Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

Produção in vitro de embriões de ovinos: uma visão crítica do método e de seu resultado a campo

A metodologia de produção in vitro de embriões de ovinos implica no desenvolvimento de meios de maturação, fertilização e cultivo que permitam aumentar a taxa de clivagem e desenvolvimento, tanto para o investimento biotecnológico em programas comerciais, quanto para sua utilização em clonagem e transgenia dessa espécie animal. Do ponto de vista da pesquisa, os ovócitos podem ser obtidos pelas técnicas de punção e slicing a partir de ovários oriundos de matadouros, ou através de aspiração folicular por laparoscopia. Como vantagem, este método permite o uso de uma mesma doadora estimulada hormonialmente em intervalos periódicos, mantida sob rigoroso controle sanitário, o que é de vital importância para a produção de biofármacos em programas que utilisem os ovinos como modelo biológico. Por outro lado, em nosso país a demanda pela multiplicação de animais de alto valor genético, seja pela produtividade ou pelo elevado valor comercial dos mesmos, impõe o desenvolvimento, adaptação e otimização das diferentes metodologias desenvolvidas ao longo dos ultimos anos em laboratórios de referência mundiais. Nesse contexto, cresce de importância o perfeito conhecimento da fisiologia dessa espécie e das raças criadas em nosso país, e da problemática da produção in vitro de seus embriões. Respeitando essas premissas, gerar o desenvolvimento de protocolos que permitam não apenas aumentar a população de ovócitos passíveis de maturação in vitro, mas de sua competência ao desenvolvimento ao estágio de blastocisto, ou, alternativamente, sua transferência a receptoras em estágios precoces do desenvolvimento, evitando assim as conhecidas perdas durante o desenvolvimento in vitro, e o baixo percentual de gestações que chegam a termo, com cordeiro saudáveis. Trata-se de um desafio, que já apresenta os primeiros resultados em nosso país, tanto na produção comercial de embriões produzidos in vitro, quanto em programas de clonagem e transgenia.

Environmental conditions and rodent infestation in Campo Limpo district, Sao Paulo municipality, Brazil

Rodents are involved in the transmission to human beings of several diseases, including liptospirosis, which shows high lethality rates in Sao Paulo municipality. Despite this, few studies have assessed the relationship existing between urban environmental conditions and building rodent infestation. With the purpose of clarifying this relationship, an analysis has been conducted in order to quantify the influence of environmental factors upon rodent infestation on a low-income district. Diagnosis of the environmental situation has been performed to evaluate the frequency according to which harborage, food and access sources occur, and a survey on infestation rates in 2175 dwellings in the area studied. The logistic regression analysis showed that among the environmental variables, the one that showed the closest association with rodent infestation was access; followed by harborage, and food. It was concluded that poor socioeconomic and environmental conditions in the area propitiate the occurrence of high rodent infestation rates

Cuidado à saúde de crianças Kaiowá e Guarani: notas de observação de campo

Inicia-se uma discussão sobre as práticas de saúde e cuidado utilizadas pelos Kaiowá e Guarani da Terra Indígena de Caarapó no enfrentamento das doenças diarréicas na infância. Utilizou-se de uma contribuição etnográfica, através do uso da técnica da observação participante e de entrevistas aberta com moradores da comunidade. Em se tratando de uma sociedade que vivencia permanentes transformações de ordem social advindas das relações interétnicas e da sucessiva presença dos serviços de saúde, observa-se que o significado de diarréia infantil, assim como as decisões relativas à prevenção e ao seu tratamento, refletem comportamentos diferenciados e complexos. A diarréia, também conhecida como chiri, é definida através de sinais que, de certa forma, são semelhantes aos biomédicos. No entanto, nem sempre as explicações de causalidade e as formas de tratamento seguem, unicamente, a abordagem biomédica. A variedade de causas da doença diarréica entre as crianças indígenas implica na escolha do processo terapêutico, que pode ser desde a procura por um especialista tradicional, realização de chás e infusões e, ainda, procura pelos serviços de saúde. Tais explicações e formas de tratamento pressupõem a existência de um processo de negociação entre pessoas de uma mesma cultura e de culturas distintas. Neste sentido, os serviços de saúde ao tratar a criança com diarréia, devem considerar não apenas a perspectiva biomédica, mas também dialogar com a percepção e as práticas indígenas na identificação da causalidade, na definição do diagnóstico e do tratamento, sobretudo, pela coexistência destas práticas no contexto local

Sobre as dores e temores do parto: dimensões de uma escuta

OBJETIVOS: desenvolver algumas reflexões sobre os possíveis efeitos benéficos de uma escuta responsiva à verbalização da presença de dor, medos e seus correlatos na cena do parto tomando como base dados empíricos de pesquisa realizada em maternidade situada na cidade de São Paulo, Brasil. MÉTODOS: estudo descritivo, de metodologia qualitativa, referenciado no quadro teórico da Psicologia Social, Psicanálise e Lingüística, utilizando entrevistas semi-estruturadas com 20 parturientes e sete doulas e observação de rotinas da maternidade. RESULTADOS: a análise, apoiada em categorias estabelecidas (subjetividade auto-referida, intersubjetividade, acolhimento, apropriação da experiência) mostrou, entre outros pontos, a importância e a valorização da interlocução qualificada no processo da parturição. Processo esse referido pelas parturientes como experiência de elevado grau de estresse, com vivências de dor, medos e ansiedades, porém mitigados pelo apoio recebido. CONCLUSÕES: a análise permitiu compreender as relações interpessoais como campo de interlocução e acolhimento percebidos pelas mulheres do estudo capazes de produzir efeitos favoráveis sobre as vivências do estresse materno, configurando-se como recurso técnico, qualificado e valioso, oferecido à parturiente

Bioeticistas brasileiros e os princípios da universalidade e da integralidade no SUS

OBJETIVO: Compreender os sentidos que bioeticistas brasileiros atribuem aos princípios da universalidade e da integralidade no sistema público de saúde brasileiro. PROCEDIMENTOS METODOLÓGICOS: Estudo exploratório qualitativo, realizado com 20 professores universitários de bioética atuantes no campo das ciências da saúde, com funções de diretores e ex-diretores da Sociedade Brasileira de Bioética e de diretorias regionais, no período de julho de 2007 a julho de 2008. Foram realizadas entrevistas semi-estruturadas, com perguntas abertas, sendo realizada análise de conteúdo. ANÁLISE DE RESULTADOS: Quanto ao princípio da universalidade de acesso dos cidadãos brasileiros a um sistema público, as manifestações dos entrevistados se posicionaram majoritariamente em prol de sua manutenção. Todavia, quanto ao princípio da integralidade, as divergências foram manifestas, ensejando a maioria em restringi-lo. CONCLUSÕES: Os bioeticistas relatam pluralismo de valores morais e dificuldades em decidir moralmente sobre o que seria um sistema de saúde justo

Antileishmanial Activity of the Hydroalcoholic Extract of Miconia langsdorffii, Isolated Compounds, and Semi-Synthetic Derivatives

The in vitro activity of the crude hydroalcoholic extract of the aerial parts of Miconia langsdorffii Cogn. was evaluated against the promastigote forms of L. amazonensis, the causative agent of cutaneous leishmaniasis in humans. The bioassay-guided fractionation of this extract led to identification of the triterpenes ursolic acid and oleanolic acid as the major compounds in the fraction that displayed the highest activity. Several ursolic acid semi-synthetic derivatives were prepared, to find out whether more active compounds could be obtained. Among these ursolic acid-derived substances, the C-28 methyl ester derivative exhibited the best antileishmanial activity.

Quantitative constraints on the transport properties of hot partonic matter from semi-inclusive single high transverse momentum pion suppression in Au plus Au collisions at root S(NN)=200 GeV

The PHENIX experiment has measured the suppression of semi-inclusive single high-transverse-momentum pi(0)'s in Au+Au collisions at root s(NN) = 200 GeV. The present understanding of this suppression is in terms of energy loss of the parent (fragmenting) parton in a dense color-charge medium. We have performed a quantitative comparison between various parton energy-loss models and our experimental data. The statistical point-to-point uncorrelated as well as correlated systematic uncertainties are taken into account in the comparison. We detail this methodology and the resulting constraint on the model parameters, such as the initial color-charge density dN(g)/dy, the medium transport coefficient <(q) over cap >, or the initial energy-loss parameter epsilon(0). We find that high-transverse-momentum pi(0) suppression in Au+Au collisions has sufficient precision to constrain these model-dependent parameters at the +/- 20-25% (one standard deviation) level. These constraints include only the experimental uncertainties, and further studies are needed to compute the corresponding theoretical uncertainties.

A Semi-Tethered Test for Power Assessment in Running

We analyzed the usefulness of a semi-tethered field running test (STR) and the relationships between indices of anaerobic power, anaerobic capacity and running performance in 9 trained male sprinters (22.2 +/- 2.9 yrs, 176 +/- 1 cm, 68.0 +/- 9.4 kg). STR involved an all out 120 m run attached to an apparatus that enabled power calculation from force and velocity measures. Subjects also carried out a cycloergometer Win-gate Anaerobic Test (WT), an all out 300 m run and had accessed their maximal accumulated oxygen deficit (MAOD) on a treadmill. Peak and mean powers attained in STR (1 720 +/- 221 and 1 391 +/- 201 W) were greater but significantly related (r=0.82; P<0.01) to those in the WT (808 +/- 130 and 603 +/- 87 W). In addition, power measures derived from the STR were stronger related to running performance compared to those from the WT (r=0.81-0.94 vs. 0.68-0.84; P<0.05). Relationships between MAOD and most power indices were only weak to moderate. These results support the usefulness of STR for specific power assessment in field running and suggest that anaerobic power and capacity are not related entities, irrespective of having been evaluated using similar or dissimilar exercise modes.

Analysis of a semi-rigid connection for precast concrete

This paper presents a study of a specific type of beam-to-column connection for precast concrete structures. Furthermore, an analytical model to determine the strength and the stiffness of the connection, based on test results of two prototypes, is proposed. To evaluate the influence of the strength and stiffness of the connection on the behaviour of the structure, the results of numerical simulations of a typical multi-storey building with semi-rigid connections are also presented and compared with the results using pinned and rigid connections. The main conclusions are: (a) the proposed design model can reasonably evaluate the studied connection strength; (b) the evaluation of strength is more accurate than that of stiffness; (c) for a typical structure, it is possible to increase the number of storeys of the structure from two to four with lower horizontal displacement at the top, and only a small increase of the column base bending moment by replacing the pinned connections with semi-rigid ones; and (d) although there is significant uncertainty in the connection stiffness, the results show that the displacements at the top of the structure, and the column base moments present low susceptibility deviations to this parameter.

Magnetic losses evolution of a semi-processed steel during forced aging treatments

An ultra-low carbon steel (30 ppm after decarburization) containing Al and Si was aged for distinct soaking times at 210 degrees C. The core loss increased continuously until around 24 h. After that, only slight changes were verified. It was found that only the hysteresis loss component changed during the aging treatment. By internal friction test and transmission electron microscopy it was seen that carbon precipitation caused the magnetic aging. By scanning electron microscopy it could be concluded that the increase of aging index was attributed to the high number of carbides larger than 0.1 mu m. (C) 2008 Elsevier B. V. All rights reserved.

Ultra-thin films of alternating semi-interpenetrating layers of a conducting polymer with thermosetting phenolic resins for sensor application

In this work a new method for crosslinking ultra-thin films with potential applications in sensor systems is proposed. The films were produced by layer-by-layer (LbL) assembly using a conducting polymer, poly(o-ethoxyaniline) (POEA), alternated with a thermosetting resin, novolac-type phenolformaldehyde (PF), crosslinked by a simple thermal treatment. The PF resin served as both alternating and crosslinking agents. The films were characterized by Fourier transform infrared (FTIR) and ultraviolet-visible (UV-Vis) spectroscopy, thermogravimetry (TG), desorption, doping/dedoping cycling and electrical measurements. The results showed that film architecture and crosslinking degree can be controlled by the conditions used for film deposition (number of bilayers, polymer concentration, pH, and deposition time), and crosslinking time. Moreover, this approach offers several advantages such as fast curing time and low cost, indicating that these films can be used to produce sensors with improved stability.

Repeated disturbances and canopy disturbance regime in a tropical semi-deciduous forest

The canopy disturbance regime and the influence of gap methods on the interpretation of forest structure and dynamics were evaluated in a tropical semi-deciduous forest in south-eastern Brazil. We encountered a gap density of 11.2 gaps ha(-1) and an average size which varied from 121 to 333 m(2) depending on the gap delimitation method considered (minimum gap size was 10 m(2)). Although average size was slightly higher, the median value obtained (78 m(2)) was comparable to other tropical forest sites and the gap size-class distribution found supported the pattern described for such forest sites. Among 297 gap makers, snapping and uprooting were the most common modes of disturbance. The number and basal area of gap makers were good predictors of gap size. Almost 25% of all gaps suffered from repeated disturbance events that brought about larger gap sizes. Such processes, along with delimitation methods, strongly influenced the estimation of turnover rate and therefore the interpretation of forest dynamics. These results demonstrated the importance of further studies on repeated disturbances, which is often neglected in forest studies.

Toward sustainable soil and water resources use in China`s highly erodible semi-arid loess plateau

The water-wind crisscross region of the Loess Plateau in China is comprised of 17.8 million hectares of highly erodible soil under limited annual rainfall. This requires a sustainable water balance for the restoration of dryland ecosystems to reduce and manage soil erosion. In this region, alfalfa has been one of the main legumes grown to minimize soil erosion. However, alfalfa yields were significantly lower in years of reduced rainfall suggesting that high water use and deep rooting alfalfa make it an unsustainable crop due to the long-term decline in soil water storage and productivity. Our objectives in this Study were to evaluate the soil water balance of Loess Plateau soils during vegetative restoration and to evaluate practices that prevent soil desiccation and promote ecosystem restoration and sustainability. Field observations of soil moisture recovery and soil erosion were carried out for five years after alfalfa was replaced with different crops and with bare soil. Soil water content changes in cropland, rangeland, and bare soil were tracked over several years, using a water balance approach. Results indicate that growing forages significantly reduced runoff and sediment transport. A forage-food-crop rotation is a better choice than other cropping systems for achieving sustainable productivity and preventing soil erosion and desiccation. However, economic considerations have prevented its widespread adoption by local farmers. Alternatively, this study recommends consideration of grassland crops or forest ecosystems to provide a sustainable water balance in the Loess Plateau of China. (C) 2009 Elsevier B.V. All rights reserved.