486 research outputs found

    On a (β-) roll

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    Non-Linear Optimization Applied to Angle-of-Arrival Satellite-Based Geolocation with Correlated Measurements

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    A common remote sensing application is producing geolocation estimates for an object of interest from multiple sensor platforms. Geolocation estimates are desired to help improve situational awareness when dealing with space objects that do not actively broadcast their location. A depiction of the error parameters are calculated in conjunction with the positional estimates. Problems occur when multiple measurements from a single sensor are used to estimate a location due to correlations in sensor error. A non-linear optimization approach is presented for determining geolocation estimates and their associated error parameters. The error parameters directly reflect the error present on the individual measurements used to produce the position estimates. Correlations in errors are dealt with by augmenting the non-linear optimization with a covariance intersection algorithm. Finally, the ability to account for correlated errors within the optimization algorithm is analyzed using Monte-Carlo simulations. The ability to describe an objects location with a given confidence helps aid in the analysis of the system at large

    Heat shock protein expression analysis in canine osteosarcoma reveals HSP60 as a potentially relevant therapeutic target

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    Heat shock proteins (HSP) are highly conserved across eukaryotic and prokaryotic species. These proteins play a role in response to cellular stressors, protecting cells from damage and facilitating recovery. In tumor cells, HSPs can have cytoprotective effects and interfere with apoptotic cascades. This study was performed to assess the prognostic and predictive values of the gene expression of HSP family members in canine osteosarcoma (OS) and their potential for targeted therapy. Gene expressions for HSP were assessed using quantitative PCR (qPCR) on 58 snap-frozen primary canine OS tumors and related to clinic-pathological parameters. A significant increased expression of HSP60 was found in relation to shorter overall survival and an osteoblastic phenotype. Therefore, the function of HSP60 was investigated in more detail. Immunohistochemical analysis revealed heterogeneous staining for HSP60 in tumors. The highest immunoreactivity was found in tumors of short surviving dogs. Next HSP expression was shown in a variety of canine and human OS cell lines by qPCR and Western blot. In two highly metastatic cell lines HSP60 expression was silenced using siRNA resulting in decreased cell proliferation and induction of apoptosis in both cell lines. It is concluded that overexpression of HSP60 is associated with a poor prognosis of OS and should be evaluated as a new target for therapy

    Repositioning the Catalytic Triad Aspartic Acid of Haloalkane Dehalogenase: Effects on Stability, Kinetics, and Structure

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    Haloalkane dehalogenase (DhlA) catalyzes the hydrolysis of haloalkanes via an alkyl-enzyme intermediate. The covalent intermediate, which is formed by nucleophilic substitution with Asp124, is hydrolyzed by a water molecule that is activated by His289. The role of Asp260, which is the third member of the catalytic triad, was studied by site-directed mutagenesis. Mutation of Asp260 to asparagine resulted in a catalytically inactive D260N mutant, which demonstrates that the triad acid Asp260 is essential for dehalogenase activity. Furthermore, Asp260 has an important structural role, since the D260N enzyme accumulated mainly in inclusion bodies during expression, and neither substrate nor product could bind in the active-site cavity. Activity for brominated substrates was restored to D260N by replacing Asn148 with an aspartic or glutamic acid. Both double mutants D260N+N148D and D260N+N148E had a 10-fold reduced kcat and 40-fold higher Km values for 1,2-dibromoethane compared to the wild-type enzyme. Pre-steady-state kinetic analysis of the D260N+N148E double mutant showed that the decrease in kcat was mainly caused by a 220-fold reduction of the rate of carbon-bromine bond cleavage and a 10-fold decrease in the rate of hydrolysis of the alkyl-enzyme intermediate. On the other hand, bromide was released 12-fold faster and via a different pathway than in the wild-type enzyme. Molecular modeling of the mutant showed that Glu148 indeed could take over the interaction with His289 and that there was a change in charge distribution in the tunnel region that connects the active site with the solvent. On the basis of primary structure similarity between DhlA and other α/β-hydrolase fold dehalogenases, we propose that a conserved acidic residue at the equivalent position of Asn148 in DhlA is the third catalytic triad residue in the latter enzymes.

    Діагностика банкрутства в системі антикризового управління

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    Розглянуто питання відповідності і значимості діагностики фінансового стану кожної стадії антикризового управління. Діагностику банкрутства визначено як ретроспективне, оперативне і перспективне дослідження господарської діяльності та процесу управління підприємством, направлене на виявлення параметрів кризового розвитку за допомогою реалізації комплексу дослідницьких процедур, виявлення слабких ланок і "вузьких місць".Рассмотрены вопросы соответствия и значимости диагностики каждой стадии антикризисного управления. Диагностика банкротства определена как ретроспективное, оперативное и перспективное исследование хозяйственной деятельности и процесса управления предприятием, направленное на выявление параметров кризисного развития с помощью реализации комплекса исследовательских процедур, выявления слабых зон и «узких мест».The article analyzes issues of adequacy and role of diagnostics in all stage of anti-crisis management. The diagnostics of bankruptcy is defined as a retrospective, operative and prospective study of enterprise’s economic activity and management process that is directed to identification of parameters of crisis development by using comprehensive researches and to finding weak spots and "bottlenecks"

    Structure of maltoheptaose by difference Fourier methods and a model for glycogen

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    The structure of the glucose heptamer, maltoheptaose, has been determined by difference Fourier analysis at 0.25 nm resolution through its binding to phosphorylase a. It is a left-handed helical structure, with 6.5 glucose residues per turn and a rise per residue of 2.4 Å. The molecule shows short-range order when no protein is present to stabilize its conformation in solution. With one exception, the individual torsion angles between sugar residues vary over a narrow range and preserve a good O(2)O(3′) hydrogen bond. The length of an individual chain for glycogen can be extrapolated from the maltoheptaose data and agrees well with the size for glycogen predicted by the Whelan model

    Scene Decomposed Blind Deconvolution and Neural Network based Multi-Frame Image Restoration Techniques for Astronomical Imagery

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    Ground based astronomical imaging is an important method in gaining situational awareness of orbiting and far off objects in space. This method of imaging is accessible to everyone that can look up into the sky, but the accessibility to digital telescope systems allows for more exciting methods of extracting information. A use case for these telescopes is finding nearby objects to larger brighter known objects. The number satellites in low-earth orbit and geosynchronous earth orbit is becoming more congested as these orbits increase in population. Tens of thousands of satellites and debris now exist in this orbit, with the number expected to grow. This drives a need to know the proximity of potential unknown objects to known assets. Using larger telescopes increases the number of photons a camera can collect, giving us the ability to peer further into space or conversely find fainter objects nearby. This paper focuses on the algorithmic methods of image restoration and detection of objects within ground based imagery, seeking to quantify statistical and neural network methods that perform these tasks

    The coordination of cell growth during fission yeast mating requires Ras1-GTP hydrolysis

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    The spatial and temporal control of polarity is fundamental to the survival of all organisms. Cells define their polarity using highly conserved mechanisms that frequently rely upon the action of small GTPases, such as Ras and Cdc42. Schizosaccharomyces pombe is an ideal system with which to study the control of cell polarity since it grows from defined tips using Cdc42-mediated actin remodeling. Here we have investigated the importance of Ras1-GTPase activity for the coordination of polarized cell growth during fission yeast mating. Following pheromone stimulation, Ras1 regulates both a MAPK cascade and the activity of Cdc42 to enable uni-directional cell growth towards a potential mating partner. Like all GTPases, when bound to GTP, Ras1 adopts an active conformation returning to an inactive state upon GTP-hydrolysis, a process accelerated through interaction with negative regulators such as GAPs. Here we show that, at low levels of pheromone stimulation, loss of negative regulation of Ras1 increases signal transduction via the MAPK cascade. However, at the higher concentrations observed during mating, hyperactive Ras1 mutations promote cell death. We demonstrate that these cells die due to their failure to coordinate active Cdc42 into a single growth zone resulting in disorganized actin deposition and unsustainable elongation from multiple tips. These results provide a striking demonstration that the deactivation stage of Ras signaling is fundamentally important in modulating cell polarity

    Crystallization of the gene 45 protein from the DNA replication fork of bacteriophage T4

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    The gene 45 protein from bacteriophage T4 has been purified and is crystallized. This protein is part of the T4 DNA replication complex. The crystallized protein is active in complementation assays. X-ray diffraction analysis is in progress; data are measured for the native and several heavy atom derivatives. The crystals diffract to about 3.5-Å resolution
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