Protocol for faecal microbiota transplantation in ulcerative colitis (FMTUC): a randomised feasibility study

Background The interaction of the gut microbiota with the human host is implicated in the pathogenesis of inflammatory and immunological diseases including ulcerative colitis (UC). Faecal microbiota transplantation (FMT) as a method of restoring gut microbial diversity is of increasing interest as a therapeutic approach in the management of UC. The current literature lacks consensus about the dose of FMT, route of administration and duration of response. Methods and analysis This single-blinded randomised trial will explore the feasibility of FMT in 30 treatment-naïve patients with histologically confirmed distal UC limited to the recto-sigmoid region (up to 40 cm from the anal verge). This study aims to estimate the magnitude of treatment response to FMT under controlled conditions. The intervention (FMT) will be administered by rectal retention enema. It will test the feasibility of randomising patients to: (i) single FMT dose, (ii) five daily FMT doses or (iii) control (no FMT dose). All groups will receive standard antibiotic gut decontamination and bowel preparation before FMT. Recruitment will take place over a 24-month period with a 12-week patient follow-up. Trial objectives include evaluation of the magnitude of treatment response to FMT, investigation of the clinical value of metabolic phenotyping for predicting the clinical response to FMT and testing the recruitment rate of donors and patients for a study in FMT. This feasibility trial will enable an estimate of number of patients needed, help determine optimal study conditions and inform the choice of endpoints for a future definitive phase III study. Ethics and dissemination The trial is approved by the regional ethics committee and is sponsored by Abertawe Bro Morgannwg University’s Health Board. Written informed consent from all patients will be obtained. Serious adverse events will be reported to the sponsor. Trial results will be disseminated via peer review publication and shared with trial participants. Trial registration number ISRCTN58082603; Pre-results

In-Cell Biochemistry Using NMR Spectroscopy

Biochemistry and structural biology are undergoing a dramatic revolution. Until now, mostly in vitro techniques have been used to study subtle and complex biological processes under conditions usually remote from those existing in the cell. We developed a novel in-cell methodology to post-translationally modify interactor proteins and identify the amino acids that comprise the interaction surface of a target protein when bound to the post-translationally modified interactors. Modifying the interactor proteins causes structural changes that manifest themselves on the interacting surface of the target protein and these changes are monitored using in-cell NMR. We show how Ubiquitin interacts with phosphorylated and non-phosphorylated components of the receptor tyrosine kinase (RTK) endocytic sorting machinery: STAM2 (Signal-transducing adaptor molecule), Hrs (Hepatocyte growth factor regulated substrate) and the STAM2-Hrs heterodimer. Ubiquitin binding mediates the processivity of a large network of interactions required for proper functioning of the RTK sorting machinery. The results are consistent with a weakening of the network of interactions when the interactor proteins are phosphorylated. The methodology can be applied to any stable target molecule and may be extended to include other post-translational modifications such as ubiquitination or sumoylation, thus providing a long-awaited leap to high resolution in cell biochemistry

Regulation of ErbB2 Receptor Status by the Proteasomal DUB POH1

Understanding the factors, which control ErbB2 and EGF receptor (EGFR) status in cells is likely to inform future therapeutic approaches directed at these potent oncogenes. ErbB2 is resistant to stimulus-induced degradation and high levels of over-expression can inhibit EGF receptor down-regulation. We now show that for HeLa cells expressing similar numbers of EGFR and ErbB2, EGFR down-regulation is efficient and insensitive to reduction of ErbB2 levels. Deubiquitinating enzymes (DUBs) may extend protein half-lives by rescuing ubiquitinated substrates from proteasomal degradation or from ubiquitin-dependent lysosomal sorting. Using a siRNA library directed at the full complement of human DUBs, we identified POH1 (also known as Rpn11 or PSMD14), a component of the proteasome lid, as a critical DUB controlling the apparent ErbB2 levels. Moreover, the effects on ErbB2 levels can be reproduced by administration of proteasomal inhibitors such as epoxomicin used at maximally tolerated doses. However, the extent of this apparent loss and specificity for ErbB2 versus EGFR could not be accounted for by changes in transcription or degradation rate. Further investigation revealed that cell surface ErbB2 levels are only mildly affected by POH1 knock-down and that the apparent loss can at least partially be explained by the accumulation of higher molecular weight ubiquitinated forms of ErbB2 that are detectable with an extracellular but not intracellular domain directed antibody. We propose that POH1 may deubiquitinate ErbB2 and that this activity is not necessarily coupled to proteasomal degradation

Avaliação de um modelo de predição para apneia do sono em pacientes submetidos a polissonografia Evaluation of a prediction model for sleep apnea in patients submitted to polysomnography

OBJETIVO: Testar um modelo de predição para apneia do sono a partir de variáveis sociodemográficas e clínicas em uma população com suspeita de distúrbio do sono e submetida à polissonografia. MÉTODOS: Foram incluídos no estudo 323 pacientes consecutivos submetidos à polissonografia por suspeita clínica de distúrbio do sono. Utilizou-se um questionário com questões sociodemográficas e a escala de sonolência de Epworth. Foram medidos pressão arterial, peso, altura e SpO2. A regressão linear múltipla, tendo o índice de apneia-hipopneia (IAH) como variável dependente, foi utilizada para construir um modelo de predição de apneia do sono. A regressão logística multinomial foi realizada para verificar fatores associados de forma independente à gravidade da apneia (leve, moderada ou grave) em comparação à ausência de apneia. RESULTADOS: A prevalência de apneia do sono na população de estudo foi de 71,2%, e foi mais prevalente nos homens que nas mulheres (81,2% vs. 56,8%; p < 0,001). O modelo de regressão linear múltipla, com o log IAH como variável dependente, foi composto pelas seguintes variáveis independentes: circunferência do pescoço, apneia testemunhada, idade, IMC e presença de rinite alérgica. O melhor modelo de regressão linear encontrado conseguiu explicar 39% da variabilidade do IAH. Na regressão logística multinomial, a apneia leve esteve associada com IMC e circunferência do pescoço, e a apneia grave associou-se com idade, IMC, circunferência do pescoço e apneia testemunhada. CONCLUSÕES: Modelos de predição clínica para apneia do sono não substituem a polissonografia como ferramenta para o seu diagnóstico, mas podem otimizar sua indicação e aumentar a chance de positividade do exame.<br>OBJECTIVE: To test a prediction model for sleep apnea based on clinical and sociodemographic variables in a population suspected of having sleep disorders and submitted to polysomnography. METHODS: We included 323 consecutive patients submitted to polysomnography because of the clinical suspicion of having sleep disorders. We used a questionnaire with sociodemographic questions and the Epworth sleepiness scale. Blood pressure, weight, height, and SpO2 were measured. Multiple linear regression was used in order to create a prediction model for sleep apnea, the apnea-hypopnea index (AHI) being the dependent variable. Multinomial logistic regression was used in order to identify factors independently associated with the severity of apnea (mild, moderate, or severe) in comparison with the absence of apnea. RESULTS: The prevalence of sleep apnea in the study population was 71.2%. Sleep apnea was more prevalent in men than in women (81.2% vs. 56.8%; p < 0.001). The multiple linear regression model, using log AHI as the dependent variable, was composed of the following independent variables: neck circumference, witnessed apnea, age, BMI, and allergic rhinitis. The best-fit linear regression model explained 39% of the AHI variation. In the multinomial logistic regression, mild apnea was associated with BMI and neck circumference, whereas severe apnea was associated with age, BMI, neck circumference, and witnessed apnea. CONCLUSIONS: Although the use of clinical prediction models for sleep apnea does not replace polysomnography as a tool for its diagnosis, they can optimize the process of deciding when polysomnography is indicated and increase the chance of obtaining positive polysomnography findings