993 resultados para spacing between nozzles
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Pós-graduação em Geociências e Meio Ambiente - IGCE
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Pós-graduação em Geociências e Meio Ambiente - IGCE
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Cultured fibroblasts adhere to extracellular substrates by means of cell-matrix adhesions that are assembled in a hierarchical way, thereby gaining in protein complexity and size. Here we asked how restricting the size of cell-matrix adhesions affects cell morphology and behavior. Using a nanostencil technique, culture substrates were patterned with gold squares of a width and spacing between 250 nm and 2 µm. The gold was functionalized with RGD peptide as ligand for cellular integrins, and mouse embryo fibroblasts were plated. Limiting the length of cell-matrix adhesions to 500 nm or less disturbed the maturation of vinculin-positive focal complexes into focal contacts and fibrillar adhesions, as indicated by poor recruitment of ?5-integrin. We found that on sub-micrometer patterns, fibroblasts spread extensively, but did not polarize. Instead, they formed excessive numbers of lamellipodia and a fine actin meshwork without stress fibers. Moreover, these cells showed aberrant fibronectin fibrillogenesis, and their speed of directed migration was reduced significantly compared to fibroblasts on 2 µm square patterns. Interference with RhoA/ROCK signaling eliminated the pattern-dependent differences in cell morphology. Our results indicate that manipulating the maturation of cell-matrix adhesions by nanopatterned surfaces allows to influence morphology, actin dynamics, migration and ECM assembly of adhering fibroblasts.
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We present steady-state absorption and emission spectroscopy and femtosecond broadband photoluminescence up-conversion spectroscopy studies of the electronic relaxation of Os(dmbp)3 (Os1) and Os(bpy)2(dpp) (Os2) in ethanol, where dmbp is 4,4′-dimethyl-2,2′-biypridine, bpy is 2,2′-biypridine, and dpp is 2,3-dipyridyl pyrazine. In both cases, the steady-state phosphorescence is due to the lowest 3MLCT state, whose quantum yield we estimate to be ≤5.0 × 10–3. For Os1, the steady-state phosphorescence lifetime is 25 ns. In both complexes, the photoluminescence excitation spectra map the absorption spectrum, pointing to an excitation wavelength-independent quantum yield. The ultrafast studies revealed a short-lived (≤100 fs) fluorescence, which stems from the lowest singlet metal-to-ligand-charge-transfer (1MLCT) state and decays by intersystem crossing to the manifold of 3MLCT states. In addition, Os1 exhibits a 50 ps lived emission from an intermediate triplet state at an energy 2000 cm–1 above that of the long-lived (25 ns) phosphorescence. In Os2, the 1MLCT–3MLCT intersystem crossing is faster than that in Os1, and no emission from triplet states is observed other than the lowest one. These observations are attributed to a higher density of states or a smaller energy spacing between them compared with Os1. They highlight the importance of the energetics on the rate of intersystem crossing.
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Transcription factors often determine cell fate and tissue development. Chondrogenesis is the developmental process by which cartilages form. Recently, gene targeting studies have shown that two transcription factors, L-Sox5 and Sox6, play essential and redundant roles in chondrogenesis in vivo by converting precartilaginous cell condensations into cartilages. Both are highly similar High-Mobility-Group (HMG)-domain proteins that bind and subsequently bend DNA containing the 7bp HMG site (A/T)(A/T)CAA(A/T)G. They have no transactivation domain, but homo- and hetero-dimerize and preferentially bind DNA containing two HMG sites. They are thought to play an architectural role in transactivation by facilitating long-range DNA and protein interactions. To understand their molecular mechanism of action, we investigated how phasing, orientation, and spacing between HMG sites affect L-Sox5 and Sox6 DNA-binding. We determined that L-Sox5 and Sox6 dimers bind with high affinity to paired HMG sites in DNA rather than a single HMG site. Binding of paired sites is independent of DNA helical phasing, orientation of paired HMG sites and independent of distance up to 255 base pairs between sites. Mutational analysis demonstrated that binding of L-Sox5 and Sox6, independent of orientation of the sites, is critically dependent on the presence of paired HMG sites rather than one HMG site alone. Our data support a unique and novel model whereby L-Sox5 and Sox6 dimerize and bind DNA with pronounced spatial flexibility, possibly by a flexible hinge, and act as architectural transcription factors that bring distant DNA sites and proteins together to form higher order transcriptional complexes that are essential for the activation of their target genes in chondrogenesis. ^
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El presente trabajo tiene por objeto evaluar la uniformidad del riego por goteo en las zonas de regadío de las cuencas de los ríos Mendoza y Tunuyán (zonas Alta y Baja), de la provincia de Mendoza, Argentina. Las evaluaciones -en 17 propiedades- permitieron determinar: coeficientes de uniformidad, salinidad del agua de riego y del suelo en cabeza, medio y pie de la subunidad de riego, y en el bulbo húmedo e interfilar en dos estratos del perfil del suelo (0,10‑0,30 m y 0,30-0,50 m). Se determinó, además, la textura del suelo y su posible relación con los niveles de salinidad de la rizósfera. Se estimó que el 18% de las subunidades de riego evaluadas presentan un coeficiente de uniformidad por debajo del rango recomendable y que en el 94% de las propiedades existen diferencias significativas entre caudales medios registrados entre subunidades y entre sectores de operación de riego. Se encontraron diferencias significativas en la salinidad del extracto de saturación, en la sodicidad y en la concentración del anión cloruro, al comparar el suelo extraído del bulbo de mojado respecto del interfilar. Asimismo, los resultados muestran que no existen diferencias significativas de esas variables en las distintas profundidades de suelo analizadas. El aumento de la salinidad resultó en función del lugar de muestreo (bulbo o interfilar), la calidad del agua de riego y la textura del suelo. La variación de sodicidad, en cambio, dependió del lugar de muestreo (bulbo o interfilar) y del contenido de bicarbonatos en el suelo. Los resultados indican la importancia de realizar evaluaciones rutinarias del comportamiento de los sistemas de riego en términos de uniformidad y salinización inducida.
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In overhead conductor rail lines, aluminium beams are usually mounted with support spacing between 8 and 12 meters, to limit the maximum vertical deflection in the center of the span. This small support spacing limits the use of overhead conductor rail to tunnels, therefore it has been used almost exclusively in metropolitan networks, with operation speeds below 110 km/h. Nevertheless, due to the lower cost of maintenance required for this electrification system, some railway administrations are beginning to install it in some tunnels on long-distance lines, requesting higher operation speeds [1]. Some examples are the Barcelona and Madrid suburban networks (Spain), and recent lines in Turkey and Malaysia. In order to adapt the design of the overhead conductor for higher speeds (V > 160 km/h), particular attention must be paid to the geometry of the conductor rail in critical zones as overlaps, crossings and, especially, transitions between conductor rail and conventional catenary, since the use of overhead conductor rail is limited to tunnels, as already mentioned. This paper describes simulation techniques developed in order to take into account these critical zones. Furthermore, some specific simulations results are presented that have been used to analyze and optimizes the geometry of this special zones to get a better current collection quality, in a real suburban network. This paper presents the work undertaken by the Railways Technology Research Centre (CITEF), having over 10 years of experience in railways research [1-4].
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In this paper we propose an innovative approach to tackle the problem of traffic sign detection using a computer vision algorithm and taking into account real-time operation constraints, trying to establish intelligent strategies to simplify as much as possible the algorithm complexity and to speed up the process. Firstly, a set of candidates is generated according to a color segmentation stage, followed by a region analysis strategy, where spatial characteristic of previously detected objects are taken into account. Finally, temporal coherence is introduced by means of a tracking scheme, performed using a Kalman filter for each potential candidate. Taking into consideration time constraints, efficiency is achieved two-fold: on the one side, a multi-resolution strategy is adopted for segmentation, where global operation will be applied only to low-resolution images, increasing the resolution to the maximum only when a potential road sign is being tracked. On the other side, we take advantage of the expected spacing between traffic signs. Namely, the tracking of objects of interest allows to generate inhibition areas, which are those ones where no new traffic signs are expected to appear due to the existence of a TS in the neighborhood. The proposed solution has been tested with real sequences in both urban areas and highways, and proved to achieve higher computational efficiency, especially as a result of the multi-resolution approach.
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We have used electron cryo-microscopy and image analysis to examine the native structure of immature, protease-deficient (PR−) and mature, wild-type (WT) Moloney murine leukemia virus (MuLV). Maturational cleavage of the Gag polyprotein by the viral protease is associated with striking morphological changes. The PR− MuLV particles exhibit a rounded central core, which has a characteristic track-like shell on its surface, whereas the WT MuLV cores display a polygonal surface with loss of the track-like feature. The pleomorphic shape and inability to refine unique orientation angles suggest that neither the PR− nor the WT MuLV adheres to strict icosahedral symmetry. Nevertheless, the PR− MuLV particles do exhibit paracrystalline order with a spacing between Gag molecules of ≈45 Å and a length of ≈200 Å. Because of the pleomorphic shape and paracrystalline packing of the Gag–RNA complexes, we raise the possibility that assembly of MuLV is driven by protein–RNA, as well as protein–protein, interactions. The maturation process involves a dramatic reorganization of the packing arrangements within the ribonucleoprotein core with disordering and loosening of the individual protein components.
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The function of repressor activator protein 1 (Rap1p) at glycolytic enzyme gene upstream activating sequence (UAS) elements in Saccharomyces cerevisiae is to facilitate binding of glycolysis regulatory protein 1 (Gcr1p) at adjacent sites. Rap1p has a modular domain structure. In its amino terminus there is an asymmetric DNA-bending domain, which is distinct from its DNA-binding domain, which resides in the middle of the protein. In the carboxyl terminus of Rap1p lie its silencing and putative activation domains. We carried out a molecular dissection of Rap1p to identify domains contributing to its ability to facilitate binding of Gcr1p. We prepared full-length and three truncated versions of Rap1p and tested their ability to facilitate binding of Gcr1p by gel shift assay. The ability to detect ternary complexes containing Rap1p⋅DNA⋅Gcr1p depended on the presence of binding sites for both proteins in the probe DNA. The DNA-binding domain of Rap1p, although competent to bind DNA, was unable to facilitate binding of Gcr1p. Full-length Rap1p and the amino- and carboxyl-truncated versions of Rap1p were each able to facilitate binding of Gcr1p at an appropriately spaced binding site. Under these conditions, Gcr1p displayed an approximately 4-fold greater affinity for Rap1p-bound DNA than for otherwise identical free DNA. When spacing between Rap1p- and Gcr1p-binding sites was altered by insertion of five nucleotides, the ability to form ternary Rap1p⋅DNA⋅Gcr1p complexes was inhibited by all but the DNA-binding domain of Rap1p itself; however, the ability of each individual protein to bind the DNA probe was unaffected.
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The trp RNA-binding attenuation protein of Bacillus subtilis, TRAP, regulates both transcription and translation by binding to specific transcript sequences. The optimal transcript sequences required for TRAP binding were determined by measuring complex formation between purified TRAP protein and synthetic RNAs. RNAs were tested that contained repeats of different trinucleotide sequences, with differing spacing between the repeats. A transcript containing GAG repeats separated by two-nucleotide spacers was bound most tightly. In addition, transmission electron microscopy was used to examine the structure of TRAP and the TRAP-transcript complex. TRAP was observed to be a toroid-shaped oligomer when free or when bound to either a natural or a synthetic RNA.
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A dinâmica ambiental possui capacidade limitada de reciclagem e a crescente utilização resíduos agroindustriais, especialmente na agricultura, pode levar a situações de poluição do solo e demais componentes ambientais. A manutenção da produtividade de ecossistemas agrícolas e naturais depende do processo de transformação da matéria orgânica e, por conseguinte, da biomassa microbiana do solo, e que é responsável pela decomposição e mineralização de resíduos no mesmo. A dinâmica natural dos microrganismos do solo, em constante mudança e adaptação, os torna um indicador sensível às mudanças resultantes de diferentes práticas de manejo agrícola. Sendo assim, conhecer essas alterações e suas interferências é fundamental para identificar estratégias adequadas de manejo, apontando técnicas de utilização adequadas. O objetivo deste trabalho foi avaliar a qualidade de um solo agrícola, cultivado com três variedades de cana-de-açúcar (Saccharum spp.), comparando a utilização de adubação mineral frente à utilização de fertilizante orgânico composto no período final de formação dos perfilhos (120 dias após o plantio). Foi implantado, em condições de campo, o cultivo de cana-de-açúcar (cana planta), utilizando as variedades RB 867515, RB 962869 e RB 855453, onde cada variedade foi cultivada de três formas distintas, sendo elas: plantio controle (CT) sem aplicação de insumos para adubação; plantio orgânico (OG) com aplicação de fertilizante orgânico; e plantio convencional (CV) com aplicação de adubação mineral, seguindo recomendações de adubação após análise química inicial do solo local. Cada parcela possuía 37 m2, com 5 sulcos de 5,0 m de comprimento e espaçamento de 1,5 m entrelinhas, sendo os três sulcos centrais formando a área útil. De acordo com a variedade e o tipo de adubação, foram formados nove tratamentos: T1 86CT, T2 96CT, T3 85CT, T4 6OG, T5 96OG, T6 85OG, T7 86CV, T8 96CV e T9 85CV, com delineamento estatístico de blocos ao acaso e quatro repetições. Os parâmetros químicos do solo analisados foram macronutrientes e micronutrientes; os parâmetros microbiológicos foram carbono da biomassa microbiana (CBM), respiração basal do solo (RBS), quociente metabólico (qCO2), número mais provável de fungos e bactérias do solo (NMP); e, por fim, a produtividade agrícola (t/ha). Os resultados foram submetidos a análise de variância (ANOVA) e à comparação das médias através do teste de Tukey (10%). Também foi realizada a análise de variância dos dados e correlação cofenética de Pearson para formação de dendogramas. Com base no período estudado, considerado como fase crítica da formação do canavial, concluiu-se que os parâmetros químicos que evidenciaram alterações no solo foram pH e os macronutrientes Mg, Al e SB, sendo os tratamentos orgânicos equivalentes e/ou melhores que os tratamentos convencionais. Para os parâmetros microbiológicos, o NMP de fungos apresentou os maiores valores nos tratamentos convencionais e controle. A produtividade agrícola não foi influenciada pelos diferentes tratamentos e insumos utilizados, independente da variedade de cana-de-açúcar utilizada. Por fim, foram observadas correlações positivas entre as variáveis CTC e quociente metabólico (qCO2) apontando potencial melhoria da qualidade do solo, com o emprego de insumos orgânicos
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O presente estudo foi dividido em três capitulos, todos realizados na Estação Experimental de Ciências Florestais de Anhembi/SP, entre os anos de 2014 e 2015. O primeiro estudo intitulado de \"Variação mensal da fitomassa da forragem em função do grau de cobertura do dossel em sistemas silvipastoris\", foi realizado em 3 monoculturas de 13 anos de idade, com área útil de 50 m x 30 m para coleta de pasto as mesmas efectuadas mensalmente. Os resultados apresentaram que não há relação significativa entre a cobertura do dossel e fitomassa da forragem pelo caso de que o sub-bosque estava muito sombreado. Entretanto, houve uma relação indireta entre área basal e fitomassa. Evidenciando-se que o talhão de Eucalipto urograndis apresentou as melhores condições de crescimento e disponibilidade de materia seca mensal para Bachiaria decumbens além de obter a maior porcentagem de folha entre todos os tratamentos. Ao contrario, no talhão de Pinus tecunumanii, foi encontrada a menor disponibilidade de materia seca mensal e por consequência, menor porcentagem de folha. O segundo estudo foi chamado de: \"Disponibilidade de fitomassa de B. decumbens, em um sistema silvipastoril com eucalipto: o papel da radiação\" onde o componente florestal foi o eucalyptus (COP-1377) de 2 anos de idade plantado em uma área útil de 10 ha, dividido em 3 tratamentos (onda longa-OL (39 m), onda curta-OC (21 m), e testemunha-T) e instalado em 4 blocos distintos. Foram realizadas duas coletas dutante o período de verão e de inverno, onde foi possível verificar que o tratamento OL mostrou maior disponibilidade de fitomassa a 65% de irradiância além de obter maior porcentagem da fração folha. Este foi favorecido pelo maior espaçamento entre as aléias. Contudo, houve ataque de cigarinha na pastagem, mantendo a queda da disponibilidade no período de inverno. O terceiro estudo intitulado de: \"Variaçoes arquiteturais de uma monocultura de E. urograndis em função de sua posição espacial\", foi também realizado na monocultura do primeiro estudo, numa área de 7 ha. Para este estudo, realizou-se um inventario florestal, logo após, dividiu-se as árvores por sua classe diamétrica e selecionou-se aleatoriamente 60 árvores para cubagem, e destas, escolheu-se 15 para determinação da fitomassa e respectiva densidade da madeira. Para a obtenção da fitomassa dividiu-se as árvores em três frações de análise: tronco, galhos e folha. Além disso, as 15 árvores foram divididas em: bordadura, intermediária e centro da parcela, de acordo com a sua localização. Verificou-se que a bordadura apresentou os maiores crescimentos em DAP, altura, largura de copa e, que por consequência, obteve maior volume e fitomassa em todas suas frações. Também foi possível observar que tanto a bordadura quanto o centro apresentaram maior densidade básica em função da maior copa e altura das árvores incentivando a geração de mais fitomassa foliar. Finalmente conforme os três estudos realizados neste trabalho de pesquisa, concluiu-se que a radiação solar é fator chave na produtividade da cultura forrageira, demonstrando a necessidade de mais pesquisas sobre os sistemas agroforestais e silvipastoris para o sucesso de futuros emprendimentos.
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Les produits biologiques représentent une avenue thérapeutique très prometteuse pour diverses maladies actuellement sans traitement, dont le cancer. La demande pour ces produits est donc très forte et des bioprocédés industriels efficaces et fiables doivent être mis en place pour y répondre. Le système inductible au cumate (CR5) développé par le groupe de Bernard Massie permet d’exprimer des protéines d’intérêt de façon finement régulable et à haut niveau dans les cellules CHO. Un travail d’optimisation est toutefois nécessaire afin de maximiser l’expression tout en améliorant l’étanchéité du système. Dans cette optique, diverses constructions du promoteur comportant des configurations différentes d’espacement entre ses constituants, des transactivateurs comportant des domaines d’activation différents, et une séquence opératrice synthétique ont été testées pour évaluer leur capacité à améliorer le rendement et l’étanchéité du CR5. Ainsi, un protomoteur comportant trois séquences opératrices avec six paires de bases entre chacune de ces dernières s’est montré plus efficace en termes de rendement et d’étanchéité que la configuration actuelle du CR5. De plus, une nouvelle configuration du CR5 où le transactivateur est régulé par le système inductible à la coumermycine a été étudiée et a montré une régulation très fine. Le travail d’optimisation effectué dans ce projet s’applique seulement dans le but d’optimiser un procédé dans des conditions spécifiques. Son application à d’autres lignées cellulaires et d’autres promoteurs reste à démontrer.
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Analogue model experiments using both brittle and viscous materials were performed to investigate the development and interaction of strike-slip faults in zones of distributed shear deformation. At low strain, bulk dextral shear deformation of an initial rectangular model is dominantly accommodated by left-stepping, en echelon strike-slip faults (Riedel shears, R) that form in response to the regional (bulk) stress field. Push-up zones form in the area of interaction between adjacent left-stepping Riedel shears. In cross sections, faults bounding push-up zones have an arcuate shape or merge at depth. Adjacent left-stepping R shears merge by sideways propagation or link by short synthetic shears that strike subparallel to the bulk shear direction. Coalescence of en echelon R shears results in major, through-going faults zones (master faults). Several parallel master faults develop due to the distributed nature of deformation. Spacing between master faults is related to the thickness of the brittle layers overlying the basal viscous layer. Master faults control to a large extent the subsequent fault pattern. With increasing strain, relatively short antithetic and synthetic faults develop mostly between old, but still active master faults. The orientation and evolution of the new faults indicate local modifications of the stress field. In experiments lacking lateral borders, closely spaced parallel antithetic faults (cross faults) define blocks that undergo clockwise rotation about a vertical axis with continuing deformation. Fault development and fault interaction at different stages of shear strain in our models show similarities with natural examples that have undergone distributed shear.