Biochemical studies of the mechanism of action of the Cdc42-GTPase-activating protein

The small GTP-binding proteins Rac, Rho, and Cdc42 were shown to mediate a variety of signaling pathways including cytoskeletal rearrangements, cell-cycle progression, and transformation. Key to the proper function of these GTP-binding proteins is an efficient shut-off mechanism that ensures the decay of the signal. Regulatory proteins termed GAPs (GTPase-activating proteins) enhance the intrinsic GTP hydrolysis of the GTP-binding proteins, thereby ensuring signal termination. We have used site-specific mutagenesis to elucidate the limit domain for GAP activity in Cdc42-GAP, and show that in addition to the known GAP-homology domain (three conserved boxes), a C-terminal region outside that domain is also essential for GAP activity. In addition, we have replaced the conserved arginine (Arg305), which was suggested by structural studies to be a key catalytic residue, with an alanine and found that the R305A Cdc42-GAP mutant has a greatly diminished catalytic capacity but is still able to bind Cdc42 with high affinity. Thus, a key catalytic role for this residue is confirmed. However, we also present evidence for the involvement of an additional residue(s), since the R305A Cdc42-GAP mutant still exhibits measurable activity. Some of this residual activity might result from a neighboring arginine, since a double mutant R305A/R306A shows a further decrease in catalytic activity.     

Inference rules using local contexts

We construct equivalent localized versions of a formula, adding assumptions simultaneously to various locations, where the particular location determines what is added. Inference rules that take advantage of localized formulas are presented for sequent calculi in which the left hand side of sequents can be used to accumulate the background assumptions (or contexts) of assertions. The intended application is to the automatic generation of tractable justifying lemmas for substitution operations for interactive proof development systems, especially those concerned with mathematical topics where manipulation of deeply embedded terms is desirable.     

Adsorption of dissolved organics from industrial effluents on to activated carbon

Industrial effluents usually include multicomponent organic solutes. The optimum pH for adsorption of a specific industrial effluent on activated carbon should be determined experimentally because, in general, more than one mechanism is involved. A series of experiments was conducted to establish the influence of the initial hydrogen ion concentration on carbon adsorption of organic solutes. For these studies, powdered activated carbon was used, and the water systems studied included both single component pure organic compounds as well as multicomponent organic wastes. Results indicate that the pH effect upon the effectiveness of carbon adsorption mainly depends upon the nature of the adsorbed substance. In general, the degree of ionisation is the controlling factor for adsorption of ionic organic solutes on activated carbon. Adsorption reaches a maximum at the point of least ionisation of the adsorbate. As the organic compounds become more complex (i.e. longer hydrocarbon chains, higher molecular weights, increased branching), the electrical adsorption forces between activated carbon and ionic organic solutes will govern. Anionic surfactants meet with decreased electronegative repulsive forces at low pH levels, which increase the effectiveness of carbon adsorption. However, the adsorption of a cationic surfactant is increased by an increase in the electronegative carbon surface at high pH levels. When ionic organic solutes become much more complex, like a polymer, the effects of both ionisation and electrical adsorption forces become less important. Instead, the adsorption rate will be controlled by the extent of hydrolysis caused by the pH adjustment. For non-ionic organic solutes, chemical reaction(s) between the adsorbate and the added chemical (acid or base) for pH adjustment is an important controlling factor. Again, hydrolysis is responsible for the breakdown of larger size molecules to smaller sizes. Then an increase in adsorption rate with decreasing molecular weight of adsorbate is expected.     

Silicon Nitride Derived from an Organometallic Polymeric Precursor: Preparation and Characterization

Partially crystalline Si₃N₄, with nanosized crystals and a specific surface area greater than 200 m²/g, is obtained by pyrolysis of a commercially available vinylic polysilane in a stream of anhydrous NH₃ to 1000°C. This polymer does not contain N initially. Crystallization to high-purity α-Si₃N₄ proceeds with additional heating above 1400°C under N₂. The changes in crystallinity, powder morphology, infrared spectra, and elemental compositions, for samples annealed from 1000° to 1600°C under N₂, are consistent with an amorphous-to-crystalline transformation. Although macroscopic consolidation and local densification occur at 1400°C, volatilization and accompanying weight loss limit bulk densification. The effect of temperature on specific surface area is examined and related to the sintering process. These results are applicable to pyrolysis, decomposition, and crystallization studies of ceramics synthesized by polymeric precursor routes.     

Preparation of Silicon Carbide/Aluminum Nitride Ceramics Using Organometallic Precursors

Solid solutions of 2H -SiC/AlN can be prepared at temperatures less than 1600°C by rapid pyrolysis ("hot drop") of mixtures of [(Me₃Si)_0.80((CH₂=CH)MeSi)_1.0(MeHSi)_0.35] _n (VPS) or [MeHSiCH₂] _n (MPCS) with [R₂AlNH₂]₃, where R=Et, i -Bu or simply by slow pyrolysis of the precursor mixture in the case of [Et₂AlNH₂]₃. In contrast, slow pyrolysis of mixtures of VPS or MPCS with [ i -Bu₂AlNH₂]₃ yields a composite of 2 H -AlN and 3 C -SiC at 1600°C, which transforms into a single 2 H -SiC/AlN solid solution on heating to 2000°C. The influences of the nature of the precursor and processing conditions on the structure, composition, and purity of the SiC/AlN materials are discussed.     

Preparation of lot samples of nut meats for mycotoxin assay

A procedure has been devised for preparing lot samples of mycotoxin-contaminated nut meats so that a representative analytical sample may be removed. The sample is rapidly reduced to coarse size. A relatively large portion (about 1/10 of total sample) of subsample is then split out and further comminuted to a fine particle size with the aid of a fat solvent (meat-solvent, w/v, 3:2). The analytical sample is removed from this mixture. The procedure was tested with shelled almonds and shelled walnuts using radioactive nuts to simulate the mycotoxin contamination and provide a simple, precise measure of the contaminated nut meat distribution. The pooled coefficient of variation was 18% for the subsamples and 4.4% for the analytical samples. Considering the dilution factors used (1.50 and 2.14 contaminated nuts/10⁴ nuts) and the low degree of reliability of the lot sample, the sample preparation methods tested appear to be practical and reliable.     

Chemical modification of hemp,sisal, jute,and kapok fibers by alkalization

Plant fibers are rich in cellulose and they are a cheap, easily renewable source of fibers with the potential for polymer reinforcement. The presence of surface impurities and the large amount of hydroxyl groups make plant fibers less attractive for reinforcement of polymeric materials. Hemp, sisal, jute, and kapok fibers were subjected to alkalization by using sodium hydroxide. The thermal characteristics, crystallinity index, reactivity, and surface morphology of untreated and chemically modified fibers have been studied using differential scanning calorimetry (DSC), X‐ray diffraction (WAXRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM), respectively. Following alkalization the DSC showed a rapid degradation of the cellulose between 0.8 and 8% NaOH, beyond which degradation was found to be marginal. There was a marginal drop in the crystallinity index of hemp fiber while sisal, jute, and kapok fibers showed a slight increase in crystallinity at caustic soda concentration of 0.8–30%. FTIR showed that kapok fiber was found to be the most reactive followed by jute, sisal, and then hemp fiber. SEM showed a relatively smooth surface for all the untreated fibers; however, after alkalization, all the fibers showed uneven surfaces. These results show that alkalization modifies plant fibers promoting the development of fiber–resin adhesion, which then will result in increased interfacial energy and, hence, improvement in the mechanical and thermal stability of the composites. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 2222–2234, 2002     

Equilibrium bulk polymerization of 1,3-dioxolan

Cationic bulk polymerization of 1,3-dioxolan has been carried out in sealed ampoules using a high vacuum technique. The polymerization is initiated with triethyl oxonium hexafluorophosphate and the equilibrium between monomer and active polymer is attained within a few hours. Specific volumes of pure monomer and polymer in solution of its own monomer have been measured. Equilibrium measurements have been performed in the 40° to 141·4°C temperature range and the ceiling temperature is estimated to be 144° ± 2°C. The effect of short polymer chains on the equilibrium is discussed briefly. Values of ΔG_lc, the free energy of polymerization of one mole of pure liquid monomer to one base-mole of amorphous polymer, are computed making allowance for the non-ideal mixing. Respective values of ?17.5 ± 0.8 kJ/mol and ?47.9 ± 2.2JK^?1mol^?1 are deduced for the corresponding ΔH_lc and ΔS_lc. ΔG_lc is also computed from published data on equilibrium polymerization of 1,3-dioxolan in various solvents and the combined results for both types of polymerization yield ΔH_lc = ?16.7 ± 0.5kJ/mol and ΔS_lc = ?45.8 ± 1.5JK^?1mol^?1 for the 20° to 140°C range.     

Factor Graph Modeling of Rigid‐body Dynamics for Localization,Mapping, and Parameter Estimation of a Spinning Object in Space

This paper presents a new approach for solving the simultaneous localization and mapping problem for inspecting an unknown and uncooperative object that is spinning about an arbitrary axis in space. This approach probabilistically models the six degree‐of‐freedom rigid‐body dynamics in a factor graph formulation. Using the incremental smoothing and mapping system, this method estimates a feature‐based map of the target object, as well as this object's position, orientation, linear velocity, angular velocity, center of mass, principal axes, and ratios of inertia. This solves an important problem for spacecraft proximity operations. Additionally, it provides a generic framework for incorporating rigid‐body dynamics that may be applied to a number of other terrestrial‐based applications. To evaluate this approach, the Synchronized Position Hold Engage Reorient Experimental Satellites (SPHERES) were used as a testbed within the microgravity environment of the International Space Station. The SPHERES satellites, using body‐mounted stereo cameras, captured a dataset of a target object that was spinning at ten rotations per minute about its unstable, intermediate axis. This dataset was used to experimentally evaluate the approach described in this paper, and it showed that it was able to estimate a geometric map and the position, orientation, linear and angular velocities, center of mass, and ratios of inertia of the target object.     

Considerations for linking seatpan and backrest angles

Modern ergonomic chairs typically have several dimensions that can be adjusted independently of one another. Finding a desirable setting for any one dimension can depend on how other dimensions are set, thereby confronting users with a significant control problem. One design strategy for dealing with this problem has been to link changes in seatpan and backrest angles in some ratio, such that a one‐degree change in seatpan angle is associated with a two‐ or three‐degree change in backrest angle. However, there is no evidence to justify the choice of a particular ratio. This article presents data that addresses this issue. Subjects, performing either an entry or verification task, could adjust the chair to any position. Backrest and seatpan angles were plotted over time and analyzed using both graphical and statistical methods. The resulting scatter plots do not support the industry standard, 1:2 or 1:3 ratio, of changes in seatpan to backrest angles. The possibility of a variable linkage is discussed; however, problems associated with such a solution raise the possibility that control issues might be best addressed through training and exploration.