by Yuen Mei Ling, Miranda. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1989. / Bibliography: leaves 73-83.

Wai Kei Chan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 155-164). / Abstracts in English and Chinese. / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- Sulfonylureas --- p.1 / Chapter 1.2. --- Biological action of sulfonylurea drugs --- p.2 / Chapter 1.2.1 --- Effects on pancreatic β cells --- p.5 / Chapter 1.2.2. --- Effects on cardiac myocytes --- p.7 / Chapter 1.2.3. --- Effects on smooth muscle cells --- p.11 / Chapter 1.2.4. --- Effects on endothelial cells --- p.14 / Chapter 1.3. --- Side effects and toxicity --- p.15 / Chapter 1.4. --- Objectives of the present study --- p.17 / Chapter Chapter 2 --- Methods and Marterials / Chapter 2.1. --- Tissue and Cell Preparation --- p.20 / Chapter 2.1.1. --- Preparation of the isolated rat aorta and mesenteric artery --- p.20 / Chapter 2.1.2. --- Removal of the functional endothelium --- p.20 / Chapter 2.1.3. --- Cell culture --- p.21 / Chapter 2.1.3.1. --- Materials --- p.21 / Chapter 2.1.3.2. --- Aortic smooth muscle cells in primary culture --- p.21 / Chapter 2.1.3.3. --- Aortic endothelial cells in primary culture --- p.23 / Chapter 2.1.3.4. --- Cultured rat aortic smooth muscle cell line (A7r5) --- p.23 / Chapter 2.1.3.5. --- Cultured human umbilical vein endothelial cell line (ECV-304) --- p.24 / Chapter 2.1.3.6. --- Cell subculture --- p.24 / Chapter 2.1.3.7. --- Immunostaining of endothelial cells in primary culture --- p.24 / Chapter 2.2. --- Organ Bath Set-up --- p.25 / Chapter 2.3. --- Force Measurement --- p.28 / Chapter 2.3.1. --- Vascular action of glibenclamide --- p.28 / Chapter 2.3.1.1. --- Antagonistic effect of glibenclamide on relaxation induced by K+ channel openers --- p.28 / Chapter 2.3.1.2. --- Relaxant response of glibenclamide --- p.29 / Chapter 2.3.1.3. --- Role of endothelium-derived vasoactive factors in glibenclamide induced relaxation --- p.29 / Chapter 2.3.1.4. --- Effect of endothelial prostanoids in glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.5. --- Effects of putative K+ channel blockers on glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.6. --- Effect of glibenclamide on high K+- and CaCl2-induced contraction --- p.31 / Chapter 2.3.1.7. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.32 / Chapter 2.3.1.8. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.32 / Chapter 2.3.2. --- Vascular action of glipizide --- p.33 / Chapter 2.3.3. --- Vascular action of tolbutamide --- p.33 / Chapter 2.3.3.1. --- Contractile response of tolbutamide --- p.33 / Chapter 2.3.3.2. --- Effects of inhibitors of endothelium-derived factors --- p.33 / Chapter 2.3.3.3. --- Effects of inhibitors of Ca2+ influx --- p.34 / Chapter 2.3.3.4. --- Effect of protein kinase C inhibitor --- p.34 / Chapter 2.3.3.5. --- Effects of neural factors --- p.34 / Chapter 2.4. --- Cyclic GMP measurement --- p.35 / Chapter 2.4.1. --- Material --- p.35 / Chapter 2.4.2. --- Methods --- p.35 / Chapter 2.4.2.1. --- Tissue preparation --- p.35 / Chapter 2.4.2.2. --- Plasma and tissue according to protocols provided by Amersham --- p.35 / Chapter 2.4.2.3. --- Cyclic GMP content measurement --- p.36 / Chapter 2.4.2.4. --- Protein content measurement --- p.39 / Chapter 2.4.2.5. --- Cyclic GMP measurement protocol --- p.40 / Chapter 2.5. --- Ca2+ measurement --- p.40 / Chapter 2.5.1. --- Materials --- p.40 / Chapter 2.5.1.1. --- PTI RatioMaster Fluorescence System --- p.40 / Chapter 2.5.1.2. --- Confocal Imaging System --- p.42 / Chapter 2.5.2. --- Method --- p.42 / Chapter 2.5.3. --- Protocols for Ca2+ measurement --- p.45 / Chapter 2.5.3.1. --- Effect of glibenclamide in endothelial cells --- p.45 / Chapter 2.5.3.2. --- Effect of glibenclamide in vascular smooth muscle cells --- p.45 / Chapter 2.5.3.3. --- Effect of tolbutamide in vascular smooth muscle cells --- p.46 / Chapter 2.6. --- Cell proliferation --- p.45 / Chapter 2.6.1. --- Materials --- p.45 / Chapter 2.6.2. --- Method --- p.46 / Chapter 2.6.3. --- Protocols for cell proliferation --- p.47 / Chapter 2.6.3.1. --- Effect of glibenclamide on endothelial cell proliferation --- p.47 / Chapter 2.6.3.2. --- Effect of glibenclamide on aortic smooth muscle cell proliferation --- p.47 / Chapter 2.7. --- Chemicals and solutions --- p.48 / Chapter 2.8. --- Statistical analysis --- p.50 / Chapter Chapter 3 --- Results / Chapter 3.1. --- Glibenclamide --- p.51 / Chapter 3.1.1. --- Effect of glibenclamide on the K+ channel activity --- p.51 / Chapter 3.1.2. --- Relaxant response of glibenclamide --- p.55 / Chapter 3.1.3. --- Effects of inhibitors of nitric oxide activity on glibenclamide- induced relaxation --- p.57 / Chapter 3.1.4. --- Role of endothelial relaxing prostanoids in glibenclamide-induced relaxation --- p.69 / Chapter 3.1.5. --- Effect of putative K+ channel blockers on glibenclamide-induced relaxation --- p.73 / Chapter 3.1.6. --- Effect of glibenclamide on high K+-induced arterial contraction --- p.75 / Chapter 3.1.7. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.83 / Chapter 3.1.8. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.83 / Chapter 3.2 --- Glipizide --- p.85 / Chapter 3.2.1. --- Relaxant response of glipizide --- p.85 / Chapter 3.3. --- Tolbutamide --- p.91 / Chapter 3.3.1. --- Contractile response to tolbutamide --- p.91 / Chapter 3.3.2. --- Effects of endothelium-derived factors --- p.94 / Chapter 3.3.3. --- Effects of inhibitors of Ca2+ influx on tolbutamide-induced contraction --- p.98 / Chapter 3.3.4. --- "Effects of forskolin, sodium nitroprusside, staurosporine on tolbutamide-induced contraction" --- p.102 / Chapter 3.3.5. --- Effect of neural factors --- p.106 / Chapter 3.4. --- Effect of glibenclamide on cGMP levels --- p.112 / Chapter 3.5. --- Effect of glibenclamide on intracellular[Ca2+ ] in cultured endothelial cells --- p.112 / Chapter 3.6. --- Effect of glibenclamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.115 / Chapter 3.7. --- Effect of tolbutamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.121 / Chapter 3.8. --- Effect of glibenclamide on proliferation of cultured endothelial cells --- p.121 / Chapter 3.9. --- Effect of glibenclamide on proliferation of cultured aortic smooth muscle cells --- p.123 / Chapter Chapter 4 --- Discussion / Chapter 4.1. --- Effect of glibenclamide --- p.133 / Chapter 4.2. --- Effect of glipizide --- p.143 / Chapter 4.3. --- Effect of tolbutamide --- p.144 / Chapter 4.4. --- Conclusion --- p.152 / References --- p.155 / Publications --- p.163

Sha Ou. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 167-189). / Abstracts in English and Chinese. / abstract --- p.i / 中文摘要 --- p.iv / acknowledgements --- p.vii / Chapter chapter 1. --- introduction --- p.1 / Chapter 1.1 --- Overview --- p.1 / Chapter 1.2 --- Ribosome-inactivating proteins (RIPs) --- p.1 / Chapter 1.2.1 --- Classification --- p.2 / Chapter 1.2.2 --- Structure --- p.3 / Chapter 1.2.3 --- Enzymatic activities --- p.3 / Chapter 1.3 --- Type II RIPs --- p.5 / Chapter 1.3.1 --- Ricin --- p.5 / Chapter 1.3.2 --- Ricinus communis agglutinin (RCA) --- p.6 / Chapter 1.3.3 --- Intracellular mechanism --- p.7 / Chapter 1.3.4 --- Application of RIPs in neuroscience research: suicide axonal transport --- p.10 / Chapter 1.4 --- Type I RIPs --- p.12 / Chapter 1.4.1 --- Trichosanthin (TCS) --- p.12 / Chapter 1.4.2 --- Ricin A chain (RTA) --- p.15 / Chapter 1.4.3 --- Medical applications: immunolesioning and immunotherapy --- p.16 / Chapter 1.5 --- The types of Cell death --- p.17 / Chapter 1.5.1 --- Necrosis --- p.18 / Chapter 1.5.2 --- Apoptosis --- p.18 / Chapter 1.6 --- Inflammations --- p.21 / Chapter 1.6.1 --- Acute inflammation --- p.21 / Chapter 1.6.2 --- Chronic inflammation --- p.22 / Chapter 1.6.3 --- Retinitis --- p.22 / Chapter 1.7 --- Eye model for neurotoxicity studies in CNS --- p.23 / Chapter 1.8 --- Objective of present study --- p.24 / Chapter CHAPTER 2. --- MATERIALS AND METHODS --- p.25 / Chapter 2.1 --- Plan of this chapter --- p.25 / Chapter 2.2 --- Toxins and methods used --- p.25 / Chapter 2.3 --- Animals --- p.26 / Chapter 2.4 --- Preparation of toxin solutions --- p.27 / Chapter 2.4.1 --- RIP solutions --- p.27 / Chapter 2.4.2 --- Labeling type I RIPs with fluorescence --- p.27 / Chapter 2.4.3 --- Control solutions --- p.29 / Chapter 2.5 --- Administrations of solutions --- p.30 / Chapter 2.5.1 --- Basic procedures of vitreous chamber injection --- p.30 / Chapter 2.5.2. --- Injection of trichosanthin (TCS) --- p.31 / Chapter 2.5.3 --- Injection of ricin A chain (RTA) --- p.31 / Chapter 2.5.4 --- Injection of ricinus communis agglutinin (RCA) --- p.32 / Chapter 2.5.5 --- Administration of FITC-TCS --- p.33 / Chapter 2.5.6 --- Administration of FITC-RTA --- p.33 / Chapter 2.6 --- Retinal tissue processing --- p.33 / Chapter 2.6.1 --- Paraffin method --- p.34 / Chapter 2.6.2 --- Cryostatic method --- p.35 / Chapter 2.6.3 --- Electron microscopic method --- p.35 / Chapter 2.7 --- General effects of RIPs on rat retinas --- p.36 / Chapter 2.7.1 --- Hematoxylin-and-eosin staining --- p.36 / Chapter 2.7.2 --- Retinal thickness --- p.37 / Chapter 2.7.3 --- Pathological changes --- p.38 / Chapter 2.7.4 --- Dosage study on TCS --- p.39 / Chapter 2.7.5 --- Statistics --- p.40 / Chapter 2.8 --- Mechanisms of cell death --- p.40 / Chapter 2.8.1 --- Terminal dUTP nick-end labeling (TUNEL) --- p.40 / Chapter 2.8.2 --- Immunohistochemistry for caspase-3 --- p.42 / Chapter 2.8.3 --- Double staining of cleaved caspase-3 and TUNEL --- p.42 / Chapter 2.8.4 --- Electronic microscope observation --- p.43 / Chapter 2.9 --- Entry of type I RIPs into cells --- p.43 / Chapter 2.9.1 --- Propidium iodide staining --- p.43 / Chapter 2.9.2 --- Immunohistochemical localization of Muller cells --- p.44 / Chapter 2.9.3 --- Double staining of Muller cells and TUNEL --- p.44 / Chapter 2.9.4 --- Confocal microscope --- p.44 / Chapter 2.10 --- Reactions of glial cells --- p.45 / Chapter CHAPTER 3. --- RESULTS --- p.47 / Chapter 3.1 --- Preparation of fluorescein-type I RIP conjugates --- p.47 / Chapter 3.1.1 --- Conjugate of FITC-TCS --- p.47 / Chapter 3.1.2 --- Conjugate of FITC-RTA --- p.47 / Chapter 3.2 --- Effects of TCS on retina --- p.47 / Chapter 3.2.1 --- Retina cell count - a dose-dependence study --- p.48 / Chapter 3.2.2 --- Retinal thickness measurement - a time-course study --- p.49 / Chapter 3.2.3 --- Pathological changes --- p.50 / Chapter 3.3 --- Effects of RTA on retina --- p.51 / Chapter 3.3.1 --- Retinal thickness measurement - a time-course study --- p.51 / Chapter 3.3.2 --- Pathological changes --- p.53 / Chapter 3.4 --- Effects of RCA on retina --- p.54 / Chapter 3.4.1 --- Retinal thickness measurement --- p.54 / Chapter 3.4.2 --- Pathological changes --- p.55 / Chapter 3.5 --- Summary of results: general effects of RIPs --- p.56 / Chapter 3.6 --- Cell death - TUNEL method --- p.56 / Chapter 3.6.1 --- TCS experiment --- p.57 / Chapter 3.6.2 --- RTA experiment --- p.58 / Chapter 3.6.3 --- RCA experiment --- p.58 / Chapter 3.7 --- Cell death 一 cleaved caspase-3 immunohistochemistry --- p.58 / Chapter 3.7.1 --- TCS experiment --- p.59 / Chapter 3.7.2 --- RTA experiment --- p.59 / Chapter 3.8 --- EM observation --- p.59 / Chapter 3.8.1 --- TCS experiment --- p.59 / Chapter 3.8.2 --- RTA experiment --- p.60 / Chapter 3.9 --- Summary of results: mode of cell death --- p.60 / Chapter 3.10 --- Localisation of type I RIPs --- p.61 / Chapter 3.10.1 --- FITC-TCS --- p.62 / Chapter 3.10.2 --- FITC-TCS and Muller cell double staining --- p.63 / Chapter 3.10.3 --- Muller cell and TUNEL double staining --- p.64 / Chapter 3.10.4 --- FITC-RTA --- p.64 / Chapter 3.10.5 --- Summary of results: route of intoxication --- p.65 / Chapter 3.11 --- Glial cell reactions after RIP treatment --- p.65 / Chapter 3.11.1 --- TCS experiment --- p.65 / Chapter 3.11.2 --- RTA experiment --- p.66 / Chapter 3.11.3 --- RCA experiment --- p.67 / Chapter 3.11.4 --- Summary of results: glial reactions --- p.67 / Chapter CHAPTER 4. --- DISCUSSION --- p.69 / Chapter 4.1 --- General effects of RIPs on rat retinas --- p.69 / Chapter 4.1.1 --- Effects of trichosanthin (TCS) --- p.69 / Chapter 4.1.2 --- Effects of ricin A chain (RTA) --- p.71 / Chapter 4.1.3 --- Effects of ricinus communis agglutinin (RCA) --- p.73 / Chapter 4.2 --- The mechanisms of cell death --- p.74 / Chapter 4.2.1 --- Cell death caused by TCS --- p.75 / Chapter 4.2.2 --- Caspase-3 and the retina of RCS rat --- p.77 / Chapter 4.2.3 --- Cell death caused by RTA --- p.78 / Chapter 4.2.4 --- Cell death caused by RCA --- p.80 / Chapter 4.2.5 --- Mechanism of RTA - induced necrosis --- p.81 / Chapter 4.3 --- The mechanisms of type I RIPs entering cells --- p.82 / Chapter 4.3.1 --- Transport of TCS in retinal cells --- p.82 / Chapter 4.3.2 --- The uptake of Pure FITC by rat retina --- p.85 / Chapter 4.4 --- Reactions of glial cells --- p.85 / Chapter 4.4.1 --- Glial cell reactions in TCS experiment --- p.86 / Chapter 4.4.2 --- Glial cell reactions in RTA and RCA experiments --- p.87 / Chapter 4.5 --- Possible applications of RIPs on retinal studies --- p.88 / Chapter 4.5.1 --- Potential applications of TCS --- p.88 / Chapter 4.5.2 --- Possible uses of RTA and RCA --- p.90 / Chapter CHAPTER 5. --- CONCLUSIONS --- p.91 / "FIGURES, TABLES, GRAPHS, AND LEGENDS" --- p.93 / APPENDICES --- p.154 / Appendix A Source of materials --- p.154 / Appendix B Dosages for vitreous chamber injection --- p.156 / Appendix C Protocol of conjugate fluorescein to proteins --- p.157 / Appendix D Electronic Microscope methods --- p.160 / Appendix E Histological methods --- p.162 / Appendix F Protocols of TUNEL --- p.163 / Appendix G Protocols of Immunohistochemistry staining --- p.165 / REFERENCES --- p.167

by Jing Xian Li. / Thesis (Ph.D.)--Chinese university of Hong Kong, 1998. / Includes bibliographical references (p. 137-151). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Yang, Chao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 137-146). / Abstracts in English and Chinese. / Thesis Committee --- p.i / Acknowledgements --- p.ii / Contents --- p.iii / Declaration --- p.vii / Abstract --- p.viii / 摘要 --- p.xi / Abbreviations --- p.xiii / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- Occurrence and Structure of Phytosterols in Plants --- p.P.1 / Chapter 1.2 --- Biological Effects of Phytosterols / Chapter 1.2.1 --- Cholesterol-lowering Effect of Phytosterols --- p.P.3 / Chapter 1.2.2 --- Anti-cancer Effect of Phytosterols --- p.P.5 / Chapter 1.2.3 --- Anti-proliferative Effect of Phytosterols --- p.P.5 / Chapter 1.3 --- Intake and Absorption of Phytosterols in Human Beings --- p.P.6 / Chapter 1.4 --- Occurrence and Physiological Levels of Phytosterol Oxidation Products (POPs) / Chapter 1.4.1 --- Occurrence of POPs --- p.P.8 / Chapter 1.4.2 --- Physiological Levels of POPs --- p.P.8 / Chapter 1.5 --- Endothelium and the Vascular Tone / Chapter 1.5.1 --- Role of Endothelium in the Control of Vascular Tone --- p.P.11 / Chapter 1.5.2 --- "Endothelial Dysfunction, Cholesterol Oxidation Products (COPs) and Phytosterol Oxidation Products (POPs)" --- p.P.12 / Chapter 1.6 --- Calcium Homeostasis in the Vascular Smooth Muscle Cells (VSMCs) / Chapter 1.6.1 --- Modes of Ca2+ Entry in VSMCs --- p.P.15 / Chapter 1.6.2 --- Modes of Ca2+ Efflux in VSMCs --- p.P.18 / Chapter 1.7 --- Objectives of the Study --- p.P.19 / Chapter CHAPTER TWO: --- β-SITOSTEROL OXIDATION PRODUCTS ATTENUATE VASORELAXATION BY INCREASING REACTIVE OXYGEN SPECIES AND CYCLOOXYGENASE-2 / Chapter 2.1 --- Introduction --- p.P.21 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Preparation of SOPs --- p.P.24 / Chapter 2.2.2 --- Gas Chromatography -mass Spectrometry (GC-MS) Identification of SOPs --- p.P.24 / Chapter 2.2.3 --- Analysis of SOPs --- p.P.25 / Chapter 2.2.4 --- Vessel Preparation --- p.P.25 / Chapter 2.2.5 --- Isometric Force Measurement --- p.P.26 / Chapter 2.2.6 --- Western Blotting --- p.P.27 / Chapter 2.2.7 --- Primary Culture of Rat Aortic Endothelial Cell --- p.P.28 / Chapter 2.2.8 --- Measurement of SOPs-induced Intracellular Oxidative Stress --- p.P.29 / Chapter 2.2.9 --- Drugs --- p.P.30 / Chapter 2.2.10 --- Data Analysis --- p.P.30 / Chapter 2.3 --- Results / Chapter 2.3.1 --- GC-MS Identification of SOPs --- p.P.32 / Chapter 2.3.2 --- Analysis of SOPs --- p.P.34 / Chapter 2.3.3 --- SOPs But Not β-Sitosterol Impaired ACh- and A23187-induced relaxations --- p.P.36 / Chapter 2.3.4 --- Inhibition of COX Pathway Reversed SOPs-induced Impairment in Relaxation --- p.P.39 / Chapter 2.3.5 --- SOPs Elevated Endothelial COX-2 Expression --- p.P.42 / Chapter 2.3.6 --- SOPs Increased COX-2 Expression via An Oxidative Stress-sensitive Pathway --- p.P.45 / Chapter 2.4 --- Discussion --- p.P.52 / Chapter 2.5 --- Conclusion --- p.P.56 / Chapter CHAPTER THREE: --- β-SITOSTEROL OXIDATION PRODUCTS POSSESS POTENTIAL VOCC BLOCKING EFFECT IN VSMCs / Chapter 3.1 --- Introduction / Chapter 3.1.1 --- 2+ Modes of Ca Entry and Efflux in Vascular Smooth Muscle Cells (VSMCs) --- p.P.57 / Chapter 3.1.2 --- Effect of Cholesterol and COPs on VSMCs --- p.P.57 / Chapter 3.2 --- Methodology and Materials / Chapter 3.2.1 --- Vessel Preparation --- p.P.59 / Chapter 3.2.2 --- Isometric Force Measurement iv --- p.P.59 / Chapter 3.2.3 --- Drugs --- p.P.60 / Chapter 3.2.4 --- Data Analysis --- p.P.61 / Chapter 3.3 --- Results / Chapter 3.3.1 --- SOPs but not β-Sitosterol Induced Relaxation in 60 mM K+ -preconstricted Endothelium-denuded Aorta --- p.P.62 / Chapter 3.3.2 --- Both SOPs and β-Sitosterol did not Relax U46619-preconstricted Endothelium-denuded Aorta --- p.P.64 / Chapter 3.3.3 --- Both SOPs and β-Sitosterol did not Relax PDA -preconstricted Endothelium-denuded Aorta --- p.P.66 / Chapter 3.3.4 --- SOPs Attenuated 60 mM K+-induced Contraction --- p.P.68 / Chapter 3.3.5 --- SOPs Attenuated Phenylephrine-induced Contraction --- p.P.70 / Chapter 3.3.6 --- Effect of SOPs on Concentration-dependent Responses to U46619 --- p.P.72 / Chapter 3.3.7 --- Preincubation with Bay K 8644 Abolished SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.74 / Chapter 3.3.8 --- Preincubation with Thapsigargin did not Affect SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.76 / Chapter 3.3.9 --- Preincubation with Ouabain did not Affect SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.78 / Chapter 3.3.10 --- Preincubation with Nickel Potentiated SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.80 / Chapter 3.4 --- Discussion --- p.P.84 / Chapter 3.5 --- Conclusion and Future Work --- p.P.88 / Chapter CHAPTER FOUR: --- INVOLEMENT OF NITRIC OXIDE IN THE PROTECTIVE EFFECTS OF PHYTOSTEROLS AGAINST HOMOCYSTEINE-INDUCED IMPAIRMENT OF ENDOTHELIUM-DEPENDENT RELAXATIONS OF RAT AORTA / Chapter 4.1 --- Introduction --- p.P.89 / Chapter 4.2 --- Materials and Method / Chapter 4.2.1 --- Vessel Preparation --- p.P.93 / Chapter 4.2.2 --- Isometric Force Measurement --- p.P.93 / Chapter 4.2.3 --- Western Blotting --- p.P.94 / Chapter 4.2.4 --- "1,1 -diphenyl-2-picrylhydrazyl (DPPH) Radical Scavenging Capacity" --- p.P.96 / Chapter 4.2.5 --- Primary Culture of Rat Aortic Endothelial Cells V --- p.P.96 / Chapter 4.2.6 --- Measurement Intracellular Oxidative Stress --- p.P.97 / Chapter 4.2.7 --- Nitric Oxide (NO) Measurement --- p.P.97 / Chapter 4.2.8 --- Drugs --- p.P.98 / Chapter 4.2.9 --- Data Analysis --- p.P.99 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Impairment of Endothelium-dependent Relaxation by HC was Reversed by ROS Scavenger --- p.P.100 / Chapter 4.3.2 --- Brassicasterol Reversed HC-induced Endothelial Dysfunction In a Dose-dependent Manner --- p.P.102 / Chapter 4.3.3 --- β-Sitosterol and Stigmasterol Reversed HC-induced Endothelial Dysfunction --- p.P.104 / Chapter 4.3.4 --- Effects of β-Sitosterol Oxidation Products (SOPs) on HC-induced Endothelial Dysfunction --- p.P.106 / Chapter 4.3.5 --- Effects of Brassicasterol and β-Sitosterol on H2O2-induced Impairment of Endothelium-dependent Relaxation --- p.P.108 / Chapter 4.3.6 --- Phytosterols did not Directly Scavenge Free Radicals --- p.P.110 / Chapter 4.3.7 --- "HC and Brassicasterol did not Affect the Expression of SOD-1, SOD-2, eNOS, COX-1 and COX-2 in Aorta" --- p.P.112 / Chapter 4.3.8 --- HC Increased ROS Production in Primary Rat Aortic Endotelial Cells --- p.P.116 / Chapter 4.3.9 --- Brassicasterol did not Reverse the ROS Production by HC treatment In the Endothelial Cells --- p.P.120 / Chapter 4.3.10 --- Effect of L-NAME on Reversing the Effect of Brassicasterol on ACh-induced Relaxation --- p.P.123 / Chapter 4.3.11 --- Brassicasterol Reversed the Inhibitory Effect of HC on ACh-induced NO Production in Endothelial Cells --- p.P.125 / Chapter 4.4 --- Discussion --- p.P.128 / Chapter 4.5 --- Conclusion and Future Work --- p.P.132 / Chapter CHAPTER FIVE: --- CONCLUSIONS AND FUTURE WORK --- p.P.134 / Chapter CHAPTER SIX: --- REFERENCES --- p.P.137

The place of estrogen in women's health remains controversial. Premenopausal women have a lower prevalence of cardiovascular disease (CVD) than men and in observational studies hormone replacement therapy (HRT) decreases CVD in postmenopausal women. However, prospective randomized trials of secondary and primary prevention have failed to substantiate an overall protective effect from HRT and have even shown some harm. To explain this paradox it is necessary to better understand the effects of estrogen on the vascular wall. Estrogen rapidly mediates the activation of eNOS and increases the production of nitric oxide (NO), an important factor for endothelial health. In ovariectomized rats estrogen reduces production of superoxide (O2-) by NAD(P)H oxidase. The decreased function is associated with a decrease in the p47phox component of NAD(P)H oxidase and its interaction with the multicomponent enzyme. In these rats estrogen did not alter eNOS expression and bioavailability of NO, which is in contrast to its acute effects. This highlights the difference between chronic and acute studies. The decrease in O2-production suggests the intracellular signaling. / Estrogen has antiapoptotic effects. Oxidized low-density lipoprotein (oxLDL) and the inflammatory cytokine TNFalpha increased apoptosis which is associated with atherosclerosis. In human umbilical vein endothelial cells (HUVEC), estrogen decreased the extent of TNFalpha and oxLDL induced apoptosis as indicated by the expression of cleaved caspase-3 and FACS assay. Estrogen also preserves the antiapoptotic mitochondrial Bcl-2 and Bcl-xL proteins. / Estrogen has angiogenic properties that can help a healthy endothelium respond to injury. However, estrogen increases the angiogenesis caused by TNFalpha and this could lead to revascularization in the plaques of women with advanced disease. / Overall the balance between the positive and negative aspects of the effects of estrogen on the vascular wall could explain the paradoxical response in older women.

Adult rats produce two main types of ultrasonic vocalizations (USVs), occurring at 22 and 50 kHz USVs. These calls are associated with aversive and rewarding stimuli, respectively. The neural mechanism of amphetamine-induced calling was examined in lesion and antagonist studies. We also tested whether amphetamine-induced 50 kHz USVs could predict individual differences in intravenous self-administration or conditioned place preference behavior. Further experiments examined whether 50 kHz USVs could be evoked by amphetamine-conditioned sensory stimuli and by rewarding electrical brain stimulation. Overall, our experimental findings: (1) identify certain experimental conditions that increase amphetamine-induced 50 kHz calling, (2) provide evidence that these calls may be dependent on mesolimbic dopaminergic transmission, (3) relate individual differences in 50 kHz vocalizing to other behavioural measures of drug reward, and (4) show that in some situations, 50 kHz calls reflect anticipation of expected rewards.

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 4 uppsatser.

Diazoxide, a non-diuretic benzothiadiazlne antihypertensive agent, is thought to act directly upon the vascular smooth muscle of the resistance vessels to exert its therapeutic effects in hypertension. Diazoxide may exert its antihypertensive action by antagonizing calcium in vascular smooth muscle. Wohl et al. (1967 and 1968) have suggested such an interaction based on experiments conducted with isolated rabbit aortae. The present experiments were designed to investigate the possible cellular locus of the postulated interaction of diazoxide with calcium using the isolated anterior mesenteric vein of the rabbit as a model of vascular smooth muscle. This vein is spontaneously motile and possesses characteristics similar to those observed for vessels of the microcirculation. Diazoxide at 10ˉ⁴ M inhibited spontaneous motility and its associated membrane electrical activity, and caused hyperpolarization in rabbit anterior mesenteric veins examined with a sucrose gap apparatus. Diazoxide also inhibited spontaneous electrical and contractile activity in guinea-pig taenia coli and in estrogen dominated rabbit uterus. In all these tissues, calcium is believed to play an important role in spontaneous electrical membrane activity. Diazoxide failed to affect contractility, rate of spontaneous contractions, or action potential configurations in isolated rabbit heart, even though the action potential in heart tissues possesses a definite calcium current component. Diazoxide reduced contractions induced in the mesenteric vein by electrical stimulation of the smooth muscle itself or by excitation of the nerve endings within the vein. Various drugs were chosen for their ability to contract the mesenteric vein in different ways. Noradrenaline contracts vascular smooth muscle even when the tissue Is depolarized with ouabain Diazoxide failed to inhibit noradrenaline contractions in the depolarized vein, but showed the characteristics of a competitive inhibitor of noradrenaline in normally polarized veins. Diazoxide was also capable of inhibiting contractions to serotonin and procaine, agents which require membrane polarization to initiate contraction. The inhibitory effect of diazoxide was not observed to be modified in solutions containing high concentrations of calcium. Diazoxide was tested upon the contractile responses to calcium In veins depolarized in K⁺ Ringer solution. Examination of the resultant dose response curves showed that diazoxide inhibited calcium contractions ln a reversible, non surmountable manner. Hydrochlorothiazide had no effect upon calcium induced contractions. Diazoxide antagonizes drug induced contractions only if a polarized membrane is present. Calcium Induced contractions in depolarizing solutions were inhibited in an apparently Insurmountable manner, while drug responses in polarizing solutions were inhibited by diazoxide in a surmountable manner. In addition, action potentials from rabbit heart were unchanged whereas, the apparently calcium spike mediated electrical activity of certain smooth muscles is inhibited. It is concluded that diazoxide affects the membrane of vascular smooth muscle to reduce excitability of the tissue to drugs or electrical stimuli. It is possible that cell membrane bound calcium could be the locus of action of diazoxide and that this agent modifies membrane calcium to cause increased membrane stability. / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate