190 research outputs found

    Germinal centers in human lymph nodes contain reactivated memory B cells

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    To reveal migration trails of antigen-responsive B cells in lymphoid tissue, we analyzed immunoglobulin (Ig)M-VH and IgG-VH transcripts of germinal center (GC) samples microdissected from three reactive human lymph nodes. Single B cell clones were found in multiple GCs, one clone even in as many as 19 GCs. In several GCs, IgM and IgG variants of the same clonal origin were identified. The offspring of individual hypermutated IgG memory clones were traced in multiple GCs, indicating repeated engagement of memory B cells in GC reactions. These findings imply that recurring somatic hypermutation progressively drives the Ig repertoire of memory B cells to higher affinities and infer that transforming genetic hits in non-Ig genes during lymphomagenesis do not have to arise during a single GC passage, but can be collected during successive recall responses

    Direct coupling of a free-flow isotachophoresis (FFITP) device with electrospray ionization mass spectrometry (ESI-MS)

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    We present the online coupling of a free-flow isotachophoresis (FFITP) device to an electrospray ionization mass spectrometer (ESI-MS) for continuous analysis without extensive sample preparation. Free-flow-electrophoresis techniques are used for continuous electrophoretic separations using an electric field applied perpendicular to the buffer and sample flow, with FFITP using a discontinuous electrolyte system to concurrently focus a target analyte and remove interferences. The online coupling of FFITP to ESI-MS decouples the separation and detection timeframe because the electrophoretic separation takes place perpendicular to the flow direction, which can be beneficial for monitoring (bio)chemical changes and/or extensive MSn studies. We demonstrated the coupling of FFITP with ESI-MS for simultaneous concentration of target analytes and sample clean-up. Furthermore, we show hydrodynamic control of the fluidic fraction injected into the MS, allowing for fluidically controlled scanning of the ITP window. Future applications of this approach are expected in monitoring biochemical changes and proteomics151734953502FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/06625-2; 2011/02477-3Alexander von Humboldt Foundation; Central European Institute of Technology (CEITEC

    Methodologies in Development Studies:An Overview

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    The interdisciplinary nature of the field of Development Studies (DS) makes it hard to point towards a ‘signature’ methodology. Different development challenges bring different ideas about what the problem is (ontology) and how researchers can know about it (epistemology), as well as different research methods. The rationale for choosing a method can be ideological or pragmatic. In the field of DS, this often entails knowing what methods research commissioners see as credible and what types of evidence they find persuasive. The weight placed on the data generated by certain methods and the lack of critical attention to how it was actually produced shows the importance of a focus on methodology. In looking at, or for, the defining methodologies of DS, this chapter focuses on methodology in a relatively narrow sense: what types of sample and what combinations of methods are typically used by researchers within DS to construct credible arguments around questions of policy or practice. It describes which methodologies constitute the bulk of DS research through analysis of projects and outputs. Finally, it asks what people who generate and use DS research could do to increase its rigour and relevance (Gujit and Roche, 2014; see also Oswald, Leach and Gaventa, this volume) and how the political economy of development research funding might militate against this

    Ventilator-associated pneumonia in children after cardiac surgery in The Netherlands

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    We conducted a retrospective cohort study in an academic tertiary care center to characterize ventilator-associated pneumonia (VAP) in pediatric patients after cardiac surgery in The Netherlands. All patients following cardiac surgery and mechanically ventilated for ≥24 h were included. The primary outcome was development of VAP. Secondary outcomes were duration of mechanical ventilation and length of ICU stay. A total of 125 patients were enrolled. Their mean age was 16.5 months. The rate of VAP was 17.1/1,000 mechanical ventilation days. Frequently found organisms were Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and Pseudomonas aeruginosa. Patients with VAP had longer duration of ventilation and longer ICU stay. Risk factors associated with the development of VAP were a PRISM III score of ≥10 and transfusion of fresh frozen plasma. The mean VAP rate in this population is higher than that reported in general pediatric ICU populations. Children with VAP had a prolonged need for mechanical ventilation and a longer ICU sta

    Initial microbial spectrum in severe secondary peritonitis and relevance for treatment

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    This study aims to determine whether abdominal microbial profiles in early severe secondary peritonitis are associated with ongoing infection or death. The study is performed within a randomized study comparing two surgical treatment strategies in patients with severe secondary peritonitis (n = 229). The microbial profiles of cultures retrieved from initial emergency laparotomy were tested with logistic regression analysis for association with ‘ongoing infection needing relaparotomy’ and in-hospital death. No microbial profile or the presence of yeast or Pseudomonas spp. was related to the risk of ongoing infection needing relaparotomy. Resistance to empiric therapy for gram positive cocci and coliforms was moderately associated with ongoing abdominal infection (OR 3.43 95%CI 0.95–12.38 and OR 7.61, 95%CI 0.75–76.94). Presence of only gram positive cocci, predominantly Enterococcus spp, was borderline independently associated with in-hospital death (OR 3.69, 95%CI 0.99–13.80). In secondary peritonitis microbial profiles do not predict ongoing abdominal infection after initial emergency laparotomy. However, the moderate association of ongoing infection with resistance to the empiric therapy compels to more attention for resistance when selecting empiric antibiotic coverage

    The benefits of participatory methodologies to develop effective community dialogue in the context of a microbicide trial feasibility study in Mwanza, Tanzania

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    BACKGROUND: As part of a microbicide trial feasibility study among women at high-risk of HIV and sexually transmitted infections in Mwanza City, northern Tanzania we used participatory research tools to facilitate open dialogue and partnership between researchers and study participants. METHODS: A mobile community-based sexual & reproductive health service was established in ten city wards. Wards were divided into seventy-eight geographical clusters and representatives at cluster and ward level elected in a process facilitated by the projects Community Liaison Officer. A city-level Community Advisory Committee (CAC) with representatives from each ward was established. Workshops and community meetings at ward and city-level were conducted to explore project-related concerns using tools adapted from participatory learning and action techniques such as listing, scoring, ranking, chapatti diagrams and pair-wise matrices. RESULTS: Key issues identified included beliefs that blood specimens were being sold for witchcraft purposes; worries about specula not being clean; inadequacy of transport allowances; and delays in reporting laboratory test results to participants. To date, the project has responded by inviting members of the CAC to visit the laboratory to observe how blood and genital specimens are prepared; demonstrated the use of the autoclave to community representatives; raised reimbursement levels; introduced HIV rapid testing in the clinic; and streamlined laboratory reporting procedures. CONCLUSIONS: Participatory techniques were instrumental in promoting meaningful dialogue between the research team, study participants and community representatives in Mwanza, allowing researchers and community representatives to gain a shared understanding of project-related priority areas for intervention

    Social learning in LEADER: Exogenous, endogenous and hybrid evaluation in rural development

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    This paper considers the relationship between the centralised exogenous, institutions and the embedded, endogenous institutions of rural governance in Europe through an examination the evaluation procedures of the European LEADER programme. LEADER is presented in the literature as progressive in terms of innovation and stakeholder engagement. Yet while the planning and management of LEADER embraces heterogeneity and participation, programmatic evaluation is centralised and held at arms length from delivery organisations. The paper reviews previous efforts to improve evaluation in LEADER and considers alternative strategies for evaluation, contrasting LEADER practice with participatory evaluation methodologies in the wider international context. Can evaluation in itself be valuable as a mode of social learning and hence a driver for endogenous development in rural communities in Europe? The paper concludes by examining the challenges in producing a hybrid form of evaluation which accommodates endogenous and exogenous values
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