A new green fluorescent protein-based bacterial biosensor for analysing phenanthrene fluxes.

The polycyclic aromatic hydrocarbon (PAH)-degrading strain Burkholderia sp. RP007 served as host strain for the design of a bacterial biosensor for the detection of phenanthrene. RP007 was transformed with a reporter plasmid containing a transcriptional fusion between the phnS putative promoter/operator region and the gene encoding the enhanced green fluorescent protein (GFP). The resulting bacterial biosensor--Burkholderia sp. strain RP037--produced significant amounts of GFP after batch incubation in the presence of phenanthrene crystals. Co-incubation with acetate did not disturb the phenanthrene-specific response but resulted in a homogenously responding population of cells. Active metabolism was required for induction with phenanthrene. The magnitude of GFP induction was influenced by physical parameters affecting the phenanthrene flux to the cells, such as the contact surface area between solid phenanthrene and the aqueous phase, addition of surfactant, and slow phenanthrene release from Model Polymer Release System beads or from a water-immiscible oil. These results strongly suggest that the bacterial biosensor can sense different phenanthrene fluxes while maintaining phenanthrene metabolism, thus acting as a genuine sensor for phenanthrene bioavailability. A relationship between GFP production and phenanthrene mass transfer is proposed.

Hospital outbreak of methicillin-resistant Staphylococcus aureus caused by a new, possibly hyperepidemic variant from a previously sporadic strain.

Objective: To describe an ongoing outbreak that tripled the annual detection of methicillin-resistant Staphylococcus aureus (MRSA) carriage in a tertiary care hospital. Methods: Active surveillance of MRSA is performed since 20 years in our hospital. Our protocol includes screening of patients transferred from high-incidence health-care institutions or countries, roommates of new MRSA cases, and wards where _2 patients acquired MRSA during the same week. Contact precautions are used for known carriers. PFGE was used for molecular typing until 2004, and was then replaced by Double-Locus Sequence Typing (DLST). Results: A median yearly incidence of 173 new carriers of MRSA was observed from 2002 to 2007. Since September 2008, an increasing number of new cases were observed, mainly as successive clusters limited to distinct wards, reaching a total of 398 until October 2009. The yearly incidence of new cases rose to 275 in 2008 and 613 in 2009. 60% of the cases were due to one strain: DLST 4−4, ST 228, SCCmecI. The incidence of new cases due to the previously predominant strains remained unchanged. The epidemic strain corresponded to a new variant of a clone responsible for a previous outbreak in 2001, and only sporadically isolated (mean of 20 cases/year) since then. A case- control study documented a significant association between acquisition of the epidemic strain and a stay in intensive and intermediary care units, a highest number of internal transfers, but did not identify a point source of transmission. Infection control practices and antibiotic policy had remained unchanged for several years. Compliance with handhygiene as monitored yearly was on the rise. Screening of 313 healthcare workers only found one carrier of the epidemic strain lately in the outbreak. Additional infection control measures were enforced, including screening at ICU admission and discharge with PCR-based rapid test, routine screening for all patients leaving epidemic wards, introduction of PCR-based rapid test for contact tracing, additional working forces for environmental disinfection, and hospital-wide education of healthcare workers. However, the outbreak was still ongoing after 5 months. Conclusions: Factors linked to the dissemination of this new variant in our institution remain undetermined. This unresolved outbreak suggests that this new variant acquired hyperepidemic properties, which calls for further investigations.

The clc element of Pseudomonas sp. strain B13, a genomic island with various catabolic properties.

Pseudomonas sp. strain B13 is a bacterium known to degrade chloroaromatic compounds. The properties to use 3- and 4-chlorocatechol are determined by a self-transferable DNA element, the clc element, which normally resides at two locations in the cell's chromosome. Here we report the complete nucleotide sequence of the clc element, demonstrating the unique catabolic properties while showing its relatedness to genomic islands and integrative and conjugative elements rather than to other known catabolic plasmids. As far as catabolic functions, the clc element harbored, in addition to the genes for chlorocatechol degradation, a complete functional operon for 2-aminophenol degradation and genes for a putative aromatic compound transport protein and for a multicomponent aromatic ring dioxygenase similar to anthranilate hydroxylase. The genes for catabolic functions were inducible under various conditions, suggesting a network of catabolic pathway induction. For about half of the open reading frames (ORFs) on the clc element, no clear functional prediction could be given, although some indications were found for functions that were similar to plasmid conjugation. The region in which these ORFs were situated displayed a high overall conservation of nucleotide sequence and gene order to genomic regions in other recently completed bacterial genomes or to other genomic islands. Most notably, except for two discrete regions, the clc element was almost 100% identical over the whole length to a chromosomal region in Burkholderia xenovorans LB400. This indicates the dynamic evolution of this type of element and the continued transition between elements with a more pathogenic character and those with catabolic properties.

Comprehensive approach for the detection of antifungal compounds using a susceptible strain of Candida albicans and confirmation of in vivo activity with the Galleria mellonella model.

An efficient screening strategy for the identification of potentially interesting low-abundance antifungal natural products in crude extracts that combines both a sensitive bioautography assay and high performance liquid chromatography (HPLC) microfractionation was developed. This method relies on high performance thin layer chromatography (HPTLC) bioautography with a hypersusceptible engineered strain of Candida albicans (DSY2621) for bioactivity detection, followed by the evaluation of wild type strains in standard microdilution antifungal assays. Active extracts were microfractionated by HPLC in 96-well plates, and the fractions were subsequently submitted to the bioassay. This procedure enabled precise localisation of the antifungal compounds directly in the HPLC chromatograms of the crude extracts. HPLC-PDA-mass spectrometry (MS) data obtained in parallel to the HPLC antifungal profiles provided a first chemical screening about the bioactive constituents. Transposition of the HPLC analytical conditions to medium-pressure liquid chromatography (MPLC) allowed the efficient isolation of the active constituents in mg amounts for structure confirmation and more extensive characterisation of their biological activities. The antifungal properties of the isolated natural products were evaluated by their minimum inhibitory concentration (MIC) in a dilution assay against both wild type and engineered strains of C. albicans. The biological activity of the most promising agents was further evaluated in vitro by electron microscopy and in vivo in a Galleria mellonella model of C. albicans infection. The overall procedure represents a rational and comprehensive means of evaluating antifungal activity from various perspectives for the selection of initial hits that can be explored in more in-depth mode-of-action studies. This strategy is illustrated by the identification and bioactivity evaluation of a series of antifungal compounds from the methanolic extract of a Rubiaceae plant, Morinda tomentosa, which was used as a model in these studies.

Imagens do sensor MODIS associadas a um modelo agronômico para estimar a produtividade de soja

O objetivo deste trabalho foi estimar a produtividade de soja no Rio Grande do Sul, nas safras de 2000/2001 a 2002/2003, por meio de um modelo agronômico implementado em um Sistema de Informação Geográfica (SIG). Duas abordagens foram utilizadas: o modelo agronômico (AGRO), com valores de índice de área foliar (IAF) obtidos da literatura; e o modelo agronômico-espectral (AGROESPEC), com valores de IAF estimados a partir das imagens MODIS (moderate resolution imaging spectroradiometer). As estimativas de produtividade obtidas pelo modelo foram comparadas àquelas fornecidas pelo Instituto Brasileiro de Geografia e Estatística (IBGE), com o uso do teste t para pares de observação. Nas safras 2000/2001 e 2001/2002, não foram observadas diferenças significativas. Para 2002/2003, o modelo subestimou o valor de produtividade em 7,87 e 7,04%, nas abordagens AGRO e AGROESPEC, respectivamente, em comparação à produtividade fornecida pelo IBGE. Ambas as abordagens do modelo permitiram avaliação objetiva e quantitativa do efeito das condições meteorológicas sobre a produtividade de soja. Entretanto, o AGROESPEC forneceu estimativas mais detalhadas, no que se refere à variação espacial da produtividade, em razão do emprego dos valores de IAF estimados a partir das imagens MODIS.

Mapeamento da temperatura da superfície terrestre com uso do sensor AVHRR/NOAA

O objetivo deste trabalho foi avaliar a adequação do uso do sensor AVHRR/NOAA (Advanced Very High Resolution Radiometer/National Oceanic and Atmospheric Administration) para mapeamento da temperatura da superfície terrestre (TST) no Estado do Rio Grande do Sul, por meio da comparação entre três algoritmos clássicos. Foram comparados os métodos de Becker & Li, Sobrino et al. e Kerr et al. para estimativa das TST mínimas, utilizando imagens noturnas e logo após o amanhecer. Os dados de emissividade e TST foram obtidos por meio de combinações matemáticas da radiação detectada nas faixas do visível, infravermelho próximo e termal do sensor AVHRR/NOAA. O sensor AVHRR é adequado para o mapeamento de TST para as condições do tipo de cobertura do solo que predominam no Rio Grande do Sul, visto que a TST estimada pelos três métodos testados foi próxima à temperatura do ar medida nos locais selecionados. O método de Sobrino et al. foi o mais adequado.

Comportamento do NDVI obtido por sensor ótico ativo em cereais

O objetivo deste trabalho foi avaliar, com um sensor ótico ativo, o comportamento do índice de vegetação por diferença normalizada (NDVI - "normalized difference vegetation index"), nas culturas de trigo, triticale, cevada e milho. Cinco experimentos foram conduzidos no Paraná e São Paulo, com variação de classes de solo, doses e fontes de N, e variedades de trigo. As seguintes variáveis foram avaliadas: NDVI, teor de N foliar, matéria seca e produtividade das culturas. Análises de regressões foram realizadas entre as doses de N aplicadas e NDVI, teor de N foliar, matéria seca e produtividade. Análises de correlação entre as variáveis foram realizadas. O trigo, triticale e cevada apresentaram resposta às aplicações de doses crescentes de N, pelo aumento nas leituras do NDVI, no teor de N foliar e na produtividade. Medido pelo sensor ótico ativo utilizado, o NDVI apresenta alto potencial para manejo do N nas culturas do trigo, triticale e cevada, e baixo potencial para a cultura do milho. Há interferência das variedades de trigo nas leituras do sensor ótico ativo.

Influence of biocontrol strain Pseudomonas fluorescens CHA0 and its antibiotic overproducing derivative on the diversity of root colonizing pseudomonads

Non-target effects of biocontrol strains of Pseudomonas on the population of resident pseudomonads should be assessed prior to their large scale application in the environment. The rifampicin resistant bacterium P. fluorescens CHA0-Rif and its antibiotic overproducing derivative CHA0-Rif/pME3424 were introduced into soil microcosms and the population of resident pseudomonads colonizing cucumber roots was investigated after 10 and 52 days. Both CHA0-Rif and CHA0-Rif/pME3424 displaced a part of the resident pseudomonad population after 10 days. To investigate the population structure, utilization of 10 carbon sources and production of two exoenzymes was assessed for 5600 individual pseudomonad isolates and 1700 isolates were subjected to amplified ribosomal DNA restriction analysis of the spacer region (spacer-ARDRA). After 10 days, only the proportion of pseudomonads able to degrade -tryptophan was reduced in treatments inoculated with either biocontrol strain. In parallel the phenotypic diversity was reduced. These effects were only observed 10 days after inoculation, and they were similar for inoculation with CHA0-Rif and CHA0-Rif/pME3424. Changes in the population structure of resident pseudomonads on cucumber roots during plant growth were more pronounced than changes due to the inoculants. The inoculants did not affect the genotypic diversity detected with spacer-ARDRA, but the genotypic fingerprints corresponded only partially to the phenotypic profiles. Overall CHA0-Rif had a small and transient impact on the population of resident pseudomonads and the effect was essentially the same for the genetically engineered derivative CHA0-

Avaliação de fluxos de calor e evapotranspiração pelo modelo SEBAL com uso de dados do sensor ASTER

O objetivo deste trabalho foi avaliar a eficiência da aplicação do modelo SEBAL em estimar os fluxos de energia em superfície e a evapotranspiração diária, numa extensa área de cultivo de arroz irrigado, no município de Paraíso do Sul, RS, tendo como parâmetros dados do sensor ASTER. As variáveis estudadas constituem importantes parâmetros do tempo e do clima em estudos agrometeorológicos e de racionalização no uso da água. As metodologias convencionais de estimativa desses parâmetros são pontuais e geralmente apresentam incertezas, que aumentam quando o interesse é o comportamento espacial desses parâmetros. Aplicou-se o algoritmo "Surface Energy Balance Algorithm for Land" (SEBAL), em uma imagem do sensor "Advanced Spaceborne Thermal Emission and Reflection Radiometer" (ASTER). As estimativas obtidas foram comparadas com medições em campo, realizadas por uma estação micrometeorológica localizada no interior da área de estudo. As estimativas mais precisas foram as de fluxo de calor sensível e de evapotranspiração diária, e a estimativa que apresentou maior erro foi a do fluxo de calor no solo. A metodologia empregada foi capaz de reproduzir os fluxos de energia em superfície de maneira satisfatória para estudos agrometeorológicos e de rendimento de culturas.

Active strain and activation models in cardiac electromechanics

We present a model for mechanical activation of the cardiac tissue depending on the evolution of the transmembrane electrical potential and certain gating/ionic variables that are available in most of electrophysiological descriptions of the cardiac membrane. The basic idea consists in adding to the chosen ionic model one ordinary differential equation for the kinetics of the mechanical activation function. A relevant example illustrates the desired properties of the proposed model, such as delayed muscle contraction and correct magnitude of the muscle fibers' shortening.

Validação de modelo para predição do potencial produtivo de trigo com sensor óptico ativo

O objetivo deste trabalho foi validar um modelo baseado no índice de vegetação por diferença normalizada (NDVI), para a predição da variabilidade espacial da biomassa acumulada na parte aérea e da produtividade de grãos de trigo. Foram realizadas leituras georreferenciadas de NDVI e biomassa, no final do afilhamento, e produtividade. O modelo proposto permitiu relacionar classes de variabilidade espacial do NDVI e da biomassa, com 81% de correspondência entre elas. Entretanto, a relação entre as classes de variabilidade espacial da produtividade e do NDVI foi menor, com 48% de correspondência.

Altered thymic selection by overexpressing cellular FLICE inhibitory protein in T cells causes lupus-like syndrome in a BALB/c but not C57BL/6 strain.

The cellular FLICE inhibitory protein (c-FLIP) is an endogenous inhibitor of the caspase-8 proapoptotic signaling pathway downstream of death receptors. Recent evidence indicates that the long form of c-FLIP (c-FLIP(L)) is required for proliferation and effector T-cell development. However, the role of c-FLIP(L) in triggering autoimmunity has not been carefully analyzed. We now report that c-FLIP(L) transgenic (Tg) mice develop splenomegaly, lymphadenopathy, multiorgan infiltration, high titers of auto-antibodies, and proliferative glomerulonephritis with immune complex deposition in a strain-dependent manner. The development of autoimmunity requires CD4(+) T cells and may result from impaired thymic selection. At the molecular level, c-FLIP(L) overexpression inhibits the zeta chain-associated protein tyrosine kinase of 70 kDa (ZAP-70) activation, thus impairing the signaling pathway derived from ZAP-70 required for thymic selection. Therefore, we have identified c-FLIP(L) as a susceptibility factor under the influence of epistatic modifiers for the development of autoimmunity.

Sensor óptico no auxílio à recomendação de adubação nitrogenada em cana-de-açúcar

O objetivo deste trabalho foi avaliar o potencial de um sensor óptico ativo terrestre como auxiliar na recomendação da aplicação de nitrogênio em taxa variável, na cultura da cana-de-açúcar. Foram instalados experimentos em delineamento de blocos ao acaso, com uso de diferentes doses de N (0, 50, 100, 150 e 200 kg ha-1). A resposta da cana-de-açúcar ao N foi avaliada por diferentes métodos - sensor óptico, clorofilômetro e teor foliar de N -, quando a altura média dos colmos atingiu 0,2, 0,4 e 0,6 m. Observou-se baixa correlação entre o teor foliar de N e a quantidade de clorofila nas folhas mensuradas por clorofilômetro. Portanto, essas características foram insuficientes para avaliar a eficiência do sensor óptico, uma vez que os valores mensurados se elevaram conforme o aumento da dose de N. A estratégia de recomendação com base na resposta da cultura, estimada pelo sensor óptico em faixa de cana-de-açúcar que recebeu a dose adequada de N, mostrou-se mais condizente com a produtividade obtida. O sensor óptico é ferramenta útil para auxiliar na recomendação de N para a cultura da cana-de-açúcar, ao se considerar a variabilidade espacial da sua demanda.

Imagens multipolarizadas do sensor Palsar/Alos na discriminação das fases fenológicas da cana‑de‑açúcar

O objetivo deste trabalho foi avaliar o potencial das imagens multipolarizadas do sensor‑radar Palsar/Alos em diferenciar as fases fenológicas da cana‑de‑açúcar. Valores digitais de quatro imagens do sensor, dos meses de fevereiro, maio, agosto e outubro de 2008, com polarizações HH (emissão e recebimento de onda na polarização horizontal) e HV (emissão de onda na polarização horizontal e recebimento na vertical), foram convertidos para coeficientes de retroespalhamento (σ°), para a análise de dados de cana‑de‑açúcar, cultivadas em talhões na região nordeste do Estado de São Paulo. Foram selecionadas três variedades, em diferentes estágios fenológicos: RB85‑5156, seis talhões; RB86‑7515, dez talhões; e RB92‑5345, dez talhões. As diferenças entre as fases fenológicas foram avaliadas para cada uma das variedades e, também, entre as variedades. A utilização simultânea ou não dos dados do sensor Palsar/Alos, obtidos em duas polarizações, foi capaz de discriminar as diferentes fases de crescimento da cana‑de‑açúcar, com exceção da fase de crescimento dos colmos e a fase de maturação, em que não foi observada diferença significativa.