Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

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

by Tung Kai Chiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 152-166). / Abstracts in English and Chinese. / STATEMENT --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABBREVIATIONS --- p.iv / ABSTRACT --- p.vii / 撮要 --- p.x / TABLE OF CONTENTS --- p.xiii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- AN OVERVIEW OF THE IMMUNE SYSTEM --- p.1 / Chapter 1.1.1 --- Innate Immunity --- p.1 / Chapter 1.1.2 --- Specific Acquired Immunity --- p.4 / Chapter 1.2 --- IMMUNE RESPONSE TO TUMOURS --- p.7 / Chapter 1.2.1 --- Specific Mechanisms --- p.7 / Chapter 1.2.1.1 --- Cellular Immune Response --- p.7 / Chapter 1.2.1.2 --- Humoral Immune Response --- p.9 / Chapter 1.2.2 --- Non-specific Mechanism --- p.10 / Chapter 1.2.2.1 --- Natural Killer Cells --- p.10 / Chapter 1.2.2.2 --- Lymphokine-activated Cells --- p.11 / Chapter 1.2.2.3 --- Tumour Infiltrating Lymphocytes --- p.12 / Chapter 1.2.2.4 --- Macrophages --- p.13 / Chapter 1.2.2.5 --- Other Cell Types Mediating Non-specific Immunity --- p.14 / Chapter 1.3 --- PHYTOCHEMICALS AS POTENTIAL IMMUNOMODULATORS and anti-tumour agents --- p.15 / Chapter 1.3.1 --- Modulation of the Immune System --- p.15 / Chapter 1.3.2 --- Induction of Cancer Cell Differentiation --- p.16 / Chapter 1.3.3 --- Stimulation of Apoptosis --- p.17 / Chapter 1.3.4 --- Other Anti-tumour Mechanisms --- p.18 / Chapter 1.4 --- GREEN TEA: GENERAL PROPERTIES AND PHARMACO- logical activities --- p.19 / Chapter 1.4.1 --- General Introduction --- p.19 / Chapter 1.4.2 --- Chemistry of Tea --- p.21 / Chapter 1.4.3 --- Physiological and Pharmacological Effects of Green Tea Catechins --- p.26 / Chapter 1.4.3.1 --- Anti-oxidative Activity --- p.26 / Chapter 1.4.3.2 --- Anti-mutagenic Activity --- p.27 / Chapter 1.4.3.3 --- Anti-carcinogenic Activity --- p.27 / Chapter 1.4.3.4 --- Anti-inflammatory Activity --- p.32 / Chapter 1.4.3.5 --- Hypocholesterolemic and Hypolipidemic Activities --- p.32 / Chapter 1.4.3.6 --- Anti-microbial Activity --- p.33 / Chapter 1.5 --- aims and scopes of this investigation --- p.34 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- MATERIALS --- p.36 / Chapter 2.1.1 --- Animals --- p.36 / Chapter 2.1.2 --- Cell lines --- p.36 / Chapter 2.1.3 --- Green Tea Catechins and Epicatechin Isomers --- p.38 / Chapter 2.1.4 --- Recombinant Murine Interleukin-2 (rmIL-2) --- p.38 / Chapter 2.1.5 --- Antibodies --- p.39 / Chapter 2.1.6 --- "Buffers, Culture Medium and Other Reagents" --- p.40 / Chapter 2.1.7 --- Reagents for Gel Electrophoresis --- p.47 / Chapter 2.1.8 --- Radioisotopes --- p.48 / Chapter 2.2 --- METHODS --- p.49 / Chapter 2.2.1 --- "Isolation, Purification and Characterization of Green Tea Catechins and Epicatechin Isomers" --- p.49 / Chapter 2.2.1.1 --- Extraction of Green Tea Catechins --- p.49 / Chapter 2.2.1.2 --- Analysis of Epicatechin Isomers by HPLC --- p.49 / Chapter 2.2.1.3 --- Determination of the Bio-toxicity of Green Tea Catechins --- p.50 / Chapter 2.2.1.4 --- Assay of In Vitro Cytotoxicity of Green Tea Catechins on Splenocytes --- p.51 / Chapter 2.2.2 --- Assays for the Immunomodulatory Activities of Green Tea Catechins --- p.52 / Chapter 2.2.2.1 --- Isolation and Preparation of Cells --- p.52 / Chapter 2.2.2.2 --- In Vitro Lymphocyte Transformation Assay --- p.53 / Chapter 2.2.2.3 --- Mixed Lymphocyte Culture --- p.53 / Chapter 2.2.2.4 --- Colorimetric Assay of Alloreactive Cytotoxic T Lymphocytes --- p.54 / Chapter 2.2.2.5 --- Foodpad Swelling Assay of Delayed-type Hypersensitivity --- p.55 / Chapter 2.2.2.6 --- Haemagglutination Assay of In Vivo Antibody Formation --- p.56 / Chapter 2.2.2.7 --- Characterization of the Lymphocyte Subpopulations from Spleen by Flow Cytometry --- p.56 / Chapter 2.2.2.8 --- In Vivo Migration of Macrophages --- p.57 / Chapter 2.2.2.9 --- In Vitro Assay of Phagocytic Activity of PEC --- p.58 / Chapter 2.2.2.10 --- In Vitro Assay of Macrophage-mediated Cytostatic Activity --- p.58 / Chapter 2.2.3 --- Assays for the Anti-tumour Activities of Green Tea Catechins --- p.60 / Chapter 2.2.3.1 --- Assay of In Vivo Anti-tumour Activity --- p.60 / Chapter 2.2.3.2 --- Induction and Assay of Natural Killer Cell Activity --- p.60 / Chapter 2.2.3.3 --- Induction and Assay of Lymphokine-activated Killer Cell Activity --- p.61 / Chapter 2.2.3.4 --- Assay of In Vitro Tumour Cell Proliferation --- p.62 / Chapter 2.2.3.5 --- In Vitro Tumour Clonogenicity Assay --- p.63 / Chapter 2.2.3.6 --- Induction of Myeloid Leukemic Cell Differentiation --- p.63 / Chapter 2.2.3.7 --- DNA Fragmentation Analysis --- p.64 / Chapter 2.2.3.8 --- Cell Cycle and DNA Content Evaluation --- p.66 / Chapter 2.2.4 --- Statistical Analysis --- p.66 / Chapter CHAPTER 3: --- "Extraction, Purification and Characterization of GTCs" / Chapter 3.1 --- INTRODUCTION --- p.67 / Chapter 3.2 --- RESULTS --- p.69 / Chapter 3.2.1 --- "Extraction of Catechins from the Chinese Green Tea, Ji Pin Long Jing" --- p.69 / Chapter 3.2.2 --- Analysis of Epicatechin Isomers by HPLC --- p.69 / Chapter 3.2.3 --- Bio-toxicity Determination by Brine Shrimp Bioassay --- p.71 / Chapter 3.2.4 --- Effect of GTCs on the Viability of Murine Splenocytes --- p.72 / Chapter 3.3 --- DISCUSSION --- p.74 / Chapter CHAPTER 4: --- The Immunomodulatory Activities of GTCs / Chapter 4.1 --- INTRODUCTION --- p.75 / Chapter 4.2 --- RESULTS --- p.77 / Chapter 4.2.1 --- Effects of GTCs on the In Vitro Proliferation of Murine Splenocytes --- p.77 / Chapter 4.2.2 --- In Vitro Co-mitogenic Effect of GTCs on Murine Splenocytes --- p.77 / Chapter 4.2.3 --- Stimulation of Lymphocyte Proliferation In Vitro by Intraperitoneal Administration of GTCs --- p.81 / Chapter 4.2.4 --- Effect of In Vivo Treatment of GTCs on In Vitro Mixed Lymphocyte Reaction --- p.81 / Chapter 4.2.5 --- Effect of In Vivo Administration of GTCs on Splenic Lymphocytes Subpopulations --- p.87 / Chapter 4.2.6 --- Effect of In Vivo Administration of GTCs on the Induction of Alloreactive Cytotoxic T Lymphocytes --- p.87 / Chapter 4.2.7 --- In Vivo Induction of Delayed-type Hypersensitivity (DTH) Response to Sheep Red Blood Cells (SRBC) by GTCs --- p.91 / Chapter 4.2.8 --- Primary Humoral Immune Response to SRBC in GTCs- treated Mice --- p.91 / Chapter 4.2.9 --- Effect of GTCs on In Vivo Migration of Macrophages --- p.95 / Chapter 4.2.10 --- Effect of GTCs on the Phagocytic Activity of Thioglycollate- elicited Macrophages In Vitro --- p.95 / Chapter 4.2.11 --- Effect of In Vivo Administration of GTCs on Phagocytic Activity of Thioglycollate-elicited Macrophages In Vitro --- p.98 / Chapter 4.2.12 --- Effect of GTCs on In Vitro Cytostatic Activity of Picolinic Acid-activated Macrophages --- p.98 / Chapter 4.2.13 --- Effect of In Vivo Administration of GTCs on the Cytostatic Activity of Picolinic Acid-activated Macrophages --- p.101 / Chapter 4.3 --- DISCUSSION --- p.103 / Chapter CHAPTER 5: --- THE ANTI-TUMOUR ACTIVITIES OF GTCs / Chapter 5.1 --- introduction --- p.107 / Chapter 5.2 --- RESULTS --- p.109 / Chapter 5.2.1 --- Effects of GTCs on the Growth of Transplantable Tumour Cells In Vivo --- p.109 / Chapter 5.2.2 --- Effect of GTCs on In Vivo Induction of Natural Killer Cells --- p.109 / Chapter 5.2.3 --- Effect of GTCs on In Vitro Induction of Lymphokine- activated Killer Cell Activity --- p.113 / Chapter 5.2.4 --- In Vitro Cytotoxicity of Green Tea Epicatechin Isomers on Murine and Human Tumour Cell Lines --- p.115 / Chapter 5.2.5 --- In Vitro Cytostatic Effect of EGCG on Various Tumour Cell Lines --- p.118 / Chapter 5.2.6 --- Effect of EGCG on the In Vitro Clonogenicity of Myeloid Leukemia Cells --- p.127 / Chapter 5.2.7 --- Induction of Apoptosis of Myeloid Leukemia HL-60 Cells In Vitro --- p.130 / Chapter 5.2.8 --- Effect of EGCG on the Cell Cycle Kinetics of Myeloid Leukemia HL-60 Cells In Vitro --- p.131 / Chapter 5.2.9 --- Effect of EGCG on the Morphological Changes of Myeloid Leukemia Cells --- p.136 / Chapter 5.2.10 --- Effect of EGCG on the Endocytic Activity of Myeloid Leukemia Cells --- p.136 / Chapter 5.3 --- discussion --- p.141 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPEC- TIVES --- p.145 / REFERENCES --- p.152

Resumo: O presente estudo teve como objetivo avaliar o efeito citotóxico dos monômeros isobutil metacrilato (IBMA) e 1,6 - Hexanediol dimetacrilato (1,6 - HDMA), do plastificante di-n-butil ftalato (DBP), e dos produtos de degradação ácido metacrílico (AM) e ácido benzóico (AB) sobre células L929. Esses compostos foram testados em faixas de concentrações liberadas, em um período de 30 dias, por materiais reembasadores rígidos, previamente quantificadas em estudos anteriores. O efeito citotóxico foi verificado por meio dos testes de MTT e 3H-timidina, após as células terem sido expostas às substâncias testadas nas concentrações estabelecidas. A classificação da citotoxicidade foi baseada na viabilidade celular em relação ao controle (células expostas ao meio sem as substâncias testadas). A atividade de síntese de DNA foi inibida por todos os compostos. Os resultados do presente estudo demonstraram que o teste de 3H-timidina foi mais sensível que o teste de MTT e que os compostos avaliados mostraram diferentes níveis de citotoxicidade in vitro. A atividade da desidrogenase mitocondrial diminuiu nas células tratadas com os monômeros, o plastificante e o produto de degradação AM; porém, para o AB, a maioria das concentrações testadas não apresentou efeito citotóxico. / Abstract: The aim of this study was to evaluate the cytotoxic effect of the monomers 1,6 - hexanediol dimethacrylate (1,6 - HDMA) and isobutyl methacrylate (IBMA), the plasticizer di-n-butyl phthalate (DBP), and the degradation by-products methacrylic acid (MA) and benzoic acid (BA) on L929 cells. These compounds were tested in the range of concentrations released in 30 days from hard chairside reline resins that were quantified in previous investigations. Cytotoxic effects were assessed by using MTT and 3H - thymidine assays after the cells had been exposed to the test compounds at the given concentrations. Cytotoxicity was rated based on cell viability relative to controls (cells exposed to medium without test substances). The results presented in this study demonstrated that the 3H-thymidine assay was more sensitive than the MTT assay, and all compounds tested showed varying degrees of cytotoxic effect in vitro. DNA synthesis activity was inhibited by all compounds. Mitochondrial dehydrogenase activity decreased in cells treated with monomers, plasticizer and MA by-product, whereas no cytotoxic effect was observed on contact with BA at the majority of concentrations tested. / Orientador: Ana Lucia Machado / Coorientador: Iracilda Zeppone Carlos / Banca: Ana Cláudia Pavarina / Banca: Cláudia Lovato da Silva / Mestre

No description available.

Bibliography: leaves 287-310 / 311 leaves : / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (M.D.)--University of Adelaide, Dept. of Paediatrics, 1991

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2002. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 4 uppsatser.