by To Siu Hang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 84-92). / Abstract also in Chinese. / Declaration --- p.i / Abstract --- p.ii / Acknowledgement --- p.vi / Abbreviations --- p.vii / List of Figures --- p.viii / List of Tables --- p.xii / Content --- p.xiv / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1. --- Osteosarcoma --- p.1 / Chapter 1.1.1. --- Incidence / Chapter 1.1.2. --- Age and Sex Distribution / Chapter 1.1.3. --- Clinical Features / Chapter 1.1.4. --- Treatment / Chapter 1.2. --- Adriamycin --- p.9 / Chapter 1.2.1. --- Drug Action / Chapter 1.2.2. --- Pharmacology / Chapter 1.3. --- Multidrug Resistance --- p.11 / Chapter 1.4. --- P-glycoprotein --- p.13 / Chapter 1.4.1. --- Nature / Chapter 1.4.2. --- Tissue Distribution / Chapter 1.4.3. --- Relation with MDR / Chapter 1.5 --- Multidrug Resistance Protein --- p.16 / Chapter 1.6. --- Reactive Oxygen Species --- p.17 / Chapter 1.6.1. --- Problems Arising from ROS / Chapter 1.6.2. --- Oxidative Stress and Diseases / Chapter 1.6.3. --- Defense System / Chapter 1.6.4. --- Antioxidative Enzymes / Chapter 1.6.5. --- Relation with MDR / Chapter 1.7. --- Topoisomerase II --- p.22 / Chapter 1.8. --- Methods to Detect MDR --- p.24 / Chapter 1.8.1. --- P-glycoprotein Immunohistochemistry / Chapter 1.8.2. --- Adriamycin Binding Assay / Chapter 1.9. --- Aims of Study --- p.25 / Chapter 2. --- MATERIALS AND METHODS --- p.27 / Chapter 2.1. --- Clinical Study --- p.27 / Chapter 2.1.1. --- Patients Recruitment / Chapter 2.1.2. --- Adriamycin Binding Assay / Chapter 2.1.3. --- P-glycoprotein Immunohistochemistry / Chapter 2.1.3.1. --- Sample and Control Preparation / Chapter 2.1.3.2. --- Immunohistochemical Procedure / Chapter 2.1.4. --- Tumour Necrosis Assessment / Chapter 2.2. --- Effect of Adriamycin on Osteosarcoma Cells --- p.32 / Chapter 2.2.1. --- Establishment of Adriamycin Adapted Osteosarcoma Cells / Chapter 2.2.1.1. --- Maintenance and Subculture of SaOS-2 Cell Line / Chapter 2.2.1.2. --- Storage of Cell Line / Chapter 2.2.1.3. --- Adriamycin Treatment / Chapter 2.2.2. --- KB-V1 Cell Culture / Chapter 2.2.3. --- Adriamycin Binding Assay / Chapter 2.2.4. --- P-glycoprotein Immunohistochemistry / Chapter 2.2.4.1. --- Sample and Control Preparation / Chapter 2.2.4.2. --- Immunohistochemical Procedures / Chapter 2.2.5. --- Thymidine Incorporation Assay / Chapter 2.2.5.1. --- Assay Procedures / Chapter 2.2.6. --- Catalase Assay / Chapter 2.2.6.1. --- Assay Procedures / Chapter 2.2.6.2. --- Unit Calculation / Chapter 2.2.7. --- Glutathione Peroxidase Assay / Chapter 2.2.7.1. --- Assay Procedures / Chapter 2.2.7.2. --- Unit Calculation / Chapter 2.2.8. --- Protein Determination / Chapter 2.3. --- Statistical Analysis --- p.45 / Chapter 3. --- RESULTS --- p.46 / Chapter 3.1. --- Clinical Study --- p.46 / Chapter 3.1.1. --- Patients Recruitment / Chapter 3.1.2. --- Correlation of Adriamycin Sensitivity to Tumour Necrosis / Chapter 3.1.3. --- Correlation of P-glycoprotein Expression to Tumour Necrosis / Chapter 3.1.4. --- Correlation of P-glycoprotein Expression to Adriamycin Sensitivity / Chapter 3.2. --- Effect of Adriamycin on Osteosarcoma Cells --- p.63 / Chapter 3.2.1. --- Adriamycin Sensitivity and P-glycoprotein Expression / Chapter 3.2.2. --- Thymidine Incorporation Rate / Chapter 3.2.3. --- Intracellular Concentration of Catalase / Chapter 3.2.4. --- Intracellular Concentration of Glutathione Peroxidase / Chapter 4. --- DISCUSSIONS --- p.71 / Chapter 4.1. --- Clinical Study --- p.71 / Chapter 4.1.1. --- Patients Recruitment / Chapter 4.1.2. --- Correlation between Adriamycin Sensitivity and Tumour Necrosis / Chapter 4.1.3. --- Correlation between P-glycoprotein Expression and Tumour Necrosis / Chapter 4.1.3.1. --- P-glycoprotein Is Induced During Chemotherapy / Chapter 4.1.3.2. --- P-glycoprotein Cannot Serve As a Prognostic Factor / Chapter 4.1.4. --- Correlation Between Adriamycin Sensitivity and P-glycoprotein Expression / Chapter 4.2. --- Effect of Adriamycin on Osteosarcoma Cells --- p.76 / Chapter 4.2.1. --- Adriamycin Sensitivity and P-glycoprotein Expression / Chapter 4.2.2. --- Proliferation Rate / Chapter 4.2.3. --- Antioxidative Enzymes Activities / Chapter 5. --- CONCLUSION --- p.82 / Chapter 6. --- FURTHER STUDY --- p.83 / Chapter 7. --- BIBLIOGRAPHY --- p.84 / Chapter 8. --- APPENDIX - SOLUTIONS PREPARATION --- p.93

by Cheung Yuen-Nei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 194-208). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Contents --- p.vi / List of Figures --- p.xii / List of Tables --- p.xv / Abbreviations --- p.xvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1 --- Introduction --- p.2 / Chapter 1.1 --- Characteristic of Saponins --- p.3 / Chapter 1.1.1 --- Occurrence of Saponins --- p.3 / Chapter 1.1.2 --- General Properties of Saponins --- p.3 / Chapter 1.1.2.1 --- Emulsifying Agents --- p.3 / Chapter 1.2.2.2 --- Forming Complex with Cholesterol --- p.4 / Chapter 1.1.2.3 --- Hemolytic Property --- p.4 / Chapter 1.1.3 --- Structure of Saponins --- p.5 / Chapter 1.1.3.1 --- Categories of Saponins --- p.5 / Chapter 1.1.3.1.1 --- Triterpene Saponins --- p.5 / Chapter 1.1.3.1.2 --- Steroid Saponins --- p.5 / Chapter 1.1.3.2 --- Monodesmosidic and Bidesmosidic Saponins --- p.7 / Chapter 1.1.4 --- Biological and Pharmacological Properties of Saponins --- p.9 / Chapter 1.1.4.1 --- Anti-microbial Activity --- p.9 / Chapter 1.1.4.1.1 --- Anti-fungal Activities --- p.9 / Chapter 1.1.4.1.2 --- Anti-bacterial Activities --- p.10 / Chapter 1.1.4.1.3 --- Anti-viral Activities --- p.10 / Chapter 1.1.4.2 --- Insecticidal Activity --- p.10 / Chapter 1.1.4.3 --- Molluscicidal Activity --- p.10 / Chapter 1.1.4.4 --- Hypocholesterolemic Activity --- p.11 / Chapter 1.1.4.5 --- Anti-ulcer Activity --- p.11 / Chapter 1.1.4.6 --- Contraceptive Activity --- p.12 / Chapter 1.1.4.7 --- Immunomodulatory Activities --- p.12 / Chapter 1.1.4.7.1 --- Direct Immunostimulation --- p.12 / Chapter 1.1.4.7.2 --- Acting as Immuno-adjuvants --- p.13 / Chapter 1.1.4.8 --- Anti-tumor Activity --- p.14 / Chapter 1.1.4.8.1 --- Anti-carcinogenesis --- p.15 / Chapter 1.1.4.8.2 --- Suppression of Tumor Growth --- p.16 / Chapter 1.1.5 --- Anti-tumor Activity of Steroid Saponins --- p.18 / Chapter 1.1.5.1 --- Diosgenin Steroid Saponin --- p.18 / Chapter 1.1.5.2 --- Hong Kong Compounds --- p.18 / Chapter 1.1.5.3 --- Hong Kong18 --- p.21 / Chapter 1.2 --- Human Hepatocellular Carcinoma (HCC) --- p.24 / Chapter 1.2.1 --- The Incidence of Liver Cancer --- p.24 / Chapter 1.2.2 --- Classification of Liver Cancer --- p.24 / Chapter 1.2.3 --- Human Hepatocellular Carcinoma Cell Lines --- p.25 / Chapter 1.2.3.1 --- Human Hepatocellular Carcinoma Cell Line HepG2 --- p.25 / Chapter 1.2.3.2 --- Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.27 / Chapter 1.2.3.2.1 --- Mechanisms of Multidrug Resistance --- p.28 / Chapter 1.2.3.2.2 --- Structure and Characteristics of P-glycoprotein --- p.29 / Chapter 1.2.3.2.3 --- Methods in Dealing with P-glycoprotein Over-expressed MDR Cells --- p.31 / Chapter 1.3 --- Objectives of the Project --- p.32 / Chapter 1.3.1 --- Study of the Anti-tumor Activities of Hong Kong 18 on Human Hepatocellular Carcinoma Cell Line HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter 1.3.2 --- Study of the Anti-tumor Activities of Hong Kong 18on Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter Chapter 2 --- Materials and Methods --- p.33 / Chapter 2.1 --- Materials --- p.34 / Chapter 2.1.1 --- Cell Culture --- p.34 / Chapter 2.1.1.1 --- Cell Lines --- p.34 / Chapter 2.1.1.2 --- Culture Media --- p.35 / Chapter 2.1.2 --- Reagents and Buffers --- p.36 / Chapter 2.1.2.1 --- Phosphate Buffered Saline (PBS) --- p.36 / Chapter 2.1.2.2 --- Reagents and Buffers for DNA Fragmentation --- p.36 / Chapter 2.1.2.3 --- Reagents and Buffers for Western Analysis --- p.37 / Chapter 2.1.2.4 --- Reagents and Buffer for Caspases Activities --- p.39 / Chapter 2.1.2.5 --- Fluorescent Dyes used for Flow Cytometry --- p.39 / Chapter 2.1.3 --- Chemicals --- p.39 / Chapter 2.2 --- Methods --- p.46 / Chapter 2.2.1 --- MTT Assay --- p.46 / Chapter 2.2.2 --- Determination of Cell Viability --- p.46 / Chapter 2.2.3 --- Purification of Macrophages from balb/c Mice --- p.47 / Chapter 2.2.4 --- Hemolysis Assay --- p.47 / Chapter 2.2.5 --- In vivo Studies of the Toxicity of HK18 --- p.48 / Chapter 2.2.6 --- DNA Fragmentation Assay --- p.50 / Chapter 2.2.7 --- Detection of Apoptotic and Necrotic / Late Apoptotic Cells Death by Flow Cytometry with Annexin V-FITC / PI --- p.51 / Chapter 2.2.8 --- Detection of Mitochondrial Membrane Potential by JC-1 Fluorescent Dye --- p.52 / Chapter 2.2.9 --- Detection of Intracellular Ca Level by Flow Cytometry with Fluo-3 Fluorescent Dye --- p.52 / Chapter 2.2.10 --- Detection of Intracellular Hydrogen Peroxide Level by Flow Cytometry with DCF Fluorescence Dye --- p.53 / Chapter 2.2.11 --- Simultaneous Detection of Mitochondrial Membrane Potential and Intracellular Ca2+ or Mitochondrial Membrane Potential and Intracellular Hydrogen Peroxide --- p.54 / Chapter 2.2.12 --- Western Analysis --- p.55 / Chapter 2.2.12.1 --- Total Protein Extraction --- p.55 / Chapter 2.2.12.2 --- Extraction of Cytosolic Proteins --- p.59 / Chapter 2.2.13 --- Determination of Caspases Enzymatic Activity --- p.63 / Chapter 2.2.14 --- Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) --- p.67 / Chapter 2.2.14.1 --- RNA Extraction by TRIzol Reagent --- p.67 / Chapter 2.2.14.2 --- Reverse Transcription --- p.68 / Chapter 2.2.14.3 --- Polymerase Chain Reaction --- p.68 / Chapter 2.3 --- Statistic Analysis --- p.71 / Chapter Chapter 3 --- Cytotoxicity of HK18 --- p.72 / Chapter 3.1 --- Cytotoxicity of HK18 on HepG2 Cells --- p.73 / Chapter 3.1.1 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by MTT Assay --- p.73 / Chapter 3.1.2 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by Tryphan Blue Exclusion Assay --- p.76 / Chapter 3.2 --- Cytotoxicity of HK18 on R-HepG2 Cells --- p.78 / Chapter 3.2.1 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by MTT Assay --- p.78 / Chapter 3.2.2 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by Tryphan Blue Exclusion Assay --- p.81 / Chapter 3.3 --- Cytotoxicity of HK18 on Macrophages --- p.83 / Chapter 3.4 --- Hemolytic Activity of HK18 --- p.85 / Chapter 3.5 --- In vivo Study of the Toxicity of HK18 --- p.87 / Chapter Chapter 4 --- Mechanistic Study of HK18 on HepG2 Cells --- p.90 / Chapter 4.1 --- Hallmarks of Apoptosis Induced by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.2 --- Induction of DNA Fragmentation by HK18 of HepG2 Cells --- p.97 / Chapter 4.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in HepG2 Cells --- p.99 / Chapter 4.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of HepG2 Cells --- p.99 / Chapter 4.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in HepG2 Cells --- p.101 / Chapter 4.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on HepG2 Cells --- p.105 / Chapter 4.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated HepG2 Cells --- p.108 / Chapter 4.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated HepG2 Cells --- p.114 / Chapter 4.2.1.5 --- HK18 Induced Cytochrome c and AIF Released from Mitochondria of HepG2 Cells --- p.120 / Chapter 4.3 --- Downstream Biochemical Changes Induced by HK18 on HepG2 Cells --- p.123 / Chapter 4.3.1 --- Activation of Caspase 3 of HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.123 / Chapter 4.3.2 --- Induction of Caspases Activities of HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.125 / Chapter 4.4 --- Down-regulation of Anti-apoptotic Bcl-2 Family Members of HepG2 Cells by HK18 --- p.129 / Chapter Chapter 5 --- Mechanistic Study of HK18 on R-HepG2 Cells --- p.133 / Chapter 5.1 --- Hallmarks of Apoptosis Induced by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.2 --- Induction of DNA Fragmentation by HK18 of R-HepG2 Cells --- p.137 / Chapter 5.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in R-HepG2 Cells --- p.139 / Chapter 5.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of R-HepG2 Cells --- p.139 / Chapter 5.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in R-HepG2 Cells --- p.139 / Chapter 5.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on R-HepG2 Cells --- p.142 / Chapter 5.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated R-HepG2 Cells --- p.144 / Chapter 5.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated R-HepG2 Cells --- p.146 / Chapter 5.3 --- Downstream Biochemical Changes Induced by HK18 on R-HepG2 Cells --- p.148 / Chapter 5.3.1 --- Activation of Caspase 3 of R-HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.148 / Chapter 5.3.2 --- Induction of Caspases Activation of R-HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.150 / Chapter 5.4 --- Down-regulation of the Anti-apoptotic Bcl-2 Protein of R-HepG2 Cells by HK18 --- p.154 / Chapter 5.5 --- HK18 was Not a Substrate of P-glycoprotein Contents --- p.156 / Chapter Chapter 6 --- Discussion --- p.158 / Chapter 6.1 --- Cytotoxicity of HK18 --- p.159 / Chapter 6.1.1 --- HK18 was Cytotoxic to the Human Hepatocellular Carcinoma Cell Line HepG2 and Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.159 / Chapter 6.1.2 --- Study of the Toxicity of HK18 --- p.160 / Chapter 6.2 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.1 --- Apoptotic Cell Death Induction of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.162 / Chapter 6.3 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.1 --- Apoptotic Cell Death Induction of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.181 / Chapter Chapter 7 --- Future Perspectives --- p.190 / Chapter Chapter 8 --- References --- p.193

Li Jun. / "March 2002." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (p. 189-216). / 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.

Introdução: Infecções por Enterobactérias resistentes aos carbapenêmicos (ERC), em especial produtoras de Klebsiella pneumoniae carbapenamase tipo KPC hoje são endêmicas em diversas regiões do mundo, seu tratamento é ainda um grande desafio em particular de isolados resistentes à polimixina. Objetivos: Descrever as características clínicas, microbiológicas e moleculares das infecções por ERC. Método: Estudo de coorte prospectiva, realizado no Hospital Universitário de Londrina, Paraná, Brasil, entre março de 2011 a dezembro de 2012. Foram acompanhados pacientes >= 18 anos, que apresentaram infecção por ERC. Dados demográficos e clínicos como idade, sexo, diagnóstico à admissão e presença de co-morbidades de acordo com critérios de Charlson, internação em Unidade de Terapia intensiva e scores APACHE e SOFA desses pacientes, colonização prévia por ERC, cirurgia prévia à infecção, diálise, uso prévio de antimicrobianos e sítio de infecção foram coletados. Foram avaliados os antimicrobianos utilizados para tratamento das infecções por mais de 48 horas nos seguintes pontos: monoterapia ou terapia associada, tempo de início (menor e maior que 12 horas). A identificação do agente foi realizada por método automatizado (Vitek II - bioMerieuxR) e a concentração inibitória mínima dos antibióticos por técnica de microdiluição em caldo, pesquisa de gene blaKPC pela técnica de Polimerase Chain Reaction e sinergismo entre drogas utilizadas em tratamento combinado por meio do método Time Kill. A clonalidade, por Pulsed Field gel eletroforese e analisada por dendograma pelo Bionumerics. Foram realizadas análise bivariada e regressão logística multivariada com técnica de Forward Stepwise para detectar fatores de risco para resistência a polimixina e mortalidade. O nível de significância adotado foi de 5%, utilizando os programas Epi Info 7.0 e SPSS. Resultados: No período de estudo, 127 pacientes apresentaram infecções por ERC, idade média de 55,7 (± 18) anos e 88 (69.3%) do sexo masculino. Infecções de trato respiratório (52-42%) e trato urinário (51 - 40,2%) foram as mais freqüentes, 27 (21,3%) resistentes à polimixina, 113 (89%) das enterobactérias eram K. pneumoniae e 96 (75,6%) tinham gene blaKPC.. Cinquenta e cinco (43,3%) eram polimicrobianas, a maioria (28,3%) co-infecção por Acinetobacter baumannii. A taxa de mortalidade hospitalar foi 61,4%, sendo 34,6% relacionada à infecção e não houve diferença significativa entre os grupos sensíveis (34%) e resistentes à polimixina (37%), p=0.46. Os fatores de risco independentes para óbito foram choque (OR 27.40; IC95% 1.68-446.82; p= 0.02) e diálise (OR 13.26; IC95% 1.17-149.98; p= 0.03); para resistência à polimixina: uso prévio de carbapenem ( OR 2.95; IC95% 1.12-7.78; p= 0.02) e para óbito nessa população: diálise (OR 7,58; IC95% 1,30-43,92; p= 0.02). Terapia combinada, tempo de início de antibiótico sensível e sinergismo in vitro não tiveram impacto significativo na mortalidade. Conclusão: O uso prévio de carbapenêmico foi o único fator associado com a resistência à polimixina nesse estudo. Os fatores associados ao óbito entre os pacientes com infecções por enterobactérias resistentes à polimixina foram fatores de gravidade, como diálise e choque. Nenhuma opção terapêutica, em especial a associação de drogas e nem o tempo de início do tratamento, interferiu na mortalidade deste grupo de pacientes / Introduction: Infections due to Carbapenem resistant Enterobacteriaceae (CRE), particularly Klebsiella pneumoniae producing carbapenemase type KPC, have been endemic in several regions around the world. Their treatment remains a major challenge, particularly for isolates resistant to polymyxin. Objectives: To describe the clinical, microbiological and molecular characteristics of infections by CRE. Methods: Prospective cohort conducted at the University Hospital of Londrina, Paraná, Brazil, from March 2011 to December 2012. All hospitalized patients >= 18 years old who developed infection by CRE were followed until death or discharge. We collected and analyzed the following clinical data: age, sex, diagnosis at admission, presence of comorbidities according to the Charlson criteria, admission in Intensive Care Unit, APACHE and SOFA scores, previous colonization by CRE, previous surgery, dialysis, prior antibiotic use and infection site; furthermore, we also evaluated the time between the blood culture collect and the first antimicrobial dose administration (start time - smaller or longer than 12 hours) as well as whether the treatment was monotherapy or combine therapy for more than 48 hours. The microbiological identification was performed by automated method (Vitek II - bioMerieuxR) and the minimum inhibitory concentration of antibiotics by broth microdilution technique, research blaKPC gene by the technique of Polymerase Chain Reaction and synergism between the drugs used in the combination therapy by Time Kill method. The clonality was carried out by pulsed-field gel electrophoresis and analyzed by dendrogram by BioNumerics. Bivariate analyses and multivariate logistic regression with forward stepwise technique were performed to detect risk factors for resistance to polymyxin and mortality. The level of significance was 5%, using Epi Info 7.0 and SPSS programs. Results: During the study period, 127 patients developed infections by CRE, mean age 55.7 (± 18) years and 88 (69.3%) were male. Respiratory tract infections 52 (42%) and urinary tract 51 (40.2%) were the most frequent. Twenty seven (21.3%) agents were resistant to polymyxin; 113 (89%) were K. pneumoniae and 96 (75.6%) had blaKPC gene. Fifty-five (43.3%) were polymicrobial, the majority (28.3%) co-infection by Acinetobacter baumannii. The hospital mortality rate was 61.4% and 34.6% of the death were related to infection. There was no difference in mortality rate between sensitive (34%) versus resistant (37%)(p = 0.46) to polymyxin. The independent risk factors for death were shock (OR 27.40; 95% CI 1.68-446.82; p = 0.02) and dialysis (OR 13:26; 95% CI 1.17-149.98; p = 0.03); and for resistance to polymyxin were previous use of carbapenem (OR 2.95; 95% CI 1.12-7.78; p = 0.02). The risk factor for death in our study was dialysis (OR 7.58; 95% CI 1.30 to 43.92; p = 0.02). Combine therapy, start time and sensitive and antibiotic synergy in vitro had no significant impact on mortality. Conclusion: In our study, previous carbapenem use was the only factor associated with resistance to polymyxin. Furthermore, dialysis and shock were the only factors associated with death among patients with infections caused by CRE resistant to polymyxin. No therapeutic option, especially the combination of drugs and the start time decreased the higher mortality rates in this group of patients

This thesis summarises research performed with the intent of exploring the many facets of cell death. In the first part of the thesis, the fate of the formin-homology domain containing protein FHOD1 during apoptosis is examined (research performed in the laboratory of Dr. Sophie Roy) and evidence for the cleavage of FHOD1 by caspase-3 at the SVPD616 site is demonstrated. Moreover, the C-terminal FHOD1 cleavage product is shown to translocate to the nucleolus where it inactivates RNA polymerase I transcription. / In the second part of the thesis, the role of the RNA-binding protein HuR in cancer cell migration and invasion, as well as in multidrug resistance is determined using RNA interference to knockdown the expression of HuR in HeLa and KB-V1 cells respectively (research performed in the laboratory of Dr. Imed Gallouzi). In this part of the thesis, HuR is shown to promote cancer cell migration and invasion by stabilizing the beta-actin mRNA in a U-rich-dependent manner. Moreover, evidence is shown for the potential involvement of HuR in the phenomenon of multidrug resistance possibly through the post-transcriptional regulation of the multidrug resistance 1 gene.

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

Thesis (Ph. D.)--Cornell University, May, 1997. / Includes bibliographical references (leaves 170-181).

Thesis (Ph.D. in Toxicology) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 131-147).

Thesis (M.D. with Distinction in Research) -- University of Texas Southwestern Medical Center at Dallas, 2006. / Not embargoed. Vita. Bibliography: 52-62.

No description available.