Orientador: Rogério Saad Hossne / Resumo: Introdução: As Doenças inflamatórias intestinais (DII), que tem como principais entidades a Retocolite Ulcerativa (RCU) e a Doença de Crohn (DC), tem altas taxas de incidência e prevalência em países desenvolvidos, especialmente da Europa e América do Norte, porém com aumento progressivo de sua frequência em todas os continentes. Este estudo visa estimar as taxas de incidência e prevalência das DII no Estado de São Paulo, Brasil, entre os anos de 2012 e 2015, e correlacionar os resultados com dados nacionais sobre estas doenças. Material e Método: Este é um estudo observacional analítico, do tipo descritivo e transversal. Foram incluídos dados epidemiológicos de 22.638 pacientes que iniciaram seu tratamento para Doença Inflamatória Intestinal através do programa de fornecimento gratuito de medicamentos do Estado de São Paulo, entre os anos de 2012 e 2015. As variáveis analisadas foram a data do início do tratamento, o diagnóstico clínico (DC ou RCU), a idade, gênero, cor/raça/etnia dos pacientes, assim como sua região de residência no Estado de São Paulo. As análises estatísticas incluíram média e desvio padrão para variáveis quantitativas. O nível de significância adotado foi de 1% Resultados: A taxa de incidência de DII no Estado de São Paulo foi, em média, de 13,31 casos novos / 100.000 habitantes / ano, enquanto a prevalência de DII no Estado de São Paulo foi de 52,5 casos / 100.000 habitantes. Os portadores de DC somavam 10.451 (46,16%), e os de RCU somavam 12.187 (53,83... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Inflammatory bowel disease (IBD), which has as its main entities Ulcerative Colitis (UC) and Crohn's Disease (CD), have high rates of incidence and 11 prevalence in developed countries, especially in Europe and North America, but with increasing frequency in all continents. This study aims to verify the incidence and prevalence rates of IBD in São Paulo State, Brazil, between the years 2012 and 2015, and correlate with the national data on these diseases. Casuistic and Methods: This is an observational, descriptive and cross-sectional study. We included data from 22.638 patients who started their treatment for Inflammatory Bowel Disease through the Program of free medication supply of São Paulo State, between the years of 2012 and 2015. The variables analyzed were the date of beginning of treatment with drugs provided by the clinical diagnosis (CD or UC), the age, gender, color/race/ethnicity of the patients, as well as their region of residence in São Paulo State. Statistical analyses included mean and standard deviations for quantitative variables. The level of significance adopted was 1% Results: The incidence rate of IBD in the State of São Paulo was 13.31 new cases / 100.000 inhabitants per year, while the overall prevalence of IBD in the state of São Paulo was 52,5 cases/100.000 inhabitants. The patients with CD were 10,451 (46.16%), and those with UC were 12,187 (53.83%), from 1 to 97 years of age, with a mean of 45.5 years (SD = 16.7), of wich 9,124 (40.30%) were male... (Complete abstract click electronic access below) / Doutor

INTRODUCTION: The management of pediatric patients with ulcerative colitis (UC) is dependent upon the ability to detect meaningful changes in disease status. This is currently done using validated patient-reported clinical disease activity indices, including the Pediatric Ulcerative Colitis Activity Index (PUCAI). While useful for global assessments completed during ambulatory office visits, the sensitivity of this metric may be insufficient to reflect more subtle changes in disease activity or response to medical therapy in hospitalized patients. Intravenous steroids are typically employed in the management of patients admitted for acute exacerbations of UC symptoms. These are typically manifest by worsening bloody diarrhea, abdominal pain, and worsening anemia. There is presently no way of predicting whether a patient admitted for UC will respond to steroid therapy. Current paradigms dictate a five-day trial before considering a transition to more potent medical or definitive surgical approaches to the management of refractory colitis. The development of more sensitive and reliable biomarkers or disease activity metrics could enable clinicians to more expediently identify steroid non-responders. This would minimize patient morbidity, decrease risk of complication, and lower overall cost of care. Previous studies have demonstrated that changes in fecal lactoferrin (FLA) correlate with disease activity in patients with UC. OBJECTIVES: To analyze the predictive value of FLA in the response to steroid treatment of patients admitted for management of UC. METHODS: We recruited pediatric inpatients with UC in the Division of Gastroenterology, Hepatology and Nutrition at Boston Children’s Hospital who were hospitalized for treatment of a flare of their UC symptoms. After obtaining patient consent, we collected a stool sample on days 1 and 3 of their hospital stay. We sent samples to TECHLAB® Inc. (Blacksburg, VA) to be analyzed for levels of FLA. We compared Day 1, Day 3 and ∆FLA (Day 1 – Day 3) in two patient groups: those that responded to conventional steroid therapy and those that required rescue medical or surgical therapy. We reported statistical significance with the Wilcoxon signed-rank test. RESULTS: Of 67 patients consented for the study, 30 provided stool samples on both days 1 and 3 of their inpatient hospitalization. Of the 30 patients, 63.3% responded to steroids while 36.7% required rescue therapy with immunomodulators. ∆FLA for responders, 43.6μg/mL(-239.0, 331.6) (median(interquartile range)), did not differ significantly from non-responders, -74.1μg/mL(-296.7, 221.7), P = 0.3. CONCLUSIONS: Our findings do not demonstrate that measurement of changes in quantitative FLA over three days can be used to assess acute responses to steroid therapy. Increasing the sample size may allow us to better delineate subtle differences between responders and non-responders to steroid therapy.

Introduction Inflammatory bowel disease (IBD) is a chronic gastrointestinal disorder that encompasses two major subtypes; Crohn’s Disease (CD) and Ulcerative Colitis (UC). Our knowledge regarding disease pathogesis is rapidly increasing. However, these disease entities provide challenges in diagnosis, monitoring of disease activity and assessing individual response to treatment, because there is a lack of validated clinical biomarkers. Metabolomics involves the study of numerous analytes that have very diverse physical and chemical properties and occur in a wide concentration range. Early evidence suggests there is potential for metabolomic profiling to be used in the differentiation of CD and UC. However, knowledge is limited regarding the metabolic changes seen in relation to disease activity or to medical or surgical treatments. Aims A metabolomics approach was taken to determine whether metabolomic profiles could distinguish between patients with CD or UC and healthy controls. We also aimed to define the relationship between metabolomic profile and disease activity, and to determine the effect of medical (anti-TNFa agents) and surgical treatment on the metabolome. Methods A metabolomics approach was undertaken. Serum and urine sample sets were collected from a total of 41 patients with ulcerative colitis, 43 patients with Crohn’s disease, and 62 healthy controls (HC). In order to allow a comparison of metablomic profile and disease activity, 4 sample sets were taken from the same patient at 3 monthly intervals over the period of one year. Those patients undergoing either surgical or biological treatment had sample sets taken pre and post intervention. Metabolomic analysis using gas chromatography time of flight mass spectrometry (GC-ToF-MS) and ultra-high performance liquid chromatography Fourier Transform mass spectrometry (UHPLC-FTMS) was carried out on both serum and urine. Results Serum and urine GC-ToF-MS and UHPLC-FTMS metabolomic analyses show differentiation between UC, CD and healthy controls, most significantly in urine analyses. No significant differentiation was seen in pre- and post-surgical patients, or pre- and post-biological therapy patients. It was possible to differentiate surgical patients from healthy controls, especially in the urine analyses. Metabolite identification revealed consistently more dietary variation in the healthy controls than in the IBD patients. Significant differences (p < 0.05) were seen between healthy controls and IBD patients in classes of metabolites relating to the citric acid cycle and the uronic acid pathway, as well as amino acids, fatty acids and cholesterols. The behaviour or location of disease, or the disease activity score did not appear to influence the metabolome in either serum or urine analyses using GC-ToF-MS and UHPLC-FTMS. Conclusion Metabolomic profiling of urine and serum in IBD may provide a novel methodology aiding both clinical diagnosis through biomarker development, and advancing knowledge of disease pathogenesis.

As doenças inflamatórias intestinais (DII) são doenças crônicas incapacitantes com significativa morbidade. A Doença de Crohn (DC) e a Retocolite Ulcerativa (RCU) são as maiores representantes. O diagnóstico é baseado na suspeita clínica e complementado por achados endoscópicos, histopatológicos e radiológicos. Como não existe um teste padrão, o diagnóstico destas doenças permanece sendo um problema para gastroenterologistas e patologistas. O desenvolvimento de terapias mais específicas torna cada vez mais importante o diagnóstico preciso para uma escolha terapêutica individualizada. A imunidade inata está ativada de forma semelhante nas duas DII, porém na RCU os linfócitos CD4 TH2 são os principais envolvidos. Estes linfócitos expressam o receptor CD30 na sua membrana plasmática e produzem citocinas que ativam os eosinófilos. Levando em consideração o conhecimento fisiopatológico atual, este estudo tem como principal objetivo avaliar a relevância da expressão do marcador CD30 por imuno-histoquímica, a contagem de eosinófilos e as características histopatológicas no diagnóstico diferencial das DII. Foram avaliados 185 pacientes de um ambulatório especializado em um Hospital Universitário (105 com DC 80 com RCU). O diagnóstico foi estabelecido pelos critérios de Lennard-Jones, revisados por um gastroenterologista especializado e corroborado por cinco anos de seguimento. As biópsias foram realizadas por diversos profissionais na rotina assistencial, porém todas foram analisadas por um patologista com experiência em tubo digestivo. Nenhum paciente estava usando tratamento no momento da coleta das biópsias. As variáveis histopatológicas que demonstraram poder estatístico para auxiliar no diagnóstico diferencial foram a extensão do processo inflamatório para a submucosa, a presença de granuloma, erosões aftóides e variabilidade de acometimento entre os fragmentos. Avaliando estas características em conjunto, foi possível obter uma acurácia de 69.1% para o diagnóstico diferencial entre DC e RCU. Estudando o segmento mais alterado encontrou-se uma mediana de eosinófilos de 42 (25,5 – 63,5), nos pacientes com DC e 107 (67 – 123) nos pacientes com RCU (p< 0.001). Assumindo como ponto de corte um número V a 70 eosinófilos, a sensibilidade foi de 78,3% e a especificidade de 71% favorecendo o diagnóstico de RCU, a área sob a curva ROC foi de 0,767 (IC 95%: 0,696–0,838). A imuno-histoquímica com CD30 demonstrou uma mediana de 3 células CD30+ (2-6) na DC e 33 (24-52) na RCU, demonstrando uma diferença estatística significativa para o diagnóstico diferencial entre as duas doenças (p<0,001). Além disso, nos pacientes com RCU as células CD30+ estavam distribuídas mais frequentemente em agrupamentos nos centros dos folículos linfoides. O ponto de corte determinado pela curva ROC foi de 15 células marcadas (S = 97,5%, E = 94,3%, RV + = 17,1; RV = 0.03, área sob a curva: 0.967, IC 95%: 0.941 - 0.993). Todos estes parâmetros estudados tiveram capacidade discriminatória para o diagnóstico de DC e RCU. Designando um valor para cada variável, baseado no poder estatístico de cada uma e de forma a obter uma soma de 10 pontos no total, foi construído um escore histopatológico para o diagnóstico da DC. Considerando-se a soma das variáveis V 5 obteve-se uma especificidade de 100% e uma sensibilidade de 86,8%. Considerando 4 como ponto de corte do escore, a sensibilidade aumenta para 95,3%, e a especificidade reduz de 100% para 94,9%. Concluindo, o uso rotineiro da avaliação das características histopatológicas descritas em associação com a contagem de eosinófilos e células CD30+ proporciona uma alta acurácia no diagnóstico diferencial entre DC e RCU. Todos os parâmetros estudados são de fácil avaliação tanto por patologistas especialistas quanto generalistas. Sugere-se a realização de um estudo prospectivo para validação deste escore. / Inflammatory bowel diseases (IBD) are chronic disabling diseases with significant morbidity, being Crohn’s disease (DC) and Ulcerative colitis (UC) their greatest representatives. Diagnosis is based on clinical suspicious and complemented by endoscopic, histopathological and radiological findings. However, there is not a gold standard test, so it remained a problem for gastroenterologists and pathologists. The development of more targeted therapies makes it even more important to establish an accurate diagnosis for a better individualized therapeutic approach. Innate immune response is activated in both IBD, but in UC the lymphocytes CD4 TH2-like are mainly engaged, this kind of lymphocytes has CD30 expressed in their plasma membrane and produces cytokines that activate eosinophils. Considering the current pathophysiological knowledge, this study aimed to evaluate the relevance of CD30 expression by immunohistochemical, eosinophil count and histopathological features in differential diagnosis of IBD. A total of 185 patients were evaluated (105 CD/ 80 UC). Patients were followed at a specialized clinic of a university hospital, diagnosed according to Lennard- Jones criteria reviewed by a gastroenterologist expert and corroborated by five years of follow up. Biopsy samples were taken by different professionals in routine care, but analyzed by an experienced gastrointestinal pathologist. None of the patients were using treatment at the time of biopsy. Of all the pathological variables assessed, those that had statistical capacity to assist in the differential diagnosis were extension of the inflammatory process to submucosa, granuloma, aphthous erosion and variability of involvement between fragments. Evaluating these variables together, an accuracy of 69.1% in the differential diagnosis between CD and UC was found. Assessing the most altered sample the median of eosinophils was 42 (25.5 – 63.5) in CD patients and 107 (67 – 123) in UC patients (p< 0.001). Assuming a cutoff V 70 eosinophils, the sensitivity was 78,3% and specificity of 71% favoring the UC diagnosis, the area under the ROC curve was 0.767 (CI 95%: 0,696–0,838). Immunohistochemical CD30+ cells presented with a median of 3 cells (2-6) in CD and 33 cells (24-52) in UC, demonstrating a highly significant statistical difference between the two diseases (p<0.001). Besides, CD30+ cells were distributed most clustered in the center of lymphoid follicles in UC patients. The cutoff determined by ROC curve was 15 (S = 97.5%, E = 94.3%, LR + = 17.1;-RV = 0.03, AUC: 0.967, 95% CI: 0.941 - 0.993). All these parameters studied had discriminatory capacity for diagnosis of CD and UC. A value was assigned to each variable based on the statistical power of each, making a total sum of 10 points to build a histopathological score for the CD diagnosis. Setting the cutoff as V 5 we found a specificity of 100% and a sensitivity of 86.8%. When we consider as cutoff as V 4 points the score sensitivity comes to 95.3%, and reduces the specificity of 100% to 94.9%. In conclusion, the use of routine assessment of the histopathological features described previously in association with the eosinophils and CD30+ cells count provides a high accuracy for CD and UC differential diagnosis. All parameters assessed here are easily performed by pathologists specialists and generalists. The next step seems to be the validation of this score in a prospective study.

Introdução: A retocolite ulcerativa indeterminada é uma doença inflamatória que envolve exclusivamente o cólon e o reto, sendo caracterizada por infiltrado leucocitário e úlceras superficiais na mucosa intestinal. A produção e liberação de espécies reativas de oxigênio e nitrogênio parecem ser cruciais na determinação da fisiopatologia da doença, pois resultam em dano oxidativo. A partir dessas informações, a busca por opções terapêuticas com propriedades antioxidantes são importantes e têm sido testadas na colite experimental. Objetivo: Este estudo tem como objetivo avaliar os efeitos do extrato seco da planta Boswellia serrata em modelo experimental de colite induzida por ácido acético sobre os danos teciduais, a pressão anal esfincteriana, o estresse oxidativo, a atividade das enzimas antioxidantes superóxido dismutase (SOD) e glutationa peroxidase (GPx) e atividade da glutationa (GSH), a concentração dos metabólitos do óxido nítrico e expressão da enzima óxido nítrico sintase induzível (iNOS) por imunohistoquímica. Material e métodos: Foram utilizados 25 ratos Wistar machos, com peso médio de 350g, divididos em 5 grupos: Controle (CO); Controle+Boswellia serrata (C+B); Colite (CL); Colite+Boswellia serrata (CL+B) e Boswellia serrata+Colite (B+CL). Os animais foram submetidos à administração intracolônica por enema com solução de ácido acético diluído a 4% e com volume de 4 mL. O tratamento com o extrato aquoso da planta via oral, na dose de 34,2 mg/Kg diluído em 4 mL de solução fisiológica, ocorreu uma vez ao dia durante 48 horas antes e após a indução da colite. Foi realizada a medida de pressão anal esfincteriana dos animais. As análises histológicas do intestino foram através da coloração de Hematoxilina-Eosina e realizada imunohistoquímca com anticorpo iNOS. O homogeneizado do intestino foi utilizado para avaliação da lipoperoxidação (LPO) através das substâncias reativas ao acido tiobarbitúrico (TBARS), avaliação dos metabólitos do óxido nítrico pela técnica de nitritos e nitratos totais, avaliação da atividade das enzimas antioxidantes SOD e GPx e avaliação da GSH. Resultados: Na análise da pressão anal esfincteriana os animais dos grupos CL+B e B+CL apresentaram um aumento significativo em relação ao grupo CL. Nos níveis de LPO e metabólitos do óxido nítrico foi observada uma diminuição significativa nos grupos CL+B e B+CL quando comparados ao grupo CL. A atividade da SOD mostrou um aumento no grupo CL e uma diminuição significativa nos grupos CL+B e B+CL equivalendo à média do grupo CO. A GPx e GSH apresentaram um aumento significativo nos grupos CL+B e B+CL em relação ao grupo CL. Conclusão: Sugerimos que a administração do extrato da planta Boswellia serrata possa ser uma possibilidade de terapia antioxidante na colite ulcerativa. / Introduction: Ulcerative rectocolitis is an inflammatory disease that involves only the colon and rectum, being characterized by leukocyte infiltrate and superficial ulcers in the intestinal mucosa. The production and release of reactive oxygen and nitrogen species appears to be crucial in determining the pathophysiology of the disease, since both result in oxidative damage. Therefore, the search for treatment options with antioxidant properties is important currently and has been tested in experimental colitis. Objective: This study aimed to evaluate the effects of dry extract of Boswellia serrata plant in an experimental model of colitis induced by acetic acid on tissue injury, anal sphincter pressure, oxidative stress, on the activity of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione (GSH), concentration of nitric oxide metabolites and expression of inducible nitric oxide synthase enzyme (iNOS) by immunohistochemistry. Methods: We used 25 male Wistar rats with an average weight of 350g, divided into 5 groups: control (CO), Control + Boswellia serrata (CO+B); Colitis (CL); Colitis + Boswellia serrata (CL+B) and Boswellia serrata + colitis (B+CL). The animals were submitted to intracolonic administration by enema with acetic acid solution diluted to 4% in a volume of 4 ml. The treatment with aqueous plant extract was performed orally at a dose of 34.2 mg/kg diluted in 4 ml of saline. The administration occurred once daily for 48 hours before and after the induction of colitis. We performed the measurement of anal sphincter pressure animals. Histological analyzes of bowel the after were made staining with hematoxylin-eosin and immunohistochemistry, performed with iNOS antibody. The homogenized intestine was used for evaluation of lipid peroxidation (LPO) through thiobarbituric acid reactive substances (TBARS), assessment of nitric oxide metabolites by the technique of total nitrites and nitrates and evaluation of the antioxidant enzymes SOD, GPx and GSH. Results: The analysis of anal sphincter pressure of the animals in groups CL+B and B+CL showed a significant increase when compared to the CL group. LPO and nitric oxide metabolites levels demonstrated significant decrease in groups CL+B and B+CL when compared to CL. SOD activity showed an increase in CL group and a significant decrease in groups CL+B and B+CL, remainig to the average of the CO group. The GPx and GSH showed a significant increase in groups CL+B and B+CL group when compared to CL group. Conclusion: We suggest that the administration of Boswellia serrata plant extract may be a possibility of antioxidant therapy in ulcerative colitis.

O nosso objetivo foi mensurar os níveis de Interleucina-6 (IL-6) no fluido gengival de pacientes com periodontite e doença inflamatória intestinal (DII), comparando-os com pacientes sistemicamente saudáveis, com periodontite. Como objetivo secundário será avaliada a IL-6 no soro desses pacientes. Foram selecionados 15 pacientes com doença de Crohn (DC, idade média 38.2, DP 11.4 anos), 15 com retrocolite ulcerativa idiopática (RCUI, 45.0 10.5 anos) e 15 pacientes saudáveis (C, 42.1 7.8 anos). A Profundidade de bolsa (PB), nível de inserção clínica (NI), presença de placa e de sangramento a sondagem foram avaliados em seis sítios por dente. O fluido gengival foi coletado de quatro sítios com periodontite (PP: PB &#8805; 5mm, NI &#8805; 3mm) e quatro sítios com gengivite (GP: PB &#8804; 3mm e NI&#8804; 1mm), em dentes diferentes, com pontas de papel absorvente pré-fabricadas. O soro destes pacientes também foi coletado. A análise da IL-6 foi realizada pelo LUMINEX. A quantidade total e concentração da IL-6 estavam significantemente maiores no fluido gengival dos sítios PP do grupo RCUI quando comparados aos sítios PP do grupo controle (p=0.028; p=0.044, respectivamente). O grupo DC apresentou a quantidade total de IL-6 significantemente maior no sítio PP do que no GP (p=0.028). Já no soro, a IL-6 não diferiu entre os grupos. Sendo assim, pode-se concluir que os indivíduos com retrocolite ulcerativa idiopática apresentavam níveis mais altos de IL-6 nos sítios com periodontite, o que pode indicar um importante papel dessa citocina no estabelecimento e progressão da doença periodontal nesses pacientes. / Our aim was to measure the levels of interleukin-6 in gingival crevicular fluid (GCF) from patients with periodontitis and inflammatory bowel disease (IBD), and compare with systemically healthy controls with periodontitis. Besides, was measured IL-6 level in serum. Fifteen patients with Crohns disease (CD, mean age 38.2 11.4 years), 15 with ulcerative colitis (RCUI, 45.0 10.5) and 15 controls (C, 42.1 7.8) participated in this study. Probing pocket depth (PPD), attachment loss (CAL), presence of plaque and presence of bleeding on probing, were assessed in six sites per tooth. In each subject, GCF from four sites with gingivitis (GP- PPD&#8804; 3mm and CAL &#8804; 1mm) and from 4 sites with periodontitis (PP- PPD &#8805; 5mm and CAL &#8805; 3mm), on different teeth, were collected with filter strips. The serum of those patients was also collected. The IL-6 was analyzed in the Luminex. The total amount and the concentration of IL-6 in GCF was significantly higher in PP sites from RCUI than in controls (p=0.028 e p=0.044, respectively). The total amount of IL-6 was significantly higher in PP than GP sites in the DC group (p=0.028). In serum, IL-6 does not differ between groups. Therefore was concluded that subjects with ulcerative colitis showed higher levels of IL-6 in periodontitis sites, which might indicate an important role of this cytokine in the onset and progression of periodontal disease in such patients.

Colonic stromal cells provide critical structural support but also regulate immunity, tolerance and inflammatory responses in the mucosa. Substantial variability and plasticity of mucosal stromal cells has been reported but a paucity of distinct marker genes exist to identify distinct cell states. Here single-cell RNA-sequencing is used to document heterogeneity and subtype specific markers of individual colonic stromal cells in human and mouse. Marker-free transcriptional clustering of fibroblast-like cells derived from healthy human tissue reveals distinct populations corresponding to myofibroblasts and three transcriptionally and functionally dissimilar populations of fibroblasts. A SOX6 high fibroblast subset occupies a position adjacent to the epithelial basement membrane and expresses multiple epithelial morphogens including WNT5A and BMP2. Additional fibroblast subtypes show specific enrichment for chemokine signalling and prostaglandin E₂ synthesis respectively. In ulcerative colitis, substantial remodelling occurs with depletion of the SOX6 high population and emergence of an immune enriched population expressing genes associated with fibroblastic reticular cells including CCL19, CCL21 and IL33. A large murine dataset comprising over 7,000 colonic mesenchymal cells from an acute colitis model and matched healthy controls reveals strong preservation of the SOX6 high and myofibroblast transcriptional signatures. Unsupervised pseudotemporal ordering is used to relate fibroblast subsets to one another producing a branched developmental hierarchy that includes a potential progenitor population with mesothelial characteristics at its origin. This work provides a molecular basis for re-classification of colonic stromal cells and identifies pathological changes in these cells underpinning inflammation in UC.

Submitted by THAISY MILANELLI CANHIZARES null (thaisymilanelli@yahoo.com.br) on 2017-08-24T01:15:33Z No. of bitstreams: 1 DEFESA final.pdf: 700718 bytes, checksum: 9c3cbfade3105f6f93da40a95c86b3c6 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-08-25T14:14:51Z (GMT) No. of bitstreams: 1 canhizares_tm_me_bot.pdf: 700718 bytes, checksum: 9c3cbfade3105f6f93da40a95c86b3c6 (MD5) / Made available in DSpace on 2017-08-25T14:14:51Z (GMT). No. of bitstreams: 1 canhizares_tm_me_bot.pdf: 700718 bytes, checksum: 9c3cbfade3105f6f93da40a95c86b3c6 (MD5) Previous issue date: 2017-08-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Retocolite Ulcerativa (RU) é um tipo de patologia que acomete o cólon intestinal, se apresentando na forma de lesões superficiais de gravidade variável. Não possui causa definida, mas sabe-se que é influenciada por fatores genéticos e ambientais, na qual, esse último, inclui um desequilíbrio na composição de espécies da microbiota intestinal. Escherichia coli (E. coli), uma das bactérias que se encontra aumentada nesses pacientes, tem sido foco de estudos de caracterização, com o objetivo de esclarecer sua participação na etiologia ou complicação dos sintomas da doença. Esse trabalho adotou essa abordagem para a caracterização de uma coleção de E. coli isoladas de portadores de RU atendidos no Hospital das Clínicas da Universidade Estadual Paulista Júlio de Mesquita Filho (HC/UNESP) de Botucatu, com base em sua capacidade de produção de biofilme, sorotipagem e filotipagem. Juntamente a esses testes, foi realizada uma revisão bibliográfica sobre a possível relação da E. coli com a RU. O objeto de estudo dos testes foi uma coleção de E. coli composta por 68 isolados bacterianos de 34 portadores de RU e 44 de 22 indivíduos controle (CO). A tipagem bacteriana teve como foco genes que identificam os sorogrupos O25 e O83 e determinação de filogrupos da coleção de referência de E. coli (EcoR – A, B1, B2 e D). Os resultados obtidos foram: 1) predomínio de E. coli dos filogrupos B2 e A nos grupos CO (54,5% x 26,5%, p=0,01) e de portadores de RU (32,4% x 9,1%, p=0,04) respectivamente, 2) no grupo portador de RU, 8,8% e 11,8% dos indivíduos apresentaram os sorogrupos O25 e O83, respectivamente e, entre os CO, a prevalência de ambos os sorogrupos foi de 4,5% e, 3) isolados produtores de biofilme forte (Fo), moderado (Mo) e fraco (Fra) foram encontrados em 45,5%, 22,7% e 27,3% dos CO, respectivamente. Em portadores de RU, a prevalência foi de 32,4%, 8,8% e 14,7%, respectivamente. A divergência nos dados de filotipagem em relação à literatura denota o caráter de extensa variabilidade observada nas populações naturais de E. coli e que dificulta sua vinculação com a causa da RU. A ausência de diferença na prevalência de isolados produtores de biofilme entre os grupos sugere que tal propriedade não pode ser vinculada a um eventual potencial de E. coli em provocar ou complicar os sintomas da RU. A análise bibliográfica mostrou resultados divergentes sobre a relação da E. coli na RU, possivelmente devido às variações no método de colheita, características teciduais e método de quantificação das culturas, sendo necessário mais pesquisas sobre o tema para sua maior clareza. / Ulcerative colitis (UC) is a type of pathology that affects the intestinal colon, presenting as superficial lesions of different severity. It has no defined cause, but it is known to be influenced by genetic and environmental factors, which includes an imbalance in the composition of species of the intestinal microbiota. Escherichia coli (E. coli), one of the bacteria that is increased in these patients, has been the focus of characterization studies, to clarify its participation in the etiology or complication of the disease’s symptoms. Following a line of research already consolidated in our laboratory, this work adopted this approach for the characterization of a collection of E. coli isolated from UC patients treated at the HC / UNESP of Botucatu, based on its biofilm production capacity, serotyping and filotyping. Also, a literature review was performed on the possible relationship between E. coli and UC. The study’s object of these tests was a collection of E. coli composed of 68 bacterial isolates from 34 UC carriers and 44 from 22 control individuals (CO). Bacterial typing focused on genes that identify the O25 and O83 serogroups and determination of phylogroups from the E. coli reference collection (EcoR - A, B1, B2 and D). The results obtained were: 1) Predominance of E. coli of the phylogenetic groups B2 and A in the CO groups (54.5% x 26.5%, p = 0.01) and in the UC group (32.4% x 9, 1, p = 0.04), respectively. 2) In the UC group, 8.8% and 11.8% of the individuals had serogroups O25 and O83, respectively, and among CO, the prevalence of both serogroups was 4,5% and, 3) Isolated producers of strong (St), moderate (Mo) and weak (We) biofilms were found in 45,5%, 22,7% e 27,3% of the CO, respectively. In UC patients, the prevalence was 32.4%, 8.8% and 14.7%, respectively. The divergence in the data of phylotyping in relation to the literature denotes the character of extensive variability observed in the natural populations of E. coli and that makes it difficult to be linked to the cause of the UC. The absence of a difference in the prevalence of biofilm isolates among the groups suggests that such property can’t be linked to an eventual potential of E. coli to cause or complicate UC symptoms. The literature analysis showed divergent results on the relationship of E. coli and UC, possibly due to variations in the collection method, tissue characteristics and quantification method of the cultures, being necessary more researches on the subject for its greater clarity.

Submitted by Élen Farinelli de Campos Silva (elenfarinelli@hotmail.com) on 2018-04-11T01:53:32Z No. of bitstreams: 1 Dissertacao.pdf: 1220781 bytes, checksum: 569fcdc5b7a92337431b3ae556c60152 (MD5) / Rejected by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: problema 1: ficha catalográfica Falta ficha catalográfica. A ficha deve ser inserida no arquivo PDF logo após a folha de rosto do seu trabalho. Assim que tiver efetuado a correção ou correções submeta o arquivo em PDF novamente. 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Agradecemos a compreensão. on 2018-04-17T12:09:50Z (GMT) / Submitted by Élen Farinelli de Campos Silva (elenfarinelli@hotmail.com) on 2018-04-18T13:24:35Z No. of bitstreams: 1 Tese Defesa Elen 18 abril 2018 - Ellen.pdf: 1319266 bytes, checksum: 69a7a76892b1bad189a29b29c5bd54eb (MD5) / Approved for entry into archive by Luciana Pizzani null (luciana@btu.unesp.br) on 2018-04-19T12:52:32Z (GMT) No. of bitstreams: 1 silva_efc_me_bot.pdf: 1319266 bytes, checksum: 69a7a76892b1bad189a29b29c5bd54eb (MD5) / Made available in DSpace on 2018-04-19T12:52:32Z (GMT). No. of bitstreams: 1 silva_efc_me_bot.pdf: 1319266 bytes, checksum: 69a7a76892b1bad189a29b29c5bd54eb (MD5) Previous issue date: 2018-02-27 / Introdução: as Doenças Inflamatórias Intestinais (IBD), representadas pela Doença de Crohn e Retocolite Ulcerativa (RCU), podem evoluir com sintomas incapacitantes que comprometem a qualidade de vida de seus portadores. A identificação precoce de doença grave permite terapêutica inicial mais agressiva com menores taxas de complicações e hospitalizações, cirurgias e morte. O objetivo do presente estudo foi identificar as variáveis associadas à necessidade de hospitalização, cirurgia de colectomia, evolução para câncer colorretal e óbito em pacientes portadores de RCU. Metodologia: foi realizado estudo observacional e retrospectivo com coleta de dados de pacientes acompanhados no Ambulatório de DII da Faculdade de Medicina de Botucatu, totalizando 284 pacientes elegíveis. Excluímos 30 pacientes com dados faltantes, totalizando 254 pacientes analisados. As características demográficas, tabagismo, aspectos clínicos como extensão e atividade da doença, além da presença de manifestações extraintestinais (MEI), medicamentos em uso e comorbidades foram avaliados. Os defechos considerados foram necessidade de hospitalização por complicações da doença, necessidade de colectomia, evolução para câncer colorretal ou óbito. Análise estatística: análise descritiva e testes de associação. Foram realizadas análises de regressão logística univariada e multivariada para avaliar as variáveis associadas ao desfecho. As variáveis de desfecho foram necessidade de hospitalização, colectomia, câncer colorretal e óbito. A curva de sobrevida foi realizada utilizando o teste Log Rank, no qual o evento inicial foi a data do diagnóstico e o evento final foi a necessidade de hospitalização, colectomia, câncer colorretal ou óbito ou o último contato com o paciente. Nível de significância p <0,05. Resultados: a média de idade foi de 46,64 (± 16,88) anos, 62,99% eram mulheres, 49,61% apresentavam pancolite e 45,68% estavam em remissão clínica. Em relação ao tabagismo, 66,40% dos pacientes eram não-fumantes, 28,06% ex-fumantes e 5,53% fumantes. MEI foi observada em 52,36% dos pacientes e 10,63% dos pacientes estavam em uso de terapia biológica. Noventa e três pacientes (29,06%) necessitaram de hospitalização. As variáveis associadas com hospitalização foram extensão pancolite, presença de colangite esclerosante primária (OR: 4,884; IC95% 1,199-19,890; p=0,02) e presença de complicações (OR: 5,34; IC95% 2,445 -11,770; p<0,0001). Vinte e quatro pacientes (9,45%) foram submetidos à cirurgia de colectomia total. A necessidade de cirurgia foi associada ao tempo de seguimento (OR: 1,074; IC95% 1,074-1,13; p=0,01). Seis pacientes (2,36%) apresentaram câncer colorretal. A presença de câncer colorretal foi associada com a idade ao diagnóstico (OR: 1,060; IC95%: 1,003-1,119; p=0,04) e tabagismo ativo (OR: 6,999; IC95%: 1,017-48,161; p=0,02). Vinte e cinco pacientes (9,84%) morreram. As variáveis associadas ao óbito foram a pontuação total do escore de Mayo (OR: 1,338; IC95%: 1,011-1,770; p=0,04), uso de prednisona (OR: 5,218; IC95%; 2,053-13,261; p=0,0005), presença de desnutrição (OR: 3,307, IC95%: 1,300-8.408, p=0,01) e a necessidade de hospitalização (OR: 3,307; IC95%: 1,462-28,195; p=0,01). Conclusões: a presença de pancolite e a presença de colangite esclerosante primária foram associadas à necessidade de hospitalização. A presença de câncer colorretal foi associada ao tabagismo. As variáveis associadas ao óbito foram relacionadas com a atividade da doença, como a pontuação total do escore de Mayo, o uso de prednisona, a presença de desnutrição e a necessidade de hospitalização. / Introduction: Inflammatory bowel diseases (IBD), represented by Crohn's Disease (CD) and Ulcerative Colitis (UC), can evolve with disabling symptoms that compromise the patients quality of life. The early identification of severe disease allows a more aggressive therapeutic approach with a lower risk of complications and lower rates of hospitalizations, surgeries and death. The objective of the present study was to identify the variables associated with the need for hospitalization, need for colectomy, presence of colorectal cancer and death occurrence in UC patients. Methodology: An observational and retrospective study was carried out collecting data from patients from Botucatu Medical School, totalizing 284 eligible patients. We excluded 30 patients with insufficient data, totalizing 254 analyzed. Demographic characteristics, smoking status, clinical aspects of the disease as extension and disease activity, besides presence of extraintestinal manifestations (EIM), medications in use and comorbidities were evaluated. The severity criteria considered were hospitalization due to disease complication, need for colectomy, and evolution to colorectal cancer or death. Statistical analysis: descriptive analysis and association tests. Univariate and multivariate logistic regression analyzes were performed to study the variables associated with the outcome. The outcome variables were hospitalization, colectomy, colorectal cancer and death. Survival analysis was performed using the Log Rank test, in which the initial event was the date of diagnosis and the final events were the need for hospitalization, colectomy, colorectal cancer or death or the last contact with the patient. Significance level p <0.05. The local Ethic Committee approved the study. Results: Two hundred and fifty-four UC patients were evaluated. The mean age was 46.64 (±16.88)y, 62.99% were women, 49.61% presented pancolitis and 45.68% were in clinical remission. Regarding current smoking, 66.40% of the patients were non-smokers, 28.06% ex-smokers and 5.53% smokers. EIM was observed in 52.36% of the patients and 10.63% of them was receiving biological therapy. Ninety-three patients (29.06%) required hospitalization and it was associated with pancolitis extension, presence of primary sclerosing cholangitis (OR:4.884; IC95% 1.199- 19.890; p=0.02) and presence of complications (OR:5.364; IC95% 2.445-11.770; p<0.0001). Twenty-four patients (9.45%) underwent total colectomy. The need for surgery was associated with follow-up time (OR:1.074; IC95% 1.074-1.138; p=0.01). Six patients (2.36%) presented colorectal cancer. The presence of colorectal cancer was associated with age at diagnosis (OR:1.060; 95%CI 1.003- 1.119; p=0.04) and current smoking (OR:6,999; 95%CI 1.017-48.161; p=0.02). Twenty-five patients (9.84%) died. The variables associated with death were the total Mayo Score (OR:1.338; 95%CI 1.011-1.770; p=0.04), prednisone use (OR:5.218; 95%CI 2.053-13.261; p=0.0005), presence of malnutrition (OR:3.307, 95%CI:1.300-8.408, p=0.01), and the need for hospitalization (OR:3.307; 95%CI:1.462-28.195; p=0.01). Conclusions: The presence of pancolitis and the presence of primary sclerosing cholangitis were associated with the need for hospitalization. The presence of colorectal cancer was associated with current smoking. The variables associated with death were related with disease activity, such as the total Mayo Score, prednisone use, presence of malnutrition and the need for hospitalization.

Orientador: Ligia Yukie Sassaki / Resumo: Introdução: as Doenças Inflamatórias Intestinais (IBD), representadas pela Doença de Crohn e Retocolite Ulcerativa (RCU), podem evoluir com sintomas incapacitantes que comprometem a qualidade de vida de seus portadores. A identificação precoce de doença grave permite terapêutica inicial mais agressiva com menores taxas de complicações e hospitalizações, cirurgias e morte. O objetivo do presente estudo foi identificar as variáveis associadas à necessidade de hospitalização, cirurgia de colectomia, evolução para câncer colorretal e óbito em pacientes portadores de RCU. Metodologia: foi realizado estudo observacional e retrospectivo com coleta de dados de pacientes acompanhados no Ambulatório de DII da Faculdade de Medicina de Botucatu, totalizando 284 pacientes elegíveis. Excluímos 30 pacientes com dados faltantes, totalizando 254 pacientes analisados. As características demográficas, tabagismo, aspectos clínicos como extensão e atividade da doença, além da presença de manifestações extraintestinais (MEI), medicamentos em uso e comorbidades foram avaliados. Os defechos considerados foram necessidade de hospitalização por complicações da doença, necessidade de colectomia, evolução para câncer colorretal ou óbito. Análise estatística: análise descritiva e testes de associação. Foram realizadas análises de regressão logística univariada e multivariada para avaliar as variáveis associadas ao desfecho. As variáveis de desfecho foram necessidade de hospitalização, colectomia, câncer ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Inflammatory bowel diseases (IBD), represented by Crohn's Disease (CD) and Ulcerative Colitis (UC), can evolve with disabling symptoms that compromise the patients quality of life. The early identification of severe disease allows a more aggressive therapeutic approach with a lower risk of complications and lower rates of hospitalizations, surgeries and death. The objective of the present study was to identify the variables associated with the need for hospitalization, need for colectomy, presence of colorectal cancer and death occurrence in UC patients. Methodology: An observational and retrospective study was carried out collecting data from patients from Botucatu Medical School, totalizing 284 eligible patients. We excluded 30 patients with insufficient data, totalizing 254 analyzed. Demographic characteristics, smoking status, clinical aspects of the disease as extension and disease activity, besides presence of extraintestinal manifestations (EIM), medications in use and comorbidities were evaluated. The severity criteria considered were hospitalization due to disease complication, need for colectomy, and evolution to colorectal cancer or death. Statistical analysis: descriptive analysis and association tests. Univariate and multivariate logistic regression analyzes were performed to study the variables associated with the outcome. The outcome variables were hospitalization, colectomy, colorectal cancer and death. Survival analysis was performed using the L... (Complete abstract click electronic access below) / Mestre