Tumour necrosis has long been associated with poor prognosis and reduced survival in cancer. Hypotheses to explain this include the idea that as aggressive tumours tend to grow rapidly, they outgrow their blood supply leading to areas of hypoxia and subsequently necrosis. However whilst this and similar hypotheses have been put forward to explain the association, the biological significance of the cells which make up necrotic tissue has been largely ignored. This stems from the belief that because a tumour is more aggressive and fast growing it develops areas of necrosis, rather than, the tumour is more aggressive because it contains areas of necrosis. Which came first like the egg and chicken is yet to be determined, however to date most research has only considered the possibility of the former. Viable cells were found in the necrotic core of Multicellular Tumour Spheroids. When examined these cells were found to be different to the original cell line in terms of proliferation, migration, and chemosensitivity. A proteomic analysis showed that these phenotypical changes were accompanied by changes in a large number of proteins within the cells, some of which could be potential therapeutic targets. Furthermore this has led to a new hypothesis for tumour necrosis and its association with poor prognosis. Necrotic tissue provides a microenvironemental niche for cells with increased survival capabilities. Protected from many chemotherapeutics by their non-proliferative status once conditions improve these cells can return to proliferation and repopulate the tumour with an increasingly aggressive population of cells. / Yorkshire Cancer Research

No description available.

No description available.

No description available.

Several epidemiological studies have suggested a role for environmental chemicals in the etiology of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Endosulfan (an organochlorine) and zineb (zinc-ethylene-bis-dithiocarbamate) are used as pesticides on a variety of crops worldwide and pose potential health risks to humans and animals. Both endosulfan and zineb are known to affect nervous system. Because the dopaminergic system continues to develop postnatally, we hypothesized that developmental exposure to endosulfan or zineb alone or in combination would result in alteration of nigrostrial neurotransmitters and would render the nigrostrial dopamine system more susceptible to chemical challenge later in life. The objectives of this study were (1) to determine the effects of endosulfan and zineb individually and in combination on dopaminergic or cholinergic pathways in vivo, (2) to investigate the effects of exposure to endosulfan, zineb and their mixtures administered in early life (during brain development) on subsequent exposure to these pesticides on the dopaminergic and cholinergic systems, in vivo, (3) to investigate the mechanism(s) of induction of neuronal cell death caused by these pesticides using human neuroblastoma SH-SY5Y cells in culture, (4) to define the role of oxidative stress in pesticide-induced neuronal cell death in vitro. Male C57Bl/6 mice of 7-9 months old exposed to zineb (50 and 100 mg/kg), endosulfan (1.55, 3.1 and 6.2 mg/kg) and their mixtures every other day over a 2-week period exhibited higher levels of dopamine accumulation in the striatum. Both pesticide-treated groups displayed significantly lower norepinephrine levels in the striatum (Ï ≤ 0.05) than the controls. The developmental exposure to zineb, endosulfan and their combination enhanced the vulnerability to subsequent neurotoxic challenges occurring later in life. Thus, C57BL/6 mice exposed to zineb, endosulfan and their mixtures as juveniles (postnatal days 5 to 19) and re-exposed at 8 months of age showed a significant depletion of striatal dopamine, to 22%, 16%, and 35% of control, respectively. Acetylcholinesterase activity in the cerebral cortex was found to be significantly increased in all pesticide treated groups. Mice given mixtures of pesticides also showed significantly increased levels of normal and aggregated alpha-synuclein, a hallmark of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. The results of these studies indicate that exposure to these pesticides as neonates and re-exposure as adults could result in neurochemical changes that did not reveal at adulthood when the exposure was at juvenile age only. We further investigated the mechanism(s) of activation of pesticide-induced neuronal cell death in vitro. The characteristic of cell death in SH-SY5Y human neuroblastoma cells was examined. These cells are known to retain catecholaminergic phenotype. Cells were exposed to endosulfan, zineb and mixtures of two pesticides, in concentrations ranging from 50 μM to 400 μM. These exposures caused both apoptotic and necrotic cell death in SH-SY5Y cells as evaluated by lactate dehydrogenase release, 7-aminoactinomycin-D and Annexin-V/PI assays. Exposure to mixtures of the pesticides enhanced both the early apoptosis and late apoptosis/necrosis compared to either chemical alone. Visual evaluation using DNA ladder assay and fluorescence Annexin V/PI assay confirmed the contribution of both apoptotic and necrotic events. Furthermore, endosulfan and zineb alone and in combination altered the caspase-3 activity indicating that both pesticides exposure exert their apoptotic effect via the caspase-3 pathway. Because there has been increasing evidence of the role of reactive oxygen species (ROS) and oxidative stress in pesticide-induced neuronal cell death (apoptosis and necrosis), the levels of ROS and antioxidant enzymes were examined. Cells treated with pesticides were found to enhance the generation of superoxide anion and hydrogen peroxide both in a dose- and time-dependent manner. Mixture of pesticides significantly enhanced the production of these reactive oxygen species compared to cells exposed to individual pesticide. Cells treated with pesticides showed a decrease in superoxide dismutase, glutathione peroxidase, and catalase levels. These pesticides also induced lipid peroxides (thiobarbituric acid reactive products) formation in SH-SY5Y cells. Furthermore, cells exposed to these pesticides were found to have increased in the expression of NFkappaB activity in the nucleus. These data support the hypothesis that oxidative stress was induced in neuronal cells by exposing to these pesticides in vitro. Taken together, the results of this study support the above hypothesis and suggest that the cytotoxicity of endosulfan and zineb and their combinations may, at least in part, be associated with the generation of ROS. Furthermore, mice exposed at early age and re-exposed at adulthood become more susceptible to alteration of neurotransmitter levels compared to mice exposed to these pesticides only as juveniles. These findings could add to the growing body of knowledge on the mechanism of pesticide-induced dopaminergic neuronal cell death and could hold tremendous implication for the future understanding of the possible involvement of environmental risk factors in the pathogenesis of Parkinson's disease. / Ph. D.

Tsoi Lo Ming. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 99-109). / Abstracts in English and Chinese. / Acknowledgements --- p.I / Abstract --- p.II / Abstract in Chinese --- p.III / List of Abbreviations --- p.IV / List of Figures --- p.V / List of Tables --- p.VI / Table of Contents --- p.VII / Chapter Chapter 1 : --- Introduction / Chapter 1.1. --- Apolipoprotein and Lipoprotein Metabolism --- p.1 / Chapter 1.2. --- Molecular Information of ApoE --- p.2 / Chapter 1.3. --- Tissue Distribution of ApoE --- p.2 / Chapter 1.4. --- Functions of ApoE --- p.4 / Chapter 1.5. --- Genetic Polymorphism of ApoE --- p.7 / Chapter 1.6. --- Protein Structure and Characteristics of ApoE Isoforms --- p.9 / Chapter 1.7. --- Plasma and Cellular Expression Level of ApoE Isoforms --- p.12 / Chapter 1.8. --- Association between ApoE Isoforms and Plasma Lipid Profiles --- p.13 / Chapter 1.9. --- ApoE Polymorphisms and Pathophysiological Conditions / Chapter 1.9.1. --- Type III Hyperlipoproteinemia (Type III HLP) --- p.14 / Chapter 1.9.2. --- Alzheimer's Disease --- p.15 / Chapter 1.9.3. --- Atherosclerosis / Chapter 1.9.3.1. --- Atherosclerosis - An Inflammatory Process --- p.15 / Chapter 1.9.3.2. --- Role of ApoE in Atherosclerosis --- p.18 / Chapter (a) --- Functions Associated to Lipid Metabolism --- p.19 / Chapter (b) --- Functions Independent to Lipid Metabolism --- p.20 / Chapter 1.9.3.3. --- TNF-α and IL-6 in Atherosclerosis --- p.25 / Chapter 1.10. --- Macrophage Cytokine Expression and MAPKs / Chapter 1.10.1. --- Organization of MAPKs Signaling Pathway --- p.26 / Chapter 1.10.2. --- Lipopolysaccharide and MAPKs in Macrophage Cytokine Expression --- p.28 / Chapter 1.10.3. --- Regulation of Macrophage Cytokine Expression / Chapter 1.10.3.1. --- ERK1/2 and p38 MAPK Pathway --- p.30 / Chapter 1.10.3.2. --- Arachidonic Acid Metabolism --- p.30 / Chapter 1.11. --- Aim and Hypothesis --- p.31 / Chapter Chapter 2 : --- Materials and Methods / Materials / Chapter 2.1 --- Culture of ApoE-isoform-expressing J774A.1 Macrophage Cell Line --- p.32 / Chapter 2.2 --- RNA Extraction and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.33 / Chapter 2.3 --- Protein Extraction and Quantification --- p.37 / Chapter 2.4 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.38 / Chapter 2.5 --- Western Blotting --- p.39 / Chapter 2.6 --- LPS Treatment --- p.42 / Chapter 2.7 --- MAPK Inhibitor Experiment --- p.43 / Methods / Chapter 2.8 --- Study on the Effect of Endogenously Expressed ApoE Isoforms on Macrophage Cytokine Secretion / Chapter 2.8.1. --- Establishment of ApoE-isoform-expressing Macrophages --- p.44 / Chapter 2.8.2. --- Semi-quantification of ApoE mRNA Level by RT-PCR / Chapter 1) --- Isolation of Total RNA --- p.45 / Chapter 2) --- RT-PCR --- p.46 / Chapter 2.8.3. --- Determination of ApoE Protein Expression Level by ELISA and Western Blot --- p.47 / Chapter 1) --- Quantification of Total Proteins --- p.48 / Chapter 2) --- ELISA --- p.48 / Chapter 3) --- Western Blot --- p.49 / Chapter 2.8.4. --- LPS Treatment --- p.51 / Chapter 2.8.5. --- MEK1/2 Inhibitor Experiment --- p.53 / Chapter 2.8.6. --- p38 Inhibitor Experiment --- p.54 / Chapter 2.9 --- Study on the Effect of Exogenous ApoE Isoform on Macrophage Cytokine Secretion --- p.55 / Chapter 2.10 --- Statistical Analysis --- p.55 / Chapter Chapter 3: --- Results / Changes of Inflammatory Properties Associated with Endogenous ApoE Isoform Expression in Macrophages / Chapter 3.1 --- Characterization of ApoE-isoform-expressing Macrophages --- p.56 / Chapter 3.1.1. --- Cell Lines with Stable Expression of ApoE Isoforms --- p.56 / Chapter 3.2 --- Cell Morphology Study --- p.58 / Chapter 3.3 --- Changes of IL-6 and TNF-α Secretion Associated with Endogenous ApoE Isoforms Expression / Chapter 3.3.1. --- In the Presence of Lipoproteins --- p.60 / Chapter 3.3.2. --- Serum/Lipoprotein-independent Effects of ApoE Isoforms --- p.63 / Chapter 3.4 --- The Effects of Endogenous ApoE Isoform Expression on the Activities of MAPK Signaling Pathways / Chapter 3.4.1. --- Study on the Activation Status and Expression of MAPKs --- p.66 / Chapter 1) --- ERK1/2 MAPK Pathway --- p.66 / Chapter 2) --- p38 MAPK Pathway --- p.69 / Chapter 3.4.2. --- IL-6 and TNF-a Secretion Among ApoE Isoforms in the Presence of MEK1/2 mhibitor --- p.72 / Chapter 3.4.3. --- IL-6 and TNF-α Secretion Among ApoE Isoforms in the Presence of p38 Inhibitor --- p.75 / Chapter Chapter 4 : --- Discussions / Chapter 4.1. --- Mouse Peritoneal Macrophage Cell Line J774A.1 as Cell Model --- p.79 / Chapter 4.2. --- Inflammatory Properties Associated with Endogenous ApoE Isoform Expression in Macrophages / Chapter 4.2.1. --- Expression Level of ApoE Isoform Transgenes in Mouse Peritoneal Macrophages --- p.80 / Chapter 4.2.2. --- Macrophage Activation by LPS --- p.81 / Chapter 4.2.3. --- Effect of Endogenous ApoE Isoform Expression on Cytokine Secretion and Signal Transduction in Macrophages --- p.82 / Chapter 4.3. --- Conclusions and Future Prospects / Chapter 4.3.1. --- Conclusions --- p.90 / Chapter 4.3.2. --- Future Prospects --- p.91 / Chapter Chapter 5 : --- Appendices / Chapter 5.1 --- Changes of Inflammatory Properties of Macrophages Supplemented with Exogenous ApoE Isoforms / Chapter 5.1.1. --- Changes of IL-6 and TNF-a Secretion in Macrophages Supplemented with Exogenous ApoE Isoforms --- p.92 / Chapter 5.1.2. --- Changes of Signal Transduction in Macrophages Supplemented with Exogenous ApoE Isoforms / Chapter 5.1.2.1. --- Study on the Activation Status and Expression of MAPKs / Chapter 1) --- ERK1/2 MAPK Pathway --- p.95 / Chapter 2) --- p38 MAPK Pathway --- p.97 / Chapter Chapter 6: --- Bibliography --- p.99

by Yuen Wai Fan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 211-232). / Abstracts in English and Chinese. / Acknowledgements --- p.i / List of Publications and Abstracts --- p.ii / Abbreviations --- p.iv / Abstract --- p.xi / Abstract in Chinese --- p.xiv / List of Figures --- p.xvii / List of Tables --- p.xxiii / Contents --- p.xxiv / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter 1.1 --- Hyperthermia --- p.2 / Chapter 1.1.1 --- History of Hyperthermia --- p.2 / Chapter 1.1.2 --- Biological Functions of Hyperthermia --- p.3 / Chapter 1.1.3 --- Clinical Application of Hyperthermia --- p.4 / Chapter 1.1.3.1 --- Whole-body Hyperthermia --- p.4 / Chapter 1.1.3.2 --- Regional Hyperthermia --- p.4 / Chapter 1.1.3.3 --- Local Hyperthermia --- p.5 / Chapter 1.1.4 --- Combination Therapy --- p.5 / Chapter 1.1.4.1 --- Combined treatment with Hyperthermia and Radiotherapy --- p.6 / Chapter 1.1.4.2 --- Combined treatment with Hyperthermia and Chemotherapy --- p.6 / Chapter 1.2 --- Tumour Necrosis Factor --- p.9 / Chapter 1.2.1 --- History of Tumour Necrosis Factor --- p.9 / Chapter 1.2.2 --- Sources of TNF-α and TNF-β --- p.9 / Chapter 1.2.3 --- Biological Roles of TNF --- p.10 / Chapter 1.2.3.1 --- Receptors of TNF-α --- p.11 / Chapter 1.2.4 --- Signaling Pathway of TNF --- p.12 / Chapter 1.2.4.1 --- Activation of Death Domain --- p.12 / Chapter 1.2.4.2 --- Activation of Sphingomyelin Pathway --- p.13 / Chapter 1.2.4.3 --- Activation of NF-kB pathway --- p.13 / Chapter 1.3 --- Types of Cell Death: Necrosis and Apoptosis --- p.16 / Chapter 1.3.1 --- Necrosis --- p.16 / Chapter 1.3.2 --- Apoptosis --- p.16 / Chapter 1.4 --- Signaling Pathway in Apoptosis --- p.19 / Chapter 1.4.1 --- Factors Involved in Apoptotic Pathway --- p.19 / Chapter 1.4.1.1 --- Caspases --- p.19 / Chapter 1.4.1.2 --- Death Substrates --- p.20 / Chapter 1.4.1.3 --- Bcl-2 Protein Family --- p.21 / Chapter 1.4.1.4 --- Role of Mitochondria --- p.23 / Chapter 1.5 --- Objectives of the Project --- p.26 / Chapter Chapter 2. --- Materials and Methods --- p.28 / Chapter 2.1 --- Materials --- p.29 / Chapter 2.1.1 --- Culture of Cells --- p.34 / Chapter 2.1.1.1 --- "TNF-α Sensitive Cell Line, L929" --- p.34 / Chapter 2.1.1.2 --- "TNF-α Resistance Cell Line, L929-11E" --- p.34 / Chapter 2.1.1.3 --- Preservation of Cells --- p.35 / Chapter 2.1.2 --- Culture Media --- p.36 / Chapter 2.1.2.1 --- RPMI 1640 (Phenol Red Medium) --- p.36 / Chapter 2.1.2.2 --- RPMI 1640 (Phenol Red-Free Medium) --- p.36 / Chapter 2.1.3 --- Buffers and Reagents --- p.37 / Chapter 2.1.3.1 --- Preparation of Buffers --- p.37 / Chapter 2.1.3.2 --- Buffer for Common Use --- p.37 / Chapter 2.1.3.3 --- Reagents for Annexin-V-FITC/PI assay --- p.37 / Chapter 2.1.3.4 --- Reagents for Cytotoxicity Assay --- p.37 / Chapter 2.1.3.5 --- Reagents for Molecular Biology Work --- p.38 / Chapter 2.1.3.6 --- Reagents for Western Blotting Analysis --- p.38 / Chapter 2.1.4 --- Chemicals --- p.40 / Chapter 2.1.4.1 --- Recombinant Murine TNF-α --- p.40 / Chapter 2.1.4.2 --- Dye for Cytotoxicity Assay --- p.41 / Chapter 2.1.4.3 --- Fluorescence Dyes --- p.41 / Chapter 2.1.4.4 --- Chemicals Related to Mitochondrial Studies --- p.41 / Chapter 2.1.4.5 --- Inhibitors of Caspases --- p.42 / Chapter 2.1.4.6 --- Antibodies for Western Blotting --- p.42 / Chapter 2.1.4.7 --- Other Chemicals --- p.43 / Chapter 2.2 --- Methods --- p.44 / Chapter 2.2.1 --- Treatment with TNF-α --- p.44 / Chapter 2.2.2 --- Treatment with Hyperthermia --- p.44 / Chapter 2.2.3 --- In vitro Cell Cytotoxicity Assay --- p.45 / Chapter 2.2.4 --- Flow Cytometry --- p.46 / Chapter 2.2.4.1 --- Introduction --- p.46 / Chapter 2.2.4.2 --- Analysis by FCM --- p.48 / Chapter 2.2.4.3 --- Determination of Apoptotic and Late Apoptotic/Necrotic Cells with Annexin-V-FITC/PI Cytometric Analysis --- p.50 / Chapter 2.2.4.4 --- Determination of Mitochondrial Membrane Potential (ΔΨm) --- p.51 / Chapter 2.2.4.5 --- Determination of Hydrogen Peroxide (H202) Release --- p.52 / Chapter 2.2.4.6 --- Determination of Intracellular Free Calcium ([Ca2+]i) Level --- p.52 / Chapter 2.2.4.7 --- Determination of the Relationship of ΔΨm and [Ca2+]i Level --- p.53 / Chapter 2.2.5 --- Western Blotting Analysis --- p.53 / Chapter 2.2.5.1 --- Preparation of Proteins from Cells --- p.53 / Chapter 2.2.5.2 --- SDS Polyacrylamide Gel Electophoresis (SDS- PAGE) --- p.56 / Chapter 2.2.5.3 --- Electroblotting of Proteins --- p.57 / Chapter 2.2.5.4 --- Probing Antibodies for Proteins --- p.57 / Chapter 2.2.5.5 --- Enhanced Chemiluminescence (ECL) assay --- p.58 / Chapter 2.2.6 --- Reverse Transcriptase Polymerase Chain Reaction --- p.58 / Chapter 2.2.6.1 --- Extraction of RNA by Trizol Reagent --- p.59 / Chapter 2.2.6.2 --- Determination of the Amount of RNA --- p.60 / Chapter 2.2.6.3 --- Agarose Gel Electrophoresis --- p.60 / Chapter 2.2.6.4 --- Reverse Transcription --- p.63 / Chapter 2.2.6.5 --- Polymerase Chain Reaction (PCR) --- p.63 / Chapter 2.2.6.6 --- Design of Primers for Different Genes --- p.64 / Chapter 2.2.6.7 --- Determination of the Number of Cycles in PCR for Different Genes --- p.67 / Chapter 2.2.7 --- Caspase Fluorescent Assay --- p.67 / Chapter 2.2.7.1 --- Caspase-3 or ´ؤ8 Assay --- p.67 / Chapter Chapter 3. --- Results --- p.59 / Chapter 3.1 --- Studies of the Characteristics of L929 and L929-11E cells --- p.70 / Chapter 3.1.1 --- Determination of the Growth Curve of L929 and L929-11E Cells --- p.70 / Chapter 3.2 --- Studies on the Effect of TNF-α on L929 and L929-11E Cells --- p.73 / Chapter 3.2.1 --- TNF-α Induced Cell Death in L929 Cells but not in L929- 11E Cells --- p.73 / Chapter 3.2.2 --- TNF-α Induced Apoptosis in a Time-dependent Manner in L929Cells but not in L929-11E Cells --- p.80 / Chapter 3.2.3 --- TNF-α Induced Mitochondrial Membrane Depolarization in a Time-dependent Manner in L929 Cells but notin L929-11E Cells --- p.87 / Chapter 3.2.4 --- TNF-α Induced Cytochrome c Release in a Time- dependent Manner in L929 Cells but not in L929-11E Cells --- p.92 / Chapter 3.3 --- Effect of Hyperthermia on L929 and L929-11E Cells --- p.96 / Chapter 3.3.1 --- Introduction --- p.95 / Chapter 3.3.2 --- Hyperthermia Induced Apoptosis in L929 and L929-11E Cells --- p.96 / Chapter 3.3.3 --- Effect of Hyperthermia on Mitochondrial Membrane Depolarization --- p.100 / Chapter 3.3.4 --- Hyperthermia Induced Cyto c Release in a Time-dependent Manner in L929 and L929-11E Cells --- p.105 / Chapter 3.4 --- Relationship of Hyperthermia and TNF-α with PTP in L929 Cells --- p.107 / Chapter 3.5 --- Effect of TNF-α and Hyperthermia on the Level of Hydrogen Peroxide (H202) in L929 and L929-11E Cells --- p.114 / Chapter 3.5.1 --- Introduction --- p.114 / Chapter 3.5.2 --- TNF-α Enhanced the Level of H202 in L929 cells but not in L929-11E Cells --- p.115 / Chapter 3.5.3 --- Hyperthermia Enhanced the Level of H202 in L929 and L929-11E cells --- p.117 / Chapter 3.6 --- Effect of TNF-α and Hyperthermia on the Level of Intracellular Calcium in L929 and L929-11E Cells --- p.122 / Chapter 3.6.1 --- Increase in the Intracellular Calcium Level Induced by TNF-α Was Related to the Mitochondrial Membrane Depolarization in L929 Cells but not in L929-11E Cells --- p.122 / Chapter 3.6.2 --- Hyperthermia Increased the Level of [Ca2+]i in L929 and L929-11E Cells in a Time-dependent Manner --- p.124 / Chapter 3.7 --- Effect of Combined Hyperthermia and TNF-α Treatment on the Induction of Apoptosis in L929 and L929-1 1E Cells --- p.129 / Chapter 3.7.1 --- Combined Treatment with Hyperthermia and TNF- α Induced Apoptosis in Both L929 and L929-11E cells --- p.129 / Chapter 3.7.2 --- Hyperthermia and Its Combined Treatment with TNF-α Induced Mitochondrial Membrane Depolarization in L929 and L929-11E Cells --- p.135 / Chapter 3.8 --- Investigation of the Downstream Apoptotic Pathway in L929 and L929-11E Cells Upon Hyperthermia and TNF-a treatment --- p.142 / Chapter 3.8.1 --- Introduction --- p.142 / Chapter 3.8.2 --- Effect ofTNF-α and Hyperthermia on p53 Expression --- p.142 / Chapter 3.8.3 --- Effect of Hyperthermia and TNF-α on PARP --- p.146 / Chapter 3.8.4 --- Effect of Hyperthermia and TNF-α on Caspase-3 Activity --- p.149 / Chapter 3.8.5 --- Effect of Hyperthermia and TNF-α on Bid protein --- p.158 / Chapter 3.8.6 --- Effect of Hyperthermia and TNF-α on Caspase-8 Activity --- p.165 / Chapter 3.8.7 --- Effect ofTNF-α on TNFR1 Expression --- p.169 / Chapter Chapter 4. --- Discussion / Chapter 4.1 --- TNF-α Induced Apoptosis and Changed the Mitochondrial Activities in L929 Cells --- p.176 / Chapter 4.2 --- L929-11E cells Possessed Resistance Towards TNF-α --- p.187 / Chapter 4.3 --- Hyperthermia Triggered Apoptosis and Changed Mitochondrial Activities in L929 and L929-11E cells --- p.190 / Chapter 4.4 --- Combined hyperthermia and TNF-α treatment induced cell death and changed mitochondria activities in L929 and L929-11E cells --- p.195 / Chapter 4.5 --- Reversal of the TNF-α resistance and Enhancement of Sensitivity Towards Hyperthermia in L929-11E cells --- p.197 / Chapter 4.6 --- Proposed Pathway in the TNF-α- and Hyperthermia-mediated Apoptosis --- p.200 / Chapter 4.7 --- Application of TNF-α and Hyperthermia on Clinical Cancer Treatment --- p.203 / Chapter Chapter 5. --- Future Perspective of the Project --- p.206 / References --- p.210

Wong Chun Lung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 202-225). / Abstracts in English and Chinese. / Thesis Committee --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Acknowledgements --- p.ix / Table of Contents --- p.x / List of Abbreviations --- p.xv / List of Figures --- p.xix / List of Tables --- p.xx iii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- "Neurodegeneration, Inflammation and Gliosis" --- p.1 / Chapter 1.2 --- Anatomy of the CNS --- p.5 / Chapter 1.3 --- Astrocytes --- p.6 / Chapter 1.3.1 --- Morphology and Identification of Astrocytes --- p.6 / Chapter 1.3.2 --- Physiological Functions of Astrocytes in the CNS --- p.7 / Chapter 1.3.2.1 --- Induction of Blood-brain Barrier (BBB) --- p.7 / Chapter 1.3.2.2 --- Metabolism of Neurotransmitters --- p.9 / Chapter 1.3.2.3 --- Nursing Role of Astrocytes --- p.9 / Chapter 1.3.2.4 --- Immunological Functions of Astrocytes --- p.10 / Chapter 1.3.3 --- Neonatal Rat Cortical Astrocytes as In Vitro Model --- p.12 / Chapter 1.4 --- Cytokines in Brain Damage --- p.14 / Chapter 1.4.1 --- Lipopolysaccharides (LPS) --- p.16 / Chapter 1.4.2 --- Tumor Necrosis Factor-α (TNF-α) --- p.17 / Chapter 1.4.3 --- Interleukin-1 (IL-1) --- p.19 / Chapter 1.4.4 --- Interleukin-6 (IL-6) --- p.20 / Chapter 1.4.5 --- Interferon-γ (IFN-γ) --- p.21 / Chapter 1.5 --- Cytokines-induced Signaling Cascade --- p.22 / Chapter 1.5.1 --- TNF Receptors --- p.23 / Chapter 1.5.2 --- Ca2+ --- p.25 / Chapter 1.5.3 --- MAPK --- p.26 / Chapter 1.5.4 --- PICA --- p.27 / Chapter 1.5.5 --- NFkB --- p.29 / Chapter 1.6 --- Glucose Metabolism in the Brain and Glucose Transporters --- p.31 / Chapter 1.6.1 --- Glucose Transporters in the Brain --- p.32 / Chapter 1.6.2 --- Glucose Transporters in Brain Damage --- p.34 / Chapter 1.7 --- Ascorbic Acid Metabolism in the Brain --- p.36 / Chapter 1.8 --- Aim and Scope of this Project --- p.39 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Neonatal Sprawley 一Dawley Rats --- p.43 / Chapter 2.1.2 --- Plain Dulbecco Modified Eagle Medium ´ؤ Formula 12 (pDF12) --- p.43 / Chapter 2.1.3 --- Complete DF-12(cDF12) --- p.43 / Chapter 2.1.4 --- Phosphate Buffered Saline (PBS) --- p.44 / Chapter 2.1.5 --- Hank's Buffer (HSB) --- p.44 / Chapter 2.1.6 --- D/L-Homocysteine Buffer --- p.44 / Chapter 2.1.7 --- "LPS, Cytokines and Pentoxifylline" --- p.45 / Chapter 2.1.8 --- Specific TNF Receptor Agonist: TNF antibodies --- p.45 / Chapter 2.1.9 --- Calcium Modulators --- p.45 / Chapter 2.1.10 --- PKA Modulators --- p.46 / Chapter 2.1.11 --- NFkB Inhibitors --- p.47 / Chapter 2.1.12 --- MAPK Inhibitors --- p.47 / Chapter 2.1.13 --- β-Adrenergic Receptor Modulators --- p.47 / Chapter 2.1.14 --- Reagents for RNA and Protein Isolation --- p.48 / Chapter 2.1.15 --- Reagents for Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.48 / Chapter 2.1.16 --- Reagents for DNA Electrophoresis --- p.49 / Chapter 2.1.17 --- Reagents for Real-time PCR --- p.51 / Chapter 2.1.18 --- Reagents for Western Blotting --- p.51 / Chapter 2.1.19 --- Reagents for MTT Assay --- p.51 / Chapter 2.1.20 --- Reagents for 3H-Thymidine Incorporation Assay --- p.52 / Chapter 2.1.21 --- Reagents for Glucose Uptake Assay --- p.52 / Chapter 2.1.22 --- Reagents for Ascorbic Acid Accumulation Assay --- p.53 / Chapter 2.1.23 --- Reagents for Immunostammg --- p.53 / Chapter 2.1.24 --- Other Chemicals and Reagents --- p.53 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Preparation of Primary Cultured Rat Astrocytes --- p.55 / Chapter 2.2.2 --- Measuring Cell Viability: MTT Assay --- p.56 / Chapter 2.2.3 --- Measuring Cell Proliferation: 3H Thymidine Incorporation Assay --- p.57 / Chapter 2.2.4 --- Measuring Glucose Uptake: Zero-trans Glucose Uptake Assay --- p.58 / Chapter 2.2.5 --- Measuring Ascorbic Acid Accumulation --- p.60 / Chapter 2.2.6 --- Total Protein Extraction --- p.61 / Chapter 2.2.7 --- Western Blotting --- p.62 / Chapter 2.2.8 --- Immunostaining --- p.64 / Chapter 2.2.9 --- Isolation of RNA --- p.64 / Chapter 2.2.10 --- Measurement of RNA Yield --- p.65 / Chapter 2.2.11 --- RNA Gel Electrophoresis --- p.66 / Chapter 2.2.12 --- Reverse Transcription (RT) --- p.66 / Chapter 2.2.13 --- Polymerase Chain Reaction (PCR) --- p.67 / Chapter 2.2.14 --- Separation of PCR Products by Agarose Gel Electrophoresis --- p.67 / Chapter 2.2.15 --- Quantization of PCR Products and Western Blotting --- p.68 / Chapter 2.2.16 --- Real-time PCR --- p.68 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Role of Calcium Ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.70 / Chapter 3.1.1 --- Effects of Changes of Extracellular Ca2+ on Astrocyte Viability --- p.72 / Chapter 3.1.2 --- Effects of Other Divalent Ions on Astrocyte Viability --- p.74 / Chapter 3.1.3 --- Effects of Changes of Intracellular Ca2+ on Astrocyte Viability --- p.78 / Chapter 3.1.4 --- Role of Ca2+ on TNF-α-mduced Proliferation in Astrocytes --- p.85 / Chapter 3.1.5 --- Role of Other Divalent Ions on tnf-α-mduced Proliferation in Astrocytes --- p.90 / Chapter 3.2 --- Effect of Cytokines on Glucose Uptake in Rat Astrocytes --- p.95 / Chapter 3.2.1 --- Basal level of Glucose Uptake in Astrocytes and Effects of Cytokines on Glucose Uptake in Astrocytes --- p.95 / Chapter 3.2.2 --- Signaling Cascade of LPS- and TNF-α-induced Glucose Uptake in Astrocytes --- p.120 / Chapter (A) --- TNFR Subtypes Mediating TNF-a-induced Glucose Uptake --- p.121 / Chapter (B) --- MAPK --- p.125 / Chapter (C) --- PKA --- p.133 / Chapter (D) --- NFkB --- p.139 / Chapter (E) --- Other Mechanisms / Signalling molecules --- p.150 / Chapter (1) --- Interaction with β-Adrenegic Mechanism / Chapter (2) --- Role of cGMP --- p.154 / Chapter (3) --- Effect of Mg2+ on LPS- / TNF-α- induced Glucose Uptake in Astrocytes --- p.156 / Chapter (4) --- Possible Involvement of IGF-1 System --- p.160 / Chapter 3.2.3 --- Summary --- p.163 / Chapter 3.3 --- Effects of LPS and Cytokines on AA Accumulation in Astrocytes --- p.164 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Role of Calcium ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.177 / Chapter 4.1.1 --- Drastic Changes in Extracellular Ca2+ Caused Astrocyte Death --- p.178 / Chapter 4.1.2 --- Extraordinary Role of Ca2+ in Astrocytes Survival --- p.178 / Chapter 4.1.3 --- Elevation of [Ca2+]i Reduced Astrocyte Viability --- p.180 / Chapter 4.1.4 --- Failure of Verapamil to Block TNF-α-induced Astrocyte Proliferation --- p.182 / Chapter 4.2 --- Hypothesis for the Relationship between Cytokines and Energy Metabolism --- p.185 / Chapter 4.2.1 --- Mechanism and Signaling Cascade of the Elevated Glucose Uptake --- p.186 / Chapter 4.2.2 --- Increased Glucose Uptake by Cytokines: Friend or Foe? --- p.191 / Chapter 4.2.3 --- Depletion of AA Pool by LPS --- p.194 / Chapter 4.2.4 --- Possible Bedside Application of the Findings --- p.195 / Chapter 4.3 --- Prospects of This Study and Concluding Remarks --- p.197 / Appendix --- p.201 / References --- p.202

Ngan, Kit Shan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 176-187). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Thesis (Ph. D.)--West Virginia University, 2003. / Title from document title page. Document formatted into pages; contains xv, 190 p. : ill. (some col.). Includes abstract. Includes bibliographical references.