Search

HIV-1 envelope proteins (Envs) play a critical role in HIV infection. In a correct trimeric conformation, Env mediates virus–cell binding and fusion. Malfunctioning of this machinery renders virions incapable of infecting cells. Each HIV-1 virion carries 10–14 Envs, and therefore a defective Env may not necessarily render a HIV virion non-infectious, since other Env on the same virion may still be functional. Alternatively, it is possible that on a given virion either all the spikes are defective or all are functional. Here, we investigate Env conformations on individual virions using our new nanotechnology, “flow virometry”, and a panel of antibodies that discriminate between various Env conformations. We found that the majority of HIV-1 virions carry either only trimeric (“functional”) or only defective spikes. The relatively small subfraction of virions that carry both functional and nonfunctional Envs contributes little to HIV infection of human lymphoid tissue ex vivo. The observation that the majority of virions exclusively express either functional or nonfunctional forms of Env has important implications for understanding the role of neutralizing and non-neutralizing antibodies in the immune control of HIV infection as well as for the development of effective prophylactic strategies.

Background We compared safety and immunogenicity of intradermal (ID) vaccination with and without electroporation (EP) in a phase I randomized placebo-controlled trial of an HIV-DNA prime HIV-MVA boost vaccine in healthy Swedish volunteers. Methods HIV-DNA plasmids encoding HIV-1 genes gp160 subtypes A, B and C; Rev B; Gag A and B and RTmut B were given ID at weeks 0, 6 and 12 in a dose of 0.6 mg. Twenty-five volunteers received vaccine using a needle-free device (ZetaJet) with (n=16) or without (n=9) ID EP (Dermavax). Five volunteers were placebo recipients. Boosting with recombinant MVA-CMDR expressing HIV-1 Env, Gag, Pol of CRF01_AE (HIV-MVA) or placebo was performed at weeks 24 and 40. Nine of the vaccinees received a subtype C CN54 gp140 protein boost together with HIV-MVA. Results The ID/EP delivery was very well tolerated. After three HIV-DNA immunizations, no statistically significant difference was seen in the IFN-γ ELISpot response rate to Gag between HIV-DNA ID/EP recipients (5/15, 33%) and HIV-DNA ID recipients (1/7, 14%, p=0.6158). The first HIV-MVA or HIV-MVA+gp140 vaccination increased the IFN-γ ELISpot response rate to 18/19 (95%). CD4+ and/or CD8+ T cell responses to Gag or Env were demonstrable in 94% of vaccinees. A balanced CD4+ and CD8+ T cell response was noted, with 78% and 71% responders, respectively. IFN-γ and IL-2 dominated the CD4+ T cell response to Gag and Env. The CD8+ response to Gag was broader with expression of IFN-γ, IL-2, MIP-1β and/or CD107. No differences were seen between DNA vaccine groups. Binding antibodies were induced after the second HIV-MVA+/-gp140 in 93% of vaccinees to subtype C Env, with the highest titers among EP/gp140 recipients. Conclusion Intradermal electroporation of HIV-DNA was well tolerated. Strong cell- and antibody-mediated immune responses were elicited by the HIV-DNA prime and HIV-MVA boosting regimen, with or without intradermal electroporation use. Trial Registration International Standard Randomised Controlled Trial Number (ISRCTN) 60284968

As vantagens da utilização de jogos com objetivos educacionais podem ser consideradas como um consenso entre professores e alunos. No entanto, jogos sérios constituem-se objetos multimídia complexos e caros de se produzir. A natureza multidisciplinar dos jogos educacionais pressupõe o envolvimento e coordenação de uma equipe especializada. Com o objetivo de auxiliar no desenvolvimento de jogos educacionais é proposto o modelo computacional BROAD-PLG. O modelo é composto por uma arquitetura de alto nível, modelagens de domínio baseadas em características que descrevem três diferentes tipos dos jogos educacionais, e uma ferramenta de engenharia de aplicação, que permite instanciação de um arcabouço pronto para ser utilizado no desenvolvimento desses tipos de jogos. A separação de interesses divide o domínio de jogos educacionais em conjuntos de características que englobam aspectos educacionais, mecânica de jogos e elementos de gamificação. Ao final do trabalho são construídos três exemplos distintos de aplicações demonstrando esses três aspectos. Avaliando-se os exemplos pode-se concluir que o BROAD-PLG, apesar de estar na versão inicial, demonstra grande potencial para ser utilizado como uma ferramenta tanto na forma de geração de aplicações, quanto como referência na modelagem de domínio do problema. The advantages of using games for educational purposes can be considered as a consensus among teachers and students. However, serious games constitute complex and expensive to produce multimedia objects. The multidisciplinary nature of educational games requires the involvement and coordination of a specialized team. With the objective of assisting in the development of educational games is proposed the computational model BROAD-PLG. The model consists of a high-level architecture, domain modeling based on features that describe three different faces of educational games and application engineering tool that allows instantiation of a framework ready to be used in game development. The separation of concerns splits the domain of educational games into sets of features that include educational aspects of game mechanics and gamification elements. At the end of the work three different application examples are constructed demonstrating these three aspects. In reviewing the examples it can be concluded that the broad-PLG despites being in the initial stage shows great potential to be used as a tool in generating applications as reference in modeling the problem domain.

During Drosophila oogenesis, activation of Notch signaling in the follicular epithelium (FE) around stage 6 of oogenesis is essential for entry into the endocycle and a series of other changes such as cell differentiation and migration of subsets of the follicle cells. Notch induces the expression of zinc finger protein Hindsight and suppresses homeodomain protein Cut to regulate the mitotic/endocycle (ME) switch. Here we report that broad (br), encoding a small group of zinc-finger transcription factors resulting from alternative splicing, is a transcriptional target of Notch nuclear effector Suppressor of Hairless (Su(H)). The early pattern of Br in the FE, uniformly expressed except in the polar cells, is established by Notch signaling around stage 6, through the binding of Su(H) to the br early enhancer (brE) region. Mutation of the Su(H) binding site leads to a significant reduction of brE reporter expression in follicle cells undergoing the endocycle. Chromatin immunoprecipitation results further confirm Su(H) binding to the br early enhancer. Consistent with its expression in follicle cells during midoogenesis, loss of br function results in a delayed entry into the endocycle. Our findings suggest an important role of br in the timing of follicle cell development, and its transcriptional regulation by the Notch pathway.

Aquest article és el primer d'una sèrie de tres articles sobre qualitat en traducció. Els articles se succeeixen entre si i tracten la definició de traducció, qualitat i finalment qualitat en traducció. En relació amb el significat de traducció, en la literatura dels estudis de traducció s'han proposat moltes definicions. No obstant això, aquest article no és una revisió de la bibliografia. Per contra, suposa una contribució que consisteix a descriure un "paisatge" en què diverses definicions de traducció es poden "sembrar". Aquest paisatge està delimitat per dos eixos, que són l'abast i les especificacions, i proporciona un marc per discutir qüestions com a) la relació entre traducció i localització i b) el paper de les especificacions en traducció. Dins d'aquest paisatge, aquest article presenta dues postures oposada sobre la definició de traducció. Els autors no arriben a un acord sobre quina d'aquestes postures és més útil, però coincideixen que les parts interessades han de definir amb claredat el concepte de traducció abans de poder tractar la noció de qualitat en traducció de forma significativa. This article is the first in a three-part series on translation quality. The articles build on one another by addressing definitions of translation, quality, and finally translation quality. Concerning the meaning of translation, many definitions have been proposed in the translation-studies literature. This article, however, is not a literature review. Instead, it describes a "landscape" within which various definitions of translation can be "planted." This landscape is delimited by two axes-namely, scope and specifications-and provides a framework for discussing such questions as (a) the relationship between translation and localization and (b) the role of translation specifications. Within that landscape, this article presents two opposing views about the definition of translation. The authors disagree about which of these views is most useful, but they agree that stakeholders must clearly define translation before they can meaningfully address translation quality. Este artículo es el primero de una serie de tres artículos sobre calidad en traducción. Losartículos se suceden entre sí y tratan la definición de traducción, calidad y finalmente calidaden traducción. En relación con el significado de traducción, en la literatura de los estudiosde traducción se han propuesto muchas definiciones. No obstante, este artículo noes una revisión de la bibliografía. Por el contrario, supone una contribución que consisteen describir un "paisaje" en el que varias definiciones de traducción se pueden "sembrar".Este paisaje está delimitado por dos ejes, que son alcance y especificaciones, y proporcionaun marco para discutir cuestiones como a) la relación entre traducción y localizacióny b) el papel de las especificaciones en traducción. Dentro de este paisaje, este artículopresenta dos posturas opuesta sobre la definición de traducción. Los autores no llegan aun acuerdo sobre cuál de estas posturas es más útil, pero coinciden en que las partes interesadasdeben definir con claridad el concepto de traducción antes de poder tratar lanoción de calidad en traducción de forma significativa.

Peptide mimics of epitopes for pathogen-specific antibodies present in patient sera can be selected based on the phage display technology. Such mimotopes potentially represent vaccine candidates in case they are able to induce neutralizing antibodies upon vaccination. Here we analyze the immunogenicity of different conjugates of epitope EC26-2A4 localizing to the membrane proximal external region (MPER) of the HIV-1 transmembrane protein gp41. The EC26-2A4 epitope, which overlaps with that of the broadly neutralizing monoclonal antibody (mAb) 2F5, was coupled to sequential oligopeptide carriers (SOC) or to palmitoyl acid for better immunogenicity. Upon prime-boost immunizations of mice, the peptide conjugates induced EC26-2A4 specific antibodies in all settings and mice sera neutralized HIV-1SF162.LS in standardized neutralization assays. Thus, the EC26-2A4 MPER epitope represents a promising vaccine candidate for further analysis in larger animals with respect to the breadth of the neutralizing antibodies induced. (C) 2012 Elsevier Ltd. All rights reserved. Vaccine

International audience; Cyclic-beta-glucans (C beta G) consist of cyclic homo-polymers of glucose that are present in the periplasmic space of many Gram-negative bacteria. A number of studies have demonstrated their importance for bacterial infection of plant and animal cells. In this study, a mutant of Rhizobium ( Sinorhizobium) sp. strain NGR234 (NGR234) was generated in the cyclic glucan synthase ( ndvB )-encoding gene. The great majority of C beta G produced by wild-type NGR234 are negatively charged and substituted. The ndvB mutation abolished C beta G biosynthesis. We found that, in NGR234, a functional ndvB gene is essential for hypo-osmotic adaptation and swimming, attachment to the roots, and efficient infection of Vigna unguiculata and Leucaena leucocephala.

The theoretical approaches to parliamentary coalitions that exist in international political science are presented in the article. A distinction between different types of coalitions is made, coalitions are analyzed in the parliaments of Poland and Ukraine after the parliamentary elections of 2005 and 2006 respectively. The attention is paid to the difficulties in determining the type of coalition in Ukraine due to lack of clear rules of the political game in Ukraine.

La investigación que se presenta en este artículo persigue construir conocimiento sobre cómo se percibe, se trata, se transforma y evoluciona el capital cultural adquirido por los asesores en los Centros de Profesorado una vez que éstos regresan a sus destinos en los centros de primaria y secundaria. Nuestro trabajo se construyó sobre el extenso relato de doce ex – asesores de CEP acerca de sus concepciones, creencias y visiones sobre cinco grandes temas: educación/enseñanza, escuela como organización, Administración educativa, relaciones con los colegas en el centro, y las relaciones con los alumnos y sus familias. Se trataba de constatar si, en efecto, podíamos hablar o no de un capital cultural adquirido en el CEP, y en qué medida esa acumulación de aprendizaje y experiencia había hecho evolucionar e incluso cambiar de modo sustancial las posiciones y concepciones de nuestros sujetos sobre la enseñanza y lo escolar., The research project that is partially presented in this paper sought to increase our knowledge about the ways in which the cultural capital gained by agents of external support to schools in Teachers’ Centers (CEP) is perceived, dealt with and transformed once they go back to their teaching duties in primary and secondary schools. Field work consisted of in-depth interviews to twelve ex – agents of external support, covering their auto-biographical experience, beliefs, views and conceptions on five broad issues: education/teaching, school as an organization, educational administration, relationship with fellow teachers in schools, and relationships with students and parents. The aim was to ascertain if it was in fact warranted to talk about a cultural capital acquired in Teachers’ Centers (CEP), and to what extent such stock of learning and experience had shaped or even radically change the views and beliefs held by the participating subjects., Grupo de Investigación FORCE Universidad de Granada

International audience; The exoskeleton of insects (cuticle) is an assembly of chitin and cuticle proteins. Its physical properties are determined largely by the proteins it contains, and vary widely with developmental stages and body regions. The genes encoding cuticle proteins are therefore good models to study the molecular mechanisms of signalling by ecdysteroids and juvenile hormones, which regulate molting and metamorphosis in insects. This review summarizes the studies of hormonal regulation of insect cuticle protein genes, and the recent progress in the analysis of the regulatory sequences and transcription factors important for their expression.

Drosophila egg development is an established model for studying epithelial patterning and morphogenesis, but the connection between signaling pathways and egg morphology is still incompletely understood. We have identified a non-classical cadherin, Cad74A, as a putative adhesion gene that bridges epithelial patterning and morphogenesis in the follicle cells. Starting in mid-oogenesis, Cad74A is expressed in the follicle cells that contact the oocyte, including the border cells and most of the columnar follicle cells. However, Cad74A is repressed in two dorsolateral patches of follicle cells, which participate in the formation of tubular respiratory appendages. We show genetically that Cad74A is downstream of the EGFR and BMP signaling pathways and is repressed by the Zn-finger transcription factor Broad. The correlation of Cad74A repression in the cells that bend out of the plane of the follicular epithelium is preserved across Drosophila species and mutant backgrounds exhibiting a range of eggshell phenotypes. Complete removal of Cad74A from the follicle cells causes defects in dorsal appendage formation. Ectopic expression of Cad74A in the roof cells results in shortened, flattened appendages due to the hindered migration of the roof cells. Based on these results, we propose that Cad74A is part of the adhesive machinery that enables robust dorsal appendage formation, and as such provides a link between the patterning of the follicle cells and eggshell morphogenesis.

As chromatin immunoprecipitation (ChIP) sequencing is becoming the dominant technique for studying chromatin modifications, new protocols surface to improve the method. Bioinformatics is also essential to analyze and understand the results, and precise analysis helps us to identify the effects of protocol optimizations. We applied iterative sonication-sending the fragmented DNA after ChIP through additional round(s) of shearing-to a number of samples, testing the effects on different histone marks, aiming to uncover potential benefits of inactive histone marks specifically. We developed an analysis pipeline that utilizes our unique, enrichment-type specific approach to peak calling. With the help of this pipeline, we managed to accurately describe the advantages and disadvantages of the iterative refragmentation technique, and we successfully identified possible fields for its applications, where it enhances the results greatly. In addition to the resonication protocol description, we provide guidelines for peak calling optimization and a freely implementable pipeline for data analysis.

U radu su prikazani rezultati analize zastupljenosti kulturnih sadržaja manjine u okvirnim, izvedbenim i operativnim nastavnim programima talijanskog jezika i prirode i društva za prva četiri razreda talijanske osnovne škole u Hrvatskoj. Ustanovljen je veliki raskorak između zakonskih odredbi i njihove provedbe u formalnom odgojno-obrazovnom sustavu. Broj nastavnih cjelina i tema posvećenih spomenutim sadržajima vrlo je nizak, nedovoljan čak i za djelomično upoznavanje kulturnog bogatstva talijanskog naroda i manjine., The work deals with the results of the analysis of the presence of minority groups’ cultural contents in broad, implemental and operational curricula of Italian language and natural and social science for the first four forms of the Italian primary school in Croatia. There is a big gap between legal acts and their implementation in the formal educational system. The number of teaching units and topics dedicated to previously mentioned contents is very low and insufficient even for the partial understanding of Italian minority group’s cultural heritage.

We present the detection of four rotational emission lines of water vapor, from energy levels Eu/k= 101 - 454 K, in the gravitationally lensed z=3.9 QSO host galaxy APM08279+5255. While the lowest H2O lines are collisionally excited in clumps of warm, dense gas (density of hydrogen nuclei n_H=(3.1 +/- 1.2) x 10^6 cm^-3, gas temperature T_g ~ 105 +/- 21 K), we find that the excitation of the higher lines is dominated by the intense local infrared radiation field. Since only collisionally excited emission contributes to gas cooling, we conclude that H2O is not a significant coolant of the warm molecular gas. Our excitation model requires the radiatively excited gas to be located in an extended region of high 100 micron opacity (tau_100 = 0.9 +/- 0.2). Locally, such extended infrared-opaque regions are found only in the nuclei of ultraluminous infrared galaxies. We propose a model where the infrared-opaque circumnuclear cloud, which is penetrated by the X-ray radiation field of the QSO nucleus, contains clumps of massive star formation where the H2O emission originates. The radiation pressure from the intense local infrared radiation field exceeds the thermal gas pressure by about an order of magnitude, suggesting close to Eddington-limited star formation in these clumps., accepted for publication in Astrophysical Journal Letters; typos fixed