by Chan Yee Man Elaine. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 130-143). / Abstract --- p.I / Acknowledgements --- p.III / Abbreviations --- p.IV / Objectives of the study --- p.VII / Table of Contents --- p.VIII / Chapter Section 1 --- Introduction --- p.1 / Chapter 1.1 --- Necrosis vs Apoptosis --- p.2 / Chapter 1.2 --- Cell Death by Apoptosis --- p.4 / Chapter 1.3 --- The Biochemistry of Nitric Oxide --- p.9 / Chapter 1.4 --- Mechanisms of NO Action --- p.14 / Chapter 1.5 --- Signal Transduction Pathways to Apoptosis --- p.17 / Chapter 1.5.1 --- Regulation by Ca2+ --- p.17 / Chapter 1.5.2 --- Protein Kinase C --- p.20 / Chapter 1.5.3 --- cAMP --- p.21 / Chapter 1.5.4 --- Protein tyrosine kinase --- p.21 / Chapter 1.5.5 --- Ceramide --- p.22 / Chapter 1.5.6 --- pH --- p.23 / Chapter 1.5.7 --- Oxygen Radicals --- p.23 / Chapter 1.5.8 --- Anchorage Dependence and Extracellular Matrix --- p.24 / Chapter Section 2 --- Materials and Methods --- p.27 / Chapter 2.1 --- Materials --- p.28 / Chapter 2.1.1 --- Animal --- p.28 / Chapter 2.1.2 --- Cell line --- p.28 / Chapter 2.1.3 --- "Culture media, buffers and chemicals" --- p.28 / Chapter 2.1.4 --- Dye solutions --- p.30 / Chapter 2.1.5 --- Reagents and buffers for polyacrylamide gel electrophoresis (PAGE) --- p.31 / Chapter 2.1.6 --- Reagents and buffers for Western blotting --- p.32 / Chapter 2.1.7 --- Reagents and buffers for agarose gel electrophoresis --- p.33 / Chapter 2.2 --- Methods --- p.35 / Chapter 2.2.1 --- Cell culture --- p.35 / Chapter 2.2.2 --- [3H]-Thymidine incorporation --- p.35 / Chapter 2.2.3 --- MTT assay --- p.36 / Chapter 2.2.4 --- Determination of NO by Griess assay --- p.36 / Chapter 2.2.5 --- Observation of apoptotic morphology of cells by confocal laser scanning microscopy (CLSM) --- p.37 / Chapter 2.2.6 --- Determination of cell death induced by NO producing drugs --- p.38 / Chapter 2.2.7 --- Determination of cell death induced by Concanavalin A --- p.38 / Chapter 2.2.8 --- Determination of effect of nitric oxide synthase (NOS) inhibitor on cell death induced by Con A --- p.39 / Chapter 2.2.9 --- Determination of the requirement of Ca2+ in cell death induced by NO producing drugs --- p.39 / Chapter 2.2.10 --- Determination of the requirement of cGMP in cell death induced by NO producing drugs --- p.40 / Chapter 2.2.11 --- Determination of cell death induced by PKC activation and depletion --- p.40 / Chapter 2.2.12 --- Determination of effect of PKC depletion on cell death induced by NO producing drugs and Con A --- p.41 / Chapter 2.2.13 --- Observation of immunofluorescence by confocal laser scanning microscopy --- p.41 / Chapter 2.2.14 --- Preparation of protein samples for PAGE --- p.42 / Chapter 2.2.15 --- Polyacrylamide gel electrophoresis --- p.43 / Chapter 2.2.16 --- Western blotting of PKC --- p.44 / Chapter 2.2.17 --- Preparation of DNA samples from cells --- p.46 / Chapter 2.2.18 --- Agarose gel electrophoresis of DNA --- p.47 / Chapter 2.2.19 --- Statistical analysis --- p.48 / Chapter Section 3 --- Results --- p.49 / Chapter 3.1 --- Induction of apoptosis in macrophages by NO producing drugs --- p.50 / Chapter 3.2 --- Apoptosis induced by NO producing drugs was caused by NO --- p.54 / Chapter 3.3 --- Signal transduction pathways to the NO-induced cell death in macrophages --- p.66 / Chapter 3.3.1 --- Calcium ion --- p.66 / Chapter 3.3.2 --- cGMP --- p.68 / Chapter 3.3.3 --- Protein kinase C --- p.71 / Chapter 3.4 --- Induction of apoptosis by Con A --- p.89 / Chapter 3.5 --- Tubulin structure in Con A-treated cells --- p.94 / Chapter 3.6 --- Nitric oxide and Con A-induced cell death in macrophages --- p.96 / Chapter 3.7 --- Effect ofNOS inhibitor on cell death induced by Con A --- p.102 / Chapter 3.8 --- Involvement of PKC in the Con A-induced cell death in macrophages --- p.106 / Chapter Section 4 --- Discussion --- p.114 / Chapter 4.1 --- Induction of apoptosis in macrophages by NO producing drugs --- p.118 / Chapter 4.2 --- Signal transduction pathways to the NO-induced cell death in macrophages --- p.120 / Chapter 4.2.1 --- Ca2+ ion --- p.120 / Chapter 4.2.2 --- cGMP --- p.120 / Chapter 4.2.3 --- Protein kinase C --- p.121 / Chapter 4.3 --- Induction of apoptosis by Con A --- p.124 / Chapter 4.4 --- Tubulin structure in Con A-treated cells --- p.124 / Chapter 4.5 --- Nitric oxide and Con A-induced cell death --- p.125 / Chapter 4.6 --- Effect ofNOS inhibitor on cell death induced by Con A --- p.125 / Chapter 4.7 --- Involvement of PKC in the Con A-induced cell death in macrophages --- p.127 / Chapter Section 5 --- Bibliography --- p.129 / References --- p.130

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