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A biochemical study of cell death, apoptosis, in macrophages.

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

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_320668
Date January 1995
ContributorsChan, Yee Man Elaine., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xi, 143 leaves : ill. (some mounted col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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