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.

Natural selection and quantitative genetics of life-history traits in Western women: A twin study

Whether contemporary human populations are still evolving as a result of natural selection has been hotly debated. For natural selection to cause evolutionary change in a trait, variation in the trait must be correlated with fitness and be genetically heritable and there must be no genetic constraints to evolution. These conditions have rarely been tested in human populations. In this study, data from a large twin cohort were used to assess whether selection Will cause a change among women in contemporary Western population for three life-history traits: age at menarche, age at first reproduction, and age at menopause. We control for temporal variation in fecundity (the baby boom phenomenon) and differences between women in educational background and religious affiliation. University-educated women have 35% lower fitness than those with less than seven years education, and Roman Catholic women have about 20% higher fitness than those of other religions. Although these differences were significant, education and religion only accounted for 2% and 1% of variance in fitness, respectively. Using structural equation modeling, we reveal significant genetic influences for all three life-history traits, with heritability estimates of 0.50, 0.23, and 0.45, respectively. However, strong genetic covariation with reproductive fitness could only be demonstrated for age at first reproduction, with much weaker covariation for age at menopause and no significant covariation for age at menarche. Selection may, therefore, lead to the evolution of earlier age at first reproduction in this population. We also estimate substantial heritable variation in fitness itself, with approximately 39% of the variance attributable to additive genetic effects, the remainder consisting of unique environmental effects and small effects from education and religion. We discuss mechanisms that could be maintaining such a high heritability for fitness. Most likely is that selection is now acting on different traits from which it did in pre-industrial human populations.

Stem cells in the aetiopathogenesis and therapy of rheumatic disease

Animal models of autoimmune disease and case reports of patients with these diseases who have been involved in bone marrow transplants have provided important data implicating the haemopoietic stem cell in rheumatic disease pathogenesis. Animal and human examples exist for both cure and transfer of rheumatoid arthritis, systemic lupus erythematosus (SLE) and other organ-specific diseases using allogeneic haemopoietic stem cell transplantation. This would suggest that the stem cell in these diseases is abnormal and could be cured by replacement of a normal stem cell although more in vitro data are required in this area. Given the morbidity and increased mortality in some patients with severe autoimmune diseases and the increasing safety of autologous haemopoietic stem cell transplantation (HSCT), pilot studies have been conducted using HSCT in rheumatic diseases. It is still unclear whether an autologous graft will cure these diseases but significant remissions have been obtained which have provided important data for the design of randomized trials of HSCT versus more conventional therapy. Several trials are now open to accrual under the auspices of the European Bone Marrow Transplant Group/European League Against Rheumatism (EBMT/EULAR) registry. Future clinical and laboratory research will need to document the abnormalities of the stem cell of a rheumatic patient because new therapies based on gene therapy or stem cell differentiation could be apllied to these diseases. With increasing safety of allogeneic HSCT it is not unreasonable to predict cure of some rheumatic diseases in the near future.

Stability of nest range, home range and movement of the northern bettong (Bettongia tropica) following moderate-intensity fire in a tropical woodland, north-eastern Queensland

Nest use, home-range characteristics and nightly movements by the northern bettong (Bettongia tropica) were examined before and after a low- to moderate-intensity fire in sclerophyll woodland in north-eastern Australia using radio-telemetry. In all, 23 animals were radio-tracked at three-month intervals between February 1995 and May 1996. During November 1995 a low- intensity experimental fire burned the entire home range of most animals. The northern bettong appeared fairly catholic in choice of nest site, with a variety of nest locations and nesting materials used. Prior to the fire, nests were generally located in areas of dense cover, such as the skirts of grass trees (46%) or grass close to a log (29%). After fire removed most ground cover in the nesting areas of most animals, bettongs used remaining shelter such as boulder piles (45%), recently fallen trees (8%) and patches of unburnt vegetation (21%). Nest areas (10.1 ha) of males were significantly larger than those of females (5.4 ha). Home ranges of both sexes were large (59 ha) and most ranges lacked distinct core areas, suggesting that bettongs used all parts of their home ranges equally. High mean rates of nightly movement by the northern bettong indicated that large distances were moved within home ranges during nightly foraging. No significant fire-related changes were detected in home-range size, home-range location, nest-area location or mean rates of nightly movement, suggesting that the northern bettong is well adapted to the low- and medium-intensity fires that characterise its habitat.

Disability focus groups: A strategy for including people with disability in parishes

This paper reports the findings of an exploratory qualitative study examining parish awareness of the need to include individuals with disability in genuine and meaningful ways. Based on the interviews of over thirty persons, including people with disability, parents, parish workers, volunteers and human service workers within church agencies in the Catholic Archdiocese of Brisbane, Australia, the authors devised a process for including individuals in parishes. This process, known as a Disability Focus Group, was then implemented in one parish as a pilot study. The outcomes of the pilot are discussed.

Severity of sports-related concussion and neuropsychological test performance

Concussion severity grades according to the Cantu, Colorado Medical Society, and American Academy of Neurology systems were not clearly related to the presence or duration of impaired neuropsychological test performance in 21 professional rugby league athletes. The use of concussion severity guidelines and neuropsychological testing to assist return to play decisions requires further investigation.

Three poverties

This paper is a discourse on ideological communication on poverty in Latin America from three different perspectives: the Catholic priest, the Shining Path guerrilla and the development economist. The paper concludes with a discussion of these three perspectives from the perspective of the theory of communicative action espoused by Jurgen Habermas.

Applied physiology and game analysis of rugby union

Increased professionalism in rugby has elicited rapid changes in the fitness profile of elite players. Recent research, focusing on the physiological and anthropometrical characteristics of rugby players, and the demands of competition are reviewed. The paucity of research on contemporary elite rugby players is highlighted, along with the need for standardised testing protocols. Recent data reinforce the pronounced differences in the anthropometric and physical characteristics of the forwards and backs. Forwards are typically heavier, taller, and have a greater proportion of body fat than backs. These characteristics are changing, with forwards developing greater total mass and higher muscularity. The forwards demonstrate superior absolute aerobic and anaerobic power, and Muscular strength. Results favour the backs when body mass is taken into account. The scaling of results to body mass can be problematic and future investigations should present results using power function ratios. Recommended tests for elite players include body mass and skinfolds, vertical jump, speed, and the multi-stage shuttle run. Repeat sprint testing is a possible avenue for more specific evaluation of players. During competition, high-intensity efforts are often followed by periods of incomplete recovery. The total work over the duration of a game is lower in the backs compared with the forwards; forwards spend greater time in physical contact with the opposition while the backs spend more time in free running, allowing them to cover greater distances. The intense efforts undertaken by rugby players place considerable stress on anaerobic energy sources, while the aerobic system provides energy during repeated efforts and for recovery. Training should focus on repeated brief high-intensity efforts with short rest intervals to condition players to the demands of the game. Training for the forwards should emphasise the higher work rates of the game, while extended rest periods can be provided to the backs. Players should not only be prepared for the demands of competition, but also the stress of travel and extreme environmental conditions. The greater professionalism of rugby union has increased scientific research in the sport; however, there is scope for significant refinement of investigations on the physiological demands of the game, and sports-specific testing procedures.

Portaria n?? 81, de 11 de maio de 2011

A pol??tica de acesso ?? internet a ser observada pelos servidores, colaboradores, alunos e visitantes e usu??rios do servi??o de acesso ?? internet disponibilizado pela ENAP foi aprovada pela Portaria n?? 81, de 11 de maio de 2011.

Resolu????o n?? 05, de 25 de abril de 2013: disp??e sobre as regras e procedimentos para a concess??o, parcelamento e pagamento da remunera????o de f??rias dos servidores da ENAP

A Resolu????o n?? 5, de 25 de abril de 2013 disp??e sobre as regras e procedimentos para a concess??o, parcelamento e pagamento da remunera????o de f??rias dos servidores da ENAP.

As ideias pedagógicas no desenvolvimento da educação na paraíba (1964-1985): um estudo sobre a influência da pedagogia católica

As ideias pedagógicas no desenvolvimento da educação na Paraíba (1964-1985): um estudo sobre a influência da pedagogia católica

O papel da Organização das Nações Unidas em Angola

RESUMO: Angola tem sido ao longo dos tempos objecto de cobiça por parte de países estrangeiros por motivações diversas, que foram desde interesses coloniais – como aconteceu com Portugal, que durante séculos a colonizou, e da Holanda, que dominou Luanda entre 1641 e 1648 – ou por interesses hegemónicos, ideológicos e políticos – como sucedeu com os EUA e a URSS, mas também com a Republica Popular da China e Cuba – e até por interesses regionais – como foram os casos da África do Sul e dos contíguos Congo Belga, hoje Republica Democrática do Congo, e Zâmbia. No entanto, a todos estes interesses diversos não se pode excluir um que lhes é transversal: o interesse económico. De facto, os 1246700 km² de que Angola dispõe, aliados à sua excelente localização geográfica com uma extensa costa atlântica e a sua extraordinária riqueza em recursos naturais podem explicar este envolvimento estrangeiro na História de Angola. No que diz respeito ao objecto da Dissertação, o envolvimento da ONU nas questões relacionadas com Angola remonta à década de 50, ou seja, ao período colonial e muito antes da independência do país em 1975, devido à política descolonizadora saída da II Guerra Mundial. Além disso, a dinâmica que emergiu da II Guerra Mundial, rapidamente, reconfigurou o panorama político internacional em dois blocos: o ocidental liderado pelos EUA e o de Leste liderado pela URSS, que se envolveram numa Guerra Fria, polarização cujos efeitos se ligam de forma trágica à guerra em Angola, primeiro para a autodeterminação, e, depois, já num contexto de independência, num conflito armado que ultrapassou o plano interno. Os interesses dos EUA e da URSS, que começaram por ser antagónicos, deram lugar em 1989 com o fim da bipolaridade a uma cooperação mais aberta e uma abertura política em Angola rumo à paz e ao início da construção da democracia. Neste trabalho estuda-se o papel da ONU em Angola, quer no período de luta pela independência, quer depois, na busca da paz no sangrento conflito – nem sempre civil – que mesmo antes da data da independência, a 11 de Novembro de 1975, e até Fevereiro de 2002, dilacerou o país. Procura-se, igualmente, analisar o contributo da ONU na consolidação das instituições e na construção de um regime democrático em Angola. ABSTRACT: Angola has been along time subject to the greed of several foreign countries for many reasons and motivations which go from colonial interests - that is the cases of, Portugal which for centuries colonized it, and Netherlands under whose administration had been Luanda between 1641 and 1648 - or for hegemonic, ideological and political interests - as it happened in regard to USA and USSR, but also People‟s Republic of China, Cuba, - and even for regional interests - regarding South Africa Republic, and the neighbouring countries, Democratic Republic of Congo (ancient Republic of the Congo “Leopoldville”), and the Republic of Zambia. On the other hand to these interests we may join another which is transversal to all of them: economic interest. Effectively, Angola‟s 481,351 square miles (1,246,700 Km2) estimated area, combined with its excellent geographical location with a lengthy Atlantic coast, its extraordinary richness in natural resources may well explain this foreign participation in its Political History. Concerning the objective of this work, the UN has been involved in matters regarding Angola since the decade of 50 of the last century, during the colonial period, long before the independence of the country in 1975, due to the decolonization policy emerged from the Second World War. Furthermore, after the Second World War, international environment has changed, transforming quickly the world into two main blocs, the West with the leadership of the USA and the East with the leadership of USSR which went into a Cold War. The effects of this polarization reached tragically Angola, early in the fight for self-determination, and went on later after independence in an armed conflict, which has overcome the internal dimension. The USA and USSR interests, at the beginning being antagonistic had become by 1989, with the end of bipolarity, more cooperative, leading Angola to a political reform towards peace and beginning the construction of democracy. In this academic work it‟s studied the UNO role in Angola since the fight for self-determination early in the sixties of last century, and later in the search for peace during the bloody - and not always civil – war conflict which very before independence date in 11th November 1975, and as long as 2002, divided the country. Additionally, this work aims to understand the UNO contribution to consolidate institutions and to promote democracy in Angola.

Superprodução, purificação e caracterização parcial de complexos proteína-polissacáridos inovadores a partir de estirpes de basidiomicetos

O presente trabalho teve como objectivo o estudo de meios de cultura, tendo em vista a superprodução de complexos proteína-polissacáridos, intracelulares e extracelulares, a partir dos basidiomicetos Pleurotus ostreatus e Lentinula edodes. O meio de cultura suplementado com soro de leite apresentou diversas vantagens para a obtenção dessas macromoléculas biológicas, face aos restantes meios analisados, pelo que foi seleccionado para a produção em fermentador de laboratório em regime descontínuo. O crescimento da cultura foi efectuado a 28°C, 200 rpm e durante 10 dias, e as produtividades de complexos proteína-polissacáridos obtidas para o Pleurotus ostreatus foram de 0,008 ± 8,12 x 10-5 g/(L.dia) e de 0,324 ± 0,005 g/(L.dia), para os complexos precipitados a partir da biomassa e do caldo de fermentação, respectivamente. Posteriormente, os complexos intracelulares e extracelulares foram purificados por cromatografia de filtração em gel e parcialmente caracterizados, revelando concentrações de polissacáridos superiores à concentração proteica, pseudo-actividade de superóxido dismutase, e valores de Mr entre 64 e 9 000kDa. As amostras de complexos proteína-polissacáridos foram caracterizadas por HPLC e apresentaram picos de UV e IR com tempos de retenção de aproximadamente 6 e 12 minutos, respectivamente. Após hidrólise ácida, os complexos heteropolissacáridos obtidos apresentaram na sua constituição glucose, ramnose e arabinose. Adicionalmente, a análise dos complexos proteína-polissacáridos por FT-IR revelou bandas de absorção características destas macromoleculas biológicas, designadamente, a 846,5; 1032,3; 1186,4; 1471,0; 1648,8; 2739,6 e 3419,3 cm-1 para o Pleurotus ostreatus e 862,3; 1044,1; 1474,9; 1644,8; 2372,0; 2992,5; e 3415,4 cm-1 para Lentinula edodes.