An intact signal peptide on dengue virus E protein enhances immunogenicity for CD8+ T cells and antibody when expressed from modified vaccinia Ankara

Dengue is a global public health concern and this is aggravated by a lack of vaccines or antiviral therapies. Despite the well-known role of CD8(+) T cells in the immunopathogenesis of Dengue virus (DENV), only recent studies have highlighted the importance of this arm of the immune response in protection against the disease. Thus, the majority of DENV vaccine candidates are designed to achieve protective titers of neutralizing antibodies, with less regard for cellular responses. Here, we used a mouse model to investigate CD8(+) T cell and humoral responses to a set of potential DENV vaccines based on recombinant modified vaccinia virus Ankara (rMVA). To enable this study, we identified two CD8(+) T cell epitopes in the DENV-3 E protein in C57BL/6 mice. Using these we found that all the rMVA vaccines elicited DENV-specific CD8(+) T cells that were cytotoxic in vivo and polyfunctional in vitro. Moreover, vaccines expressing the E protein with an intact signal peptide sequence elicited more DENV-specific CD8(+) T cells than those expressing E proteins in the cytoplasm. Significantly, it was these same ER-targeted E protein vaccines that elicited antibody responses. Our results support the further development of rMVA vaccines expressing DENV E proteins and add to the tools available for dengue vaccine development.Parts of this work were supported by the InstitutoNacional de Ciência e Tecnologia de Vacinas–INCTV (National Insti-tute of Science and Technology of Vaccines) and by a FAPEMIGPPM grant (CBB, PPM-00461-11). BRQ was a CAPES/PDSE fellow-ship recipient (8815-11-9). FGF is a CNPq fellowship recipient. DCTis an ARC Future Fellow (FT110100310)

The Role of Chemokines in Mediating Graft Versus Host Disease: Opportunities for Novel Therapeutics

Bone marrow transplantation (BMT) is the current therapy of choice for several malignancies and severe autoimmune diseases. Graft versus host disease (GVHD) is the major complication associated with BMT. T lymphocytes and other leukocytes migrate into target organs during GVHD, become activated and mediate tissue damage. Chemokines are well known inducers of leukocyte trafficking and activation and contribute to the pathogenesis of GVHD. Here, we review the major animal models used to study GVHD and the role of chemokines in mediating tissue damage in these models. The role of these molecules in promoting potential beneficial effects of the graft, especially graft versus leukemia, is also discussed. Finally, the various pharmacological strategies to block the chemokine system or downstream signaling events in the context of GVHD are discussed

Minimal Forbidden Factors of Circular Words

Minimal forbidden factors are a useful tool for investigating properties of words and languages. Two factorial languages are distinct if and only if they have different (antifactorial) sets of minimal forbidden factors. There exist algorithms for computing the minimal forbidden factors of a word, as well as of a regular factorial language. Conversely, Crochemore et al. [IPL, 1998] gave an algorithm that, given the trie recognizing a finite antifactorial language $M$, computes a DFA recognizing the language whose set of minimal forbidden factors is $M$. In the same paper, they showed that the obtained DFA is minimal if the input trie recognizes the minimal forbidden factors of a single word. We generalize this result to the case of a circular word. We discuss several combinatorial properties of the minimal forbidden factors of a circular word. As a byproduct, we obtain a formal definition of the factor automaton of a circular word. Finally, we investigate the case of minimal forbidden factors of the circular Fibonacci words.Comment: To appear in Theoretical Computer Scienc

Caffeic acid loading wound dressing : physicochemical and biological characterization

AIM: Caffeic acid has been described as active against bacteria commonly isolated from wound infections. However, its low stability and poor solubility reduce caffeic acid applicability as a pharmaceutical product. These parameters can be enhanced by complexation with cyclodextrin. The main goal of the present work was to incorporate caffeic acid on cyclodextrin-based hydrogels capable of controlled delivery, in order to be used as antibacterial wound dressing. MATERIALS & METHODS: Cyclodextrin-based hydrogels were prepared by direct crosslinking of -cyclodextrin or hydroxypropyl--cyclodextrin with 1,4-butanediol diglycidyl ether in the presence of hydroxypropyl methylcellulose. RESULTS: The hydrogels obtained combine good physicochemical properties (viscoelasticity, superabsorbency and high ability to retain and deliver caffeic acid) with the preservation of caffeic acid' antibacterial activity and effect on fibroblasts, with gel--cyclodextrin the most suited. CONCLUSION: The hydrogels obtained could be useful as caffeic acid delivery-system devices for the treatment of wound infections.The authors are grateful for the FCT StrategicProject PEst-OE/EQB/LA0023/2013 and the Project 'BioHealth- Biotechnology and Bioengineering approaches to improve healthquality', ref. NORTE-07-0124-FEDER-000027, co-funded by the 'Programa Operacional Regional do Norte' (ON. 2-O Novo Norte), QREN, FEDER. The authors also acknowledge the project 'Consolidating Research Expertise and Resources on Cellular and Molecular Biotechnology at CEB/IBB', ref. FCOMP-01-0124-FEDER-027462. This work is also funded by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE and National Funds through FCT- Foundation for Science and Technology under the project PEst-C/CTM/UI0264/2011. Additionally, the authors would like to thank the FCT for the grant for E Pinho (SFRH/BD/62665/2009).The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed

Punching of reinforced concrete flat slabs – Rational use of high strength concrete

PTDC/ECI-EST/30511/2017; SFRH/BD/76242/2011; The authors are grateful to CIMPOR for providing the cement and Sika for providing VISCOCRETE 20HE and SIKACRETE HD.This paper deals with punching of reinforced high strength concrete (HSC) flat slabs. Despite the use of HSC increased significantly in the last years, the experimental research on punching behavior of HSC slabs is still limited. Furthermore, most of this past research adopted concrete compressive strength lower than 90 MPa. In a previous work by this research group three specimens with concrete compressive strength around 120 MPa and one with normal strength concrete (NSC) were tested. The present work represents the continuation of the previous activity and it is focused on the rational use of HSC. Four specimens with HSC and one of NSC were tested under monotonic vertical loading. The HSC was placed only in the slab-column connection region and it was limited to a thin layer in the compressive zone, in order to have a more economical and sustainable solution. This rational use of the HSC showed excellent results in terms of punching strength. Limiting the HSC to a thin layer in the compressive zone resulted in an almost equal punching strength to that obtained with the slab entirely casted in HSC.authorsversionpublishe

Genetic purity certificate in seeds of hybrid maize using molecular markers.

Um dos principais atributos que conferem à semente uma alta qualidade é sua pureza genética, sendo a principal causa de contaminação em sementes de milho híbrido, a ocorrência de autofecundação do parental feminino. No entanto, ainda não existem metodologias seguras e rápidas para detectar tais contaminações. Desta forma, o presente trabalho foi desenvolvido objetivando certificar a pureza genética em sementes de milho híbrido por meio de diferentes técnicas de marcadores bioquímicos e de DNA. Sementes de dois híbridos e de suas respectivas linhagens, provenientes do programa de melhoramento da Universidade Federal de Lavras foram avaliadas por padrões eletroforéticos das isoenzimas álcool desidrogenase (ADH), esterase (EST), fosfatase ácida (ACP), glutamato-oxalacetato transaminase (GOT), malato desidrogenase (MDH), isocitrato desidrogenase (IDH), 6-fosfoglucamato desidrogenase (PGDH), fosfoglucomutase (PGM), catalase (CAT) e ß-glucosidade (ßGLU) e por marcadores microssatélites. Os sistemas enzimáticos que permitiram separar os híbridos de seus respectivos progenitores foram a catalase, a isocitrato desidrogenase e a esterase, sendo que para a esterase foi observada segregação mendeliana para o híbrido UFLA 8/3, o que torna a certificação da pureza genética mais segura. Os microssatélites além de terem sido eficientes na diferenciação dos híbridos e das linhagens parentais, apresentaram-se como uma técnica de análise rápida, precisa e independente do ambiente. Como o padrão de amplificação dos microssatélites não foi influenciado quando se utilizou o DNA extraído de sementes inteiras ou de folhas, torna-se possível a utilização de sementes como fonte de DNA, o que acelera e facilita a execução das análises de pureza genética. One of the main features that confer high quality to the seed is its genetic purity, in which one of the major causes of contamination is the self-pollination of the female parent. Up to date, there is no accurate and fast methods for detecting such contamination. Thus, this work was carried out to certify the genetic purity in seeds of hybrid maize using different biochemical and DNA-based markers. Two single-cross hybrids and their parental lines derived from the maize breeding program at UFLA were evaluated by isoenzymatic pattern of alcohol dehydrogenase (ADH), esterase (EST), acid phosphatase (ACP), glutamate-oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphoglucomate dehydrogenase (PGDH), catalase (CAT) and ß-glucosidade (ßGLU) and by microsatellites markers. The enzymatic systems that were able to distinguish the hybrids from their parental line were the catalase, the isocitrate dehydrogenase and the esterase. The esterase showed a Mendelian segregation pattern for UFLA 8/3 hybrid, that enables a safer genetic purity certificate. Microsatellites were able to differentiate the hybrid lines and the respective parental lines. Moreover, this technique was fast, precise and without environment effects. For microsatellites, the amplification pattern was identical when young leaves or seeds were used as DNA source. The possibility of using seeds as DNA source would accelerate and facilitate the role process of the genetic purity analysis

Application of a prioritisation scheme for seismic intervention in schools buildings in Italy

A risk management framework has recently been developed to assign priorities for the rehabilitation of school buildings in Italy, and to give timescales within which retrofit or demolition must take place. Since it is not practical to carry out a detailed assessment of the 60,000 Italian state and public schools, the framework is a multiple-level procedure which aims to identify the highest-risk buildings based on filters of increasing detail, and reduces the size of the building inventory at each step. The first risk ranking is based on a strength deficit, which measures the difference between the current design forces defined for the building site and an estimation of the level of seismic resistance which was required at the time of design. The second ranking is based on lateral strength calculations that are already available for a large portion of the Italian masonry building stock, and that are obtained from a survey form that is familiar to Italian engineers. Finally, a simplified displacement-based methodology is used to give a more accurate assessment of seismic risk based on a limited amount of geometrical and material data. The final assessment leads to a capacity ratio and a risk rating, which are used within a transparent procedure to assign priorities for seismic intervention, and timescales within which detailed assessment leading to retrofit or demolition must take place. The first step of the methodology has been applied herein to the school building stock within two Regions in Italy and preliminary results are presented

Amostragem sequencial e marcadores de microssatélites na avaliação da qualidade genética em lotes de sementes de milho.

Edição dos Resumos do XIII Congresso Brasileiro de Sementes, Gramado, RS, set. 2003

Application of a prioritisation scheme for seismic intervention in schools buildings in Italy

A risk management framework has recently been developed to assign priorities for the rehabilitation of school buildings in Italy, and to give timescales within which retrofit or demolition must take place. Since it is not practical to carry out a detailed assessment of the 60,000 Italian state and public schools, the framework is a multiple-level procedure which aims to identify the highest-risk buildings based on filters of increasing detail, and reduces the size of the building inventory at each step. The first risk ranking is based on a strength deficit, which measures the difference between the current design forces defined for the building site and an estimation of the level of seismic resistance which was required at the time of design. The second ranking is based on lateral strength calculations that are already available for a large portion of the Italian masonry building stock, and that are obtained from a survey form that is familiar to Italian engineers. Finally, a simplified displacement-based methodology is used to give a more accurate assessment of seismic risk based on a limited amount of geometrical and material data. The final assessment leads to a capacity ratio and a risk rating, which are used within a transparent procedure to assign priorities for seismic intervention, and timescales within which detailed assessment leading to retrofit or demolition must take place. The first step of the methodology has been applied herein to the school building stock within two Regions in Italy and preliminary results are presented