Endometriosis is a benign inflammatory disorder, defined by the presence of endometrial tissue outside the uterus with lesions typically found on the pelvic peritoneum in close association with the peritoneal mesothelium. The prevalence of endometriosis is estimated at 6-10% of women of reproductive age and it is associated with chronic pelvic pain, dysmenorrhoea, dyspareunia and infertility. Surgical excision can provide symptom relief, but symptoms recur in up to 75% of surgical cases and available medical treatments have undesirable side effects. New treatments are limited due to our poor understanding of the aetiology of endometriosis. To date, the majority of research has focused on changes within the ectopic endometrial tissue to explain the development of endometriosis lesions, however, there is increasing evidence that the peritoneal mesothelium plays an important role. According to Sampson’s theory of retrograde menstruation, ectopic endometrial cells must first have to attach to the surface of the peritoneum before undergoing invasion, proliferation, and neoangiogenesis. TGF-β1 is an inflammatory growth factor that regulates a variety of cellular functions including; cell adhesion, cell invasion and angiogenesis. Levels of TGF-β1 are increased in the peritoneal fluid of women with endometriosis compared to controls and research using a mouse model of endometriosis has demonstrated TGF-β1-null mice to develop smaller and fewer endometriosis lesions than their wild-type controls. Together these studies suggest that TGF-β1 plays a major role in the development of peritoneal endometriosis lesions and that targeting this pathway may be of therapeutic potential. However the functional role that TGF-β1 plays in peritoneal endometriosis is still unclear. The overall aim of this thesis was to determine if the peritoneal expression of TGF-β and its target genes are disrupted in women with endometriosis and whether this could contribute to the development of endometriosis lesions. Our first aim was to determine if the peritoneum was a source of TGF-β expression and if reception and/or signalling were altered in women with endometriosis. We found that the peritoneal fluid of women with endometriosis contained increased concentrations of TGF-β1 and that peritoneal mesothelial cells adjacent to endometriosis lesions expressed significantly higher levels of TGF-β1 mRNA. Analysis of TGF-β signalling targets within the peritoneum showed that women with endometriosis express significantly higher levels of TGF-β targeted genes associated with tumourigenesis processes including; EMT, invasion and angiogenesis. We next asked if there are changes in the metabolic phenotype of endometriosis lesions and peritoneum in women with endometriosis, similar to the metabolic changes seen in tumour cells. Endometriosis lesions expressed markers of aerobic glycolysis, including HIF-1α, suggesting that lesions may metabolise in a similar fashion to tumours. Furthermore, peritoneum adjacent to endometriosis lesions expressed significantly higher levels of markers of aerobic glycolysis, including HIF-1α, suggesting that the peritoneum may feed forward high-energy lactate, a by-product of glycolysis, to the endometriosis lesions. These observations were supported by significantly increased lactate concentrations within the peritoneal fluid of women with endometriosis that positively correlated with levels of TGF- β1. TGF-β1 was shown to increase expression of glycolysis markers and lactate expression in peritoneal mesothelial cells in-vitro, suggesting TGF-β may regulate this change. We then determined if TGF-β1 was responsible for the change in peritoneal mesothelial cell metabolism by signalling through the ID-HIF-1α pathway. ID proteins are transcription factors, whose expression is regulated by the TGF-β superfamily. We found expression of ID1 mRNA to be increased in the peritoneum of women with endometriosis and that TGF-β1 significantly increased ID1 but decreased ID2 expression in the peritoneal mesothelial cells in-vitro. ID1 siRNA knockdown decreased glycolysis initiator HIF-1α mRNA and ID2 siRNA knockdown increased HIF-1α mRNA and lactate expression, suggesting TGF-β1 regulates mesothelial cell metabolism, at least in part, through the ID pathway. ID transcription factors are also known to regulate VEGF-A expression, therefore we next determined if TGF-β1 induced ID1 and/or reduced ID2 expression in the peritoneum promoted VEGF-A mRNA and protein expression. VEGF-A, a cytokine essential for angiogenesis, was significantly increased in the peritoneal fluid of women with endometriosis and levels positively correlated with TGF-β1. TGF-β1 increased VEGF-A expression in-vitro and siRNA knockdown of ID1 decreased and siRNA knockdown of ID2 increased VEGF-A mRNA and protein expression in the peritoneal mesothelial cell. Lastly we aimed to determine if TGF-β1 induces EMT in peritoneal mesothelial cells. The peritoneum of women with endometriosis expressed higher levels of EMT markers. Exposure of peritoneal mesothelial cells to TGF-β1 in-vitro induced EMT-like changes, including; changes to cell morphology, gene expression and protein localisation. Peritoneal mesothelial cells were more migratory and invasive suggesting that TGF-β1 induced EMT may disrupt the mesothelial cell monolayer allowing ectopic endometrial cells to invade the peritoneal tissue or for peritoneal mesothelial cells to migrate into endometriosis lesions. In summary, the novel data presented in this thesis provides evidence that the pelvic peritoneum and in particular the peritoneal mesothelial cell may play a critical role in the aetiology of peritoneal endometriosis. Expression of TGF-β1 and its transcriptional target genes are dysregulated in the peritoneum of women with endometriosis. TGF-β regulated ID expression may induce changes in cell metabolism and promote neoangiogenesis, prompting peritoneal endometriosis lesion development. Furthermore other TGF-β1 transcriptional targets, such as those involved in EMT, are also altered in the peritoneum of women with endometriosis and may contribute the development and maintenance of lesion formation. These results point to a central role for TGF-β1 expression and signalling in the aetiology of peritoneal endometriosis. Furthermore it is likely that changes within the expression profile and morphology of the peritoneal mesothelial cells contribute to peritoneal lesion formation. Drugs that target these pathways may provide new therapies for women with endometriosis.

Orientador: Paqual Barretti / Coorientador: Maria de Lourdes Ribeiro se Souza da Cunha / Banca: Roberto Pecoits-Filho / Banca: Nilton Lincopan / Banca: Jacqueline Costa Teixeira Caramori / Banca: Augusto Cezar Montelli / Resumo: As peritonites bacterianas se mantêm como a principal complicação do método de diálise peritoneal, sendo as espécies de Staphylococcus os agentes mais frequentes destas infecções. As infecções peritoneais devidas à espécie S. aureus apresentam pior evolução clínica, contrastando com a maior resistência antimicrobiana apresentada pelos estafilococos coagulase negativa. As características do agente, associadas às condições do hospedeiro, devem, portanto, estar relacionadas à evolução de um episódio. Assim, o objetivo deste trabalho foi avaliar a influência do perfil clonal, dos fatores de virulência e da resistência antimicrobiana de Staphylococcus, juntamente com fatores do hospedeiro, no quadro clínico e evolução de peritonites em pacientes em diálise peritoneal, ocorridas na Unidade de Diálise do Hospital das Clínicas da Faculdade de Medicina de Botucatu nos últimos 17 anos. Entre 1994 e 2011, 192 episódios, que ocorreram em 118 pacientes, foram analisados. Os dados clínicos foram obtidos dos prontuários e fichas médicas específicas. Nas amostras de Staphylococcus foram realizados testes específicos fenotípicos e/ou genotípicos, para detecção de fatores de patogenicidade e de susceptibilidade à oxacilina e vancomicina; o perfil clonal foi determinado pela técnica de pulsed-field gel electrophoresis. Modelo de regressão logística foi utilizado para avaliar a influência dos fatores clínicos e microbiológicos no desfecho das peritonites. Além de S. aureus, outras espécies de Staphylococcus foram identificadas (S. epidermidis, S. haemolyticus, S. warneri, S. hominis, S. capitis, S. cohnii, S. saprophyticus, S. lugdunensis, S. simulans e S. xylosus). As peritonites por S. aureus apresentaram maiores taxas de ocorrência hipotensão... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Bacterial peritonitis remains as the main cause of technique failure of peritoneal dialysis (PD) and Gram-positive cocci, namely Staphylococcus spp., are the main etiological agents of such infections. Peritonitis caused by Staphylococcus aureus presents poorer prognostic than infections due to coagulase negative staphylococci (CoNS), even the antimicrobial resistance is more pronounced in the last agents. Host and bacterial factors may influence, therefore, peritonitis outcome. The objective of this study was to evaluate the influence of host and bacterial factors (clonal profile, virulence and antimicrobial resistance determinants in strains of staphylococci) on peritonitis clinical findings and outcome in patients from dialysis unit of Botucatu Medical School, Brazil, from 1994 until 2011. A hundred ninety-two episodes due to Staphylococcus were assessed. Genotypic or phenotypic tests were carried out to evaluate virulence factors and antimicrobial susceptibility for oxacillin and vancomycin; clonal profile was determined by pulsed-field gel electrophoresis. Logistic regression was used for the analysis of demographic, clinical, and microbiological factors influencing peritonitis outcome. Several species besides S. aureus were identified: S. epidermidis, S. haemolyticus, S. warneri, S. hominis, S. capitis, S. cohnii, S. saprophyticus, S. lugdunensis, S. simulans, and S. xylosus. Patients with S. aureus related peritonitis presented higher hypotension rate compared to the episodes due to CoNS (p<0.05). Oxacillin resistance was more frequent among CoNS (p<0.05) strains and vancomycin resistance was not detected, neither on S. aureus nor CoNS. S. aureus strains also presented higher enzymes and virulence genes than the others species (p<0.05), except... (Complete abstract click electronic access below) / Doutor

Thesis (M. Soc. Sc.)--University of Hong Kong, 2003.

No description available.

Thesis (Ph. D.)--Virginia Commonwealth University, 2007. / Prepared for: Dept. of Microbiology and Immunology. Bibliography: leaves 92-108. Also available online via the Internet.

Thesis (M. Soc. Sc.)--University of Hong Kong, 2003. / Also available in print.

Made available in DSpace on 2013-08-07T19:05:27Z (GMT). No. of bitstreams: 1 000386777-Texto+Completo-0.pdf: 1331431 bytes, checksum: 9edac4706a3d3ef38e1923efe97de1f0 (MD5) Previous issue date: 2006 / Objective: To observe the variations in serum glucose and insulin levels induced by the peritoneal exposition to glucose solution during a peritoneal equilibration test (PET), and to relate the findings with the solutes peritoneal transport rate. Patients and Method: A cross-sectional, observational, study using a modified PET procedure (4,25% glucose solution) that enrolled 34 prevalent peritoneal dialysis patients. Glucose and insulin serum levels were sequentially determined (at zero, 15, 30, 60, 120, 180, and 240 minutes) along the procedure, and an insulin resistance index (IR-HOMA) was computed. Categories of peritoneal transport were separated by quartiles of the dialisate/serum creatinine rate after 240 minutes of peritoneal exposition to the infused fluid (D4/PCr). Demographic and clinical variables were examined, and possible correlations among variables and categories of peritoneal transport were tested. Results: No difference for IR-HOMA, glucose and insulin levels was demonstrated among peritoneal transport categories. A direct correlation between the early glucose, and insulin peak increment and D4/PCr was evidenced. IR-HOMA was associated with the body mass index. Conclusion: Early glucose and insulin peak increment were associated with the solutes transport rate, as measured by the PET. The meaning of the findings on the outcome of patients with high peritoneal transport rate must be further evaluated. / Objetivo: Observar as variações de glicemia e insulinemia induzidas pela exposição da cavidade peritoneal à solução de glicose, durante teste de equilíbrio peritoneal (PET), e buscar relação com a velocidade de transporte peritoneal de pequenos solutos. Pacientes e Método: Estudo transversal, observacional, com 34 pacientes prevalentes em diálise peritoneal, submetidos a PET modificado (uso de glicose a 4,25%). Glicemia e insulinemia foram seqüencialmente determinadas sete vezes (em zero, 15, 30, 60 120, 180 e 240 minutos) ao longo do teste e índice de resistência a insulina (IR-HOMA) foi calculado. Categorias de transporte peritoneal foram definidas, na amostra, por quartís da razão dialisado/soro das concentrações de creatinina após 240 minutos de exposição do peritônio ao líquido (D4/PCr). Variáveis demográficas e clínicas foram computadas e possíveis correlações entre variáveis e categorias de transporte peritoneal foram testadas. Resultados: Não houve diferença para o IR-HOMA ou para medidas de glicemia e de insulinemia, entre as categorias de transporte peritoneal. Houve correlação direta entre os incrementos iniciais da glicemia, bem como a variação máxima de insulinemia e a variável D4/PCr – uma medida de velocidade de transporte de solutos pelo peritônio. O IR-HOMA relacionou-se diretamente com o índice de massa corporal. Conclusão: Os incrementos iniciais de glicemia e o pico máximo de insulinemia estão associados à velocidade de transporte peritoneal de pequenos solutos medida pelo PET. O significado destes achados sobre o prognóstico de pacientes com alto transporte deve ser mais bem avaliado.

Made available in DSpace on 2013-08-07T19:05:29Z (GMT). No. of bitstreams: 1 000326678-Texto+Completo-0.pdf: 874204 bytes, checksum: b8e45583ced3e993e571303b48afd0a2 (MD5) Previous issue date: 2004 / Objective: The aim of this study was to evaluate serum and dialysate nitric oxide (NO), and endothelial function in peritoneal dialysis patients in different peritoneal equilibration test (PET) categories. Methods: Cross-sectional study, in stable PD patients free of peritonitis for at least one month. Quartiles of the 4-h dialysate/plasma (D/P) ratio creatinine were used to classify the peritoneal membrane transport as low, low average, high average and high. NO metabolites were measured by chemiluminescence (n=41), and endothelial function was evaluated by arterial flow-mediated dilation (n=31). Results: Serum, 4-h and 24-h dialysate NO were not different among the PET categories. The 4-h dialysate/serum NO ratio (D/PNO) was also not different (p=0,096), but the data suggested the presence of a trend, increasing from low toward high peritoneal transport. Additionally, median (interquartile range) flowmediated dilation was not different between groups. There was correlation between 4-h dialysate and serum NO (r=0. 891, p<0. 001). D/PNO was negatively correlated with glucose transport (r=-0. 579, p<0. 001) and ultrafiltration (r=-0. 422, p<0. 001), and positively correlated with the D/P creatinine ratio (r=0. 533, p<0. 001). Conclusion: NO levels appear to reflect peritoneal permeability only. The dialysate NO level is not a sensible marker for local production of NO, in peritonitis-free patients. Peritoneal dialysis prescription is not dependent upon endothelial function. / Objetivo: O presente trabalho tem como objetivo avaliar os metabólitos do óxido níitrico (NO) no soro e no peritônio, e a função endotelial de pacientes em diálise peritoneal em diferentes categorias de transporte peritoneal, avaliados pelo teste de equilíbrio peritoneal. Método: Estudo transversal, em pacientes estáveis em DP. Quartis da razão dialisado/plasma (D/P) de creatinina, após 4 horas de permanência do líquido na cavidade, foram usados para classificar o transporte peritoneal em: baixo, baixomédio, alto-médio e alto. Os metabólitos do NO foram medidos por quimioluminescência (n=41), e a função endotelial avaliada através da dilatação mediada por fluxo da artéria braquial (n=31). Resultados: Os metabólitos do NO no soro, dialisado de 24 horas e de 4 horas não foram diferentes entre as categorias. A razão de NO no dialisado de 4 horas/soro (D/P de NO) também não foi diferente (p=0,096), mas houve tendência de aumento do baixo para o alto transportador. A mediana (intervalo interquartil) da dilatação mediada por fluxo não foi diferente entre os grupos. Houve correlação entre NO no soro e dialisado de 4 horas (r=0,891, p<0,001). A D/P de NO foi inversamente correlacionada com o transporte peritoneal de glicose (r=-0,579, p<0,001), e com a ultrafiltração (r=-0,422, p<0,001), e diretamente correlacionada com a D/P de creatinina (r=0,533, p<0,001). Conclusão: Os resultados sugerem que os níveis de NO refletem permeabilidade perioneal. Os metabólitos do NO não são um marcador da produção peritoneal de NO, em pacientes sem peritonite. A prescrição de diálise peritoneal não é dependente da função endotelial.

Objetivo: Avaliar a formação de aderências intraperitoneais em ratos após o implante peritoneal da tela de polipropileno comparada à tela de SIS, e o efeito do PAF como barreira anti-aderente à tela de polipropileno. Métodos: 55 ratos albinos foram randomizados em três grupos. O tipo de aderência, o percentual de tela coberta por aderência, e a força de rupturas das aderências foram avaliadas. Resultados: Os tipos de aderência 2 e 3 foram mais freqüentes no grupo 1 (polipropileno) e no grupo 3 (polipropileno+PAF); as do tipo 0 e 1 foram mais freqüentes no grupo 2 (SIS). A força media de ruptura foi de 1,58N (±0,719N) no grupo 1, 0,42N (±0,432N) no grupo 2 e 1,23N (±0,432N) no grupo3. Mais de 50% da tela estava coberta por aderências em 12 (80%) casos do grupo 1, em 4 (20%) casos do grupo 2 e em 16 (84,2%) casos do grupo 3. O grupo 2 foi significativamente diferente (p<0.001) dos demais grupos. Conclusão: O uso intraperitoneal da tela de polipropileno levou a elevados índices de aderência, e o uso de PAF como barreira anti-aderente não reduziu estes índices. O implante intraperitoneal de SIS revelou índices baixos de aderências peritoneais.

Made available in DSpace on 2014-06-11T19:31:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2002-08-14Bitstream added on 2014-06-13T20:47:25Z : No. of bitstreams: 1 peiro_jr_dr_jabo.pdf: 1256820 bytes, checksum: ef439522328684a352aaeec5533665d3 (MD5) / A endotoxemia é a maior causa de mortalidade em eqüinos. O desencadeamento da endotoxemia clínica nos eqüinos é freqüente em decorrência da elevada sensibilidade desta espécie aos lipopolissacarídeos (LPS) da membrana externa de bactérias Gram-negativas liberados para a circulação durante o processo de choque séptico. Em resposta a esta agressão, o organismo libera substâncias pró-inflamatórias, as citocinas, na corrente circulatória. Dentre estas, o fator de necrose tumoral (TNF) tem sido implicado como a principal citocina no desencadeamento do quadro endotoxêmico dos eqüinos, sendo encontrados níveis elevados desta citocina, tanto séricos como peritoneais, em eqüinos com cólica. Os anestésicos locais, com o grupamento amídico em sua estrutura molecular, têm exibido ação inibitória sobre a resposta inflamatória por bloqueio da resposta funcional da atividade dos polimorfonucleares na liberação das citocinas. A lidocaína tem sido utilizada com sucesso, como anestésico local, na prevenção da vasoconstrição mesentérica durante o choque endotóxico em cães, assegurando proteção a esta agressão e demonstrando ser efetiva no tratamento de processos inflamatórios localizados no intestino delgado quando aplicada topicamente na serosa, ou na forma de bolus seguida de infusão contínua para o tratamento do ileus adinâmico. O principal objetivo deste ensaio foi estudar os efeitos da infusão contínua de lidocaína, sobre a resposta clínica, laboratorial e na resposta inflamatória, avaliados através da produção do TNF alfa, antes e após a injeção intraperitoneal de LPS em eqüinos, e avaliar a segurança desta dose de lidocaína neste modelo experimental. Com base nos achados clínicos e laboratoriais deste estudo, concluiu-se que a utilização da lidocaína após a administração do LPS foi mais efetiva na inibição da produção... / Endotoxemia is the major cause of mortality in horses. The onset of signs of endotoxemia in horses occurs frequently due to their sensitivity to the release of lipopolysaccharides (LPS) of the outer membranes of gram negative bacteria during the septic shock. In response, the host releases proinflammatory cytokines into the bloodstream. Tumor necrosis factor-a is an important cytokine released during the onset of endotoxemia and increased levels of TNF can be found in peritoneal fluid as in serum of colicky horses. Amide local anaesthetics have shown to inhibit polymorphonuclear release of cytokines. Lidocaine has also been shown to be an effective method for treating small intestine inflammatory conditions when injected as an intravenous bolus followed by continuous infusion or acting as a prokinetic drug during postoperative ileus. The main goal of this study was to evaluate the effects of lidocaine continuous infusion on clinical, laboratory responses and inflammatory response, in terms of TNF-alpha activity, before and after intraperitoneal injection of lypopolisaccharides in horses. Based on the clinical and laboratory findings in this study, we concluded that lidocaine administered after LPS was more effective in inhibiting TNF-alpha activity, although it was not able to avoid the increasing of inflammatory cells in the peritoneal fluid.