Char Tsui Shan. / Thesis submitted in: October 2000. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 92-114). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iv / Acknowledgements --- p.vi / Table of Contents --- p.vii / Contents of Chapter 1: Introduction --- p.viii / Contents of Chapter 2: Materials & Methods --- p.viii / Contents of Chapter 3: Results --- p.x / Lists of Tables --- p.xi / List of Figures --- p.xii / Chapter Chapter 1: --- Introduction --- p.1-24 / Chapter Chapter 2: --- Material & Methods --- p.25 -41 / Chapter Chapter 3: --- Results --- p.42 -76 / Chapter Chapter 4: --- Discussion --- p.77 -89 / Chapter Chapter 5: --- Future Work --- p.90 -91 / References --- p.92 -107 / Chapter Chapter 1: --- Introduction / Chapter 1.1. --- Taxonomy of Enterococci --- p.1 / Chapter 1.2. --- Natural Habitat of Enterococci --- p.2 / Chapter 1.3. --- Characteristics of Enterococci --- p.2 / Chapter 1.4. --- Identification to Species Level of Enterococci --- p.2 / Chapter 1.5. --- Clinical Significance of Enterococci --- p.5 / Chapter 1.6 --- Clinical Management of Enterococcal Infections --- p.8 / Chapter 1.7. --- Antimicrobial Susceptibilities of Enterococci --- p.8 / Chapter 1.8. --- Antimicrobial Resistance of Enterococci --- p.11 / Chapter 1.9. --- Molecular typing of enterococci --- p.16 / Chapter 1.10. --- Epidemiological Studies of Enterococci --- p.19 / Chapter 1.11. --- Objectives --- p.21 / Chapter Chapter 2. --- Materials & Methods / Chapter 2.1. --- Collection of strains --- p.23 / Chapter 2.2. --- Quality control strains --- p.23 / Chapter 2.3. --- Identification of strains to genus level --- p.23 / Chapter 2.4. --- Identification of strains to species level --- p.24 -27 / Chapter 2.4.1. --- By commercially available biochemical method / Chapter 2.4.2. --- By molecular method 226}0ؤ multiplex PCR / Chapter 2.4.2.1. --- Source of primers / Chapter 2.4.2.2. --- Extraction of bacterial DNA / Chapter 2.4.2.3. --- Multiplex PCR / Chapter 2.4.2.4. --- Analysis of PCR products agarose gel electrophoresis / Chapter 2.4.3. --- By conventional biochemical methods --- p.27 / Chapter 2.5. --- Antimicrobial susceptibility testing --- p.27 -30 / Chapter 2.5.1. --- Preparation of antimicrobial agent stock solution / Chapter 2.5.2. --- Preparation of agar medium / Chapter 2.5.3. --- Preparation of inoculum and inoculation of plates / Chapter 2.5.4. --- Incubation and reading of plates / Chapter 2.6. --- Mechanisms of antibiotic resistance --- p.30 -35 / Chapter 2.6.1. --- Amplification of vancomycin resistance genes in enterococci by multiplex PCR / Chapter 2.6.1.1. --- Isolates / Chapter 2.6.1.2. --- Primers / Chapter 2.6.1.3. --- Extraction of bacterial DNA and PCR / Chapter 2.6.2. --- "Detection of gene coding for aminoglycoside-modifying enzyme (AAC6'-APH2"") by PCR" / Chapter 2.6.2.1. --- Isolates / Chapter 2.6.2.2. --- Primers / Chapter 2.6.2.3. --- Extraction of bacterial DNA and PCR / Chapter 2.6.3. --- Detection of β-lactamase in enterococci / Chapter 2.7. --- Molecular typing of enterococci by pulsed-field gel electrophoresis --- p.35 -37 / Chapter 2.7.1. --- Incorporation of chromosomal DNA into agarose plugs / Chapter 2.7.2. --- Digestion of chromosomal DNA in agarose plugs with restriction enzyme SmaI / Chapter 2.7.3. --- Pulsed-field gel electrophoresis / Chapter 2.7.4. --- Cluster analysis / Chapter 2.7.5. --- Data analysis / Chapter 2.8. --- Research plan --- p.37 / Chapter Chapter 3. --- Results / Chapter 3.1. --- Identification of enterococci by API 20 Strep and PCR --- p.39-46 / Chapter 3.2. --- Antimicrobial susceptibilities of enterococci --- p.47 -54 / Chapter 3.3. --- Detection of vancomycin resistance genes in enterococci by PCR --- p.55 -57 / Chapter 3.4. --- "Detection of gene coding for aminoglycoside-modifying enzyme (AAC6'- APH2"") by PCR" --- p.58 -60 / Chapter 3.5. --- Detection of β-lactamase in enterococci --- p.61 / Chapter 3.6. --- Resistance pattern of enterococci --- p.62 -64 / Chapter 3.7. --- Multiresistant enterococci investigated by pulsed-field gel electrophoresis

Wong, Ah Ting. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (p. 125-130). / Abstracts in English and Chinese. / ABSTRACT --- p.V / 摘要 --- p.VIII / TABLE OF CONTENTS --- p.X / ACKNOWLEDGEMENTS --- p.XII / INTRODUCTION --- p.13 / Adaptive Mutation versus Spontaneous Mutation --- p.13 / Fluctuation Test --- p.13 / Adaptive Reversion of lacI - lacZ Fusion Mutant --- p.17 / Putative Models of Adaptive Mutation Mechanisms --- p.18 / Point Mutagenesis --- p.18 / Hypermutation --- p.19 / Gene Amplification --- p.21 / Controversy over the Mechanism of Adaptive Mutation --- p.21 / Induced Mutagenesis under Other Stresses --- p.23 / The General Stress Response --- p.23 / The SOS Response --- p.24 / Reduced Mismatch Repair --- p.24 / Adaptive Mutation in Other Micro-organisms --- p.25 / Mutation Generation under Antibiotic Stress --- p.26 / Fluoroquinolones --- p.26 / Beta-Lactams --- p.27 / Aminoglycosides --- p.27 / Justification and Objectives of this Study --- p.28 / MATERIAL AND METHODS --- p.30 / Bacterial Strains --- p.30 / Culture Media --- p.30 / Antibiotics --- p.30 / Resistance Induction Assay --- p.31 / Rationale of Experimental Design --- p.31 / Agar Selection Method --- p.32 / Broth Selection Method --- p.34 / Isolation of Organisms which Exhibited Reduced Drug Susceptibility and Determination the Minimal Inhibitory Concentration (MIC) --- p.36 / Indole Test --- p.37 / DNA Extraction --- p.37 / Polymerase Chain Reaction (PCR) on the gyrA and rpoB Genes --- p.37 / PCR Product Purification and Nucleotide Sequencing --- p.39 / RESULTS --- p.40 / Solid Agar Selection Approach --- p.41 / Broth Selection Approach --- p.48 / Strain MG1655 --- p.49 / Strain BW25113 --- p.53 / recA Deletion Mutant --- p.54 / mutS Deletion Mutant --- p.56 / DISCUSSION --- p.59 / Development of Resistance Induction Assay --- p.59 / Background Resistance to Gentamicin and Rifampicin --- p.62 / Resistance Induction Effect of Ciprofloxacin --- p.64 / Relative Effects of recA and mutS Deletion --- p.66 / Putative Origins of Antibiotic Resistance Gene Mutations --- p.69 / TABLES AND FIGURES --- p.71 / REFERENCES --- p.125

Choi, Siu Cheong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 132-148). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / Abbreviation --- p.v / List of Figures --- p.viii / List of Tables --- p.xii / Table of Content --- p.xiii / Chapter Chapter 1: --- General Introduction / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- Treatment resistance in cancer --- p.1 / Chapter 1.1.1.1 --- Surgery --- p.2 / Chapter 1.1.1.2 --- Chemotherapy --- p.3 / Chapter 1.1.1.3 --- Radiotherapy --- p.3 / Chapter 1.1.1.4 --- Hormonal therapy --- p.4 / Chapter 1.1.2 --- Hypoxia/reoxygenation and its correlation with treatment resistance --- p.5 / Chapter 1.1.3 --- Aim of the study --- p.6 / Chapter Chapter 2: --- The drug sensitivity in HepG2 cells and A431 cells / Chapter 2.1 --- Introduction --- p.8 / Chapter 2.1.1 --- Treatment of cancer --- p.8 / Chapter 2.1.2 --- Drug resistance --- p.9 / Chapter 2.2 --- Materials and Methods --- p.10 / Chapter 2.2.1 --- Cell culture --- p.10 / Chapter 2.2.2 --- Drugs --- p.10 / Chapter 2.2.3 --- MTT assay --- p.11 / Chapter 2.3 --- Results --- p.12 / Chapter 2.3.1 --- The drugs to which G10HR and G20HR cells were more resistant --- p.12 / Chapter 2.3.2 --- "The drugs of which GP, G10HR and G20HR cells have similar response" --- p.12 / Chapter 2.3.3 --- The drugs to which A10HR and A20HR cells were more resistant --- p.17 / Chapter 2.3.4 --- The drugs to which A10HR and/or A20HR cells were more sensitive --- p.17 / Chapter 2.3.5 --- "The drugs which AP, A10HR and A20HR cells have similar response" --- p.18 / Chapter 2.4 --- Discussion --- p.24 / Chapter 2.4.1 --- Camptothecin and 10-hydroxy camptothecin --- p.27 / Chapter 2.4.2 --- Etoposide --- p.30 / Chapter 2.4.3 --- Hydrogen peroxide --- p.32 / Chapter 2.4.4 --- Interferons --- p.32 / Chapter 2.4.4.1 --- Interferon alpha --- p.33 / Chapter 2.4.4.2 --- Interferon gamma --- p.34 / Chapter 2.4.5 --- Methotrexate --- p.35 / Chapter 2.4.6 --- Vincristine --- p.36 / Chapter Chapter 3: --- The resistance mechanism of doxorubicin in A431 cells / Chapter 3.1 --- Introduction --- p.38 / Chapter 3.1.1 --- Chemotherapeutic resistance --- p.38 / Chapter 3.1.2 --- Tumor hypoxia --- p.39 / Chapter 3.1.3 --- Structure and function of doxorubicin --- p.39 / Chapter 3.1.4 --- Clinical use of doxorubicin --- p.40 / Chapter 3.1.5 --- Mechanisms of doxorubicin resistance --- p.41 / Chapter 3.1.6 --- Structure and function of P-glycoprotein --- p.42 / Chapter 3.1.7 --- Drug resistance contributed by P-glycoprotein and the solution --- p.43 / Chapter 3.1.8 --- Epigenetic modulation of mdr1 --- p.45 / Chapter 3.2 --- Materials and Methods --- p.47 / Chapter 3.2.1 --- Cell culture --- p.47 / Chapter 3.2.2 --- MTT assay --- p.47 / Chapter 3.2.3 --- Reverse transcription polymerase chain reaction (RT-PCR) --- p.47 / Chapter 3.2.4 --- Western blot analysis --- p.48 / Chapter 3.2.5 --- Doxorubicin efflux assay --- p.50 / Chapter 3.2.6 --- Drug sensitivity of A431 cells treated with verapamil --- p.50 / Chapter 3.2.7 --- Treatment with DNA methyltransferase inhibitor --- p.51 / Chapter 3.2.8 --- Drug sensitivity of A431 cells treated with 5-Aza-dC --- p.51 / Chapter 3.2.9 --- Methylation-specific PCR (MSP) --- p.51 / Chapter 3.2.10 --- Bisulfite genomic DNA sequencing --- p.52 / Chapter 3.3 --- Results --- p.54 / Chapter 3.3.1 --- Drug sensitivity of A431 cells to doxorubicin --- p.54 / Chapter 3.3.2 --- Expression profile of mdrl and P-glycoprotein in A431 cells --- p.54 / Chapter 3.3.3 --- Dox efflux-pump activity in A431 cells --- p.57 / Chapter 3.3.4 --- Drug sensitivity of A431 cells in the presence of verapamil --- p.59 / Chapter 3.3.5 --- Expression profile of mdrl in A431 cells in the presence of 5- Aza-dC --- p.59 / Chapter 3.3.6 --- Drug sensitivity of A431 cells in the presence of 5-Aza-dC --- p.62 / Chapter 3.3.7 --- Methylation status of mdrl promoter region --- p.64 / Chapter 3.3.8 --- Bisulfite genomic DNA sequencing of the mdrl promoter --- p.64 / Chapter 3.4 --- Discussion --- p.67 / Chapter Chapter 4: --- The resistance mechanism of cisplatin in HepG2 cells / Chapter 4.1 --- Introduction --- p.70 / Chapter 4.1.1 --- Tumor hypoxia and chemotherapeutic resistance --- p.70 / Chapter 4.1.2 --- Cisplatin and its action mechanism --- p.71 / Chapter 4.1.3 --- Mechanisms of cisplatin resistance --- p.74 / Chapter 4.1.4 --- Mismatch repair genes --- p.79 / Chapter 4.1.5 --- Epigenome and drug resistance in cancer --- p.80 / Chapter 4.2 --- Materials and Methods --- p.84 / Chapter 4.2.1 --- Cell culture --- p.84 / Chapter 4.2.2 --- MTT assay --- p.84 / Chapter 4.2.3 --- Reverse transcription polymerase chain reaction (RT-PCR) --- p.84 / Chapter 4.2.4 --- Oligonucleotide transfection --- p.85 / Chapter 4.2.5 --- Treatment with DNA methyltransferase inhibitor --- p.86 / Chapter 4.2.6 --- Drug sensitivity of HepG2 cells treated with 5-Aza-dC --- p.87 / Chapter 4.2.7 --- Treatment with histone deacetylase inhibitor --- p.87 / Chapter 4.2.8 --- Drug sensitivity of HepG2 cells treated with TSA --- p.87 / Chapter 4.3 --- Results --- p.89 / Chapter 4.3.1 --- Drug sensitivity of HepG2 cells to cisplatin --- p.89 / Chapter 4.3.2 --- Expression profile of the MMR genes in HepG2 cells --- p.89 / Chapter 4.3.3 --- Drug sensitivity of HepG2 cells to cisplatin after the knock- down of PMS2 --- p.91 / Chapter 4.3.4 --- Expression profile of MMR genes in the presence of 5-Aza-dC --- p.95 / Chapter 4.3.5 --- Drug sensitivity of HepG2 cells to cisplatin after the addition of 5-Aza-dC --- p.95 / Chapter 4.3.6 --- Expression profile of MMR genes in the presence of trichostatin A --- p.98 / Chapter 4.3.7 --- Sensitivity of HepG2 cells to cisplatin after the addition of trichostatin A --- p.98 / Chapter 4.4 --- Discussion --- p.101 / Chapter Chapter 5: --- The role of PMS2 in cisplatin-induced apoptosis / Chapter 5.1 --- Introduction --- p.105 / Chapter 5.1.1 --- Apoptosis --- p.105 / Chapter 5.1.2 --- Extrinsic pathway of apoptosis --- p.106 / Chapter 5.1.3 --- Intrinsic pathway of apoptosis --- p.106 / Chapter 5.1.4 --- Cisplatin-induced apoptosis --- p.107 / Chapter 5.1.5 --- MMR and apoptosis --- p.109 / Chapter 5.2 --- Materials and Methods --- p.111 / Chapter 5.2.1 --- Cell culture --- p.111 / Chapter 5.2.2 --- Flow cytometric analysis of apoptosis --- p.111 / Chapter 5.2.3 --- Oligonucleotide transfection --- p.111 / Chapter 5.2.4 --- Western blot analysis --- p.111 / Chapter 5.2.5 --- Drug and antibodies --- p.112 / Chapter 5.3 --- Results --- p.113 / Chapter 5.3.1 --- Cisplatin induced apoptosis --- p.113 / Chapter 5.3.2 --- Knockdown of PMS2 by siRNA --- p.113 / Chapter 5.3.3 --- Cisplatin-induced apoptosis involved caspases --- p.115 / Chapter 5.3.4 --- Protein expressions of anti-apoptotic genes --- p.119 / Chapter 5.3.5 --- Protein expressions of pro-apoptotic genes --- p.119 / Chapter 5.3.6 --- Protein expressions of apoptotic proteins after knockdown of PMS2 --- p.122 / Chapter 5.4 --- Discussion --- p.124 / Chapter Chapter 6: --- General discussion and conclusion / Chapter 6.1 --- Diverse sensitivity for hypoxia/reoxygenation treated cells to anticancer drugs --- p.128 / Chapter 6.2 --- Resistance mechanism of doxorubicin in A10HR and A20HR cells --- p.129 / Chapter 6.3 --- Resistance mechanism of cisplatin in G10HR and G20HR cells --- p.129 / Chapter 6.4 --- The role of PMS2 as a direct signaling molecule and the alteration of apoptotic proteins in cisplatin-induced apoptosis --- p.130 / Chapter 6.5 --- Future work --- p.131 / References --- p.132

Leung, Siu Sing. / Thesis M.Phil. Chinese University of Hong Kong 2015. / Includes bibliographical references (leaves 92-97). / Abstracts also in Chinese. / Title from PDF title page (viewed on 25, October, 2016).

鮑曼不動桿菌(Acinetobacter baumannii)是一種威脅生命的醫院獲得性病菌。該細菌有很强的環境適應能力。它能夠在重症監護室被分離出來并有很高的幾率感染免疫系統受損的病人。鮑曼不動桿菌有很高的傾向獲得多重抗藥性。目前在亞洲和歐洲有多株泛抗藥性菌株被發現。一些基因組比對研究著重報告了鮑曼不動桿菌的抗藥基因片段和與抗藥性相關的基因突變。然而，抗藥基因的轉錄調控和該細菌在抗生素治療過程中引發的反應并未得到很好的研究。因此，我們運用鏈特異性轉錄組測序技術（RNA-seq）對一些抗藥菌株和非抗藥菌株在不同環境下生長的樣本進行測序，來研究該細菌，尤其是在抗生素治療中抗藥菌株的基因轉錄調控。 / 本研究運用轉錄組測序技術（RNA-seq）系統分析了十二株鮑曼不動桿菌在培養液生長狀況下的轉錄組。本次研究共收集了九株多重抗藥性菌株和三株敏感菌株，其中包括了一些快速生長的菌株和慢速生長的菌株。在快速生長的菌株中，氨基酸代謝途徑、甘油脂代謝途徑和钳铁化合物生物合成途徑被向上調控並扮演着重要角色。多重抗藥性菌株擁有更多與轉位酶（transposase）相關的抗生素抗性基因，但除此之外，在對數期的生長過程中多重抗藥性菌株與敏感菌株並未在許多其他代謝途徑中表現差異性控制。 / 三株擁有相同脉冲场凝胶电泳（PFGE）樣式但是表現出不同抗藥性的菌株分別生長於含有阿米卡星(Amikacin)、亞胺培南（Imipenem）或美羅培南（Meropenem）的培養液中，然後它們的轉錄組也被進行了研究。菌株生長在含有抗生素的培養液中時，與能量製造相關的的途徑和核醣體合成途徑被向上調控。作用機制不同的抗生素對細菌有不同的影響，阿米卡星誘發更多基因被向上調控，例如與蛋白質折疊相關的基因；碳青霉烯类抗生素誘發更多的基因被向下調控，例如甘油脂代謝途徑。然而，許多在抗生素治療過程中被緊密調控的基因功能仍舊未知。在抗生素環境生長的條件下基因調控和抗藥機制可能會更複雜。 / 最後，本研究找到一些新的與抗藥性相關的基因和单核苷酸变异（SNVs）。其中，源自於同一操縱子的大环内酯二位轉磷酸酶（macrolide2’ phosphotransferase）同源體Mph和大环内酯外排泵蛋白同源體Mel只存在並一同表達於鮑曼不動桿菌的阿米卡星抗藥株中。這兩個基因或對阿米卡星的抗藥性有一定貢獻作用。總而言之，這些成果爲將來的深度研究提供了重要依據。 / Acinetobacter baumannii is a life-threatening nosocomial pathogen, which has versatile adaptability to the environment. It can be isolated from intensive care unit (ICU) and causes high prevalence of infection among immunocompromised patients. A. baumannii has high tendency to develop multidrug resistance. Currently, pan-drug resistant strains have been reported in Asia and Europe. Several comparative genomic studies revealed the structures of drug resistant islands and antibiotic-related mutations in A. baumannii. However, the transcriptional regulation of drug resistant genes, and the multidrug resistant response of A. baumannii under the treatment of antibiotics are not well studied. By applying strand-specific RNA-sequencing on sensitive and multidrug resistant strains growing in various conditions, we aimed to study the transcriptional responses and gene regulation of A. baumannii, specifically under the antibiotic treatment. / The transcriptome of twelve A. baumannii strains, including nine multidrug resistant strains and three sensitive strains, were systematically analyzed in planktonic state by RNA-seq. Among the multidrug resistant strains there are both fast-and slow-growing strains. Amino acid metabolic pathways, glycerol lipid metabolic pathways and siderophore biosynthetic process are found to be key pathways that are up-regulated in fast-growing strains. Except that multidrug resistant strains possess more transposase-associated antibiotic resistant genes, intriguingly, only a few pathways are differentially regulated between multidrug resistant and sensitive strains during fast growth in antibiotic-free medium. / Three strains of the same PFGE pattern but with different antibiotic resistance patterns were treated by amikacin, imipenem, and meropenem, and their transcriptomes were analyzed. The energy generation-related pathways and ribosome synthesis pathway were commonly up-regulated when the strains were grown in antibiotic-treated media. Amikacin triggers more genes up-regulated, including genes responsible for protein folding, while carbapenems trigger more genes down-regulated, including glycerol lipid metabolic process, revealing the different actions of antibiotics. However, many tightly-regulated genes during antibiotic treatment were functionally unknown, suggesting that gene regulation during antibiotic response and the actual mechanisms involved could be far more complex. / Finally, this study also identified several novel genes and single nucleotide variations (SNVs) which were correlatedto antibiotic-specific resistance. A macrolide 2’ phosphotransferase homolog Mph and a macrolide efflux protein homolog Mel, which commonly exist only in A. baumannii amikacin resistant strains and are co-expressed in the same operon, may contribute to amikacin resistance. In summary, the results presented in this thesis have opened the venue for future investigations. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Qin, Hao. / Thesis (Ph.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 107-114). / Abstracts also in Chinese.

Chan, Lap Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 131-147). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.iii / ABSTRACT (CHINESE VERSION) --- p.v / TABLE OF CONTENTS --- p.vii / LIST OF TABLES --- p.xiv / LIST OF FIGURES --- p.xv / LIST OF ABBREVIATIONS --- p.xvii / Chapter CHAPTER 1 --- INTRODUCTION / Chapter 1.1 --- Background --- p.2 / Chapter 1.2 --- Prevalence of antimicrobial reisitance in gonococcal strains --- p.4 / Chapter 1.2.1 --- Prevalence of penicillin resistant gonococcal strains --- p.5 / Chapter 1.2.2 --- Prevalence of tetracycline resistant gonococcal strains --- p.6 / Chapter 1.2.3 --- Prevalence of quinolone resistant gonococcal strains. --- p.7 / Chapter 1.2.4 --- Emergence of generation cephalosporin reduced susceptible gonococcal strains --- p.8 / Chapter 1.2.5 --- Monitoring the prevalence of gonorrhea --- p.9 / Chapter 1.3 --- Gonococcal Antimicrobial Surveillance Program --- p.10 / Chapter 1.4 --- Antimicrobial Resistance Mechanisms in N. gonorrhoeae --- p.11 / Chapter 1.4.1 --- "Innate Resistance Mechanisms in N, gonorrhoeae" --- p.11 / Chapter 1.4.1.1 --- Natural mechanisms in N. gonorrhoeae against toxic substance --- p.11 / Chapter 1.4.1.2 --- Efflux pump inhibitors --- p.13 / Chapter 1.4.2 --- Development of acquired antimicrobial resistance in N. gonorrhoeae --- p.14 / Chapter 1.4.2.1 --- Penicillin --- p.14 / Chapter 1.4.2.1.1 --- Chromosomal-mediated resistance --- p.14 / Chapter 1.4.2.1.2 --- Plasmid-mediated resistance --- p.16 / Chapter 1.4.2.2 --- Tetracycline --- p.16 / Chapter 1.4.2.2.1 --- Plasmid-mediated resistance --- p.17 / Chapter 1.4.2.2.2 --- Chromosomal mediated resistance --- p.18 / Chapter 1.4.2.3 --- Fluroquinolone --- p.18 / Chapter 1.4.2.3.1 --- Resistant mechanism in quinolone resistant gonococcal strains --- p.19 / Chapter 1.4.2.3.1.1 --- gyrA andparC --- p.19 / Chapter 1.4.2.3.1.2 --- NorM efflux system --- p.21 / Chapter 1.4.2.4 --- 3rd generation cephalosporins --- p.22 / Chapter 1.4.2.4.1 --- Mosaic penA structure in reduced susceptible gonococcal strains --- p.22 / Chapter 1.4.2.4.2 --- Other mechanisms related to reduced susceptibility in gonococcal strains --- p.24 / Chapter 1.5 --- Application of molecular typing methods to study the epidemiology of N. gonorrhoeae --- p.24 / Chapter 1.5.1 --- Opa typing --- p.25 / Chapter 1.5.2 --- K gonorrhoeae Multi-Antigen Sequence Typing --- p.25 / Chapter 1.6 --- Project Objectives --- p.27 / Chapter CHAPTER 2. --- MATERIALS AND METHODS / Chapter 2.1 --- Collecting gonococcal strains --- p.29 / Chapter 2.2 --- Culturing of N. gonorrhoeae --- p.29 / Chapter 2.3 --- Identification --- p.29 / Chapter 2.3.1 --- Gram staining test --- p.30 / Chapter 2.3.2 --- Oxidase test --- p.31 / Chapter 2.3.3 --- Cabohydrate utilization test --- p.31 / Chapter 2.4 --- In-vitro antimicrobial susceptibility testing --- p.32 / Chapter 2.4.1 --- Preparation of cell cultures for MIC tests --- p.32 / Chapter 2.4.2 --- Preparation of antimicrobial agents for MIC tests --- p.32 / Chapter 2.4.3 --- Inoculum preparation and delivering --- p.33 / Chapter 2.5 --- Preparation of genomic DNA for detection of mutations --- p.34 / Chapter 2.6 --- Study of Resistant Mechanism against fluoroquinolone --- p.34 / Chapter 2.6.1 --- PGR detection of mutations in gyrA and parC genes --- p.35 / Chapter 2.6.2 --- Optimization of gyrA and parC genes PGR --- p.35 / Chapter 2.6.3 --- Detection of PGR products for gyrA and parC genes --- p.37 / Chapter 2.6.4 --- Purification of Amplified DNA products --- p.37 / Chapter 2.7 --- Tests of efflux inhibitor on N. gonorrhoeae --- p.38 / Chapter 2.7.1 --- Effect ofCCCP --- p.39 / Chapter 2.7.2 --- Effect of Reserpine --- p.39 / Chapter 2.8 --- Study of Resistant mechanism against β-lactams --- p.40 / Chapter 2.8.1. --- Detection for the presence of β-lactamase --- p.40 / Chapter 2.8.2 --- Mosaic penA patterns --- p.40 / Chapter 2.8.2.1 --- Detection of mutations in penA gene --- p.40 / Chapter 2.8.2.2 --- Optimization of penA gene PGR --- p.41 / Chapter 2.8.2.3 --- Detection of PGR products --- p.43 / Chapter 2.8.2.4 --- Purification of Amplified DNA products --- p.44 / Chapter 2.9 --- Detection of the presence of tetM determinant --- p.45 / Chapter 2.9.1 --- Optimization of tetM determinant PCRs --- p.46 / Chapter 2.9.2 --- Detection of PGR products --- p.47 / Chapter 2.10 --- Detection of different allele types in tbpB andpor genes --- p.48 / Chapter 2.10.1 --- Optimization of PGR for NG-MAST --- p.48 / Chapter 2.10.2 --- Detection of PCR products --- p.49 / Chapter 2.10.3 --- PCR product purification --- p.50 / Chapter 2.11 --- Sequencing of the PCR products --- p.51 / Chapter 2.12 --- Data Analysis --- p.52 / Chapter CHAPTER 3. --- RESULTS / Chapter 3.1 --- Gonococcal strains collected --- p.55 / Chapter 3.2 --- Identification of gonococcal strains --- p.55 / Chapter 3.3 --- MIC of Antimicrobial agents --- p.56 / Chapter 3.3.1 --- Interpretive Criteria --- p.56 / Chapter 3.3.2 --- Ciprofloxacin --- p.56 / Chapter 3.3.3 --- Penicillin --- p.57 / Chapter 3.3.4 --- Tetracycline --- p.57 / Chapter 3.3.5 --- Ceftriaxone --- p.58 / Chapter 3.3.6 --- Cefixime --- p.58 / Chapter 3.3.7 --- Cefotaxime --- p.58 / Chapter 3.3.8 --- Spectinomycin --- p.58 / Chapter 3.3.9 --- Levofloxacin --- p.59 / Chapter 3.3.10 --- Ceftibuten --- p.59 / Chapter 3.4 --- Result of PGR --- p.60 / Chapter 3.4.1 --- gyrA andparC genes --- p.60 / Chapter 3.4.2 --- penA gene --- p.61 / Chapter 3.4.3 --- tbpB and por genes --- p.62 / Chapter 3.4.4 --- tetM determinant --- p.62 / Chapter 3.5 --- β-lactamase --- p.63 / Chapter 3.6 --- Efflux pump inhibitor --- p.63 / Chapter 3.6.1 --- CCCP --- p.64 / Chapter 3.6.2 --- Reserpine --- p.64 / Chapter 3.7 --- Detection of Mutations --- p.66 / Chapter 3.7.1 --- gyrA and parC genes --- p.66 / Chapter 3.7.2 --- penA gene --- p.68 / Chapter 3.8 --- NG-MAST --- p.70 / Chapter 3.8.1 --- tbpB and por gene --- p.71 / Chapter 3.9 --- Porin mutation --- p.72 / Chapter CHAPTER 4. --- DISCUSSION / Chapter 4.1 --- Sampling --- p.74 / Chapter 4.2 --- Methodology --- p.75 / Chapter 4.3 --- MIC distribution of different antimicrobial agents --- p.76 / Chapter 4.4 --- Mechanisms of quinolone resistance --- p.78 / Chapter 4.4.1 --- Mutations at QRDRs --- p.78 / Chapter 4.4.2 --- Association of the number of mutations at parC gene with MIC levels against fluroquinolones --- p.80 / Chapter 4.5 --- Penicillin and Tetracycline Resistant Mechanisms --- p.81 / Chapter 4.6 --- Efflux system --- p.85 / Chapter 4.7 --- NG-MAST --- p.88 / Chapter 4.8 --- Mosaic penA pattern --- p.89 / Chapter 4.9 --- Management of gonorrhea --- p.90 / Chapter CHAPTER 5. --- CONCLUSIONS / REFERENCES

Thesis (M.Sc.)--Memorial University of Newfoundland, 2005. / Includes bibliographical references.

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 3 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.