Homeodynamic space (HDS) shrinks as vulnerability increases with aging and repeated damage to the cells. HDS is lost in alcoholic pancreatitis patients due to overconsumption of alcohol, smoking, and high fat diets. Etiologically relevant animal models for study of chronic pancreatitis (CP) are needed. In order to begin filling this gap a central purpose of this dissertation research was to examine relationships between the alcohol and high fat diet (AHF) and pancreatitis with attention to hypersensitivity and anxiety-like behaviors. The AHF diet induced pancreatitis described here etiologically mimics human risk factors of AHF consumption for advancement to alcoholic CP. In this study one group of mice was fed long term with a diet of high fat and alcohol for comparison with a group fed normal chow. Mice consumed a liquid diet containing 6% alcohol and a high fat supplement ad libitum over a period of five months. Each group was evaluated for heat and mechanical hypersensitivity, and histology indicative of CP. The association of pancreatitis pathology with anxiety has been understudied. Anxiety, like pain, is useful as a transient state but when anxiety is prolonged it is termed a disorder. Anxiety is often comorbid with pain and depression. Therefore, it is important to determine anxiety in mice with CP histology. This model was characterized for the interaction of pancreatitis histology, as well as persisting pain-, anxiety-, and fear-like behaviors. The AHF diet mice developed hypersensitivity, demonstrated anxiety-like behaviors, and showed concurrent histology consistent with CP. Nontransgenic mouse models where pancreatitis is induced only by a combination of ad libitum liquid food with added alcohol and lard supplementation do not currently exist, nor has an in-depth study of anxiety-like behaviors been conducted in this mouse model. This dissertation research addresses this knowledge gap.

A perfuração gástrica por endoscopia é a consequência de alguns de seus procedimentos, e atualmente, com o advento das cirurgias endoscópicas translumenais por orifícios naturais, um meio de manipulação dos órgãos abdominais. Esse é o motivo pelo qual os endoscopistas estão procurando um reparo endoscópico seguro. O objetivo foi avaliar a factibilidade e os resultados do fechamento da abertura gástrica similar àquelas realizadas nos procedimentos cirúrgicos endoscópicos translumenais por orifícios naturais utilizando-se o T-Tag associado à câmara plástica protetora. Sob anestesia geral, dez porcos Landrace foram submetidos a uma perfuração gástrica calibrada em 18 mm de diâmetro. A abertura foi fechada pelo novo método apresentado, composto de fios cirúrgicos conectados em uma âncora metálica (T-Tag) posicionados pela parede gástrica através de uma agulha. Uma câmara protetora plástica foi adaptada à extremidade distal do endoscópio para proteger os órgãos abdominais adjacentes da punção da agulha fora do estômago. Seis dispositivos T-Tag foram posicionados na maioria dos casos e os fios atados com um apertador de nó metálico endoscópico formando três pontos de sutura. O teste de vazamento foi realizado com uma pinça endoscópica e distensão da câmara gástrica com ar. Os animais receberam líquidos no mesmo dia do procedimento. Uma dose de antibiótico diária por dois dias foi administrada. Nenhuma complicação foi detectada no período pós-operatório. Um mês depois, a endoscopia revelou a presença de cicatriz em todos os animais, e a maioria apresentava materiais da sutura aderidos à superfície mucosa da região. A região do antro gástrico apresentava poucas aderências identificadas na laparotomia realizada no mesmo momento. O reparo endoscópico utilizando o T-Tag e a câmara protetora plástica é factível, fácil de ser realizada e seguro. São necessários estudos adicionais para mostrar o real valor desse tipo de procedimento / The endoscopic gastric perforation is a consequence of some endoscopic procedures and now a way to manage abdominal organs. This is the reason why endoscopists are studying a safe endoscopic repair. The objective was to evaluate feasibility and results of the gastric opening closure similar to those performed in natural orifice translumenal endoscopic surgery procedures using T-Tag associated with the plastic protection chamber. Ten Landrace pigs underwent a gastric perforation of 1.8 cm in diameter under general anesthesia. The opening was repaired with stitch assembled in a T-Tag anchor placed through the gastric wall with a needle. A plastic transparent chamber, adapted to the endoscope tip protected the abdominal organs from the needle puncture outside the stomach. Six T-Tags were placed in most cases and the stitches were tied with a metallic tie-knot, forming three sutures. The leakage test was performed with a forceps and by air distention. The animals received liquids in the same operative day. One daily shot antibiotic during two days was used. No complication was detected in the postoperative course. One month later the endoscopy revealed a scar in all animals, and the majority of these with suture material. The antral anterior gastric wall was clear with few adhesions in the laparotomy performed in the same time. The endoscopic repair using T-Tag and a protector chamber is feasible, easy to perform and safe. Further studies are needed to show the real value of this kind of procedure

Hepatitis C Virus (HCV) is the primary cause of liver transplantation due to its chronic nature in up to eighty percent of infected cases. Around 3 percent of the world’s population is infected with HCV. Treatment for HCV is a combined Ribavirin and interferon-α (IFN-α) therapy effective in only fifty to eighty percent of patients depending on HCV genotype. The growing health concern with this disease is the lack of a cure despite liver transplantation. HCV targets hepatocytes, liver cells, but is not cytolytic. HCV has been shown to induce end stage liver disease through sustained inflammation from the host’s immune system in the liver. One of the key dilemmas in HCV research and the search for fully effective treatments or vaccines is the lack of animal models. HCV infectivity and disease is limited to primates, most specifically to humans, which cannot be fully replicated in any other living being. The mechanisms for HCV evasion or activation of the immune system are complex, many and discoveries within this field are crucial to overcoming this destructive hepatic infection. Toll-like receptors (TLR) are cellular activators of the innate immune system that have been a target of HCV. Activated TLRs trigger both the inflammatory and anti-viral pathways to produce inflammatory cytokines and interferons. HCV proteins have been reported to activate a number of TLRs in a variety of cell types. In order to identify possible targets of HCV within the TLR family, we first characterized TLR presence and function in both human hepatic carcinoma cell lines and purified primary human hepatocytes. RNA from TLRs 1-10 was observed to varying degrees in both the hepatoma cell lines and the primary hepatocytes. We show the extracellular and/or intracellular presence of TLR2, TLR1, TLR3 and TLR7 proteins in hepatoma cell lines. TLR3 and TLR7 are located within the endosome and recognize viral RNA products. We recently reported that TLR2-mediated innate immune signaling pathways are activated by HCV core and NS3 proteins. TLR2 activation requires homo- or heterodimerization with either TLR1 or TLR6. We show NF-κB activation in hepatoma cells by TLR2/1, TLR2/6 ligand and HCV protein stimulation. In primary hepatocytes, HCV proteins induced both IL-8 and IL-6 production. We also show that primary hepatocytes initiate a Type 1 IFN response in addition to IL-8 and IL-6 production upon stimulation with a TLR7/8 ligand. Human hepatoma and primary hepatocytes are responsive to TLR2, TLR1, TLR6, TLR7/8 ligands and HCV proteins. Activation of these TLRs may contribute to the inflammatory mediated destruction caused by HCV or could be targets of HCV contributing to its immune evasion. We found previously that hepatoma cells and primary hepatocytes are responsive to TLR2 ligands and HCV proteins. We also reported that TLR2 is activated by HCV proteins. Here we aimed to determine whether TLR2 coreceptors participated in cellular activation by HCV core or NS3 proteins. By designing siRNAs targeted to TLR2, TLR1 and TLR6, we showed that knockdown of each of these receptors impairs pro- and anti-inflammatory cytokine activation by TLR-specific ligands as well as by HCV core and NS3 proteins in Human Embryonic Kidney cells (HEK/TLR2) and in primary human macrophages. We found that HCV core and NS3 proteins induced TNF-α and IL-10 production in human monocyte-derived macrophages, which was impaired by TLR2, TLR1 and TLR6 knockdown. Contrary to human data, results from TLR2, TLR1 or TLR6 knockout mice indicated that the absence of TLR2 and its coreceptor TLR6, but not TLR1, prevented the HCV core and NS3 protein-induced peritoneal macrophage activation. TLR2 may utilize both TLR1 and TLR6 coreceptors for HCV core- and NS3-mediated activation of macrophages and innate immunity in humans. These results imply that multiple pattern recognition receptors could participate in cellular activation by HCV proteins contributing to inflammatory disease. Two critical factors in chronic HCV infection are inflammatory disease and immune evasion. We have demonstrated that TLR2 and its co-receptors play a role in inflammatory-mediated induction via HCV NS3 and core administration. It has recently been shown that HCV targets the TLR3 pathway to aid in immune evasion. TLR3 is only one of four viral recognition receptors located within the endosome and it is plausible that HCV may target others. We hypothesized that HCV infection may interfere with the expression and function of TLR7, a sensor of single stranded RNA. Investigating any effect on TLR7 by HCV may reveal a new mechanism for HCV immune evasion. Low levels of both TLR7 mRNA and protein were measured in HCV replicating cells compared to control cells while reducing HCV infection with either IFNα or restrictive culture conditions restored the decreased TLR7 expression. Downstream of the TLR7 pathway, an increased baseline IRF7 nuclear translocation was observed in HCV replicating cells compared to controls. Stimulation with a TLR7 ligand, R837, resulted in significant IRF7 nuclear translocation in control cells. In contrast, HCV replicating cells showed impaired IRF7 activation. Use of RNA polymerase inhibitors on hepatoma cells, control and HCV replicating, revealed a shorter TLR7 half life in HCV replicating cells compared to control cells which was not seen in TLR5 mRNA. These data suggest that reduced TLR7 expression, due to RNA instability, directly correlates with HCV replication and results in impaired TLR7-induced IRF7-mediated cell activation. In conclusion, Hepatitis C Virus manipulates specific Toll-like receptors’ expression and their signaling pathways to induce cytokine production. HCV utilizes surface receptors TLR2 and its co-receptors which once activated could contribute to inflammatory disease by production of inflammatory cytokines and possibly immune evasion. HCV down-regulates TLR7, a viral recognition receptor, by decreasing mRNA stability which could facilitate evasion of host immune surveillance.

A perfuração gástrica por endoscopia é a consequência de alguns de seus procedimentos, e atualmente, com o advento das cirurgias endoscópicas translumenais por orifícios naturais, um meio de manipulação dos órgãos abdominais. Esse é o motivo pelo qual os endoscopistas estão procurando um reparo endoscópico seguro. O objetivo foi avaliar a factibilidade e os resultados do fechamento da abertura gástrica similar àquelas realizadas nos procedimentos cirúrgicos endoscópicos translumenais por orifícios naturais utilizando-se o T-Tag associado à câmara plástica protetora. Sob anestesia geral, dez porcos Landrace foram submetidos a uma perfuração gástrica calibrada em 18 mm de diâmetro. A abertura foi fechada pelo novo método apresentado, composto de fios cirúrgicos conectados em uma âncora metálica (T-Tag) posicionados pela parede gástrica através de uma agulha. Uma câmara protetora plástica foi adaptada à extremidade distal do endoscópio para proteger os órgãos abdominais adjacentes da punção da agulha fora do estômago. Seis dispositivos T-Tag foram posicionados na maioria dos casos e os fios atados com um apertador de nó metálico endoscópico formando três pontos de sutura. O teste de vazamento foi realizado com uma pinça endoscópica e distensão da câmara gástrica com ar. Os animais receberam líquidos no mesmo dia do procedimento. Uma dose de antibiótico diária por dois dias foi administrada. Nenhuma complicação foi detectada no período pós-operatório. Um mês depois, a endoscopia revelou a presença de cicatriz em todos os animais, e a maioria apresentava materiais da sutura aderidos à superfície mucosa da região. A região do antro gástrico apresentava poucas aderências identificadas na laparotomia realizada no mesmo momento. O reparo endoscópico utilizando o T-Tag e a câmara protetora plástica é factível, fácil de ser realizada e seguro. São necessários estudos adicionais para mostrar o real valor desse tipo de procedimento / The endoscopic gastric perforation is a consequence of some endoscopic procedures and now a way to manage abdominal organs. This is the reason why endoscopists are studying a safe endoscopic repair. The objective was to evaluate feasibility and results of the gastric opening closure similar to those performed in natural orifice translumenal endoscopic surgery procedures using T-Tag associated with the plastic protection chamber. Ten Landrace pigs underwent a gastric perforation of 1.8 cm in diameter under general anesthesia. The opening was repaired with stitch assembled in a T-Tag anchor placed through the gastric wall with a needle. A plastic transparent chamber, adapted to the endoscope tip protected the abdominal organs from the needle puncture outside the stomach. Six T-Tags were placed in most cases and the stitches were tied with a metallic tie-knot, forming three sutures. The leakage test was performed with a forceps and by air distention. The animals received liquids in the same operative day. One daily shot antibiotic during two days was used. No complication was detected in the postoperative course. One month later the endoscopy revealed a scar in all animals, and the majority of these with suture material. The antral anterior gastric wall was clear with few adhesions in the laparotomy performed in the same time. The endoscopic repair using T-Tag and a protector chamber is feasible, easy to perform and safe. Further studies are needed to show the real value of this kind of procedure

Autism spectrum disorders (ASD) encompass neurodevelopment disorders characterized by atypical patterns of development that impact multiple areas of functioning beginning in early childhood. The etiology of ASD is unknown and there are currently no preventative treatment options. Gastrointestinal symptoms are commonly associated comorbidities. The microbiota-gut-brain axis is a multidirectional communication chain that connects the central and enteric nervous system that relates brain function to peripheral intestinal functions. Changes within this axis have been postulated in ASD. For example, the “leaky gut theory” proposes that chronic inflammation is linked to alterations in the bacterial profiles of the gut microbiome and subsequent shifts in the amount and type of short-chain fatty acids produced can affect downstream neuronal development. Short-chain fatty acids are signaling molecules produced by bacteria that can trigger nerve afferents in the gut. Dysbiosis causes altered signaling patterns that can be identified by altered intestinal morphology. In this study, C57BL/6J control mice and three mouse models of social behavioral deficits were used to investigate markers of intestinal pathophysiology. Fecal and intestinal samples were collected from adult wild type control mice and the social deficit groups of BTBR genetic knockout mice, C57BL/6J mice injected with valproic acid, and C57BL/6J mice injected with polyinosinic–polycytidylic acid. Short-chain fatty acid profiles that included acetic, propionic, isobutyric, butyric, isovaleric, and valeric acids were obtained from fecal samples to determine differences between the models and control mice. The profiles of the BTBR genetic knockout and valproic acid models were found to be significantly different from control mice. Additionally, postmortem intestinal ileum samples underwent hematoxylin and eosin identification procedures to determine the thickness of the tunica muscularis and tunica mucosa. The thickness of the tunica muscularis was reduced in the valproic acid group compared to the wild type control mice in early stages of development (p=0.0279). This research may illuminate developmental cues that attribute to autism spectrum disorders and may provide markers to assess future therapeutic treatments.

No trato digestório a colite ulcerativa apresenta necrose no intestino como processos patofisiológicos. Este projeto visou estudar neurônios com códigos químicos do sistema nervoso entérico (SNE) e a morfologia estrutural do intestino grosso de animais com colite ulcerativa. Grupos: a) Colite: injetados com TNBS, b) PBS: injetados com PBS e c) controle. Os tecidos foram preparados por métodos imunohistoquímicos de duplas marcações do receptor P2X7 com NOS, ChAT, Calb, Calr, anti-HuC/D (pan-neuronal) e S100 (células glias). No grupo Colite, no plexo mioentérico, o receptor P2X7 estava diminuído. No tecido lesado apresentou aumento de neutrófilos e da lâmina própria, alteração de colágeno e destruição do epitélio e células caliciformes. Reduziram colocalizações de neurônios com receptor no plexo mioentérico e aumento no plexo submucoso. Houve reduções nas densidades e áreas dos neurônios no SNE. Conclui-se que a colite afetou os neurônios entéricos e células gliais, causou alterações morfológicas, sendo assim, pode afetar motilidade intestinal. / In the digestive tract ulcerative colitis have a bowel necrosis as pathophysiological processes. This project aimed to study neurons with their respective chemicals codes the enteric nervous system (ENS) as well as structural morphology of the distal colon of animals with ulcerative colitis. Groups: a) colitis: TNBS injected, b) PBS: PBS injected and c) control. The tissues were prepared by immunohistochemical methods for double marking with P2X7 receptor, ChAT, Calb, Calr, anti-HUC / D (pan-neuronal) and S100 (glial cells). In the colitis group, the myenteric plexus, the P2X7 receptor decreased. In the injured tissue showed increased neutrophils, alteration and destruction of collagen and epithelial goblet cells. There were reduced colocalizations of neurons with receptor in myenteric plexus and increase in submucosal plexus. There were reductions in the densities and areas neurons of ENS. Concluded that colitis affected enteric neurons and glial cells, causing morphological changes and could be affect intestinal motility.

Introdução: Muitos pacientes desenvolvem recidiva de câncer colorretal (CCR), que possui importância epidemiológica devido ao diagnóstico tardio e à dificuldade de acessibilidade à assistência de saúde. Objetivos: analisar as evidências científicas sobre a deiscência de anastomose intestinal em cirurgias por laparotomia e laparoscopia em pacientes com CCR, a recidiva de câncer colorretal e a assistência de enfermagem para pacientes com recidiva de CCR em tratamento cirúrgico; e realizar proposição de assistência de enfermagem perioperatória para esta clientela, com base nestas evidências científicas. Materiais e método: trata-se de revisão integrativa (RI), fundamentada na Prática Baseada em Evidências, realizada com as etapas de elaboração da pergunta e definição das estratégias de busca; elaboração de critérios de inclusão e exclusão; utilização de um instrumento para a coleta de dados da amostra; análise crítica dos estudos com avaliação do rigor e das características da amostra; síntese e interpretação dos resultados; e apresentação da RI. Para a busca da literatura científica utilizou-se os descritores colorectal neoplasms, laparoscopy, laparotmy e anastomoticleak; recurrence e colorectal neoplasm; nas base de dados Cumulative Index to Nursing and Allied Health Literature (CINAHL), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), Medical Literature Analysis and Retrieval System Online (MEDLINE), Biblioteca virtual COCHRANE, Índice Bibliográfico Español de Ciencias de la Salud (IBECS) e biblioteca virtual National Library of Medicine National Institutes of Health (PUBMED), mediante os critérios de inclusão e de exclusão, com seleção de oito e quatro artigos científicos, respectivamente; e com os descritores colorectal neoplasms, recurrence e nursing, seis artigos científicos. Resultados e discussão: a ocorrência de deiscência de anastomose, foi analisada em relação ao uso de novas técnicas cirúrgicas e maior densidade tecnológica para as terapêuticas adjuvantes, farmacológicas e os exames diagnósticos, que comprometem a mortalidade e sobrevida desta clientela. Em geral, os pacientes apresentam estadiamento avançado da doença, com realização de cirurgia tradicional pela condição clínica associada à comorbidades, maior risco cirúrgico e ampla ressecção intestinal, o que comprometem a sua recuperação fisiológica e psicossocial, além de maior possibilidade de recidiva ou metástases. Para a assistência à saúde desta clientela há necessidade de equipe multidisciplinar, priorizando-se a complexidade clínica, utilização de maior densidade tecnológica e seguimento especializado ao longo da sobrevivência. Há escassez de produção científica, sobre aspectos clínicos, terapêuticos e preventivos, principalmente da enfermagem. Conclusão: O enfermeiro deve ter domínio sobre fisiopatologia, terapêuticas e suas consequências, além de prognósticos e aspectos preventivos do CCR, para prestar assistência de enfermagem perioperatória para pacientes com CCR e familiares, com planejamento de intervenções procedimentais, educativas e psicossociais / Background: Many patients develop colorectal cancer recurrence (CCR), with epidemiological importance due to late diagnosis and the difficult access to health care. Objectives: To analyse scientific evidence on bowel anastomosis dehiscence in laparotomy and laparoscopy surgeries in patients with CCR, the colorectal cancer recurrence and the nursing care for patients with CCR relapse undergoing surgical treatment; and make the perioperative nursing care proposal to these patients, based on the scientific evidences cited. Method and materials: An integrated review (IR), grounded on the Evidence-Based Practice, developed in the following phases: creation of the question and definition of search strategies; creation of inclusion and exclusion criteria; the use of an instrument to collect sample data; critical analysis of the study and assessment of the rigor and characteristics of the sample; summary and interpretation of the results; and IR presentation. In the search for scientific literature, the descriptors colorectal neoplasms, laparoscopy, laparotomy and anastomotic leak; recurrence and colorectal neoplasm were used on the following databases: Cumulative Index to Nursing and Allied Health Literature (CINAHL), LatinAmerican and Caribbean Literatures on health Science (LILACS), Medical Literature Analysis and Retrieval System Online (MEDLINE), COCHRANE virtual library, Español de Ciencias de la Salud (IBECS) bibliographic index and National Library of Medicine National Institutes of Health (PUBMED) virtual library, using the inclusion and exclusion criteria, selecting 8 and 4 scientific articles, respectively; and 6 scientific articles with the descriptors colorectal neoplasms, recurrence e nursing. Results and discussion: the occurrence of anastomosis dehiscence was analysed in relation to the use of new surgical techniques and greater technological density for the therapeutics adjuvants, pharmacological and the diagnostic tests that affect the mortality and the survival of such patients. In general, patients present advanced staging of the disease with the practice of traditional surgery due to clinic condition associated to comorbidities, greater surgical risk and extensive intestinal resection, which impair their physiological and psychosocial recovery, in addition to a greater possibility of recurrence or metastasis. For the health care of such patients, it is necessary a multidisciplinary team, prioritising the clinical complexity, the use of higher technological density and specialised assistance throughout the survival. There is a shortage of scientific production on clinical, therapeutic and preventive aspects, particularly in nursing. Conclusion: Nurses shall master pathophysiology, therapeutics and their consequences, as well as the prognosis and preventive aspects of CCR in order to provide perioperative nursing care for patients with CCR and their family, with procedural, educational and psychosocial interventions

Patients with liver disease have an increased risk for malnutrition because of side effects of the disease. The Nutrition Focused Physical Exam (NFPE) was developed for nutrition professionals to aid physicians in a nutrition-based diagnosis of malnutrition. The purpose of this study was to examine the NFPE for its validity in liver disease patients being evaluated for transplant. In addition, the NFPE was used to assess incidence and severity of malnutrition in end stage liver disease patients and compare these results to already developed malnutrition tools such as the Patient Generated-Subjective Global Assessment (PG-SGA), Triceps Skinfolds (TSF), Mid-Arm Circumference (MAC), Lumbar Index, and Total Psoas Muscle Area (TPA). The NFPE was found to be highly correlated with PG-SGA results. There was a weak correlation between the NFPE and the TSF, MAC, and Lumbar Index/TPA, except when comparing the bottom 25% quartile of the Lumbar Index to severe malnutrition using the NFPE. This resulted in a moderate correlation. The odds-ratio for hospital admission based on malnutrition and severe malnutrition were both extremely high (14.571, 18.857 respectively). These preliminary results reinforce the significance of the NFPE and the need for additional studies using this tool.

Crohn’s disease (CD) affects about 780,000 people in the United States alone, and it is estimated that 6-15 per 100,000 persons will receive a diagnosis of this disease each year. There currently is no cure for Crohn’s disease, and available medical therapies simply serve to alleviate the inflammation. This does not help treat fibrostenosis that Crohn’s disease patients may develop, which can only be treated surgically. Finding alternatives to treat CD requires an understanding of mechanisms at the biochemical level. In this thesis, we attempted to gain a better understanding of certain pathways found to be active in Crohn’s disease-affected ileal smooth muscle cells. We found an upregulation of the ER stress pathway via expression of its surrogate, the GRP78 protein. We also showed evidence that the phosphoinositide 3-kinase (PI3K) pathway, a key proliferative pathway, is linked to ER stress in these cells, and is an upstream driving force of the ER stress response. Further research on the link between the PI3K and ER stress pathways needs to be conducted, and can potentially serve as a target for therapeutics to help reduce proliferation in fibrostenotic Crohn’s disease-affected ileal smooth muscle cells.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.