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.

Adjustment of homeless adolescents to a crisis shelter: Application of a stress and coping model

The present study examined the utility of a stress and coping model of adaptation to a homeless shelter among homeless adolescents. Seventy-eight homeless adolescents were interviewed and completed self-administered scales at Time 1 (day of shelter entry) and Time 2 (day of discharge). The mean duration of stay at the shelter was 7.23 days (SD = 7.01). Predictors included appraisal (threat and self-efficacy), coping resources, and coping strategies (productive, nonproductive, and reference to others coping). Adjustment outcomes were Time I measures of global distress, physical health, clinician-and youthworker- rated social adjustment, and externalizing behavior and Time 2 youthworker-rated social adjustment and goal achievement. Results of hierarchical regression analyses indicated that after controlling for the effects of relevant background variables (number of other shelters visited, sexual, emotional, and physical abuse), measures of coping resources, appraisal, and coping strategies evidenced distinct relations with measures of adjustment in ways consistent with the model's predictions with few exceptions. In cross-sectional analyses better Time I adjustment was related to reports of higher levels of coping resources, self-efficacy beliefs, and productive coping strategies, and reports of lower levels of threat appraisal and nonproductive coping strategies. Prospective analyses showed a link between reports of higher levels of reference to others coping strategies and greater goal achievement and, unexpectedly, an association between lower self-efficacy beliefs and better Time 2 youthworker-rated social adjustment. Hence, whereas prospective analyses provide only limited support for the use of a stress and coping model in explaining the adjustment of homeless adolescents to a crisis shelter, cross-sectional findings provide stronger support.

Conservation of the cardiostimulant effects of (-)-Norepinephrine across Ser49Gly and Gly389Arg Beta1-adrenergic receptor polymorphisms in human right atrium in vitro

OBJECTIVES The goal of this study was to determine whether the cardiostimulant effects of the endogenous beta(1)-adrenergic receptor (AR) agonist, (-)-norepinephrine are modified by polymorphic (Serine49Glycine [Ser49Gly], Glycine389Arginine [Gly389Arg]) variants of beta(1)-ARs in the nonfailing adult human heart. BACKGROUND Human heart beta(1)-ARs perform a crucial role in mediating the cardiostimulant effects of (-)-norepinephrine. An understanding of the significance of Ser49Gly and Gly389Arg polymorphisms in the human heart is beginning to emerge, but not as yet in adult patients who have coronary artery disease (CAD). METHODS The potency and maximal effects of (-)-norepinephrine at beta(1)-ARs (in the presence of beta(2)-AR blockade with 50 nM ICI 118,551 [erythro-DL-1(7-methylindan-4-yloxy)-3-isopropylamino-butan-2-ol]) for changes in contractile force and shortening of contractile cycle duration were determined in human right atrium in vitro from 87 patients undergoing coronary artery bypass grafting who were taking beta-blockers before surgery. A smaller sample of patients (n = 20) not taking beta-blockers was also investigated. Genotyping for two beta(1)-AR polymorphisms (Ser49Gly and Gly389Arg) was determined from a sample of blood taken at the time of surgery. RESULTS (-)-Norepinephrine caused concentration-dependent increases in contractile force and reductions in time to reach peak force and time to reach 50% relaxation. There were no differences in the potency or maximal effects of (-)-norepinephrine in the right atrium from patients with different Ser49Gly and Gly389Arg polymorphisms. CONCLUSIONS The cardiostimulant effects of (-)-norepinephrine at beta(1)-ARs were conserved across Ser49Gly and Gly389Arg polymorphisms in the right atrium of nonfailing hearts from patients with CAD managed with or without beta-blockers. (C) 2002 by the American College of Cardiology Foundation.

Conceptions of health and illness held by Australian Aboriginal, Torres Strait Islander and Papua New Guinea Health Science students

Health is considered to be a fundamental human right and developing a better understanding of health is assumed to be a global social goal (Bloom, 1987). Yet many third-world countries and some subpopulations within developed countries do not enjoy a healthy existence. The research reported in this paper examined the conceptions of health and conceptions of illness for a group of Aboriginal, Torres Strait Islander, and Papua New Guinea university students studying health science courses. Results found three conceptions of health and three conceptions of illness that indicated these students held a mix of traditional cultural and Western beliefs. These findings may contribute to overcoming the dissonance between traditional and Western beliefs about health and the development of health care courses that are more specific to how these students understand health. This may also serve to improve the educational status of Aboriginal and Torres Strait Islander people and potentially improve the health status within these communities.

Protection of mice from group A streptococcal infection by intranasal immunisation with a peptide vaccine that contains a conserved M protein B cell epitope and lacks a T cell autoepitope

Infection with group A streptococci (GAS) can lead to rheumatic fever (RF) and rheumatic heart disease (RHD) which are a major health concern particularly in indigenous populations worldwide, and especially in Australian Aboriginals. A primary route of GAS infection is via the upper respiratory tract, and therefore, a major goal of research is the development of a mucosal-based GAS vaccine, The majority of the research to date has focused on the GAS M protein since immunity to GAS is mediated by M protein type-specific opsonic antibodies. There are two major impediments to the development of a vaccine-the variability in M proteins and the potential for the induction of an autoimmune response. To develop a safe and broad-based vaccine, we have therefore focused on the GAS M protein conserved C-region, and have identified peptides, J8 and the closely related J8 peptide (J14), which may be important in protective immunity to GAS infection. Using a mucosal animal model system, our data have shown a high degree of throat GAS colonisation in B10.BR mice 24 h following intranasal immunisation with the mucosal adjuvant, cholera toxin B subunit (CTB), and/or diptheria toxoid (dT) carrier, or PBS alone, and challenge with the M1 GAS strain. However, GAS colonisation of the throat was significantly reduced following intranasal immunisation of mice with the vaccine candidate J8 conjugated to dT or J14-dT when administered with CTB. Moreover, J8-dT/CTB and J14-dT/CTB-immunised mice had a significantly higher survival when compared to CTB and PBS-immunised control mice. These data indicate that immunity to GAS infection can be evoked by intranasal immunisation with a GAS M protein C-region peptide vaccine that contains a protective B cell epitope and lacks a T cell autoepitope. (C) 2002 Published by Elsevier Science Ltd.

Using siting algorithms in the design of marine reserve networks

Using benthic habitat data from the Florida Keys (USA), we demonstrate how siting algorithms can help identify potential networks of marine reserves that comprehensively represent target habitat types. We applied a flexible optimization tool-simulated annealing-to represent a fixed proportion of different marine habitat types within a geographic area. We investigated the relative influence of spatial information, planning-unit size, detail of habitat classification, and magnitude of the overall conservation goal on the resulting network scenarios. With this method, we were able to identify many adequate reserve systems that met the conservation goals, e.g., representing at least 20% of each conservation target (i.e., habitat type) while fulfilling the overall aim of minimizing the system area and perimeter. One of the most useful types of information provided by this siting algorithm comes from an irreplaceability analysis, which is a count of the number of, times unique planning units were included in reserve system scenarios. This analysis indicated that many different combinations of sites produced networks that met the conservation goals. While individual 1-km(2) areas were fairly interchangeable, the irreplaceability analysis highlighted larger areas within the planning region that were chosen consistently to meet the goals incorporated into the algorithm. Additionally, we found that reserve systems designed with a high degree of spatial clustering tended to have considerably less perimeter and larger overall areas in reserve-a configuration that may be preferable particularly for sociopolitical reasons. This exercise illustrates the value of using the simulated annealing algorithm to help site marine reserves: the approach makes efficient use of;available resources, can be used interactively by conservation decision makers, and offers biologically suitable alternative networks from which an effective system of marine reserves can be crafted.

Ecological criteria for evaluating candidate sites for marine reserves

Several schemes have been developed to help select the locations of marine reserves. All of them combine social, economic, and biological criteria, and few offer any guidance as to how to prioritize among the criteria identified. This can imply that the relative weights given to different criteria are unimportant. Where two sites are of equal value ecologically; then socioeconomic criteria should dominate the choice of which should be protected. However, in many cases, socioeconomic criteria are given equal or greater weight than ecological considerations in the choice of sites. This can lead to selection of reserves with little biological value that fail to meet many of the desired objectives. To avoid such a possibility, we develop a series of criteria that allow preliminary evaluation of candidate sites according to their relative biological values in advance of the application of socioeconomic criteria. We include criteria that,. while not strictly biological, have a strong influence on the species present or ecological processes. Out scheme enables sites to be assessed according to their biodiversity, the processes which underpin that diversity, and the processes that support fisheries and provide a spectrum of other services important to people. Criteria that capture biodiversity values include biogeographic representation, habitat representation and heterogeneity, and presence of species or populations of special interest (e.g., threatened species). Criteria that capture sustainability of biodiversity and fishery values include the size of reserves necessary to protect viable habitats, presence of exploitable species, vulnerable life stages, connectivity among reserves, links among ecosystems, and provision of ecosystem services to people. Criteria measuring human and natural threats enable candidate sites to be eliminated from consideration if risks are too great, but also help prioritize among sites where threats can be mitigated by protection. While our criteria can be applied to the design of reserve networks, they also enable choice of single reserves to be made in the context of the attributes of existing protected areas. The overall goal of our scheme is to promote the development of reserve networks that will maintain biodiversity and ecosystem functioning at large scales. The values of eco-system goods and services for people ultimately depend on meeting this objective.

Kangaroos and feral goats as economic resources for graziers: Some views from south-west Queensland

Fifteen years ago it was proposed that the conversion of kangaroos from a pest to an economically valuable resource would allow graziers to reduce the numbers of domestic stock and thereby lower total grazing pressure. Since then, little progress towards this goal has been achieved. This is believed to be due mainly to the low prices obtained for kangaroo products. A survey of graziers in south-west Queensland was carried out to discover their opinions on kangaroos as a potential economic resource. Questions on the harvesting of feral goats were also included in the survey because of the contrast this industry provides to kangaroo harvesting in terms of grazier involvement. The results of the survey are discussed in relation to resource ownership rights; kangaroo product prices and marketing; and competition within the kangaroo harvesting industry. They show that while low kangaroo product prices do act as a disincentive to graziers, other administrative, legal and institutional factors are also important impediments to their entry to the industry. It is concluded that until the focus of attention widens to include consideration of these as well as just market factors, little progress will be made towards integrating graziers into the kangaroo harvesting industry.

Visual Biology of Hawaiian Coral Reef Fishes. III. Environmental Light and an Integrated Approach to the Ecology of Reef Fish Vision

In the previous two papers in this three-part series, we have examined visual pigments, ocular media transmission, and colors of the coral reef fish of Hawaii. This paper first details aspects of the light field and background colors at the microhabitat level on Hawaiian reefs and does so from the perspective and scale of fish living on the reef. Second, information from all three papers is combined in an attempt to examine trends in the visual ecology of reef inhabitants. Our goal is to begin to see fish the way they appear to other fish. Observations resulting from the combination of results in all three papers include the following. Yellow and blue colors on their own are strikingly well matched to backgrounds on the reef such as coral and bodies of horizontally viewed water. These colors, therefore, depending on context, may be important in camouflage as well as conspicuousness. The spectral characteristics of fish colors are correlated to the known spectral sensitivities in reef fish single cones and are tuned for maximum signal reliability when viewed against known backgrounds. The optimal positions of spectral sensitivity in a modeled dichromatic visual system are generally close to the sensitivities known for reef fish. Models also predict that both UV-sensitive and red-sensitive cone types are advantageous for a variety of tasks. UV-sensitive cones are known in some reef fish, red-sensitive cones have yet to be found. Labroid colors, which appear green or blue to us, may he matched to the far-red component of chlorophyll reflectance for camouflage. Red cave/hole dwelling reef fish are relatively poorly matched to the background they are often viewed against but this may be visually irrelevant. The model predicts that the task of distinguishing green algae from coral is optimized with a relatively long wavelength visual pigment pair. Herbivorous grazers whose visual pigments are known possess the longest sensitivities so far found. Labroid complex colors are highly contrasting complementary colors close up but combine, because of the spatial addition, which results from low visual resolution, at distance, to match background water colors remarkably well. Therefore, they are effective for simultaneous communication and camouflage.

Chapter 16: Prophylactic Human Papillomavirus Vaccines

Candidate prophylactic vaccines based on papillomavirus L1 virus-like particles (VLPs) are currently in human clinical trials. The main long-term goal of the vaccine is to reduce the incidence of cervical cancer and its precursors. In animal papillomavirus models, systemic immunization with L1 VLPs can induce high titers of neutralizing antibodies that confer protection against high-dose experimental papillomavirus challenge. In humans, systemic vaccination with L1 VLPs has been well tolerated and induced high serum antibody titers (at least 40 times higher than titers seen following natural infection). A recent proof of principle HPV16 L1 VLP efficacy trial has shown excellent protection against persistent HPV16 infection and associated cytological abnormalities. Large scale efficacy trials of L1 VLPs from HPV16 and 18 (the HPV types found most frequently in cervical cancer), with or without HPV6 and 11 (the HPV types responsible for most genital warts), are planned. If the results of these large trials support the encouraging results of the early trials, they should lead to a commercial prophylactic HPV vaccine. Implementation issues may include how to make the vaccine available in the developing world, where the majority of cervical cancer cases occur, the appropriate age of vaccination, and the role of male vaccination. Because a VLP vaccine is likely to provide type-specific protection, increasing the number of cancer-associated HPV types in the vaccine is a likely approach to broadening the protection to additional types. There will probably also be efforts to develop alternative vaccine formulations better suited to implementation in developing countries as well as attempts to develop vaccines with a therapeutic activity against established HPV infection because a combined prophylactic/therapeutic vaccine may be expected to have an even greater impact than a purely prophylactic vaccine on HPV induced disease.

Aboriginal and Torres Strait Islander health research and the conduct of longitudinal studies: issues for debate

The National Health and Medical Research Council, Research Agenda Working Group (RAWG), and the literature on Indigenous health have identified the need to fill gaps in descriptive data on Aboriginal and Torres Strait Islander health and noted both the lack of research with urban populations and the need for longitudinal studies. This paper presents some of the broad ethical and methodological challenges associated with longitudinal research in Indigenous health and focuses particularly on national studies and studies in urban areas. Our goal is to advance debate in the public health arena about the application of ethical guidelines and the conduct of longitudinal studies in Aboriginal and Torres Strait Islander communities. We encourage others to offer their experiences in this field.

Optical experiments beyond the quantum limit: Squeezing entanglement and teleportation

Results of experiments recently performed are reported, in which two optical parametric amplifiers were set up to generate two independently quadrature squeezed continuous wave laser beams. The transformation of quadrature squeezed states into polarization squeezed states and into states with spatial quantum correlations is demonstrated. By utilizing two squeezed laser beams, a polarization squeezed state exhibiting three simultaneously squeezed Stokes operator variances was generated. Continuous variable polarization entanglement was generated and the Einstein-Podolsky-Rosen paradox was observed. A pair of Stokes operators satisfied both the inseparability criterion and the conditional variance criterion. Values of 0.49 and 0.77, respectively, were observed, with entanglement requiring values below unity. The inseparability measure of the observed quadrature entanglement was 0.44. This value is sufficient for a demonstration of quantum teleportation, which is the next experimental goal of the authors.

Implementa????o e reformula????o de pol??ticas p??blicas: o caso do programa Cultura Viva

O presente artigo tem por objetivo demonstrar como um processo de implementa????o se modifica e ?? adaptado ao longo do tempo, podendo conduzir a um redesenho da pol??tica p??blica. Utiliza-se como estudo de caso o programa Cultura Viva. A pesquisa em que se baseia o artigo foi realizada por meio de entrevistas semiestruturadas com gestores do programa Cultura Viva e gestores de pontos de cultura, bem como por meio de um survey com os pontos de cultura do Estado de S??o Paulo e seus Munic??pios. Os resultados revelam que a implementa????o do programa alterou os crit??rios estabelecidos no processo de formula????o e incluiu novos instrumentos de gest??o, redesenhando assim a pol??tica p??blica

Vantagens e desvantagens do preg??o na gest??o de compras no setor p??blico: o caso da Funasa/PB

O presente artigo ?? parte de um estudo explorat??rio cujo objetivo foi analisar as vantagens e desvantagens na aquisi????o de bens e servi??os por meio das modalidades de licita????o preg??o presencial e eletr??nico no servi??o p??blico. Para isso, foi realizado um estudo de caso na Funda????o Nacional de Sa??de da Para??ba. A parte da pesquisa divulgada neste artigo, al??m de uma s??ntese sobre o referencial te??rico, apresenta as caracter??sticas investigadas da modalidade de licita????o preg??o, comparando suas formas presencial e eletr??nica na institui????o estudada. Os resultados apontam aspectos relevantes quanto ??s quest??es: vantagens e desvantagens do preg??o eletr??nico em rela????o ao preg??o presencial e vice-versa; recursos para atendimento de despesas; planejamento anual da FUNASA/PB; economia de pre??o; economia e repasse de recursos para outras necessidades; prazos de fornecimento; treinamento e capacita????o de pessoal. Conclui a exposi????o destacando alguns pontos fortes e fracos do processo, sugerindo medidas a serem avaliadas no que tange a capacita????o de pessoal e planejamento anual na FUNASA/PB.

Conselhos municipais de pol??ticas p??blicas: uma an??lise explorat??ria

Os Conselhos Gestores s??o f??runs h??bridos onde se viabiliza a participa????o popular no desenho das pol??ticas p??blicas, nas negocia????es de interesses da coletividade, na elabora????o de programas e projetos sociais e na fiscaliza????o das a????es governamentais. O significado e a dimens??o dos Conselhos para a democracia participativa s??o enormes, justificando a import??ncia da realiza????o deste estudo, que se baseou na an??lise explorat??ria de 12 Conselhos locais do munic??pio mineiro de Vi??osa, com o objetivo de avaliar o desempenho dos mesmos. A avalia????o do desempenho levou em considera????o os mesmos par??metros dimensionais, tanto para a coleta de dados prim??rios quanto para a coleta secund??ria. A coleta de dados prim??rios foi realizada a partir da aplica????o de question??rios semi-estruturados aos presidentes dos Conselhos em an??lise. Utilizou-se o programa SPSS, vers??o 15.0, para an??lise estat??stica das quest??es objetivas dos question??rios e a t??cnica de an??lise de conte??do para exame das quest??es abertas. Ap??s essa etapa, realizou-se um workshop com os presidentes dos Conselhos, visando ?? obten????o de novos dados e informa????es. Os Conselhos municipais analisados apresentaram resultados pouco discrepantes entre a teoria aplicada ao tema e a sua efetiva pr??tica no campo p??blico. Contudo, a pesquisa revelou que o grau de desenvolvimento institucional dos Conselhos n??o ?? homog??neo, ou seja, h?? alguns melhores gestores que outros. A escolha independente e democr??tica dos membros n??o-governamentais dos Conselhos em Vi??osa revelou o seu grau de independ??ncia do Poder Executivo. Por??m, a capacita????o de conselheiros precisa ser implementada para evitar uma desfigura????o dos Conselhos. No que tange ?? governan??a, destacouse em Vi??osa o seu grau e formas de aplicabilidade observadas na gest??o de v??rias pol??ticas. Da mesma forma, foi observado um n??vel de governabilidade que confere legitimidade ??s pol??ticas implementadas. Por fim, verificou-se que a efetividade da accountability societal tem sido instrumentalizada pelos Conselhos no ??mbito das diversas pol??ticas setoriais. J?? os dados secund??rios da an??lise foram obtidos por meio das s??nteses dos relat??rios de fiscaliza????o de munic??pios do Estado de Minas Gerais, realizadas pela Controladoria Geral da Uni??o. Foram identificados todos os registros pertinentes aos Conselhos Gestores Municipais do Estado de Minas Gerais, no per??odo de 2003 a 2006. Verificou-se que os maiores problemas se referem aos Conselhos Municipais de Sa??de e Assist??ncia Social. Entre as in??meras falhas apontadas, destacam-se problemas graves na constitui????o e operacionaliza????o dos Conselhos, resultando na inoper??ncia desses mecanismos nos munic??pios fiscalizados. A avalia????o do desempenho dos Conselhos locais de Vi??osa possibilitou concluir que esses mecanismos apresentaram um desempenho bem satisfat??rio na gest??o e controle das pol??ticas p??blicas no munic??pio.