O objetivo do presente trabalho foi diagnosticar a doença inflamatória intestinal crônica em cães acometidos por sinais gastrointestinais bem como avaliar a contribuição de diferentes parâmetros utilizados no diagnóstico desta afecção. Para tanto, 20 cães apresentando sinais compatíveis com a doença foram incluídos no estudo (grupo afetado) e comparados com 20 animais saudáveis (grupo controle). Foram atribuídos escores clínicos e realizadas avaliações endoscópicas e histopatológicas dos animais do grupo afetado. Amostras de sangue e fezes de ambos os grupos foram coletadas, armazenadas a -80°C e, posteriormente, utilizadas para a mensuração de TNF-alpha, proteína C-reativa, calprotectina, proteína S100A12, Folato e Cobalamina. A mediana dos valores de albumina sérica foi 3,17 e apenas um animal apresentou hipoalbuminemia. Todas as avaliações endoscópicas mostraram algum grau de alteração nos parâmetros avaliados, sendo que o edema e a hiperemia foram os mais observados. Quanto à avaliação histopatológica, processos do tipo linfocítico plasmocítico foram os mais diagnosticados, presentes em 12 dos 17 acometimentos gástricos, nove dos 15 acometimentos duodenais e nove de 11 acometimentos colônicos. Quando se analisam os escores atribuídos aos parâmetros gástricos, 67,3% das avaliações foram consideradas normais, enquanto que 27,4% foram considerados com alterações leves. Para os fragmentos duodenais, 55,3% das avaliações demonstraram tecidos normais enquanto que 36,0% foram considerados com alterações leves. No que se refere ao cólon, apenas 38,6% dos parâmetros avaliados foram normais enquanto que 45,4% receberam avaliações de alterações leves. As medianas dos escores atribuídos para estômago, duodeno e cólon foram, respectivamente, 3; 3 e 6. A comparação entre valores de proteína C-reativa entre grupo afetado e controle apresentou diferença estatisticamente significante. Esta diferença também foi significante quando se comparou o grupo controle aos subgrupos afetado I e afetado II. Embora valores séricos de calprotectina não foram estatisticamente significantes, a mensuração fecal desta proteína foi mostrou diferença significante entre os grupos. Em relação mensuração sanguínea da proteína S100A12, os valores de medianas de ambos os grupos foram próximos (195,90 µg/L e 195,55 µg/L para grupos afetado e controle, respectivamente) e a comparação entre estes valores não foi estatisticamente significante. Esta diferença também não foi significante na comparação dos valores fecais. Mensurações de folato e cobalamina não se mostraram estatisticamente significante na comparação entre animais afetados e sadios. Observaram-se correlações entre valores de proteína C-reativa e escore clínico, proteína C reativa e escore histopatológico do cólon, albumina e calprotectina fecal, albumina e S100A12 fecal, valores séricos de S100A12 e calprotectina, valores fecais de S100A12 e calprotectina. / The purpose of this study was to diagnose inflammatory bowel disease in dogs with gastrointestinal signs as well as to evaluate the contribution of different parameters used in the diagnosis of this disease. Twenty dogs with compatible signs of the disease were included in the study (affected group) and compared with 20 healthy animals (control group). Clinical scores were assigned and endoscopic and histopathological analysis were performed in affected animals. Samples of blood and feces were collected from both groups, stored at -80°C and then used to measure TNF-alpha, C-reactive protein, calprotectin, S100A12 protein, folate and cobalamin. Seric albumin median level was 3.17g/dL and only one animal had hypoalbuminemia. All endoscopic evaluations showed some degree of change in the assessed parameters, and hyperemia and edema were the most frequently ones observed. In the histopathological evaluation, lymphocytic plasmocytic process was the most frequently diagnosed (12 out 17 gastric conditions, nine out 15 duodenal conditions and nine out 11 colonic conditions). When analyzing the scores assigned to the gastric parameters, 67.3% of the evaluations were considered normal, while 27.4% presented slight alterations. For duodenal fragments, 55.3% of the evaluations showed normal tissue, while 36.0% were considered slightly altered. Regarding the colon, only 38.6% of these parameters were normal, whereas 45.4% of the evaluations presented slight changes. The median of the scores assigned to the stomach, duodenum and colon were 3, 3 and 6, respectively. The comparison of the Creactive protein values between affected and control groups showed statistical difference. This difference was also significant when comparing the control group to affected I and affected II subgroups. Although seric values of calprotectin werent statistically significant, fecal data statistically differed between the groups. Regarding seric protein S100A12 measurement, the median values of both groups were similar (195.90 mg/L and 195.55 mg/L for the affected and control groups, respectively) and the difference between these values was not statistically significant. The comparison between fecal S100A12 protein values didnt showed a significant difference. Folate and cobalamin measurements showed no statistical difference when comparing healthy and affected animals. A direct positive correlation was found between C-reactive protein and clinical score. C-reactive protein and histopathological score of colon, albumin and fecal calprotectin, albumin and fecal protein S100A12, seric protein S100A12 and seric calprotetin and fecal protein S100A12 and fecal calprotectin were correlated.

As doenças inflamatórias intestinais (DII) - doença de Crohn e retocolite ulcerativa, são condições crônicas que, na maioria dos casos, exigem controle com terapia medicamentosa. A atenção farmacêutica (AF), definida como a provisão responsável do tratamento farmacológico, com o propósito de alcançar resultados concretos que melhorem a qualidade de vida do paciente, constitui nova forma de cuidado ao paciente, que necessita ser mais extensivamente avaliada. Este estudo teve o objetivo de avaliar a contribuição da atenção farmacêutica ao tratamento clínico de pacientes com DII em acompanhamento ambulatorial em hospital terciário. Ao longo de um ano, foi avaliado um grupo que recebeu a atenção farmacêutica (GAF; N=18) e um grupo controle (GC; N=17) não submetido aos procedimentos da AF. Os resultados da contribuição da AF foram avaliados pela comparação de diferentes variáveis entre os grupos, que foram obtidas na primeira entrevista - T(0), aos seis - T(6) e 12 - T(12) meses do estudo. Quanto aos aspectos clínicos, houve redução significativa dos índices de atividade clínica de T(6) para T(12) no GAF (mediana; variação: 2,20; 0,99 3,77 versus 1,90; 0,99 3,77; p=0,02), o que não ocorreu no GC (1,69; 0,99 3,77 versus 1,69; 0,99 3,48). No GAF, houve aumento significativo do percentual de pacientes mais aderentes ao tratamento medicamentoso (27,8% versus 72,2%; p<0,05), quando da avaliação por meio do teste de Morisky, mas não foram observadas diferenças (72,2% versus 88,9%) na adesão avaliada pelo cotejo entre medicamentos utilizados e prescrições registradas. Em ambas as formas de avaliação da adesão, tanto pelo teste de Morisky (41,2% versus 41,2%), quanto pelo confronto das medicações utilizadas e prescrições registradas (88,2% versus 82,4%), não foram observadas alterações no GC. Houve aumento significativo dos índices de conhecimento do paciente sobre o tratamento no GAF entre T(0) e T(12) (mediana; variação: 80%; 40% 100% versus 100%; 100% 100%; p0,0001), o que não ocorreu no GC (80%; 0 100% versus 80%; 60% 100%). No que se refere à qualidade de vida, avaliada pelo instrumento SF36, houve diferenças estatisticamente significativas nos dois grupos apenas no domínio de saúde mental. No GAF, houve elevação dos escores deste domínio entre T(0) e T(12) (54,0 versus 66,0; p=0,04), o que, também ocorreu no GC (60,0 versus 68,0; p=0,01). Porém, no GAF, esta mudança ocorreu mais precocemente, de T(0) para T(6) (54,0 versus 66,0; p<0,01). A AF possibilitou a identificação, em média, de 3,8 problemas relacionados ao medicamento por paciente, que em sua maioria foram resolvidos, com intervenções predominantemente focadas em orientações aos pacientes. Os pacientes do GAF, ao término do estudo, apresentaram alto grau de satisfação com a AF. Os resultados obtidos permitem concluir que a introdução de um programa de atenção farmacêutica a pacientes ambulatoriais com DII seguidos em hospital terciário trouxe contribuição positiva, proporcionando benefícios mensuráveis aos pacientes. / Inflammatory bowel diseases (IBD) Crohns disease and ulcerative colitis are chronic conditions which are usually controlled with drug therapy. Pharmaceutical care (PC), defined as the responsible provision of drug therapy for the purpose of achieving definite outcomes that improve patients quality of life, is a new patient care modality, which needs to be more extensively evaluated. This study aimed at assessing the contribution of pharmaceutical care to the clinical treatment of outpatients with IBD assisted at a reference hospital. During one year, a group receiving pharmaceutical care (PCG; N=18) and a control group (CG; N=17), which did not undergo PC procedures, were evaluated. Results of PC contribution were assessed by comparing the two groups regarding different variables obtained in the first interview at - T(0), at six - T(6) and 12 - T(12) months of study. Regarding the clinical aspects, there was a significant decrease of clinical activity indexes from T(6) to T(12) in the PCG (median; range: 2.20; 0.99 3.77 versus 1.90; 0.99 3.77; p=0.22), but not in the CG (1.69; 0.99 3.77 versus 1.69; 0.99 3.48). In the PCG, there was a significant increase in the percentage of patients who were more compliant to drug treatment (27.8 % versus 72.2 %; p<0.05) as assessed using the Morisky scale; however, no differences in compliance rates were observed (72.2 % versus 88.9 %) by comparing drugs taken with registered prescriptions. In the CG, no differences were observed in none of the compliance assessment methods, neither by the Morisky scale (41.2% versus 41.2%), nor by comparing drugs taken with registered prescriptions (88.2% versus 81.2%). There was a significant increase in the values of an index for patients knowledge about the treatment in the PCG between T(0) and T(12) (median; range: 80%; 40 100 versus 100%; 100 100; p0,0001), but not in the CG (80%; 0 100 versus 80%; 60 100). With respect to quality of life, assessed by the SF36 scale, there were statistically significant differences in both groups only in the mental health domain. There was an increase in scores for this domain between T(0) and T(12) in both PCG (54.0 versus 66.0; p=0,04), and CG (60.0 versus 68.0; p=0,01). However, PCG had this increasing scores earlier, between T(0) and T(12) (54.0 versus 74.0; p<0,01). PC enabled the identification of a number of drug-related problems per patient (mean = 3.8), which were mostly solved by interventions predominantly focused on patient orientation. At the end of the study, patients in the PC group showed a high degree of satisfaction with the intervention. The achieved results allow concluding that the implementation of a pharmaceutical care program to outpatients with IBD followed at a tertiary hospital gave a positive contribution, providing measurable benefits to patients.

Chronic pelvic pain (CPP) is defined as constant or intermittent lower, cyclical or non-cyclical abdominal pain of at least six months’ duration. In the United Kingdom, over 1 million women suffer from CPP, with an estimated annual healthcare cost above £150 million. The aetiology of CPP is unknown in up to 50% of women, and in the remainder, the symptoms of CPP is associated with endometriosis, pelvic adhesions, irritable bowel syndrome or painful bladder syndrome. CPP is often accompanied by painful periods, pain during sexual intercourse and defaecation. Fatigue, sleep disturbances and depression are also common among this group of women. CPP asserts a heavy emotional, social and economic burden. Standard treatments such as hormonal and analgesic regimens are often associated with unacceptable side effects, even if helpful for the pain, underlining an urgent need for a satisfactory treatment. The meridian balanced method (BM) electro-acupuncture (EA) treatment (acupuncture needling + traditional Chinese medicine health consultation [TCM HC]) may be effective in managing CPP symptoms. Thus, I have completed a pilot study comprising of a three-armed randomised controlled trial (RCT), using a mixed methods research (MMR) approach, to assess the feasibility of a future large-scale RCT to determine the effectiveness of the meridian BMEA treatment on CPP in women. My hypothesis is that it is feasible to conduct such a large-scale RCT for CPP in women. The primary objectives were to determine recruitment and retention rates. The secondary objectives were to evaluate the, acceptability of the methods of recruitment, randomisation, interventions and assessment tools and any signals of effectiveness of the interventions. Thirty (30) women with CPP were randomised into three groups: BMEA treatment, TCM HC, or National Health Service standard care (NHS SC) group. The effects of my interventions were assessed by validated pain, physical and emotional functioning questionnaires, completed at weeks 0, 4, 8 and 12 of the study. Semi-structured telephone interviews and focus group discussions to explore participants’ experience of the study were conducted. Of the 59 women who were referred to the study, 30 women (51%) were randomised. There was a statistically significant difference in retention rates between the three groups. The retention rates were 80% (95% CI 74-96), in the BMEA treatment group, 53 % (95% CI 36- 70) in the TCM HC group and 87% (95% CI 63-90) in the NHS SC group. (Chi-square test, p=0.08) The attendance rates of the BMEA treatment group were 90% compared to 56% in the TCM HC group. There was a statistically significant difference (Mann-Whitney test, p=0.023) in attendance between the two intervention groups. Telephone interviews regarding the acceptability of the methods of recruitment, randomisation, assessment tools and interventions were positive. No adverse effects that were directly related to BMEA treatments were reported or observed. A higher proportion of the BMEA treatment group achieved clinical significance in the VAS-pain, BPI-pain severity, interference, and sleep scores, when compared to the other two groups. Due to small sample sizes, there was insufficient power to show statistically significant difference. (Fishers Exact Test, p=1.0) Analyses of the questionnaire data per group showed statistically significant differences in the following: the BMEA treatment group experienced less in pain at weeks 4 (p=0.01) and 8 (p=0.005); less helplessness (p=0.03) and their anxiety and depression scores declined at week 4 (p=0.04). The NHS SC group also reported less pain at week 4 (p=0.04). However, this group scored higher in anxiety and depression at weeks 8 and 12 (p=0.04). No statistically significant differences were achieved between the three groups at baseline, weeks 4, 8 and 12 in all scores. The therapeutic benefits gained by the TCM HC group were less compared to those of the BMEA treatment group, but better when compared to the NHS SC group. The BMEA treatment and TCM HC groups showed lower scores in anxiety and depression while the NHS SC group showed higher scores in anxiety and depression. The NHS SC group also tended to ruminate and magnify their problems as well as feeling more helpless than the other two groups. The three key themes that emerged from thematic analysis of focus group discussions were the “whole person effects” where participants reported an improvement in pain, sleep and a general sense of wellbeing in the two intervention groups; the “experience of standard care” and “impact of living with CPP”. In conclusion, the results of my pilot study are supportive of the feasibility of a future large-scale study. There were signals of effectiveness of interventions but the sample size was too small to make a definitive conclusion.

L’interface hôte/microbiote intestinal est un système d’interactions complexes dont le déséquilibre est associé au développement des maladies inflammatoires chroniques. Les traitements anti-TNFα sont très efficaces dans ces maladies, mais seulement chez certains patients. L’objectif de ce travail était de rechercher un lien entre composition du microbiote intestinal et réponse aux traitements anti-TNFα dans deux types de maladies inflammatoires chroniques, les spondyloarthrites et les maladies inflammatoires chroniques de l’intestin. Nous avons retrouvé des variations de la composition du microbiote intestinal après traitement par anti-TNFα chez des patients atteints de spondyloarthrite et avons identifié un nœud taxonomique prédictif de la réponse thérapeutique à trois mois. Ce nœud taxonomique, l’ordre des Burkholderiales, étant ainsi un biomarqueur potentiellement utilisable en pratique clinique, nous avons déposé une demande de brevet européen, qui est en cours d’instruction. Ce travail a été poursuivi par un nouveau protocole de recherche clinique incluant des patients atteints de spondyloarthrites mais aussi de maladies inflammatoires chroniques intestinales. Ce protocole est financé par le CHU de Bordeaux dans le cadre de l’Appel d’Offre Interne. Il permettra de valider les hypothèses de notre premier travail, en réalisant notamment des PCR quantitatives utilisant des amorces spécifiques de l’ordre des Burkholderiales. Nous avons de plus retrouvé in vitro pour la première fois à notre connaissance une modulation de la croissance bactérienne chez Bacteroides fragilis en réponse au TNFα humain. / The host/gut microbiota interface is a system of complex interactions whose imbalance is associated with the development of chronic inflammatory diseases. Anti-TNFα treatments are very effective in these diseases, but only in some patients. The purpose of this work was to find a link between the composition of the intestinal microbiota and clinical response to anti-TNFα treatments in two types of chronic inflammatory diseases, spondyloarthritis and inflammatory bowel disease. We found variations in the composition of the intestinal microbiota after treatment with anti-TNFα in patients with spondyloarthritis and identified a taxonomic node predictive of the therapeutic response at 3 months. This taxonomic node, the Burkholderiales order, being a biomarker potentially usable in clinical practice, we have filed a European patent application, which is currently under investigation. This work was continued by a new clinical research protocol including patients with spondyloarthritis but also with inflammatory bowel diseases. This protocol is funded by the Bordeaux University Hospital as part of the internal call for tenders. It will validate the hypotheses of our first work, notably by performing quantitative PCRs using specific primers targeting the order of Burkholderiales. In vitro, we have also found for the first time, to our knowledge, a modulation of bacterial growth in Bacteroides fragilis in response to human TNFα.

A adesão ao tratamento medicamentoso é um importante fator determinante no sucesso terapêutico. A adesão do paciente pode ser influenciada por fatores diversos, ligados à doença, ao tratamento, ao paciente, às condições sociais e econômicas, como também, relacionada ao sistema de saúde que o atende. Ainda que existam inúmeros estudos sobre a adesão ao tratamento em portadores com doenças crônicas, são escassas as investigações sobre este tema nas doenças inflamatórias intestinais. O presente trabalho teve o objetivo de avaliar a adesão ao tratamento medicamentoso e os possíveis fatores que a influenciam, em pacientes portadores de doenças inflamatórias intestinais (DII): doença de Crohn (DC) e retocolite ulcerativa (RCU), do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP), da Universidade de São Paulo. Como controles, foram investigados pacientes portadores de pancreatite crônica e insuficiência pancreática (PC), com medicação fornecida pelo hospital, tal como os pacientes portadores de DII. Foram também investigados pacientes portadores de afecções digestivas variadas (ADV), grupo no qual a medicação prescrita não era fornecida pelo hospital. Por meio de estudo transversal e indireto, foi realizada entrevista estruturada para avaliar a adesão de 110 pacientes, que foram caracterizados como apresentando maior ou menor grau de adesão. Esta classificação foi baseada no cotejo entre os dados do prontuário e os informados pelo paciente em entrevista, considerando a afirmação do paciente que usava a medicação e que conhecia o nome da droga em uso. Utilizou-se, também, o teste de Morisky, que permite avaliar o padrão de comportamento do paciente em relação ao uso diário do medicamento. Este teste consiste de quatro perguntas padronizadas relacionadas ao esquecimento, descuido com o horário de tomada do medicamento, percepção de efeitos colaterais e ausência de sintomas. Na análise dos medicamentos utilizados pelo paciente, foram observadas as seguintes proporções de pacientes classificados como menos aderentes: 15,4% em pacientes portadores de DC, 13,3% na RCU, 8,4% na PC e 16,6% nos pacientes do grupo ADV. Porém, o teste de Morisky mostrou as seguintes proporções de menos aderentes: 50% de pacientes portadores de DC, 63,3% na RCU, 54,2% na PC e 63,4% na ADV. Não houve diferenças estatisticamente significativas, entre os grupos de pacientes, tanto na análise dos medicamentos utilizados pelo paciente como pelos resultados do teste de Morisky. Em análise univariada, nenhum dos fatores demográficos, sociais, clínicos ou referentes ao tratamento medicamentoso apresentou relação estatisticamente significativa, comum a todos os grupos, que indicasse influência sobre a adesão ao tratamento. Apesar do alto grau de adesão, de acordo com a análise dos medicamentos utilizados, detectou-se alto percentual de não-adesão ao tratamento medicamentoso ligado ao comportamento habitual e independente do diagnóstico, ou do acesso gratuito aos medicamentos. Isto pode indicar a existência de um padrão específico de comportamento dos usuários do serviço, o que sugere a necessidade de maior atenção dos profissionais de saúde para o problema, bem como medidas de educação do paciente quanto ao uso dos medicamentos. / Compliance to drug therapy is an important factor determining a successful treatment. Patient compliance may be influenced by various factors related to the disease, to treatment, to the patient himself, to his socioeconomic condition, as well as to the health system. Although many studies have assessed compliance to treatment in patients with chronic diseases, few investigations are available in inflammatory bowel diseases (IBD). The objective of the present study was to assess compliance to drug therapy in patients with IBD - Crohn’s disease (CD) and ulcerative colitis (UC), seen at the University Hospital, Faculty of Medicine of Ribeirão Preto (HCFMRP), University of São Paulo, Brazil. Patients with chronic pancreatitis (CP) and pancreatic insufficiency who received free medication supplied by the hospital, like the IBD patients, were used as controls. Patients with various digestive affections (VDA) whose prescribed medication was not supplied by the hospital were also investigated. In a transverse and indirect study, a structured interview was applied to assess the compliance of 110 patients, who were characterized as presenting a higher or lower degree of compliance. This classification was based on a comparison of data in the medical records to the information provided by the patient in the interview, considering the patient’s statements that he/she actually used the medication and was capable of produce correctly its name. The Morisky test was also used to assess the behavioral pattern of the patient regarding the daily use of the medication. This test consists of four standardized questions that evaluate forgetfulness, carelessness regarding the time when the medication should be taken, the perception of side effects, and the absence of symptoms. In the analysis of patient statements on medication in use, the proportions of patients regarded as less compliant were as follows: 15.4% of patients with CD, 13.3% of those with UC, 8.4% of those with CP, and 16.6% of those with VDA. However, the Morisky test revealed the following proportions of less compliant patients: 50% of patients with CD, 63.3% of those with UC, 54.2% of those with CP, and 63.4% of those with VDA. No statistically significant differences were observed between the four groups regarding evaluation according to either the analysis of patient statements or the results of the Morisky test. Univariate analysis revealed that none of the demographic, social, or clinical factors or the variables related to drug therapy showed statistically significant relationships, common to all groups, that would indicate their influence on compliance to treatment. Despite the high degree of compliance evaluated by patient statements on medication in use, a high degree of noncompliance to treatment linked to habitual behavior was detected. Those findings were independent on either disease type or free access to medication. This may indicate the existence of a specific behavioral pattern common to the local health system users, which suggests the need for better consideration of the problem on the part of the health professionals, as well as the need for measures of patient education regarding medication use.

INTRODUÇÃO: O transplante de intestino (TI) estabeleceu-se como tratamento para pacientes com falência intestinal e complicações da nutrição parenteral. Entretanto, sepse continua sendo a principal causa de mortalidade. A lesão causada pela isquemia seguida de reperfusão (LIR) é apontada como um dos fatores de ruptura da barreira mucosa intestinal, com consequente translocação bacteriana e sepse, seja precocemente, por lesão epitelial direta, seja mais tardiamente pela sua associação com o desenvolvimento da rejeição celular aguda. Criou-se um modelo de TI em porcos jovens com a finalidade de estudar a LIR e seus efeitos no epitélio intestinal. MÉTODOS: Para a padronização do modelo, foram realizados 25 procedimentos, tendo sido testados os tamanhos dos animais, as soluções de preservação, o tipo de drenagem venosa, o tipo de reconstrução intestinal e o tempo de duração do experimento. Na pesquisa propriamente dita, 20 porcos jovens foram submetidos a TI ortotópico. Dois grupos foram determinados conforme o tempo de isquemia fria a que foi submetido o intestino: grupo 1 (n=12) 90 minutos (min) e grupo 2 (n=8) 180 min. O procedimento foi realizado sob técnica asséptica e as anastomoses vasculares realizadas entre a aorta do doador e a aorta infra-renal do receptor e a veia porta do doador e a veia cava inferior do receptor. O trânsito intestinal foi reconstruído através de anastomoses entre o jejuno proximal do doador e do receptor e o íleo terminal do doador e do receptor. A solução de preservação utilizada foi Euro Collins. Não foi administrada medicação imunossupressora, exceto pela metilprednisolona (20mg/kg) no momento da reperfusão. Fragmentos de intestino foram obtidos: 1 no momento da laparotomia do doador, o fragmento basal, considerado controle, 2 30 min após a reperfusão e 3 3 dias após o transplante. Os fragmentos assim obtidos foram submetidos a: 1 análise histológica com coloração de hematoxilina-eosina (HE), 2 análise imunoistoquímica para a detecção de infiltração da mucosa por neutrófilos (marcados pelos grânulos ricos em mieloperoxidase MPO), 3 análise histoquímica para quantificação de células epiteliais em apoptose pelo método TUNEL, 4 análise da expressão dos genes da endotelina-1 (ET-1) e da interleucina-6 (IL-6), do gene antiapoptótico Bcl-XL e do gene pró-apoptótico Bak. A análise estatística foi realizada utilizando-se o teste de Mann-Whitney na comparação entre os grupos e, na análise da evolução temporal da LIR em cada grupo, o teste de Friedman seguido do teste post hoc de Dunn quando detectada diferença estatisticamente significante. RESULTADOS: Não foram encontradas diferenças entre os grupos quanto às alterações histológicas estudadas. O grau de infiltração da mucosa por neutrófilos elevou-se significantemente nos dois grupos 30 min após perfusão, tendo persistido elevado 3 dias após o TI somente no grupo 2. O número de células epiteliais em apoptose detectadas pelo método TUNEL sofreu incremento significante apenas no grupo 1, 3 dias após o procedimento. As duas citocinas estudadas, IL-6 e ET-1 mostraram elevação significante 30 minutos após a reperfusão, tendo retornado aos níveis basais 3 dias após a cirurgia em ambos os grupos. Detectou-se redução significante da expressão do Bcl-XL somente no grupo 1, 3 dias após o TI. CONCLUSÕES: As citocinas estudadas estão envolvidas no processo de LIR nas fases iniciais do TI. Ocorre diminuição da expressão de gene anti-apoptótico e aumento do número de células em processo de morte celular de maneira mais intensa no grupo submetido a menor tempo de isquemia / INTRODUCTION: Intestinal transplantation (ITx) has become an accepted mode of treatment of intestinal failure patients who develop parenteral nutrition-related complications. Overall outcomes have dramatically improved but sepsis remains the leading cause of mortality. Ischemia-reperfusion injury (IRI) has been related to the development of sepsis due either to direct mucosal damage or to increased risk of acute cellular rejection. An experimental ITx model has been idealized in order to better characterize IRI-associated mucosal damage. METHOD: 25 procedures involving 75 outbred pigs were necessary to standardize the procedure. Weight of the animals, venous drainage, intestinal transit reconstruction as well as the time period the animals should be maintained alive were evaluated. Orthotopic ITx was performed in 20 hybrid pigs. Two groups were assigned according to cold ischemia time (CI): group 1 (n=12) 90 minutes (min), group 2 (n=8) 180 min. The procedure was performed under aseptic technique and portal drainage was adopted as standard. Intestinal transit reconstruction involved the performance of termino-terminal anastomosis between donor and recipient jejunum and donor and recipient terminal ileum. Euro-Collins was used as preservation solution. 20mg/kg of metilprednisolone was administered at reperfusion and no other immunosuppressive drug has been employed. Specimens were collected from the donor at laparotomy, and from the receptor, 30 min, and 3 days after reperfusion. Mucosal damage was assigned by histological evaluation with hematoxylin-eosin dye. Neutrophilic infiltration was quantified using myeloperoxidase (MPO) immunohistochemical assay and epithelial cell apoptosis was also assigned by means of TUNEL assay. Molecular biology involved the quantification of the expression of the IL-6, ET-1, Bak, and Bcl-XL genes. RESULTS: No statistical difference was detected between the groups as far as plain histological evaluation is regarded. Neutrophilic infiltration increased in a similar fashion in both groups, but lasted longer in group 2. Apoptosis detected by TUNEL showed significant increase in group 1, 3 days after surgery. Anti-apoptotic gene Bcl-XL had its expression decreased in group 1, in 3 days as well. Endothelin-1 and IL-6 genes expression increased 30 min after the procedure and had already returned to baseline 3 d after surgery. CONCLUSION: IL-6 and ET-1 are involved precociously in the development of intestinal IRI. Neutrophilic infiltration lasted longer in the group submitted to longer CI. Although there were no significant differences between the groups, significant increase in the number of apoptotic epithelial cells 3 days after reperfusion could be detected in animals

Les maladies inflammatoires chroniques de l'intestin (MICI) sont caractérisées par une sécrétion exagérée de cytokines pro-inflammatoires, une hyperactivation des lymphocytes T effecteurs (Teff) et un contrôle insuffisant par les lymphocytes T régulateurs (Treg). Nous avons montré que le traitement par anti-TNFbetas'accompagne d'une augmentation significative des Treg Foxp3+ dans le sang de patients atteints de MICI en poussée. L'infliximab est associé à une potentialisation de leur fonction suppressive. Dans un 2ème travail nous montrons que les Treg sont incapables de prévenir la colite même s'il existe une augmentation du nombre de Treg dans les ganglions mésentériques ainsi qu'une majoration du nombre et de la fonction suppressive ex-vivo des Treg issus du colon inflammatoire. Cet impact fonctionnel positif sur les Treg issus des ganglions mésentériques était sélectif de la sous-population de Treg NRP1- correspondant aux Treg induits. La diminution significative aussi bien in vitro qu'in vivo de la conversion des LT naïfs en Treg en condition inflammatoire contribue probablement à leur incapacité à contenir la colite. Dans un 3ème travail, nous avons étudié la dynamique des LT Th1, Th17 et Treg et des sous-populations de LT CD4+ co-exprimants IL-17/IFNgamma, IL-17/Foxp3 et IFNgamma/Foxp3 chez des patients MICI en rémission clinique suivis tous les 3 mois. Une élévation des LT Foxp3/IL-17 du sang précédait la rechute de la maladie suggérant un rôle potentiel pathogénique de cette sous-population de LT. Ces éléments illustrent les concepts de conversion et plasticité des Treg au cours des MICI mais aussi leur rôle comme cible pour optimiser et développer de nouvelles biothérapies / Inflammatory bowel disease (IBD) are characterized by an excessive secretion of pro-inflammatory cytokines, hyperactivation of effector T cells (Teff) and insufficient control by regulatory T cells (Treg). We showed that treatment with anti-TNFbeta is accompanied by a significant increase in Foxp3+ Treg in the blood of patients with IBD. Infliximab is also associated with a potentiation of Treg suppressive function. In a second study, we showed that Treg are unable to completely prevent colitis, even as we have shown a significant increase in the number of Treg in the mesenteric lymph nodes and also an increase in number and the ex-vivo suppressive function of Treg cells from the inflammatory colon. The positive impact of intestinal inflammation on the suppressive function of Treg from the mesenteric lymph nodes was selective in the sub-population of Treg NRP1- majority representing iTreg. The significant decrease both in vitro and in vivo neo-conversion of LT to naïve Treg in inflammatory conditions, contributing to the inability of Treg to contain colitis. In a third study, we systematically studied the dynamics of LT Th1, Th17 and Treg as well as subpopulations of CD4+ T cells that co-express IL-17/IFNgamma, IL-17/Foxp3 and IFNgamma/Foxp3 from a cohort of IBD patients in clinical remission followed every 3 months. A rise in the blood of a Treg Foxp3+ mixed population producing IL-17 preceded the onset of a relapse of IBD suggesting a pathogenic potential of this subpopulation of LT. All these elements illustrate the concepts of conversion and plasticity of Treg in IBD but also the key role of Treg as a target to optimize and develop new biological therapies

The inflammasome complex is a multiprotein complex that may play a role in the pathogenesis of inflammatory bowel disease (IBD) by secreting the inflammatory cytokines interleukin (IL)-1β and IL-18, and inducing pyroptosis, as a response to signals through several inflammasome sensors. This study looked at the expression of several inflammasome components in the ileum and colon of patients suffering from IBD. The inflammasome sensors NLRP1, NLRP3, AIM2 and pyrin were upregulated in whole intestinal tissue of IBD patients, particularly in the colon. NLRP6 expression was increased in the colon of Crohn's disease patients, but not ulcerative colitis patients relative to colon of controls, and was reduced in the ileum of Crohn's disease patients compared to control ileum. Expression of caspase-1 and IL-1β, but not IL-18, were also increased in ileum and colon tissue from Crohn's patients. To identify the cell type where inflammasome expression was altered in Crohn’s disease, transcription of inflammasome subunits in intestinal tissue enriched for epithelial cells or lamina propria (LP) cells was analysed. These analyses indicated that LP cells have greater expression of the inflammasome sensors NLRP1, NLRP3, AIM2 and pyrin relative to epithelial cells, both during disease and in control tissue. Moreover, LP cells from Crohn’s patients have higher expression level of NLRP1, AIM2 and pyrin than LP cells from controls. In contrast the inflammasome sensor NLRP6 was more highly expressed by epithelial cells relative to LP cells in general, and NLRP6 expression in LP cells from IBD patients was lower than that observed in LP cells from controls. The observed differential expression of inflammasome components in controls versus IBD intestine and in different cellular fractions of intestinal tissue highlight the importance of understanding the role of the inflammasome in IBD and hints at the possibility of targeting the inflammasome pathway as a future treatment strategy.

Orientador: Sérgio Alberto Rupp de Paiva / Resumo: A Doença Inflamatória Intestinal (DII) apresenta curso variável. O tratamento adequado da fase aguda pode levar à remissão clínica da doença, cujo seguimento ocorre ambulatorialmente. Nesta condição, os pacientes mantêm hábito de vida normal e alguns ainda permanecem com inflamação. Esta situação de doença e estilo de vida pode interferir na composição corporal e no estado nutricional dos pacientes. Doença de Crohn (DC) e Retocolite Ulcerativa (RCU) são doenças heterogêneas em diversos aspectos, dentre eles na composição corporal e no estado nutricional. O objetivo do estudo foi avaliar clusters relacionados à variação do estado nutricional na DIl. Foi realizado estudo transversal com pacientes com DII, submetidos a avaliação clínica (Crohn’s Disease Activity Index (CDAI), escore de Mayo e uso de medicações), nutricional (recordatório de 24 horas, International Physical Activity Questionnaire (IPAQ), peso, estatura, Índice de Massa Corporal (IMC), absorciometria por raios-X de dupla energia (DEXA), análise de bioimpedância elétrica (BIA), força de preensão manual, teste de caminhada de 6 minutos) e laboratorial (hemoglobina (Hb), hematócrito (Ht), Proteína C Reativa (PCR), velocidade de hemossedimentação (VHS), albumina). Com base nestes parâmetros, foram diagnosticados com Desnutrição (Global Leadership Initiative on Malnutrition, GLIM) e Sarcopenia (European Working Group on Sacopenia in Older People 2, EWGSOP2). Foram realizados testes estatísticos descritivos por meio do ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Inflammatory Bowel Disease (IBD) presents a variable course. Adequate treatment of the acute phase may lead to clinical remission of the disease, which is followed in the outpatient clinic. In this condition, patients maintain normal life habit and some still remain with inflammation. Both disease and lifestyle situation may interfere with the body composition and nutritional status of the patients. Crohn's disease (CD) and ulcerative colitis (UC) are heterogeneous diseases in several aspects, including body composition and nutritional status. The aim of the study was to evaluate clusters related to the variation of the nutritional status in IBD. A cross-sectional study was performed with IBD patients, by clinical (Crohn's Disease Activity Index (CDAI), Mayo score and medications), nutritional (24-hour recall, International Physical Activity Questionnaire (IPAQ), weight, stature, Body Mass Index (BMI), dual energy X-ray absorptiometry (DEXA), electrical bioimpedance (BIA), handgrip force, 6-minute walk test) and laboratorial evaluation (hemoglobin (Hb), hematocrit (Ht), C-Reactive Protein (CRP), erythrocyte sedimentation rate (ESR), albumin). Based on these parameters, were diagnosed Malnutrition (Global Leadership Initiative on Malnutrition (GLIM) and Sarcopenia (European Working Group on Sacopenia in Older People 2, EWGSOP2). Descriptive statistical tests were performed by mean ± standard deviation for numerical variables with normal or median distribution and quartiles (Q1... (Complete abstract click electronic access below) / Doutor

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