1000 resultados para DYNAMIC LOCALIZATION


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We present evidence that a bacterial signal transduction cascade that couples morphogenesis with cell cycle progression is regulated by dynamic localization of its components. Previous studies have implicated two histidine kinases, DivJ and PleC, and the response regulator, DivK, in the regulation of morphogenesis in the dimorphic bacterium Caulobacter crescentus. Here, we show that the cytoplasmic response regulator, DivK, exhibits a dynamic, cyclical localization that culminates in asymmetric distribution of DivK within the two cell types that are characteristic of the Caulobacter cell cycle; DivK is dispersed throughout the cytoplasm of the progeny swarmer cell and is localized to the pole of the stalked cell. The membrane-bound DivJ and PleC histidine kinases, which are asymmetrically localized at the opposite poles of the predivisional cell, control the temporal and spatial localization of DivK. DivJ mediates DivK targeting to the poles whereas PleC controls its release from one of the poles at times and places that are consistent with the activities and location of DivJ and PleC in the late predivisional cell. Thus, dynamic changes in subcellular location of multiple components of a signal transduction cascade may constitute a novel mode of prokaryotic regulation to generate and maintain cellular asymmetry.

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In Saccharomyces cerevisiae, entry into mitosis requires activation of the cyclin-dependent kinase Cdc28 in its cyclin B (Clb)-associated form. Clb-bound Cdc28 is susceptible to inhibitory tyrosine phosphorylation by Swe1 protein kinase. Swe1 is itself negatively regulated by Hsl1, a Nim1-related protein kinase, and by Hsl7, a presumptive protein-arginine methyltransferase. In vivo all three proteins localize to the bud neck in a septin-dependent manner, consistent with our previous proposal that formation of Hsl1-Hsl7-Swe1 complexes constitutes a checkpoint that monitors septin assembly. We show here that Hsl7 is phosphorylated by Hsl1 in immune-complex kinase assays and can physically associate in vitro with either Hsl1 or Swe1 in the absence of any other yeast proteins. With the use of both the two-hybrid method and in vitro binding assays, we found that Hsl7 contains distinct binding sites for Hsl1 and Swe1. A differential interaction trap approach was used to isolate four single-site substitution mutations in Hsl7, which cluster within a discrete region of its N-terminal domain, that are specifically defective in binding Hsl1. When expressed in hsl7Δ cells, each of these Hsl7 point mutants is unable to localize at the bud neck and cannot mediate down-regulation of Swe1, but retains other functions of Hsl7, including oligomerization and association with Swe1. GFP-fusions of these Hsl1-binding defective Hsl7 proteins localize as a bright perinuclear dot, but never localize to the bud neck; likewise, in hsl1Δ cells, a GFP-fusion to wild-type Hsl7 or native Hsl7 localizes to this dot. Cell synchronization studies showed that, normally, Hsl7 localizes to the dot, but only in cells in the G1 phase of the cell cycle. Immunofluorescence analysis and immunoelectron microscopy established that the dot corresponds to the outer plaque of the spindle pole body (SPB). These data demonstrate that association between Hsl1 and Hsl7 at the bud neck is required to alleviate Swe1-imposed G2-M delay. Hsl7 localization at the SPB during G1 may play some additional role in fine-tuning the coordination between nuclear and cortical events before mitosis.

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We study the dynamics of a one-dimensional lattice model of hard core bosons which is initially in a superfluid phase with a current being induced by applying a twist at the boundary. Subsequently, the twist is removed, and the system is subjected to periodic delta-function kicks in the staggered on-site potential. We present analytical expressions for the current and work done in the limit of an infinite number of kicks. Using these, we show that the current (work done) exhibits a number of dips (peaks) as a function of the driving frequency and eventually saturates to zero (a finite value) in the limit of large frequency. The vanishing of the current (and the saturation of the work done) can be attributed to a dynamic localization of the hard core bosons occurring as a consequence of the periodic driving. Remarkably, we show that for some specific values of the driving amplitude, the localization occurs for any value of the driving frequency. Moreover, starting from a half-filled lattice of hard core bosons with the particles localized in the central region, we show that the spreading of the particles occurs in a light-cone-like region with a group velocity that vanishes when the system is dynamically localized.

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The dynamic localization of saturated soil is investigated by considering the influence of higher strain gradient. It is shown that the strain gradient has a significant influence on the evolution of shear band in saturated soil and that the width of shear band is proportional to the square root of the strain gradient softening coefficient. The numerical simulation is processed to investigate the influences of shear strain gradient and other factors on the evolution of shear band.

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Within the one-dimensional tight-binding model;rnd chi-3 approximation, we have calculated four-wave-mixing (FWM) signals for a semiconductor superlattice in the presence of both static and high-frequency electric fields. When the exciton effect is negligible, the time-periodic field dynamically delocalizes the otherwise localized Wannier-Stark states, and accordingly quasienergy band structures are formed, and manifest in the FWM spectra as a series of equally separated continua. The width of each continuum is proportional to the joint width of the valence and conduction minibands and is independent of the Wannier-Stark index. The realistic homogeneous broadening blurs the continua into broad peaks, whose line shapes, far from the Lorentzian, vary with the delay time in the FWM spectra. The swinging range of the peaks is just the quasienergy bandwidth. The dynamical delocalization (DDL) also induces significant FWM signals well beyond the excitation energy window. When the Coulomb interaction is taken into account, the unequal spacing between the excitonic Wannier-Stark levels weakens the DDL effect, and the FWM spectrum is transformed into groups of discrete lines. Strikingly, the groups are evenly spaced by the ac field frequency, reflecting the characteristic of the quasienergy states. The homogeneous broadening again smears out the line structures, leading to the excitonic FWM spectra quite similar to those without the exciton effect. However, all these features predicted by the dynamical theory do not appear in a recent experiment [Phys. Rev. Lett. 79, 301 (1997)], in which, by using the static approximation the observed Wannier-Stark ladder with delay-time-dependent spacing in the FWM spectra is attributed to a temporally periodic dipole field, produced by the Bloch oscillation of electrons in real space. The contradiction between the dynamical theory and the experiments is discussed. In addition, our calculation indicates that the dynamical localization coherently enhances the time-integrated FWM signals. The feasibility of using such a technique to study the dynamical localization phenomena is shown. [S0163-1829(99)10607-6].

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Pós-graduação em Matematica Aplicada e Computacional - FCT

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Cardiolipin (CL) plays a key role in dynamic organization of bacterial and mitochondrial membranes. CL forms membrane domains in bacterial cells, and these domains appear to participate in binding and functional regulation of multi-protein complexes involved in diverse cellular functions including cell division, energy metabolism, and membrane transport. Visualization of CL domains in bacterial cells by the fluorescent dye 10-N-nonyl acridine orange is critically reviewed. Possible mechanisms proposed for CL dynamic localization in bacterial cells are discussed. In the mitochondrial membrane CL is involved in organization of multi-subunit oxidative phosphorylation complexes and in their association into higher order supercomplexes. Evidence suggesting a possible role for CL in concert with ATP synthase oligomers in establishing mitochondrial cristae morphology is presented. Hypotheses on CL-dependent dynamic re-organization of the respiratory chain in response to changes in metabolic states and CL dynamic re-localization in mitochondria during the apoptotic response are briefly addressed.

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The cytoskeleton plays an important role in neuronal morphogenesis. We have identified and characterized a novel actin-binding protein, termed Mayven, predominantly expressed in brain. Mayven contains a BTB (broad complex, tramtrack, bric-a-brac)/POZ (poxvirus, zinc finger) domain-like structure in the predicted N terminus and “kelch repeats” in the predicted C-terminal domain. Mayven shares 63% identity (77% similarity) with the Drosophila ring canal (“kelch”) protein. Somatic cell-hybrid analysis indicated that the human Mayven gene is located on chromosome 4q21.2, whereas the murine homolog gene is located on chromosome 8. The BTB/POZ domain of Mayven can self-dimerize in vitro, which might be important for its interaction with other BTB/POZ-containing proteins. Confocal microscopic studies of endogenous Mayven protein revealed a highly dynamic localization pattern of the protein. In U373-MG astrocytoma/glioblastoma cells, Mayven colocalized with actin filaments in stress fibers and in patchy cortical actin-rich regions of the cell margins. In primary rat hippocampal neurons, Mayven is highly expressed in the cell body and in neurite processes. Binding assays and far Western blotting analysis demonstrated association of Mayven with actin. This association is mediated through the “kelch repeats” within the C terminus of Mayven. Depolarization of primary hippocampal neurons with KCl enhanced the association of Mayven with actin. This increased association resulted in dynamic changes in Mayven distribution from uniform to punctate localization along neuronal processes. These results suggest that Mayven functions as an actin-binding protein that may be translocated along axonal processes and might be involved in the dynamic organization of the actin cytoskeleton in brain cells.

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The paper focuses on reliability based design of bridge abutments when subjected to earthquake loading. Planar failure surface has been used in conjunction with pseudo-dynamic approach to compute the seismic active earth pressures on the bridge abutment. The proposed pseudo dynamic method, considers the effects of strain localization in the backfill soil and associated post-peak reduction in the shear resistance from peak to residual values along a previously formed failure plane, phase difference in shear waves and soil amplification along with the horizontal seismic accelerations. Four modes of stability viz. sliding, overturning, eccentricity and bearing capacity of the foundation soil are considered for the reliability analysis. The influence of various design parameters on the seismic reliability indices against four modes of failure is presented, following the suggestions of Japan Road Association, Caltrans Bridge Design Specifications and U.S Department of the Army.

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The microstructural evolution in localized shear deformation was investigated in an 8090 Al-Li alloy by split Hopkinson pressure bar (strain rate of approximately 10(3) s(-1)) at ambient temperature and 77 K. The alloy was tested in the peak-, over-, under-, and natural-aged conditions, that provide a wide range of microstructural parameters and mechanical properties. Two types of localized shear bands were distinguished by optical microscopy: the deformed shear band and the white-etching shear band. They form at different stages of deformation during localization. There are critical strains for the occurrence of deformed and white-etching localized shear deformation, at the imposed strain rate. Observations by transmission electron microscopy reveal that the white-etching bands contain fine equiaxed grains; it is proposed that they are the result of recrystallization occurring during localization. The deformed-type bands are observed after testing at 77 K in all heat treatment conditions, but they are not as well defined as those developed at ambient temperature. Cracking often occurs along the localized shear at ambient temperature. The decrement in temperature is favorable for the nucleation, growth and coalescence of the microcracks along the shear bands, inducing fracture.

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Investigations made by the authors and collaborators into the microstructural aspects of adiabatic shear localization are critically reviewed. The materials analyzed are low-carbon steels, 304 stainless steel, monocrystalline Fe-Ni-Cr, Ti and its alloys, Al-Li alloys, Zircaloy, copper, and Al/SiCp composites. The principal findings are the following: (a) there is a strain-rate-dependent critical strain for the development of shear bands; (b) deformed bands and white-etching bands correspond to different stages of deformation; (c) different slip activities occur in different stages of band development; (d) grain refinement and amorphization occur in shear bands; (e) loss of stress-carrying capability is more closely associated with microdefects rather than with localization of strain; (f) both crystalline rotation and slip play important roles; and (g) band development and band structures are material dependent. Additionally, avenues for new research directions are suggested.

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The localized shear deformation in the 2024 and 2124 Al matrix composites reinforced with SiC particles was investigated with a split Hopkinson pressure bar (SHPB) at a strain rate of about 2.0x10(3) s(-1). The results showed that the occurrence of localized shear deformation is sensitive to the size of SiC particles. It was found that the critical strain, at which the shear localization occurs, strongly depends on the size and volume fraction of SiC particles. The smaller the particle size, the lower the critical strain required for the shear localization. TEM examinations revealed that Al/SiCp interfaces are the main sources of dislocations. The dislocation density near the interface was found to be high and it decreases with the distance from the particles. The Al matrix in shear bands was highly deformed and severely elongated at low angle boundaries. The Al/SiCp interfaces, particularly the sharp corners of SiC particles, provide the sites for microcrack initiation. Eventual fracture is caused by the growth and coalescence of microcracks along the shear bands. It is proposed that the distortion free equiaxed grains with low dislocation density observed in the center of shear band result from recrystallization during dynamic deformation.

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A framework for the simultaneous localization and recognition of dynamic hand gestures is proposed. At the core of this framework is a dynamic space-time warping (DSTW) algorithm, that aligns a pair of query and model gestures in both space and time. For every frame of the query sequence, feature detectors generate multiple hand region candidates. Dynamic programming is then used to compute both a global matching cost, which is used to recognize the query gesture, and a warping path, which aligns the query and model sequences in time, and also finds the best hand candidate region in every query frame. The proposed framework includes translation invariant recognition of gestures, a desirable property for many HCI systems. The performance of the approach is evaluated on a dataset of hand signed digits gestured by people wearing short sleeve shirts, in front of a background containing other non-hand skin-colored objects. The algorithm simultaneously localizes the gesturing hand and recognizes the hand-signed digit. Although DSTW is illustrated in a gesture recognition setting, the proposed algorithm is a general method for matching time series, that allows for multiple candidate feature vectors to be extracted at each time step.

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Received signal strength-based localization systems usually rely on a calibration process that aims at characterizing the propagation channel. However, due to the changing environmental dynamics, the behavior of the channel may change after some time, thus, recalibration processes are necessary to maintain the positioning accuracy. This paper proposes a dynamic calibration method to initially calibrate and subsequently update the parameters of the propagation channel model using a Least Mean Squares approach. The method assumes that each anchor node in the localization infrastructure is characterized by its own propagation channel model. In practice, a set of sniffers is used to collect RSS samples, which will be used to automatically calibrate each channel model by iteratively minimizing the positioning error. The proposed method is validated through numerical simulation, showing that the positioning error of the mobile nodes is effectively reduced. Furthermore, the method has a very low computational cost; therefore it can be used in real-time operation for wireless resource-constrained nodes.