High circulating levels of the dengue virus nonstructural protein NS1 early in dengue illness correlate with the development of dengue hemorrhagic fever

Infection with any 1 of 4 dengue viruses produces a spectrum of clinical illness ranging from a mild undifferentiated febrile illness to dengue fever (DF) to dengue hemorrhagic fever (DHF), a potentially life-threatening disease. The morbidity and mortality of DHF can be reduced by early hospitalization and careful supportive care. To determine its usefulness as a predictor of DHF, plasma levels of the secreted dengue virus nonstructural protein NS1 (sNS1) were measured daily in 32 children with dengue-2 virus infections participating in a prospective, hospital-based study. Free sNS1 levels in plasma correlated with viremia levels and were higher in patients with DHF than in those with DF. An elevated free sNS1 level (greater than or equal to600 ng/mL) within 72 h of illness onset identified patients at risk for developing DHF.

Catalytically active Dengue virus NS3 protease forms aggregates that are separable by size exclusion chromatography

An active form of the Dengue virus protease NS3 (CF40.Gly.NS3pro) was expressed in Escherichia coli. This construct consists of a critical 40 amino acid cofactor domain from NS2B fused to the N-terminal 184 amino acid protease domain of NS3 via a flexible, covalent linker (Gly(4)SerGly(4)). The recombinantly produced protein is soluble and has a hexa-histidine tag engineered at the N-terminus for ease of purification using metal affinity chromatography. However, the presence of lower molecular weight impurities after affinity chromatography indicated the need for additional purification steps. The consistent appearance of these impurities suggested that they may be the products of proteolysis and/or auto-proteolysis. The latter possibility was subsequently excluded by the observation of the same impurities in a purified, catalytically inactive form of the recombinant protease (CF40.Gly.NS3pro.SA). Further analysis indicated that these impurities may represent premature translation termination products. Regardless of their origin, they were shown to form various sized aggregates with full-length CF40.Gly.NS3pro that can be separated by size exclusion chromatography, yielding fractions of active protease of sufficient purity for crystallisation trials. The ultimate goal of these studies is to obtain a crystal structure of a catalytically active form of the Dengue virus NS3 protease for structure-based drug design. (C) 2002 Elsevier Science (USA). All rights reserved.

Production of the baculovirus-expressed dengue virus glycoprotein NS1 can be improved dramatically with optimised regimes for fed-batch cultures and the addition of the insect moulting hormone, 20-Hydroxyecdysone

A perennial problem in recombinant protein expression is low yield of the product of interest. A strategy which has been shown to increase the production of baculovirus-expressed proteins is to utilise fed-batch cultures. One disadvantage of this approach is the time-consuming task of optimising the feeding strategy. Previously, a statistical optimisation routine was applied to develop a feeding strategy that increased the yield of beta-Galactosidase (beta-Gal) by 2.4-fold (Biotechnol. Bioeng, 59 (1998) 178). This involves the single addition of nutrient concentrates (amino acids, lipids. glucose and yeastolate ultrafiltrate) into Sf9 cell cultures grown in SF900II medium. In this study, it is demonstrated that this optimised fed-batch strategy developed for a high-yielding intracellular product beta-Gal could be applied successfully to a relatively low-yielding glycosylated and secreted product such as the dengue virus glycoprotein NS1. Optimised batch infections yielded 4 mug/ml of NS1 at a peak cell density of 4.2 x 10(6) cells/ml. In contrast. optimised fed-batch infections exhibited a 3-fold improvement in yield, with 12 mug ml of NS1 produced at a peak cell density of 11.3 x 10(6) cells/ml. No further improvements in yield were recorded when the feed volumes were doubled and the peak cell density was increased to 23 x 10(6) cells/ml, unless the cultures were stimulated by the addition of 4 mug/ml of 20-Hydroxyecdysone (an insect moulting hormone). In this case, the NS1 yield was increased to 20 mug/ml. which was nearly 5-fold higher than optimised batch cultures. (C) 2002 Elsevier Science B.V. All rights reserved.

Control of Aedes vectors of dengue in three provinces of Vietnam by use of Mesocyclops (Copepoda) and community-based methods validated by entomologic, clinical, and serological surveillance

We describe remarkable success in controlling dengue vectors, Aedes aegypti (L.) and Aedes albopictus (Skuse), in 6 communes with 11,675 households and 49,647 people in the northern provinces of Haiphong, Hung Yen, and Nam Dinh in Vietnam. The communes were selected for high-frequency use of large outdoor concrete tanks and wells. These were found to be the source of 49.6-98.4% of Ae. aegypti larvae, which were amenable to treatment with local Mesocyclops, mainly M. woutersi Van der Velde, M. aspericornis (Daday) and M. thermocyclopoides Harada. Knowledge, attitude, and practice surveys were performed to determine whether the communities viewed dengue and dengue hemorrhagic fever as a serious health threat; to determine their knowledge of the etiology, attitudes, and practices regarding control methods including Mesocyclops; and to determine their receptivity to various information methods. On the basis of the knowledge, attitude, and practice data, the community-based dengue control program comprised a system of local leaders, health volunteer teachers, and schoolchildren, supported by health professionals. Recycling of discards for economic gain was enhanced, where appropriate, and this, plus 37 clean-up campaigns, removed small containers unsuitable for Mesocyclops treatment. A previously successful eradication at Phan Boi village (Hung Yen province) was extended to 7 other villages forming Di Su commune (1,750 households) in the current study. Complete control was also achieved in Nghia Hiep (Hung Yen province) and in Xuan Phong (Nam Dinh province); control efficacy was greater than or equal to 99.7% in the other 3 communes (Lac Vien in Haiphong, Nghia Dong, and Xuan Kien in Nam Dinh). Although tanks and wells were the key container types of Ae. aegypti productivity, discarded materials were the source of 51% of the standing crop of Ae. albopictus. Aedes albopictus larvae were eliminated from the 3 Nam Dinh communes, and 86-98% control was achieved in the other 3 communes. Variable dengue attack rates made the clinical and serological comparison of control and untreated communes problematic, but these data indicate that clinical surveillance by itself is inadequate to monitor dengue transmission.

Epidemiological significance of subterranean Aedes aegypti (Diptera : Culicidae) breeding sites to dengue virus infection in charters towers, 1993

The objective of this study was to determine the epidemiological significance of subterranean mosquito breeding sites to the 1993 outbreak of dengue fever (type 2) in the northern Queensland town of Charters Towers, Australia. In recent studies on subterranean mosquito breeding, containers such as wells and service manholes have been shown to be important breeding sites to Australia's only dengue vector, Aedes aegypti (L.). This study demonstrates a direct epidemiological association between subterranean breeding sites and dengue virus infection. The mean distance between residents seropositive for dengue 2 and the nearest subterranean container (113 m) was significantly less than for a randomly selected control (191 m), (F = 81.9; df = 1, 478; P < 0.001). Residents positive for dengue 2 antibodies was 2.47 (95% confidence interval 1.88-3.24) times higher for those living within 160 m of a well or service manhole, compared with those residing further away. These findings emphasize the importance of including subterranean water containers in Ae. aegypti surveillance and control programs.

Enhanced vector competence of Aedes aegypti (Diptera : Culicidae) from the torres strait compared with mainland Australia for dengue 2 and 4 viruses

Australian Aedes aegypti (L.) mosquitoes colonized from the Torres Strait and three mainland localities (Charters Towers, Townsville, and Cairns) were fed on blood suspensions containing dengue virus type 2 (DEN-2) or dengue virus type 4 (DEN-4). Variation was found in oral susceptibility to DEN-2 (59-99% infection) and DEN-4 (28-79% infection) among Ae. aegypti assayed for virus at 8, 12, 16, or 20 d after ingestion of infected blood. Torres Strait Ae. aegypti were the most susceptible to DEN-2 and were significantly more efficient in transmission to capillary tube at 16 d (76% transmission) than mainland Ae. aegypti populations (20-28% transmission). Torres Strait Ae. aegypti were also the most susceptible to DEN-4, although transmission did not vary significantly from mainland populations at 16 d (12% compared with 0-4%) or 20 d (16% compared with 4-16%). Disseminated infection (i.e., leg infection) with either DEN-2 or DEN-4 was not an accurate predictor of transmission potential. This study demonstrates differences among Australian Ae. aegypti populations in vector competence for DEN-2 and DEN-4. Torres Strait Ae. aegypti were more frequently infected and able to transmit DEN-2 at higher rates than mainland populations. These data indicate that the Torres Strait region is potentially more receptive to dengue transmission than mainland localities, a finding discussed with respect to past outbreaks.

O combate ?? viol??ncia contra a mulher: a luta entre antigos valores e novos padr??es de pol??ticas p??blicas

O caso relata uma situa????o de viol??ncia dom??stica vivenciada por uma mulher negra e os obst??culos por ela enfrentados para denunciar o agressor e fazer valer seus direitos. Mostra a contradi????o entre normas e sua efetiva aplica????o, quando o comportamento de agentes p??blicos ainda conserva padr??es e valores de um Estado autorit??rio, patriarcal e escravocrata. O caso ?? fict??cio, mas espelha situa????es reais coletadas em documentos e relat??rios da Ouvidoria da Secretaria de Pol??ticas para as Mulheres- SPM/PR. O estudo suscita discuss??es sobre a dimens??o ??tica da atividade de agentes p??blicos de um Estado democr??tico, englobando quest??es referentes aos direitos humanos, direitos e deveres do Estado e da sociedade, o papel da transpar??ncia, do controle social e da responsabiliza????o por resultados (accountability) etc. Pode ser aplicado em cursos sobre ??tica e servi??o p??blico, pol??ticas p??blicas de g??nero e ra??a e atendimento ao cidad??o

Eco universidade: plano ambiental para uma universidade socioambientalmente correta

Com a transforma????o da Escola Superior de Agricultura de Lavras na Universidade Federal de Lavras (Ufla), a estrutura existente n??o estava preparada para suportar o crescimento. O saneamento era feito por meio de sumidouros, os res??duos de laborat??rios (qu??micos e biol??gicos) eram descartados de forma inadequada prejudicando o meio ambiente, havia ocorr??ncia de inc??ndios na ??rea do campus. A Administra????o da Ufla estruturou uma s??rie de a????es para solucionar esses problemas. Assim surgiu o Plano Ambiental da Ufla, que aborda conceitos de sustentabilidade e compreendeu a????es que envolvem praticamente todas as quest??es ambientais (implanta????o de Programa de Gerenciamento de Res??duos Qu??micos, tratamento dos res??duos s??lidos, saneamento b??sico, esta????o de tratamento de esgoto, constru????es ecologicamente corretas, prote????o de nascentes e matas ciliares, preven????o e controle de inc??ndios, preven????o de endemias e gest??o de energia). Hoje a Ufla ?? uma universidade sustent??vel

Plano popular de desenvolvimento sustent??vel a jusante da UHE Tucuru??

O setor el??trico brasileiro, historicamente, possui modelo autorit??rio de planejamento e implanta????o dos empreendimentos. Os grandes projetos desenvolvimentistas s??o oriundos do per??odo dos governos militares. Popula????es atingidas por impactos de barragens sempre reivindicaram pol??ticas articuladas de compensa????o por parte das empresas e do governo. A partir de 2003, a Eletronorte adotou o planejamento participativo para elabora????o de um plano de desenvolvimento regional ??? PPDJUS ??? como compensa????o pelos impactos da UHE Tucuru?? nos cinco munic??pios a Jusante, iniciativa sem precedentes no Brasil. O PPDJUS hoje articula as pol??ticas de planejamento e gest??o do territ??rio e de recursos h??dricos do Minist??rio do Desenvolvimento Agr??rio (MDA), do Minist??rio do Meio Ambiente (MMA), do Minist??rio da Integra????o Nacional (MIN), da Secretaria Especial de Abastecimento e Pre??os (Seap), da Ag??ncia de Desenvolvimento da Amaz??nia (ADA) e da Eletronorte, entre outros ??rg??os. Possui conselhos gestores regional e municipais e c??maras t??cnicas, avan??ado modelo de gest??o participativa

Reflex??es para Ibero-Am??rica: avalia????o de programas sociais

A ENAP Escola Nacional de Administra????o P??blica, com o apoio do Minist??rio do Planejamento, Or??amento e Gest??o, da Ag??ncia Brasileira de Coopera????o do Minist??rio das Rela????es Exteriores, do Centro Latino-americano de Administra????o para o Desenvolvimento (Clad) e da Caixa Econ??mica Federal, construiu uma s??rie de atividades de aprendizagem direcionadas a dirigentes e t??cnicos que atuam nas ??reas de planejamento estrat??gico e avalia????o de programas. Esses eventos de aprendizagem permitiram uma intensa troca de experi??ncias entre os 60 dirigentes e t??cnicos representantes de organismos governamentais de 16 pa??ses ibero-americanos ??? Argentina, Bol??via, Brasil, Chile, Col??mbia, Costa Rica, El Salvador, Guatemala, M??xico, Panam??, Paraguai, Peru, Portugal, Rep??blica Dominicana, Uruguai e Venezuela. A consolida????o desse per??odo de rica troca de experi??ncias est?? retratada nesta publica????o, que mostra o avan??o na constru????o de um conhecimento coletivo referente a desafios contempor??neos de nossos governos nas ??reas de planejamento e de avalia????o de programas sociais

Colet??nea planejamento e or??amento governamental: volume 1: o plano como aposta

O texto sintetiza os principais fundamentos do m??todo PES. Esse m??todo contrap??e-se ao planejamento tradicional e prop??e uma abordagem tecnopol??tica do planejamento.

Instrumentos de monitoramento e avalia????o do plano Brasil sem mis??ria e de programas do MDS

Apresenta????o do tema Instrumentos de Monitoramento e Pesquisas de Avalia????o do Plano Brasil sem Mis??ria e de Programas do MDS

Plano integrado de capacita????o dos servidores t??cnico-administrativos

A equipe de Recursos Humanos (PROREH) da Universidade Federal de Uberl??ndia constatou a necessidade de superar a vis??o reducionista da pol??tica de recursos humanos vigente ??? centrada em programas de ???treinamento??? restritos e segmentados ??? e de buscar formas de se adequar ??s exig??ncias de um mundo em mudan??a. A solu????o foi encontrada no estabelecimento de uma pol??tica de capacita????o e desenvolvimento baseada na vis??o do servidor p??blico como cidad??o, inserido na busca dos objetivos da universidade e da sociedade, a partir de uma forma????o global. Como resultado s??o observadas significativas melhorias de desempenho do quadro de pessoal, al??m de manifesta????es positivas das chefias e dos usu??rios dos servi??os

Programa de forma????o educacional b??sica

Desde a d??cada de 1970, tinha sido criado um programa de alfabetiza????o destinado a atingir seus servidores e prestadores de servi??os. Todavia, n??o eram obtidos resultados ??timos. No final dos anos 80, devido a algumas reformas organizacionais internas, esse programa foi alocado na Coordena????o Geral de Recursos Humanos, que passou a desenvolv??-lo como parte das a????es de valoriza????o do servidor. No in??cio da d??cada de 90, o programa foi inclu??do no Plano Decenal de Educa????o para Todos, 1993-2003 e assumiu a denomina????o de Programa de Forma????o Educacional B??sica (PEB).O Programa atingiu a alfabetiza????o de dezenas de alunos. Os que se alfabetizaram inicialmente (Fase I) apresentaram demanda de amplia????o do programa, com a cria????o das Fases II e III subseq??entes. Hoje existem solicita????es de cria????o de uma Fase IV, destinada ao ensino de 2o grau. Constataram-se mudan??as comportamentais positivas, como a eleva????o do n??vel de consci??ncia de direitos e deveres, maior satisfa????o no trabalho, envolvimento das chefias e aumento de interessados no programa, tanto de alunos como de professores