752 research outputs found

    ITS (Intergenic Transcribed Spacer) e Íntron-RFLP para caracterização de Colletotrichum spp. isolado de guaranazeiro.

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    O objetivo do trabalho foi utilizar as regiões ITS e o íntron do gene GS (Glutamine Synthetase) na caracterização de Colletotrichum spp., obtidos a partir de isolamento direto das lesões foliares típicas de antracnose em guaranazeiro

    Structure-function analysis of the curli accessory protein CsgE defines surfaces essential for coordinating amyloid fiber formation

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    Curli amyloid fibers are produced as part of the extracellular biofilm matrix and are composed primarily of the major structural subunit CsgA. The CsgE chaperone facilitates the secretion of CsgA through CsgG by forming a cap at the base of the nonameric CsgG outer membrane pore. We elucidated a series of finely tuned nonpolar and charge-charge interactions that facilitate the oligomerization of CsgE and its ability to transport unfolded CsgA to CsgG for translocation. CsgE oligomerization in vitro is temperature dependent and is disrupted by mutations in the W48 and F79 residues. Using nuclear magnetic resonance (NMR), we identified two regions of CsgE involved in the CsgE-CsgA interaction: a head comprising a positively charged patch centered around R47 and a stem comprising a negatively charged patch containing E31 and E85. Negatively charged residues in the intrinsically disordered N- and C-terminal “tails” were not implicated in this interaction. Head and stem residues were mutated and interrogated using in vivo measurements of curli production and in vitro amyloid polymerization assays. The R47 head residue of CsgE is required for stabilization of CsgA- and CsgE-mediated curli fiber formation. Mutation of the E31 and E85 stem residues to positively charged side chains decreased CsgE-mediated curli fiber formation but increased CsgE-mediated stabilization of CsgA. No single-amino-acid substitutions in the head, stem, or tail regions affected the ability of CsgE to cap the CsgG pore as determined by a bile salt sensitivity assay. These mechanistic insights into the directed assembly of functional amyloids in extracellular biofilms elucidate possible targets for biofilm-associated bacterial infections.Curli represent a class of functional amyloid fibers produced by Escherichia coli and other Gram-negative bacteria that serve as protein scaffolds in the extracellular biofilm matrix. Despite the lack of sequence conservation among different amyloidogenic proteins, the structural and biophysical properties of functional amyloids such as curli closely resemble those of amyloids associated with several common neurodegenerative diseases. These parallels are underscored by the observation that certain proteins and chemicals can prevent amyloid formation by the major curli subunit CsgA and by alpha-synuclein, the amyloid-forming protein found in Lewy bodies during Parkinson’s disease. CsgA subunits are targeted to the CsgG outer membrane pore by CsgE prior to secretion and assembly into fibers. Here, we use biophysical, biochemical, and genetic approaches to elucidate a mechanistic understanding of CsgE function in curli biogenesis

    Desempenho agronômico de grupos de cultivares de alface sistema orgânico no Distrito Federal.

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    Este trabalho teve com objetivo avaliar o desempenho agronômico de cultivares de alface crespa, lisa e americana cultivadas em sistema orgãnico.Trabalho apresentado no 4. Seminário de Agroecologia do Distrito Federal e Entorno, Brasília, DF

    Thermographic inspection of impact damage in carbon fiber-reinforcing polymer matrix laminates

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    Laminados compósitos com matrizes poliméricas, respectivamente termorrígida e termoplástica, fortalecidas com fibras contínuas de carbono foram submetidos a impacto único transversal com diferentes níveis de energia. Os danos impingidos aos materiais estruturais foram avaliados por termografia ativa infravermelha na modalidade transmissão. Em geral, os termogramas do laminado termoplástico apresentaram indicações mais claras e bem definidas dos danos causados por impacto, se comparados aos do compósito termorrígido. O aquecimento convectivo das amostras por fluxo controlado de ar quente se mostrou mais eficaz que o realizado por irradiação, empregando-se lâmpada de filamento. Observou-se também que tempos mais longos de aquecimento favoreceram a visualização dos danos. O posicionamento da face impactada do espécime, relativamente à câmera infravermelha e à fonte de calor, não afetou a qualidade dos termogramas no caso do laminado termorrígido, enquanto que influenciou significativamente os termogramas do compósito termoplástico. Os resultados permitiram concluir que a termografia infravermelha é um método de ensaio não-destrutivo simples, robusto e confiável para a detecção de danos por impacto tão leve quanto 5 J em laminados compósitos poliméricos reforçados com fibras de carbono.Continuous carbon fiber reinforced thermoset and thermoplastic composite laminates were exposed to single transversal impact with different energy levels. The damages impinged to the structural materials were evaluated by active infrared thermography in the transmission mode. In general, the thermoplastic laminate thermograms showed clearer damage indications than those from the thermosetting composite. The convective heating of the samples by controlled hot air flow was more efficient than via irradiation using a filament lamp. It was also observed that longer heating times improved the damage visualization. The positioning of the specimen's impacted face regarding the infrared camera and the heating source did not affect the thermo-imaging of thermosetting specimens, whereas it substantially influenced the thermograms of thermoplastic laminates. The results obtained allow concluding that infrared thermography is a simple, robust and trustworthy methodology for detecting impact damages as slight as 5 J in carbon fiber composite laminates.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)CNPqMaterials Institute of Brazi

    Adaptability and Stability of Alfalfa Cultivars

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    Yield adaptability and stability of thirty five alfalfa genotypes (Medicago sativa L.) were evaluated at Sertãozinho, SP, Brazil, considering different environments (seasons of year) from 1996 to 1999. A randomized complete block design with three replications was used. Scott-Knott’s test was performed to compare genotypes averages. Adaptability and stability were determined by Eberhart and Russell\u27s procedures. Two diverse groups of genotypes with mean dry matter yield were identified: Araucana, F686, MH4 e BR2 were more responsive under favorable environments, but SW 8210, Alto, Monarca SP, Victoria SP, Florida 77, P5888, MH15, BR1, BR3, SW 9210A presented broad adaptability and high predictive stability for diverse environments. It was concluded that several studied genotypes compared to cv. ‘Crioula’, which is traditionally used in Brazil, and these genotypes can be grown in these environments, showing high yielding and broad adaptability associated to phenotypic stability

    Measurement of the cosmic ray spectrum above 4×10184{\times}10^{18} eV using inclined events detected with the Pierre Auger Observatory

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    A measurement of the cosmic-ray spectrum for energies exceeding 4×10184{\times}10^{18} eV is presented, which is based on the analysis of showers with zenith angles greater than 6060^{\circ} detected with the Pierre Auger Observatory between 1 January 2004 and 31 December 2013. The measured spectrum confirms a flux suppression at the highest energies. Above 5.3×10185.3{\times}10^{18} eV, the "ankle", the flux can be described by a power law EγE^{-\gamma} with index γ=2.70±0.02(stat)±0.1(sys)\gamma=2.70 \pm 0.02 \,\text{(stat)} \pm 0.1\,\text{(sys)} followed by a smooth suppression region. For the energy (EsE_\text{s}) at which the spectral flux has fallen to one-half of its extrapolated value in the absence of suppression, we find Es=(5.12±0.25(stat)1.2+1.0(sys))×1019E_\text{s}=(5.12\pm0.25\,\text{(stat)}^{+1.0}_{-1.2}\,\text{(sys)}){\times}10^{19} eV.Comment: Replaced with published version. Added journal reference and DO

    The state of the Martian climate

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    60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

    Proteasome-Dependent Disruption of the E3 Ubiquitin Ligase Anaphase-Promoting Complex by HCMV Protein pUL21a

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    This work is licensed under a Creative Commons Attribution 4.0 International License.The anaphase-promoting complex (APC) is an E3 ubiquitin ligase which controls ubiquitination and degradation of multiple cell cycle regulatory proteins. During infection, human cytomegalovirus (HCMV), a widespread pathogen, not only phosphorylates the APC coactivator Cdh1 via the multifunctional viral kinase pUL97, it also promotes degradation of APC subunits via an unknown mechanism. Using a proteomics approach, we found that a recently identified HCMV protein, pUL21a, interacted with the APC. Importantly, we determined that expression of pUL21a was necessary and sufficient for proteasome-dependent degradation of APC subunits APC4 and APC5. This resulted in APC disruption and required pUL21a binding to the APC. We have identified the proline-arginine amino acid pair at residues 109–110 in pUL21a to be critical for its ability to bind and regulate the APC. A point mutant virus in which proline-arginine were mutated to alanines (PR-AA) grew at wild-type levels. However, a double mutant virus in which the viral ability to regulate the APC was abrogated by both PR-AA point mutation and UL97 deletion was markedly more attenuated compared to the UL97 deletion virus alone. This suggests that these mutations are synthetically lethal, and that HCMV exploits two viral factors to ensure successful disruption of the APC to overcome its restriction on virus infection. This study reveals the HCMV protein pUL21a as a novel APC regulator and uncovers a unique viral mechanism to subvert APC activity
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