Conformational stability of factor VIIa: biophysical studies of thermal and guanidine hydrochloride-induced denaturation

The binding of the multidomain protein factor VIIa (fVIIa) to tissue factor provides the interprotein communication necessary to make fVIIa an efficient catalyst of the initial event in the extrinsic pathway of blood coagulation. We have investigated the stability of individual domains in fVIIa and the influence of Ca2+ and an irreversible active-site inhibitor (FFR-chloromethyl ketone). Equilibrium guanidine hydrochloride (GuHCl)-induced unfolding monitored by tryptophan fluorescence and far-UV circular dichroism (CD) demonstrated that the gamma-carboxyglutamic acid (Gla) domain unfolds at 0.3 M GuHCl and the serine protease (SP) domain at 3 M GuHCl and that Ca2+ is a prerequisite for the formation of an ordered, compact structure in the Gla domain. The loss of amidolytic activity coincides with the first transition, which is stabilized by the active-site inhibitor, and a change in the environment of the active site is demonstrated using a fluorescent inhibitor (DEGR-chloromethyl ketone). Thermal unfolding monitored by differential scanning calorimetry (DSC) reveals that Ca2+ stabilizes the SP domain slightly, increasing the unfolding temperature by 2.7 degrees C. In addition, Ca2+ is required for a large enthalpy change concomitant with unfolding of the Gla domain, and this unfolding enthalpy is only detectable in the presence of the SP domain, indicating some kind of interaction between these domains. Thermal unfolding measured by CD indicates secondary structural changes at the same temperature as the heat absorption in the DSC but only when both the Gla domain and the SP domain are present together with Ca2+ ions. Taken together, these results indicate a Ca2+-dependent interaction between the Gla domain and the SP domain, implying a high degree of flexibility of the domains in free fVIIa. It is also shown that the epidermal growth factor-like domains are stable at elevated temperatures and high GuHCl concentrations. Moreover, already at physiological temperature, subtle structural changes take place which influence the overall shape of fVIIa and are detrimental to its enzymatic activity.     

CTLA4 mediates antigen-specific apoptosis of human T cells

The regulation of T cell-mediated immune responses requires a balance between amplification and generation of effector function and subsequent selective termination by clonal deletion. Although apoptosis of previously activated T cells can be induced by signaling of the tumor necrosis factor receptor family, these molecules do not appear to regulate T-cell clonal deletion in an antigen-specific fashion. We demonstrate that cross-linking of the inducible T-cell surface molecule CTLA4 can mediate apoptosis of previously activated human T lymphocytes. This function appears to be antigen-restricted, since a concomitant signal T-cell receptor signal is required. Regulation of this pathway may provide a novel therapeutic strategy to delete antigen-specific activated T cells.     

Synthesis and characterization of block copolymers from D,L-lactide and poly(tetramethylene ether glycol)

The copolymer from D ,L -lactide and poly(tetramethyene ether glycol) (PTMG) was prepared in bulk with an isotributyl aluminum–water–phosphoric acid complex catalyst as the initiator and characterized by H-NMR, GPC, and DSC. The effects of the temperature and the amount of PTMG on the polymerization rate and the molecular weight of copolymers were studied. The behavior of the degradation and delivery rate of Levonorgestrel microspheres in vitro was observed. The results show that the degradation and the delivery rate can be controlled by adjusting the molar rate of hydrophilic and hydrophobic segments of the copolymer © 1995 John Wiley & Sons, Inc.     

Effect of hydrogen loading on temperature/electric-field poling ofSiO2-based thin films on Si

Waveguide stacks of GeSiO₂-based glass deposited on Si exhibit a permanent second-order nonlinearity following temperature electric-field poling. High temperature hydrogen loading increases the second harmonic signal under certain conditions     

Generating topological structures for surface models

Rapid prototyping has imposed new requirements on the processing of CAD models. To manufacture an object directly, the model must correspond to a real object and thus represent a closed volume. Its boundary must be continuous and closed, and no cracks or improper intersections are allowed. CAD models cannot always meet these requirements, especially when they are constructed by a surface modeler. The algorithm discussed analyzes surface models and generates a topological structure for them that meets rapid prototyping requirements for a closed volume with a continuous, closed boundary     

Grassmannian, central projection, and output feedback poleassignment of linear systems

A sufficient rank condition for arbitrary pole assignability by static output feedback of m-input, p-output linear systems of McMillan degree n相似文献   

Invasiveness of LiTaO/sub 3/ and GaAs probes in external E-O sampling

Disturbances of signals on a coplanar waveguide (CPW) induced by the presence of LiTaO/sub 3/ and GaAs electrooptic probes in external electrooptic (EO) sampling have been simulated and compared quantitatively. The finite-difference-time-domain method is used to simulate the full wave field around a coplanar waveguide on a GaAs substrate in an external EO sampling configuration. The results indicate that the induced signal disturbance, or invasiveness, of a LiTaO/sub 3/ probe is almost ten times that of a GaAs probe in terms of the magnitude of S/sub 11/, but that LiTaO/sub 3/ yields about two times the EO response for a given S/sub 11/ and optical probing wavelength. The transparency of LiTaO/sub 3/ to shorter wavelengths, however, allows an even higher sensitivity for this material relative to GaAs. The results suggest that these probes do not exhibit significant invasiveness (magnitude of S/sub 11/ smaller than -40 dB), if they are removed from contact by the distance of CPWs center conductor width.<>     

Effect of silver addition on superconductivity in the Bi1.6Pb0.4Sr1.6Ca2Cu3O10−y system

Silver/superconductor composites containing 0 to 80 vol% silver have been prepared and their properties determined. Optimum heat treatment at sintering temperatures ( 800° C) under low oxygen pressures produces material with high critical current density and improves physical properties. Magnetic susceptibility measurements have been found to be consistent with resistivity results. In order to retain a single high-T _c phase with increasing silver content, decreased oxygen partial pressures are required. Using the normal-state resistivity of these composites, a percolation threshold at a silver volume fraction of 43% was observed, while zero resistivity measurements show that a continuous superconducting network can be obtained with up to 80 vol% silver. The critical current density of 21 vol% silver-doped samples was found to be 1520 A cm^–2 at 77.3 K, compared to 260 A cm^–2 for an undoped sample.     

Scaling effects on flow hydrodynamics of confined microdroplets induced by Rayleigh surface acoustic wave

This paper reports an experimental and numerical investigation on the scaling effects in the flow hydrodynamics for confined microdroplets induced by a surface acoustic wave (SAW). The characteristic parameters of the flow hydrodynamics were studied as a function of the separation height, H, between the LiNbO₃ substrate and a top glass plate, for various droplets volumes and radio-frequency powers. The ratio of the gap height to attenuation length of the SAW, H/l _SAW, is shown to be an important parameter affecting the streaming flow induced in this confined regime. The reported numerical and experimental results are in good agreement over the range examined in this study and demonstrate that, at a lower gap heights of H?≤?100?μm, a significant decrease in streaming velocity or Reynolds number is induced, with the velocity approaching zero when the gap height is decreased to ~50?μm. An increase in the gap height results in an increased streaming velocity; however, if the gap height exceeds 70?% of the SAW attenuation length, any further increase in the gap height induces a drop in the streaming velocity.     

Laser glazing of an Fe-C-Sn alloy

Morphology and geometry of melted zones, cooling rates, microstructure and microhardness in the laser-glazed Fe-4%C-10%Sn alloy have been investigated. The computer simulation on the basis of the moving gaussian source model was used successfully to predict the maximum width and depth of the melted zone and the cooling rate. The microstructure from the surface to the bottom of the laser-melted zone is a non-crystalline phase, dendritic grains and a microcrystalline zone successively. Values of the averaged-spacing of the non-crystalline phase are 0.205⁶ and 0.121⁹nm, respectively; twinned martensites, having an axial ratioc/a of 1.128, existed in dendritic grains, and carbides of Fe₃ C at the interdendritic regions; the microcrystalline zone was composed of -Fe and a new bet (a=0.415 nm,c=0.955 nm) phase. The different microstructure in the melted zone can be explained by the results of the heat flow calculation. A fine eutectic structure (-Fe + Fe₃C) was observed in heat-affected zones. Microhardness of the eutectic structure can be predicted by the empirical relation of fracture stress to the interlamellar spacing of pearlite.