Targeting the non-structural protein 1 from dengue virus to a dendritic cell population confers protective immunity to lethal virus challenge

Dengue is the most prevalent arboviral infection, affecting millions of people every year. Attempts to control such infection are being made, and the development of a vaccine is a World Health Organization priority. Among the proteins being tested as vaccine candidates in preclinical settings is the non-structural protein 1 (NS1). In the present study, we tested the immune responses generated by targeting the NS1 protein to two different dendritic cell populations. Dendritic cells (DCs) are important antigen presenting cells, and targeting proteins to maturing DCs has proved to be an efficient means of immunization. Antigen targeting is accomplished by the use of a monoclonal antibody (mAb) directed against a DC cell surface receptor fused to the protein of interest. We used two mAbs (αDEC205 and αDCIR2) to target two distinct DC populations, expressing either DEC205 or DCIR2 endocytic receptors, respectively, in mice. The fusion mAbs were successfully produced, bound to their respective receptors, and were used to immunize BALB/c mice in the presence of polyriboinosinic: polyribocytidylic acid (poly (I:C)), as a DC maturation stimulus. We observed induction of strong anti-NS1 antibody responses and similar antigen binding affinity irrespectively of the DC population targeted. Nevertheless, the IgG1/IgG2a ratios were different between mouse groups immunized with αDEC-NS1 and αDCIR2-NS1 mAbs. When we tested the induction of cellular immune responses, the number of IFN-γ producing cells was higher in αDEC-NS1 immunized animals. In addition, mice immunized with the αDEC-NS1 mAb were significantly protected from a lethal intracranial challenge with the DENV2 NGC strain when compared to mice immunized with αDCIR2-NS1 mAb. Protection was partially mediated by CD4(+) and CD8(+) T cells as depletion of these populations reduced both survival and morbidity signs. We conclude that targeting the NS1 protein to the DEC205(+) DC population with poly (I:C) opens perspectives for dengue vaccine development.Brazilian National Research Council (CNPq)National Institute of Science and Technology Research Funding Agency (INCTV) - 15203*12São Paulo State Research Funding Agency (FAPESP) - 2007/08648-9, 2011/51761-6BNP-Paribas BankRio de Janeiro State Research Funding Agency (FAPERJ

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Analysis of the adaptive immune responses when a dengue virus antigen is targeted to two distinct populations of dendritic cells.

Células dendríticas são importantes na interação dos sistemas imune inato e adaptativo, apresentando antígenos para linfócitos T CD4+ e CD8+ e induzindo respostas adaptativas contra estes. Elas são alvo de interesse no desenvolvimento de novas estratégias vacinais que visam o direcionamento de antígenos para as diferentes populações de DCs in vivo. Consiste na administração de anticorpos contra receptores da superfície da DC em fusão com o antígeno. Nós comparamos o direcionamento da proteína NS1 de DENV2 para as duas principais populações de DCs no baço, na presença de diferentes agonistas. Poly I:C é capaz de induzir as respostas de Linfócitos T CD8+ mais robustas, mas somente quando o antígeno é direcionado para a população de DCs CD8+DEC205+. Com CpG observamos resposta de células T CD8+ equivalente quando a proteína foi direcionada para qualquer uma das populações estudadas. Direcionamento com Poly I:C gerou proteção contra desafio por DENV2 quando NS1 é direcionada para a população de DCs DEC205+, sendo esta proteção dependente de Linfócitos T CD8+ e CD4+.Dendritic Cells are critical in the interaction between innate and the adaptive immune system, presenting antigens to T lymphocytes and inducing adaptive immune responses against these. DCs are target for development of new vaccine strategies based on targeting antigens to different DCs populations in vivo. This consists on the administration of antibodies specific to DC surface endocytic receptor fused to an antigen. In this study, we evaluated the differences in adaptive responses when the NS1 protein from the DENV2 was directed to the two main populations of DCs in the spleen along with different agonistic molecules. Highest CD8+ T cell responses were seen when Poly I:C was used, but only when directed to the CD8+DEC205+ DC population. In the presence of agonist CpG we observed similar responses when the protein was targeted to either CD8+DEC205+ or CD8-DCIR2+ DCs. In the presence of Poly I:C, only targeting to the DEC205+ DCs was able to protect mice against a challenge with DENV2. This protection was dependent on both CD8+ and CD4+ T cell responses

Legitimacy and the authority of Law from the sociological perspective of Pierre Bourdieu

No Estado moderno se, por um lado, parece intrínseca a uma ordem jurídica a sua pretensão coercitiva em relação aos diferentes aspectos da vida social, resta evidente que tal ordem deve ser dotada de um atributo de autoridade que lhe garanta primazia em relação a outras ordens (morais, religiosas, pessoais), para que se justifique dotá-la de atributos tão singulares como a coercitividade. A essa autoridade, contudo, no contexto democrático - mais especificamente no contexto do Estado Democrático de Direito contemporâneo - cumpre dotar-se de uma legitimidade ou, ao menos, uma aparência de legitimidade suficiente para imprimir efetividade àquela autoridade. Nesse sentido, o presente trabalho busca redefinir, a partir da perspectiva bourdieuniana, a percepção dos principais argumentos de legitimidade e autoridade do Direito - visto não só enquanto ordenamento jurídico, mas principalmente a partir de sua produção no ambiente legislativo, tantas vezes ignorada pela doutrina jurídica que habitualmente toma o direito posto, já legislado, como ponto de partida, neutralizando as disputas do subcampo legislativo e ignorando, em ampla medida, a questão da legitimidade da produção normativa. A pesquisa demonstra, a partir da perspectiva da Teoria da Reprodução Social de Bourdieu & Passeron (1992) que coloca em cheque toda legitimidade, que é aqui vista como mero efeito de uma autoridade, como a própria democracia é um conceito em disputa, e que a legitimidade das ações legislativas, bem como a dos próprios agentes legislativos não pode ser pressuposta.If, in modern State, the coercive pretension seems intrinsic to the legal order in respect to all the different aspects of social life, it is also evident that such an order must have an authority which guarantees its primacy over other (such as moral, religious or personal orders), so that we are able to justify its singular attributes. To this authority, however, in the democratic context of contemporary State (under the Rule of Law), we must also have at least a notion of legitimacy, enough to give effectiveness to that authority. In this context, the present work seeks to redefine, from the perspective of Bourdieu\'s theory, the perception of the main arguments to law\'s legitimacy and authority - especially in aspects of the legislative work, usually overseen by juridical doctrine that takes law for granted, neutralizing the disputes in legislative field and ignoring to some extent the different critics to legislative legitimacy. Our research demonstrates, from the perspective of the Social Reproduction Theory, developed by Bourdieu & Passeron (1992) and which questions all legitimacy, seen as mere effect of authority, how democracy itself is a concept in dispute, and that the legitimacy of the legislative actions, and that of its agents, can\'t be taken for granted

Legitimacy and the authority of Law from the sociological perspective of Pierre Bourdieu

No Estado moderno se, por um lado, parece intrínseca a uma ordem jurídica a sua pretensão coercitiva em relação aos diferentes aspectos da vida social, resta evidente que tal ordem deve ser dotada de um atributo de autoridade que lhe garanta primazia em relação a outras ordens (morais, religiosas, pessoais), para que se justifique dotá-la de atributos tão singulares como a coercitividade. A essa autoridade, contudo, no contexto democrático - mais especificamente no contexto do Estado Democrático de Direito contemporâneo - cumpre dotar-se de uma legitimidade ou, ao menos, uma aparência de legitimidade suficiente para imprimir efetividade àquela autoridade. Nesse sentido, o presente trabalho busca redefinir, a partir da perspectiva bourdieuniana, a percepção dos principais argumentos de legitimidade e autoridade do Direito - visto não só enquanto ordenamento jurídico, mas principalmente a partir de sua produção no ambiente legislativo, tantas vezes ignorada pela doutrina jurídica que habitualmente toma o direito posto, já legislado, como ponto de partida, neutralizando as disputas do subcampo legislativo e ignorando, em ampla medida, a questão da legitimidade da produção normativa. A pesquisa demonstra, a partir da perspectiva da Teoria da Reprodução Social de Bourdieu & Passeron (1992) que coloca em cheque toda legitimidade, que é aqui vista como mero efeito de uma autoridade, como a própria democracia é um conceito em disputa, e que a legitimidade das ações legislativas, bem como a dos próprios agentes legislativos não pode ser pressuposta.If, in modern State, the coercive pretension seems intrinsic to the legal order in respect to all the different aspects of social life, it is also evident that such an order must have an authority which guarantees its primacy over other (such as moral, religious or personal orders), so that we are able to justify its singular attributes. To this authority, however, in the democratic context of contemporary State (under the Rule of Law), we must also have at least a notion of legitimacy, enough to give effectiveness to that authority. In this context, the present work seeks to redefine, from the perspective of Bourdieu\'s theory, the perception of the main arguments to law\'s legitimacy and authority - especially in aspects of the legislative work, usually overseen by juridical doctrine that takes law for granted, neutralizing the disputes in legislative field and ignoring to some extent the different critics to legislative legitimacy. Our research demonstrates, from the perspective of the Social Reproduction Theory, developed by Bourdieu & Passeron (1992) and which questions all legitimacy, seen as mere effect of authority, how democracy itself is a concept in dispute, and that the legitimacy of the legislative actions, and that of its agents, can\'t be taken for granted

Oceania Lurking: The Extremist “Menace” And Cultural Control

A “possível” emergência de partidos e políticos classificáveis como de extrema-direita ou extrema-esquerda é frequentemente compreendida como um movimento de avanço ou de retrocesso dos ideais democráticos ocidentais. Entretanto, as reações de medo ou esperança que tais movimentos provocam nos cidadãos e nos estudiosos – e a ampla cobertura midiática que têm – contrastam com outro avanço, esse inequívoco, da globalização econômica promovida pela ideologia neoliberal e do espraiamento do chamado “Estado Poiético”. Em sociedades onde o campo do poder está embebido da cultura/ideologia neoliberal, de propensões totalitárias, a espetacularização de tímidos avanços de partidos e políticos possivelmente contrários à ideologia dominante pode ser compreendida enquanto um mecanismo de cooptação e controle desses discursos possivelmente dissidentes ou subversivos à reprodução da cultura dominante. Por meio de uma política de medo e insegurança –contra pensamentos potencialmente dissidentes– relegam as sociedades à despolitização e à suposta técnica eficaz. Esse modelo estatal parece ser capaz de exercer um eficaz controle cultural, impedindo o exercício pleno da política e o florescimento de expressões culturais capazes de subverter seus pressupostos.The “possible” emergence of parties and politicians classifiable as extreme rightwing or extreme left-wing is frequently comprehended either as an advance to or a regress from western democratic ideals. However, fearful or hopeful reactions caused in citizens and scholars by such movements –and the wide media coverage they get– contrast with another advance, this one unequivocal, of economic globalization as promoted by neoliberal ideology and the spreading of the so called “Poietic State”. In societies where the field of power is impregnated with neoliberal culture/ideology, with its totalitarian propensities, the spectacularization of the timid advances of parties and politicians possibly contrary to dom-inant ideology might be comprehended as a mechanism of control and cooptation of such discourses possibly dissident or subversive to the reproduction of the dominant culture. Through a policy of fear and insecurity –from potentially dissident thoughts– they relegate those societies to depoliticization and supposedly efficient technique. This State model seems to be capable of exerting an effective cultural control, preventing the full exercise of politics and the blossoming of cultural expressions capable of subverting its core ideas

Evaluation of the diagnostic value of the tourniquet test in predicting severe dengue cases in a population from Belo Horizonte, State of Minas Gerais, Brazil

Introduction Dengue is prevalent in many tropical and sub-tropical regions. The clinical diagnosis of dengue is still complex, and not much data are available. This work aimed at assessing the diagnostic accuracy of the tourniquet test in patients with suspected dengue infection and its positivity in different classifications of this disease as reported to the Information System for Notifiable Disease in Belo Horizonte, State of Minas Gerais, Brazil between 2001 and 2006. Methods Cross-section analysis of the diagnostic accuracy of the tourniquet test for dengue, using IgM-anti-DENV ELISA as a gold standard. Results We selected 9,836 suspected cases, of which 41.1% were confirmed to be dengue. Classic dengue was present in 95.8%, dengue with complications in 2.5% and dengue hemorrhagic fever in 1.7%. The tourniquet test was positive in 16.9% of classic dengue cases, 61.7% of dengue cases with complications and 82.9% of cases of dengue hemorrhagic fever. The sensitivity and specificity of the tourniquet test were 19.1% and 86.4%, respectively. Conclusions A positive tourniquet test can be a valuable tool to support diagnosis of dengue where laboratory tests are not available. However, the absence of a positive test should not be read as the absence of infection. In addition, the tourniquet test was demonstrated to be an indicator of dengue severity