Expression and characterization of HPV-16 L1 capsid protein in Pichia pastoris

Human papillomaviruses (HPVs) are responsible for the most common human sexually transmitted viral infections. Infection with high-risk HPVs, particularly HPV16, is associated with the development of cervical cancer. The papillomavirus L1 major capsid protein, the basis of the currently marketed vaccines, self-assembles into virus-like particles (VLPs). Here, we describe the expression, purification and characterization of recombinant HPV16 L1 produced by a methylotrophic yeast. A codon-optimized HPV16 L1 gene was cloned into a non-integrative expression vector under the regulation of a methanol-inducible promoter and used to transform competent Pichia pastoris cells. Purification of L1 protein from yeast extracts was performed using heparin-sepharose chromatography, followed by a disassembly/reassembly step. VLPs could be assembled from the purified L1 protein, as demonstrated by electron microscopy. The display of conformational epitopes on the VLPs surface was confirmed by hemagglutination and hemagglutination inhibition assays and by immuno-electron microscopy. This study has implications for the development of an alternative platform for the production of a papillomavirus vaccine that could be provided by public health programs, especially in resource-poor areas, where there is a great demand for low-cost vaccines.

As novas propostas para a seguran??a p??blica no Brasil: uma an??lise comparativa dos projetos Bolsa Forma????o e Mulheres da Paz

A partir do exame qualitativo de dados e informa????es coletadas, lastreado nas teorias do ciclo de pol??ticas p??blicas, esse texto analisa duas recentes iniciativas do governo federal brasileiro na ??rea de seguran??a p??blica ??? os projetos ???Bolsa Forma????o??? e ???Mulheres da Paz???, ambos do Programa Nacional de Seguran??a P??blica com Cidadania (Pronasci). A partir da descri????o e reconstru????o hist??rica desses projetos, tentamos realizar uma compara????o entre alguns de seus principais elementos ou caracter??sticas, concatenando-os com conceitos e abordagens te??ricas, a fim de trazer conclus??es ??teis ?? an??lise e formula????o de pol??ticas p??blicas.

Abastecimento de água e saneamento de águas residuais

O presente relatório diz respeito a um estágio de natureza profissional realizado com o objectivo de constituir trabalho final de mestrado do curso de Especialização em Engenharia Civil – Perfil de Hidráulica, do Instituto Superior de Engenharia de Lisboa. A proposta de estágio, subordinada ao tema “Abastecimento de Água e Saneamento de Águas Residuais” decorreu numa empresa de Consultores de Engenharia (ENGIDRO, Estudos de Engenharia, Lda.) vocacionada para a realização de estudos e projectos na área da Hidráulica, com particular incidência na Hidráulica Urbana e Saneamento Básico. Num gabinete de engenharia existem em geral vários projectos a decorrer em simultâneo, de diferente natureza e grau de complexidade, que se sucedem ao longo do tempo, entregues a diferentes equipas de trabalho. Sendo um Estágio geralmente proposto com vários meses de antecedência relativamente à sua realização, não é possível conhecer à partida quais as equipas de trabalho, ou projectos, sobre os quais vai recair. No presente caso, no âmbito do tema proposto, e de forma a integrar-me uma empresa em pleno funcionamento, iniciei o estágio com a participação em trabalhos pontuais e complementares aos projectos dos Subsistemas de Abastecimento de Água da Terra Fria Transmontana – Subsistema do Alto Sabor, para as Águas de Trás-os-Montes e Alto Douro, concretamente na elaboração de mapas de medições e orçamentação dos trabalhos projectados. Concluída essa primeira intervenção, desenvolvi os trabalhos de concepção e dimensionamento, a nível de Projecto de Execução, das Redes de Abastecimento de Água e de Drenagem de Águas Residuais do Estaleiro Definitivo da obra do Aproveitamento Hidroeléctrico do Baixo Sabor, trabalhos realizados com o apoio e coordenação do Prof. Eng. Luís Mendes. Por último, integrado numa equipa multidisciplinar constituída por técnicos de várias empresas ligadas em consórcio, e enquadrado pela Eng.ª Paula Almeida, pelo Prof. Eng. Alexandre Borga e pelo Prof. Eng. Luís Mendes, acompanhei os Estudos e Projectos da Frente de Drenagem Cais do Sodré-Alcântara – Interceptor Cais do Sodré – Alcântara II. Com a orientação do Prof. Eng. Luís Mendes, desenvolvi todos estes trabalhos de uma forma proveitosa, sempre com o auxílio de uma equipa formada em Hidráulica em plena actividade. Neste período usei a minha experiência profissional adquirida como Director de Obra na elaboração dos cálculos, elaboração das medições e conferência de desenhos.

Assessing the conservation status of the strict endemic Desertas wolf spider, Hogna ingens (Araneae, Lycosidae)

The Desertas Islands (Madeira, Portugal) are the sole home of one of the largest and rarest wolf spiderspecies, Hogna ingens (Blackwall 1857) (Araneae, Lycosidae). Despite its size, it inhabits a single valleyin the North of the Deserta Grande Island, Vale da Castanheira, currently invaded by the herb Phalarisaquatica. This invasive species competes with the native flora and was subject to several eradicationexperiments, namely through fire and chemicals. The objectives of this work were to: (1) estimate thecurrent distribution and abundance of H. ingens and respective trends; (2) evaluate the impact of theinvasive plant and eradication methods on the spider population; (3) suggest future measures for therecovery of the species; and (4) evaluate its conservation status according to the IUCN criteria. The current distribution of H. ingens covers 23 ha, a recent reduction from its original 83 ha, correspond-ing to the entire Vale da Castanheira. A total of 4447 and 4086 adults and 71,832 and 24,635 juvenileswere estimated to live in the valley during 2011 and 2012, respectively. We found a significant negativeimpact of P. aquatica cover on the presence and abundance of H. ingens and that chemical treatmentspecifically directed towards the invasive plant species may be the only way to effectively recover thespider's habitat. We suggest (1) regular monitoring; (2) extend chemical treatments; (3) ex-situ conserva-tion with future reintroduction of adults. Based on the current area of occupancy (AOO) of H. ingens and itsrecent decline in both AOO and number of individuals, it was recently classified as Critically Endangeredby IUCN and we suggest its urgent inclusion in the Habitats Directive species lists.

Como efetuar multiplicações sem saber a tabuada?

Apresenta-se uma biografia resumida de John Napier e a descrição do dispositivo "Ossos de Napier". São ainda ilustradas e explicadas multiplicações recorrendo aos "Ossos de Napier".

RiSeG: a ring based secure group communication protocol for resource-constrained wireless sensor networks

Securing group communication in wireless sensor networks has recently been extensively investigated. Many works have addressed this issue, and they have considered the grouping concept differently. In this paper, we consider a group as being a set of nodes sensing the same data type, and we alternatively propose an efficient secure group communication scheme guaranteeing secure group management and secure group key distribution. The proposed scheme (RiSeG) is based on a logical ring architecture, which permits to alleviate the group controller’s task in updating the group key. The proposed scheme also provides backward and forward secrecy, addresses the node compromise attack, and gives a solution to detect and eliminate the compromised nodes. The security analysis and performance evaluation show that the proposed scheme is secure, highly efficient, and lightweight. A comparison with the logical key hierarchy is preformed to prove the rekeying process efficiency of RiSeG. Finally, we present the implementation details of RiSeG on top of TelosB sensor nodes to demonstrate its feasibility.

Performance of concrete made with aggregates recycled from precasting industry waste: influence of the crushing process

The aim of this paper is to evaluate the influence of the crushing process used to obtain recycled concrete aggregates on the performance of concrete made with those aggregates. Two crushing methods were considered: primary crushing, using a jaw crusher, and primary plus secondary crushing (PSC), using a jaw crusher followed by a hammer mill. Besides natural aggregates (NA), these two processes were also used to crush three types of concrete made in laboratory (L20, L45 e L65) and three more others from the precast industry (P20, P45 e P65). The coarse natural aggregates were totally replaced by coarse recycled concrete aggregates. The recycled aggregates concrete mixes were compared with reference concrete mixes made using only NA, and the following properties related to the mechanical and durability performance were tested: compressive strength; splitting tensile strength; modulus of elasticity; carbonation resistance; chloride penetration resistance; water absorption by capillarity; water absorption by immersion; and shrinkage. The results show that the PSC process leads to better performances, especially in the durability properties. © 2014 RILEM

Performance of concrete made with aggregates recycled from precasting industry waste: influence of the crushing process

The aim of this paper is to evaluate the influence of the crushing process used to obtain recycled concrete aggregates on the performance of concrete made with those aggregates. Two crushing methods were considered: primary crushing, using a jaw crusher, and primary plus secondary crushing (PSC), using a jaw crusher followed by a hammer mill. Besides natural aggregates (NA), these two processes were also used to crush three types of concrete made in laboratory (L20, L45 e L65) and three more others from the precast industry (P20, P45 e P65). The coarse natural aggregates were totally replaced by coarse recycled concrete aggregates. The recycled aggregates concrete mixes were compared with reference concrete mixes made using only NA, and the following properties related to the mechanical and durability performance were tested: compressive strength; splitting tensile strength; modulus of elasticity; carbonation resistance; chloride penetration resistance; water absorption by capillarity; water absorption by immersion; and shrinkage. The results show that the PSC process leads to better performances, especially in the durability properties.

Durability performance of self-compacting concrete (SCC) with binary and ternary mixes of fly ash and limestone filler

The basic objective of this work is to evaluate the durability of self-compacting concrete (SCC) produced in binary and ternary mixes using fly ash (FA) and limestone filler (LF) as partial replacement of cement. The main characteristics that set SCC apart from conventional concrete (fundamentally its fresh state behaviour) essentially depend on the greater or lesser content of various constituents, namely: greater mortar volume (more ultrafine material in the form of cement and mineral additions); proper control of the maximum size of the coarse aggregate; use of admixtures such as superplasticizers. Significant amounts of mineral additions are thus incorporated to partially replace cement, in order to improve the workability of the concrete. These mineral additions necessarily affect the concrete’s microstructure and its durability. Therefore, notwithstanding the many well-documented and acknowledged advantages of SCC, a better understanding its behaviour is still required, in particular when its composition includes significant amounts of mineral additions. An ambitious working plan was devised: first, the SCC’s microstructure was studied and characterized and afterwards the main transport and degradation mechanisms of the SCC produced were studied and characterized by means of SEM image analysis, chloride migration, electrical resistivity, and carbonation tests. It was then possible to draw conclusions about the SCC’s durability. The properties studied are strongly affected by the type and content of the additions. Also, the use of ternary mixes proved to be extremely favourable, confirming the expected beneficial effect of the synergy between LF and FA. © 2015 RILEM.

Cation transporters/channels in plants: Tools for nutrient biofortification

Cation transporters/channels are key players in a wide range of physiological functions in plants, including cell signaling, osmoregulation, plant nutrition and metal tolerance. The recent identification of genes encoding some of these transport systems has allowed new studies toward further understanding of their integrated roles in plant. This review summarizes recent discoveries regarding the function and regulation of the multiple systems involved in cation transport in plant cells. The role of membrane transport in the uptake, distribution and accumulation of cations in plant tissues, cell types and subcellular compartments is described. We also discuss how the knowledge of inter- and intra-species variation in cation uptake, transport and accumulation as well as the molecular mechanisms responsible for these processes can be used to increase nutrient phytoavailability and nutrients accumulation in the edible tissues of plants. The main trends for future research in the field of biofortification are proposed.

Asialotransferrina Discriminante do Consumo Excessivo Activo de Álcool, Sub-Agudo e Crónico. Utilidade Clínica na Doença Hepática

A determinação dos níveis séricos de transferrina deficiente em carbo-hidratos (CDT) e do rácio de transferrina no consumidor abusivo persistente de álcool aparece com promissora utilidade na área da alcoologia. Na série actual demonstra-se excelente especificidade (970 o) no entanto com fraca sensibilidade (52° o) para o CDT. Contudo o rácio CDT Tft adiciona-lhe uma sensibilidade acrescida atingindo 740 o mantendo a elevada especificidade. Nos consumidores abusivos 70g dia) persistentes, este índice que se correlaciona positivamente com a transferrina sérica, é capaz de escurtinar em elevada frequência estas capitações de álcool e mostra uma informação independente pois não se correlaciona significativamente com os níveis dos marcadores biológicos tradicionais (AST, ALT, GGT, VGM). Embora com limitações metodológicas definidas, estes índices prenunciam, com melhoria da acessibilidade técnica, uma aplicabilidade prática no rastreio de consumidores abusivos crónicos. Na área da hepatologia o comportamento do CDT e ácio da transferrina é capaz de afirmar em elevada frequência o envolvimento do etanol no estudo da natureza de uma doença hepática crónica. No entanto, o grau de lesão do figado tomado pelo índice hepático PGA, não influencia apreciavelmente os níveis séricos do CDT e rácio de transferrina. Na presente série, as circunstancias e condições do consumo de álcool parecem ser o determinante independente do carácter informativo que estes índices revelam.

Design and validation of a biomechanical bioreactor for cartilage tissue culture

Specific tissues, such as cartilage undergo mechanical solicitation under their normal performance in human body. In this sense, it seems necessary that proper tissue engineering strategies of these tissues should incorporate mechanical solicitations during cell culture, in order to properly evaluate the influence of the mechanical stimulus. This work reports on a user-friendly bioreactor suitable for applying controlled mechanical stimulation - amplitude and frequency - to three dimensional scaffolds. Its design and main components are described, as well as its operation characteristics. The modular design allows easy cleaning and operating under laminar hood. Different protocols for the sterilization of the hermetic enclosure are tested and ensure lack of observable contaminations, complying with the requirements to be used for cell culture. The cell viability study was performed with KUM5 cells.

Characterization of dry-stack interlocking compressed earth blocks

Earth has been a traditional building material to construct houses in Africa. One of the most common techniques is the use of sun dried or kiln fired adobe bricks with mud mortar. Fired bricks are the main cause for deforestation in countries like Malawi. Although this technique is low-cost, the bricks vary largely in shape, strength and durability. This leads to weak houses which suffer considerable damage during floods and seismic events. One solution is the use of dry-stack masonry with stabilized interlocking compressed earth blocks (ICEB). This technology has the potential of substituting the current bricks by a more sustainable kind of block. This study was made in the context of the HiLoTec project, which focuses on houses in rural areas of developing countries. For this study, Malawi was chosen for a case study. This paper presents the experimental results of tests made with dry-stack ICEBs. Soil samples from Malawi were taken and studied. Since the experimental campaign could not be carried out in Malawi, a homogenization process of Portuguese soil was made to produce ICEBs at the University of Minho, Portugal. Then, the compression and tensile strength of the materials was determined via small cylinder samples. Subsequently, the compression and flexural strength of units were determined. Finally, tests to determine the compressive strength of both prisms and masonry wallets and to determine the initial shear strength of the dry interfaces were carried out. This work provides valuable data for low-cost eco-efficient housing

Hortus Indicus Malabaricus :continens regni Malabarici apud Indos cereberrimi onmis generis plantas rariores, Latinas, Malabaricis, Arabicis, Brachmanum charactareibus hominibusque expressas ... /adornatus per Henricum van Rheede, van Draakenstein, ... et Johannem Casearium ... ; notis adauxit, & commentariis illustravit Arnoldus Syen ...

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