapter 4.2.5 --- Determination of Intracellular Glutathione Depletion by Fluorescent Dye CMAC --- p.71 / Chapter 4.2.6 --- Determination of Cellular Reduced Glutathione Levels by DTNB-Coupled Glutathione Reductase Recycling Assay --- p.73 / Chapter 4.3 --- Results --- p.75 / Chapter 4.3.1 --- Effects of Baicalein on Intracellular GSH Levels and Cell Proliferation for Different Cell Lines --- p.75 / Chapter 4.3.2 --- Basal Intracellular Glutathione in Different Cell Lines --- p.81 / Chapter 4.4 --- Discussion --- p.84 / Chapter 4.4.1 --- Intracellular Glutathione Depletion and Cell Death Induction Effects of Baicalein on Different Cell Lines --- p.84 / Chapter 4.4.2 --- Relationship between Basal Glutathione Levels and Drug Susceptibilities --- p.85 / Chapter Chapter 5 --- Effects of Baicalein on Apoptosis and Caspase Pathways --- p.88 / Chapter 5.1 --- Introduction-Modes of Cell Death --- p.88 / Chapter 5.1.1 --- Necrosis --- p.88 / Chapter 5.1.2 --- Apoptosis --- p.89 / Chapter 5.2 --- Materials and Methods --- p.92 / Chapter 5.2.1 --- Chemicals --- p.92 / Chapter 5.2.2 --- Buffers --- p.92 / Chapter 5.2.3 --- Determination of Change of Mitochondrial Membrane Potential by JC-1 --- p.93 / Chapter 5.2.4 --- Determination of Apoptosis by Annexin V-Propidium Iodide Staining --- p.94 / Chapter 5.2.5 --- Determination of Cell Cycle Arrest by Propidium Iodide Staining --- p.95 / Chapter 5.2.6 --- "Determination of Caspase-3, -8 and -9 Activities by Fluorescent-Labeled Peptides" --- p.96 / Chapter 5.2.7 --- Determination of DNA Fragmentation --- p.97 / Chapter 5.2.8 --- Terminal Deoxynucleotidyl Transferase Mediated dUTP End Labeling (TUNEL) Assay --- p.99 / Chapter 5.2.9 --- Flow Cytometry --- p.101 / Chapter 5.3 --- Results --- p.102 / Chapter 5.3.1 --- Effects of Baicalein on Mitochondrial Membrane Potential by JC-1 Staining --- p.102 / Chapter 5.3.2 --- Effects of Baicalein on Apoptosis and Necrosis by Annexin V-Propidium Iodide Staining --- p.104 / Chapter 5.3.3 --- Effects of Baicalein on Cell Cycle Arrest by Propidium Iodide Staining --- p.108 / Chapter 5.3.4 --- "Effects of Baicalein on Caspase-3, -8 and -9 Activities" --- p.110 / Chapter 5.3.5 --- Effeets of Baiealein on DNA Fragmentation --- p.115 / Chapter 5.3.6 --- Effects of Baicalein on TUNEL Assay --- p.117 / Chapter 5.4 --- Discussion --- p.120 / Chapter Chapter 6 --- Pro-Oxidant Role of Baicalein on Reactive Oxygen Species Generation --- p.122 / Chapter 6.1 --- Introduction --- p.122 / Chapter 6.2 --- Materials and Methods --- p.122 / Chapter 6.2.1 --- Chemicals --- p.122 / Chapter 6.2.2 --- Determination of Cellular Reactive Oxygen Species Generation by Fluorescent Dye cDCFDA --- p.123 / Chapter 6.2.3 --- Determination of Mitochondrial Reactive Oxygen Species Generation by Fluorescent Dye Rhl23 --- p.124 / Chapter 6.3 --- Results --- p.125 / Chapter 6.3.1 --- Effects of Baicalein on Cellular ROS Generation by Fluorescent Dye cDCFDA --- p.125 / Chapter 6.3.2 --- Effects of Baicalein on Mitochondrial ROS Generation by Fluorescent Dye Rhl23 --- p.129 / Chapter 6.4 --- Discussion --- p.132 / Chapter Chapter 7 --- The Anticancer Mechanistic Study of Baicalein --- p.133 / Chapter 7.1 --- Introduction --- p.133 / Chapter 7.2 --- Materials and Methods --- p.134 / Chapter 7.2.1 --- Chemicals --- p.134 / Chapter 7.2.2 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death by Different Antioxidant Treatments --- p.134 / Chapter 7.2.3 --- Reversibility of Baicalein-Induced Cellular ROS Generation --- p.136 / Chapter 7.2.4 --- Reversibility of Baicalein-Induced Apoptosis by Co-Treatment of Different Antioxidants and Caspase Inhibitors --- p.137 / Chapter 7.2.5 --- "Reversibility of Baicalein-Induced Caspase-3, -8 and -9 Activation by Co-Treatment of Different Antioxidants" --- p.138 / Chapter 7.3 --- Results --- p.139 / Chapter 7.3.1 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death by Different Antioxidant Treatments --- p.139 / Chapter 7.3.1.1 --- Pre-treatments --- p.139 / Chapter 7.3.1.2 --- Co-treatments --- p.141 / Chapter 7.3.1.3 --- Post-treatments --- p.144 / Chapter 7.3.2 --- Reversibility of Baicalein-Induced Cellular ROS Generation by Co-Treatment of Different Antioxidants --- p.147 / Chapter 7.3.3 --- Reversibility of Baicalein-Induced Apoptosis by Co-Treatment of Different Antioxidants and Caspase Inhibitors --- p.152 / Chapter 7.3.4 --- Reversibility of Baicalein-Induced Caspase-3 Activation by Co-Treatment of Different Antioxidants --- p.156 / Chapter 7.3.5 --- Reversibility of Baicalein-Induced Caspase-8 and -9 Activation by Co-Treatment of Different Antioxidants --- p.160 / Chapter 7.4 --- Discussion --- p.164 / Chapter 7.4.1 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death --- p.164 / Chapter 7.4.2 --- "Reversibility of Baicalein-Induced ROS Generation," --- p.167 / Chapter 7.5 --- Concluding Remarks --- p.168 / Chapter Chapter 8 --- General Discussion --- p.169 / Chapter 8.1 --- Drug Delivery to Brain --- p.169 / Chapter 8.2 --- Protective Roles of Baicalein on Brain Cells --- p.170 / Chapter 8.2.1 --- Actions Against Oxidative Stress --- p.170 / Chapter 8.2.2 --- Actions Against Other Neurotoxic Damages --- p.171 / Chapter 8.2.3 --- Actions Against Neuronal Diseases --- p.172 / Chapter 8.3 --- Anticancer Roles of Baicalein on Astrocytoma --- p.173 / Chapter 8.4 --- Implications on the Dual Roles of Baicalein: Antioxidant and Pro-oxidant --- p.175 / Chapter 8.5 --- Future Perspectives --- p.175 / Chapter 8.5.1 --- Effects of Baicalein on Antioxidant System --- p.175 / Chapter 8.5.2 --- Effects of Baicalein on GSH Synthesis --- p.176 / Chapter 8.5.3 --- In Vivo Studies on Cytotoxic Effects of Baicalein --- p.177 / Chapter 8.5.4 --- In Vivo Studies on Anti-Tumor Effects and In Vitro Studies on Anti-Metastasis Effects of Baicalein --- p.178 / Reference List --- p.181

Zhou, Linli. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 108-118). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Abbreviations --- p.v / List of Figures --- p.vii / List of Tables --- p.ix / Table of Contents --- p.x / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Breast Cancer --- p.1 / Chapter 1.1.1 --- Introduction to Breast Cancer --- p.1 / Chapter 1.1.2 --- Types of Breast Cancer --- p.3 / Chapter 1.1.3 --- Epidemiologic Risk Factors and Etiology --- p.4 / Chapter 1.2 --- Estrogen and Breast Cancer --- p.7 / Chapter 1.3 --- Estrogen Receptor --- p.9 / Chapter 1.4 --- Current Treatment of Breast Cancer --- p.10 / Chapter 1.4.1 --- Chemotherapy --- p.10 / Chapter 1.4.2 --- Hormonal (Anti-Estrogen) Therapy --- p.11 / Chapter 1.4.2.1 --- Tamoxifen and Other Anti-estrogens --- p.12 / Chapter 1.4.2.2 --- Disadvantages of Tamoxifen --- p.13 / Chapter 1.5 --- Arsenic Trioxide --- p.14 / Chapter 1.5.1 --- The Characteristics of Arsenic Trioxide (AS2O3) --- p.14 / Chapter 1.5.2 --- The Medical use of Arsenic Trioxide (As2O3) --- p.16 / Chapter 1.5.3 --- Arsenic Trioxide (As2O3) in treating Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.1 --- Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.2 --- All-trans Retinoic Acid (ATRA) Treatment of APL --- p.18 / Chapter 1.5.3.3 --- Clinical Trial of the Arsenic Trioxide on APL --- p.19 / Chapter 1.5.3.4 --- In vitro and in vivo Study of Arsenic Trioxide (As2O3) in treating APL --- p.19 / Chapter 1.5.3.5 --- Common Side Effects of Arsenic Trioxide (As2O3) on APL --- p.21 / Chapter 1.5.4 --- Anti-cancer effect of Arsenic Trioxide on other cancers --- p.23 / Chapter 1.6 --- Aim of Study --- p.24 / Chapter Chapter 2 --- Materials and Methods --- p.26 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.27 / Chapter 2.1.1.1 --- Cell Lines --- p.27 / Chapter 2.1.1.2 --- Culture Medium --- p.27 / Chapter 2.1.2 --- Chemicals --- p.28 / Chapter 2.1.3 --- Buffers and Reagents --- p.29 / Chapter 2.1.4 --- Reagents for MTT Assay --- p.30 / Chapter 2.1.5 --- Reagents for DNA Fragmentation --- p.31 / Chapter 2.1.5.1 --- Reagents for DNA Extraction --- p.31 / Chapter 2.1.5.2 --- Reagents for Gel Electrophoresis --- p.31 / Chapter 2.1.6 --- Reagents for Western Blotting --- p.32 / Chapter 2.1.6.1 --- Reagents for Protein Extraction --- p.32 / Chapter 2.1.6.2 --- Reagents for SDS-PAGE --- p.33 / Chapter 2.1.7 --- Reagents for Flow Cytometry --- p.36 / Chapter 2.1.8 --- In Vivo Study --- p.37 / Chapter 2.2 --- Methods --- p.38 / Chapter 2.2.1 --- Cell Treatment --- p.38 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.38 / Chapter 2.2.3 --- MTT Assay --- p.38 / Chapter 2.2.4 --- Detection of DNA Fragmentation --- p.39 / Chapter 2.2.5 --- Flow Cytometry --- p.40 / Chapter 2.2.5.1 --- Detection of Cell Cycle Pattern with PI --- p.40 / Chapter 2.2.5.2 --- Detection of Apoptosis with Annexin V-PI --- p.40 / Chapter 2.2.6 --- Western Blot Analysis --- p.41 / Chapter 2.2.6.1 --- Protein Extraction --- p.41 / Chapter 2.2.6.2 --- Protein Concentration Determination --- p.41 / Chapter 2.2.6.3 --- Western Blotting --- p.42 / Chapter 2.2.7 --- In Vivo Study --- p.44 / Chapter 2.2.7.1 --- Animal Model --- p.44 / Chapter 2.2.7.2 --- Treatment Schedule --- p.44 / Chapter 2.2.7.3 --- Toxicity of Arsenic Trioxide --- p.45 / Chapter Chapter 3 --- Anti-Proliferation Effect of As2O3 on MDA-MB-231 cells --- p.47 / Chapter 3.1 --- Study the Anti-proliferation Effect of As2O3 on MDA-MB-231 Cells by MTT Assay --- p.48 / Chapter 3.2 --- Comparsion Anti-proliferation Effect of AS2O3 on MDA-MB-231 Cells to that of Tamoxifen --- p.50 / Chapter 3.3 --- "Study Toxicity of AS2O3 on Normal Breast Cells Line, 184B5" --- p.52 / Chapter 3.4 --- Summary --- p.54 / Chapter Chapter 4 --- Mechanism of Growth Inhibition Effect of As2O3 on MDA-MB-231 cells --- p.56 / Chapter 4.1 --- Cell Cycle Analysis of As2O3 Treated MDA-MB-231 Cells --- p.57 / Chapter 4.2 --- Detection of DNA Fragmentation --- p.60 / Chapter 4.3 --- Detection of Apoptosis Induced by AS2O3 on MDA-MB-231 Cells by Flow Cytometry --- p.62 / Chapter 4.4 --- Regulation of Apoptotic Related Protein by As2O3 on MDA-MB-231 Cells --- p.64 / Chapter 4.4.1 --- Expression Level of Bcl-2 and Bax Protein --- p.66 / Chapter 4.4.2 --- Expression Level of Cytochrome C --- p.69 / Chapter 4.4.3 --- Expression Level of Caspase9 --- p.71 / Chapter 4.4.4 --- Expression Level of FasL --- p.73 / Chapter 4.4.5 --- Expression Level of Caspase8 --- p.75 / Chapter 4.4.6 --- Expression Level of Caspase3 --- p.77 / Chapter 4.4.7 --- Expression Level of Poly (ADP-ribose) Polymerase (PARP) --- p.79 / Chapter 4.4.8 --- Expression Level of p53 --- p.81 / Chapter 4.5 --- Regulation of Cell Cycle Related Protein by AS2O3 on MDA-MB-231 Cells --- p.83 / Chapter 4.5.1 --- Expression Level of Cyclin B --- p.84 / Chapter 4.5.2 --- Expression Level of Cyclin E --- p.86 / Chapter 4.6 --- Summary --- p.88 / Chapter Chapter 5 --- In Vivo Study of Anti-tumor Effect of As2O3 --- p.89 / Chapter 5.1 --- Anti-tumor Effect of AS2O3 on Tumor Bearing Nude Mice --- p.90 / Chapter 5.2 --- Toxic Effect of AS2O3 on Normal Tissues --- p.93 / Chapter 5.3 --- Summary --- p.98 / Chapter Chapter 6 --- Discussion --- p.99 / Chapter 6.1 --- Anti-tumor Effect of AS2O3 on Breast Cancer --- p.100 / Chapter 6.2 --- Induction of Apoptosis and Cell Cycle arrest by AS2O3 --- p.101 / Chapter 6.3 --- Side Effect of AS2O3 on Breast Cancer Treatment --- p.103 / Chapter Chapter 7 --- Future Perspectives --- p.105 / Chapter 7.1 --- Future Perspectives --- p.106 / References --- p.108

Li, Wing Yan Kate. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 130-154). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / Abstract (Chinese Version) --- p.vi / Table of Contents --- p.viii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1 --- General Introduction / Chapter 1.1. --- Overview of cancer --- p.1 / Chapter 1.2. --- Apoptosis and cancer --- p.4 / Chapter 1.3. --- Roles and regulation of caspase-dependent apoptosis --- p.7 / Chapter 1.3.1. --- Extrinsic death receptor pathway --- p.8 / Chapter i. --- TNFR1 and TNFa --- p.13 / Chapter ii. --- CD95/Fas and CD95 Ligand/FasL --- p.14 / Chapter iii. --- "TRAIL-R1(DR4), TRAIL-R2 (DR5) and TRAIL" --- p.14 / Chapter 1.3.2. --- Intrinsic mitochondrial pathway --- p.16 / Chapter i. --- Bcl-2 family of proteins --- p.17 / Chapter ii. --- Reactive Oxygen Species (ROS) --- p.19 / Chapter 1.4. --- Phytochemicals from Traditional Chinese Medicine (TCM) as a source of new therapeutics --- p.22 / Chapter 1.5. --- Biological effects of baicalein --- p.25 / Chapter 1.5.1 --- Roles of baicalein as a lipoxygenase inhibitor --- p.28 / Chapter 1.5.2 --- Dual roles of baicalein as an antioxidant and prooxidant --- p.28 / Chapter 1.5.3 --- "Roles of baicalein as an anti-carcinogenic, anti-proliferative and anti-metastatic agent" --- p.29 / Chapter 1.6. --- Aims of current study --- p.30 / Chapter Chapter 2 --- Effects of Baicalein on Growth and Survival of Human Cancer Cells / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Cell culture --- p.35 / Chapter 2.2.2 --- Measurement of growth and survival of various cell lines --- p.36 / Chapter 2.2.3 --- Statistical analysis --- p.37 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Baicalein retards the growth and survival of human melanoma A375 and colorectal carcinoma Caco-2 --- p.37 / Chapter 2.3.2 --- Baicalein reduces the growth and survival of melanoma A375 but not in normal skin fibroblast Hs68 cells --- p.40 / Chapter 2.4 --- Discussion --- p.42 / Chapter Chapter 3 --- Effects of Baicalein on Cell Cycle and the Apoptosis in Human Melanoma A375 Cells / Chapter 3.1 --- Introduction --- p.44 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Determination of cell cycle changes and quantification of apoptosis --- p.51 / Chapter 3.2.2 --- Immunoblotting --- p.52 / Chapter 3.2.3 --- Inhibition of caspase-8 by caspase-8 inhibitor --- p.54 / Chapter 3.2.4 --- Fluorometric measurement of caspase-3 activity --- p.54 / Chapter 3.2.5 --- Statistical analysis --- p.55 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Baicalein induces S-phase arrest in cell cycle and triggers apoptosis --- p.55 / Chapter 3.3.2 --- Baicalein induces proteolytic inactivation of PARP and activation of caspases --- p.59 / Chapter 3.3.3 --- Caspase-8 is the major initiator caspase eliciting the baicalein-induced apoptosis --- p.62 / Chapter 3.4 --- Discussion --- p.67 / Chapter Chapter 4 --- Effects of Baicalein on the Extrinsic Apoptotic Pathways in Human Melanoma A375 Cells / Chapter 4.1 --- Introduction --- p.72 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Immunoblotting --- p.75 / Chapter 4.2.2 --- Determination of sub-lethal dose of exogenous TRAIL --- p.76 / Chapter 4.2.3 --- Determination of the combinatory effect of exogenous TRAIL and baicalein --- p.76 / Chapter 4.2.4 --- Statistical analysis --- p.77 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Baicalein upregulates the expressions of death receptor 4 (DR4) and death receptor 5 (DR5) --- p.77 / Chapter 4.3.2 --- Baicalein sensitizes the melanoma cells to sub-lethal dose of exogenous TRAIL --- p.80 / Chapter 4.4 --- Discussion --- p.84 / Chapter Chapter 5 --- Effects of Baicalein on the Extrinsic Apoptotic Pathways in Human Melanoma A375 Cells Cancer Cells / Chapter 5.1 --- Introduction --- p.88 / Chapter 5.2 --- Materials and Methods / Chapter 5.2.1 --- Analysis of mitochondrial membrane potential --- p.94 / Chapter 5.2.2 --- Fractionation of cell lysates into cytosolic and mitochondrial fractions for immunoblotting --- p.95 / Chapter 5.2.3 --- Immunoblotting --- p.95 / Chapter 5.2.4 --- Determination of cellular reactive oxygen species (ROS) production --- p.96 / Chapter 5.2.5 --- Verification of ROS generation via the addition of Trolox´ёØ --- p.96 / Chapter 5.2.6 --- Statistical analysis --- p.97 / Chapter 5.3 --- Results / Chapter 5.3.1 --- Baicalein induces mitochondrial membrane depolarization --- p.97 / Chapter 5.3.2 --- Cytochrome c is released in the baicalein-induced mitochondrial membrane depolarization --- p.100 / Chapter 5.3.3 --- Baicalein does not elicit the intrinsic apoptotic pathway via modulation of some better-characterized Bcl-2 family proteins in A375 cells --- p.102 / Chapter 5.3.4 --- Baicalein induces ROS production --- p.105 / Chapter 5.3.5 --- Baicalein induces mitochondrial permeabilization via ROS-mediated mechanisms --- p.108 / Chapter 5.4 --- Discussion --- p.112 / Chapter Chapter 6 --- General Discussion --- p.119 / References --- p.130

Cheung, Kwok Kuen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 226-263). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

The purpose of this thesis is to develop an optimized and effective iterative reconstruction algorithm and hardware acceleration methods that work synonymously together through reconstruction in proton computed tomography, which accurately maps the electron density.

Bibliography: leaves 178-195.

Helicobacter pylori infects more than half of the global population and causes gastric disorders. The increasing development of antibiotic resistance by the bacterium continues to limit treatment options. The identification and characterisation of novel therapeutic targets are necessary for successful future treatment of the infection. One potential target for therapeutic intervention is the gpt gene encoded by hp0735 (jhp0672) in H. pylori strain 26695 (J99). This gene produces a putative xanthine-guanine phosphoribosyltransferase (XGPRTase), an enzyme of the purine salvage synthesis pathway. This project employed theoretical, molecular and biochemical approaches to investigate features of H. pylori gpt and XGPRTase that will serve to ascertain their therapeutic potential. The production of a functional XGPRTase by H. pylori was investigated in cell-free extracts, and the kinetic parameters of this activity were compared to those of purified rXGPRTase enzyme. The three 6-oxopurine substrates were recognised by rXGPRTase and allosteric kinetics were observed for some substrates of the enzyme in cell-free extracts and for purified enzyme. These observations indicate complex regulation and an influence of cellular interactions on activity. Bioinformatics were employed to analyse XGPRTase phylogeny, and threading techniques used to build a structural model of XGPRTase. The enzyme is significantly divergent from the equivalent mammalian enzyme, and modelling identified specific features of the enzyme. Molecular approaches were utilised to analyse the essential role of gpt in H. pylori survival. These included insertional inactivation of the gpt in wild-type H. pylori strains and in mutants possessing a complementing copy of the gene present at the rdxA locus. No mutants were recovered with inactivated gpt possibly as a result of pleiotropic effects. Plasmid-mediated complementation was attempted employing IPTG-inducible shuttle vectors and did not yield any mutants. Further characterisation of H. pylori XGPRTase was performed by determining the effects of nucleotide monophosphates and purine analogues on enzyme activity. Inhibition by GMP was observed in all cases, however differences in the inhibition by other nucleotide monophosphates were found between cell-free extracts and the recombinant enzyme. Inhibition of rXGPRTase activity was observed by the purine analogue 6-mercaptopurine ribose, a compound that previously has been shown to inhibit H. pylori growth in culture.

Bibliography: leaves 178-195. / xiv, 195 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Chemistry, 2001

"January 2004" / Bibliography: leaves 163-184. / v, 184 leaves : ill. (some col.), photos ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, School of Chemistry and Physics, Discipline of Chemistry, 2004

Graduation date: 1992