O comprometimento do trato gastrintestinal pela endometriose em 40 pacientes com endometriose pélvica foi avaliado pelo método da COPE. A coorte estudada compreendeu 21 pacientes com sinais e sintomas gastrintestinais e 19 pacientes sem sinais e sintomas gastrintestinais, visando a estabelecer: 1. associações e correlações entre os parâmetros clínicos que sinalizam a presença de endometriose e as localizações das lesões em cada segmento do trato gastrintestinal; 2. correlação entre o estadiamento da endometriose (ASRM, 1996) e o comprometimento gastrintestinal e 3. correlação entre a classificação histológica da endometriose e o comprometimento gastrintestinal. Através da COPE, o diagnóstico e as correlações entre sinais e sintomas ginecológicos, ultra-sonografia vaginal, classificação da ASRM e histologia, com as características distributivas da doença no trato gastrintestinal, demonstraram que: 1. A idade foi significativamente mais elevada nas pacientes com sinais e sintomas no TGI; 2. A detecção de lesões no TGI ocorreu em 70% das pacientes; 3. A dismenorréia em intensidade severa ou incapacitante e dispareunia em intensidade severa correlacionaram-se com a endometriose ginecológica e a endometriose do trato gastrintestinal na presença de sinais e sintomas no TGI; 4. Os sinais e sintomas gastrintestinais correlacionados com a endometriose ginecológica e do TGI, foram o puxo e o tenesmo cíclico, a dor em cólica cíclica, a obstipação cíclica, a diarréia cíclica, a dor pélvica acíclica, as fezes afiladas e o sangramento intestinal cíclico; 5. A endometriose que provoca sinais e sintomas no TGI está localizada no segmento retossigmóide e/ou no íleo. A dispareunia e a dismenorréia apontam para o acometimento do íleo. A infertilidade sinaliza para a endometriose no apêndice; 6. Nas correlações do toque vaginal com os achados da COPE, o aumento anexial correlaciona-se com retossigmóide e íleo, o espessamento ou nódulo ligamentar, correlaciona-se com o reto e a presença de nódulo no fundo de saco ou lesão no septo reto-vaginal, sinaliza para o acometimento do cólon sigmóide; 7. A COPE aplicada a pacientes com suspeita de endometriose no segmento retossigmóide levantada pela ultra-sonografia transvaginal mostra que a doença se estende em associação significante ao reto, ao cólon sigmóide, cólon ascendente e íleo; 8. A COPE demonstrou que a endometriose acomete mais freqüentemente o íleo, o apêndice, o segmento retossigmoideano e o cólon ascendente na existência ou não de sinais e sintomas do TGI; 9. Os padrões histológicos distribuíram-se igualmente pelas lesões endometrióticas do TGI exceto nas lesões do mesentério; 10. O estádio IV da ASRM, 1996, é fator de risco significativo para o acometimento do reto, sigmóide e íleo. / The importance of the laparoscopic procedure padronization to the run in the peritoneal organs was established in 40 patients with pelvic endometriosis through the observation of its efficient in the characterization of the gastrointestinal tract endometriosis distribution and in the establishment of the meaningful correlations among the most important aspects of the gynecological clinic, vaginal ultrasonagraphic exam, ASRM and histology with gastrointestinal signs and symptoms. The run in the peritoneal organs in the endometriosis has permitted the diagnosis of the endometriosis disease real extension if compared to the competing methods previously described in the pertinent literature

A distrofia muscular de Duchenne (DMD) é uma doença neuromuscular fatal, a mais comum das distrofias musculares, causada pela ausência da proteína distrofina, componente importante do complexo glicoproteínadistrofina, que envolve as células musculares lisas, cardíacas e esqueléticas, estabilizando a membrana dessas células durante contrações e relaxamentos. Por ser uma doença genética ligada ao cromossomo X, acomete indivíduos do sexo masculino, sendo que casos raros de meninas afetadas também são relatados. Indivíduos afetados pela DMD apresentam alterações significantes na composição corpórea, comparados com a população normal. Cães da raça golden retriever que possuem distrofia muscular (GRMD) representam o melhor modelo animal para ensaios terapêuticos na busca do tratamento da DMD, sendo esses animais fundamentais para o estudo do desenvolvimento dessa anomalia, devido ao fato de apresentarem genes e sintomas clínicos homólogos aos dos pacientes humanos. A similaridade entre as duas doenças é surpreendente, ambas são caracterizadas por precoce miopatia degenerativa, com progressiva necrose, fagocitose, deposição de cálcio e fibrose endomisial e perimisial. O foco principal deste trabalho foi esclarecer dúvidas quanto ao comportamento da musculatura lisa dos animais distróficos e portadores da GRMD, para se conhecer os efeitos da doença nesse tipo de tecido. Para avaliação da musculatura lisa uterina foram utilizados quatro úteros de cadelas golden retriever saudáveis, cinco úteros de portadoras do gene e quatro úteros de animais afetados pela GRMD. Analisando as fotomicrografias uterinas dos grupos, podemos notar que não há diferenças morfológicas através dos estudos histológicos convencionais. Porém, quando analisamos as fotomicrografias polarizadas , nota-se nas cadelas saudáveis exclusivamente colágeno do tipo III, de coloração esverdeada. Quanto às portadoras do gene da GRMD, observa-se colágeno predominantemente vermelho, do tipo I, e pouco em verde, tipo III. Fotomicrografias das fêmeas afetadas pela GRMD encontraram colágeno tipo III na parte interna do endométrio, miométrio e perimétrio, e colágeno tipo I em epitélio vascular, porém ambos em menor quantidade. É possível visualizar colágeno entre as fibras musculares através da luz polarizada. Para a avaliação da digestibilidade dos nutrientes em cães GRMD e sua capacidade absortiva, foram utilizados nove cães GRMD, sete fêmeas portadoras e dez animais saudáveis; todos receberam a mesma dieta adicionada de oxido crômico. O período experimental constou de um intervalo de 10 dias, sendo os cinco primeiros dias de adaptação e os cinco restantes de coleta de amostras. Os coeficientes de digestibilidade aparente (CDAs) obtidos pelos grupos estudados foram comparados e analisados: os resultados indicaram que os cães com GRMD avaliados não apresentaram redução do aproveitamento nutricional. Novas pesquisas que avaliem o gasto energético e composição corpórea destes animais são necessárias para melhor compreensão dos mecanismos envolvidos no emagrecimento e perda de peso que apresentam. Para a avaliação do tempo de trânsito gastrintestinal (TTGI) em cães golden retriever saudáveis, portadores e afetados pela distrofia muscular, foram utilizados 18 cães divididos em três grupos animais saudáveis, portadores e afetados pela GRMD, com seis cães cada. Os animais foram acondicionados em canis individuais e receberam a mesma dieta; esferas de polietileno (BIPS) impregnadas com bário foram fornecidas durante a alimentação. Cada animal recebeu 10 marcadores de tamanho grande (5 mm de diâmetro) e 30 marcadores de tamanho pequeno (1.5mm de diâmetro). Os animais foram radiografados nos momentos antes (basal) da 1ª refeição, para confirmação da ausência de conteúdo alimentar e fecal no trato gastrintestinal, e após a alimentação, em intervalos de duas horas, até que os marcadores atingissem o cólon. Resultados apontaram que os cães GRMD avaliados apresentam TTGI mais rápido quando comparados aos demais grupos. Este trabalho comprovou que a mm. lisa de cães distróficos é, de diversas maneiras, afetada pela doença, apresentando tanto alterações morfológicas como morfofuncionais. Muitos estudos nessa área são necessários para melhor compreensão dos mecanismos da enfermidade, bem como maneiras de intervenção para melhoria da qualidade de vida dos indivíduos afetados. Cães GRMD(s) são o melhor modelo animal para estudos pré-clínicos da DMD, assim esses resultados são muito relevantes para aprimoramento e melhora de manejo em biotérios que abrigam esses animais e para extrapolação de dados na própria medicina / Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disease, it is the most common muscular dystrophy caused by absence of dystrophin protein, an important component of the dystrophin-glycoprotein complex, involving cell membranes of smooth, cardiac and skeletal cells, stabilizing these cells during contractions and relaxations. Because it is a genetic disorder linked to the X chromosome, it affects males, with rare cases of girls affected also reported. Individuals affected by DMD show significant changes in body composition, compared with the normal population. Golden retriever dogs who have muscular dystrophy (GRMD) represent the best animal model for therapeutic trials in the treatment of DMD,; these animals are fundamental to the study of the development of this anomaly, due to the fact that they show clinical symptoms and genes homologous to the human patients. The similarity between the two diseases is staggering; both are characterized by early degenerative myopathy with progressive necrosis, phagocytosis, calcium deposition and endomysial and perimysial fibrosis. The main focus of this thesis was to answer questions about the behavior of smooth muscles of dystrophic animals and carriers of GRMD, to know the effects of the disease in this tissue type. For evaluation of uterine smooth muscle we utilized four uteri of golden retriever healthy bitches, five uteri of gene carriers and four uteri of animals affected by GRMD. Analyzing the uterine photomicrographs of the groups we note that there are no morphological differences in conventional histological studies. However, when we analyze the polarized photomicrographs, we note that in healthy bitches we can see only collagen type III of greenish color. As regards the carriers of the GRMD gene, there is predominantly red collagen , type I, and very little green, type III. Photomicrographs of females affected by GRMD found type III collagen inside the endometrium, myometrium and perimetrium and collagen type I in vascular epithelium, but both in a very little quantity. It is possible to visualize collagen between muscle fibers by polarized light. For the assessment of nutrient digestibility in dogs GRMD and its absorptive capacity, we used nine GRMD, seven carriers females and ten healthy animals and all of them received the same diet with added chromic oxide. The experimental period consisted of an interval of 10 days, with the first five days to adaptation and the last ones to sample collection. The apparent digestibility coefficients obtained by the groups were compared and analyzed; the results showed that dogs with GRMD evaluated showed no reduction in energy utilization. The experimental period consisted of an interval of 10 days, with the first five days to adaptation and the last of them to sample collection. The apparent digestibility coefficients obtained by the groups were compared and analyzed; the results showed that dogs with GRMD evaluated showed no reduction in nutritional absorption. New research to assess energy expenditure and body composition in these animals are needed to a better understanding of the mechanisms involving reduction and weight loss they present. For the assessment of gastrointestinal transit time (TTGI) in golden retriever dogs healthy carriers and affected by GRMD 18 dogs divided into three groups were used - healthy animals, carriers and affected by GRMD, with six dogs each. The animals were placed in individual cages and were fed the same diet; polyethylene spheres (BIPS) impregnated with barium were supplied during feeding. Each animal received 10 markers large size (5mm diameter) and 30 markers small size (1.5mm diameter). Radiographs were taken in the moments before (baseline) the 1 st meal, to confirm the absence of food and fecal content in the gastrointestinal tract and after feeding at two-hour intervals until markers reach the colon. Results indicated that GRMD dogs evaluated showed TTGI faster when compared to the other groups. This thesis has shown that the smooth muscles of dystrophic dogs are, in many ways, affected by the disease, with both morphological and morphofunctional alterations; many studies in this area are needed so we can better understand the mechanisms of disease, as well as ways of intervention to improve the quality of life of affected individuals. GRMD dogs are the best animal model for preclinical studies of DMD, so these results are very relevant to enhancement and improvement of management in bioteries these animals have

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-11-24T17:44:53Z No. of bitstreams: 2 Dissertação - Luciana Marya Gusmão Tartuce - 2016.pdf: 2825829 bytes, checksum: fe4eb785221991a505a52e87e318ad9a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2016-11-28T17:52:09Z (GMT) No. of bitstreams: 2 Dissertação - Luciana Marya Gusmão Tartuce - 2016.pdf: 2825829 bytes, checksum: fe4eb785221991a505a52e87e318ad9a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-11-28T17:52:09Z (GMT). No. of bitstreams: 2 Dissertação - Luciana Marya Gusmão Tartuce - 2016.pdf: 2825829 bytes, checksum: fe4eb785221991a505a52e87e318ad9a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-10-27 / The changing profile of morbidity and mortality points to an increase in both life expectancy of human beings and the growing prevalence of chronic degenerative diseases, such as cancer. Regarding the impact that both diagnosis and treatment, especially, surgical one, have on patients, their families, and social environment, it is worth caring about the overall health (OH) and Quality of Life (QOL) as perceived by the individual. The main objective of this study was to assess health-related QOL among patients undergoing surgeries for gastrointestinal cancer who were treated in a surgery clinic of a school hospital. It is a descriptive exploratory quantitative study, carried out with 31 patients, mostly women, from the capital of Goiás, aged between 60 and 69 years old, catholic, married, retired, with elementary level of education, predominantly diagnosed with colorectal cancer, all of them treated at the Surgery Clinic of the HC/UFG. The instruments used were: Socio-demographic Questionnaire, QSG, WHOQOL-bref and EORTC QLQ-C30. The socio-demographic data and diagnosis were assessed regarding its frequency and average. The WHOQOL-bref, EORTC QLQ-C30 and QSG were assessed according to their manuals and guidelines. The data collected was organized in the SPSS 18.0 statistical program. The quality of life was considered to be regular in two moments (30 and 60 postoperative days), being the Physical domain the most affected. As for the overall health, the QSG showed some effect in the sleep disturbance factor in the 30-day assessment, and in the 60-day assessment all factors were considered normal. The correlation between the factors of OH and the EORTC QOL scores C-30 showed positive and significant impact in all dimensions: the better overall health, the better QOL. However, in the correlation of the OH and the QOL assessed through the WHOQOL-Bref, there was only significant and positive correlation between OH and the Physical Dimensions and the environment. It is concluded that assessing the QOL of sick people is a complex process due to the characteristics of subjectivity and multidimensionality involved, making it necessary the use of valid and reliable instruments. The surgical procedure has brought benefits to the participants of this study, however qualitative methodologies could be added to the understanding of the meanings as assigned by the sick people, with similar experiences of cancer treatment, to several factors of their QOL and their OH, throughout the follow-up. Thus, the evaluation of the perception of QOL and the OH can be inserted in clinical practice and regular care of sick people. / A mudança do perfil de morbi-mortalidade aponta aumento da expectativa de vida do ser humano e crescimento da prevalência das doenças crônico-degenerativas, tais como o câncer. Considerando o impacto que este diagnóstico e seu tratamento, principalmente o cirúrgico, exercem sobre o enfermo e seu meio social e familiar, há de se preocupar com a Saúde Geral (SG) e a Qualidade de Vida (QV) percebida pelo indivíduo. O objetivo geral do estudo foi avaliar a qualidade de vida relacionada à saúde de pacientes operados por câncer do aparelho gastrointestinal assistidos na clínica cirúrgica de um hospital universitário. Trata-se de um estudo descritivo, exploratório e quantitativo, realizado com 31 pacientes, em sua maioria mulheres, provenientes da capital de Goiás, com faixa etária entre 60 e 69 anos, casados, aposentados, católicos, nível de escolaridade fundamental, com diagnóstico predominante de câncer colorretal, atendidos na clínica cirúrgica do HC/UFG. Os instrumentos utilizados foram: Questionário Sociodemográfico, QSG, WHOQOL-bref e EORTC QLQ-C30. Os dados sociodemográficos e o diagnóstico foram computados em sua frequência e média. O WHOQOL-bref, o EORTC QLQ-C30 e QSG foram avaliados de acordo com seus manuais de recomendação. Os dados coletados foram sistematizados no programa estatístico SPSS, versão 18.0. A qualidade de vida foi considerada regular nos dois momentos avaliados (30 e 60 dias pós-cirúrgico), sendo o domínio Físico o mais afetado. Quanto à saúde geral, o QSG apontou comprometimento no fator distúrbio do sono na avaliação de 30 dias, sendo que na avaliação de 60 dias todos os fatores já estavam dentro da normalidade. A correlação entre os fatores de SG e escores de QV da EORTC C-30 mostrou haver impacto positivo e significativo em todas as dimensões: quanto melhor avaliada a saúde geral, melhor foi avaliada a QV. Contudo, na correlação da SG e a QV avaliada por meio do WHOQOL-Bref, apenas houve correlação significativa e positiva entre SG e as Dimensões Física e Meio Ambiente. Concluiu-se que avaliar a QV de enfermos é um processo complexo pelas características de subjetividade e multidimensionalidade envolvidas, fazendo-se necessário o uso de instrumentos válidos e confiáveis. O procedimento cirúrgico trouxe benefícios aos participantes deste estudo sendo que metodologias qualitativas poderiam ser acrescidas para a compreensão dos sentidos atribuídos pelos enfermos, com vivências similares de tratamento do câncer, aos diversos fatores de sua QV e de sua SG, ao longo do follow-up. A avaliação da percepção da QV e da SG pode ser inserida na prática clínica e nos cuidados regulares dos enfermos.

Submitted by Edineia Teixeira (edineia.teixeira@unioeste.br) on 2017-12-18T17:12:18Z No. of bitstreams: 2 Angélica_capelassi2016.pdf: 1330041 bytes, checksum: 1450f27720ae9da87b224a8f974b2a87 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-12-18T17:12:18Z (GMT). No. of bitstreams: 2 Angélica_capelassi2016.pdf: 1330041 bytes, checksum: 1450f27720ae9da87b224a8f974b2a87 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-06-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Obesity is a risk factor in the development of many diseases such as diabetes and hypertension. In addition, it is associated with the development of diseases in the gastrointestinal tract (GIT) such as gastroesophageal reflux and gastritis. Bariatric surgery has proved to be the only effective long-term treatment modality sustained effects for the treatment of morbid obesity and its comorbidities. Studies have shown that bariatric procedures affect the morphology and function of the GIT. Few studies have investigated the effects of sleeve gastrectomy (SG), a restrictive technique that reduces the size of the stomach on the GIT. Thus, our objective was to analyze the effect of SG on gastric histopathology and the morphology of the small intestine (duodenum, jejunum and ileum) in obese rats fed a cafeteria diet. For this study, 8-week-old Wistar rats were divided into two groups: control (CTL), which received standard diet, and cafeteria (CAF), which received cafeteria diet to induce obesity. After two months, the CAF group underwent pseudo-surgery (CAF PS) or sleeve gastrectomy (CAF SG). At three months after surgery obesity was verified and stomach, duodenum, jejunum and ileum were collected and analyzed. Body weight and retroperitoneal and perigonadal fat pads were higher in CAF PS animals compared to the CTL. The SG did not influence these parameters. Regarding the morphology of the stomach, we observed that the CTL animals showed normal morphology of this organ, while the CAF PS animals showed changes in the gastric mucosa with the presence of hyperemia, mild inflammatory infiltrate and diffuse in the mucosa and submucosa, as well as mild erosion of the gastric mucosa and atrophy. The SG exacerbated changes in the stomach. CAF SG animals showed severe erosion of the gastric mucosa, edema, moderate and diffuse inflammatory infiltrate in the mucosa and submucosa, as well as atrophy of the muscular layer and the body of the mucosa. In relation to morphometry of the small intestine, no changes were found in the duodenum, jejunum and ileum of CAF PS and CTL animals. However, CAF SG animals showed increase in total thickness of the duodenum, as well as increase in the thickness of the mucosa and submucosa layer and villi. The jejunum and ileum showed no change. In the summary cafeteria diet causes changes in the gastric mucosa without changing the small intestine. The SG, three months after the procedure, exacerbates gastric alterations and promotes changes in the duodenum morphometry. / A obesidade é um fator de risco para o desenvolvimento de várias doenças, tais como diabetes e hipertensão. Além disso, está associada com o desenvolvimento de doenças no trato gastrointestinal (TGI), como refluxo gastroesofágico e gastrite. A cirurgia bariátrica tem provado ser a única modalidade de tratamento eficaz com efeitos sustentados por longo prazo para o tratamento da obesidade mórbida e suas comorbidades. Estudos demonstram que os procedimentos bariátricos alteram a morfologia e o funcionamento do TGI. Poucos estudos investigam os efeitos da gastrectomia vertical (GV), uma técnica restritiva que reduz o tamanho do estômago, sobre o TGI. Dessa forma, nosso objetivo foi analisar o efeito da GV sobre a histopatologia gástrica e a morfologia do intestino delgado (duodeno, jejuno e íleo) em ratos obesos pela dieta de cafeteria. Para este trabalho, ratos Wistar com 8 semanas de vida foram divididos em dois grupos: controle (CTL), que recebeu dieta padrão, e cafeteria (CAF), que recebeu dieta de cafeteria para indução da obesidade. Após dois meses, o grupo CAF foi submetido à pseudo-cirurgia (CAF PC) ou à gastrectomia vertical (CAF GV). Três meses após o procedimento cirúrgico foi realizada a avaliação da obesidade e o estômago, duodeno, jejuno e íleo foram coletados e analisados. O peso corporal e o peso das gorduras retroperitonial e perigonadal foi maior nos animais CAF PC em comparação com os CTL. A GV não influenciou estes parâmetros. Em relação a morfologia do estômago, observamos que os animais CTL apresentaram morfologia normal desse órgão, enquanto os animais CAF PC apresentaram alterações na mucosa gástrica com presença de hiperemia, infiltrado inflamatório leve e difuso na mucosa e submucosa, bem como.leve erosão da mucosa gástrica e atrofia. A GV exacerbou as alterações no estômago. Os animais CAF GV apresentaram erosão intensa da mucosa gástrica, edema, infiltrado inflamatório moderado e difuso na mucosa e submucosa, bem como atrofia da camada muscular e da mucosa do órgão. Em relação a morfometria do intestino delgado, nenhuma alteração foi encontrada no duodeno, jejuno e íleo entre os animais CAF PC e CTL. Entretanto, os animais CAF GV apresentaram aumento da espessura total do duodeno, bem como, aumento na espessura da camada mucosa e submucosa e na altura das vilosidades. O jejuno e o íleo não apresentaram modificações. Assim, concluímos que, a dieta de cafeteria promove alterações na mucosa gástrica sem modificar o intestino delgado. A GV, três meses após o procedimento, exacerba as alterações gástricas e promove modificações na morfometria do duodeno

Made available in DSpace on 2017-07-10T17:48:17Z (GMT). No. of bitstreams: 1 Josiane_Mangoni.pdf: 304825 bytes, checksum: 57fa6d5d799ce082b0c419c954f47047 (MD5) Previous issue date: 2009-06-29 / Fundação Araucária / The use of probiotic in animal nutrition has been identified for reducing mortality resulting from intestinal colonization by pathogenic micro-organisms, improving the performance and characteristics of production, leaving no residue in meat. The experiment aimed to isolate, from faeces samples from swine in the suckling strains of lactobacilli in order to be identified as probiotics. 92 colonies were isolated, these pre-selected colonies were subjected to the catalase which 60 negatively. The 60 colonies were subjected to negative Gram stain were evaluated where their morphology. Of these, 16 had forms of bacilli. These isolates were compared for their ability to resist pH 3.0, to grow in the presence of bile salts, phenol, lysozyme, its ability to hydrophobicity and antagonistic. Isolates L03, L04, L08 and L15, identified in this study show better for probiotic use, according to show better performance on the acidic conditions, growing in the presence of bile salts and phenol, with high percentage of hydrophobicity and inhibiting Escherichia coli / O uso de probiótico na alimentação animal tem sido indicado, por reduzir a mortalidade resultante da colonização intestinal por micro-organismos patógenos, melhorar o desempenho e as características de produção sem deixar resíduos na carne. O experimento teve como objetivo isolar, a partir de amostras de fezes de suínos na fase de aleitamento, cepas de lactobacilos visando sua utilização como probióticos. Foram isoladas 92 colônias, essas colônias pré-selecionadas foram submetidas à prova de catalase onde 60 negativamente. As 60 colônias negativas foram submetidas à coloração de Gram onde foram avaliadas sua morfologia. Destas, 16 apresentaram formas de bacilos. Esses isolados foram comparados pela sua habilidade em resistirem em pH 3,0, crescer na presença de sais biliares, fenol, lisozima, sua capacidade de hidrofobicidade e antagônica. Os isolados L03, L04, L08 e L15, identificados neste trabalho apresentam melhores características para uso como probiótico, em função de demonstrar melhor comportamento sobre as condições ácidas, crescendo na presença de sais biliares e fenol, apresentando alta percentagem de hidrofobicidade e inibindo Escherichia coli

Antecedentes y propósito del estudio: la hemorragia digestiva alta es una causa importante de ingreso hospitalario y constituye la principal emergencia gastroenterológica, con una tasa de mortalidad de hasta el 14%. En el Perú no existen estudios sobre el uso de la escala de Glasgow-Blatchford para predecir mortalidad por hemorragia digestiva alta. El objetivo de este estudio es realizar la validación externa de la escala de Glasgow-Blatchford y establecer su mejor punto de corte para predecir mortalidad por hemorragia digestiva alta en un hospital de Lima, Perú. Métodos: estudio de validación diagnóstica, analítico, longitudinal, de tipo retrospectivo, con datos de pacientes con diagnóstico clínico y endoscópico de hemorragia digestiva alta atendidos en la Unidad de Hemorragia Digestiva del Hospital Nacional Edgardo Rebagliati Martins, entre junio de 2012 y diciembre de 2013. Calculamos el área bajo la curva ROC (receiver operating characteristic) de la escala de Glasgow-Blatchford para predecir mortalidad, con un intervalo de confianza al 95%. Resultados: un total de 339 registros fueron analizados. El 57,5% fueron varones y la edad media (desviación estándar) fue de 67,0 (15,7) años. La mediana de la escala de Glasgow-Blatchford obtenida en la población fue de 12. El análisis ROC para mortalidad dio un área bajo la curva de 0,59 (IC95% 0,5-0,7). Se estratificó por tipo de hemorragia digestiva alta, obteniendo un área bajo la curva de 0,66 (IC95% 0,53-0,78) para el tipo no variceal. Conclusiones: en la población estudiada, la escala de Glasgow-Blatchford no posee una validez diagnóstica adecuada para predecir mortalidad.

Introducción: El sangrado digestivo bajo (SDB) es una entidad cuyas tasas de complicaciones y mortalidad se han incrementado en las últimas décadas. Si bien se han identificado algunos factores relacionados a mal pronóstico, aún quedan variables por evaluar. Objetivo: Identificar factores de mal pronóstico en pacientes que presentaron SDB en el Hospital Nacional Edgardo Rebagliati Martins de Lima, Perú. Materiales y métodos: Se realizó un estudio observacional analítico de tipo cohorte retrospectivo. Se realizó un censo de todos los pacientes que presentaron SDB agudo entre Enero 2010 y Diciembre 2013. Las variables principales a evaluar fueron frecuencia cardiaca ≥ 100/min, presión arterial sistólica < 100 mmHg y hematocrito bajo (≤ 35%) al ingreso. Se definió mal pronóstico como cualquiera de los siguientes criterios: muerte durante la hospitalización, sangrado que requiera transfusión de ≥ 4 unidades de sangre, reingreso dentro del primer mes, o necesidad de cirugía de hemostasia. Resultados: Se incluyó un total de 341 pacientes con SDB, de los cuales el 27% tuvo mal pronóstico y 2% fallecieron. Se encontró como variables asociadas a mal pronóstico: frecuencia cardiaca ≥ 100/min al ingreso (RR: 1,75 IC 95% 1,23-2,50), presión arterial sistólica < 100 mmHg al ingreso (RR: 2,18 IC 95% 1,49-3,19), hematocrito ≤ 35% al ingreso (RR: 1,98 IC 95% 1,23-3,18) y sangrado de origen no determinado (RR: 2,74 IC 95% 1,73-4,36). Conclusiones: Frecuencia cardiaca elevada al ingreso, hipotensión sistólica al ingreso, hematocrito bajo al ingreso y presentar un sangrado en el cual no se encuentra el punto de origen son factores que incrementan el riesgo de presentar mal pronóstico, por lo que se recomienda un monitoreo más estricto en estos pacientes. / Background: Lower gastrointestinal bleeding (LGIB) is an event that has shown an increase in complications and mortality rates in the last decades. Although some factors associated with poor outcome have been identified, there are several yet to be evaluated. Objective: To identify risk factors for poor outcome in patients with LGIB in the Hospital Nacional Edgardo Rebagliati Martins of Lima, Peru. Material and methods: A prospective analytic observational cohort study was made, and a census was conducted with all patients with acute LGIB between January 2010 and December 2013. The main variables were heart rate ≥ 100/min, systolic blood pressure < 100 mmHg and low hematocrit (≤ 35%) at admission. Poor outcome was defined as any of the following: death during hospital stay, bleeding requiring transfusion of ≥ 4 blood packs, readmission within one month of hospital discharge, or the need for hemostatic surgery. Results: A total of 341 patients with LGIB were included, of which 27% developed poor outcome and 2% died. Variables found to be statistically related to poor outcome were: heart rate ≥ 100/min at admission (RR: 1,75 IC 95% 1,23-2,50), systolic blood pressure < 100 mmHg at admission (RR: 2,18 IC 95% 1,49-3,19), hematocrit ≤ 35% at admission (RR: 1,98 IC 95% 1,23-3,18) and LGIB of unknown origin (RR: 2,74 IC 95% 1,73-4,36). Conclusions: Elevated heart rate at admission, systolic hypotension at admission, low hematocrit at admission and having a LGIB of unknown origin are factors that increase the risk of developing poor outcome, and these patients should be monitored closely due to their higher risk of complications.

The human GI tract has evolved to simultaneously absorb nutrients and be the frontline in host defence. These seemingly mutually exclusive goals are achieved by a single cell thick epithelial barrier, and a complex resident immune system which lives in symbiosis with the intestinal microflora and is also able to rapidly respond to invading pathogens. An immunological balance is therefore required to permit tolerance to the normal intestinal microflora, but also prevent the dissemination of pathogenic micro-organisms to the rest of the host. Inappropriate immune responses in genetically susceptible individuals are the hallmark of human inflammatory bowel disease (IBD) and are thus targeting effector immune cells and their cytokines remains the mainstay of treatment. However despite vigorous efforts to delineate the genetic contribution to IBD disease susceptibility using large multinational cohorts, the majority of disease heritability remains unknown. Epigenetics describes heritable changes in chromatin that are not conferred by DNA sequence. These incorporate changes to histones, chromatin structure and DNA methylation, which confer changes to gene transcription and thus gene expression and cellular function. Methylbinding proteins (MBD) have the ability to bind to methylated DNA and recruit large chromatin remodeling complexes that underpin a variety of epigenetic modifications. Methyl- CpG-binding domain protein 2 (MBD2) is one such MBD that is required for appropriate innate (dendritic cell) and adaptive (T cell) immune function, though its role has not been investigated in the GI tract. We hypothesized that alterations in chromatin are central to the reprogramming of normal gene expression that occurs in disease states. By defining the phenotype of immune cells in the absence of MBDs we hope to understand the mechanisms of chromatin-dysregulation that lead to immune-mediated diseases such as IBD. We therefore aimed to assess the role of MBD2 in colon immune cells in the steady state and in murine models of GI tract inflammation, thereafter identifying the culprit cell types and genes responsible for any observed changes. We envisaged that investigating heritable, epigenetic changes in gene expression that are inherently more amenable to environmental manipulation than our DNA code, may provide novel insight to a poorly understood mechanism of disease predisposition. In addition identifying the cellular and gene targets of Mbd2 mediated changes to immune homeostasis that may provide exciting and novel approaches to therapeutic modulation of pathological inflammatory responses. In chapter 3 we assessed the expression of Mbd2/MBD2 in the murine/human GI tract. Consistent with existing mouse data, levels of Mbd2 mRNA increased between anatomical divisions of small (duodenum, ileum, terminal ileum) and large intestine (caecum, colon, rectum). In addition MBD2 mRNA was greater in the rectum versus ileum, with active IBD associated with lower rectal MBD2 mRNA compared to quiescent IBD controls. Thus we sought to understand the role of Mbd2 in the colon, where mRNA levels were the highest in the GI tract and where appropriate immune function is central to prevent damaging inflammation. To address these aims required the development of existing methods of cell surface marker expression analysis using flow cytometry techniques to simultaneously identify multiple innate and adaptive immune populations. Using naïve Mbd2 deficient mice (Mbd2-/-) we observed CD11b+ CD103+ DCs were significantly reduced in number in Mbd2 deficiency. To understand the role of Mbd2 in colonic inflammation we employed a mouse model of chemical (DSS) and infectious (T. gondii) colitis comparing Mbd2-/- and littermate controls (WT). Mbd2-/- were extremely sensitive to DSS and T. gondii mediated colonic inflammation, characterized by increased symptom score, weight loss and histological score of tissue inflammation (DSS) and increased antibody specific cytokine responses (T. gondii) in Mbd2 deficient animals. Flow cytometry analysis of colon LP cells in both infectious and chemical colitis revealed significant accumulation of monocytes and neutrophils in Mbd2-/-. Indeed monocytes and neutrophils were the principal myeloid sources of IL-1b and TNF in DSS colitis and the number of IL-1b/TNF+ monocytes/neutrophils was significantly greater in Mbd2-/-. Lastly we employed our colon LP isolation techniques to analyse immune populations in active and quiescent IBD and healthy controls, using endoscopically acquired biopsy samples. Analysis revealed that as in murine colitis, active human IBD is characterized by the accumulation of CD14High monocyte-like cells, with an associated increased ratio of macrophage:monocyte-like cells. In Chapter 4 we sought to understand the cellular sources of Mbd2 that may explain the predisposition of Mbd2-/- to colitis. Firstly we restricted Mbd2 deficiency to haematopoietic cells using grafting Mbd2-/- bone marrow (BM) into lethally irradiated WT mice. These animals treated with DSS displayed increased weight loss, symptom score, neutrophil accumulation and histopathology score compared to mice irradiated and grafted with WT BM. Given the accumulation of monocytes in Mbd2-/- DSS treated mice, and existing literature supporting a pathogenic role in this model, we then investigated the role of Mbd2 in monocyte function. Colon monocytes sorted from Mbd2-/- and WT DSS treated mice displayed similar expression for many pro-inflammatory genes (Il6, Il1a, Il1b, Tnf), but demonstrated significantly dysregulated expression for some others (Regb, Lyz1, Ido1, C4a). To investigate this in a more refined model, we lethally irradiated WT mice and repopulated them with a WT:Mbd2-/- BM mix. This enabled the analysis of WT and Mbd2-/- haematopoietic cells in the same animal. Colon WT and Mbd2-/- monocyte recruitment and cytokine production in DSS treated mixed BM chimeras was equivalent between genotypes suggesting that Mbd2 deficiency in monocytes alone did not explain the increased susceptibility of Mbd2-/- to DSS colitis. We then restricted Mbd2 deficiency to CD11c expressing cells, given the known role for Mbd2 in their function, and for CD11c+ cells in DSS, using a CD11cCreMbd2Fl/Fl system. DSS treated mice with Mbd2 deficient CD11c+ cells demonstrated increased weight loss, symptoms score, histolopathology score, monocyte and neutrophil colon accumulation compared to controls. To further explore the role of Mbd2 in colon CD11c+ cells, macrophage and DCs from DSS treated WT and Mbd2-/- mice were purified and their gene expression analysed. Mbd2-/- versus WT macrophages demonstrated significantly altered expression of both pro- (Il1a, C6, Ido1, Trem2) and antiinflammatory (Tgfbi, Retnla) pathways that we hypothesized was a method for attempted host control of excessive colon damage in Mbd2-/- mice. DC gene expression analysis was hampered by small sample size, but demonstrated a large number of small expression changes, including IL-12/IL-23 (Jak2) and autophagy (Lrrk2) pathways. Lastly levels of costimualtory molecules (CD40/CD80) were increased in Mbd2-/- but not CD11cΔMbd2 colon LP DCs/macrophages suggesting that non-CD11c+ cellular sources of Mbd2 were required to produce increased activation phenotype in these cells. Finally in Chapter 5 we explored the role for Mbd2 in non-haematopoietic cells, namely the colonic epithelium. Here we first developed a novel method for identifying and purifying these cells using flow cytometry. Mbd2 deficient colonic epithelium demonstrated increased expression of activation markers MHC II and LY6A/E in the steady state and in DSS / T. muris mediated colonic inflammation. Indeed FACS purified colon epithelial cells from naive and DSS treated, Mbd2-/- and WT mice revealed conserved dysregulated gene expression independent of inflammation: Both naïve and inflamed Mbd2 deficient epithelium displayed significantly increased expression of genes responsible for antigen processing/presentation (MHC I, MHC II, immunoproteasome) and decreased expression of genes involved in cell-cell adhesion (Cldn1, Cldn4). Lastly we investigated whether the observed differences in Mbd2-/- cell types conferred alterations in the makeup of the intestinal microflora. Interestingly independent of co-housing of Mbd2-/- and WT animals, Mbd2 deficiency consistently predicted the microbial composition, with increased levels of Clostridales and decreased levels of Parabacteroides bacteria. Collectively we have identified CD11c+ cells, monocytes and colon epithelial cells as key cell types for Mbd2 mediated changes in gene expression that affect mucosal immune responses. These data thus identify Mbd2 gene targets within these cell types as exciting new areas for investigation and therapeutic modulation to limit damaging GI tract inflammation.

The scope of this masters thesis is the preparation of a model mucin system and its utilization as an artificial model of the native mucus system. The creation of this model system, according to several designed methods was a part of experimental part of the thesis. The preparation of mucin system comprised of physical and chemical methods of hydrogel formation, screening and characterisation of the various physical conditions of the mucin properties on its molecular level, and the preparation of sorbent with sorption surface containing mucin. Methods of light scattering, namely dynamic light scattering (DLS), used for mucin particles size change monitoring, and electroforetic light scattering (ELS), used for Zeta potential change monitoring, were used for the screening of the impact of physical factors on the properties of mucin.For the characterisation of impact of the temperature on changes in mucin sctructure was, apart from monitoring of light scattering, used also a diferential scanning calorimetry (DSC), which registered temperature value, at which mucin thermal denaturation occurs. In the next part of the thesis we subdued the created sorption surfaces to various physical-chemical analyses, which task is the characterisation and projection of surface and confirmation of mucin presence.Substancial part in monitoring and characterisation of changes in surface sctructure of sorption surface was accomplished by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscophy (SEM) was used for the final, more detailed, projection of the mucin enriched, sorbent surface structure. Suggested methods of mucin hydrogel, didnt prove sufficient results for the possibility of application of hydrogel as a artificial model of real mucus system, but the sorbent application was indicated as a suitable alternative and an instrument for the further mucin behaviour research and possibly subsequent bacterial adhesion, which represents the first step in the formation of the bacterial biofilm.

Human milk contains ~ 25 µg/mL of soluble cluster of differentiation 14 (sCD14) protein, a pattern recognition receptor (PRR) that triggers the innate immune system to respond to bacterial lipopolysaccharide (LPS). To date, the role of CD14 in the digestive tract of breast fed infants has not been well characterized and is the subject of this thesis. To investigate the biodistribution of proteins such as CD14 in vivo, a novel method for 14C radiolabeling of proteins to high specific radioactivity was developed using in vacuo methylation. Bovine serum albumin (BSA) and casein were used as test proteins to determine the following: 1) The efficacy of the in vacuo radiolabeling procedure; 2) The extent of incorporation of the 14C-label into the organs of oro-gastric gavaged 10 day old Sprague Dawley rats. [14C]BSA, [14C]casein and [14C]CD14 were prepared with specific radioactivities of 10 400, 10 800 and 163 000 dpm/µg, respectively. After feeding 6.25 µg of 14C-labeled proteins, quantifiable levels of 14C were found in the stomach, jejunum, duodenum, ileum, large intestine, intestinal luminal flushes, blood, liver, spleen and kidneys of rats. The accumulation of radiolabel in the organs of [14C]CD14 fed rats was temporally and spatially distinct from [14C]BSA and [14C]casein. Most notably, the label persisted in the stomach 480 min post-gavage. To design a neonate animal model for biodistribution, the segmental and total gastrointestinal transit times (GItt) were measured in two litters of 10 and 15 day old Sprague Dawley rat pups using barium sulfate. Ten day old rat pups that remained with and without the dam had a total gastrointestinal transit time of 13.8 ± 0.9 hr and 9.3 ± 0.7 hr, respectively. This decrease (p<0.05) in total gastrointestinal transit time in the absence of the dam was age dependent, as it was not observed (p>0.05) in the 15 day old rat pup litter. The immunological impact of an exogenous sCD14 source was examined in human peripheral blood mononuclear cells (PBMC). Pre-treatment of CD14+ monocytes with sCD14 had a protective effect, one of reducing the production of proinflammatory cytokines (TNF-α, IL-6, IL-8, IL-1β) when challenged with LPS. 14C was absorbed by neonate rats upon ingestion of [14C]CD14 and exposure to relatively high concentrations of rCD14 led to a reduction in inflammation. This may be beneficial to initial gut colonization in breast-fed newborns. / Alexander Graham Bell NSERC CGS M scholarship. Japan Society for the Promotion of Sciences, Summer in Japan Fellowship. Funded by the Canadian Institutes of Health Research, Institute of Nutrition Metabolism and Diabetes Grant #82816 “Fate and function of breast milk and recombinant human CD14 at mammary and newborn gastrointestinal mucosal epithelia”.