by Catherine Mary (Kay) Roche. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 195-228). / Abstract / Acknowledgements / Publications / Abbreviations / Chapter Chapter 1 --- Introduction --- p.1-26 / Chapter 1.1 --- Historical Background / General information --- p.2 / Chapter 1.2 --- Mast Cell Mediators --- p.3 / Chapter 1.2.1 --- Pre-formed Mediators --- p.4 / Chapter 1.2.2 --- Newly Synthesised Lipid Mediators --- p.5 / Chapter 1.3 --- Mast Cell Activation --- p.7 / Chapter 1.3.1 --- The Antigenic Pathway of Mast Cell Activation --- p.7 / Chapter 1.3.2 --- The Non-Antigenic Pathway of Mast Cell Activation --- p.15 / Chapter 1.4 --- Phospholipase enzymes --- p.17 / Chapter 1.4.1 --- Phospholipase A2 --- p.18 / Chapter 1.4.2 --- The type I PLA2 --- p.20 / Chapter 1.4.3 --- The type II PLA2 --- p.22 / Chapter 1.5 --- Aims of the Study --- p.26 / Chapter Chapter 2 --- Materials and Materials --- p.27-49 / Chapter 2.1 --- Materials --- p.28 / Chapter 2.2 --- Buffers --- p.30 / Chapter 2.3 --- Secretory stimuli --- p.31 / Chapter 2.4 --- Source of mast cells --- p.32 / Chapter 2.4.1 --- Sensitization of animals --- p.33 / Chapter 2.4.2 --- Isolation of rat peritoneal mast cells --- p.33 / Chapter 2.4.3 --- Purification of rat peritoneal mast cells --- p.34 / Chapter 2.4.4 --- Isolation of human lung mast cells --- p.34 / Chapter 2.4.5 --- Partial purification of human lung mast cells --- p.35 / Chapter 2.4.6 --- Isolation of guinea pig lung mast cells --- p.36 / Chapter 2.5 --- General procedure for studying histamine release from isolated mast cells --- p.36 / Chapter 2.5.1 --- Procedure for investigating the effect of sPLA2 on the spontaneous histamine release from isolated mast cells --- p.37 / Chapter 2.5.2 --- Procedure for investigating the effect of heat treated sPLA2-I on histamine release from isolated RPMC --- p.37 / Chapter 2.5.3 --- Procedure for investigating the effect of antimycin A on histamine release induced by sPLA2-I on isolated RPMC --- p.38 / Chapter 2.5.4 --- Procedure for investigating the effects of extracellular calcium on histamine release induced by sPLA2-I on isolated RPMC --- p.38 / Chapter 2.5.5 --- Procedure for investigating the effects of the sPLA2 enzymes on activated mast cells --- p.39 / Chapter 2.5.6 --- Procedure for investigating the effect of preincubation time on sPLA2 induced histamine release from immunologically activated RPMC --- p.39 / Chapter 2.5.7 --- Procedure for investigating the effects of the PLA2 inhibitors --- p.39 / Chapter 2.5.8 --- Procedure for investigating the effects of the cyclo- oxygenase and lipoxygenase inhibitors on sPLA2 --- p.40 / Chapter 2.6 --- Histamine Assay --- p.41 / Chapter 2.7 --- Measurement of prostaglandin D2 --- p.42 / Chapter 2.7.1 --- Prostaglandin D2 methoxime enzyme immunoassay kit --- p.42 / Chapter 2.7.2 --- The Prostaglandin D2 assay --- p.43 / Chapter 2.7.3 --- PGD2 release from purified rat peritoneal mast cells --- p.43 / Chapter 2.7.4 --- Derivatization of PGD2-MOX --- p.44 / Chapter 2.7.5 --- PGD2 assay procedure --- p.44 / Chapter 2.8 --- Measurement of intracellular calcium --- p.47 / Chapter 2.8.1 --- Fluorescent calcium indicator fura-2 --- p.47 / Chapter 2.8.2 --- Cell loading with the calcium indicator --- p.47 / Chapter 2.8.3 --- Measurement of fura-2 fluorescence --- p.47 / Chapter 2.9 --- Cell counts --- p.49 / Chapter 2.10 --- Data analysis --- p.49 / Chapter Chapter 3 --- Effects of sPLA2 on rat peritoneal mast cells --- p.50-157 / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.2 --- Methodology --- p.52 / Chapter Part 1 --- Effects of sPLA2 on unstimulated rat peritoneal mast cells --- p.52-86 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- Effects of sPLA2-I on the spontaneous histamine release from RPMC --- p.52 / Chapter 3.3.2 --- Time course of sPLA2-I induced histamine release --- p.53 / Chapter 3.3.3 --- Effect of heat treated sPLA2-I on histamine release --- p.53 / Chapter 3.3.4 --- Effects of antimycin A on sPLA2-I induced histamine release --- p.54 / Chapter 3.3.5 --- Effects of extracellular calcium on sPLA2-I induced histamine release --- p.54 / Chapter 3.3.6 --- Effects of PLA2 inhibitors on sPLA2-I induced histamine release --- p.54 / Chapter 3.3.7 --- Effects of sPLA2-I on PGD2 production --- p.56 / Chapter 3.3.8 --- Effects of sPLA2-II on the spontaneous histamine release from RPMC --- p.57 / Chapter 3.3.9 --- Effects of PLA2 inhibitors on sPLA2-II induced histamine release from purified RPMC --- p.57 / Chapter 3.3.10 --- Effects of sPLA2-II on PGD2 production --- p.57 / Chapter 3.4 --- Discussion --- p.75 / Chapter 3.4.1 --- Effects of sPLA2-I on RPMC --- p.75 / Chapter 3.4.2 --- Effects of sPLA2-II on RPMC --- p.83 / Chapter Part 2 --- Effects of sPLA2 on stimulated rat peritoneal mast cells --- p.87-130 / Chapter 3.5 --- Results --- p.87 / Chapter 3.5.1 --- Effects of sPLA2-I on histamine release from immunologically activated RPMC --- p.87 / Chapter 3.5.2 --- Effects of preincubation time on sPLA2-I enhanced histamine release from immunologically activated RPMC --- p.88 / Chapter 3.5.3 --- Effects of p-BPB on sPLA2-I enhanced histamine release from immunologically activated RPMC --- p.88 / Chapter 3.5.4 --- Effects of 12-epi-scalaradial on sPLA2-I enhanced histamine release from immunologically activated RPMC --- p.89 / Chapter 3.5.5 --- Effects of MAFP on sPLA2-I enhanced histamine release from immunologically activated RPMC --- p.89 / Chapter 3.5.6 --- Effects of sPLA2-I on PGD2 production from immunologically activated RPMC --- p.90 / Chapter 3.5.7 --- Effects of flurbiprofen and zileuton on sPLA2-I induced histamine release and enhanced histamine release from RPMC --- p.90 / Chapter 3.5.8 --- Effects of sPLA2-II on histamine release from immunologically activated RPMC --- p.91 / Chapter 3.5.9 --- Effects of preincubation time on sPLA2-II enhanced histamine release from immunologically activated RPMC --- p.92 / Chapter 3.5.10 --- Effects of p-BPB on sPLA2-II enhanced histamine release from immunologically activated RPMC --- p.92 / Chapter 3.5.11 --- Effects of 12-epi-scalaradial on sPLA2-II enhanced histamine release from immunologically activated RPMC --- p.92 / Chapter 3.5.12 --- Effects of MAFP on sPLA2-II enhanced histamine release from immunologically activated RPMC --- p.93 / Chapter 3.5.13 --- Effects of sPLA2-II on PGD2 production from immunologically activated RPMC --- p.93 / Chapter 3.5.14 --- Effects of flurbiprofen and zileuton on sPLA2-II induced histamine release and enhanced histamine release from RPMC --- p.93 / Chapter 3.6 --- Discussion --- p.124 / Chapter 3.6.1 --- Effects of sPLA2 on immunologically induced histamine release from activated RPMC --- p.124 / Chapter 3.6.2 --- Effects of sPLA2 on PGD2 production from immunologically activated RPMC --- p.128 / Chapter 3.6.3 --- Effects of flurbiprofen and zileuton on sPLA2 induced and enhanced histamine release from RPMC --- p.129 / Chapter Part 3 --- Investigation of the mode of action(s) of sPLA2 on RPMC --- p.131-157 / Chapter 3.7 --- Results --- p.131 / Chapter 3.7.1 --- Effects of sPLA2-I on non-immunologically activated RPMC --- p.131 / Chapter 3.7.2 --- Effects of sPLA2-II on non-immunologically activated RPMC --- p.131 / Chapter 3.7.3 --- Effects of sPLA2-I on [Ca2+]i in immunologically activated RPMC --- p.132 / Chapter 3.7.4 --- Effects of sPLA2-II on [Ca2+]i in immunologically activated RPMC --- p.133 / Chapter 3.8 --- Discussion --- p.145 / Chapter 3.9 --- Summary of the possible mechanism (s) of action of sPLA2 --- p.149 / Chapter 3.10 --- Conclusions --- p.156 / Chapter 3.11 --- Future studies --- p.157 / Chapter Chapter 4 --- Effects of sPLA2 on human and guinea pig lung mast cells --- p.158-194 / Chapter 4.1 --- Introduction --- p.159 / Chapter 4.2 --- Methodology --- p.162 / Chapter 4.3 --- Results --- p.163 / Chapter 4.3.1 --- Effects of sPLA2 on the spontaneous histamine release from HLMC --- p.163 / Chapter 4.3.2 --- Effects of sPLA2 on immunologically induced histamine release from HLMC --- p.163 / Chapter 4.3.3 --- Effects of sPLA2 on A23187 induced histamine release from HLMC --- p.164 / Chapter 4.3.4 --- Effects of p-BPB on the inhibitory activity of sPLA2-I --- p.165 / Chapter 4.3.5 --- Effects of sPLA2 on partially purified HLMC --- p.165 / Chapter 4.3.6 --- Effects of sPLA2 on the spontaneous histamine release from GPLMC --- p.166 / Chapter 4.3.7 --- Effects of sPLA2 on immunologically induced histamine release from GPLMC --- p.167 / Chapter 4.4 --- Discussion --- p.189 / Chapter 4.5 --- Conclusions --- p.194 / References --- p.195-228
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_321691 |
| Date | January 1996 |
| Contributors | Roche, Catherine Mary., Chinese University of Hong Kong Graduate School. Division of Basic Medical Sciences. |
| Publisher | Chinese University of Hong Kong |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, 228 leaves : ill. ; 30 cm. |
| Rights | Use 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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