Vários materiais têm sido injetados ou inseridos nas pregas vocais na tentativa de solucionar a incompetência ou insuficiência glótica. Contudo, poucos são os estudos que avaliam o processo cicatricial decorrente da enxertia destes materiais. O objetivo desta pesquisa foi quantificar e comparar as fibras colágenas no músculo vocal das laringes dos coelhos que foram submetidos a enxerto unilateral de gordura ou fáscia muscular com a prega vocal contra-lateral, que foi submetida ao mesmo procedimento, com exceção da enxertia. Estudamos 24 coelhos, divididos em dois grupos com 12 coelhos em cada um. No primeiro grupo, denominado F (Fáscia), os coelhos foram submetidos à inserção de enxerto de fáscia lata autóloga na prega vocal direita. No outro grupo, denominado G (Gordura), os coelhos foram submetidos a implante de gordura autóloga \"em bloco\" também na prega vocal direita. Todos os coelhos foram também submetidos ao mesmo procedimento na prega vocal esquerda, com exceção da colocação do enxerto. A prega vocal esquerda, desta forma, constituiu o \"controle\" para cada coelho. Metade dos coelhos, de cada um dos grupos (F e G), foi sacrificada após 90 dias do procedimento cirúrgico. A outra metade dos coelhos dos grupos G e F foi sacrificada após 180 dias do procedimento cirúrgico. As laringes foram removidas e as pregas vocais, direita e esquerda, foram preparadas histologicamente. Os cortes corados pelo método da picrosírius-polarização foram utilizados para a visualização e análise das fibras colágenas. O colágeno foi analisado morfometricamente através do método da Picrossírius-polarização com a utilização do software Image Pro Plus. Houve aumento do colágeno em todos grupos enxertados quando comparados com o grupo controle. A concentração do colágeno encontrada nos coelhos submetidos a enxerto de gordura foi significativamente maior quando comparados à concentração do colágeno nos coelhos submetidos a enxerto de fáscia muscular, tanto com 90 quanto com 180 dias. A enxertia de gordura e fáscia lata na prega vocal de coelho promoveu maior deposição de colágeno do que no grupo controle, sendo mais exuberante na inserção de gordura / Several materials have been injected or inserted in the vocal fold, in attempt to solve glottic insufficiency. Nevertheless, there is just a few papers that evaluates the inflammatory process resulted from these materials incorporation. The objective of this study was to quantify and compare the collagen fibers in the vocal muscle of the larynx from the rabbits in which unilateral fat and muscular fascia were introduced, to the contralateral vocal fold, in which the same procedure have taken place, except be the grafting. Twenty four rabbits were used in this study, divided into two groups, 12 rabbits each. In the first group, named F (from Fascia), the rabbits underwent the insertion of fascia lata into the right vocal fold. In the other group, named G (from Grease), the rabbits underwent implantation of autologous fat \"en bloc\" in the right vocal fold, as well. All rabbits have undergone the same procedure in the left vocal fold, except for the graft insertion. The left vocal fold, therefore, formed the control group for each rabbit. Half the rabbits, form each group (F and G), was sacrificed after 90 days, while the other half was sacrificed after 180 days from the surgical procedure. The larynxes were removed and the vocal folds, right and left, were prepared for histology, using the method of picrosirius-polarization for the coloration. The collagen fibers in the samples were analyzed using a computer software called Image Pro Plus. An increasing of the collagen was found in all the groups in which grafts have been placed, when compared to the control group. The collagen density found in those rabbits which underwent fat insertion was significantly higher than in those with muscular fascia insertion, for both periods of 90 and 180 days, as well. Fat and muscular fascia insertion in rabbits vocal fold resulted in a higher collagen deposition, when compared to the control group, being

Foi realizado um estudo retrospectivo, por meio da revisão de 72 prontuários de mulheres com diagnóstico de câncer de mama, submetidas ao tratamento quimioterápico neoadjuvante com epirrubicina e docetaxel e no adjuvante, epirrubicina e ciclofosfamida . Os prontuários revisados foram de mulheres na faixa de 30 a 60, acompanhadas no Ambulatório de Mastologia do Departamento de Ginecologia e Obstetrícia do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (HCFMRP-USP) e que receberam o tratamento quimioterápico entre os anos de 2003 e 2006. Resultados: As participantes foram divididas em dois grupos, sendo um das 31 mulheres que apresentaram neutropenia e o outro das 41 que não apresentaram. A média de idade das participantes foi de 47,8 anos. Entre as toxicidades gastrointestinais durante a neoadjuvância e a adjuvância observouse a mucosite (8,4% e 2%), náusea (18,6% e 18%) e vômito (3,3% e 18%). Outra intercorrência observada foi o extravasamento durante o tratamento quimioterápico que ocorreu em 17 (23,6%) mulheres. Observou-se que 43% das mulheres apresentaram neutropenia, que analisadas entre os ciclos de quimioterapia foram estatisticamente significantes para os ciclos dois e três da neoadjuvância com valores de p de 0,0016 e 0,0009 respectivamente, para os ciclos dois e três da adjuvância com valores de p de 0.0014 e 0.0030 respectivamente, para o final do tratamento neoadjuvante, anterior ao tratamento cirúrgico sendo o p-valor=<0.0001 e para o final do tratamento adjuvante, com p-valor=<0.0004. Quanto à ocorrência de anemia, esta não esteve relacionada com a presença ou não de neutropenia, entretanto observou-se que houve uma queda nos valores de HB durante a neoadjuvância, com ligeira recuperação no período de adjuvância, porém, não houve recuperação aos valores médios anteriores ao tratamento quimioterápico. A redução da dose foi utilizada para seis mulheres em decorrência da toxicidade hematológica. Registrou-se 152 atrasos entre os ciclos de quimioterapia. Realizado o teste do Log-Rank para o tempo de tratamento e de sobrevida, concluiu-se que esta foi igual para os dois grupos de mulheres. Conclusão: Por meio dos resultados deste estudo demonstra-se a necessidade de elaboração e implementação de protocolos de cuidados de enfermagem para pacientes oncológicos com a finalidade de avaliação dos eventos adversos e manejo mais adequado dos mesmos / Method: Thais study data were collected retrospectively reviewing the chart of 72 women with breast cancer, underwent to chemotherapy for the first time, that used epirubicin and docetaxel to neoadjuvant treatment and epirubicin and ciclophosphamid to adjuvant treatment. The data collection was done with the charts of women, with 30 to 60 years, treated in 2003 to 2006 in followed in the onco-gynecology and mastology sector- Gynecology and Obstetric Department of the University of São Paulo at Ribeirão Preto Medical School Hospital das Clínicas. Results: The participants had been divided in two groups, one with 31 women who had presented neutropenia and the other with 41 that had not presented. The average of age of the participants was of 47,8 years. The gastrointestinal toxicities during the neoadjuvant and adjuvant treatment observed mucositis (8.4% and 2%), nausea (18.6% and 18%) and vomiting (3.3% and 18%). Another observed toxicity was the extravasation during the chemotherapy treatment that occurred in 17 (23.6%) women. Was observed that 43% of the women had respectively presented neutropenia, who analyzed between the chemotherapy cycles had been statistical significant for cycles two and three of the neoadjuvant with values of p = 0,0016 and 0,0009 respectively, for cycles two and three of the adjuvant with values of p =0.0014 and 0.0030. And for the end of the neoadjuvant treatment, previous treatment to the surgical treatment being p-valor=< 0,0001 and for the end of the adjuvant treatment, with p-valor=< 0.0004. To anemia occurrence, this was not related with the presence or not of neutropenia, however it was observed that had a fall in the values of HB during the neoadjuvant, with fast recovery in the period of adjuvant. However, did not have recovery to previous the average values to the chemoterapy treatment. The reduction of the dose was used for six women in result of the hematologic toxicity. Was registered 152 doses delays between the chemotherapy cycles. The Log-Rank test for the time of treatment and survival, concluded that was equal for both groups. Conclusion: Through the results of this study demonstrates the necessity of develop and implement protocols for nursing care to women with breast cancer in order to assess the adverse events and most appropriate management of them

Introdução. A varfarina tem sido considerada a principal terapêutica anticoagulante oral há aproximadamente 50 anos, estando entre os dez medicamentos mais envolvidos com reações adversas a medicamentos (RAM), apresenta estreita janela terapêutica e complexo regime posológico, exibe enorme variabilidade doseresposta e elevado risco de interações medicamentosas (IM). Objetivo. Identificar e avaliar as IM e RAM relacionadas com a administração da varfarina. Casuística e Métodos. Trata-se de um estudo transversal. Os dados foram coletados retrospectivamente através do banco de dados informatizado do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo pertencente ao Sistema Único de Saúde. As prescrições de janeiro/2004 a dezembro/2010 dos pacientes que utilizaram varfarina foram analisadas, sendo os pacientes divididos em dois grupos: estudo (uso de vitamina K até 168 horas após prescrição de varfarina) e controle. Posteriormente, as prescrições medicamentosas que não continham varfarina foram excluídas da análise. As informações coletadas incluíram idade, gênero, raça, unidade de atendimento, diagnóstico clínico, doses e medicamentos, e exames laboratoriais. As IM com varfarina foram classificadas em risco A, B, C, D e X de acordo a base de dados da Lexi-Interact(TM) Online. Foi realizada análise descritiva e analítica (p<0,05). Resultados e Discussão. Foram identificados 3048 pacientes, os quais receberam 154161 prescrições medicamentosas (42120 continham varfarina). A idade média foi de 55,8 (±19,3) anos, 53,2% do gênero feminino, prevalência de idosos (48,1%) e do diagnóstico outras doenças cerebrovasculares específicas (4,3%). Os valores médios da international normalized ratio (INR) (2,4±1,7) e dose de varfarina (5,1±1,8mg) encontraram-se dentro dos preconizados pelos protocolos terapêuticos. Foi observado que 66,4% dos pacientes realizaram uso de polifarmácia, o que pode elevar o risco de IM. Além disso, 63,2% dos pacientes apresentaram prescrição(ões) de medicamentos classificados como risco D e/ou X, com média de 1,4 (±0,4) medicamento/prescrição, destacando-se o ácido acetilsalicílico e amiodarona. Quando comparado grupo de estudo (n=429) versus controle (n=2619), houve diferença estatisticamente significativa na idade média (anos) (59,0±18,8 vs. 55,5±19,3; p<0,000), número médio de medicamentos/prescrição (7,1±2,8 vs. 6,2±2,8; p<0,000), número médio de medicamentos de Risco D e X de IM por prescrição (1,4±1,3 vs. 1,0±1,0; p<0,000), albumina sérica (g/dL) (3,4±0,6 vs. 3,7±0,6; p<0,000), aspartato aminotransferase (U/L) (60,7± 200,6 vs. 41,5±84,5; p<0,005) e INR (4,9±3,4 vs. 2,1±0,7; p<0,000), fatores estes que podem ter contribuído para ocorrência de RAM no grupo de estudo. Conclusão. Observou-se elevada ocorrência de possíveis IM e RAM nos usuários de varfarina, as quais podem comprometer a efetividade e segurança do tratamento farmacológico. Como possíveis fatores de risco para ocorrência de RAM, destacam-se elevados valores de idade, número de medicamentos/prescrição, prescrição de medicamentos classificados como risco D e/ou X, de INR e de aspartato aminotransferase, e valores diminuídos de albumina sérica. / Introduction. Warfarin has been considered the main oral anticoagulant therapy about 50 years ago and is among the ten drugs most commonly involved in adverse drug reactions (ADR), has a narrow therapeutic index and complex dosage regimen, exhibits enormous variability dose-response and high risk drug-drug interactions (DDI). Objective. To Identify and evaluate DDI and ADR related to the administration of warfarin. Casuistry and Methods. This was a cross sectional study. Data were collected retrospectively through the computerized database of the Faculty of Medicine of Ribeirao Preto Hospital, University of Sao Paulo linked to the Unified Health System. The prescriptions of the January/2004 to December/2010 of patients using warfarin were analyzed, and the patients were divided into two groups: study (utilization of vitamin K until 168 hours after prescribing warfarin) and control. Thereafter, the drug prescriptions that did not contain warfarin were excluded from analysis. Information collected included age, gender, race, patient service center, clinical diagnosis, dosages and drugs, and laboratory exams. The warfarin DDI were classified at risk A, B, C, D and X according to the database Lexi-Interact (TM) Online. Descriptive and analytical analysis were performed (p<0.05). Results and Discussion. We identified 3048 patients who received 154,161 drug prescriptions (42,120 contained warfarin). The mean age was 55.8 (±19.3) years, 53.2% female, prevalence of elderly (48.1%) and other cerebrovascular diseases specific diagnosis (4.3%). The average values of international normalized ratio (INR) (2.4±1.7) and warfarin dose (5.1±1.8mg) were within those recommended by therapeutic protocols. It was observed that 66.4% of patients received polypharmacy, which can raise the risk of DDI. In addition, 63.2% of patients had prescription(s) of drugs classified as D or X risk, with an average of 1.4 (±0.4) drugs per prescription, especially aspirin and amiodarone. Compared study group (n=429) versus control (n =2619), there was a statistically significant difference in mean age (years) (59.0±18.8 vs. 55.5±19.3; p<0.000), average number of medications/prescriptions (7.1±2.8 vs. 6.2±2.8; p<0.000), mean number of drugs with risk D and X DDI/prescription (1.4±1.3 vs. 1.0±1.0, p<0.000), serum albumin (g/dL) (3.4±0.6 vs. 3.7±0.6; p<0.000), aspartate aminotransferases (U/L) (60.7±200.6 vs. 41.5±84.5; p<0.005) and INR (4.9±3.4 vs. 2.1±0.7; p<0.000), factors that may have contributed to the occurrence of ADR in the study group. Conclusion. There was a high occurrence of possible DDI and ADR in patients treated with warfarin, which may compromise the effectiveness and safety of pharmacological treatment. Noteworthy is the high values of age, number of medications/prescriptions, prescription drugs classified as risk D or X, INR and aspartate aminotransferases, and lower values of serum albumin as potential risk factors for the occurrence of ADR.

O processo de administração de medicamentos é considerado um processo complexo, crítico e de alto risco para os pacientes e tem apresentado altas taxas de ocorrência de eventos adversos que poderiam ser evitados. Este estudo teve o objetivo de analisar os riscos potenciais do processo de administração de medicamentos antiinfecciosos por via intravenosa de uma unidade de internação, visando a prevenir e a reduzir eventos adversos com medicamentos. A investigação, de natureza exploratória, foi realizada na unidade de Clínica Médica de um Hospital do Estado de Goiás, utilizando o Método de Análise do Modo e Efeito da Falha. Participaram do estudo, além da pesquisadora, seis profissionais envolvidos na terapêutica medicamentosa: médico, enfermeiro, técnico de enfermagem, farmacêutico e os gerentes de Enfermagem e de risco. Foram realizadas 24 reuniões, no período de 19 de fevereiro a 3 de julho de 2008, totalizando 56 horas. Todas os dados foram transcritos e armazenados em um banco eletrônico no programa Microsoft Excel® e analisados no software XFMEA 4. Os resultados indicaram que o processo de administração de medicamentos possui quatro microprocessos, dez atividades e 22 funções. No processo foram identificados 52 modos potenciais da falha (MPF), sendo que as maiores freqüências estiveram nas atividades de administração de medicamentos (16; 30,8%), preparo de medicamentos (12; 23,1%), aprazamento de medicamentos (5; 9,6%) e transcrição de medicamentos para etiquetas (5; 9,6%). Também foram identificados 79 efeitos potencias da falha (EPF), com as maiores freqüências nas atividades: administração de medicamentos (24; 30,4%), preparo de medicamentos (15; 19%) e transcrição de medicamentos para etiquetas (12; 15,2%). Dos EPF, 36,2% foram considerados de gravidade média; 28,7% de moderada, e 27,5% de alta. Em 80% das atividades, foram identificados efeitos de alta gravidade. A classificação dos efeitos apontou que os tipos mais freqüentes foram os erros de: técnica (21; 26,6%), omissão (20; 25,3%) e horário (15; 19%). Foram identificadas 285 causas potenciais da falha (CPF) com as seguintes freqüências quanto aos índices de ocorrência: 91 (31,9%) média, 78 (27,4%), baixa ou relativamente baixa; 40 (14,0%), alta; e 30 (10,5%), extremamente alta. As CPF foram classificadas dentro de três categorias: gestão dos processos organizacionais (125; 43,9%); recursos humanos (124; 41,4%); estrutura física e material (36; 12,6%). Em relação aos tipos de controles, os resultados mostraram que 211 (92,9%) eram de detecção e 12 (5,3%) de prevenção. O cálculo do número de prioridade de risco (NPR) das CPF identificou que 59 (20,7%) eram de alta prioridade de risco, 156 (54,7%) de média e 70 (24, 6%) de baixa. Foram recomendadas 293 ações de melhorias para as 215 CPF de alta e média prioridade, sendo 240 (81,9%) de curto prazo, 39 (13,3%) de médio prazo e 14 (4,8%) de longo prazo. A simulação do impacto das ações propostas possibilitou identificar uma redução de 79,7% dos MPF de alta prioridade de risco e de 59,6% dos MPF de alta criticidade, assim como uma redução do NPR total das atividades entre 90 e 31,8%, com medidas simples e de rápida aplicação, aumentando a confiabilidade e segurança do processo de administração de medicamentos / Intravenous drug administration is a high-risk process due to its complexity and high rates of adverse events. The aim of this study was to analyse potential risks associated to intravenous anti-infectious drug administration process in a hospital unit. It was an exploratory search at a University Hospital Medical Clinic unit in Goiás, by means of failure modes and effects analysis method. For data collection, it was formed a six members multidisciplinary staff: risk and nurse manager, medical, nurse, and pharmacist, in addition to the searcher. A number of 24 meetings was done, from February 19 and July 3, 2008, in an amount of 56 hours. One has collected data, copied and saved them in a Microsoft Excel® electronic data bank. Afterwards, they were analyzed by means of XFMEA 4 software. Results showed that administration process involves 4 micro process, 10 activities, 22 functions. The search identified 52 failure potential modes (FPM) whose most significant frequencies happened in the following activities: drug administration (16; 30.8%); drug preparation (12; 23.1%); drug delay (5; 9,6%) and drug names transcription to tags (5; 9.6%). The study identified also 79 failure potential effects (FPE), whose higher frequencies were: drug administration (24; 30.4%), drug preparation (15; 19%) and transcription to tags (12; 15.2%). Among FPE, 36.2% were considered as of medium severity ones; 28.7% moderate severity, and 27.5% of high severity ones. High severity effects were identified in 80% of the activities. Effect classification pointed that the most frequent types were the following ones: technical (21; 26.6%), omission (20; 25.3%) and schedule (15; 19%). A number of 285 failure potential causes (FPC) were identified with the following occurrence rates: 91 (31.9%) medium, 78 (27.4%), low or extremely low; 40 (14.0%), high; and 30 (10.5%), extremely high. FPC were classified in three categories: organizational process management (125; 43.9%); human resources (124; 41.4%); physical and material structure (36; 12.6%). Concerning to control types, results showed 211 (92.9%) derived from detection and 12 (5.3%) were prevention ones. FPC Risk priority number (RPN) calculation showed that 59 (20.7%) had high-risk priority; 156 (54.7%) medium and 70 (24.6%) low-risk priority. A number of 293 recommendations were done to high and medium priority FPC: 240 (81.9%) short term, 39 (13.3%) de medium term and 14 (4.8%) de long-term ones. Action impact simulation on failure modes allowed to identify a 79.7% reduction in high priority FPM as well a 59.6% one in high criticality FPM ones by means of simple and quick application measures that can improve reliability and safety in drug administration process

Leung Bo-Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 148-159). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.v / List of Abbreviations --- p.vi / List of Tables --- p.viii / List of Figures --- p.ix / Chapter 1 --- Introduction / Chapter 1.1 --- Food colorants --- p.1 / Chapter 1.2 --- Caramel --- p.3 / Chapter 1.2.1 --- Classes of caramel --- p.3 / Chapter 1.2.2 --- Toxicological studies of caramel --- p.5 / Chapter 1.3 --- Castanea mollissima --- p.9 / Chapter 1.4 --- Antioxidants --- p.10 / Chapter 1.4.1 --- Background --- p.10 / Chapter 1.4.2 --- Methods used to evaluate the antioxidative activity --- p.12 / Chapter 1.4.2.1 --- DPPH* scavenging method --- p.13 / Chapter 1.4.2.2 --- High performance liquid chromatography (HPLC) --- p.13 / Chapter 1.5 --- Microtox® test --- p.19 / Chapter 1.6 --- Mutatox® test --- p.19 / Chapter 1.7 --- Methods used to evaluate the functions of major organs --- p.20 / Chapter 1.7.1 --- Liver --- p.20 / Chapter 1.7.2 --- Kidneys --- p.23 / Chapter 1.8 --- Toxicology --- p.25 / Chapter 1.8.1 --- Acute toxicity test --- p.25 / Chapter 1.8.2 --- Chronic toxicity test --- p.26 / Chapter 1.9 --- Objective --- p.27 / Chapter 2 --- Materials and Methods --- p.28 / Chapter 2.1 --- Plant materials --- p.28 / Chapter 2.2 --- Sample preparation --- p.28 / Chapter 2.3 --- Pigment characterization --- p.30 / Chapter 2.3.1 --- Stability test --- p.30 / Chapter 2.3.2 --- HPLC separation of CP --- p.31 / Chapter 2.3.3 --- Determination of antioxidative activity with the DPPH* scavenging method --- p.31 / Chapter 2.4 --- Microtox® test --- p.33 / Chapter 2.5 --- Mutatox® test --- p.34 / Chapter 2.6 --- Acute toxicity test --- p.35 / Chapter 2.6.1 --- Animals --- p.35 / Chapter 2.6.2 --- Housing and maintenance --- p.35 / Chapter 2.6.3 --- Experimental design --- p.37 / Chapter 2.6.4 --- Chemicals --- p.39 / Chapter 2.6.5 --- Clinical pathology test --- p.41 / Chapter 2.6.5.1 --- Haematology --- p.41 / Chapter 2.6.5.2 --- Blood chemistry --- p.45 / Chapter 2.6.5.3 --- Urinalysis --- p.55 / Chapter 2.6.6 --- Histological study --- p.57 / Chapter 2.6.7 --- Statistical analysis --- p.57 / Chapter 2.7 --- Chronic toxicity test --- p.59 / Chapter 2.7.1 --- Animals --- p.59 / Chapter 2.7.2 --- Housing and maintenance --- p.59 / Chapter 2.7.3 --- Experimental design --- p.59 / Chapter 2.7.4 --- Chemicals --- p.60 / Chapter 2.7.5 --- Clinical pathology test --- p.61 / Chapter 2.7.5.1 --- Haematology --- p.61 / Chapter 2.7.5.2 --- Blood chemistry --- p.62 / Chapter 2.7.5.3 --- Urinalysis --- p.62 / Chapter 2.7.6 --- Histological study --- p.62 / Chapter 2.7.7 --- Statistical analysis --- p.62 / Chapter 3 --- Results --- p.63 / Chapter 3.1 --- Pigment characterization --- p.63 / Chapter 3.1.1 --- Stability test --- p.63 / Chapter 3.1.2 --- HPLC separation of CP --- p.63 / Chapter 3.1.3 --- Antioxidative activities of CP preparations --- p.63 / Chapter 3.2 --- Microtox® test --- p.65 / Chapter 3.3 --- Mutatox® test --- p.65 / Chapter 3.4 --- Acute toxicity test --- p.66 / Chapter 3.4.1 --- Growth rate --- p.66 / Chapter 3.4.2 --- Food and fluid consumption --- p.66 / Chapter 3.4.3 --- Organ-weight --- p.66 / Chapter 3.4.4 --- Clinical pathology tests --- p.68 / Chapter 3.4.4.1 --- Haematology --- p.68 / Chapter 3.4.4.2 --- Blood chemistry --- p.70 / Chapter 3.4.4.3 --- Urinalysis --- p.76 / Chapter 3.4.5 --- Histological study --- p.76 / Chapter 3.5 --- Chronic toxicity test --- p.77 / Chapter 3.5.1 --- Growth rate --- p.77 / Chapter 3.5.2 --- Food and fluid consumption --- p.77 / Chapter 3.5.3 --- Organ-weight --- p.77 / Chapter 3.5.4 --- Clinical pathology tests --- p.78 / Chapter 3.5.4.1 --- Haematology --- p.78 / Chapter 3.5.4.2 --- Blood chemistry --- p.80 / Chapter 3.5.4.3 --- Urinalysis --- p.82 / Chapter 3.5.5 --- Histological study --- p.82 / Chapter 4 --- Discussion --- p.137 / Chapter 4.1 --- Pigment characterization --- p.137 / Chapter 4.2 --- Toxicological studies of CP --- p.140 / Chapter 5 --- Conclusion --- p.147 / References --- p.148

Xie, Shaohua = 室內燃香與香港華人的鼻咽癌發病風險 : / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 124-153). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese; appendixes includes Chinese. / Xie, Shaohua = Shi nei ran xiang yu Xianggang Hua ren de bi yan ai fa bing feng xian :

by Chan Ping Lung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 108-118). / Abstracts in English and Chinese. / Abstract (in Chinese) --- p.ii / Abstract (in English) --- p.iv / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Tables --- p.xiii / List of Figures --- p.xiv / List of Appendices --- p.xv / List of Abbreviations --- p.xvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Epidemiology of catheter associated infections --- p.1 / Chapter 1.1.1 --- Catheter associated infections --- p.1 / Chapter 1.1.2 --- Risk and mortality of CAI --- p.2 / Chapter 1.1.3 --- Etiology of CAI --- p.3 / Chapter 1.1.3.1 --- Venous catheters --- p.4 / Chapter 1.1.3.2 --- Urinary catheters --- p.4 / Chapter 1.2 --- Pathogenesis of CAI --- p.5 / Chapter 1.2.1 --- Central venous catheters (CVC) --- p.6 / Chapter 1.2.2 --- Urinary catheters --- p.7 / Chapter 1.3 --- Adhesion mechanisms --- p.7 / Chapter 1.3.1 --- Definition of adhesion --- p.7 / Chapter 1.3.2 --- Adhesion mechanism --- p.8 / Chapter 1.3.2.1 --- The phase one --- p.8 / Chapter 1.3.2.2 --- The phase two --- p.10 / Chapter 1.4 --- Catheters --- p.10 / Chapter 1.5 --- Biology of Candida albicans --- p.11 / Chapter 1.5.1 --- Taxonomy of Candida albicans --- p.11 / Chapter 1.5.2 --- Morphology --- p.12 / Chapter 1.5.3 --- Genome --- p.13 / Chapter 1.5.4 --- Biology of Candida albicans cell wall --- p.14 / Chapter 1.5.4.1 --- Constituting molecules of Candida albicans cell wall --- p.14 / Chapter 1.5.4.2 --- Organization of Candida albicans cell wall --- p.15 / Chapter 1.6 --- Agglutinin like sequence gene family --- p.16 / Chapter 1.6.1 --- Gene structure of agglutinin like sequence genes --- p.16 / Chapter 1.6.2 --- Sequence similarity --- p.17 / Chapter 1.6.3 --- Sequence variability --- p.18 / Chapter 1.6.4 --- Expression of ALS genes --- p.19 / Chapter 1.6.5 --- The Als proteins --- p.20 / Chapter 1.6.6 --- Functions of Als proteins --- p.21 / Chapter 1.7 --- Adhesion assay --- p.23 / Chapter 1.7.1 --- Adhesion model --- p.24 / Chapter 1.7.2 --- Factors affecting static adhesion model --- p.25 / Chapter 1.7.3 --- Quantitation methods of adherent cells --- p.27 / Chapter 1.7.3.1 --- Sonication --- p.27 / Chapter 1.7.3.2 --- Staining methods --- p.28 / Chapter 1.7.3.3 --- ATP bioluminescence --- p.28 / Chapter 1.8 --- Research model --- p.29 / Chapter Chapter 2 --- Aim of study --- p.31 / Chapter Chapter 3 --- Materials and Methods --- p.32 / Chapter 3.1 --- Preparation of bacteriological reagents --- p.32 / Chapter 3.2 --- Confirmation of identity of Candida albicans and of Saccharomyces cerevisiae --- p.33 / Chapter 3.3 --- Cell culture of fibroblasts --- p.36 / Chapter 3.3.1 --- Preparation of cell culture reagents --- p.36 / Chapter 3.3.2 --- Recovery of freezing fibroblasts --- p.37 / Chapter 3.3.3 --- Establishment of cell line --- p.37 / Chapter 3.4 --- Preliminary study of adherence of Candida albicans to fibroblasts and to catheters --- p.38 / Chapter 3.4.1 --- Adherence to fibroblasts --- p.38 / Chapter 3.4.1.1 --- Preparation of fibroblasts --- p.38 / Chapter 3.4.1.2 --- Preparation of culture of Candida albicans and of Saccharomyces cerevisiae --- p.39 / Chapter 3.4.1.3 --- Adhesion assay --- p.41 / Chapter 3.4.2 --- Adherence to catheters --- p.42 / Chapter 3.4.2.1 --- Preparation of catheters --- p.42 / Chapter 3.4.2.2 --- Adhesion assay --- p.42 / Chapter 3.5 --- "Confirmation of expression of ALS1, ALS5 smaller allele, and ALS6 of Candida albicans in YPD broth" --- p.44 / Chapter 3.5.1 --- RNA extraction of Candida albicans --- p.45 / Chapter 3.5.2 --- "RT-PCR of ALS1, ALS5 smaller allele, and ALS6" --- p.46 / Chapter 3.5.2.1 --- Primers --- p.46 / Chapter 3.5.2.2 --- RT-PCR --- p.47 / Chapter 3.6 --- Establishment of quantitation system of adhesion assay --- p.49 / Chapter 3.6.1 --- Absorbance measurement of Candida albicans stained with safranin --- p.49 / Chapter 3.6.1.1 --- Preparation of Candida albicans culture --- p.49 / Chapter 3.6.1.2 --- Staining of Candida albicans --- p.50 / Chapter 3.6.1.3 --- Viable count of Candida albicans adhered on the 6-well plate --- p.51 / Chapter 3.6.2 --- ATP bioluminescence --- p.52 / Chapter 3.7 --- Effect of inoculum size on adhesion to catheters --- p.53 / Chapter 3.7.1 --- Preparation of adhesion chambers --- p.53 / Chapter 3.7.2 --- Preparation of catheters --- p.54 / Chapter 3.7.3 --- Preparation of Candida albicans culture --- p.54 / Chapter 3.7.4 --- Adhesion assay --- p.55 / Chapter 3.8 --- "Transformation of Saccharomyces cerevisiae with ALS1, ALS5 smaller allele, and ALS6" --- p.57 / Chapter 3.8.1 --- DNA extraction of Candida albicans --- p.58 / Chapter 3.8.2 --- "PCR of ALS1, ALS5 smaller allele, and ALS6" --- p.59 / Chapter 3.8.3 --- Gel extraction --- p.60 / Chapter 3.8.4 --- Restriction digestion of PCR products of ALS genes and cloning plasmids --- p.61 / Chapter 3.8.5 --- "Ligation of ALS1, ALS5 smaller allele, ALS6 with pYES6CT cloning plasmids" --- p.62 / Chapter 3.8.6 --- Transformation of ligated plasmid into Escherichia coli --- p.63 / Chapter 3.8.7 --- Miniprep of plasmids --- p.64 / Chapter 3.8.8 --- DNA sequencing --- p.65 / Chapter 3.8.9 --- Transformation of Saccharomyces cerevisiae --- p.66 / Chapter 3.8.10 --- "Detection of Alsl，Als5, and Als6 protiens expression" --- p.68 / Chapter 3.8.10.1 --- Preparation of cultures in SC synthetic medium --- p.68 / Chapter 3.8.10.2 --- Protein extraction --- p.69 / Chapter 3.8.10.3 --- Dot blot of cell wall lysates --- p.69 / Chapter 3.9 --- Adhesion of transformed Saccharomyces cerevisiae to fibroblasts --- p.71 / Chapter 3.9.1 --- Preparation of fibroblasts and of Saccharomyces cerevisiae cultures --- p.71 / Chapter 3.9.2 --- Adhesion assay --- p.72 / Chapter 3.10 --- "Adhesion of transformed Saccharomyces cerevisiae to FEP, polyurethane, and silicone catheters" --- p.72 / Chapter 3.10.1 --- "Preparation of catheters, adhesion chambers and transformed Saccharomyces cerevisiae cultures" --- p.73 / Chapter 3.10.2 --- Adhesion to catheter fragments --- p.73 / Chapter 3.11 --- Statistical analysis --- p.74 / Chapter Chapter 4 --- Results --- p.75 / Chapter 4.1 --- Confirmation of identity of Candida albicans and of Saccharomyces cerevisiae --- p.75 / Chapter 4.1.1 --- Candida albicans --- p.75 / Chapter 4.1.2 --- Saccharomyces cerevisiae --- p.75 / Chapter 4.2 --- Cell culture of fibroblasts --- p.76 / Chapter 4.3 --- "Preliminary studies of adherence of Candida albicans to fibroblasts and to FEP, polyurethane, and silicone catheters" --- p.76 / Chapter 4.3.1 --- Adherence to fibroblasts --- p.76 / Chapter 4.3.2 --- Adherence to catheters --- p.77 / Chapter 4.4 --- "Confirmation of expression of ALSl, ALS5 smaller allele, and ALS6 of Candida albicans in YPD broth" --- p.78 / Chapter 4.5 --- Establishment of quantitation system of adhesion assay --- p.79 / Chapter 4.5.1 --- Absorbance measurement of Candida albicans stained with safranin --- p.79 / Chapter 4.5.2 --- ATP bioluminescence --- p.79 / Chapter 4.6 --- Effect of inoculum size on adhesion to catheters --- p.80 / Chapter 4.7 --- "Transformation of Saccharomyces cerevisiae with ALS1, ALS5 smaller allele, and ALS6" --- p.81 / Chapter 4.7.1 --- "PCR of ALSl, ALS5 smaller allele, and ALS6" --- p.81 / Chapter 4.7.2 --- Ligation of PCR products with pYES6CT plasmids --- p.82 / Chapter 4.7.3 --- "DNA sequencing results of ALS1, ALS5 smaller allele, and ALS6 ligated plasmids" --- p.83 / Chapter 4.7.4 --- "Detection of Alsl, Als5, and Als6 proteins expression" --- p.84 / Chapter 4.8 --- Adhesion of transformed Saccharomyces cerevisiae to fibroblasts --- p.84 / Chapter 4.9 --- "Adhesion of transformed Saccharomyces cerevisiae to FEP, polyurethane and silicone catheters" --- p.85 / Chapter Chapter 5 --- Discussion --- p.89 / Chapter 5.1 --- Limitations of static adhesion assay model --- p.89 / Chapter 5.2 --- Quantitation System --- p.90 / Chapter 5.2.1 --- Staining method --- p.90 / Chapter 5.2.2 --- ATP bioluminescence assay --- p.91 / Chapter 5.3 --- "Preliminary studies of adherence of Candida albicans to fibroblasts and to FEP, polyurethane, and silicone catheters" --- p.93 / Chapter 5.4 --- Effect of inoculum size on adhesion to catheters --- p.94 / Chapter 5.5 --- Selection of ALS genes --- p.96 / Chapter 5.6 --- Adhesion assay of transformed Saccharomyces cerevisiae to fibroblasts --- p.97 / Chapter 5.7 --- Adhesion assay of transformed Saccharomyces cerevisiae to catheters --- p.99 / Chapter 5.8 --- Alternative research model --- p.101 / Chapter 5.9 --- Implications and future work --- p.102 / Chapter Chapter 6 --- Conclusion --- p.107 / References --- p.108 / Tables --- p.119 / Figures --- p.123 / Appendices --- p.136

To Man Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 85-105). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / (in Chinese) --- p.iv / Acknowledgements --- p.vi / Publications --- p.viii / Table of Contents --- p.ix / List of Tables --- p.xii / List of Figures --- p.xiii / List of Abbreviations --- p.xiv / Chapter CHAPTER 1: --- General Introduction --- p.1 / Chapter Section 1.1 --- Background and Clinical Application of Anthracylines --- p.1 / Chapter Section 1.2 --- DOX-induced Cardiotoxicity --- p.3 / Chapter 1.2.1 --- Types of Cardiotoxicity --- p.4 / Chapter 1.2.1.1 --- Acute Cardiotoxicity --- p.4 / Chapter 1.2.1.2 --- Chronic Cardiotoxicity --- p.5 / Chapter 1.2.2 --- Subcellular Effects of DOX --- p.6 / Chapter 1.2.2.1 --- Ultrastructural Lesions --- p.6 / Chapter 1.2.2.2 --- Effects on Mitochondrial Functions --- p.7 / Chapter 1.2.2.3 --- Effects on Sarcoplasmic reticulum (SR) Functions --- p.8 / Chapter Section 1.3 --- Mechanisms of DOX-induced Cardiotoxicity --- p.8 / Chapter 1.3.1 --- Formation of Free Radicals --- p.9 / Chapter 1.3.1.1 --- Generation of Free Radicals by DOX --- p.10 / Chapter 1.3.1.2 --- Cardiac damage by Free radicals --- p.12 / Chapter 1.3.2 --- Induction of Apoptosis --- p.14 / Chapter 1.3.2.1 --- Characteristics and Pathway of Apoptosis --- p.14 / Chapter 1.3.2.2 --- Mitochondria and Apoptosis --- p.15 / Chapter 1.3.2.3 --- Caspases and Apoptosis --- p.17 / Chapter 1.3.2.4 --- Apoptosis and DOX-induced Cardiotoxicity --- p.18 / Chapter Section 1.4 --- Strategies to Reduce DOX-induced Cardiotoxicity --- p.19 / Chapter 1.4.1 --- Dosage optimization and Schedule modification --- p.19 / Chapter 1.4.2 --- Anthracycline Analogues --- p.21 / Chapter 1.4.3 --- Cardioprotective Agents --- p.21 / Chapter Section 1.5 --- Thrombopoietin --- p.23 / Chapter CHAPTER 2: --- Hypotheses and Objectives --- p.30 / Chapter CHAPTER 3: --- Methodology --- p.33 / Chapter Section 3.1 --- Methods --- p.33 / Chapter 3.1.1 --- Culture of Rat H9C2 Myoblast Cell Line and Primary Neonatal Rat Cardiomyocytes --- p.33 / Chapter 3.1.1.1 --- Maintenance of Cell Line --- p.33 / Chapter 3.1.1.2 --- Culture of Primary Neonatal Rat Cardiomyocytes --- p.34 / Chapter 3.1.2 --- Effects of Thrombopoietin on Cell Viability of Rat H9C2 Myoblast Cell Line and Beating Rates of Primary Rat Cardiomyocytes --- p.35 / Chapter 3.1.2.1 --- Cell Viability assay --- p.35 / Chapter 3.1.2.2 --- Beating Rate of Primary Beating Cardiomyocytes --- p.36 / Chapter 3.1.3 --- Effects of Thrombopoietin on the Protection against DOX-induced Heart Injury In Vivo --- p.36 / Chapter 3.1.3.1 --- Animals --- p.36 / Chapter 3.1.3.2 --- Experimental Protocol --- p.37 / Chapter 3.1.3.3 --- Echocardiography --- p.38 / Chapter 3.1.3.4 --- Blood Cell Counts --- p.39 / Chapter 3.1.3.5 --- Histopathology --- p.39 / Chapter 3.1.4 --- Effects of Thrombopoietin on Apoptosis and Mitochondrial Integrity of Rat H9C2 Myoblast Cell Line and Apoptosis In Vivo --- p.40 / Chapter 3.1.4.1 --- Determination of Externalized Phosphatidylserine --- p.40 / Chapter 3.1.4.2 --- Determination of Active Caspase-3 Expression --- p.41 / Chapter 3.1.4.3 --- Assessment of Mitochondrial Integrity --- p.42 / Chapter 3.1.4.4 --- TUNEL assay --- p.43 / Chapter 3.1.5 --- Statistical Analysis --- p.44 / Chapter CHAPTER 4: --- Effects of Thrombopoietin on Cell Viability of Rat H9C2 Myoblast Cell Line and Beating Rates of Primary Neonatal Rat Cardiomyocytes --- p.46 / Chapter Section 4.1 --- Results --- p.46 / Chapter 4.1.1 --- Effects of TPO on DOX-induced Cell Death --- p.46 / Chapter 4.1.2 --- Effects of TPO on the Beating Rates of Primary Cardiomyocytes --- p.47 / Chapter Section 4.2 --- Discussion --- p.47 / Chapter CHAPTER 5： --- Effects of Thrombopoietin on the Protection Against DOX-induced Heart Injury In Vivo --- p.54 / Chapter Section 5.1 --- Results --- p.54 / Chapter 5.1.1 --- General Observations and Survival --- p.54 / Chapter 5.1.2 --- Blood Cell Counts --- p.55 / Chapter 5.1.3 --- Cardiac Functions by Echocardiography --- p.56 / Chapter 5.1.4 --- Gross Anatomic Changes and Pathology of the Myocardium --- p.57 / Chapter Section 5.2 --- Discussion --- p.58 / Chapter CHAPTER 6: --- Effects of Thrombopoietin on Apoptosis and Mitochondrial Integrity of H9C2 Cell Line and Apoptosis In Vico --- p.69 / Chapter Section 6.1 --- Results --- p.69 / Chapter 6.1.1 --- Determination of Externalized Phosphatidylserine --- p.69 / Chapter 6.1.2 --- Determination of Active Caspase-3 Activity --- p.70 / Chapter 6.1.3 --- Assessment of Mitochondrial Membrane Potential --- p.70 / Chapter 6.1.4 --- Determination of Apoptosis by TUNEL assay --- p.72 / Chapter Section 6.2 --- Discussion --- p.72 / Chapter CHAPTER 7： --- General Discussion and Conclusion --- p.83 / References --- p.85

肺癌是一個世界性的健康難題。大量研究證據顯示，煙草及其致癌物NNK對環氧酶（COX）-2及其下游產物具有促進效應。血栓素（TxA）2是COX-2的關鍵性下游產物之一，該論文闡述了TxA2在NNK導致的肺癌增長中的可能作用。 / 我們發現相对于非吸烟者，吸煙者肺癌組織表达更高水平的TxA2合酶（TxAS）。NNK可以刺激培養的肺癌細胞TxA2合成。用TxAS抑制劑和TxA2受體（TP）拮抗劑分別阻抑TxA2的合成與功能可以引起細胞凋亡，從而有效抑制NNK導致的細胞增殖效應。在TxA2合成受抑制的情況下，TP激動劑U46619幾乎可以重建NNK效應，說明TP在NNK效應中的重要作用。研究還顯示，激活的TP可以通過PI3K/Akt和ERK通路進一步激活CREB，從而參與NNK對肺癌細胞的促生長效應。 / 緊接著，我們的研究顯示TP 可以調節NNK對COX-2 和TxA2的誘導，而且發現NNK刺激的TxA2合成主要依賴於COX-2活性。COX-2和TxA2功能抑製劑對NNK的促細胞生長作用具有相似的抑制效用。考慮到TP是TxA2的功能受體，該資料說明TP在NNK處理的肺癌細胞中傳遞了上游因子COX-2的促腫瘤作用。在使用COX-2小干擾RNA（siRNA）抑制NNK作用的情況下，TP激動劑U46619幾乎可以恢復NNK的效應證實了TP的傳遞者角色。研究還發現 TPα而不是TPβ在培養的肺癌細胞系中廣泛表達，並且過表達TPα具有促進腫瘤生長的作用。在用NNK處理細胞的條件下，TPα還具有促COX-2表達和TxA2生成的作用。 / 我們的研究進一步發現，在吸煙者肺癌組織中TPα表達增高，這與TxAS的表達相似。与此结果相一致，在經NNK處理的A/J小鼠肺癌組織中，TxAS和TP表達水準也是明顯上升的。在細胞培養實驗中，NNK能夠提高TxAS蛋白和信使RNA（mRNA）的表達水準。但是，在TP的兩個亞型TPα和TPβ中， NNK僅能促進TPα的蛋白表達，對它們的mRNA均無影響。NNK對TxAS的促表達作用是核轉錄因數(NF)-κB依賴性的。其他的幾個關鍵轉錄因數，諸如特異性蛋白(SP)-1，CREB和活化受體 （PPAR）γ均未參與NNK對TxAS和TPα的表達促進作用。進一步的，轉錄後機理被證實參與了NNK對TPα的作用。TPα而不是TPβ經鑒別在NNK的促NF-κB 激活 和 促TxAS 表達效應中起正向調節作用。 / 總之， 我們的研究說明TxA2相關通路在NNK的促肺癌細胞生長效應中起正向調節作用。我們的研究揭示了TPα的自我激活環路。通過該環路，TxA2，或者說TxAS和TPα參與了NNK的肺癌促生長效應。因此，我們的研究為肺癌的防治了提供了一個新的方向，即靶向TxAS和TPα是一種可能有效的策略。 / Lung cancer concerns a world-wide health problem. There is considerable evidence of that tobacco smoke and its carcinogen 4-methylnitrosamino-1-3-pyridyl-1-butanone (NNK) have the potential effects on the production of cyclooxygenase (COX)-2 and its downstream products in tumor cells. This thesis is constructed to describe the study focused on the role of thromboxane A2 (TxA2), one of the key downstream products of COX-2, in NNK-induced lung tumor growth. / We found that as compared to non-smokers, lung cancer tissues obtained from smokers tended to express more TxA2 synthase (TxAS). Moreover, NNK could stimulate TxA2 synthesis in lung cancer cells. Blockade of TxA2 synthesis and action by TxAS inhibitor and TxA2 receptor (TP) antagonist completely blocked NNK-promoted cell proliferation via inducing apoptosis. Moreover, TP agonist U46619 reconstituted a near full proliferative response to NNK when TxAS was inhibited, affirming the role of TP in NNK-induced cell growth. Furthermore, we revealed that the activated TP may then activate CREB through PI3K/Akt and ERK pathways, thereby contributing to the NNK-induced lung cancer cell growth. / We subsequently showed that TP could modulate the induction of COX-2 and TxA2 by NNK. The synthesis of TxA2 stimulated by NNK was found to be mainly dependent on COX-2 activity. Intriguingly, there are similar inhibitory effects on NNK-induced cell growth between pharmacological inhibition of COX-2 and the blockade of TxA2 synthesis and action. Because TP is the natural receptor of TxA2, these results suggest that TP may function as a mediator for the tumor-promoting effects of COX-2 upon NNK treatment, which was confirmed by the data showing that U46619 almost restored NNK effects in the presence of COX-2-siRNA. Importantly, TPα, but not TPβ was found to be widely expressed in lung cancer cells and be able to promote tumor growth, COX-2 expression and TxA2 synthesis upon NNK treatment. / We further demonstrated that in lung tumor tissues obtained from smoker, TPα protein was increased, which was similar to the change in TxAS protein. The increased levels of TxAS and TP proteins were also found in lung cancer tissues of A/J mice treated with NNK. In cell culture experiments, NNK could increase TxAS at both protein and mRNA levels. However, TPα rather than TPβ was increased by NNK at protein but not mRNA level. NNK-stimulated TxAS expression was dependent on nuclear factor (NF)-κB signaling. Other key transcriptional factors, such as specificity protein(SP)-1, CREB and peroxisome proliferator-activated receptor-gamma (PPARγ), were not involved in NNK-induced TxAS and TPα expression. Further experiments revealed that post-transcriptional mechanisms were responsible for NNK-induced TPα expression. TPα rather than TPβ was finally identified to have a positive role in NNK-induced NF-κB activation and TxAS expression. / Taken together, our study suggests that TxA2 pathway has a positive role in NNK-induced lung cancer cell growth. An auto-positive feedback loop of TPα activation to facilitate lung tumor growth in the presence of NNK is delineated by these results. Therefore, targeting TxAS or/and TPα may represent a promising strategy for prevention and treatment of lung cancer. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Huang, Runyue. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 119-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.I / 摘要 / Publications / Acknowledgement / Abbreviations / Table of contents / Chapter Chapter 1 --- General introduction--Tobacco smoking, COX-2 pathway and cancer / Chapter 1.1 --- Abstract --- p.1 / Chapter 1.2 --- Introduction --- p.2 / Chapter 1.3 --- Cyclooxygenase and prostanoids --- p.5 / Chapter 1.4 --- The effects of tobacco smoking on COX-2 pathway, and the related pathologies --- p.8 / Chapter 1.4.1 --- Smoking, PGE2, inflammation and immunosupression --- p.8 / Chapter 1.4.2 --- Smoking, TxA2, platelet activation, cell contraction and angiogenesis --- p.11 / Chapter 1.4.3 --- Smoking and PGI2 --- p.16 / Chapter 1.5 --- The role of cyclooxygenase-2 pathway in the progression of tobacco smoke-related cancers --- p.19 / Chapter 1.5.1 --- Lung cancer --- p.19 / Chapter 1.5.2 --- Gastrointestinal cancer --- p.23 / Chapter 1.5.3 --- Bladder cancer --- p.24 / Chapter 1.5.4 --- Head and neck squamous cell carcinoma --- p.25 / Chapter 1.5.5 --- The signaling mechanisms underlying the induction of COX-2 by smoking in tumors --- p.26 / Chapter 1.6 --- Summary, future directions and key questions --- p.28 / Chapter Chapter 2 --- NNK induces lung cancer cell growth by stimulating TxA2 and its receptor / Chapter 2.1 --- Abstract --- p.32 / Chapter 2.2 --- Introduction --- p.33 / Chapter 2.3 --- Materials and Methods --- p.35 / Chapter 2.3.1 --- Cell lines and cell culture --- p.35 / Chapter 2.3.2 --- Chemicals and drug treatment --- p.35 / Chapter 2.3.3 --- Thromboxane B2 EIA assay --- p.36 / Chapter 2.3.4 --- MTT assay --- p.36 / Chapter 2.3.5 --- BrdU cell proliferation assay --- p.37 / Chapter 2.3.6 --- Flow cytometry for analysis of apoptosis --- p.37 / Chapter 2.3.7 --- Transfection of cells with CREB siRNA --- p.38 / Chapter 2.3.8 --- Western blot analysis and antibodies --- p.38 / Chapter 2.3.9 --- Statistical analysis --- p.39 / Chapter 2.4 --- Results --- p.41 / Chapter 2.4.1 --- High expression of TxAS in lung cancer tissues of smoker --- p.41 / Chapter 2.4.2 --- NNK stimulated TxA2 synthesis in lung cancer cells --- p.43 / Chapter 2.4.3 --- Blockade of TxA2 synthesis and action prevented NNK-induced cell growth --- p.44 / Chapter 2.4.4 --- TxA2 mimetic U46619 reconstituted NNK-enhanced cell proliferation under TxA2-inhibited condition --- p.47 / Chapter 2.4.5 --- Blockade of TxA2 synthesis or action induced the apoptosis of the NNK-exposed cells --- p.47 / Chapter 2.4.6 --- CREB is accountable for the key role of TxA2 in NNK-enhanced cell proliferation --- p.49 / Chapter 2.4.7 --- PI3K/Akt and ERK rather than JNK and p38 pathways were mediated by TxA2 in the NNK-exposed cells --- p.52 / Chapter 2.4.8 --- CREB is located downstream of the PI3K/Akt and ERK pathways in NNK-treated cells --- p.53 / Chapter 2.5 --- Discussion --- p.55 / Chapter Chapter 3 --- The positive role of TPα in the induction of COX-2, TxA2 and cell growth by NNK in human lung cancer cells / Chapter 3.1 --- Abstract --- p.62 / Chapter 3.2 --- Introduction --- p.63 / Chapter 3.3 --- Materials and methods --- p.65 / Chapter 3.3.1 --- Cell culture and chemicals --- p.65 / Chapter 3.3.2 --- Transient transfections --- p.66 / Chapter 3.3.3 --- TxB2 measurement --- p.66 / Chapter 3.3.4 --- Cell growth detection --- p.67 / Chapter 3.3.5 --- Analysis of apoptosis --- p.67 / Chapter 3.3.6 --- Western blot analysis and antibodies --- p.67 / Chapter 3.3.7 --- Statistical analysis --- p.68 / Chapter 3.4 --- Results --- p.70 / Chapter 3.4.1 --- Examination of TP as the modulator for induction of COX-2 and TxA2 by NNK --- p.70 / Chapter 3.4.2 --- The TxA2 generated in cells treated with NNK is mainly dependent on COX-2 activity --- p.72 / Chapter 3.4.3 --- Examination of TP as the key mediator for the tumor-promoting effect of COX-2 --- p.72 / Chapter 3.4.4 --- The expression and action of α and β isoforms of TP in human lung cancer cells --- p.77 / Chapter 3.4.5 --- the identification of positive role of TPα in NNK-induced COX-2, TxA2 and cell growth in lung cancer cells --- p.79 / Chapter 3.5 --- Discussion --- p.81 / Chapter Chapter 4 --- TP-α facilitates lung tumor growth through an autoregulatory feedback mechanism / Chapter 4.1 --- Abstract --- p.88 / Chapter 4.2 --- Introduction --- p.89 / Chapter 4.3 --- Materials and methods --- p.91 / Chapter 4.3.1 --- Human lung tissue and immunohistochemical analysis --- p.91 / Chapter 4.3.2 --- Animal treatment --- p.91 / Chapter 4.3.3 --- Cell culture and chemicals --- p.92 / Chapter 4.3.4 --- Transient transfection --- p.93 / Chapter 4.3.5 --- Real-time PCR --- p.93 / Chapter 4.3.6 --- Western blot analysis and antibodies --- p.94 / Chapter 4.3.7 --- Statistical analysis --- p.95 / Chapter 4.4 --- Results --- p.96 / Chapter 4.4.1 --- The effects of smoking on the expression of TP in human lung cancer tissue --- p.96 / Chapter 4.4.2 --- The effects of NNK on the expression of TxAS and TP in lung tissues of A/J mice --- p.98 / Chapter 4.4.3 --- The effects of NNK on the expression of TxAS and TPα in lung cancer cells --- p.99 / Chapter 4.4.4 --- Identification of the roles of NF-κB, CREB and SP1 in NNK-induced TxAS and TPα expression --- p.101 / Chapter 4.4.5 --- The negative role of PPARγ in NNK-induced TxAS and TPα expression --- p.104 / Chapter 4.4.6 --- NNK-induced TPα expression via post-transcriptional mechanism --- p.105 / Chapter 4.4.7 --- Examination of TPα auto-activation mechanism in lung cancer cells stimulated with NNK --- p.106 / Chapter 4.5 --- Discussion --- p.109 / Chapter Chapter 5 --- Conclusion and future works / Chapter 5.1 --- Conclusion --- p.114 / Chapter 5.2 --- Future works --- p.115 / Chapter 5.2.1 --- The possible role of miR-34c in the auto-regulatory loop of TxAS expression or TPα activation --- p.116 / Chapter 5.2.2 --- The possible role of FOXO3a in the auto-regulatory loop of TxAS expression or TPα activation --- p.116 / References --- p.119

Submitted by Rosina Valeria Lanzellotti Mattiussi Teixeira (rosina.teixeira@unisantos.br) on 2015-05-11T12:13:56Z No. of bitstreams: 1 Cristine Galante.pdf: 2179542 bytes, checksum: 271fb9c0732466054cde0ba5f209ecd9 (MD5) / Made available in DSpace on 2015-05-11T12:13:56Z (GMT). No. of bitstreams: 1 Cristine Galante.pdf: 2179542 bytes, checksum: 271fb9c0732466054cde0ba5f209ecd9 (MD5) Previous issue date: 2013-04-08 / INTRODUCTION: The region of Santos and São Vicente estuary covered by the hospitalization for acute diarrhea disease and their correlation with the distribution of water to the population. In the municipalities of Bertioga, Cubatão, Guarújá, Santos e São Vicente there are many isolated areas with neighborhoods that are not served by the local water and sewer company. The local company serves 100% of the regular areas, but the isolated areas don´t receive any service from the company. OBJECTIVES: Evaluating the temporal distribution of severe cases of diarrhea in the region of Baixada Santista between 2000 and 2010. Analiying the pattern of occurrence of diarrhea cases by age group. Evaluating the annual correlation of the hospitalization for diarrhea disease with the standard of water quality. METHODS: Ecological study of temporal sequence of severe cases of diarrhea occurred in five cities. Standard and nonstandard rates were used for the population considering hospitalizations for acute diarrhea diseases by age group. The Linear regression correlation was analyzed between the standardized and nonstandardized rates and water quality. Pearson product ¿ moment correlation coefficient with the hospitalization cases for diarrhea disease and the water quality parameter. RESULTS: Among 6355 cases of hospital admission for acute diarrhea diseases between 2000 and 2010, a seasonal pattern was observed in all municipalities. Bertioga and Cubatão, mainly in the first years of study, showed the most cases of hospitalization, mainly among children and elderly. There was not significant correlation of the indicator of water quality with the hospitalization rate. There was a reduction of hospital admissions for acute diarrhea diseases throughout the studied period with a higher incidence in Bertioga and Cubatão. Children that are more likely to be infected are those und four years of age. CONCLUSION: There was a reduction of cases of acute severe diarrhea disease throughout the studied period. A more visible reduction was observed in Bertioga and a less visible one in Cubatão. / INTRODUÇÃO: A região do Estuário de Santos e São Vicente pelo estudo das internações por doenças diarréicas agudas e sua correlação com a qualidade da água para a população. Nas cidades de Bertioga, Cubatão, Guarujá, Santos e São Vicente existem inúmeras áreas desconformes que não estão entre os bairros onde a companhia de saneamento local atende com água encanada e esgoto. A companhia de saneamento local atende cem por cento das áreas regulares, mas as desconformes estão à própria sorte. OBJETIVOS: Descrever o perfil temporal de casos graves de diarréia nas cinco cidades estudadas na região da Baixada Santista entre 2000 e 2010 e sua correlação com a qualidade da água disponibilizada à população. Analisar o padrão de ocorrência dos casos de diarréia por faixa etária. Avaliar a correlação anual dos casos de internação por diarreia com os padrões de qualidade da água nos municípios analisados. METODOS: Estudo Ecológico de séries temporais dos casos graves de diarréia ocorridos nas cidades de Bertioga, Cubatão Guarujá, Santos e São Vicente. Foi feita a análise de distribuição temporal mensal entre janeiro 2000 e dezembro 2010. Foram calculadas Taxas Padronizadas e Não Padronizadas para a população por faixa etária das internações hospitalares de doenças diarréicas agudas. Foi Analisada a Correlação de Regressão Linear entre as entre as Taxas Padronizadas, Não Padronizadas e a qualidade da água. Coeficiente de Correlação de Pearson entre as taxas de internação hospitalar por doenças diarréicas e os parâmetros de qualidade da água. Modelos de Regressão Linear entre as taxas de internação os parâmetros de qualidade da água. RESULTADOS: Dos 6.355 casos de internações hospitalares por doenças diarréicas agudas entre 2000 e 2010, constatou-se o mesmo padrão sazonal em todos os municípios. Bertioga e Cubatão principalmente nos primeiros anos do estudo apresentaram mais casos de internações principalmente nas faixas etárias infantis e nas mais idosas. Não houve correlação significativa entre os indicadores de qualidade de água e as taxas de internação. Houve uma redução dos casos de internação por doenças diarréicas agudas ao longo do período estudado com incidência maior em Bertioga e Cubatão. As crianças mais susceptíveis são menores de quatro anos. CONCLUSÕES: Houve ao longo do período estudado uma redução por doenças diarréicas aguda sendo esta redução mais percebida em Bertioga e em menor proporção em Cubatão.