by Mano Arumanayagam. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 249-265. / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Transport Pathways of Sodium in Human Erythrocytes --- p.2 / Chapter 1.2 --- Active Transport - the Sodium Pump --- p.2 / Chapter 1.2.1 --- Effects of Sodium and Potassium Ions --- p.3 / Chapter 1.2.2 --- Molecular Weight and Subunit Structure --- p.4 / Chapter 1.2.3 --- Inhibitors of the Sodium Pump --- p.6 / Chapter 1.2.4 --- "Measurements of the Sodium Pump/Na+,K+-ATPase" --- p.8 / Chapter 1.3 --- The Passive Fluxes of Sodium & Potassium --- p.13 / Chapter 1.3.1 --- The Na-K-Cl Co-transport System (SPC) --- p.14 / Chapter 1.3.2 --- Sodium-Lithium Countertransport (SLC) --- p.16 / Chapter 1.3.3 --- Ouabain & Frusemide Insensitive Na+ 'leak' --- p.17 / Chapter 1.3.4 --- Methods for the Determination of Passive Na Fluxes --- p.18 / Chapter 1.4 --- Erythrocyte Sodium Transport and Thyroid Hormones --- p.19 / Chapter 1.5 --- Thyroid Hormones and the Na Pump in Other Cells --- p.22 / Chapter 1.6 --- "Mechanism of T3 Induced Increments of the Sodium Pump/Na+,K+-ATPase Activity" --- p.25 / Chapter 1.7 --- Aims of the Project --- p.28 / Chapter CHAPTER 2 --- "ERYTHROCYTE SODIUM FLUXES, OBS AND NA+,K+-ATPASE ACTIVITY IN HYPERTHYROIDISM 29" / Chapter 2.1 --- PREAMBLE --- p.30 / Chapter 2.2 --- MATERIALS & METHODS --- p.31 / Chapter 2.2.1 --- Materials --- p.31 / Chapter 2.2.2 --- Subjects --- p.31 / Chapter 2.2.3 --- Blood Specimen --- p.32 / Chapter 2.2.4 --- Separation of Erythrocytes from Whole Blood --- p.32 / Chapter 2.2.5 --- Erythrocyte Naic and Kic --- p.32 / Chapter 2.2.6 --- Ouabain-sensitive sodium transport --- p.33 / Chapter 2.2.7 --- "Determination of Na+,K+-ATPase activity" --- p.34 / Chapter 2.2.8 --- "Determination SPC, SLC and Na+ 'leak'" --- p.35 / Chapter 2.2.9 --- Determination of OBS and its Kd --- p.37 / Chapter 2.2.10 --- Plasma Thyroid Hormone Analyses --- p.41 / Chapter 2.2.11 --- "Determination of Plasma Concentrations of Sodium, Potassium, Urea and Creatinine" --- p.41 / Chapter 2.2.12 --- "Determination of Hb concentration, Leucocyte and Erythrocyte Counts, and MCV" --- p.41 / Chapter 2.2.13 --- Assessment of the Precision of the Methods --- p.41 / Chapter 2.3 --- STATISTICS --- p.42 / Chapter 2.4 --- RESULTS --- p.44 / Chapter 2.4.1 --- Determination of the Concentration of Ouabain in a Stock of 3H-ouabain --- p.44 / Chapter 2.4.1.1 --- Effect of Incubation Time on Binding of Ouabain to Erythrocytes --- p.46 / Chapter 2.4.1.2 --- Scatchard Plots of Bound Against Ratio of Bound to Free --- p.48 / Chapter 2.4.2 --- "Establishment of the Method for the Simultaneous Determination of SLC, SPC and Na+'leak'" --- p.48 / Chapter 2.4.3 --- Descriptive Statistics of the Subjects in the Study --- p.54 / Chapter 2.4.4 --- "Plasma concentrations of T4, T3, free T4, and free T3 in control subjects and hyperthyroid patients" --- p.54 / Chapter 2.4.5 --- "Plasma Concentrations of Sodium, Potassium, Urea, Creatinine and Ratios of Creatinine to Urea in Control Subjects and Hyperthyroid Patients" --- p.54 / Chapter 2.4.6 --- "Leucocyte Count, Erythrocyte Count, Hb Concentration, MCV and Derived Values for PCV, MCHC and MCH in Control Subjects and Hyperthyroid Patients" --- p.58 / Chapter 2.4.7 --- "Intracellular Sodium Naic, potassium Kic, ouabain-sensitive efflux rate (fo), ouabain-sensitive efflux rate constant (ko), OBS and its dissociation constant (Kd), and Na+,K+-ATPase activity in control subjects and hyperthyroid patients" --- p.58 / Chapter 2.4.8 --- SPC and its Rate Constant --- p.58 / Chapter 2.4.9 --- SLC and its rate constant --- p.63 / Chapter 2.4.10 --- Na+ 'leak' and its rate constant --- p.63 / Chapter 2.4.11.1 --- The Spearman Rank Coefficient of Correlation Matrix for the Characteristics of Sodium Transport Before Treatment --- p.63 / Chapter 2.4.11.2 --- Partial Coefficient of Correlation Between the Rate Constant for SLC and OBS and Naic --- p.69 / Chapter 2.4.11.3 --- Partial Coefficients of Correlation Between the Rate Constants for SLC and SPC with Naic Held Constant --- p.71 / Chapter 2.4.11.4 --- "Partial Coefficients of Correlation (r123) Between Na+ 'leak' and OBS, Na+,K+-ATPase activity and efflux rate constant with Naic Held Constant" --- p.71 / Chapter 2.4.12 --- Spearman Rank Coefficient of Correlation for Plasma Thyroid Function Tests with Sodium Transport Variables --- p.71 / Chapter 2.4.13 --- Effect of Treatment on 11 of the 18 Subjects --- p.74 / Chapter 2.4.13.1 --- "Effect of Treatment on Body Weight, Systolic and Diastolic BP's" --- p.74 / Chapter 2.4.13.2 --- "Plasma Concentrations of Sodium, Potassium, Urea, Creatinine and Ratio of Urea to Creatinine After Treatment" --- p.74 / Chapter 2.4.13.3 --- Hb Concentrations and Other Blood Indices Before and After Treatment --- p.74 / Chapter 2.4.13.4 --- Plasma Concentrations of T4 and Free T3 in patients before and after treatment --- p.78 / Chapter 2.4.13.5 --- The Characteristics of the Sodium Pump --- p.78 / Chapter 2.4.13.6 --- Passive Fluxes of Sodium After Treatment --- p.78 / Chapter 2.4.14 --- "Longitudinal Assessment of Plasma Thyroid Function Tests, Naic, Kic, OBS, Na+,K + -ATPase Activity and Sodium Fluxes in Patients Undergoing Treatment" --- p.87 / Chapter 2.5 --- DISCUSSION --- p.95 / Chapter CHAPTER 3 --- "THE EFFECT OF HYPERTHYROIDISM ON IN VIVO AGING OF ERYTHROCYTE OUABAIN BINDING SITES, INTRACELLULAR SODIUM AND POTASSIUM CONCENTRATIONS 105" / Chapter 3.1 --- Review of Literature --- p.106 / Chapter 3.1.1 --- Physical Changes and Methods of Separation --- p.106 / Chapter 3.1.2 --- Biochemical Changes --- p.108 / Chapter 3.1.2.1 --- "Naic,Kic and the Transport of Sodium" --- p.108 / Chapter 3.1.2.2 --- Changes in Other Enzymes/Proteins --- p.111 / Chapter 3.2 --- MATERIALS & METHODS --- p.116 / Chapter 3.2.1 --- Materials --- p.116 / Chapter 3.2.2 --- Subjects --- p.116 / Chapter 3.2.3 --- Plasma Thyroid Hormone Analyses --- p.116 / Chapter 3.2.4 --- Separation of Erythrocytes According to Age --- p.117 / Chapter 3.2.5 --- Determination of MCV & MCHC --- p.118 / Chapter 3.2.6 --- Erythrocyte Creatine Concentration --- p.118 / Chapter 3.2.7 --- Determination of Naic and Kic --- p.119 / Chapter 3.2.8 --- Maximum Number of OBS --- p.119 / Chapter 3.3 --- STATISTICS --- p.119 / Chapter 3.4 --- RESULTS --- p.120 / Chapter 3.4.1 --- "Establishment of the Method for the Separation of Young, Middle and Old Cells" --- p.120 / Chapter 3.4.2 --- Descriptive Statistics of the Subjects in the Study --- p.122 / Chapter 3.4.4 --- Effect of Erythrocyte Age on Markers of Cell Age --- p.124 / Chapter 3.4.4 --- "Effect of Erythrocyte Age on Naic, Kic and OBS" --- p.127 / Chapter 3.5 --- DISCUSSION --- p.133 / Chapter CHAPTER 4 --- THYROID HORMONES AND OUABAIN BINDING SITES OF RETICULOCYTES --- p.140 / Chapter 4.1.1 --- Review of the Literature --- p.141 / Chapter 4.1.2 --- Intracellular Organelles --- p.143 / Chapter 4.1.2.1 --- Ribosomes & RNA --- p.143 / Chapter 4.1.2.2 --- Mitochondria --- p.143 / Chapter 4.1.2.3 --- Other Organelles --- p.144 / Chapter 4.1.3 --- Changes in the Sodium Pump --- p.145 / Chapter 4.1.4 --- Changes in Other Membrane Proteins --- p.146 / Chapter 4.1.5 --- Aim of the Study --- p.147 / Chapter 4.2 --- METHODS --- p.148 / Chapter 4.2.1 --- Animals --- p.148 / Chapter 4.2.2 --- Induction of Reticulocytosis --- p.148 / Chapter 4.2.3 --- Identification of Reticulocytes --- p.148 / Chapter 4.2.4 --- Separation of Reticulocytes from erythrocytes --- p.149 / Chapter 4.2.5 --- Treatment of guinea pigs --- p.150 / Chapter 4.2.6 --- Oxygen Consumption --- p.150 / Chapter 4.2.7 --- Determination of Number of OBS --- p.154 / Chapter 4.2.8 --- STATISTICS --- p.154 / Chapter 4.3 --- RESULTS --- p.155 / Chapter 4.3.1 --- Establishment of the Method for Determinating the Number of OBS --- p.155 / Chapter 4.3.2 --- Scatchard Analysis of Binding of Ouabain to Erythrocytes --- p.155 / Chapter 4.3.3 --- Induction of Reticulocytosis and Treatment of animals --- p.160 / Chapter 4.3.4 --- Weight Loss and O2 Consumption in Control and T3 Treated Guinea Pigs --- p.160 / Chapter 4.3.5 --- Determination of OBS and Kd of Reticulocytes from Guinea Pigs Treated with T3 and Control --- p.164 / Chapter 4.4 --- DISCUSSION --- p.168 / Chapter CHAPTER 5 --- "THYROID HORMONES AND NA+,K+- ATP ASE ACTIVITY OF ERYTHROID CELLS" --- p.171 / Chapter 5.1 --- Review of the Literature --- p.172 / Chapter 5.1.1 --- Erythroid Differentiation and Maturation --- p.172 / Chapter 5.1.2 --- Methods of Separation of Erythroid Cells --- p.178 / Chapter 5.1.3 --- Biochemical Changes During Erythropoiesis --- p.179 / Chapter 5.1.4 --- In vitro Models of Differentiation and the Biochemical Changes During Haemoglobin Synthesis --- p.182 / Chapter 5.1.5 --- Aim of the Study --- p.185 / Chapter 5.2 --- MATERIALS & METHODS --- p.186 / Chapter 5.2.1 --- Materials --- p.186 / Chapter 5.2.2 --- Sterilisation of glassware --- p.186 / Chapter 5.2.3 --- Solutions --- p.186 / Chapter 5.2.4 --- Heat Inactivation of Foetal Bovine Serum --- p.187 / Chapter 5.2.4.1 --- Preparation of Cell Culture Medium (CCM) --- p.187 / Chapter 5.2.4.2 --- Sterility Test of CCM --- p.188 / Chapter 5.2.5 --- Growth of K562 cells --- p.188 / Chapter 5.2.5.1 --- Assessment of Cell Viability --- p.189 / Chapter 5.2.5.2 --- Subculture of Cells --- p.190 / Chapter 5.2.6 --- "Effect of T3 on Na+,K+-ATPase activity of K562 cell line" --- p.190 / Chapter 5.2.7 --- Induction of Haemoglobin Synthesis --- p.191 / Chapter 5.2.8 --- Benzidine Staining for Haemoglobin --- p.192 / Chapter 5.2.9 --- "The Determination of Na+,K+-ATPase activity" --- p.192 / Chapter 5.2.10 --- Determination of Protein --- p.194 / Chapter 5.3 --- RESULTS --- p.195 / Chapter 5.3.1 --- "Effect of Deoxycholate concentration on unmasking Na+,K+-ATPase activity" --- p.195 / Chapter 5.3.2 --- "Effect of varying saponin concentration on Na+, K+-ATPase activity" --- p.195 / Chapter 5.3.3 --- "Effect of Varying Incubation Time on Na+,K+-ATPase activity" --- p.198 / Chapter 5.3.4 --- "Effect of Varying the Amount of Protein on Na+,K+-ATPase activity" --- p.198 / Chapter 5.3.5 --- "Effect of varying ouabain concentration on Na+,K+-ATPase activity" --- p.201 / Chapter 5.3.6 --- Intra- and Inter-Assay Precision --- p.201 / Chapter 5.3.7 --- "Effect of T3 on the Na+,K+-ATPase Activity of K562 Cells" --- p.201 / Chapter 5.3.8 --- "T3 Concentration-Response Relationship for Na+,K+-ATPase Activity of K562 Cells" --- p.204 / Chapter 5.3.9 --- "Time course of Effect of T3 on Na+,K+-ATPase Activity of K562 cell line" --- p.207 / Chapter 5.3.10 --- Induction of Haemoglobin Synthesis in K562 cells --- p.210 / Chapter 5.3.11 --- Effect of Differentiation on T3 Stimulated K562 Cells --- p.210 / Chapter 5.4 --- DISCUSSION --- p.212 / Chapter CHAPTER 6 --- THYROID HORMONES AND THE ATP-DEPENDANT PROTEOLYTIC SYSTEM --- p.217 / Chapter 6.1 --- The ATP-Dependant Proteolytic System --- p.218 / Chapter 6.1.1 --- The Ubiquitin Pathway --- p.219 / Chapter 6.1.2 --- The Non-Ubiquitin Pathway --- p.222 / Chapter 6.1.3 --- ATP-dependant Proteolysis and Erythropoiesis --- p.223 / Chapter 6.1.4 --- Methods Used for Determining Proteolysis --- p.226 / Chapter 6.1.5 --- Aim of the Study --- p.227 / Chapter 6.2 --- MATERIALS & METHODS --- p.228 / Chapter 6.2.1 --- Materials --- p.228 / Chapter 6.2.2 --- Induction of Reticulocytosis and T3 Treatment --- p.228 / Chapter 6.2.3 --- Culture of K562 Cells and Effect of T3 --- p.229 / Chapter 6.2.4 --- Preparation of K562 Cell Lysate --- p.229 / Chapter 6.2.5 --- Preparation of Reticulocyte Extracts --- p.229 / Chapter 6.2.6 --- Iodination of Lysozyme --- p.230 / Chapter 6.2.7 --- Determination of Proteolytic Activity --- p.231 / Chapter 6.3 --- RESULTS --- p.234 / Chapter 6.3.1 --- Proteolytic Activity of Reticulocyte and K562 Cell Lysates --- p.234 / Chapter 6.3.2 --- Induction and Separation of Reticulocytes --- p.234 / Chapter 6.3.3 --- Effect of T3 Treatment on Guinea Pigs --- p.234 / Chapter 6.3.4 --- ATP-dependant proteolytic system of Reticulocytes and K562 Cells Treated with T3 and Controls --- p.239 / Chapter 6.4 --- DISCUSSION --- p.241 / Chapter CHAPTER 7 --- OVERVIEW & FUTURE WORK --- p.244 / Chapter 7.1 --- Overview & Future Work --- p.245 / REFERENCES --- p.249
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318649 |
| Date | January 1990 |
| Contributors | Arumanayagam, Mano., Chinese University of Hong Kong Graduate School. Division of Clinical and Pathological 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, 265 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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