A research report submitted to the Faculty of Health Sciences, University of Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Master of Science (MSc) in Pharmaceutical Affairs. / Lack of information has been identified as a major factor as to why patients do not take their medicines as the prescriber intends. Provision of appropriate information in a suitable form is therefore crucial. The package insert (PI) is the document that ensures the safe and effective use of the medicine under most circumstances. It presents a scientific, objective account of the medicine’s use as established by pre-clinical, clinical and often post-marketing studies. The patient information leaflet (PIL), which contains information for the consumer should be less scientific. South African legislation states that information contained in PILs must be aligned to PIs but the text must be readily intelligible for the patient. The study included a detailed comparison of prescribing information contained in the PI compared to the PIL in selected Angiotensin Receptor Blockers (ARBs). Findings of this comparative analysis revealed that key safety information was omitted from the PILs. An evaluation of the readability of the PILs was also performed by the use of Fry’s readability formula as well as applying elements of critical discourse analysis to determine if the texts in the PILs are suitable for its purpose. The results of the Fry’s readability assessments of all the PILs indicated that they had exceeded the recommended grade 7 reading level, which is in line with the adult literacy rate that qualifies anyone older than 15 years with a grade 7 qualification as being literate. Findings from the critical discourse analysis of the PILs show frequent use of medical jargon, complex sentence construction as well as ambiguity and slippage in the meaning of the texts in the PILs. The texts are not patient-friendly. Overall, the findings from this study indicate an urgent need to address the poor construction of PILs, to ensure that patients receive appropriate written prescribing information. This will ultimately ensure the safe and effective use of the medicine. / KP2020

Anita K.L. Yiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 93-112). / Acknowledgement --- p.i / List of Abbreviations --- p.ii / Abstract --- p.iv / Table of Contents --- p.vi / Chapter CHAPTER 1. --- Introduction / Chapter 1.1 --- The Renin-Angiotensin System (RAS) --- p.1 / Chapter 1.2 --- Physiology and Pathophysiology of Angiotensin --- p.3 / Chapter 1.3 --- Angiotensin Receptors / Chapter 1.3.1 --- Heterogeneity among Angiotensin Receptors --- p.10 / Chapter 1.3.2 --- Differential Distribution of Subtypes --- p.13 / Chapter 1.3.3 --- Molecular Structure of Subtypes --- p.15 / Chapter 1.3.4 --- Signal Transduction Mechanism --- p.20 / Chapter 1.3.5 --- Physiological Functional Correlates --- p.21 / Chapter 1.4 --- Aim of Study --- p.23 / Chapter CHAPTER 2. --- Preparation of Polyclonal Antibodies Against Angiotensin Receptors / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Preparation of antisera / Chapter 2.2.1.1 --- Preparation of peptide conjugates --- p.25 / Chapter 2.2.1.2 --- Protein determination --- p.27 / Chapter 2.2.1.3 --- Immunization of rabbits with peptide conjugates --- p.27 / Chapter 2.2.1.4 --- Collection of rabbit sera --- p.28 / Chapter 2.2.1.5 --- Affinity purification of antisera --- p.28 / Chapter 2.2.2 --- Enzyme-linked immunosorbent assay (ELISA) / Chapter 2.2.2.1 --- Titer determination --- p.29 / Chapter 2.2.2.2 --- Specificity determination --- p.30 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Preparation of antisera --- p.30 / Chapter 2.3.2 --- Affinity purification of antisera --- p.30 / Chapter 2.3.3 --- ELISA / Chapter 2.3.3.1 --- Titer determination --- p.31 / Chapter 2.3.3.1.1 --- Thy-AT1 antiserum --- p.31 / Chapter 2.3.3.1.2 --- Thy-AT2 antiserum --- p.32 / Chapter 2.3.3.2 --- Specificity determination --- p.32 / Chapter 2.3.3.2.1 --- Thy-AT1 antibodies --- p.32 / Chapter 2.3.3.2.2 --- Thy-AT2 antibodies --- p.49 / Chapter 2.4 --- Discussions --- p.49 / Chapter CHAPTER 3. --- Application of Thy-AT1 Antiserumin Western Blot / Chapter 3.1 --- Introduction --- p.52 / Chapter 3.2 --- Methods / Chapter 3.2.1 --- Preparation of protein samples --- p.52 / Chapter 3.2.2 --- Protein determination --- p.53 / Chapter 3.2.3 --- SDS-PAGE --- p.53 / Chapter 3.2.3 --- Western blot --- p.54 / Chapter 3.2.5 --- Immunoblotting --- p.54 / Chapter 3.3 --- Results --- p.55 / Chapter 3.4 --- Discussions --- p.58 / Chapter CHAPTER 4. --- Evaluation of Pancreatic Response to Angiotensin II / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.2 --- Methods / Chapter 4.2.1 --- Perfusion of pancreas --- p.62 / Chapter 4.2.2 --- Assay of amylase activity --- p.64 / Chapter 4.2.3. --- Calculations --- p.64 / Chapter 4.3 --- Results --- p.65 / Chapter 4.4 --- Discussions --- p.65 / Chapter CHAPTER 5. --- Application of Purified Thy-AT2 Antibodies in immunohistochemical studies / Chapter 5.1 --- Introduction --- p.74 / Chapter 5.2 --- Methods / Chapter 5.2.1 --- Preparation of adrenal sections --- p.75 / Chapter 5.2.2 --- Light-microscopic immunohistochemical study --- p.76 / Chapter 5.3 --- Results / Chapter 5.4 --- Discussions / Chapter CHAPTER 6. --- General Discussions --- p.84 / References --- p.93 / Appendix / Chapter A. --- Materials --- p.113 / Chapter B. --- Buffer Compositions --- p.121

by Chi-shing Sum. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 84-96). / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / LIST OF ABBREVIATIONS --- p.iv / TABLE OF CONTENTS --- p.v / Chapter CHAPTER 1 --- p.1 / Chapter 1.1 --- Biochemistry of Renin-Angiotensin System --- p.1 / Chapter 1.2 --- Physiological Roles of Angiotensin --- p.5 / Chapter 1.3 --- Biochemistry of Angiotensin Receptors --- p.6 / Chapter 1.4 --- Tissue Renin-Angiotensin System --- p.13 / Kidney --- p.13 / Blood vessels --- p.14 / Heart --- p.15 / Brain --- p.16 / Testes --- p.17 / Chapter 1.5 --- Structure and Function of Vas Deferens --- p.18 / Chapter 1.6 --- Aim of Study --- p.21 / Chapter CHAPTER 2 --- p.22 / Chapter 2.1 --- introduction --- p.22 / Chapter 2.2 --- Materials --- p.23 / Chapter 2.3 --- methods --- p.23 / Chapter 2.3.1 --- Preparation of isolated epididymal rat vas deferens --- p.23 / Chapter 2.3.2 --- Concentration-responses to angiotensins --- p.24 / Chapter 2.3.3 --- Effects of angiotensin II in the presence of protease inhibitors --- p.24 / Chapter 2.3.4 --- Effect of losartan and CGP 42112 --- p.24 / Chapter 2.3.5 --- Schild analysis --- p.25 / Chapter 2.3.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.25 / Chapter 2.3.7 --- Statistical analysis --- p.25 / Chapter 2.4 --- results --- p.25 / Chapter 2.4.1 --- Effect of angiotensin on epididymal rat vas deferens --- p.25 / Chapter 2.4.2 --- Concentration-responses to angiotensins in epididymal rat vas deferens --- p.27 / Chapter 2.4.3 --- Effect of angiotensin II in the presence of protease inhibitors --- p.27 / Chapter 2.4.4 --- Effect of losartan ami CGP 42112 --- p.27 / Chapter 2.4.5 --- Schild analysis --- p.36 / Chapter 2.4.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.36 / Chapter 2.5 --- Discussion --- p.36 / Chapter CHAPTER 3 --- p.39 / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Materials --- p.39 / Chapter 3.3 --- Methods --- p.40 / Chapter 3.3.1 --- Preparation of isolated prostatic rat vas deferens --- p.40 / Chapter 3.3.2 --- Concentration-responses to angiotensins --- p.40 / Chapter 3.3.3 --- Effects of angiotensin II in the presence of protease inhibitors --- p.41 / Chapter 3.3.4 --- Effect oflosartan and CGP 42112 --- p.41 / Chapter 3.3.5 --- Schild analysis --- p.41 / Chapter 3.3.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.42 / Chapter 3.3.7 --- Concentration-response to angiotensin II after reserpine treatment --- p.42 / Chapter 3.3.8 --- Concentration-response to angiotensin II after desensitization of P2-purinoceptors --- p.42 / Chapter 3.3.9 --- Statistical analysis --- p.42 / Chapter 3.4 --- Results --- p.43 / Chapter 3.4.1 --- Effect of angiotensin on prostatic rat vas deferens --- p.43 / Chapter 3.4.2 --- Concentration-responses to angiotensins in prostatic rat vas deferens --- p.43 / Chapter 3.4.3 --- Effect of angiotensin II in the presence of protease inhibitors --- p.43 / Chapter 3.4.4 --- Effect of losartan and CGP 42112 --- p.49 / Chapter 3.4.5 --- Schild analysis --- p.49 / Chapter 3.4.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.49 / Chapter 3.4.7 --- Concentration-response to angiotensin II afier reserpine treatment --- p.54 / Chapter 3.4.8 --- Concentration-response to angiotensin II after desensitization of P2-purinoceptors --- p.54 / Chapter 3.5 --- Discussion --- p.63 / Chapter CHAPTER 4 --- p.66 / Chapter 4.1 --- introduction --- p.66 / Chapter 4.2 --- Materials and Methods --- p.66 / Chapter 4.2.1 --- Preparation of polyclonal angiotensin II antiserum --- p.66 / Chapter 4.2.1.1 --- Preparation of peptide conjugate --- p.66 / Chapter 4.2.1.2 --- Protein determination --- p.67 / Chapter 4.2.1.3 --- Immunization of rabbit with peptide conjugate --- p.67 / Chapter 4.2.1.4 --- Collecting rabbit serum --- p.68 / Chapter 4.2.2 --- Characterization of BSA-Ang II and Thy-Ang II antisera --- p.68 / Chapter 4.2.2.1 --- Slot blotting --- p.68 / Chapter 4.2.2.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.69 / Chapter 4.3 --- RESULT --- p.69 / Chapter 4.3.1 --- Preparation of polyclonal angiotensin II antiserum --- p.69 / Chapter 4.3.2 --- Characterization of BSA-Ang II and Thy-Ang II antisera --- p.70 / Chapter 4.3.2.1 --- Slot blotting --- p.70 / Chapter 4.3.2.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.70 / Chapter 4.3 --- discussion --- p.76 / Chapter CHAPTER 5 --- p.78 / Chapter 5. 1 --- General Discussions --- p.78 / REFERENCES --- p.84 / APPENDIX --- p.97 / Published Abstract and Paper --- p.97

Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.

Tang Wai-man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 142-147). / Abstract also in Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgement --- p.v / Lists of Abbreviations --- p.vi / Table of Contents --- p.vii / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Isolation of Mas Oncogene --- p.1 / Chapter 1.2 --- Distribution of Mas Oncogene..........…… --- p.3 / Chapter 1.3 --- Developmental Expression of Mas Oncogene --- p.5 / Chapter 1.4 --- Study of Mas-deficient Mice --- p.7 / Chapter 1.5 --- Signal Transduction of Mas Oncogene --- p.8 / Chapter 1.6 --- Other Family Member of Mas Oncogene --- p.9 / Chapter 1.7 --- Mas and Angiotensin Receptor --- p.11 / Chapter 1.8 --- Angiotensin Receptors / Chapter 1.8.1 --- Classification of Angiotensin AT1 Receptor --- p.14 / Chapter 1.8.2 --- Cloning of Angiotensin Receptor --- p.15 / Chapter 1.9 --- Expression of Angiotensin Receptor / Chapter 1.9.1 --- Physiological Factors --- p.17 / Chapter 1.9.2 --- Cis-regulatory Elements / Chapter 1.9.2.1 --- Organization and Regulatory Elements of AT1 Receptor --- p.19 / Chapter 1.9.2.2 --- Expression of AT1a Receptor Promoter was Induced by AP-1 and GATA-4 in Pressure Overload Model --- p.20 / Chapter 1.9.2.3 --- AT1a Receptor Reveals Three Glucocorticoid Responsive Elements --- p.22 / Chapter 1.10 --- Signal Transduction of Angiotensin Receptor --- p.22 / Chapter 1.11 --- Aim of Project --- p.25 / Chapter Chapter 2: --- Mas Oncogene in AR4-2J cells / Chapter 2.1 --- Introduction --- p.26 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Materials / Chapter 2.2.1.1 --- Reagents --- p.27 / Chapter 2.2.1.2 --- Enzymes --- p.27 / Chapter 2.2.1.3 --- DNA Purification Kits --- p.28 / Chapter 2.2.1.4 --- Materials and Antibodies for Western Blot --- p.28 / Chapter 2.2.1.5 --- Others --- p.28 / Chapter 2.2.2 --- Restriction Enzyme Digestion --- p.29 / Chapter 2.2.3 --- Agarose Gel Electrophoresis --- p.29 / Chapter 2.2.4 --- DNA Extraction and Purification --- p.29 / Chapter 2.2.5 --- Plasmid Vector Modification and DNA Ligation --- p.30 / Chapter 2.2.6 --- Bacterial Transformation --- p.31 / Chapter 2.2.7 --- Preparation of Plasmid DNA / Chapter 2.2.7.1 --- Minipreps --- p.32 / Chapter 2.2.7.2 --- Midipreps and Maxipreps --- p.33 / Chapter 2.2.8 --- Genomic DNA Extraction From Tissue and Cell Culture --- p.34 / Chapter 2.2.9 --- RT-PCR Cloning of Mas Oncogene --- p.35 / Chapter 2.2.10 --- Construction of Full Length Mas cDNA into pBluescript® II SK Vector --- p.38 / Chapter 2.2.11 --- Southern Blot Analysis / Chapter 2.2.11.1 --- Preparation of DIG-labeled Mas Probe --- p.38 / Chapter 2.2.11.2 --- Enzyme Restriction of Genomic DNA --- p.39 / Chapter 2.2.11.3 --- Transferring DNA to Nylon Membrane --- p.40 / Chapter 2.2.11.4 --- Prehybridization and Hybridization --- p.40 / Chapter 2.2.11.5 --- Post-hybridization Washes and Blocking --- p.41 / Chapter 2.2.11.6 --- Detection --- p.41 / Chapter 2.2.12 --- DNA Sequencing / Chapter 2.2.12.1 --- Manual Sequencing --- p.42 / Chapter 2.2.12.2 --- Autosequencing --- p.43 / Chapter 2.2.12.3 --- Sequencing Primers --- p.44 / Chapter 2.2.13 --- Cell Culture --- p.45 / Chapter 2.2.14 --- Protein Assay by Modified Lowery --- p.46 / Chapter 2.2.15 --- SDS-PAGE and Western Blot Analysis --- p.47 / Chapter 2.3 --- Results --- p.49 / Chapter 2.4 --- Discussion --- p.60 / Chapter Chapter 3: --- Analysis of Transfected Mas Cell Lines / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Materials --- p.62 / Chapter 3.2.2 --- Cell Culture and Transfection / Chapter 3.2.2.1 --- Cell Culture --- p.62 / Chapter 3.2.2.2 --- Transfection Optimization --- p.62 / Chapter 3.2.2.3 --- Fluorescent SEAP Assay --- p.63 / Chapter 3.2.2.4 --- Transient Transfection --- p.64 / Chapter 3.2.2.5 --- Stable Cell Line Construction --- p.64 / Chapter 3.2.3 --- Protein Assay ESL --- p.65 / Chapter 3.2.4 --- SDS-PAGE and Western Blot Analysis --- p.65 / Chapter 3.2.5 --- Preparation of an AT1a Receptor Internal Standard for Quantitative RT-PCR Analysis / Chapter 3.2.5.1 --- Preparation of an AT1a Receptor cDNA by RT-PC --- p.66 / Chapter 3.2.5.2 --- Cloning of AT1A Receptor cDNA into pBluescript® II SK Vector --- p.67 / Chapter 3.2.5.3 --- Autosequence of pBluescript® II SK Vector/AT1AR --- p.68 / Chapter 3.2.5.4 --- Preparation of 100 bp Deleted AT1a Receptor cDNA by RT- PCR --- p.68 / Chapter 3.2.5.5 --- Cloning of Deleted AT1a R cDNA into pCAPs Vector --- p.71 / Chapter 3.2.6 --- Construction of Full Length Mas cDNA into pOPRSVI/MCS Operator Vector --- p.71 / Chapter 3.2.7 --- Preparation of an Mas Internal Standard for Quantitative RT-PCR Analysis / Chapter 3.2.7.1 --- Preparation of 100 bp Deleted Mas cDNA by RT- PCR --- p.72 / Chapter 3.2.7.2 --- Cloning of 100 bp Deleted Mas cDNA into pCAPs Vector (Mas/pCAPs) --- p.73 / Chapter 3.2.8 --- Quantitative RT-PCR Analysis of AT1A R Expression --- p.74 / Chapter 3.2.9 --- Quantitative RT-PCR Analysis for the Expression of Mas --- p.74 / Chapter 3.3 --- Results --- p.76 / Chapter 3.4 --- Discussions --- p.100 / Chapter Chapter 4: --- Cloning of AT1A Receptor Promoter / Chapter 4.1 --- Introduction --- p.104 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Materials --- p.105 / Chapter 4.2.2 --- Genomic DNA Extraction From Rat Pancreas --- p.105 / Chapter 4.2.3 --- "Nest PCR Amplification of 3.2, 2.8 and 1.4kb AT1a Receptor Promoter" --- p.105 / Chapter 4.2.4 --- PCR Amplification of 2.2 kb Aproximal Portion of AT1a Receptor Promoter --- p.107 / Chapter 4.2.5 --- Construction of PCR Fragment of Angiotensin Receptor Promoter into Various Vector --- p.108 / Chapter 4.2.5.1 --- pSEAP2-Basic --- p.108 / Chapter 4.2.5.2 --- pBluescript® II SK Vector --- p.109 / Chapter 4.2.5.3 --- PCR Cloning Kit (pCAPs vector) --- p.109 / Chapter 4.2.5.4 --- PCR-TRAP Cloning System --- p.109 / Chapter 4.2.6 --- Direct PCR Analysis --- p.110 / Chapter 4.2.7 --- Autosequencing of PCR Fragment of AT1A Receptor Promoter --- p.111 / Chapter 4.3 --- Results --- p.114 / Chapter 4.4 --- Discussions --- p.130 / Chapter Chapter 5: --- General Discussion --- p.131 / Chapter Appendix 1 --- Composition of Solutions --- p.133 / Chapter Appendix 2 --- Published Abstract --- p.141 / References --- p.142

Siu-Yin Sylvia Lam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 123-140). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iv / 英中譯名對照 --- p.vi / Acknowledgements --- p.vii / Table of Contents --- p.viii / Abbreviations --- p.xiii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Overview of Carotid Body --- p.1 / Chapter 1.1.1 --- Type I Cells --- p.3 / Chapter 1.1.2 --- Type II Cells --- p.4 / Chapter 1.1.3 --- Blood Vessels --- p.5 / Chapter 1.1.4 --- Innervation --- p.5 / Chapter 1.1.5 --- Biochemistry --- p.6 / Chapter 1.1.6 --- Physiology and Function --- p.7 / Chapter 1.2 --- The Renin-Angiotensin System (RAS) --- p.8 / Chapter 1.2.1 --- Circulating RAS --- p.8 / Chapter 1.2.1.1 --- Angiotensinogen --- p.10 / Chapter 1.2.1.2 --- Renin --- p.10 / Chapter 1.2.1.3 --- Angiotensin I --- p.11 / Chapter 1.2.1.4 --- Angiotensin Converting Enzyme --- p.12 / Chapter 1.2.1.5 --- Angiotensin II --- p.12 / Chapter 1.2.1.6 --- Angiotensin II Receptors --- p.13 / Chapter 1.2.1.7 --- Angiotensin IV and Angiotensin IV Receptor --- p.15 / Chapter 1.2.2 --- Tissue RAS --- p.16 / Chapter 1.3 --- Hypoxia and Carotid Body --- p.18 / Chapter 1.4 --- Hypoxia and RAS --- p.21 / Chapter 1.5 --- Hypoxia and RAS in Carotid Body --- p.23 / Chapter 1.6 --- Aims of Study --- p.24 / Chapter 1.6.1 --- Existence of Functional Angiotensin II Receptors --- p.24 / Chapter 1.6.2 --- Regulation and Function of Angiotensin II Receptors by Chronic Hypoxia --- p.24 / Chapter 1.6.3 --- Existence of an Intrinsic Angiotensin-generating System --- p.25 / Chapter 1.6.4 --- Regulation of Local RAS by Chronic Hypoxia --- p.25 / Chapter 1.6.5 --- Studies of AT4 Receptor --- p.26 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Experimental Animals and Rat Models --- p.27 / Chapter 2.1.1 --- Rat Model of Chronic Hypoxia --- p.27 / Chapter 2.1.2 --- Isolation of Carotid Body --- p.28 / Chapter 2.2 --- Semi-quantitative Reverse Transcriptase-polymerase Chain Reaction (RT-PCR) --- p.30 / Chapter 2.2.1 --- Total RNA Extraction and Quantification --- p.30 / Chapter 2.2.2 --- Reverse Transcription (RT) --- p.31 / Chapter 2.2.3 --- Polymerase Chain Reaction (PCR) --- p.31 / Chapter 2.2.4 --- Gel Electrophoresis --- p.34 / Chapter 2.2.5 --- Optimization of Semi-quantitative RT-PCR for RAS Gene Analysis --- p.34 / Chapter 2.3 --- Northern Blotting --- p.35 / Chapter 2.3.1 --- Transfer of Denatured RNA to Nitrocellulose Membrane By Capillary Elution --- p.35 / Chapter 2.3.2 --- Hybridization --- p.36 / Chapter 2.4 --- In-situ Hybridization --- p.38 / Chapter 2.4.1 --- Linearization of Angiotensinogen cDNA --- p.38 / Chapter 2.4.2 --- Riboprobe Preparation --- p.38 / Chapter 2.4.3 --- Quantification and Gel Electrophoresis of Riboprobes --- p.39 / Chapter 2.4.4 --- In-situ Hybridization Histochemistry --- p.39 / Chapter 2.5 --- Immunohistochemistry --- p.42 / Chapter 2.5.1 --- Preparation of Cryosection --- p.42 / Chapter 2.5.2 --- Indirect Immunoperoxidase Staining --- p.42 / Chapter 2.5.3 --- Immunofluorescent Double Staining --- p.43 / Chapter 2.6 --- Western Blot Analysis --- p.45 / Chapter 2.6.1 --- Preparation of Angiotensinogen Protein --- p.45 / Chapter 2.6.2 --- Quantification of Protein Concentration --- p.45 / Chapter 2.6.3 --- Sample Preparation --- p.45 / Chapter 2.6.4 --- Sodium Dodecyl-sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.46 / Chapter 2.6.5 --- Electroblotting and Immunodetection of Proteins --- p.46 / Chapter 2.7 --- Spectrofluorimetric Measurement and In-vitro Electrophysiology --- p.48 / Chapter 2.7.1 --- Dissociation of Carotid Body Type I Cells and Spectrofluorimetric Measurement --- p.48 / Chapter 2.7.2 --- In-vitro Electrophysiology --- p.49 / Chapter 2.8 --- Assay of ACE Activity --- p.51 / Chapter 2.8.1 --- Crude Membrane Preparation --- p.51 / Chapter 2.8.2 --- Basic Principle for ACE Activity Measurement --- p.51 / Chapter 2.8.3 --- Measurement of ACE Activity --- p.51 / Chapter 2.8.4 --- Fluorescence Measurement --- p.53 / Chapter 2.9 --- In-vitro Autoradiography and Fluorescence-labeled Binding Assay for Angiotensin IV --- p.54 / Chapter 2.9.1 --- Preparation of Frozen Tissue Sections --- p.54 / Chapter 2.9.2 --- Localization and Density of AT4 Receptor --- p.54 / Chapter 2.10 --- Statistics and Data Analysis --- p.57 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Functional Expression of Angiotensin II Receptors --- p.58 / Chapter 3.1.1 --- [Ca2+]i Response to Angiotensin II --- p.58 / Chapter 3.1.2 --- Antagonistic Blockade of Angiotensin II Receptor Subtypes --- p.58 / Chapter 3.1.3 --- Expression of AT1 Receptors mRNA --- p.61 / Chapter 3.1.4 --- Cellular Localization of AT1 Receptors Protein --- p.61 / Chapter 3.2 --- Effect of Chronic Hypoxia on the Expression and Function of Angiotensin II Receptors --- p.64 / Chapter 3.2.1 --- Effect of Chronic Hypoxia on the Expression of AT1 Receptors --- p.64 / Chapter 3.2.2 --- Effect of Chronic Hypoxia on the Expression of AT2 Receptors --- p.67 / Chapter 3.2.3 --- Cellular Localization of the AT1 Receptor by Chronic Hypoxia --- p.69 / Chapter 3.2.4 --- Increase of Afferent Nerve Activities of the Carotid Body In-vitro by Angiotensin II --- p.71 / Chapter 3.2.5 --- Inhibition of Angiotensin II-mediated Response in Chronically Hypoxic Carotid Body by Losartan --- p.73 / Chapter 3.3 --- Evidence for the Existence of an Intrinsic Angiotensin-generating System --- p.75 / Chapter 3.3.1 --- Expression and Localization of Angiotensinogen mRNA --- p.75 / Chapter 3.3.2 --- Expression and Localization of Angiotensinogen Protein --- p.78 / Chapter 3.3.3 --- Expression of Renin mRNA --- p.81 / Chapter 3.3.4 --- Expression of ACE mRNA --- p.81 / Chapter 3.4 --- Effect of Chronic Hypoxia on the Locally-generated Angiotensin System --- p.85 / Chapter 3.4.1 --- Effect of Chronic Hypoxia on the Expression of Angiotensinogen mRNA --- p.85 / Chapter 3.4.2 --- Effect of Chronic Hypoxia on the Localization of Angiotensinogen mRNA --- p.87 / Chapter 3.4.3 --- Effect of Chronic Hypoxia on the Expression of Angiotensinogen Protein --- p.89 / Chapter 3.4.4 --- Effect of Chronic Hypoxia on the Expression of ACE --- p.91 / Chapter 3.5 --- Time-course Effect of Chronic Hypoxia on ACE Activity --- p.93 / Chapter 3.6 --- Preliminary Studies of AT4 Receptor --- p.98 / Chapter 3.6.1 --- In-vitro Autoradiographic Study of AT4 Receptors --- p.98 / Chapter 3.6.2 --- Localization of AT4 Receptors --- p.100 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Functional Expression of Angiotensin II Receptors --- p.102 / Chapter 4.2 --- Upregulation and Function of Angiotensin II Receptors --- p.105 / Chapter 4.3 --- Existence of a Local RAS --- p.108 / Chapter 4.4 --- Regulation of the Local RAS --- p.112 / Chapter 4.5 --- Time-dependent Changes of ACE Activity --- p.155 / Chapter 4.6 --- Presence and Regulation of AT4 Receptor --- p.117 / Chapter 4.7 --- Conclusion --- p.120 / Chapter 4.8 --- Future Works --- p.121 / Chapter Chapter 5 --- References --- p.123

by Pui-fan Wong. / "December 2001." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / 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.

Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.

Made available in DSpace on 2014-06-11T19:23:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-27Bitstream added on 2014-06-13T18:51:03Z : No. of bitstreams: 1 gealh_wc_me_araca.pdf: 769445 bytes, checksum: 471ed1e28e30d4bd94713ea173fc064a (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A hipertensão arterial é o maior fator de risco para doenças cardiovasculares, caracterizada por uma disfunção endotelial, alteração no controle do crescimento das células vasculares e morte celular. Alterações como gradual perda óssea e diminuição no índice de massa óssea corpórea têm sido fortemente relacionadas com hipertensão, sendo que os mecanismos celulares e moleculares que envolvem a hipertensão e osteoporose não são compreendidos completamente como um todo. Objetivos: O presente projeto teve por objetivo estudar a interferência da hipertensão essencial no metabolismo ósseo durante o processo de reparo de enxerto ósseo autógeno, bem como analisar a influência do losartan, um medicamento anti-hipertensivo antagonista dos receptores de angiotensina, analisando a expressão das proteínas OC, OPG, RANK, RANKL, TRAP, VEGF e PECAM através da técnica de imunoistoquímica. Além disso, este projeto pretendeu realizar a análise histomorfométrica do tecido ósseo formado durante o processo de reparo. Material e métodos: Os grupos estudados compreenderam 24 ratos normotensos Wistar e 24 ratos espontaneamente hipertensos, divididos em grupos não tratados e tratados com Losartan, submetidos a procedimentos cirúrgicos para realização de enxertos ósseos em bloco na mandíbula, nos períodos de 7, 14 e 28 dias. Conclusão: Conclui-se que os animais SHR apresentam atraso no processo de reparo de enxertos ósseos autógeno em bloco quando comparados aos animais Wistar, e que o uso do losartan para redução da pressão arterial nestes animais demonstrou melhorar o processo cicatricial, apesar de não haverem diferenças estatísticas significantes / Hypertension is the major risk factor to coronary diseases characterized by endothelial disfunction, alterations on the growing of vascular cell and death cell. Alterations like continuous bone loss and lower body mass index have been strongly connected with hypertension, althought the mollecular and cellular mechanisms wich involve the hypertension and osteoporosis are not completely understood. Objectives: This study has the objective to observe the high blood pressure interference in the osseous metabolism during the bone healing of autogenous bone graft and carry out analysis about the administration of losartan, a receptor angiotensin antagonist by OC, OPG, RANK, RANKL, TRAP, VEGF and PECAM expressions by immunohistochemical technique and hystological analysis of the bone tissue. Material and methods: The groups studied were 24 normotensive rats Wistar and 24 spontaneous hypertensive rats dealed out non treated and treated groups by losartan. Surgical procedures were carried out and an onlay bone grat were fixed on the mandible and the animmals were sacrified on the 7, 14 and 28 days. Conclusion: we conclued that the SHR has a late bone healing process when comparing with Wistar group and the losartan administration to reduce the blood pressure has the potential to become better the bone graft healing, althought significant estatistical differences not exist

Orientador: Roberta Okamoto / Coorientador: Cristina Antoniali Silva / Banca: Idelmo Rangel Garcia Júnior / Banca: Edevaldo Tadeu Camarini / Resumo: A hipertensão arterial é o maior fator de risco para doenças cardiovasculares, caracterizada por uma disfunção endotelial, alteração no controle do crescimento das células vasculares e morte celular. Alterações como gradual perda óssea e diminuição no índice de massa óssea corpórea têm sido fortemente relacionadas com hipertensão, sendo que os mecanismos celulares e moleculares que envolvem a hipertensão e osteoporose não são compreendidos completamente como um todo. Objetivos: O presente projeto teve por objetivo estudar a interferência da hipertensão essencial no metabolismo ósseo durante o processo de reparo de enxerto ósseo autógeno, bem como analisar a influência do losartan, um medicamento anti-hipertensivo antagonista dos receptores de angiotensina, analisando a expressão das proteínas OC, OPG, RANK, RANKL, TRAP, VEGF e PECAM através da técnica de imunoistoquímica. Além disso, este projeto pretendeu realizar a análise histomorfométrica do tecido ósseo formado durante o processo de reparo. Material e métodos: Os grupos estudados compreenderam 24 ratos normotensos Wistar e 24 ratos espontaneamente hipertensos, divididos em grupos não tratados e tratados com Losartan, submetidos a procedimentos cirúrgicos para realização de enxertos ósseos em bloco na mandíbula, nos períodos de 7, 14 e 28 dias. Conclusão: Conclui-se que os animais SHR apresentam atraso no processo de reparo de enxertos ósseos autógeno em bloco quando comparados aos animais Wistar, e que o uso do losartan para redução da pressão arterial nestes animais demonstrou melhorar o processo cicatricial, apesar de não haverem diferenças estatísticas significantes / Abstract: Hypertension is the major risk factor to coronary diseases characterized by endothelial disfunction, alterations on the growing of vascular cell and death cell. Alterations like continuous bone loss and lower body mass index have been strongly connected with hypertension, althought the mollecular and cellular mechanisms wich involve the hypertension and osteoporosis are not completely understood. Objectives: This study has the objective to observe the high blood pressure interference in the osseous metabolism during the bone healing of autogenous bone graft and carry out analysis about the administration of losartan, a receptor angiotensin antagonist by OC, OPG, RANK, RANKL, TRAP, VEGF and PECAM expressions by immunohistochemical technique and hystological analysis of the bone tissue. Material and methods: The groups studied were 24 normotensive rats Wistar and 24 spontaneous hypertensive rats dealed out non treated and treated groups by losartan. Surgical procedures were carried out and an onlay bone grat were fixed on the mandible and the animmals were sacrified on the 7, 14 and 28 days. Conclusion: we conclued that the SHR has a late bone healing process when comparing with Wistar group and the losartan administration to reduce the blood pressure has the potential to become better the bone graft healing, althought significant estatistical differences not exist / Mestre