by Chan Po-cheung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 218-244). / Acknowledgement --- p.i / Abbreviations --- p.ii / Abstract --- p.iv / Table of content --- p.ix / Chapter Chapter 1. --- Biochemistry of Tumor Necrosis Factor --- p.1 / Chapter I. --- Introduction --- p.1 / Chapter 1.1 --- The discovery of tumor necrosis factor (TNF) --- p.1 / Chapter 1.2 --- TNF as an antitumor agent --- p.3 / Chapter 1.3 --- Production of TNF --- p.4 / Chapter 1.4 --- Structure of TNF --- p.5 / Chapter 1.5 --- TNF receptor --- p.6 / Chapter 1.6 --- Biological activities of TNF --- p.10 / Chapter 1.7 --- Anti-tumor activity of TNF --- p.14 / Chapter 1.7.1 --- In vitro studies --- p.14 / Chapter 1.7.1.1 --- Synergistic effect of other cytokines --- p.14 / Chapter 1.7.1.2 --- DNA damages --- p.15 / Chapter 1.7.1.3 --- Free radical generation --- p.15 / Chapter 1.7.1.4 --- Utilization of ATP --- p.16 / Chapter 1.7.1.5 --- Phospholipase A2 activation --- p.17 / Chapter 1.7.2 --- In vivo studies --- p.17 / Chapter 1.8 --- Clinical trials --- p.18 / Chapter Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Materials --- p.20 / Chapter 2.2 --- Solutions commonly used --- p.21 / Chapter 2.3 --- Methods and procedure --- p.23 / Chapter 2.3.1 --- Culture of L-929 cells --- p.23 / Chapter 2.3.2 --- Trypan Blue exclusion test --- p.23 / Chapter 2.3.3 --- Determination of viability of L-929 cells upon rhTNF treatment --- p.24 / Chapter 2.3.4 --- Determination of cellular cAMP level --- p.25 / Chapter 2.3.5 --- Determination of inositol phosphate turnover --- p.26 / Chapter 2.3.6 --- Use of fluorescence probe in the study of rhTNF mediated killing --- p.28 / Chapter 2.3.6.1 --- Determination of changes in internal pH of L-929 cells --- p.29 / Chapter 2.3.6.2 --- Determination of intracellular calcium level in L-929 cells --- p.30 / Chapter 2.3.6.3 --- Determination of membrane potential by fluorescence probes --- p.32 / Chapter 2.3.6.4 --- "Translocation of nucleolar protein, nucleophosmin (B23)in L-929 cells" --- p.32 / Chapter 2.3.6.5 --- Determination of calcium mobilization in L-929 cells by confocal microscopy --- p.34 / Chapter 2.3.6.6 --- Determination of protein kinase C and phospho-tyrosine kinase in L-929 cells --- p.34 / Chapter 2.3.7 --- Uptake of 45Ca2+ in L-929 cells --- p.35 / Chapter 2.3.8 --- Measurement of membrane potential by Patch-clamp assay --- p.36 / Chapter 2.3.9 --- Determination of tyrosine kinase activation by Western blotting --- p.36 / Chapter 2.3.10 --- Statistical analysis --- p.38 / Chapter Chanter 3. --- Effect of rhTNF treatment on nucleophosmin in L-929 cells --- p.39 / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Results --- p.43 / Chapter 3.2.1 --- Effect of TNF (in the presence or absence of actinomycin D) on the nucleophosmin translocation in L-929 cells --- p.43 / Chapter 3.2.2 --- Effect of actinomycin D on the TNF-mediated cytotoxicity on L-929 cells --- p.51 / Chapter 3.3 --- Discussion --- p.57 / Chapter Chapter 4. --- Changes in membrane potential and intracellular pH in rhTNF-mediated cytotoxicity in L-929 cells --- p.59 / Chapter 4.1 --- Introduction --- p.59 / Chapter 4.2 --- Results --- p.61 / Chapter 4.2.1 --- Effect of rhTNF on the membrane potential of L-929 cells determined by fluorescence method --- p.61 / Chapter 4.2.2 --- Effect of rhTNF on the membrane potential of L-929 cells determined by patch clamp technique --- p.64 / Chapter 4.2.3 --- "Effect of K+, Na+ and pH on the rhTNF-mediated cytotoxicity on L-929 cells" --- p.67 / Chapter 4.3 --- Discussion --- p.90 / Chapter Chapter 5. --- Effect of intracellular cAMP and cAMP-dependent protein kinase (PKA) on the rhTNF-mediated cytotoxicity on L-929 cells --- p.92 / Chapter 5.1 --- Introduction --- p.92 / Chapter 5.1.1 --- "GTP-binding protein (G protein), cAMP and protein kinase A" --- p.92 / Chapter 2.1.2 --- Role of cAMP as second messenger --- p.96 / Chapter 5.1.3 --- Bacterial toxin used for study of G-protein --- p.98 / Chapter 5.1.4 --- Effect of cAMP on rhTNF cytotoxicity --- p.99 / Chapter 5.1.5 --- Effect of cAMP-dependent protein kinase (PICA) on rhTNF cytotoxicity --- p.101 / Chapter 5.2 --- Results --- p.102 / Chapter 5.2.1 --- Cyclic-AMP (cAMP) level in rhTNF-treated L-929 cells --- p.102 / Chapter 5.2.2 --- Effect of intracellular cAMP level on rhTNF-mediated cytotoxicity on L-929 cells --- p.104 / Chapter 5.2.3 --- Effect of agonist and inhibitor of cAMP dependent protein kinase (protein kinase A) on rhTNF-mediated cytotoxicity on L-929 cells --- p.107 / Chapter 5.2.4 --- Effect of protein kinase A inhibitors on rhTNF-mediated cytotoxicity on L-929 cells --- p.111 / Chapter 5.3 --- Discussion --- p.118 / Chapter Chapter 6. --- "Role of intracellular free calcium, ions and calcium dependent response in rhTNF-mediated cytotoxicity on L-929 cells" --- p.121 / Chapter 6.1 --- Introduction --- p.121 / Chapter 6.1.1 --- Inositol triphosphate and intracellular free calcium --- p.121 / Chapter 6.1.2 --- Diacylglycerol --- p.131 / Chapter 6.1.3 --- Protein kinase C (PKC) --- p.131 / Chapter 6.1.4 --- Intracellular free calcium ions and protein kinase C --- p.134 / Chapter 6.1.5 --- Effect of intracellular free calcium ions and protein kinase C on TNF-mediated cytotoxicity --- p.135 / Chapter 6.1.6 --- Tyrosine kinase induced release of IP3 --- p.136 / Chapter 6.1.7 --- Calcium channels --- p.136 / Chapter 6.2 --- Result --- p.139 / Chapter 6.2.1 --- Effect of rhTNF on intracellular free [Ca2+] of L-929 cells --- p.141 / Chapter 6.2.2 --- Effect of calcium ion channel blockers on rhTNF-mediated cytotoxicity on L-929 cells --- p.148 / Chapter 6.2.3 --- Effect of protein kinase C (PKC) on rhTNF-mediated cytotoxicity on L-929 cells --- p.158 / Chapter 6.2.4 --- Immunofluorescence staining of PKC in rhTNF-treated L-929 cells --- p.162 / Chapter 6.2.5 --- Effect of calmodulin and calmodulin sensitive calcium ATPase on rhTNF-mediated cytotoxicity on L-929 cells --- p.165 / Chapter 6.2.6 --- Role of inositol triphosphate in rhTNF-mediated cytotoxicity on L-929 cells --- p.167 / Chapter 6.2.7 --- Role of tyrosine kinase activity in the rhTNF-mediated cytotoxicity on L-929 cells --- p.185 / Chapter 6.3 --- Discussion --- p.191 / Chapter Chapter 7. --- Effect of antioxidants on rhTNF-mediated cytotoxicity on L-929 cells --- p.195 / Chapter 7.1 --- Introduction: Oxygen free radicals as mediators of rhTNF-induced tumor cell necrosis --- p.195 / Chapter 7.2 --- Results --- p.199 / Chapter 7.3 --- Discussion --- p.203 / Chapter Chapter 8. --- General Discussion --- p.205 / Bibliography --- p.217

by Joyce Chui Kwan Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 61-75). / Abstracts in English and Chinese. / Acknowledgments --- p.i / List of Abbreviations --- p.iii / Abstract / English --- p.1 / Chinese --- p.2 / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Tumor Formation --- p.3 / Chapter 1.2 --- Anti-tumor Pathways --- p.4 / Chapter 1.3 --- Aim of Project --- p.13 / Chapter Chapter 2 --- The In Vivo effect of Polysaccharopeptide / Chapter 2.1 --- Introduction --- p.15 / Chapter 2.2 --- Materials and Methods --- p.17 / Chapter 2.3 --- Results --- p.18 / Chapter 2.4 --- Discussion --- p.19 / Chapter Chapter 3 --- Cytotoxicity / Chapter 3.1 --- Introduction --- p.23 / Chapter 3.2 --- Materials and Methods --- p.26 / Chapter 3.3 --- Results --- p.28 / Chapter 3.4 --- Discussion --- p.28 / Chapter Chapter 4 --- Anti-angiogenic Effect / Chapter 4.1 --- Introduction --- p.30 / Chapter 4.2 --- Materials and Methods --- p.35 / Chapter 4.3 --- Results --- p.39 / Chapter 4.4 --- Discussion --- p.42 / Chapter Chapter 5 --- Immunomodulation / Chapter 5.1 --- Introduction --- p.45 / Chapter 5.2 --- Materials and Methods --- p.47 / Chapter 5.3 --- Results --- p.50 / Chapter 5.4 --- Discussion --- p.52 / Chapter Chapter 6 --- General Discussion --- p.57 / References --- p.61

by Cheung Yuen-Nei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 194-208). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Contents --- p.vi / List of Figures --- p.xii / List of Tables --- p.xv / Abbreviations --- p.xvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1 --- Introduction --- p.2 / Chapter 1.1 --- Characteristic of Saponins --- p.3 / Chapter 1.1.1 --- Occurrence of Saponins --- p.3 / Chapter 1.1.2 --- General Properties of Saponins --- p.3 / Chapter 1.1.2.1 --- Emulsifying Agents --- p.3 / Chapter 1.2.2.2 --- Forming Complex with Cholesterol --- p.4 / Chapter 1.1.2.3 --- Hemolytic Property --- p.4 / Chapter 1.1.3 --- Structure of Saponins --- p.5 / Chapter 1.1.3.1 --- Categories of Saponins --- p.5 / Chapter 1.1.3.1.1 --- Triterpene Saponins --- p.5 / Chapter 1.1.3.1.2 --- Steroid Saponins --- p.5 / Chapter 1.1.3.2 --- Monodesmosidic and Bidesmosidic Saponins --- p.7 / Chapter 1.1.4 --- Biological and Pharmacological Properties of Saponins --- p.9 / Chapter 1.1.4.1 --- Anti-microbial Activity --- p.9 / Chapter 1.1.4.1.1 --- Anti-fungal Activities --- p.9 / Chapter 1.1.4.1.2 --- Anti-bacterial Activities --- p.10 / Chapter 1.1.4.1.3 --- Anti-viral Activities --- p.10 / Chapter 1.1.4.2 --- Insecticidal Activity --- p.10 / Chapter 1.1.4.3 --- Molluscicidal Activity --- p.10 / Chapter 1.1.4.4 --- Hypocholesterolemic Activity --- p.11 / Chapter 1.1.4.5 --- Anti-ulcer Activity --- p.11 / Chapter 1.1.4.6 --- Contraceptive Activity --- p.12 / Chapter 1.1.4.7 --- Immunomodulatory Activities --- p.12 / Chapter 1.1.4.7.1 --- Direct Immunostimulation --- p.12 / Chapter 1.1.4.7.2 --- Acting as Immuno-adjuvants --- p.13 / Chapter 1.1.4.8 --- Anti-tumor Activity --- p.14 / Chapter 1.1.4.8.1 --- Anti-carcinogenesis --- p.15 / Chapter 1.1.4.8.2 --- Suppression of Tumor Growth --- p.16 / Chapter 1.1.5 --- Anti-tumor Activity of Steroid Saponins --- p.18 / Chapter 1.1.5.1 --- Diosgenin Steroid Saponin --- p.18 / Chapter 1.1.5.2 --- Hong Kong Compounds --- p.18 / Chapter 1.1.5.3 --- Hong Kong18 --- p.21 / Chapter 1.2 --- Human Hepatocellular Carcinoma (HCC) --- p.24 / Chapter 1.2.1 --- The Incidence of Liver Cancer --- p.24 / Chapter 1.2.2 --- Classification of Liver Cancer --- p.24 / Chapter 1.2.3 --- Human Hepatocellular Carcinoma Cell Lines --- p.25 / Chapter 1.2.3.1 --- Human Hepatocellular Carcinoma Cell Line HepG2 --- p.25 / Chapter 1.2.3.2 --- Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.27 / Chapter 1.2.3.2.1 --- Mechanisms of Multidrug Resistance --- p.28 / Chapter 1.2.3.2.2 --- Structure and Characteristics of P-glycoprotein --- p.29 / Chapter 1.2.3.2.3 --- Methods in Dealing with P-glycoprotein Over-expressed MDR Cells --- p.31 / Chapter 1.3 --- Objectives of the Project --- p.32 / Chapter 1.3.1 --- Study of the Anti-tumor Activities of Hong Kong 18 on Human Hepatocellular Carcinoma Cell Line HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter 1.3.2 --- Study of the Anti-tumor Activities of Hong Kong 18on Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter Chapter 2 --- Materials and Methods --- p.33 / Chapter 2.1 --- Materials --- p.34 / Chapter 2.1.1 --- Cell Culture --- p.34 / Chapter 2.1.1.1 --- Cell Lines --- p.34 / Chapter 2.1.1.2 --- Culture Media --- p.35 / Chapter 2.1.2 --- Reagents and Buffers --- p.36 / Chapter 2.1.2.1 --- Phosphate Buffered Saline (PBS) --- p.36 / Chapter 2.1.2.2 --- Reagents and Buffers for DNA Fragmentation --- p.36 / Chapter 2.1.2.3 --- Reagents and Buffers for Western Analysis --- p.37 / Chapter 2.1.2.4 --- Reagents and Buffer for Caspases Activities --- p.39 / Chapter 2.1.2.5 --- Fluorescent Dyes used for Flow Cytometry --- p.39 / Chapter 2.1.3 --- Chemicals --- p.39 / Chapter 2.2 --- Methods --- p.46 / Chapter 2.2.1 --- MTT Assay --- p.46 / Chapter 2.2.2 --- Determination of Cell Viability --- p.46 / Chapter 2.2.3 --- Purification of Macrophages from balb/c Mice --- p.47 / Chapter 2.2.4 --- Hemolysis Assay --- p.47 / Chapter 2.2.5 --- In vivo Studies of the Toxicity of HK18 --- p.48 / Chapter 2.2.6 --- DNA Fragmentation Assay --- p.50 / Chapter 2.2.7 --- Detection of Apoptotic and Necrotic / Late Apoptotic Cells Death by Flow Cytometry with Annexin V-FITC / PI --- p.51 / Chapter 2.2.8 --- Detection of Mitochondrial Membrane Potential by JC-1 Fluorescent Dye --- p.52 / Chapter 2.2.9 --- Detection of Intracellular Ca Level by Flow Cytometry with Fluo-3 Fluorescent Dye --- p.52 / Chapter 2.2.10 --- Detection of Intracellular Hydrogen Peroxide Level by Flow Cytometry with DCF Fluorescence Dye --- p.53 / Chapter 2.2.11 --- Simultaneous Detection of Mitochondrial Membrane Potential and Intracellular Ca2+ or Mitochondrial Membrane Potential and Intracellular Hydrogen Peroxide --- p.54 / Chapter 2.2.12 --- Western Analysis --- p.55 / Chapter 2.2.12.1 --- Total Protein Extraction --- p.55 / Chapter 2.2.12.2 --- Extraction of Cytosolic Proteins --- p.59 / Chapter 2.2.13 --- Determination of Caspases Enzymatic Activity --- p.63 / Chapter 2.2.14 --- Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) --- p.67 / Chapter 2.2.14.1 --- RNA Extraction by TRIzol Reagent --- p.67 / Chapter 2.2.14.2 --- Reverse Transcription --- p.68 / Chapter 2.2.14.3 --- Polymerase Chain Reaction --- p.68 / Chapter 2.3 --- Statistic Analysis --- p.71 / Chapter Chapter 3 --- Cytotoxicity of HK18 --- p.72 / Chapter 3.1 --- Cytotoxicity of HK18 on HepG2 Cells --- p.73 / Chapter 3.1.1 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by MTT Assay --- p.73 / Chapter 3.1.2 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by Tryphan Blue Exclusion Assay --- p.76 / Chapter 3.2 --- Cytotoxicity of HK18 on R-HepG2 Cells --- p.78 / Chapter 3.2.1 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by MTT Assay --- p.78 / Chapter 3.2.2 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by Tryphan Blue Exclusion Assay --- p.81 / Chapter 3.3 --- Cytotoxicity of HK18 on Macrophages --- p.83 / Chapter 3.4 --- Hemolytic Activity of HK18 --- p.85 / Chapter 3.5 --- In vivo Study of the Toxicity of HK18 --- p.87 / Chapter Chapter 4 --- Mechanistic Study of HK18 on HepG2 Cells --- p.90 / Chapter 4.1 --- Hallmarks of Apoptosis Induced by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.2 --- Induction of DNA Fragmentation by HK18 of HepG2 Cells --- p.97 / Chapter 4.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in HepG2 Cells --- p.99 / Chapter 4.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of HepG2 Cells --- p.99 / Chapter 4.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in HepG2 Cells --- p.101 / Chapter 4.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on HepG2 Cells --- p.105 / Chapter 4.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated HepG2 Cells --- p.108 / Chapter 4.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated HepG2 Cells --- p.114 / Chapter 4.2.1.5 --- HK18 Induced Cytochrome c and AIF Released from Mitochondria of HepG2 Cells --- p.120 / Chapter 4.3 --- Downstream Biochemical Changes Induced by HK18 on HepG2 Cells --- p.123 / Chapter 4.3.1 --- Activation of Caspase 3 of HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.123 / Chapter 4.3.2 --- Induction of Caspases Activities of HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.125 / Chapter 4.4 --- Down-regulation of Anti-apoptotic Bcl-2 Family Members of HepG2 Cells by HK18 --- p.129 / Chapter Chapter 5 --- Mechanistic Study of HK18 on R-HepG2 Cells --- p.133 / Chapter 5.1 --- Hallmarks of Apoptosis Induced by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.2 --- Induction of DNA Fragmentation by HK18 of R-HepG2 Cells --- p.137 / Chapter 5.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in R-HepG2 Cells --- p.139 / Chapter 5.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of R-HepG2 Cells --- p.139 / Chapter 5.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in R-HepG2 Cells --- p.139 / Chapter 5.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on R-HepG2 Cells --- p.142 / Chapter 5.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated R-HepG2 Cells --- p.144 / Chapter 5.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated R-HepG2 Cells --- p.146 / Chapter 5.3 --- Downstream Biochemical Changes Induced by HK18 on R-HepG2 Cells --- p.148 / Chapter 5.3.1 --- Activation of Caspase 3 of R-HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.148 / Chapter 5.3.2 --- Induction of Caspases Activation of R-HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.150 / Chapter 5.4 --- Down-regulation of the Anti-apoptotic Bcl-2 Protein of R-HepG2 Cells by HK18 --- p.154 / Chapter 5.5 --- HK18 was Not a Substrate of P-glycoprotein Contents --- p.156 / Chapter Chapter 6 --- Discussion --- p.158 / Chapter 6.1 --- Cytotoxicity of HK18 --- p.159 / Chapter 6.1.1 --- HK18 was Cytotoxic to the Human Hepatocellular Carcinoma Cell Line HepG2 and Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.159 / Chapter 6.1.2 --- Study of the Toxicity of HK18 --- p.160 / Chapter 6.2 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.1 --- Apoptotic Cell Death Induction of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.162 / Chapter 6.3 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.1 --- Apoptotic Cell Death Induction of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.181 / Chapter Chapter 7 --- Future Perspectives --- p.190 / Chapter Chapter 8 --- References --- p.193

by Ko Samuel. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 209-227). / Abstract also in Chinese. / Acknowledgements --- p.i / Abbreviations --- p.ii / Abstract --- p.vii / Abstract in Chinese --- p.x / List of Figures --- p.xiii / List of Tables --- p.xx / Publication --- p.xxi / Contents --- p.xxii / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter 1.1 --- Tumor Necrosis Factor --- p.2 / Chapter 1.1.1 --- History of Tumor Necrosis Factor --- p.2 / Chapter 1.1.2 --- TNF Subtypes and Their Purification --- p.3 / Chapter 1.1.3 --- Release of TNF --- p.9 / Chapter 1.1.4 --- Biological Actions of TNF --- p.9 / Chapter 1.2 --- Tumor Necrosis Factor Receptor --- p.11 / Chapter 1.2.1 --- Purification of TNF Receptor --- p.11 / Chapter 1.2.2 --- Regulation of TNF Receptor --- p.14 / Chapter 1.2.3 --- "Functions of TNF Receptor 1,Receptor 2 and Soluble TNF Receptors" --- p.15 / Chapter 1.3 --- Possible Signal Transductions of Tumor Necrosis Factor-Alpha --- p.17 / Chapter 1.3.1 --- Activation of Phospholipase A2 Cascade --- p.18 / Chapter 1.3.2 --- Activation of Phospho lipase C Pathway --- p.19 / Chapter 1.3.3 --- Activation of Sphingomyelin Pathway --- p.20 / Chapter 1.3.4 --- Activation of Protein Kinase --- p.22 / Chapter 1.3.5 --- Activation of the Cascade of Death Domain --- p.23 / Chapter 1.4 --- Induction of Both Necrosis and Apoptosis by Tumor Necrosis Factor-Alpha --- p.25 / Chapter 1.4.1 --- Apoptosis Versus Necrosis --- p.25 / Chapter 1.4.2 --- TNF Can Induce Both Apoptosis and Necrosis --- p.27 / Chapter 1.5 --- Possible Mechanisms of Tumor Necrosis Factor-Alpha- Mediated Cytotoxicity --- p.27 / Chapter 1.5.1 --- Release of Reactive Oxygen Species --- p.28 / Chapter 1.5.2 --- Release of Intracellular Calcium --- p.31 / Chapter 1.5.3 --- Miscellaneous Mechanisms --- p.36 / Chapter 1.6 --- Objective of Studies --- p.37 / Chapter Chapter 2. --- Materials and Methods --- p.39 / Chapter 2.1 --- Materials --- p.40 / Chapter 2.1.1 --- Buffer --- p.40 / Chapter 2.1.2 --- Culture Media --- p.45 / Chapter 2.1.3 --- Chemicals --- p.46 / Chapter 2.1.4 --- Culture of Cells --- p.49 / Chapter 2.1.4.1 --- "Tumor Necrosis Factor-Alpha-Sensitive Cell Line, L929" --- p.49 / Chapter 2.1.4.2 --- "Tumor Necrosis Factor-Alpha-Resistant Cell Line, rL929, rL929-l IE and rL929-4F" --- p.50 / Chapter 2.2 --- Methods --- p.50 / Chapter 2.2.1 --- Agarose Gel Electrophoresis --- p.50 / Chapter 2.2.2 --- Cytotoxicity Assay --- p.52 / Chapter 2.2.3 --- Confocal Laser Scanning Microscopy --- p.53 / Chapter 2.2.4 --- Flow Cytometry --- p.57 / Chapter Chapter 3. --- Results --- p.65 / Chapter 3.1 --- Induction of Apoptosis in Tumor Necrosis Factor-Alpha- Treated L929 Cell --- p.66 / Chapter 3.1.1 --- Introduction --- p.66 / Chapter 3.1.2 --- TNF Induced DNA Fragmentation in L929 Cells --- p.67 / Chapter 3.2 --- Effect of Tumor Necrosis Factor-Alpha on Cell Cycle --- p.73 / Chapter 3.2.1 --- Introduction --- p.73 / Chapter 3.2.2 --- Effect of TNF on Cell Cycle --- p.75 / Chapter 3.3 --- Release of Reactive Oxygen Species in Tumor Necrosis Factor-Alpha Treatment --- p.79 / Chapter 3.3.1 --- Introduction --- p.79 / Chapter 3.3.2 --- Release of Reactive Oxygen Species in TNF- Treated L929 Cells is Time Dependent --- p.81 / Chapter 3.3.3 --- Effect of Antioxidants on TNF-Mediated Cytotoxicity --- p.93 / Chapter 3.3.4 --- Effect of Mitochondrial Inhibitors on TNF-Mediated Cytotoxicity --- p.96 / Chapter 3.4 --- The Role of Calcium in Tumor Necrosis Factor-Alpha Treatment --- p.112 / Chapter 3.4.1 --- Introduction --- p.112 / Chapter 3.4.2 --- Release of Intracellular Calcium in TNF-Treated L929 Cells --- p.113 / Chapter 3.4.3 --- Effect of Calcium-Inducing Agents on TNF-Treated L929Cells --- p.127 / Chapter 3.5 --- Relationship between Reactive Oxygen Species and Calcium in Tumor Necrosis Factor-Alpha-Mediated Cytotoxicity --- p.133 / Chapter 3.5.1 --- Introduction --- p.133 / Chapter 3.5.2 --- Effect of Intracellular Calcium Chelator on TNF- Mediated ROS Release and Cytotoxicity --- p.133 / Chapter 3.5.3 --- Effect of Mitochondrial Calcium on TNF-Mediated ROS Release and Cytotoxicity --- p.147 / Chapter 3.6 --- Effect of Tumor Necrosis Factor-Alpha on pH --- p.162 / Chapter 3.6.1 --- Introduction --- p.162 / Chapter 3.6.2 --- Effect of TNF on pH --- p.162 / Chapter 3.7 --- Effect of Tumor Necrosis Factor-Alpha on Mitochondrial Membrane Potential --- p.165 / Chapter 3.7.1 --- Introduction --- p.165 / Chapter 3.7.2 --- Effect of TNF and Some Drugs on Mitochondrial Membrane Potential --- p.165 / Chapter 3.8 --- "Comparison of Effects of Tumor Necrosis Factor-Alpha on Susceptible Cell Line, L929 and Resistant Cell Line, rL929, rL929-11E and rL929-4F" --- p.169 / Chapter 3.8.1 --- Introduction --- p.169 / Chapter 3.8.2 --- Effect of TNF on the Cytotoxicity of Resistant Cell Lines --- p.170 / Chapter 3.8.3 --- Effect of TNF on the Release of ROS in Resistant Cell Lines --- p.170 / Chapter 3.8.4 --- Effect of TNF on the Release of Calcium in Resistant Cell Lines --- p.178 / Chapter 3.8.5 --- Effect of TNF on Cell Cycle in Resistant Cell Lines --- p.185 / Chapter Chapter 4. --- General Discussion --- p.187 / Chapter 4.1 --- Tumor Necrosis Factor Induced Apoptosis in L929 Cells --- p.188 / Chapter 4.2 --- Tumor Necrosis Factor Increased the Release of Reactive Oxygen Species in L929 Cells --- p.189 / Chapter 4.3 --- Tumor Necrosis Factor Increased the Release of Calcium in L929 Cells --- p.194 / Chapter 4.4 --- Calcium Induced Reactive Oxygen Species Release in TNF- Treated L929 Cells --- p.197 / Chapter 4.5 --- Tumor Necrosis Factor Did Not Change the pH and Mitochondrial Membrane Potential in TNF-Treated L929 Cells --- p.198 / Chapter 4.6 --- Tumor Necrosis Factor Did Not Increase the Release of Reactive Oxygen Species or Calcium in Resistant Cell Lines --- p.201 / Chapter Chapter 5. --- Future Perspective --- p.204 / Chapter 5.1 --- The Relationship Between Tumor Necrosis Factor and Cytochrome c --- p.205 / Chapter 5.2 --- The Relationship Between Tumor Necrosis Factor and Mitochondrial DNA Damage --- p.206 / Chapter 5.3 --- Clinical studies with Tumor Necrosis Factor --- p.206 / References --- p.208

by Chan Hung Chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 143-151). / Abstract also in Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / Abstract (Chinese version) --- p.v / Abbreviations --- p.viii / Content --- p.ix / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1. --- Free radical and free radical-mediated antitumor drugs --- p.1 / Chapter 2. --- Mitomycin C (MC) / Chapter 2.1 --- Drug actions of MC --- p.2 / Chapter 2.2 --- Adverse side effects of MC --- p.5 / Chapter 3. --- Doxorubicin (DOX) / Chapter 3.1 --- Drug actions of DOX --- p.7 / Chapter 3.2 --- Adverse side effects of DOX --- p.8 / Chapter 4. --- Antioxidants --- p.14 / Chapter 5. --- Effects of exogenous ATP on the antitumor activity of Doxorubicin and Mitomycin C / Chapter 5.1 --- Glutathione (GSH) and related enzymes --- p.17 / Chapter 5.2 --- Glutathione (GSH) and Anticancer Quinones --- p.19 / Chapter 5.3 --- Glutathione and the cardiac toxicity of the anticancer drugs --- p.20 / Chapter 5.4 --- Glutathione depletion in tumor cells by exogenous ATP --- p.21 / Chapter 6. --- Aim of research --- p.24 / Chapter CHAPTER TWO --- THE EFFECT OF ANTIOXIDANTS ON DOXORUBICIN- OR MITOMYCIN C-INDUCED CYTOTOXICITY ON HUMAN TUMOR AND NORMAL CELL LINES / Chapter 2.1 --- Introduction --- p.26 / Chapter 2.2 --- Materials and Methods --- p.28 / Chapter 2.3 --- Results --- p.36 / Chapter 2.4 --- Discussion --- p.60 / Chapter CHAPTER THREE --- STUDY OF CARDIOPROTECTIVE EFFECTS OF ANTIOXIDANTS AGAINST DOXORUBICIN- OR MITOMYCIN C-INDUCED TOXICITY BY LANGENDORFF PERFUEED ISOLATED RAT HEART MODEL / Chapter 3.1 --- Introduction --- p.64 / Chapter 3.2 --- Materials and Methods --- p.67 / Chapter 3.3 --- Results --- p.75 / Chapter 3.4 --- Discussion --- p.76 / Chapter CHAPTER FOUR --- THE EFFECT OF ANTIOXIDANTS DURING CHEMOTHERAPY OF DOXORUBICIN OR MITOMYCIN C IN TUMOR-BEARING MICE / Chapter 4.1 --- Introduction --- p.78 / Chapter 4.2 --- Materials and Methods --- p.80 / Chapter 4.3 --- Results --- p.83 / Chapter 4.4 --- Discussion --- p.93 / Chapter CHAPTER FIVE --- HISTOLOGICAL STUDY AND LIPID PEROXIDATION STUDY OF PROTECTIVE EFFECT OF ANTIOXIDANTS IN TUMOR-BEARING MICE TREATED WITH DOXORUBICIN OR MITOMYCIN C / Chapter 5.1 --- Introduction --- p.95 / Chapter 5.2 --- Materials and Methods --- p.98 / Chapter 5.3 --- Results --- p.103 / Chapter 5.4 --- Discussion --- p.117 / Chapter CHAPTER SIX --- EFFECT OF EXOGENOUS ATP ON THE ANTITUMOR ACTIVITY OF DOXORUBICIN AND MITOMYCIN C ON CULTURED HUMAN HEPATOMA CELLS / Chapter 6.1 --- Introduction --- p.122 / Chapter 6.2 --- Materials and Methods --- p.124 / Chapter 6.3 --- Results --- p.126 / Chapter 6.4 --- Discussion --- p.136 / Chapter CHAPTER SEVEN --- CONCLUSION / Chapter 7.1 --- Conclusion --- p.139 / Chapter 7.2 --- Future perspective --- p.141 / Bibliography --- p.142

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

Thesis (doctoral)--University of Lund, 1996. / Added t.p. with thesis statement added.

Thesis (doctoral)--University of Lund, 1996. / Added t.p. with thesis statement added.

Made available in DSpace on 2016-02-05T18:30:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-15. Added 1 bitstream(s) on 2016-02-05T18:34:15Z : No. of bitstreams: 1 000857711.pdf: 1685122 bytes, checksum: 06044cf174850099dc014984d0007cc8 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A prótese bucomaxilofacial implantorretida pode ser suportada por pele ou por mucosa. Subprodutos dos materiais utilizados na confecção destas próteses podem atuar como irritantes ou causadores de reações alérgicas a tais tecidos. A proposta do presente estudo foi avaliar a citotoxicidade de primers e adesivo utilizados na confecção de próteses maxilofaciais retidas por implantes, por meio da análise da proliferação celular e da produção de citocinas pró-inflamatórias e de proteínas de matriz extracelular por queratinócitos humanos. Foram confeccionadas 28 amostras de resina e silicone, em forma de discos (10 x 1 mm), unidas ou não pela aplicação de primer e/ou adesivo. Estas amostras foram distribuídas em 7 grupos: Resina (R), Silicone (S), Resina + Silastic Medhical Adhesive Type A + Silicone (RAS), Resina + DC 1205 primer + Silicone (RDCpS), Resina + Sofreliner primer + Silicone (RSpS), Resina + DC 1205 primer + Silastic Medhical Adhesive Type A + Silicone (RDCpAS) e Resina + Sofreliner primer + Silastic Medhical Adhesive Type A + Silicone (RSpAS). Os extratos dos materiais testados foram preparados colocando-se quatro amostras de cada grupo experimental em tubos de ensaio contendo 9 mL de meio de cultura DMEM (Dulbecco's Modified Eagle's) e incubados a 37°C por 24 horas. Após o período de incubação, a citotoxicidade dos extratos foi avaliada pelo ensaio de MTT em cultura de queratinócitos humanos (HaCaT). Foi avaliada, também, a produção das citocinas IL-1, IL-6 e TNF-α e a quimiocina MIP-1α por meio do ensaio ELISA (Ensaio Imunoabsorbente de Ligação Enzimática). Também foi avaliada a expressão de RNAm para MMP-9, TGF-β e COL-IV por meio da técnica de RT-PCR em tempo real (Reação em Cadeia da Polimerase em Tempo Real). Os dados obtidos foram submetidos à Análise de Variância (ANOVA), seguido... / The implant-retained maxillofacial prosthesis can be supported by skin and mucosa. The sub-products produced by the materials used to fabricate these prostheses may act as an irritant factor and cause allergy in these tissues. The aim of this study was to evaluate the cytotoxic effect of primers and adhesive used to bond the acrylic resin and the facial silicone during implant-retained maxillofacial prosthesis fabrication, through the analysis of the cell proliferation, and the production of proinflammatory cytokines and extracellular matrix proteins by keratinocytes. A total of 28 round shape samples (10 x 1 mm) made of resin and silicone bonded or not with primer and adhesive was fabricated. Samples were divided into 7 groups: Resin (R), Silicone (S), Resin + Silastic Medical Adhesive Type A + Silicone (RAS), Resin + DC 1205 primer + Silicone (RDCpS), Resin + Sofreliner primer + Silicone (RSpS), Resin + DC 1205 primer + Silastic Medical Adhesive Type A + Silicone (RDCpAS), and Resin + Sofreliner primer + Silastic Medical Adhesive Type A + Silicone (RSpAS). Extracts of tested materials were prepared setting four samples of each experimental group in Falcon tube with 9mL of medium (Bulbecco's Modified Eagle's) and incubated at 37°C for 24 hours. After incubation period, the extract cytotoxicity was evaluated by an assay of cell survival/proliferation (MTT test) in cultures of human keratinocytes (HaCaT). The levels of IL-1, IL-6 and TNF-α and the chemokine MIP-1α were evaluated by ELISA (Enzyme-Linked Immunoabsorbent Assay). The mRNA expression for MMP-9, TGF-β and collagen type IV were analyzed by the RT-PCR (Real time polymerase chain reaction). Data were submitted to the analysis of variance with Bonferroni post-tests (p<0.05). The results showed increased cell proliferation for the RAS group. The RDCpS group showed the highest...

Orientadora: Daniela Micheline dos Santos / Coorientador: Marcelo Coelho Goiato / Banca: Marcela Fillié Haddad / Banca: Sandra Helena Penha de Oliveira / Resumo: A prótese bucomaxilofacial implantorretida pode ser suportada por pele ou por mucosa. Subprodutos dos materiais utilizados na confecção destas próteses podem atuar como irritantes ou causadores de reações alérgicas a tais tecidos. A proposta do presente estudo foi avaliar a citotoxicidade de primers e adesivo utilizados na confecção de próteses maxilofaciais retidas por implantes, por meio da análise da proliferação celular e da produção de citocinas pró-inflamatórias e de proteínas de matriz extracelular por queratinócitos humanos. Foram confeccionadas 28 amostras de resina e silicone, em forma de discos (10 x 1 mm), unidas ou não pela aplicação de primer e/ou adesivo. Estas amostras foram distribuídas em 7 grupos: Resina (R), Silicone (S), Resina + Silastic Medhical Adhesive Type A + Silicone (RAS), Resina + DC 1205 primer + Silicone (RDCpS), Resina + Sofreliner primer + Silicone (RSpS), Resina + DC 1205 primer + Silastic Medhical Adhesive Type A + Silicone (RDCpAS) e Resina + Sofreliner primer + Silastic Medhical Adhesive Type A + Silicone (RSpAS). Os extratos dos materiais testados foram preparados colocando-se quatro amostras de cada grupo experimental em tubos de ensaio contendo 9 mL de meio de cultura DMEM (Dulbecco's Modified Eagle's) e incubados a 37°C por 24 horas. Após o período de incubação, a citotoxicidade dos extratos foi avaliada pelo ensaio de MTT em cultura de queratinócitos humanos (HaCaT). Foi avaliada, também, a produção das citocinas IL-1, IL-6 e TNF-α e a quimiocina MIP-1α por meio do ensaio ELISA (Ensaio Imunoabsorbente de Ligação Enzimática). Também foi avaliada a expressão de RNAm para MMP-9, TGF-β e COL-IV por meio da técnica de RT-PCR em tempo real (Reação em Cadeia da Polimerase em Tempo Real). Os dados obtidos foram submetidos à Análise de Variância (ANOVA), seguido... / Abstract: The implant-retained maxillofacial prosthesis can be supported by skin and mucosa. The sub-products produced by the materials used to fabricate these prostheses may act as an irritant factor and cause allergy in these tissues. The aim of this study was to evaluate the cytotoxic effect of primers and adhesive used to bond the acrylic resin and the facial silicone during implant-retained maxillofacial prosthesis fabrication, through the analysis of the cell proliferation, and the production of proinflammatory cytokines and extracellular matrix proteins by keratinocytes. A total of 28 round shape samples (10 x 1 mm) made of resin and silicone bonded or not with primer and adhesive was fabricated. Samples were divided into 7 groups: Resin (R), Silicone (S), Resin + Silastic Medical Adhesive Type A + Silicone (RAS), Resin + DC 1205 primer + Silicone (RDCpS), Resin + Sofreliner primer + Silicone (RSpS), Resin + DC 1205 primer + Silastic Medical Adhesive Type A + Silicone (RDCpAS), and Resin + Sofreliner primer + Silastic Medical Adhesive Type A + Silicone (RSpAS). Extracts of tested materials were prepared setting four samples of each experimental group in Falcon tube with 9mL of medium (Bulbecco's Modified Eagle's) and incubated at 37°C for 24 hours. After incubation period, the extract cytotoxicity was evaluated by an assay of cell survival/proliferation (MTT test) in cultures of human keratinocytes (HaCaT). The levels of IL-1, IL-6 and TNF-α and the chemokine MIP-1α were evaluated by ELISA (Enzyme-Linked Immunoabsorbent Assay). The mRNA expression for MMP-9, TGF-β and collagen type IV were analyzed by the RT-PCR (Real time polymerase chain reaction). Data were submitted to the analysis of variance with Bonferroni post-tests (p<0.05). The results showed increased cell proliferation for the RAS group. The RDCpS group showed the highest... / Mestre