Este trabalho avalia a influência de veículos de grande porte na capacidade de interseções rodoviárias em nível não semaforizadas, verificando as variações no comportamento de caminhões e ônibus durante manobras de cruzamento, em função de seu comprimento e de sua relação potência/peso. Foram realizadas análises das distribuições dos tempos de cruzamento e da aceitação de \"gaps\" por diferentes tipos de veículos, sendo utilizados modelos Logit para representar o comportamento desses veículos nos cruzamentos. Estes modelos serviram como base na elaboração de um simulador, para determinar a capacidade potencial das interseções e para calcular os fatores de conversão de veículos de grande porte em veículos de passageiro. / This study assesses the influence of heavy vehicles on the capacity of unsignalized rural intersections. This is done through the study of vehicle behavior during crossing maneuvers that can be attributed to length and power-to-weight ratio. Crossing time distributions and gap acceptance of diferent types of vehicles were analysed, and Logit models were used to represent their behavior. These models and a simulation model were used to determine the potencial capacity of intersections and to calculate factors to convert heavy vehicles into passenger vehicles.

by Xin-xia Tian. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 195-212). / 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.

by Kwan, Leo. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 108-116). / Abstract --- p.i / Achnowledgment --- p.ii / List of abbreviations --- p.iii / List of table and figures --- p.v / Table of contents --- p.vi / Chapter CHAPTER 1 --- INTRODUCTION / Chapter 1.1 --- THE DISCOVERY OF TUMOUR NECROSIS FACTOR (TNF) --- p.1 / Chapter 1.2 --- THE MOLECULE AND ITS RECEPTORS --- p.1 / Chapter 1.3 --- THE BIOLOGICAL ACTIVITIES OF TNF --- p.3 / Chapter 1.4 --- STUDIES ON THE CYTOTOXIC MECHANISM OF TNF --- p.4 / Chapter 1.5 --- A TENTATIVE MECHANISM OF TNF CYTOTOXICITY --- p.11 / Chapter 1.6 --- THE GLUTATHIONE SYSTEM : A CELLULAR PROTECTIVE MECHANISM AGAINST OXIDATIVE STRESS …… --- p.12 / Chapter 1.7 --- OBJECTIVE AND STRATEGY OF THIS STUDY --- p.16 / Chapter CHAPTER 2 --- MATERIALS AND APPARATI / Chapter 2.1 --- CELL LINES --- p.19 / Chapter 2.2 --- "ISOLATION, MAINTENANCE AND SUBCULTURE OF CELL LINES" --- p.19 / Chapter 1. --- Plain RPMI-1640 medium / Chapter 2. --- Penicillin-streptomycin solution / Chapter 3. --- Foetal bovine serum / Chapter 4. --- Complete RPMI-1640 medium / Chapter 5. --- Trypsin-ethylenediaminetetraacetate solution / Chapter 6. --- Phosphate buffered saline / Chapter 7. --- Cycloheximide / Chapter 8. --- Actinomycin D / Chapter 9. --- Trypan blue stain / Chapter 10. --- Neutral red stain / Chapter 11. --- Recombinant human tumour necrosis factor / Chapter 12. --- Cell culture plates and flasks / Chapter 2.3 --- GROWTH CHARACTERISTIC --- p.22 / Chapter 1. --- Tritiated Thymidine / Chapter 2. --- Tritiated Leucine / Chapter 3. --- Trichloroacetic acid / Chapter 4. --- Scintillation cocktail / Chapter 2.4 --- "RESPONSE TOWARDS ANTICANCER DRUGS, CYTOTOXIC AGENTS, AND ENZYME MODULATORS" --- p.23 / Chapter 1. --- N-acetyl-DL-homocysteinethiolactone / Chapter 2. --- Diethyldithiocarbamic acid / Chapter 3. --- Doxorubicin / Chapter 4. --- Acivicin / Chapter 5. --- Ethacrynic acid / Chapter 6. --- "L'Buthionine-[S,R]-sulfoximine" / Chapter 7. --- Hydrogen peroxide / Chapter 8. --- Methotrexate / Chapter 9. --- Menadione / Chapter 2.5 --- CULTURE OF BACTERIAL CELLS --- p.27 / Chapter 1. --- Ampicillin stock solution / Chapter 2. --- Chloramphenicol stock solution / Chapter 3. --- Tetracycline stock solution / Chapter 4. --- Luria-Bertani medium / Chapter 5. --- LB with ampicillin / Chapter 6. --- SOB medium / Chapter 7. --- SOB medium with ampicillin / Chapter 8. --- SOC medium / Chapter 9. --- SB medium / Chapter 10. --- SB medium with ampicillin / Chapter 11. --- Agar plates / Chapter 2.6 --- PREPARATION OF DNA PROBES FROM BACTERIAL CLONES --- p.29 / Chapter 1. --- FlexiPrep Kit / Chapter 2. --- Restriction endonucleases / Chapter 3. --- GeneClean® II Kit / Chapter 4. --- cDNA clones for making DNA probes / Chapter 5. --- TrisHCl EDTA buffer / Chapter 2.7. --- ELECTROPHORESIS OF DNA --- p.31 / Chapter 1. --- EDTA stock solution / Chapter 2. --- Tris acetate EDTA electrophoresis buffer / Chapter 3. --- Tris borate EDTA electrophoresis buffer / Chapter 4. --- Ethidium bromide / Chapter 5. --- DNA molecular size markers / Chapter 6. --- TAE/TBE agarose gel slab / Chapter 2.8 --- CONSTRUCTION OF MURINE TNFR1 PARTIAL cDNA CLONE --- p.33 / Chapter 1. --- Frist strand cDNA synthesis Kit / Chapter 2. --- Murine TNFR1 forward and reverse primers / Chapter 3. --- Polymerase chain reaction reagents / Chapter 4. --- Cloning vector / Chapter 5. --- Modifing enzymes / Chapter 6. --- T7 SequencingTM Kit / Chapter 7. --- Acrylamide/bis gel stock solution / Chapter 8. --- Urea / Chapter 9. --- TEMED and ammonium persulphate / Chapter 10. --- β-Galactosidase colour test reagents / Chapter 11. --- TFB solution / Chapter 12. --- DnD solution / Chapter 2.9. --- RADIOLABELLING OF DNA PROBES --- p.35 / Chapter 1. --- Oligolabelling kit / Chapter 2. --- Redivue [α-32P] dCTP / Chapter 3. --- PUSH column / Chapter 2.10 --- EXTRACTION OF TOTAL RNA FROM CELL LINES --- p.36 / Chapter 1. --- N-Lauroylsarcosine / Chapter 2. --- 2M sodium acetate (pH48) / Chapter 3. --- Phenol / Chapter 4. --- Isopropanol / Chapter 5. --- Ethanol / Chapter 6. --- Extraction buffer / Chapter 7. --- Chloroform / Chapter 8. --- Isoamyl alcohol / Chapter 2.11 --- HYBRIDIZATION AND NORTHERN ANALYSIS --- p.37 / Chapter 1. --- 20XSSC / Chapter 2. --- 5X formaldehyde running buffer / Chapter 3. --- RNA sample buffer / Chapter 4. --- 10X RNA loading buffer / Chapter 5. --- Formaldehyde slab gel / Chapter 6. --- Hybond®-N membrane / Chapter 7. --- Immobilon®-N membrane / Chapter 8. --- Salmon sperm DNA / Chapter 9. --- Sodium dodecyl sulphate / Chapter 10. --- Dextran sulphate / Chapter 11. --- Kodak Biomax MR and X-OMAT films and developing kits / Chapter 2.12 --- APPARATI USED --- p.39 / Chapter CHAPTER 3 --- METHODS / Chapter 3.1 --- ISOLATION AND MAINTENANCE OF TNF RESISTANT L929 CELLS --- p.40 / Chapter 3.1.1 --- Culture of L929 cells / Chapter 3.1.2 --- Trypan blue exclusion test / Chapter 3.1.3 --- Isolation of TNF-resistant variants of L929 / Chapter 3.1.4 --- Verification of the TNF-resistant trait of rL929 / Chapter 3.1.5 --- Neutral red uptake assay / Chapter 3.2 --- COMPARING L929 AND rL929 CELLS IN TERMS OF GROWTH CHARACTERISTICS --- p.43 / Chapter 3.2.1 --- Doubling time / Chapter 3.2.2 --- Rate of protein synthesis / Chapter 3.2.3 --- Rate of DNA synthesis / Chapter 3.3 --- COMPARING L929 AND rL929 CELLS IN TERMS OF RESPONSE TOWARDS DIFFERENT ENZYME INHIBITORS AND CYTOTOXIC AGENTS --- p.44 / Chapter 3.3.1 --- TNF cytotoxicity on L929 and rL929 cells --- p.44 / Chapter 3.3.2 --- Effects of inhibitors of gene transcription and protein synthesis on TNF cytotoxicity on L929 and rL929 cells --- p.44 / Chapter 3.3.3 --- Cytotoxic effect of hydrogen peroxide and menadione on L929 and rL929 cells --- p.44 / Chapter 3.3.4 --- TNF cytotoxicity on L929 and rL929 cells: effect of N-acetyl homocysteine thiolatone --- p.45 / Chapter 3.3.4.1 --- The tolerant limit of AHCT / Chapter 3.3.4.2 --- Effect of AHCT on TNF cytotoxicity / Chapter 3.3.5 --- TNF cytotoxicity on L929 and rL929 cells: effect of diethyldithiocarbamate --- p.46 / Chapter 3.3.5.1 --- The tolerant limit of DEDTC / Chapter 3.3.5.2 --- Effect of DEDTC on TNF cytotoxicity / Chapter 3.3.6 --- TNF cytotoxicity on L929 and rL929 cells: effect of buthionice sulfoximine --- p.47 / Chapter 3.3.6.1 --- The tolerant limit of BSO / Chapter 3.3.6.2 --- Effect of BSO on TNF cytotoxicity / Chapter 3.3.7 --- TNF cytotoxicity on L929 and rL929 cells: effect of Acivicin --- p.47 / Chapter 3.3.7.1 --- The tolerant limit of acivicin / Chapter 3.3.7.2 --- Effect of acivicin on TNF cytotoxicity / Chapter 3.3.8 --- TNF cytotoxicity on L929 and rL929 cells: effect of ethacrynic acid --- p.48 / Chapter 3.3.8.1 --- The tolerant limit of ethacrynic acid / Chapter 3.3.8.2 --- Effect of ethacrynic acid on TNF cytotoxicity / Chapter 3.3.9 --- Cytotoxic effect of doxorubicin on L929 and rL929 cells --- p.49 / Chapter 3.3.10 --- TNF cytotoxicity of L929 cells: effect of N-acetyl cysteine --- p.49 / Chapter 3.3.11 --- Cytotoxic effect of methotrexate on L929 and rL929 cells --- p.50 / Chapter 3.3.12 --- Cytotoxic effect of hyperthermia on L929 and rL929 cells --- p.50 / Chapter 3.4 --- NORTHERN ANALYSIS AND HYBRIDIZATION --- p.51 / Chapter 3.4.1. --- Preparing RNA blots --- p.51 / Chapter 3.4.1.1 --- Extraction of total RNA from cells / Chapter 3.4.1.2 --- Making equal loading of RNA samples in formaldehyde gel electrophoresis / Chapter 3.4.1.3 --- Northern blotting of RNA / Chapter 3.4.2. --- Preparation of cDNA probes --- p.53 / Chapter 3.4.2.1 --- Preparing plasmids from A TCC clones / Chapter 3.4.2.2 --- Preparing TNFR1 probe from first-strand cDNA of L929 cells / Chapter 1. --- Construction of recombinant clone from the PCR product of TNFRl fragment / Chapter 2. --- Transforming the recombinant vector into JM109 host / Chapter 3. --- Sequencing of PCR product for identity confirmation / Chapter 3.4.2.3 --- Preparing DNA inserts from plasmids / Chapter 3.4.3 --- Radiolabelling of cDNA probes --- p.56 / Chapter 3.4.4 --- Hybridization of radioactive probes to RNA blots --- p.57 / Chapter CHAPTER 4 --- RESULTS AND DISCUSSIONS / Chapter 4.1 --- ISOLATION OF TNF-RESISTANT VARIANTS OF L929 CELLS --- p.58 / Chapter 4.1.1 --- Single cell subcloning of TNF-resistant L929 variants / Chapter 4.1.2 --- Growth rates of L929 and rL929 cells / Chapter 4.1.3 --- Rate of protein synthesis in L929 and rL929 cells / Chapter 4.1.4 --- Rate of DNA synthesis in L929and rL929 cells / Chapter 4.2 --- EFFECT OF INHIBITORS OF GENE TRANSCRIPTION AND PROTEIN SYNTHESIS ON TNF CYTOTOXICITY ON L929 AND rL929 CELLS --- p.67 / Chapter 4.3 --- RESPONSE OF L929 AND rL929 CELLS TOWARDS VARIOUS CYTOTOXIC AGENTS --- p.70 / Chapter 4.3.1 --- "Response towards methotrexate, an anti-metabolite used in cancer treatment" / Chapter 4.3.2 --- "Response towards doxorubicin, an chemotherapeutic agent used in cancer treatment" / Chapter 4.3.3 --- "Response towards menadione, a cytotoxic agent known to generate free radicals inside cells" / Chapter 4.3.4 --- Response towards hydrogen peroxide: a highly oxidative agent / Chapter 4.3.5 --- "Response towards hyperthermia, a treatment known to exert oxidative stress on cells" / Chapter 4.4 --- EFFECTS OF MODULATORS OF CYTOSOLIC SUPEROXIDE DISMUTASE ON TNF CYTOTOXICITY ON L929 and rL929 CELLS --- p.77 / Chapter 4.5 --- EFFECT OF MODULATORS OF GLUTATHIONE METABOLISM ON TNF CYTOTOXICITY ON L929 AND rL929 CELLS --- p.82 / Chapter 4.5.1 --- "Effects of L-buthionine [S,R] sulfoximine, an inhibitor of glutathione synthesis" --- p.82 / Chapter 4.5.2 --- "Effect of N-acetyl cysteine, a cysteine derivative" --- p.84 / Chapter 4.5.3 --- "Effects of acivicin , an inhibitor of GSH reuptake and recycle" --- p.85 / Chapter 4.5.4 --- "Effect of ethacrynic acid, an inhibitor of glutathione S- transferase" --- p.87 / Chapter 4.6 --- GENE EXPRESSION IN L929 AND rL929 CELLS IN THE COURSE OF TNF CHALLENGE --- p.89 / Chapter 4.6.1 --- Isolation of total RNA from L929 and rL929 cells --- p.89 / Chapter 4.6.2 --- Preparation of DNA probes for hybridization --- p.89 / Chapter 4.6.3 --- Hybridization of specific probes on RNA blots --- p.90 / Chapter 4.6.3.1 --- Expression of heat shock protein --- p.70 / Chapter 4.6.3.2 --- Expression of the p55 TNF receptor / Chapter 4.6.3.3 --- Expression of glutathione reductase / Chapter 4.6.3.4 --- Expression of glutathione S-transferase pi / Chapter 4.7 --- DISCUSSIONS OF THE EXPERIMENTAL RESULTS --- p.97 / Chapter CHAPTER 5 --- GENERAL DISCUSSION --- p.104 / APPENDIX / Generation of the TNF receptor 1 cDNA probe --- p.106 / REFERENCES --- p.108

Ho, Sui Fan Tong. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1992. / Includes bibliographical references (leaves 68-71). / LIST OF TABLES / LIST OF FIGURES / ACKNOWLEDGEMENTS / ABSTRACT / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 2. --- OBJECTIVES --- p.12 / Chapter 3. --- MATERIALS AND METHODS --- p.13 / Chapter 3.1 --- Materials --- p.13 / Chapter 3.1.1 --- Enzymes --- p.13 / Chapter 3.1.2 --- DNA Markers --- p.13 / Chapter 3.1.3 --- General Reagents --- p.13 / Chapter 3.2 --- General Methods --- p.15 / Chapter 3.2.1 --- Phenol and Phenol/Chloroform (1:1) Preparation --- p.15 / Chapter 3.2.2 --- Buffer Preparation --- p.15 / Chapter 3.2.3 --- Agarose Gel Electrophoresis --- p.18 / Chapter 3.2.4 --- Polyacrylamide Gel Electrophoresis --- p.18 / Chapter 3.3 --- DNA Study --- p.19 / Chapter 3.3.1 --- Haemophilia B Patient --- p.19 / Chapter 3.3.2 --- Blood Collection --- p.20 / Chapter 3.3.3 --- DNA Extraction --- p.20 / Chapter 3.3.4 --- DNA Quantitation --- p.21 / Chapter 3.3.5 --- Polymerase Chain Reaction --- p.22 / Chapter 3.3.6 --- Purification of PCR Products --- p.28 / Chapter 3.3.7 --- Sequencing --- p.32 / Chapter 3.3.8 --- Cloning --- p.37 / Chapter 4. --- RESULTS --- p.40 / Chapter 4.1 --- DNA Extraction --- p.40 / Chapter 4.2 --- Calibration of the Coy TempCycler --- p.42 / Chapter 4.3 --- Optimization of PCR --- p.44 / Chapter 4.3.1 --- PCR-1 --- p.44 / Chapter 4.3.2 --- PCR-2 --- p.46 / Chapter 4.3.3 --- PCR-3 --- p.46 / Chapter 4.3.4 --- PCR-4 --- p.48 / Chapter 4.3.5 --- PCR-5 --- p.49 / Chapter 4.3.6 --- PCR-6 --- p.50 / Chapter 4.3.7 --- PCR-7 --- p.51 / Chapter 4.4 --- Purification of PCR Product --- p.52 / Chapter 4.4.1 --- GC-1 --- p.52 / Chapter 4.4.2 --- GC-2 --- p.52 / Chapter 4.4.3 --- GC-3 --- p.53 / Chapter 4.4.4 --- PAGE-1 --- p.54 / Chapter 4.4.5 --- PAGE-2 --- p.54 / Chapter 4.4.6 --- Agarose Gel Extraction with Glasswool Exclusion --- p.55 / Chapter 4.5 --- Direct Sequencing of PCR Products --- p.55 / Chapter 4.6 --- Cloning --- p.55 / Chapter 5. --- DISCUSSION --- p.57 / Chapter 5.1 --- DNA Extraction --- p.57 / Chapter 5.2 --- Polymerase Chain Reaction --- p.57 / Chapter 5.3 --- Purification of PCR Products --- p.58 / Chapter 5.4 --- Sequencing --- p.61 / Chapter 5.5 --- Cloning --- p.61 / Chapter 6. --- CONCLUSION --- p.67 / Chapter 7. --- PHOTOGRAPHS --- p.64 / Chapter 8. --- REFERENCES --- p.68

by Suk-Fung Tsang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 101-112). / Abstract --- p.i / Acknowledgement --- p.iii / Abbreviation --- p.iv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- TNF molecule / Chapter 1.2 --- Molecular biosynthesis of TNF / Chapter 1.3 --- Antitumor activity of TNF / Chapter 1.4 --- Macrophage-mediated immunity / Chapter 1.5 --- Endogenous production of TNF / Chapter 1.6 --- LPS : the potent inducer for TNF release / Chapter 1.7 --- Natural product: as primer or inducer / Chapter 1.8 --- Aim of this project / Chapter Chapter 2 --- Materials and Methods --- p.22 / Chapter 2.1 --- Materials / Chapter 2.2 --- Animals / Chapter 2.3 --- Cell line / Chapter 2.4 --- Transformed cell line : EAT cells invivo / Chapter 2.5 --- Reagents / Chapter 2.6 --- Methods / Chapter Chapter 3 --- Preparation of sample --- p.33 / Chapter 3.1 --- Alcohol precipitaion of Bupleuri radix / Chapter 3.2 --- Endogenous TNF production by BR fractions / Chapter Chapter 4 --- Purification of BRI --- p.38 / Chapter 4.1 --- Gel filtration chromatography of BRI / Chapter 4.2 --- Anion exchange chromatography / Chapter Chapter 5 --- Purification of PQI --- p.52 / Chapter 5.1 --- Gel filtration chromatography of PQI / Chapter 5.2 --- Anion exchange chromatography / Chapter Chapter 6 --- Capacity of BR and PQ as eliciting agent for endogenous TNF production --- p.62 / Chapter 6.1 --- Time course of endogenous TNF production by BRI subfractions / Chapter 6.2 --- Time course of endogenous TNF production by PQI subfractions / Chapter 6.3 --- BRI subfractions as eliciting agents / Chapter 6.4 --- PQI subfractions as eliciting agents / Chapter Chapter 7 --- Are BR and PQ priming agents in endogenous TNF production ？ --- p.71 / Chapter 7.1 --- Priming by intraperitoneal route / Chapter 7.2 --- Priming by intravenous route / Chapter Chapter 8 --- Removal of LPS by acetic acid treatment --- p.79 / Chapter Chapter 9 --- Antitumor activities of BRI subfractionsin relationship with TNF production --- p.86 / Chapter 9.1 --- BRI subfraction as eliciting agent / Chapter 9.2 --- Pretreatment with BRIA subfractions followed by LPS treatment / Chapter Chapter 10 --- Conclusion --- p.95 / Bibliography --- p.101

Ringo M.H. Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 75-81). / ACKNOWLEDGMENT --- p.2 / ABSTRACT --- p.3 / TABLE OF CONTENTS --- p.5 / LIST OF TABLES --- p.8 / Chapter CHAPTER 1 --- PREVIOUS STUDIES ON USING PROCRUSTES ROTATION TO ASSESS FACTORIAL INVARIANCE --- p.10 / Factorial invariance problem --- p.10 / Procrustes rotation with congruent coefficient as a way to test factorial invariance --- p.11 / Quantifying the Procrustes fit --- p.14 / Outline of the present study --- p.15 / Chapter CHAPTER 2 --- A CRITICAL EVALUATION OF THE PERMUTATION METHOD --- p.18 / Introduction --- p.18 / Method --- p.19 / Results and Discussions --- p.21 / Chapter CHAPTER 3 --- BOOTSTRAP TESTING PROCEDURE FOR A FULLY SPECIFIED TARGET --- p.24 / Introduction --- p.24 / A brief introduction to the bootstrap procedure --- p.24 / The bootstrap testing procedure for a fully specified target --- p.26 / Method --- p.28 / Results and Discussions --- p.28 / Chapter CHAPTER 4 --- BOOTSTRAP TESTING FOR A PARTIALLY SPECIFIED TARGET --- p.33 / Introduction --- p.33 / The bootstrap testing procedure for a partially specified target --- p.36 / Method --- p.38 / Quantifying the fit - congruence coefficients for the partial target rotation --- p.39 / Results and Discussions --- p.40 / Chapter CHAPTER 5 --- FURTHER EXTENSIONS OF THE BOOTSTRAP METHOD --- p.45 / Introduction --- p.45 / First extension - when correlation matrix is used --- p.45 / The modified bootstrap procedure --- p.45 / Method --- p.48 / Results and Discussions --- p.48 / Second extension - when raw data of the target sample is not available --- p.49 / The conditional bootstrap procedure for a fully specified target --- p.49 / Method --- p.50 / Results and Discussions --- p.51 / Chapter CHAPTER 6 --- THREE REAL EXAMPLES --- p.54 / Example 1 - Testing factorial invariance of CPAI between two random split samples --- p.54 / Results --- p.55 / Example 2 - Testing factorial invariance of CPAI between Chinese males and females --- p.56 / Results --- p.57 / Example 3 - Cross-cultural comparison of the Big Five Model between U. S. and Chinese samples --- p.58 / Results --- p.59 / Chapter CHAPTER 7 --- CONCLUSIONS --- p.62 / Practical remarks on the bootstrap procedure --- p.62 / A note on the transformation on the sample for constructing correct resampling space --- p.64 / Remarks on utilizing the congruence coefficients --- p.65 / How good are the congruence coefficients in detecting discrepancy between two factor structures? --- p.68 / Rule of thumb for factor congruence coefficient in checking factor replicability --- p.68 / Sample size requirement --- p.69 / Limitations of the present study --- p.70 / Direction of future studies --- p.71 / Concluding remarks --- p.73 / REFERENCES --- p.75 / NOTES --- p.82 / APPENDIX1 --- p.83 / TABLES 1 TO TABLES17 --- p.84

Tse Chui-ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 130-146). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / 論文撮要 --- p.iii / List of Figures and Tables --- p.iv / Abbreviations --- p.vi / Table of contents --- p.vii / Chapter Chapter I --- Introduction --- p.1 / Chapter 1.1 --- Literature review --- p.1 / Chapter 1.1.1 --- An overview of IGFs --- p.3 / Chapter 1.1.2 --- Molecular biology of IGFs --- p.5 / Chapter 1.1.2.1 --- IGF-I and IGF-II genes and mRNAs --- p.5 / Chapter 1.1.2.2 --- Amino acid sequences of IGF-II --- p.8 / Chapter 1.1.2.3 --- Imprinting of IGF-II --- p.12 / Chapter 1.1.3 --- IGF distribution in tissues and body fluids --- p.14 / Chapter 1.1.3.1 --- IGF in serum --- p.14 / Chapter 1.1.3.2 --- IGF binding proteins --- p.16 / Chapter 1.1.4 --- IGF receptors --- p.19 / Chapter 1.1.4.1 --- Structures of the IGF receptors --- p.20 / Chapter 1.1.4.2 --- Ligand binding of the IGF receptors --- p.21 / Chapter 1.1.4.3 --- Signal transduction and biological response --- p.22 / Chapter 1.1.5 --- Biological effects of IGF --- p.24 / Chapter 1.1.6 --- Expression of recombinant IGF --- p.28 / Chapter 1.2 --- Rationale and Objective --- p.29 / Chapter Chapter II --- Methodology --- p.33 / Chapter 2.1 --- Design of degenerate primers --- p.33 / Chapter 2.2 --- Cloning --- p.35 / Chapter 2.2.1 --- DNA extraction from agarose gel --- p.35 / Chapter 2.2.2 --- Linearization and dephosphorylation of plasmid DNA --- p.35 / Chapter 2.2.3 --- Blunt-end ligation of amplicon with linearized plasmid --- p.36 / Chapter 2.2.4 --- T/A ligation of amplicon with linearized plasmid --- p.37 / Chapter 2.2.5 --- Sticky end ligation of foreign DNA with linearized plasmid --- p.37 / Chapter 2.2.6 --- Preparation of competent of E. coli stain DHI5α cells --- p.38 / Chapter 2.2.7 --- Transformation of plasmid vector into competent cells (heat-shock/ electroporation) --- p.39 / Chapter 2.2.8 --- "Spread single colony, PCR check clone and inoculation" --- p.40 / Chapter 2.2.9 --- Small scale alkali preparation of plasmid DNA --- p.41 / Chapter 2.2.10 --- Large scale preparation of plasmid DNA --- p.41 / Chapter 2.2.11 --- Nucleotide sequencing --- p.41 / Chapter 2.2.11.1 --- Manual sequencing --- p.41 / Chapter 2.2.11.2 --- PCR sequencing --- p.43 / Chapter 2.3 --- Northern blot --- p.45 / Chapter 2.4 --- Preparation of radio-labeled probe and hybridization of radio-labeled probe to nylon immobilized nucleic acid --- p.46 / Chapter 2.5 --- RACE --- p.48 / Chapter 2.5.1 --- Design of gene-specific primer --- p.51 / Chapter 2.5.2 --- First strand cDNA synthesis --- p.51 / Chapter 2.5.3 --- TdT tailing of cDNA --- p.52 / Chapter 2.6 --- Poly-A tract extraction --- p.53 / Chapter 2.7 --- Tissue distribution of mRNA --- p.53 / Chapter 2.7.1 --- Tissue preparation --- p.53 / Chapter 2.7.2 --- Total RNA extraction --- p.54 / Chapter 2.7.3 --- Formaldehyde agarose gel electrophoresis of RNA --- p.54 / Chapter 2.8 --- RNAse protection assay --- p.55 / Chapter 2.8.1 --- Antisense probe generation --- p.56 / Chapter 2.8.2 --- Preparation of the sample RNA --- p.58 / Chapter 2.8.3 --- Hybridization --- p.58 / Chapter 2.8.4 --- RNase digestion of hybridized probe and sample RNA --- p.59 / Chapter 2.8.5 --- Preparation of radioactive marker --- p.60 / Chapter 2.8.6 --- Separation and detection of protected fragments --- p.60 / Chapter 2.8.7 --- Data processing and statistical analysis --- p.61 / Chapter 2.9 --- Injection of GH --- p.62 / Chapter 2.10 --- Recombinant protein expression --- p.62 / Chapter 2.10.1 --- Plasmid construction --- p.62 / Chapter 2.10.2 --- Expression --- p.63 / Chapter 2.11 --- Resolution of proteins on SDS-PAGE --- p.63 / Chapter 2.12 --- Purification --- p.64 / Chapter 2.13 --- Western transfer --- p.64 / Chapter 2.14 --- Immunodetection --- p.65 / Chapter Chapter III --- Results & Discussion --- p.67 / Chapter 3.1 --- Isolation and characterization of IGF-II cDNA and its gene organization --- p.67 / Chapter 3.1.1 --- Introduction --- p.67 / Chapter 3.1.2 --- Results --- p.68 / Chapter 3.1.2.1 --- Generation of a fragment of the common carp IGF-II cDNA by PCR --- p.68 / Chapter 3.1.2.2 --- Isolation of the full length common carp IGF-II cDNA by RACE. --- p.69 / Chapter 3.1.2.3 --- Nucleotide sequence analysis --- p.74 / Chapter 3.1.2.4 --- Relationship of common carp IGF-II to common carp IGF-I and insulin --- p.78 / Chapter 3.1.2.5 --- Confirmation of the presence of IGF-II in common carp --- p.79 / Chapter 3.1.2.6 --- Multiple mRNA forms of common carp IGF-I and IGF-II --- p.80 / Chapter 3.1.2.7 --- Gene organization of the common carp IGF-II gene --- p.83 / Chapter 3.1.3 --- Discussion --- p.86 / Chapter 3.2 --- Tissue specific distribution of IGF-I and IGF-II mRNA and their hormonal regulation --- p.90 / Chapter 3.2.1 --- Introduction --- p.90 / Chapter 3.2.2 --- Results --- p.94 / Chapter 3.2.2.1 --- RNase protection assay measurement of tissue mRNA levels in juvenile and adult common carp --- p.94 / Chapter 3.2.2.2 --- Expression of IGF-II mRNA during larval development --- p.99 / Chapter 3.2.2.3 --- Effect of GH on IGF-I and IGF-II mRNA levels in brain and Hver of juvenile common carp --- p.102 / Chapter 3.2.3 --- Discussion --- p.106 / Chapter 3.3 --- Recombinant common carp IGF-II expressed in E. coli --- p.110 / Chapter 3.3.1 --- Introduction --- p.110 / Chapter 3.3.2 --- Results --- p.112 / Chapter 3.3.2.1 --- Product of recombinant common carp IGF-II --- p.112 / Chapter 3.3.2.2 --- Purification of common carp IGF-II --- p.115 / Chapter 3.3.2.3 --- Immunodetection --- p.117 / Chapter 3.3.3 --- Discussion --- p.118 / Chapter Chapter IV --- General conclusion --- p.120 / Appendix: Reagents --- p.124 / Reference list --- p.130

Mutations in FGFR2 are common in a subset of endometrial carcinomas. Given the emergence of small molecule inhibitors specific to this receptor tyrosine kinase, FGFR2 is an attractive therapeutic target. However, compensatory and adaptation mechanisms limit the clinical utility of compounds that target nodes in the receptor tyrosine kinase network. Here, we analysed the impact of FGFR inhibition in endometrial cancer cells and observed the emergence of a resistant population in an FGFR2-mutant cell line. To understand the mechanisms underlying this adaptation response, we used a phosphoproteomics approach to measure the kinase network in an unbiased manner. These experiments led to the identification of an AKT-related compensatory mechanism underpinning this resistance. Further dissection of this resistance mechanism utilising gene expression analysis showed PHLDA1, a negative regulator of AKT, was significantly down-regulated in resistant cells. This was further confirmed at the protein level. siRNA knockdown of PHLDA1 conferred immediate drug resistance in the FGFR2-mutant endometrial cancer cell line. Therefore, we identified PHLDA1 down-regulation as a mediator of drug resistance in FGFR2 mutant cancer cells, the first demonstration of the role of PHLDA1 in the acquisition and maintenance of drug resistance. Using a 3D physiomimetic model, we demonstrated that AKT inhibition alone also led to generation of a drug-resistant population. Most importantly, dual-drug therapy inhibited proliferation and induced cell death. Our data highlight how mass spectrometry and microarray gene expression analysis can complement each other in the identification of novel resistance mechanisms in cancer cells. These data suggest that combination treatment of FGFR2-mutant endometrial cancers, targeting both FGFR2 and AKT, represents a promising therapeutic approach.

Laser Raman Spectroscopy (LRS) has been employed to characterise the structure of carbon fibres, the effect of surface treatment and the response of the material to externally applied loads. The strain sensitivity provided a unique relationship between the applied strain and the Raman frequency for each type of fibre, termed as the Raman Frequency Gauge Factor. After examining a wide range of fibres, of various Young's moduli and various manufacturing routes, it was concluded that both tensile and compressive properties of carbon fibres can be improved by controlling the fibre morphology during manufacture. This morphological control seems to achieve its objectives by reducing the skin-core effect in the fibre structure. The result of such an alteration can be detected in tension by the increase of the initial fibre modulus and in compression, by the absence of premature catastrophic type of failure. Nevertheless, non-linear stress-strain phenomena seem to be a permanent feature of all carbon fibres and the significant modulus softening in compression appears to determine the limits of the fibre compressive strength. The load transfer mechanism at the carbon fibre/epoxy resin interface has been subsequently investigated during the fibre fragmentation process in a single fibre model composite. The fibre strain distribution along the fibre fragments has been derived through the Raman spectrum of the fibre and its Raman Frequency Gauge Factor. In turn, the interfacial shear stress distribution has been evaluated using a simple balance of forces model. The maximum shear stress, allowed to develop at the f ibre/matrix interface, has been considered as a reasonable estimate of its interfacial strength. It was concluded that both the fibre surface treatment and the use of a lower modulus filament can increase the system's interfacial strength, reduce debonding propagation and withhold the interfacial yielding in the vicinity of the fibre discontinuities.

by Yuen Wai-ying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 50-51). / Abstract also in Chinese. / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Estimation of Factor Scores in a Three-level Factor Analysis Model / Chapter 2.1 --- The Three-level Factor Analysis Model --- p.5 / Chapter 2.2 --- Estimation of Factor Scores in Between-group --- p.7 / Chapter 2.2.1 --- REG Method --- p.9 / Chapter 2.2.2 --- GLS Method --- p.11 / Chapter 2.3 --- Estimation of Factor Scores in Second Level Within-group --- p.13 / Chapter 2.3.1 --- REG Method --- p.15 / Chapter 2.3.2 --- GLS Method --- p.17 / Chapter 2.4 --- Estimation of Factor Scores in First Level Within-group / Chapter 2.4.1 --- First Approach --- p.19 / Chapter 2.4.2 --- Second Approach --- p.24 / Chapter 2.4.3 --- Comparison of the Two Approaches in Estimating Factor Scores in First Level Within-group --- p.31 / Chapter 2.5 --- Summary on the REG and GLS Methods --- p.35 / Chapter Chapter 3 --- Simulation Studies / Example1 --- p.37 / Example2 --- p.42 / Chapter Chapter 4 --- Conclusion and Discussion --- p.48 / References --- p.50 / Figures --- p.52