O epitélio gástrico de ratos passa por modificações morfofisiológicas importantes durante o primeiro mês de vida pós-natal e a dieta é um dos principais fatores que influenciam esse desenvolvimento. Ratos receberam dieta com 20% (C) ou 8% (RP) de proteína, durante o período pré e pós-natal. Avaliamos em ratos de 14, 30 e 50 dias os efeitos da restrição protéica sobre proliferação celular no epitélio gástrico, massa do estômago e corpórea, comprimento do intestino e, expressão de ghrelina no estômago e no plasma. O grupo RP apresentou proliferação celular, massa do estômago e corpórea e comprimento do intestino reduzidos em relação ao grupo C. E maior imunomarcação para ghrelina em animais RP com 30 e 50d em comparação ao grupo C. Não houve diferença na imunomarcação entre animais de 14d. Os níveis plasmáticos de ghrelina apresentaram a mesma tendência observada na imunomarcação. O consumo de quantidade adequada de proteína é importante durante o desenvolvimento gástrico de ratos e a ghrelina apresenta resposta hormonal diferente de acordo com a idade do animal. / The gastric epithelium of rats undergoes morphophysiological changes throughout the first month of life and diet is one of the main factors influencing the development. Rats were fed a 20% (NP) or 8% (RP) protein diet throughout the pre- and post-natal life. We analyzed the cell proliferation and observed the body and stomach weight and small intestine length at 14, 30 and 50 days rats. Additionally, we evaluated ghrelin in gastric epithelium and its plasma levels. Cell proliferation in the gastric epithelium, body and stomach weight and small intestine length were reduced in RP animals when compared to NP animals. We observed an increase in the number of labeled cells for ghrelin in 30- and 50-d-old RP rats when compared to the NP group and no difference was found in 14-d-old animals. Plasma ghrelin levels showed the same results observed in immunohistochemical reactions. These results emphasize the importance of diet protein on the development of gastric mucosa and protein restriction seems to differently modulate ghrelin response at different ages.

A obesidade é um dos maiores problemas de saúde pública em todo mundo. A cirurgia Bariátrica é um dos tratamentos mais efetivos para obesidade grave e suas comorbidades. Entre as técnicas de cirurgia bariátrica, a derivação gástrica em y-de-Roux (DGYR) é a mais utilizada. Esse tipo de cirurgia é caracterizada por importantes modificações anatômicas no trato gastrointestinal, como a exclusão de parte do estômago e duodeno, que promovem perda de peso, resolução de comorbidades, como o DM2, e redução de gastos com a saúde. Por outro lado, casos de câncer têm sido observados no estômago excluso de pacientes submetidos à DGYR. Considerando a evidência de câncer de coto gástrico após gastrectomia de Billroth II para úlcera péptica, as mudanças anatômicas após a cirurgia bariátrica podem aumentar o risco carcinogênico. No presente estudo, utilizamos multiplataformas integradas de análises transcriptômicas e metabolômicas combinadas com análises endoscópicas e histológicas para avaliar mudanças precoces no estômago excluso após DGYR. Três meses após a cirurgia, a combinação das diferentes técnicas nos permitiram identificar: refluxo biliar, alterações na mucosa gástrica com a presença de atrofia, inflamação crônica e metaplasia intestinal, que em conjunto promoveram reprogramação gênica e metabólica local, aumentando a expressão de genes associados com o câncer. Pela primeira vez, os dados iniciais revelados pelo presente estudo encorajam pesquisas mais robustas e sugerem o exame endoscópico periódico após a DGYR / Obesity is one of the most important public health conditions worldwide. Bariatric surgery, such as Roux-en-Y gastric by-pass (RYGB), is an effective treatment for severe obesity that results in the improvement and remission of many obesity-related comorbidities as well as providing sustained weight loss, improvement in the quality of life and reduces health spending. On the other hand, cases of cancer have been observed in the excluded stomach, in patients submitted to RYGB. Using the evidence of gastric stump cancer after Billroth II gastrectomy for peptic ulcer, anatomic changes after bariatric surgery may increase carcinogenic risk. We used an integrated transcriptomic and untarget metabolomics multiplataform combined with endoscopic and histological analysis to evaluate early changes in the excluded stomach after RYGB. Three months after surgery, the combination of the different analysis tools allowed us to identify: biliary reflux, mucosal changes with atrophy, chronic inflammation and intestinal metaplasia, which promoted local gene and metabolic reprogramming increasing gene expression associated with cancer. For the first time the initial data revealed by our work encourages more robust research and suggests periodic endoscopic surveillance after RYGB

This work has tried to provide better insight in some pathophysiological factors involved in functional dyspepsia. We have identified several experimental evidences supporting the hypothesis that activation of transducers of wall tension at the level the proximal stomach might be the key to the genesis of at least some of the symptoms. One of the mechanisms by which this activation may be enhanced in patients is the presence of defective accommodation of the proximal stomach in response to a meal. This abnormality was present in a large subgroup of patients and was associated to the presence of early satiety. Pharmacological modulation of the gastric wall tension resulted in concomitant changes in symptom severity, both in health and in functional dyspepsia patients. Special attention has been given to provide the clinician with better tools to investigate his patient, in the perspective of the prescription of a treatment aimed at restoring a defective mechanism. / Ce travail a tenté d'améliorer la compréhension de certains mécanismes physiopathologiques de la dyspepsie fonctionnelle. Nous avons identifié plusieurs arguments expérimentaux soutenant l'hypothèse disant que c'est l'activation de mécanorécepteurs sensibles à la tension pariétale qui est reponsable d'au moins un des symptômes de l'affection. Un des mécanismes par lesquels cette activation peut accrue chez les patients est la présence d'un défaut de l'accommodation gastrique post-prandiale. Cette anomalie a été retrouvée dans une large proportion des patients et est associée à la présence de satiété précoce. Les modifications du tonus gastrique ont résulté dans de modifications concomitantes des symptômes aussi bien chez les volontaires que chez les patients atteints de dyspepsie fonctionnelle.Une attention particulière a été portée sur le développement de nouveaux outils permettant de caractériser les patients dans la perspective de la prescription d'un traitement visant à corriger un mécanisme défectueux.

This work has tried to provide better insight in some pathophysiological factors involved in functional dyspepsia. We have identified several experimental evidences supporting the hypothesis that activation of transducers of wall tension at the level the proximal stomach might be the key to the genesis of at least some of the symptoms. One of the mechanisms by which this activation may be enhanced in patients is the presence of defective accommodation of the proximal stomach in response to a meal. This abnormality was present in a large subgroup of patients and was associated to the presence of early satiety. Pharmacological modulation of the gastric wall tension resulted in concomitant changes in symptom severity, both in health and in functional dyspepsia patients. Special attention has been given to provide the clinician with better tools to investigate his patient, in the perspective of the prescription of a treatment aimed at restoring a defective mechanism. / Ce travail a tenté d'améliorer la compréhension de certains mécanismes physiopathologiques de la dyspepsie fonctionnelle. Nous avons identifié plusieurs arguments expérimentaux soutenant l'hypothèse disant que c'est l'activation de mécanorécepteurs sensibles à la tension pariétale qui est reponsable d'au moins un des symptômes de l'affection. Un des mécanismes par lesquels cette activation peut accrue chez les patients est la présence d'un défaut de l'accommodation gastrique post-prandiale. Cette anomalie a été retrouvée dans une large proportion des patients et est associée à la présence de satiété précoce. Les modifications du tonus gastrique ont résulté dans de modifications concomitantes des symptômes aussi bien chez les volontaires que chez les patients atteints de dyspepsie fonctionnelle.Une attention particulière a été portée sur le développement de nouveaux outils permettant de caractériser les patients dans la perspective de la prescription d'un traitement visant à corriger un mécanisme défectueux.

published_or_final_version / Medicine / Master / Doctor of Medicine

No description available.

In the past, patients with peritoneal surface malignancy were considered incurable and were only offered palliative treatments. However, in a substantial number of patients, disease progression that is isolated to peritoneum may occur. It has been realised that elimination of peritoneal surface tumours may have an impact on the survival of these cancer patients, in whom a prominent cause of death is peritoneal carcinomatosis. The focus of this PhD. thesis is on the combined treatment of cytoreductive surgery and perioperative intrapersonal chemotherapy for diffuse malignant peritoneal mesothelioma, pseudomyxoma peritonei, colorectal peritoneal carcinomatosis and resectable gastric cancer. Section one describes the major principles of management for peritoneal surface malignancy, covering the historical perspectives, the treatment rationales and the learning curve associated with the combined procedure. Section two is devoted to peritoneal mesothelioma, in trying to examine this disease from its clinical, radiologic and histopathologic aspects. A radiologic classification and a histopathologic staging system for this disease are proposed. In section three, the results of the combined treatment for pseudomyxoma peritonei are presented, including a systematic review of the literature, a case series of 50 patients from our Australian centre and a treatment failure analysis of 402 patients from the Washington Cancer Institute. These studies suggest that a disease-free state is important for long-term survival for patients with pseudomyxoma peritonei. In section four, the current evidence on the combined treatment for colorectaI peritoneal carcinomatosis is demonstrated by conducting a systematic review of the literature and survival and perioperative outcome analyses of two separate patient cohorts. These results suggest that the combined treatment is associated with an improved survival, as compared with historical controls. In the last section, a metaanalysis of the randomised controlled trials on adjuvant intraperitoneal chemotherapy for resectable gastric cancer shows that a significant improvement in survival is associated with hyperthermic intraoperative intraperitoneal chemotherapy alone or in combination with early postoperative intraperitoneal chemotherapy.

The development of immune response accountable for the ability to control Cryptosporidium muris TS03 infection was studied using immunocompetent and various types of immunodeficient mouse models. Subsequently the immune response was characterized by analysis of leukocyte infiltration and cytokine production in gastric epithelium. Moreover, the potentiality of immunocompetent mice to develop effective immune response to C. andersoni LI03 infection with consequent protection to consequent infection of the same mice with C. muris TS03 was also studied by monitoring oocysts shedding, leukocyte infiltration of the gastric mucosa and cytokine production in ex vivo cultures of splenocytes.

Sinais clínicos de dispepsia são frequentemente observados em pequenos animais, recomendando-se nesses pacientes a pesquisa de Helicobacter sp., pois esse agente pode estar associado com a ocorrência desses distúrbios. Há poucos relatos de isolamento dessa bactéria em animais no Brasil. Portanto, o objetivo do estudo foi investigar a presença de Helicobacter sp. em pacientes com indicação de endoscopia digestiva alta, atendidos no Hospital de Clínicas Veterinárias da Universidade Federal do Rio Grande do Sul, empregando os métodos diagnósticos mais comumente adotados para sua detecção. Foram colhidos fragmentos de biópsia gástrica de um total de 30 cães e gatos submetidos à endoscopia, que não faziam uso de medicação antimicrobiana e inibidores de bomba de prótons no período mínimo de quatro semanas anteriores à coleta. Os fragmentos foram submetidos ao teste rápido da ureia (URT), colorações histológicas (H&E, Warthin Starry-WS), imunohistoquímica (IHQ), semi-nested PCR, tendo como alvo o gene 16SrRNA e cultura. Foram considerados positivos para Helicobacter sp. os pacientes que apresentaram resultado positivo em pelo menos dois métodos diagnósticos. Em apenas três animais foram detectadas lesões macroscópicas durante a endoscopia e 17 apresentaram lesões histopatológicas na mucosa do estômago. O URT foi positivo em 25 amostras e Helicobacter foi isolado de oito animais. Lâminas coradas por H&E e WS monstraram bactérias com morfologia compatível em 20 e 26 amostras, respectivamente. As técnicas de IHQ e semi-nested PCR resultaram positivas em 27 e 28 amostras, respectivamente. Ao todo, 28 pacientes foram considerados positivos, sendo que IHQ e semi-nested PCR detectaram 96,4% e 100% desses, respectivamente. Dessa forma, conclui-se que ambas as técnicas podem ser recomendadas para a detecção de Helicobacter em pequenos animais. A presença de Helicobacter sp. pode ter causado os sinais gastrintestinais apresentados por cães e gatos incluídos no estudo, porém em muitos pacientes sintomáticos houve presença da bactéria na ausência de lesões microscópicas detectáveis. / Dyspepsia signs are frequently observed in small animals. In these patients, Helicobacter sp. investigation is recommended, since this pathogen may be associated with the occurrence of this kind of disorders. In Brazil there are few reports of Helicobacter sp. detection in small animals. Therefore, the aim of the study was to investigate the presence of Helicobacter sp. by the most common methods adopted for diagnosis in patients referred for upper endoscopy at the Veterinary Hospital of the Federal University of Rio Grande do Sul. Thirty patients, which had not been using any antimicrobial drugs and proton pump inhibitors for at least four weeks prior the endoscopy, were selected for this study. The biopsy fragments were submitted to rapid urea test (URT), histological staining (H&E, Warthin Starry- WS), immunohistochemistry (IHC), semi-nested PCR targeting the 16S rRNA gene and culture. Patients that tested positive in at least two diagnostic methods were considered positive for Helicobacter sp. Macroscopic lesions were detected in only three animals during endoscopy, and seventeen had histopathological lesions in the stomach mucous. The URT resulted in 25 positive samples, while Helicobacter sp. was isolated from only eight patients. Histological staining (H&E and WS) showed bacteria with typical morphology in 20 and 26 samples, respectively. The IHC and semi-nested PCR were positive in 27 and 28 samples, respectively. A total of 28 patients were considered positive; among them IHC and semi-nested PCR resulted positive in 96.4% and 100%, respectively. In conclusion, both techniques can be recommended for Helicobacter sp. detection in small animals. Helicobacter infection may have been the cause of gastrointestinal signs detected in patients; however many symptomatic animals without microscopic lesions were also diagnosed positive.

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No. of bitstreams: 1 stuchi_nmm_dr_sjrp.pdf: 5464428 bytes, checksum: f53c801765e2d85e4f507ebaaebe4625 (MD5) Previous issue date: 2016-05-31 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Apesar do declínio da incidência, o câncer gástrico ocupa ainda a terceira posição em causa de morte por câncer no mundo, tendo como principal fator de risco a bactéria Helicobacter pylori. Esta bactéria pode levar a uma inflamação persistente através da produção de citocinas pró-inflamatórias e de espécies reativas de oxigênio e nitrogênio, estimulando a proliferação celular bem como outros processos envolvidos na carcinogênese. Ainda envolvidos nestes processos tem sido observada a participação de microRNAs, que exercem papel importante na regulação pós-transcricional, influenciando processos fisiológicos normais da célula bem como aqueles ligados às doenças, como por exemplo o câncer gástrico. Alguns miRNAs podem atuar como oncogenes, genes supressores de tumor e biomarcadores para diversas patologias, podendo alvejar genes relacionados com inflamação e câncer como o gene ANXA1 (Anexina-A1). A Anexina-A1 é uma proteína anti-inflamatória e com ação anti-proliferativa, que se liga à receptores do tipo formil peptídeo como por exemplo FPR1 e FPR2, ambos sabidamente relacionados com a progressão de doenças como o câncer. Desta forma o presente trabalho teve como objetivos avaliar a expressão da proteína Anexina A1 e seus receptores FPR1 e FPR2, bem como avaliar a expressão do RNAm da ANXA1 e de miRNAs que possam modular a expressão desse gene (hsa-mir-27a, hsa-mir-196a e hsa-mir-222) em adenocarcinoma gástrico e correlacionar estes resultados com os aspectos clínico-patológicos. Foram avaliadas 31 amostras de adenocarcinoma gástrico, assim como as regiões metaplásica ou normal adjacentes ao tumor. A quantificação relativa (RQ) do RNAm da ANXA1 e miRNAs foi realizada por PCR quantitativo em tempo real utilizando ensaio TaqMan, e a expressão proteica da AnxA1, FPR1 e FPR2 por imuno-histoquímica e análise densitométrica. Em relação à expressão gênica relativa foram observados o aumento da expressão da ANXA1 nos tumores (RQ=1,374; p<0,001), assim como do miR-196a que apresentou aumento de expressão tanto no tecido metaplásico (RQ= 4,784; p=0,0016) e tumoral (RQ=16,99; p< 0,001) em relação ao tecido normal. O miR-27a não se apresentou diferencialmente expresso nos diferentes tecidos e houve diminuição da expressão do miR-222 (RQ=0,687; p=0,01) no tecido tumoral em relação ao tecido normal. Apenas o miR-196a e a ANXA1 apresentaram correlação significantemente inversa (r= -0,55; p=0,003), e este miRNA foi o único que apresentou associação com o sexo feminino, devido aumento de expressão em mulheres. Quando se comparou a expressão relativa aos parâmetros clínico-patológicos e tumorais não foram encontradas diferenças significativas. Quanto à expressão proteica o FPR2 não apresentou marcação no tecido normal, metaplásico e tumoral. Contudo a AnxA1 e FPR1 apresentaram imunomarcação positiva amplamente distribuída no tecido tumoral e positivamente correlacionados tanto no epitélio (r=0,87; p<0,0001) como estroma (r=0,62; p=0,004). Portanto, nossos resultados sugerem que a ANXA1 é modulada pelo miR-196a em câncer gástrico, e evidenciam que tanto a AnxA1 como o FPR1 também estão envolvidos no processo de carcinogênese gástrica. Tais achados podem abrir possibilidades para futuros estudos sobre novos alvos terapêuticos já que AnxA1 e FPR1 são possíveis alvos farmacológicos, e as terapias baseadas em microRNAs tem sido amplamente pesquisadas. / Gastric cancer still ranks third in cause of cancer death worldwide, despite the decline in its incidence, being Helicobacter pylori the main risk factor for this disease. This bacterium can lead to persistent inflammation via the production of proinflammatory cytokines and reactive oxygen and nitrogen species, stimulating cell proliferation and other processes involved in carcinogenesis. Still involved in this process, it has been observed the participation of miRNAs, which play an important role in post- transcriptional regulation, influencing normal physiological processes of the cell as well as those linked to diseases such as gastric cancer. Some miRNAs can act as oncogenes, tumor suppressor genes and biomarkers for various diseases, can targetting genes linked to inflammation and cancer such as ANXA1 gene (Annexin A1). Annexin A1 is an anti-inflammatory protein with anti-proliferative action that binds to formyl peptide receptors such as, FPR1 and FPR2, both known to be related to the progression of diseases such as cancer. Thus, the present study aimed to evaluate the expression of Annexin A1, FPR1 and FPR2 receptors, and the expression of mRNA ANXA1 and miRNAs (hsa-mir-27a, hsa-mir-196a and hsa-miR 222) in gastric adenocarcinoma, also correlate these results to the clinicopathological features. 31 adenocarcinoma samples were evaluated, as well as normal or metaplastic region adjacent to the tumor. Relative quantification (RQ) of miRNA and mRNA ANXA1 was evaluated by TaqMan assay and protein expression AnxA1, FPR1 and FPR2 by immunohistochemistry and densitometry analysis. Regarding the relative expression the following results were observed, increased expression in tumors of ANXA1 (RQ = 1.374; p < 0.001) and miR- 196a that showed increased expression it he metaplastic tissue (RQ = 4.784; p = 0.0016) and tumor (RQ = 16.99; p < 0.001) compared to normal tissue. The miR- 27a did not appear differentially expressed in different tissues and there was a decrease of miR -222 expression (RQ = 0.687; p = 0.01) in tumor tissue compared to normal tissue. Only the miR-196a and ANXA1 presented a significant inverse correlation (r = -0.55; p = 0.003), and this miRNA was the only one that showed association to female gender, due to increased expression in women when comparing the relative expression to clinicopathological parameters and tumor features. The FPR2 was not expressed in normal, metaplasic and tumor tissue. However, the AnxA1 and FPR1 were widely distributed positive immunostaining in tumor tissue and positively correlated both in the epithelium (r = 0.87 ; p < 0.0001) and stromal (r = 0.62 ; p = 0.004). Thus, our results suggest that ANXA1 is modulated by miR-196a in gastric cancer, and evidencing that both AnxA1 and FPR1 are also involved in gastric carcinogenesis process. These data open the possibility for future studies on new therapeutic targets since AnxA1 and FPR1 are adjustable pharmacological targets, and microRNAs ` therapies have been widely researched. / FAPESP: 2012/15036-8 / CNPq: 474776/2013-1