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991.
Keetch CA Hernánndez H Sterling A Baumert M Allen MH Robinson CV 《Analytical chemistry》2003,75(18):4937-4941
Nanoflow electrospray mass spectrometry has been applied previously to investigate noncovalent protein-protein and protein-ligand interactions. Here we evaluate a commercial microchip device for this application. We show that the microchip can be used to obtain mass spectra of the noncovalent tetramer transthyretin. The device showed a 10-fold increase in signal stability compared with a nanoflow capillary and a high level of nozzle-to-nozzle reproducibility. Binding of the natural ligand thyroxine was clearly observed, and a range of small molecules proposed as inhibitors of transthyretin amyloidosis were shown to be effective in stabilizing the tetramer. We propose that measuring the ability of small molecules to stabilize protein complexes using this automated microchip technology will enable high-throughput screening of multi-protein complexes by mass spectrometry. 相似文献
992.
Spectroscopic features of dual fluorescence/luminescence resonance energy-transfer molecular beacons 总被引:6,自引:0,他引:6
Molecular beacons have the potential to become a powerful tool in gene detection and quantification in living cells. Here we report a novel dual molecular beacons approach to reduce false-positive signals in detecting target nucleic acids in homogeneous assays. A pair of molecular beacons, each containing a fluorescence quencher and a reporter fluorophore, one with a donor and a second with an acceptor fluorophore, hybridize to adjacent regions on the same target resulting in fluorescence resonance energy transfer (FRET). The detection of a FRET signal leads to a substantially increased signal-to-background ratio compared with that seen in single molecular beacon assays and enables discrimination between fluorescence due to specific probe/target hybridization and a variety of possible false-positive events. Further, when a lanthanide chelate is used as a donor in a dual-probe assay, extremely high signal-to-background ratios can be achieved owing to the long lifetime and sharp emission peaks of the donor and the time-gated detection of acceptor fluorescence emission. These new approaches allow for the ultrasensitive detection of target molecules in a way that could be readily applied to real-time imaging of gene expression in living cells. 相似文献
993.
Wang JS Taylor M Flivik G Lidgren L 《Journal of materials science. Materials in medicine》2003,14(1):55-61
Debonding of the stem–cement interface has been implicated in the initiation of failure of cemented femoral stems. The objective of this work was to examine some of the parameters which influence the interface static shear strength, including surface finish, cement type, pre-treatments and porosity. Surface finish was found to have the greatest effect on the interface strength. Increasing the surface roughness by a factor of 100 increases the interface shear strength by a factor of 20. However, increasing the surface roughness above a certain value was found to have no additional affect. This was due to failure in the cement itself rather than at the cement–stem interface. There were significant differences between some of the different cement types regarding the interface strength. Pre-heating the stem produced a six fold reduction in cement porosity at the stem–cement interface, however, resulting in only a minor influence on the static interface strength. Generally, no significant correlation was found between the cement porosity and the static interfacial shear strength. 相似文献
994.
995.
Staphylococcal enterotoxin A (SEA) cross-links two class II major histocompatibility complex (MHC) molecules and forms a multimeric assembly with T-cell receptors (TcRs). The X-ray crystal structure of SEA has been solved, yet details describing molecular recognition and association remain unclear. We present a structural model for the interactions of SEA with cell-surface proteins. Molecular docking calculations predicting SEA association with the class II MHC molecule HLA-DR1 were performed by using a rigid-body docking method. Docked orientations were evaluated by a Poisson-Boltzmann model for the electrostatic free energy of binding and the hydrophobic effect calculated from molecular surface areas. We found that the best-scoring SEA conformers for the DR1alpha interface display a binding mode similar to that determined crystallographically for staphylococcal enterotoxin B bound to HLA-DR1. For the zinc-binding site of SEA, docking DR1beta yielded several orientations exhibiting tetrahedral-like coordination geometries. Combining the two interfaces, tetramers were modeled by docking an alphabeta TcR with trimolecular complexes DR1beta-SEA-DR1alpha and SEA-betaDR1alpha-SEA. Our results indicate that the complex DR1beta-SEA-DR1alpha provides a more favorable assembly for the engagement of TcRs, forming SEA molecular contacts that are in accord with reported mutagenesis studies. In contrast, the cooperative association of two SEA molecules on a single DR1 molecule sterically inhibits interactions with TcRs. We suggest that signal transduction stimulated by SEA through large-scale assembly is limited to four or five TcR-(DR1beta-SEA-DR1alpha) tetramers and requires the dimerization of class II MHC molecules, while TcR dimerization is unlikely. 相似文献
996.
Open-hole honeycomb sandwich panels with woven graphite/epoxy facesheets and Nomex cores were tested uniaxially in compression to characterize their damage tolerance. A plain weave T-300 graphite fiber fabric was used for the facesheets in two stacking sequences: [45/02] and [03]. Observations of macroscopic sub-critical damage behavior were different in the two material systems. Linear damage zones (LDZ), consisting of fiber micro-buckles and extensive delamination, were typically observed in the [03] material. The [45/02] material exhibited a delamination/bulge zone (DBZ), which consisted of an out-of-plane curved deformation of the outer 45° ply accompanied by a delamination from the interior 0° plies. Modeling of these apparently distinct failure modes, and comparison to experimental data, revealed that the only mode representative of damage tolerant behavior is linear damage zone formation and propagation for both material systems, and that the delamination/bulge behavior is a secondary phenomenon. 相似文献
997.
We advance a knowledge-based learning method that allows prior domain knowledge to be effectively utilized by machine learning systems. The domain knowledge is incorporated not into the learning algorithm itself but instead affects only the training data. The domain knowledge is used to explain and then transform the actual training examples into a more informative set of imaginary, or phantom examples. These phantom examples are added to the training set; the experienced examples are discarded. A new control policy is induced from the phantom training set. This policy is then exercised, yielding additional training points, and the process repeats.We investigate the performance of this method in a stylized air-hockey domain which demands a difficult nonlinear control policy. Our experiments show that, surprisingly, an accurate policy can be learned even if the domain theory is only imprecise and approximate. We advance an interpretation which indicates that the information available from a plausible qualitative domain theory is sufficient for robust successful learning. This interpretation is used to make a number of predictions which are tested in subsequent experiments. The outcomes confirm the interpretation and the robustness of the approach. 相似文献
998.
Distributed Control for 3D Metamorphosis 总被引:1,自引:1,他引:0
In this paper, we define Proteo as a class of three-dimensional (3D) metamorphic robotic system capable of approximating arbitrary 3D shapes by utilizing repeated modules. Each Proteo module contains embedded sensors, actuators and a controller, and each resides in a 3D grid space. A module can move itself to one of its open neighbor sites under certain motion constraints. Distributed control for the self-reconfiguration of such robots is an interesting and challenging problem. We present a class of distributed control algorithms for the reconfiguration of Proteo robots based on the goal-ordering mechanism. Performance results are shown for experiments of these algorithms in a simulation environment, and the properties of these algorithms are analyzed. 相似文献
999.
1000.
Umer Farooq Patricia Schank Alexandra Harris Judith Fusco Mark Schlager 《Computer Supported Cooperative Work (CSCW)》2007,16(4-5):397-429
Community computing has recently grown to become a major research area in human–computer interaction. One of the objectives
of community computing is to support computer supported cooperative work among distributed collaborators working toward shared
professional goals in online communities of practice. A core issue in designing and developing community computing infrastructures
– the underlying socio-technical layer that supports communitarian activities – is sustainability. Many community computing
initiatives fail because the underlying infrastructure does not meet end user requirements; the community is unable to maintain
a critical mass of users consistently over time; it generates insufficient social capital to support significant contributions
by members of the community; or, as typically happens with funded initiatives, financial and human capital resource become
unavailable to further maintain the infrastructure. Based on more than nine years of design experience with Tapped In – an
online community of practice for education professionals – we present a case study that discusses four design interventions
that have sustained the Tapped In infrastructure and its community to date. These interventions represent broader design strategies
for developing online environments for professional communities of practice. 相似文献