Chitosan-alginate microparticulate delivery system for an alternative route of administration of BCG vaccine

Immunisation against M. tuberculosis with current available BCG vaccine lacks efficacy in preventing adult pulmonary tuberculosis. Targeting nasal mucosa is an attractive option for a more effective immunization. The delivery of BCG via the intranasal route involves overcoming barriers such as crossing the physical barrier imposed by the mucus layer and ciliar remotion, cellular uptake and intracellular trafficking by antigen presenting cells. Due to its biodegradable, immunogenic and mucoadhesive properties, chitosan particulate delivery systems can act both as vaccine carrier and adjuvant, improving the elicited immune response. In this study, different combinations of Chitosan/Alginate/TPP microparticles with BCG were produced as vaccine systems. The developed microparticle system successfully modulates BCG surface physicochemical properties and promotes effective intracellular uptake by human macrophage cell lines Preliminary immune responses were evaluated after s.c. and intranasal immunisation of BALB/c mice. BCG vaccination successfully stimulated the segregation of IgG2a and IgG1, where intranasal immunisation with chitosan/alginate particulate system efficiently elicited a more equilibrated cellular/humoral immune response

Cellular uptake of BCG-loaded chitosan microparticles and in vivo evaluation of immune response following intranasal immunisation

Attenuated Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only currently available vaccine against tuberculosis. It is highly effective in pre-exposure immunisation against TB in children when administered by subcutaneous route to newborns. However, it does not provide permanent protection in adults. In this work, polymeric chitosan-alginate microparticles have been evaluated as potential nasal delivery systems and mucosal adjuvants for live attenuated BCG. Chitosan (CS) has been employed as adjuvant and mucosal permeation-enhancer, and, together with alginate (ALG), as additive to enhance BCG-loaded microparticles (MPs) cellular uptake in a human monocyte cell line, by particle surface modification. The most suitable particles were used for vaccine formulation and evaluation of immune response following intranasal immunisation of BALB/c mice

The Fh8 tag : a fusion partner for simple and cost-effective protein purification in Escherichia coli

Downstream processing is still a major bottleneck in recombinant protein production representing most of its costs. Hence, there is a continuing demand of novel and cost-effective purification processes aiming at the recovery of pure and active target protein. In this work, a novel purification methodology is presented, using the Fh8 solubility enhancer tag as fusion handle. The binding properties of Fh8 tag to a hydrophobic matrix were first studied via hydrophobic interaction chromatography (HIC). The Fh8 tag was then evaluated as a purification handle by its fusion to green fluorescent protein and superoxide dismutase. The purification efficiency of the Fh8-HIC strategy was compared to the immobilized metal ion affinity chromatography (IMAC) using the His6 tag. Results showed that the Fh8-HIC binding mechanism is calcium-dependent in a low salt medium, making the purification process highly selective. Both target proteins were biologically active, even when fused to Fh8, and were successfully purified by HIC, achieving efficiencies identical to those of IMAC. Thus, the Fh8 acts as an effective affinity tag that, together with its previously reported solubility enhancer capability, allows for the design of inexpensive and successful recombinant protein production processes in Escherichia coli.This work was conducted with the financial support of the Fundacao para a Ciencia e Tecnologia (FCT), Portugal, by the fellowship SFRH/BD/46482/2008 (POPH-QREN) to Sofia J. Costa. The FCT project PTDC/CVT/103081/2008 (co-funded by COMPETE) and QREN co-promotion project number 3515 are also acknowledged. The authors thank Dr. Huseyin Besir, Protein Expression and Purification Facility Core, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany, for supplying the pETM11 expression vector, and also Dr. Vitor Costa for kindly providing the SOD target gene used in this work

Impacto de variáveis sociais na resolução de tarefas cognitivas : estudo no início e final do 1º Ciclo do Ensino Básico

Neste texto apresentamos os resultados de um grupo de 60 crianças repartidas pelo 1º e 4º ano de escolaridade, diferenciadas ainda por um meio sócio-económico baixo e rural (crianças de Paredes de Coura) e médio-elevado ou elevado e urbano (crianças a frequentar uma escola particular na cidade de Braga) em provas cognitivas. O estudo pretendeu verificar eventuais discrepâncias na percentagem de acertos e nas bonificações por resolução rápida de itens em alguns dos subtestes da WISC-III (Informação, Compreensão, Completamento de Gravuras, e Cubos) tomando os dois meios sócio-culturais de proveniência das crianças, assim como apreciar se tais discrepâncias diminuíam na passagem das crianças do 1º para o 4º ano de escolaridade (eventual efeito de homogeneização por parte da escola face a outras variáveis sociais de proveniência). Excepto três crianças do meio rural, todas as demais frequentaram o pré-escolar. Os resultados não sugerem uma diferenciação na generalidade dos itens tomando os ratios de acertos aos itens, muito embora essa diferenciação ocorre nas bonificações por execuções mais rápidas das crianças. Esta diferença é favorável às crianças do meio sócio-cultural mais favorecido, aumentando essa discrepância ao passarmos do 1º para o 4º ano de escolaridade. Esta discrepância ocorre com os resultados nos subtestes da WISC-III e nas Matrizes Coloridas Progressivas de Raven, sendo as médias superiores junto das crianças do meio sócio-cultural mais favorecido e, sobretudo, do 4º ano de escolaridade.Fundação para a Ciência e a Tecnologia (FCT)Fundação Calouste Gulbenkian (FCG

Alginate-chitosan particulate delivery systems for mucosal immunization against tuberculosis

Although vaccination is still the most cost-effective strategy for tuberculosis control, there is an urgent need for an improved vaccine. Current BCG vaccine lacks efficacy in preventing adult pulmonary tuberculosis, the most prevalent form of the disease. Targeting nasal mucosa, Mycobacterium tuberculosis infection site, will allow a simpler, less prone to risk of infection and more effective immunization against disease. Due to its biodegradable, immunogenic and mucoadhesive properties, chitosan particulate delivery systems can act both as carrier and as adjuvant, improving the elicited immune response. In this study, BCG was encapsulated in alginate and chitosan microparticles, via a mild ionotropic gelation procedure with sodium tripolyphosphate as a counterion. The particulate system developed shows effective modulation of BCG surface physicochemical properties, suitable for mucosal immunization. Intracellular uptake was confirmed by effective transfection of human macrophage cell lines

Sistema agroflorestal no sul de Minas Gerais: café, frutíferas e madeireiras.

Diante da atual crise ambiental e econômica enfrentada pela agricultura moderna, o presente trabalho teve como objetivos analisar a produtividade, os aspectos econômicos e ambientais de um sistema agroflorestal conduzido em Machado, sul de Minas Gerais há onze anos, tendo como principais produtos o café (Coffea arabica) orgânico certificado pela BCS, que é exportado para outros países, a banana e o eucalipto, que também possuem certificação orgânica e são vendidos no comércio local. A metodologia da pesquisa foi baseada no DRP (Diagnóstico Rural Participativo). Ao contrário dos sistemas convencionais de produção, o sistema agroflorestal estudado evidenciou a viabilidade do modelo de produção nas dimensões produtiva, ecológica e econômica. A baixa produtividade do café no sistema agroflorestal é compensada pela venda do café no mercado internacional, pela produção e comercialização da banana, do eucalipto e obtenção de uma diversidade de alimentos saudáveis para a subsistência das famílias

Aspectos marfanoides e retardo mental ligado ao X associados a anormalidades craniofaciais: síndrome de Lujan-Fryns

Universidade Federal de São Paulo (UNIFESP) Department of NeurologyUNIFESP, Department of NeurologySciEL

On the origin of X-shaped radio galaxies

After a brief, critical review of the leading explanations proposed for the small but important subset of radio galaxies showing an X-shaped morphology (XRGs) we propose a generalized model, based on the jet-shell interaction and spin-flip hypotheses. The most popular scenarios for this intriguing phenomenon invoke either hydrodynamical backflows and over-pressured cocoons or rapid jet reorientations, presumably from the spin-flips of central engines following the mergers of pairs of galaxies, each of which contains a supermassive black hole (SMBH). We confront these models with a number of key observations and thus argue that none of the models is capable of explaining the entire range of salient observational properties of XRGs, although some of the arguments raised in the literature against the spin-flip scenario are probably not tenable. We then propose here a new scenario which also involves galactic mergers but would allow the spin of the central engine to maintain its direction. Motivated by the detailed multi-band observations of the nearest radio galaxy, Centaurus A, this new model emphasizes the role of interactions between the jets and the shells of stars and gas that form and rotate around the merged galaxy and can cause temporary deflections of the jets, occasionally giving rise to an X-shaped radio structure. Although each of the models is likely to be relevant to a subset of XRGs, the bulk of the evidence indicates that most of them are best explained by the jet-shell interaction or spin-flip hypotheses.Comment: 19 pages, major revision including two Appendices and a Table, accepted in Research in Astronomy and Astrophysic