by Tracy Man-kiu Ma. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 48-61). / Abstract and appendix in English and Chinese. / ABSTRACT --- p.ii / ACKNOWLEDGEMENTS --- p.iii / TABLE OF CONTENTS --- p.iv / LIST OF TABLES --- p.vi / LIST OF FIGURES --- p.vii / LIST OF APPENDICES --- p.viii / Chapter CHAPTER ONE- --- INTRODUCTION / Mesial temporal lobe and its sequel of damages --- p.1 / Mesial temporal lobe pathologies --- p.2 / Memory assessment instruments and the Hong Kong List Learning Test --- p.4 / The receiver operating characteristics (ROC) analysis --- p.6 / Purpose of the present study --- p.7 / Chapter CHAPTER TWO- --- METHOD / Participants --- p.9 / Materials --- p.10 / Procedure --- p.13 / Statistical analysis --- p.15 / Chapter CHAPTER THREE - --- RESULTS / Memory Profiles of NPC Patients with bilateral temporal lobe lesions --- p.18 / Receiver operating characteristics (ROC) analysis for test performance --- p.26 / Validity and reliability --- p.33 / Chapter CHAPTER FOUR - --- DISCUSSION / The clinical utility of the blocked condition --- p.39 / Optimal cutoff scores for sensitivity and specificity --- p.40 / Memory profiles of NPC patients and its implications --- p.42 / Limitations --- p.45 / Conclusions --- p.46 / REFERENCES --- p.48 / APPENDICES --- p.62

Jia, Lin. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 124-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Liu Yuet Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 165-191). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.iii / Abbreviations --- p.vi / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Incidences and Prevalence --- p.2 / Chapter 1.2 --- Anatomy of Uterus and its Support Mechanism --- p.3 / Chapter 1.3 --- Pathophysiology of Uterine Prolapse --- p.5 / Chapter 1.4 --- Classification of Uterine Prolapse --- p.6 / Chapter 1.5 --- Etiology of Uterine Prolapse --- p.7 / Chapter 1.6 --- Treatment of Uterine Prolapse --- p.12 / Chapter 1.6.1 --- Conservative Treatment --- p.12 / Chapter 1.6.2 --- Surgical Treatment --- p.13 / Chapter 1.7 --- Molecular Basis of Uterine Prolapse --- p.14 / Chapter 1.7.1 --- Collagen Metabolism --- p.15 / Chapter 1.7.2 --- Extracellular Matrix Metabolism --- p.16 / Chapter 1.7.3 --- Advanced Glycation End-products --- p.18 / Chapter 1.7.4 --- Estrogen and Estrogen Receptors --- p.19 / Chapter 1.8 --- Gene Expression Profiling of Uterine Prolapse --- p.22 / Chapter 1.9 --- Microarray Gene Expression Profiling Analysis --- p.24 / Chapter 1.9.1 --- Types of Microarray --- p.26 / Chapter 1.9.2 --- Comparison of Oligonucleotide and cDNA Arrays --- p.31 / Chapter 1.10 --- Quantitative Real-time PCR --- p.32 / Chapter 1.10.1 --- Principle of TaqMan Real-time PCR --- p.32 / Chapter 1.10.2 --- Other Types of Real-time PCR --- p.33 / Chapter 1.11 --- Project Aims --- p.34 / Chapter 1.12 --- Significance of Study --- p.35 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.37 / Chapter 2.1 --- Materials --- p.37 / Chapter 2.1.1 --- Patients --- p.37 / Chapter 2.1.2 --- Cardinal Ligament Specimen --- p.38 / Chapter 2.2 --- Methods --- p.39 / Chapter 2.2.1 --- Homogenization of Cardinal Ligament Tissues --- p.39 / Chapter 2.2.2 --- Total RNA extraction --- p.39 / Chapter 2.2.3 --- Oligonucleotide Microarray --- p.41 / Chapter 2.2.3.1 --- Two-cycle cDNA Synthesis --- p.41 / Chapter 2.2.3.2 --- Cleanup of Double-stranded cDNA --- p.45 / Chapter 2.2.3.3 --- Synthesis of Biotin-labeled cRNA --- p.45 / Chapter 2.2.3.4 --- Cleanup and Quantification of Biotin-labeled cRNA --- p.46 / Chapter 2.2.3.5 --- Fragmenting the cRNA for Target Preparation --- p.47 / Chapter 2.2.3.6 --- Target Hybridization --- p.47 / Chapter 2.2.3.7 --- "Array Washing, Staining and Scanning" --- p.48 / Chapter 2.2.3.8 --- Statistical Analysis of Microarray Data --- p.49 / Chapter 2.2.4 --- Quantitative Real-time Polymerase Chain Reaction --- p.52 / Chapter 2.2.4.1 --- Primers and Probes --- p.52 / Chapter 2.2.4.2 --- Reverse Transcription --- p.53 / Chapter 2.2.4.3 --- Plate Setup --- p.53 / Chapter 2.2.4.4 --- Real-time PCR Reaction Mixture Setup --- p.54 / Chapter 2.2.4.5 --- Statistical Analysis of Real-time PCR Data --- p.54 / Chapter CHAPTER 3 --- RESULTS --- p.56 / Chapter 3.1 --- Microarray Gene Expression Data Analysis --- p.57 / Chapter 3.1.1 --- Unsupervised Gene Selection --- p.57 / Chapter 3.1.2 --- Supervised Gene Selection --- p.59 / Chapter 3.1.2.1 --- Gene Expression Profiles Distinguish Cardinal Ligament with Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.59 / Chapter 3.1.2.2 --- Gene Expression Profiles Distinguish Cardinal Ligament with Different Degrees of Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.72 / Chapter 3.1.2.3 --- Gene Expression Profiles Distinguish Cardinal Ligament with Third-degree Prolapse from First-degree Prolapse and Identify Differentially Expressed Genes --- p.92 / Chapter 3.2 --- Validation of Microarray Data by Quantitative Real-time PCR --- p.96 / Chapter 3.2.1 --- Fold Change of Candidate Genes --- p.97 / Chapter 3.2.2 --- Correlation Between Microarray and Quantitative Real-time PCR Results --- p.102 / Chapter CHAPTER 4 --- DISCUSSIONS --- p.103 / Chapter 4.1 --- Global Gene Expression Profiling using Oligonucleotide Microarray --- p.103 / Chapter 4.1.1 --- Advantages of using Affymetrix GeneChipR Microarray for Gene Expression Profiling --- p.103 / Chapter 4.1.2 --- Microarray analysis software --- p.105 / Chapter 4.1.2.1 --- DNA-Chip Analyzer Software --- p.105 / Chapter 4.1.2.2 --- Comparison of Statistical Methods for Analysis of A ffymetrix GeneChipRMicroarray Data --- p.108 / Chapter 4.2 --- Validation of Microarray Data --- p.111 / Chapter 4.2.1 --- Advantages of using Quantitative Real-time PCR for mRNA Quantification --- p.111 / Chapter 4.3 --- Microarray Gene Expression Data Analysis --- p.115 / Chapter 4.3.1 --- Unsupervised Gene Selection --- p.115 / Chapter 4.3.2 --- Supervised Gene Selection --- p.115 / Chapter 4.3.2.1 --- Gene Expression Profiles Distinguish Cardinal Ligament with Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.115 / Chapter 4.3.2.2 --- Gene Expression Profiles Distinguish Cardinal Ligament with Different Degrees of Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.118 / Chapter 4.3.2.3 --- Gene Expression Profiles Distinguish Cardinal Ligament with Third-degree Prolapse from First-degree Prolapse and Identify Differentially Expressed Genes --- p.120 / Chapter 4.4 --- Potential Genes for Further Studies in Uterine Prolapse --- p.120 / Chapter 4.5 --- Implications of This Study --- p.157 / Chapter 4.6 --- Limitations of This Study --- p.160 / Chapter CHAPTER 5 --- CONCLUSIONS --- p.162 / Chapter CHAPTER 6 --- FUTURE PROSPECT --- p.164 / REFERENCES --- p.165

Objetivo: A glicogênio sintase quinase-3 beta (GSK-3B) é uma enzima presente em diversos sistemas biológicos e está envolvida na fisiopatologia de vários transtornos neuropsiquiátricos, incluindo o transtorno bipolar. No entanto, estudos in vivo da GSK-3B que envolvam pacientes bipolares são escassos. O objetivo do presente estudo foi avaliar a atividade da GSK-3B em plaquetas de pacientes idosos com transtorno bipolar em tratamento, em comparação com idosos saudáveis não medicados. Métodos: Foram obtidas amostras de plaquetas de 63 idosos (transtorno bipolar=31, grupo controle=32). A atividade enzimática foi estimada pela razão entre a expressão da forma fosforilada (inativa) da GSK-3B em relação à expressão de ambas as formas (ativa e inativa) da enzima (GSK-3B total), que fornece uma estimativa inversa da atividade enzimática (um aumento da razão indica menor atividade da GSK-3B). A intensidade dos sintomas foi avaliada pela Escala de Depressão de Hamilton de 21 itens e pela Escala de Mania de Young, e o desempenho cognitivo foi avaliado pelo Cambridge Cognitive Test e pelo Mini- Exame do Estado Mental. Resultados: A forma fosforilada da GSK-3B (fosfo-GSK-3B) e a razão da GSK-3B estavam elevadas em pacientes com transtorno bipolar, quando comparadas aos idosos do grupo controle (p=0,018 e p=0,016, respectivamente). Na avaliação por subgrupos, observaram-se níveis da fosfo-GSK-3B e da razão da GSK-3B mais elevados nos pacientes com transtorno bipolar em uso de lítio, quando comparados aos controles (p=0,030 e p=0,023, respectivamente), mas não quando comparados aos pacientes com transtorno bipolar que não usavam lítio. O uso das demais medicações avaliadas (anticonvulsivantes, antipsicóticos, antidepressivos e 16 benzodiazepínicos) não estava associado a diferenças na fosfo-GSK-3B ou na razão da GSK-3B, quando comparado aos controles. Conclusões: A atividade da GSK-3B está diminuída no presente grupo de idosos com transtorno bipolar em tratamento medicamentoso. A ausência de um grupo de pacientes com transtorno bipolar não medicado, e a não uniformidade das medicações utilizadas não nos permitem afirmar se essa redução se deve à características da doença bipolar em si ou seria influência dos medicamentos utilizados / Objective: Glycogen synthase kinase-3 beta (GSK-3B) is an important enzyme present in various biological systems and it is involved in the pathophysiology of many prevalent neuropsychiatric diseases, including bipolar disorder. However, human studies addressing GSK-3B activity in vivo are scarce. The aim of the present study was to evaluate GSK-3B activity in platelets of elderly patients with bipolar disorder undergoing clinical treatment as compared to healthy older adults unmedicated. Methods: Platelets samples where obtained from 63 older adults (bipolar disorder=31, comparison group=32). Enzymatic activity was estimated by means of the ratio between the expression of the phosphorylated (inactive) form of GSK-3B to the expression of both forms (active and inactive) of the enzyme (total GSK-3B), yielding an inverse estimate of enzymatic activity (higher ratio indicating lower GSK- 3B activity). The magnitude of mood symptoms was evaluated by the Hamilton Depression Scale and Young Mania Rating Scale, and the cognitive performance was assessed by the Cambridge Cognitive Test and the Mini-Mental State Examination. Results: The phosphorylated form of GSK-3B (phospho-GSK-3B) and the GSK-3B ratio were elevated in patients with bipolar disorder as compared to healthy controls (P=.018 and P=.016, respectively). When analyzed by subgroups, phospho-GSK-3B and the GSK-3B ratio were elevated in bipolar patients undergoing lithium treatment as compared to healthy controls (P=.030 and P=.023, respectively), but not when compared to bipolar patients without lithium treatment. The use of other drugs evaluated (anticonvulsants, antipsychotics, antidepressants and benzodiazepines) was not associated with distinct values of either phospho-GSK-3B or GSK-3B ratio, when compared to controls. 18 Conclusions: GSK-3B activity is decreased in this group of older adults with bipolar disorder undergoing pharmacological treatment. The absence of a group of unmedicated bipolar patients and the non-uniform pattern of treatment do not allow us to say whether this reduction is due to characteristics of bipolar illness itself or an influence of the therapeutic drugs in use

Further studies were conducted to define an indispensable role of uterine bicarbonate secretion at pre-implantation for the success of blastocyst implantation. The in vitro implantation experiments showed that only when cultured in bicarbonate-containing medium, the blastocysts exhibited normal rate of attachment and outgrowth level. The forskolin-induced endometrial bicarbonate secretion measured by the Isc on pregnant day 4 was almost abolished by CA inhibitor acetazolamide. The efflux of intracellular bicarbonate, measured by intracellular pH-sensitive dye, was blocked by CFTR inhibitor, NPPB, and SLC26a6 inhibitor, DIDS, indicating their involvement in mediating uterine bicarbonate secretion. / In conclusion, the present findings have demonstrated an important role of CFTR in formation of optimal uterine fluid, in terms of both volume and composition, which is crucial for various reproductive events occurring in the uterus. Deviation from the normal uterine fluid composition and volume due to defects in CFTR function or abnormal regulation under pathological conditions, such as CF and genital bacteria infection, probably leads to infertility. The information obtained may provide insight into regulatory mechanism underlying fertility and infertility, as well as the rationale for development of treatment methods for female infertility and new strategies for female contraception. (Abstract shortened by UMI.) / The last part of the study was to demonstrate possible cause of infertility by disturbance of uterine fluid dynamic due to abnormal expression of CFTR using a model of uterine Chlamydia (C.) trachomatis infection, the most common infection-related sterility with the underlying cause unexplained. Uterine C. trachomatis infection induced up-regulated expression of CFTR with enhanced electrolyte and fluid transport as demonstrated by the increase in the cAMP-dependent Isc and uterine wet weight with obvious fluid accumulation in the lumen at diestrus stage, during which the endometrium normally undergoes a series of changes preparing for blastocyst implantation with minimum CFTR expression and uterine fluid volume. The abnormal uterine fluid accumulation upon uterine C. trachomatis infection significantly reduced implantation rate in uterine C. trachomatis infection mouse model. / The present study was aimed to elucidate the cellular and molecular mechanisms underlying the CFTR-related reproductive events in physiological and pathological conditions by using a variety of techniques, including RT-PCR, Western blot, intracellular and extracellular pH measurements, and the short-circuit current (Isc) measurement, in conjunction with mouse primary culture of endometrial cells and blastocyst, as well as several animal models including CF mouse, mouse uterine infectious model and overyectomized (OVX) mouse, etc. / We first examined dynamic changes in uterine bicarbonate secretion, as indicated by bicarbonate-dependent forskolin-induced Isc and epithelial surface pH measurement, and the expression profile of candidate genes and proteins known to be involved in bicarbonate secretion throughout the estrous cycle in mouse uterus. The results showed that the maximum mRNA and protein levels of CFTR, SLC26a6, carbonic anhydrase (CA)2 and CA12 were observed at proestrus stage and/or estrus stages. Luminal surface pH measured by 5-N-hexadecanoyl-aminofluorescein (HAF) showed that the basal endometrial epithelial surface pH at estrus stage was significantly higher than that in diestrus, which could be reduced significantly by CFTR inhibitor DPC, SLC26a6 inhibitor 4',4'-Diisothiocyanostilbene-2',2' Disulfonic Acid (DIDS) and CA suppressor acetazolamide. In the ovariectimized (OVX) mice and primary culture of endometrial cells, estrogen could induce up-regulation of CFTR, SLC26a6, CA2 and CA12 expression with corresponding increase in the bicarbonate-dependent Isc, suggesting a novel role of estrogen in regulating uterine bicarbonate secretion. / He, Qiong. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3247. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 163-176). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Hyperuricemia is the hallmark of gout. Pathogenic mechanisms of hyperuricemia include uric acid overproduction in the liver or underexcretion in the kidney. Current antihyperuricemic agents include xanthine oxidase inhibitors in which allopurinol is the most often prescribed. Inhibitors of renal urate reabsorption such as probenecid and benzbromarone are also employed. However, these existing antihyperuricemic agents possess some undesirable effects such as hypersensitivity towards allopurinol and hepatotoxicity associated with benzbromarone. Therefore, search for alternative antihyperuricemic agents with a more favorable toxicological profile or via mechanisms other than the above two mentioned is highly warranted. / The present project represents such an effort. Four in vitro experimental models were developed for the screening of new antihyperuricemic agents. The effects of the potential compounds from natural sources on the activities of phosphoribosyl pyrophosphate synthetase, hypoxanthine-guanine phosphoribosyl transferase and xanthine oxidase, as well as the uptake of urate through rat renal brush border membrane vesicles were investigated. Several compounds emerged with strong urate uptake inhibitory activities in which morin (3, 5, 7, 2', 4'-pentahydroxyflavone) was the most potent. Interestingly some of these compounds including morin were also demonstrated to be xanthine oxidase inhibitors. The subsequent in vivo experiment showed that morin indeed exhibited hypouricemic and uricosuric actions in an acute oxonate-induced hyperuricemic rat model. The uricosuric action of morin was hirther studied in transfected HEK293 cells expressing the human urate anion transporter 1 (hURATI) which is believed to regulate blood urate level by mediating urate reabsorption. In hURAT1-expressing HEK293 cells, urate uptake was significantly increased as compared to the non-transfected parental cells. Incorporation of morin into the uptake buffer could dose-dependently inhibit urate uptake in the transfected cells. Taken together our data indicated that morin is a potentially useful antihyperuricemic agent which acts by inhibiting xanthine oxidase and inhibiting urate reabsorption. In addition, the favorable safety profile of this natural compound makes it a potential candidate worthy of further investigations. / Yu Zhifeng. / "June 2006." / Advisers: Christopher H. K. Cheng; Wing Ping Fong. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1584. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 155-169). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Frontal lobe atrophy (FLA) is associated with late-life depression and cognitive impairment, although the pathogenesis of FLA in stroke is unclear. In an aim to ascertain whether FLA is affected by WMLs, we analyzed the MRIs of 471 Chinese ischemic stroke patients. Lobar atrophy was defined by a widely-used visual rating scale. WML severity was rated using the Fazekas scale. There was no correlation between PVH and DWMH and temporal and parietal atrophy. The results of this study suggest that FLA in ischemic stroke may be associated with SVD. / Poststroke depression (PSD) is the most common form of poststroke psychiatric morbidity. Small subcortical infarcts (SSIs) can result from small vessel disease (SVD) and large artery disease (LAD). No study has yet explored PSD in different etiological types of SSIs. To address this gap, 127 patients with SSIs resulting from LAD or SVD were examined. PSD was evaluated with the Geriatric Depression Scale (GDS) three months after stroke. The LAD group had a significantly higher frequency of PSD, and LAD was found to be a significant independent risk factor for PSD. This study suggests that cerebral blood perfusion may play an important role in PSD. / Post-stroke emotional lability (PSEL) is a distressing and embarrassing complaint among stroke survivors. Lesions located in various cortical and subcortical areas are thought to be involved in the pathophysiology of PSEL.The clinical significance of microbleeds (MBs) in the development of psychiatric conditions following stroke is unknown. We carried out a study to examine the association between PSEL and MBs in 519 Chinese patients with acute ischemic stroke admitted consecutively. PSEL was evaluated three months after the index stroke, and the number and location of MBs were evaluated with MRI. According to Kim's criteria, 74 (14.3%) of the patients had PSEL. Our results suggest that MBs in the thalamus may play a role in the development of PSEL. The importance of MBs in PSEL and other psychiatric conditions in stroke survivors warrants further investigation. / The first study reported in this thesis involved 328 Chinese ischemic stroke patients who were administered a series of neuropsychological tests covering seven domains three months after stroke. Two hundred and fifty-six of these patients were followed-up for one year. Volumetry of the infarcts, WMLs, and hippocampus atrophy on magnetic resonance imaging (MRI) was conducted. The prevalence of cognitive impairment was 54.9% at baseline and 52.4% at the one-year follow-up, although most of the patients (85.5%) remained cognitively stable. The evolution of cognitive impairment no dementia (CIND) at the one-year follow-up was bidirectional, with 11.2% progressing to dementia and 21.0% reverting to cognitive intact. WMLs volume rather than hippocampal volume was a significant predictor of cognitive impairment, cognitive decline, and delayed dementia. WMLs also had an independent effect on executive function, attention, visual memory, visuoconstruction, and visuomotor speed. / This thesis investigates the clinical and imaging characterization of cognitive impairment and psychiatric morbidity in Chinese stroke patients. The conclusions of the studies reported herein can be summarized as follows. (1) The prevalence of cognitive impairment is high among Chinese poststroke patients, but most remain cognitively stable at one year after stroke; WMLs rather than hippocampal atrophy predict cognitive impairment, longitudinal cognitive decline, and delayed dementia; (2) DLPFC atrophy is correlated with poor verbal fluency in elderly women with stroke, but not in their male counterparts; (3) LAD may be associated with PSD in patients with small subcortical infarcts; (4) MBs in the thalamus are associated with PSEL; (5) frontal lobe infarction and diabetes may be risk factors of insomnia symptoms in stroke patients; and (6) FLA in ischemic stroke may be associated with SVD. (Abstract shortened by UMI.) / Chen, Yangkun. / Adviser: Wai Kwong Tang. / Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 217-238). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Ho, Tik Shun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 88-102). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgement --- p.vii / Table of Contents --- p.viii / List of Table --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xiii / Chapter CHAPTER 1 --- General Introduction --- p.1 / Chapter 1.1 --- Fatty Acid Synthase (FAS) - 7-domain multifunctional enzyme --- p.1 / Chapter 1.1.1 --- Functions --- p.1 / Chapter 1.1.2 --- Structure --- p.2 / Chapter 1.2 --- Fatty Acid biosynthesis reactions --- p.4 / Chapter 1.3 --- Malonyl Coenzyme A - An important mediator in lipogenesis --- p.7 / Chapter 1.4 --- FAS expression in different histotypes --- p.8 / Chapter 1.4.1 --- FAS in normal cells --- p.8 / Chapter 1.4.2 --- FAS in pathological cells --- p.8 / Chapter 1.4.3 --- Tumor-associated FAS (Oncogenic antigen-519) in cancer cells --- p.9 / Chapter 1.5 --- FAS signaling models in breast and prostate cancers --- p.12 / Chapter 1.5.1 --- Association between FAS and PI3K／Akt pathway --- p.12 / Chapter 1.5.2 --- Hypothetical model of FAS hyperactivity in breast and prostate cancer cells --- p.13 / Chapter 1.6 --- FAS inhibition to tackle cancer cell growth --- p.15 / Chapter 1.6.1 --- FAS inhibitors --- p.15 / Chapter 1.6.1.1 --- Cerulenin --- p.16 / Chapter 1.6.1.2 --- C75 --- p.17 / Chapter 1.6.2 --- Small interfering RNA --- p.17 / Chapter 1.7 --- FAS inhibition to enhance chemoresistant cancer cells sensitivity to drugs --- p.19 / Chapter 1.8 --- Hypothesis --- p.20 / Chapter CHAPTER 2 --- Methods and Materials --- p.21 / Chapter 2.1 --- Chemicals and antibodies --- p.21 / Chapter 2.2 --- Cell cultures --- p.21 / Chapter 2.3 --- MTT assay --- p.22 / Chapter 2.4 --- 5-Bromo-2'-deoxyuridine (BrdU)-labeling cell proliferation assay --- p.22 / Chapter 2.5 --- Cytotoxicity detection assay of LDH release --- p.23 / Chapter 2.6 --- DNA flow cytometry --- p.23 / Chapter 2.7 --- Confocal micocropy --- p.24 / Chapter 2.8 --- Immunoblot analysis --- p.24 / Chapter 2.8.1 --- Preparation of protein lysates --- p.24 / Chapter 2.8.2 --- Immunoblotting --- p.25 / Chapter 2.9 --- Caspase inhibitor studies --- p.26 / Chapter 2.10 --- Analysis of mitochondrial membrane potential --- p.26 / Chapter 2.11 --- Determination of caspase activities --- p.27 / Chapter 2.12 --- siRNA transfection --- p.27 / Chapter 2.13 --- Statistical analysis --- p.28 / Chapter CHAPTER 3 --- Results --- p.29 / Chapter 3.1 --- Cytostatic & cytotoxic studies of FAS inhibitors on human cancer cells --- p.29 / Chapter 3.1.1 --- Cerulenin and C75 suppress cell growth of different cancer histotypes --- p.29 / Chapter 3.1.2 --- Cerulenin and C75 suppress cell growth of A-375 dose- and time-dependently --- p.32 / Chapter 3.1.3 --- Cerulenin and C75 exert cytotoxic effect on A-375 but not normal skin HS68 cells --- p.36 / Chapter 3.1.4 --- Cerulenin and C75 arrest cell cycle progression and induce apoptosis with DNA Fragmentation --- p.39 / Chapter 3.2 --- Mechanistic studies of FAS inhibitors in A-375 cells --- p.46 / Chapter 3.2.1 --- Cerulenin and C75 induce caspase-dependent apoptosis --- p.46 / Chapter 3.2.2 --- Cerulenin- and C75-induced apoptosis involve extrinsic death receptor pathway --- p.52 / Chapter 3.2.3 --- Cerulenin- and C75-induced apoptosis involve intrinsic mitochondrial pathway --- p.57 / Chapter 3.2.4 --- Extrinsic death receptor pathway serves as a pioneer and links with intrinsic mitochondrial pathway in cerulenin- and C75-induced apoptosis --- p.65 / Chapter 3.3 --- Small interfering RNA on Fatty Acid Synthase (FAS siRNA) --- p.68 / Chapter 3.3.1 --- FAS siRNA induces PARP cleavage --- p.68 / Chapter 3.3.2 --- FAS siRNA triggers caspase-dependent apoptosis as FAS inhibitors --- p.70 / Chapter CHAPTER 4 --- Discussion --- p.72 / Chapter CHAPTER 5 --- Future Prospect --- p.85 / Chapter CHAPTER 6 --- References --- p.88

O objetivo desse estudo foi avaliar o comprometimento de pequenas vias aéreas na Fibrose Pulmonar Idiopática (FPI) analisando curvas Pressão- Volume (PV) do sistema respiratório. Coletamos curvas PV de doze pacientes antes da biopsia pulmonar, que confirmou FPI em sete pacientes e Pneumonite de Hipersensibilidade em cinco. Todas as curvas foram ajustadas com modelo sigmóide, V = a + b / (1 + e -(P-c/d)), e exponencial V = A - B . e -k.P (aplicado apenas à parte superior). O modelo exponencial, apesar do bom ajuste à parte superior, não representou a parte inicial da curva, gerando parâmetros sem significado. O modelo sigmóide ajustou bem toda a curva e gerou parâmetros com significado fisiológico, que sugerem a presença de colapso de pequenas vias aéreas na FPI. / The objective of this study was to evaluate small airways compromise in Idiopathic Pulmonary Fibrosis (IPF) using pressure-volume (PV) curves of the respiratory system. We collected PV curves from twelve patients before lung biopsy, which confirmed IPF in seven patients and Hipersensitivity Pneumonia in five. All curves were fitted with a sigmoid model, V = a + b / (1 + e -(P-c/d)), and an exponential model, V = A - B . e -k.P (applied only to the superior part of the curve). The exponential model, despite having a good fit to the superior part of the curve, did not represent the initial part, and yielded parameters with no physiological meaning. The sigmoid model had a good fit to the entire curve and yielded parameters with physiological meaning, suggesting the presence of small airways collapse in IPF.

A retroalimentação por eletromiografia é uma técnica de aprendizado para controle voluntário de respostas fisiológicas pelo condicionamento operante. Compararam-se os sinais eletromiográficos dos músculos quadríceps de duas séries de retroalimentação pelo "Método Brucker" com duração de 50 minutos semanais durante quatro semanas com um intervalo de três meses em 20 pacientes com trauma raquimedular cervical na posição sentado e na transição da posição sentado para a ortostática. Demonstrou aumento amplitude do potencial eletromiográfico na segunda série. Comprovou que na transição de sentado para a posição ortostática consegue-se arregimentar um maior número de fibras do quadríceps do que na sentado / Electromyography biofeedback is a learning technique for voluntary control of physiological responses through operant conditioning. The electromyographic signals from the quadriceps muscles of two biofeedback series were compared using the Brucker method, with a 50-minute weekly session during four weeks and a three-month interval, in 20 patients with cervical spinal cord injury, in the sitting position and during the transition from the sitting to the orthostatic position. An increase in the amplitude of the electromyographic potential was shown in the second series. It was proven that during the transition from the sitting to the orthostatic position it is possible to gather a larger number of fibers of the quadriceps as compared to the sitting position