Construction, properties and specific fluorescent labeling of a bovine prothrombin mutant engineered with a free C-terminal cysteine

To define the role of phosphatidylserine-induced conformationalchanges in prothrombin activation during blood coagulation,a recombinant bovine prothrombin was constructed, characterizedand shown to have a globally native-like conformation. We introduceda cysteine to replace the penultimate residue (Gly581) of apreviously constructed active site mutant, and expressed thedouble mutant in Chinese hamster ovary cells at the level of0.6 µg/ml of cell culture medium. Specific labeling withfluorescein maleimide was accomplished by limited reductionwith dithiothreitol to free the engineered cysteine while maintainingthe native-like functional properties of the molecule. The averagestoichiometry of labeling was 0.84 probe/protein. The locationof the probe at the C-terminus was confirmed by proteolysisby native thrombin, by Taipan venom, and by carboxypeptidaseY. Both the double mutant and labeled prothrombin could be activatedby snake venoms and the prothrombinase but, as expected, thedouble mutant meizothrombin did not autolyze as does nativemeizothrombin. Thus, for the first time, a native-like but specificallylabeled prothrombin has been constructed. This molecule willbe an essential tool for elucidating the structural role ofmembranes during prothrombin activation. In addition, the methodsdescribed might be usefully applied to labeling of an odd, engineeredcysteine in other disulfide bond-containing proteins.     

Reduction of sulphur in PbSPbCl2NaCl melts

The reduction of dissolved sulphur and the soluble species formed during the electrolysis of PbS dissolved in the PbCl₂NaCl eutectic has been studied chronopotentiometrically. The pre-lead reduction process reported earlier[1] can now be assigned to a subsulphide species that is generated electrochemically. At higher PbS concentrations (eg 10^?3 mol cm^?3), elemental sulphur introduces a new reduction wave in addition to the subsulphide. The saturation solubility of sulphur is approximately proportional to the square of the PbS concentration. It has been estimated to be 2.3× 10^?4 mol S cm^?3 at 430°C for a melt containing 2.4× 10^?3 mol PbS cm^?3 in the PbCl₂-NaCl eutectic.     

Public Displays of Play: Studying Online Games in Physical Settings

As research on virtual worlds gains increasing attention in educational, commercial, and military domains, a consideration of how player populations are ‘reassembled’ through social scientific data is a timely matter for communication scholars. This paper describes a large‐scale study of virtual worlds in which participants were recruited at public gaming events, as opposed to through online means, and explores the dynamic relationships between players and contexts of play that this approach makes visible. Challenging conventional approaches to quantitatively driven virtual worlds research, which categorizes players based on their involvement in an online game at a particular point in time, this account demonstrates how players' networked gaming activities are contingent on who they are playing with, where, and when.     

Polyurethane‐infiltrated carbon foams: A coupling of thermal and mechanical properties

The thermal and mechanical properties of polyurethane‐infiltrated carbon foam of various densities were investigated. By combining the high thermal conductivity of the carbon foam with the mechanical toughness of the pure polyurethane, a mechanically tough composite (relative to the unfilled foam) that could be used at higher temperatures than the polyurethane's degradation was formed. Both the tensile strength and the modulus increased by an order of magnitude for the composites compared to unfilled foam, while the compressive and shear strengths and moduli of the composites approached values exhibited by pure polyurethane. At both 300 and 400°C, the rectangular blocks of pure polyurethane lost their mechanical integrity due to decomposition in air. Thermogravimetric analysis confirms substantial initial weight loss above 290°C. Filled carbon foam blocks, however, maintain their mechanical integrity at both 300 and 400°C indefinitely, although the bulk of the rectangular block mass is polyurethane. Three different carbon foam densities are examined. As expected, the higher density foams show greater heat transfer. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 87: 2348–2355, 2003     

Transformation Toughening in Large-Grain-Size CeO2-Doped ZrO2 Polycrystals

The fracture and transformation behavior of tetragonal polycrystalline ZrO₂ alloys containing 18 mol% CeO₂ (Ce-TZP) was investigated. In the absence of applied stress the tetragonal phase was found to be stable in large-grained (>30 μm) samples at room temperature. The monoclinic phase was detected in regions of high residual stress near hardness indentations although no evidence of a wake of monoclinic phase along the fracture surface was observed. The fracture toughness increased from 4 to 7 MPa · m^1/2 as density and/or grain size increased. It is proposed that the relatively high toughness of these materials is due to the occurrence of stress-driven tetragonal-to-monoclinic transformation near the crack tip, which reverses when the crack has passed.     

Factors influencing the photofading of commercial anthraquinone dyes in solution

The photofading of two anthraquinone dyes has been studied in aqueous solution using ultraviolet-visible absorption spectroscopy and flash photolysis. The influence of alcohol concentration, pH, atmosphere, photosensitisers and stabilisers have been examined and these lead to some important conclusions on the mechanism of dye fading. Essentially, the results indicate the photoexcited triplet state of the dye undergoes a process of either electron or hydrogen-atom abstraction depending on the nature of the environment. Other factors such as aggregation and singlet oxygen also appear to play an important role in solution photofading. The relevance of these results to photofading in a polymeric phase is discussed.     

Microstructure and Mechanical Properties of Mullite–Silicon Carbide Composites

Mullite-SiC-whisker composites were prepared by powder processing using two commercial SiC whiskers. These composites were prepared by sintering rather than hot-pressing. A mulliteSlC-powder composite and a base line mallite material were also prepared for comparison with the two whisker composite materials. Fracture toughness measurements showed significant enhancement in only one of the whisker composite materials. The microstructure of the four materials was examined by scanning electron microscopy and transmission electron microscopy to assist in the explanation of the mechanical behavior of these composites. The examinations suggested that most of the toughening results from second-phase particles, with only limited toughening from effects associated with whiskers per se. In one case, higher toughness was partially associated with the formation of sialon phase by reaction with the whiskers and the furnace environment.     

Modeling the Value-Based Software Process with Object-Petri-Nets

Commercial organizations increasingly need software processes sensitive to busi- ness value, quick to apply, supportive of multi-stakeholder collaboration, and capable of early analysis for subprocess consistency and compatibility. This paper and a companion paper \Applying Value-Based Software Process: An ERP Example" present our experience in ap- plying a lightweight synthesis of a Value-Based Software Quality Achievement (VBSQA) process and an Object-Petri-Net (OPN) based process model to achieve a stakeholder win- win outcome for software quality achievement in an ERP software project in China. We attempt to answer such questions as (1) whether it is possible to model various project stakeholder perspectives using formal process modeling languages in a real-world project? (2) how to make stakeholders synchronize and stabilize their value propositions, activities and commitments as needed. The companion paper presents the VBSQA process and its application to the ERP project; this paper presents the OPN-based process modeling and its application. The application results of OPN process modeling conˉrmed that (1) the OPN- based process model provided project managers with a synchronization and stabilization framework for process activities, success-critical stakeholders and their value propositions; (2) process visualization and simulation tools signiˉcantly increased management visibility and controllability for the success of the software project.     

PID controller tuning for a multivariable glass furnace process by genetic algorithm

Standard genetic algorithms (SGAs) are investigated to optimise discrete-time proportional-integral-derivative (PID) controller parameters, by three tuning approaches, for a multivariable glass furnace process with loop interaction. Initially, standard genetic algorithms (SGAs) are used to identify control oriented models of the plant which are subsequently used for controller optimisation. An individual tuning approach without loop interaction is considered first to categorise the genetic operators, cost functions and improve searching boundaries to attain the desired performance criteria. The second tuning approach considers controller parameters optimisation with loop interaction and individual cost functions. While, the third tuning approach utilises a modified cost function which includes the total effect of both controlled variables, glass temperature and excess oxygen. This modified cost function is shown to exhibit improved control robustness and disturbance rejection under loop interaction.     

Reviewing VTAM and NetView Security Controls

Abstract

The basic premise of developing any information system is to increase efficiency and manage information in an improved manner. Based on the definitions above, an information system is “a group of devices or artificial objects serving a common purpose to communicate knowledge or intelligence.” Most information systems evolve from processes that either manipulate much information or rely on complex calculations. In other words, information systems are developed either to manage information or to manipulate it. Systems are meant to receive input, manipulate the input, and provide output.