927 resultados para Yeast Two Hybrid
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An earlier discussion of sp hybrid orbitals is extended to include orbitals made up to an s orbital and two p orbitals.
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In the biomedical studies, the general data structures have been the matched (paired) and unmatched designs. Recently, many researchers are interested in Meta-Analysis to obtain a better understanding from several clinical data of a medical treatment. The hybrid design, which is combined two data structures, may create the fundamental question for statistical methods and the challenges for statistical inferences. The applied methods are depending on the underlying distribution. If the outcomes are normally distributed, we would use the classic paired and two independent sample T-tests on the matched and unmatched cases. If not, we can apply Wilcoxon signed rank and rank sum test on each case. ^ To assess an overall treatment effect on a hybrid design, we can apply the inverse variance weight method used in Meta-Analysis. On the nonparametric case, we can use a test statistic which is combined on two Wilcoxon test statistics. However, these two test statistics are not in same scale. We propose the Hybrid Test Statistic based on the Hodges-Lehmann estimates of the treatment effects, which are medians in the same scale.^ To compare the proposed method, we use the classic meta-analysis T-test statistic on the combined the estimates of the treatment effects from two T-test statistics. Theoretically, the efficiency of two unbiased estimators of a parameter is the ratio of their variances. With the concept of Asymptotic Relative Efficiency (ARE) developed by Pitman, we show ARE of the hybrid test statistic relative to classic meta-analysis T-test statistic using the Hodges-Lemann estimators associated with two test statistics.^ From several simulation studies, we calculate the empirical type I error rate and power of the test statistics. The proposed statistic would provide effective tool to evaluate and understand the treatment effect in various public health studies as well as clinical trials.^
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Cells are exposed to a variety of environmental and physiological changes including temperature, pH and nutrient availability. These changes cause stress to cells, which results in protein misfolding and altered cellular protein homeostasis. How proteins fold into their three-dimensional functional structure is a fundamental biological process with important relevance to human health. Misfolded and aggregated proteins are linked to multiple neurodegenerative diseases, cardiovascular disease and cystic fibrosis. To combat proteotoxic stress, cells deploy an array of molecular chaperones that assist in the repair or removal of misfolded proteins. Hsp70, an evolutionarily conserved molecular chaperone, promotes protein folding and helps maintain them in a functional state. Requisite co-chaperones, including nucleotide exchange factors (NEFs) strictly regulate and serve to recruit Hsp70 to distinct cellular processes or locations. In yeast and human cells, three structurally non-related cytosolic NEFs are present: Sse1 (Hsp110), Fes1 (HspBP1) and Snl1 (Bag-1). Snl1 is unique among the cytosolic NEFs as it is localized at the ER membrane with its Hsp70 binding (BAG) domain exposed to the cytosol. I discovered that Snl1 distinctly interacts with assembled ribosomes and several lines of evidence indicate that this interaction is both independent of and concurrent with binding to Hsp70 and is not dependent on membrane localization. The ribosome-binding site is identified as a short lysine-rich motif within the amino terminus of the Snl1 BAG domain distinct from the Hsp70 interaction region. In addition, I demonstrate ribosome association with the Snl1 homolog in the pathogenic fungus, Candida albicans and localize this putative NEF to a perinuclear/ER membrane, suggesting functional conservation in fungal BAG domain-containing proteins. As a first step in determining specific domain architecture in fungal BAG proteins, I present the preliminary steps of protein purification and analysis of the minimal Hsp70 binding region in in both S.cerevisiae and C. albicans Snl1. Contrary to previous in vitro evidence which showed the Fes1 NEF to interact with both cytosolic Hsp70s, Ssa and Ssb, Fes1 is shown to interact specifically with Ssa when expressed under normal cellular conditions in S. cerevisiae. This is the first reported evidence of Hsp70 binding selectivity for a cytosolic NEF, and suggests a possible mechanism to achieve specificity in Hsp70-dependent functions. Taken together, the work presented in this dissertation highlights the striking divergence among Hsp70 co-chaperones in selecting binding partners, which may correlate with their specific roles in the cell.
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Cells govern their activities and modulate their interactions with the environment to achieve homeostasis. The heat shock response (HSR) is one of the most well studied fundamental cellular responses to environmental and physiological challenges, resulting in rapid synthesis of heat shock proteins (HSPs), which serve to protect cellular constituents from the deleterious effects of stress. In addition to its role in cytoprotection, the HSR also influences lifespan and is associated with a variety of human diseases including cancer, aging and neurodegenerative disorders. In most eukaryotes, the HSR is primarily mediated by the highly conserved transcription factor HSF1, which recognizes target hsp genes by binding to heat shock elements (HSEs) in their promoters. In recent years, significant efforts have been made to identify small molecules as potential pharmacological activators of HSF1 that could be used for therapeutic benefit in the treatment of human diseases relevant to protein conformation. However, the detailed mechanisms through which these molecules drive HSR activation remain unclear. In this work, I utilized the baker's yeast Saccharomyces cerevisiae as a model system to identify a group of thiol-reactive molecules including oxidants, transition metals and metalloids, and electrophiles, as potent activators of yeast Hsf1. Using an artificial HSE-lacZ reporter and the glucocorticoid receptor system (GR), these diverse thiol-reactive compounds are shown to activate Hsf1 and inhibit Hsp90 chaperone complex activity in a reciprocal, dose-dependent manner. To further understand whether cells sense these reactive compounds through accumulation of unfolded proteins, the proline analog azetidine-2-carboxylic acid (AZC) and protein cross-linker dithiobis(succinimidyl propionate) (DSP) were used to force misfolding of nascent polypeptides and existing cytosolic proteins, respectively. Both unfolding reagents display kinetic HSP induction profiles dissimilar to those generated by thiol-reactive compounds. Moreover, AZC treatment leads to significant cytotoxicity, which is not observed in the presence of the thiol-reactive compounds at the concentrations sufficient to induce Hsf1. Additionally, DSP treatment has little to no effect on Hsp90 functions. Together with the ultracentrifugation analysis of cell lysates that detected no insoluble protein aggregates, my data suggest that at concentrations sufficient to induce Hsf1, thiol-reactive compounds do not induce the HSR via a mechanism based on accumulation of unfolded cytosolic proteins. Another possibility is that thiol-reactive compounds may influence aspects of the protein quality control system such as the ubiquitin-proteasome system (UPS). To address this hypothesis, β-galactosidase reporter fusions were used as model substrates to demonstrate that thiol-reactive compounds do not inhibit ubiquitin activating enzymes (E1) or proteasome activity. Therefore, thiol-reactive compounds do not activate the HSR by inhibiting UPS-dependent protein degradation. I therefore hypothesized that these molecules may directly inactivate protein chaperones, known as repressors of Hsf1. To address this possibility, a thiol-reactive biotin probe was used to demonstrate in vitro that the yeast cytosolic Hsp70 Ssa1, which partners with Hsp90 to repress Hsf1, is specifically modified. Strikingly, mutation of conserved cysteine residues in Ssa1 renders cells insensitive to Hsf1 activation by cadmium and celastrol but not by heat shock. Conversely, substitution with the sulfinic acid and steric bulk mimic aspartic acid led to constitutive activation of Hsf1. Cysteine 303, located in the nucleotide-binding/ATPase domain of Ssa1, was shown to be modified in vivo by a model organic electrophile using Click chemistry technology, verifying that Ssa1 is a direct target for thiol-reactive compounds through adduct formation. Consistently, cadmium pretreatment promoted cells thermotolerance, which is abolished in cells carrying SSA1 cysteine mutant alleles. Taken together, these findings demonstrate that Hsp70 acts as a sensor to induce the cytoprotective heat shock response in response to environmental or endogenously produced thiol-reactive molecules and can discriminate between two distinct environmental stressors.
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A method to analyze parabolic reflectors with arbitrary piecewise rim is presented in this communication. This kind of reflectors, when operating as collimators in compact range facilities, needs to be large in terms of wavelength. Their analysis is very inefficient, when it is carried out with fullwave/MoM techniques, and it is not very appropriate for designing with PO techniques. Also, fast GO formulations do not offer enough accuracy to reach performance results. The proposed algorithm is based on a GO-PWS hybrid scheme, using analytical as well as non-analytical formulations. On one side, an analytical treatment of the polygonal rim reflectors is carried out. On the other side, non-analytical calculi are based on efficient operations, such as M2 order 2-dimensional FFT. A combination of these two techniques in the algorithm ensures real ad-hoc design capabilities, reached through analysis speedup. The purpose of the algorithm is to obtain an optimal conformal serrated-edge reflector design through the analysis of the field quality within the quiet zone that it is able to generate in its forward half space.
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In October 2002, under the auspices of Spanish Cooperation, a pilot electrification project put into operation two centralised PV-diesel hybrid systems in two different Moroccan villages. These systems currently provide a full-time energy service and supply electricity to more than a hundred of families, six community buildings, street lighting and one running water system. The appearance of the electricity service is very similar to an urban one: one phase AC supply (230V/50Hz) distributed up to each dwelling using a low-voltage mini-grid, which has been designed to be fully compatible with a future arrival of the utility grid. The management of this electricity service is based on a “fee-for-service” scheme agreed between a local NGO, partner of the project, and electricity associations created in each village, which are in charge of, among other tasks, recording the daily energy production of systems and the monthly energy consumption of each house. This register of data allows a systematic evaluation of both the system performance and the energy consumption of users. Now, after four years of operation, this paper presents the experience of this pilot electrification project and draws lessons that can be useful for designing, managing and sizing this type of small village PV-hybrid system
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Cloud computing is one the most relevant computing paradigms available nowadays. Its adoption has increased during last years due to the large investment and research from business enterprises and academia institutions. Among all the services cloud providers usually offer, Infrastructure as a Service has reached its momentum for solving HPC problems in a more dynamic way without the need of expensive investments. The integration of a large number of providers is a major goal as it enables the improvement of the quality of the selected resources in terms of pricing, speed, redundancy, etc. In this paper, we propose a system architecture, based on semantic solutions, to build an interoperable scheduler for federated clouds that works with several IaaS (Infrastructure as a Service) providers in a uniform way. Based on this architecture we implement a proof-of-concept prototype and test it with two different cloud solutions to provide some experimental results about the viability of our approach.
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The adhesives used for applications in marine environments are subject to particular chemical conditions, which are mainly characterised by an elevated chlorine ion content and intermittent wetting/drying cycles, among others.These conditions can limit the use of adhesives due to the degradation processes that they experience. In this work, the chemical degradation of two different polymers, polyurethane and vinylester, was studied in natural seawater under immersion for different periods of time.The diffusion coefficients and concentration profiles of water throughout the thickness of the adhesiveswere obtained.Microstructural changes in the polymer due to the action of water were observed by SEM, and the chemical degradation of the polymer was monitored with the Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The degradation of the mechanical properties of the adhesive was determined by creep tests withMixed Cantilever Beam (MCB) specimens at different temperatures. After 180 days of immersion of the specimens, it was concluded that the J-integral value (depending on the strain) implies a loss of stiffness of 51% and a decrease in the failure load of 59% for the adhesive tested.
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ATM, SDH or satellite have been used in the last century as the contribution network of Broadcasters. However the attractive price of IP networks is changing the infrastructure of these networks in the last decade. Nowadays, IP networks are widely used, but their characteristics do not offer the level of performance required to carry high quality video under certain circumstances. Data transmission is always subject to errors on line. In the case of streaming, correction is attempted at destination, while on transfer of files, retransmissions of information are conducted and a reliable copy of the file is obtained. In the latter case, reception time is penalized because of the low priority this type of traffic on the networks usually has. While in streaming, image quality is adapted to line speed, and line errors result in a decrease of quality at destination, in the file copy the difference between coding speed vs line speed and errors in transmission are reflected in an increase of transmission time. The way news or audiovisual programs are transferred from a remote office to the production centre depends on the time window and the type of line available; in many cases, it must be done in real time (streaming), with the resulting image degradation. The main purpose of this work is the workflow optimization and the image quality maximization, for that reason a transmission model for multimedia files adapted to JPEG2000, is described based on the combination of advantages of file transmission and those of streaming transmission, putting aside the disadvantages that these models have. The method is based on two patents and consists of the safe transfer of the headers and data considered to be vital for reproduction. Aside, the rest of the data is sent by streaming, being able to carry out recuperation operations and error concealment. Using this model, image quality is maximized according to the time window. In this paper, we will first give a briefest overview of the broadcasters requirements and the solutions with IP networks. We will then focus on a different solution for video file transfer. We will take the example of a broadcast center with mobile units (unidirectional video link) and regional headends (bidirectional link), and we will also present a video file transfer file method that satisfies the broadcaster requirements.
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There is now an emerging need for an efficient modeling strategy to develop a new generation of monitoring systems. One method of approaching the modeling of complex processes is to obtain a global model. It should be able to capture the basic or general behavior of the system, by means of a linear or quadratic regression, and then superimpose a local model on it that can capture the localized nonlinearities of the system. In this paper, a novel method based on a hybrid incremental modeling approach is designed and applied for tool wear detection in turning processes. It involves a two-step iterative process that combines a global model with a local model to take advantage of their underlying, complementary capacities. Thus, the first step constructs a global model using a least squares regression. A local model using the fuzzy k-nearest-neighbors smoothing algorithm is obtained in the second step. A comparative study then demonstrates that the hybrid incremental model provides better error-based performance indices for detecting tool wear than a transductive neurofuzzy model and an inductive neurofuzzy model.
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High power density is strongly preferable for the on-board battery charger of Plug-in Hybrid Electric Vehicle (PHEV). Wide band gap devices, such as Gallium Nitride HEMTs are being explored to push to higher switching frequency and reduce passive component size. In this case, the bulk DC link capacitor of AC-DC Power Factor Correction (PFC) stage, which is usually necessary to store ripple power of two times the line frequency in a DC current charging system, becomes a major barrier on power density. If low frequency ripple is allowed in the battery, the DC link capacitance can be significantly reduced. This paper focuses on the operation of a battery charging system, which is comprised of one Full Bridge (FB) AC-DC stage and one Dual Active Bridge (DAB) DC-DC stage, with charging current containing low frequency ripple at two times line frequency, designated as sinusoidal charging. DAB operation under sinusoidal charging is investigated. Two types of control schemes are proposed and implemented in an experimental prototype. It is proved that closed loop current control is the better. Full system test including both FB AC-DC stage and DAB DC-DC stage verified the concept of sinusoidal charging, which may lead to potentially very high power density battery charger for PHEV.
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Data from an attitudinal survey and stated preference ranking experiment conducted in two urban European interchanges (i.e. City-HUBs) in Madrid (Spain) and Thessaloniki (Greece) show that the importance that City-HUBs users attach to the intermodal infrastructure varies strongly as a function of their perceptions of time spent in the interchange (i.e.intermodal transfer and waiting time). A principal components analysis allocates respondents (i.e. city-HUB users) to two classes with substantially different perceptions of time saving when they make a transfer and of time using during their waiting time.
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In order to achieve to minimize car-based trips, transport planners have been particularly interested in understanding the factors that explain modal choices. In the transport modelling literature there has been an increasing awareness that socioeconomic attributes and quantitative variables are not sufficient to characterize travelers and forecast their travel behavior. Recent studies have also recognized that users? social interactions and land use patterns influence travel behavior, especially when changes to transport systems are introduced, but links between international and Spanish perspectives are rarely deal. In this paper, factorial and path analyses through a Multiple-Indicator Multiple-Cause (MIMIC) model are used to understand and describe the relationship between the different psychological and environmental constructs with social influence and socioeconomic variables. The MIMIC model generates Latent Variables (LVs) to be incorporated sequentially into Discrete Choice Models (DCM) where the levels of service and cost attributes of travel modes are also included directly to measure the effect of the transport policies that have been introduced in Madrid during the last three years in the context of the economic crisis. The data used for this paper are collected from a two panel smartphone-based survey (n=255 and 190 respondents, respectively) of Madrid.
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The deployment of the Ambient Intelligence (AmI) paradigm requires designing and integrating user-centered smart environments to assist people in their daily life activities. This research paper details an integration and validation of multiple heterogeneous sensors with hybrid reasoners that support decision making in order to monitor personal and environmental data at a smart home in a private way. The results innovate on knowledge-based platforms, distributed sensors, connected objects, accessibility and authentication methods to promote independent living for elderly people. TALISMAN+, the AmI framework deployed, integrates four subsystems in the smart home: (i) a mobile biomedical telemonitoring platform to provide elderly patients with continuous disease management; (ii) an integration middleware that allows context capture from heterogeneous sensors to program environment¿s reaction; (iii) a vision system for intelligent monitoring of daily activities in the home; and (iv) an ontologies-based integrated reasoning platform to trigger local actions and manage private information in the smart home. The framework was integrated in two real running environments, the UPM Accessible Digital Home and MetalTIC house, and successfully validated by five experts in home care, elderly people and personal autonomy.
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Nowadays increasing fuel prices and upcoming pollutant emission regulations are becoming a growing concern for the shipping industry worldwide. While fuel prices will keep rising in future years, the new International Convention for the Prevention of Pollution from Ships (MARPOL) and Sulphur Emissions Control Areas (SECA) regulations will forbid ships to use heavy fuel oils at certain situations. To fulfil with these regulations, the next step in the marine shipping business will comprise the use of cleaner fuels on board as well as developing new propulsion concept. In this work a new conceptual marine propulsion system is developed, based on the integration of diesel generators with fuel cells in a 2850 metric tonne of deadweight platform supply vessel. The efficiency of the two 250 kW methanol-fed Solid Oxide Fuel Cell (SOFC) system installed on board combined with the hydro dynamically optimized design of the hull of the ship will allow the ship to successfully operate at certain modes of operation while notably reduce the pollutant emissions to the atmosphere. Besides the cogeneration heat obtained from the fuel cell system will be used to answer different heating needs on board the vessel