Multimer formation as a consequence of separate homodimerization domains: the human c-Jun leucine zipper is a transplantable dimerization module

Human c-Jun and c-Fos leucine zipper domains were examined fortheir ability to serve as autonomous dimerization domains aspart of a heterologous protein construct. Schistosoma japonicumglutathione S-transferase (GST) was fused to recombinant Junleucine zipper (rJunLZ) and Fos leucine zipper (rFosLZ) domains.SDS–PAGE ‘snapshot’ analyses based on disulphidelinkage of monomers demonstrated the ability of rJunLZ to functionas a dimerization motif in a foreign protein environment. Sterichindrance prevented formation of rJunLZ–GST::rFosLZ–GSTheterodimers whereas rJunLZ–GST::rFosLZ and rJunLZ::rFosLZ–GSTformed readily. Furthermore, rJunLZ–GST generated homodimerssuggesting fusion protein heterodimers interact differentlyto homodimers. Gel filtration chromatography confirmed thatGST is a dimer in solution and that attachment of a leucinezipper domain allows further interactions to take place. Sedimentationequilibrium analyses showed that GST is a stable dimer (K_a >10⁶ M^-1) with no higher multimeric forms. rFosLZ–GST weaklyassociates beyond a dimer (K_a {small tilde}4x10⁵ M^-1) and rJunLZ–GSTassociates indefinitely (K_a {small tilde}4x10⁶ M^-1), consistentwith an isodesmic model of association. The interaction of theseleucine zippers independently of GST association demonstratestheir utility in the modification of proteins when multimerformation is desired.     

Electric cadavers, 'metiphor,' and other medical software marvels

"Grateful Med," "Fluids," "Metiphor," virtual reality, and electric cadavers are just some of the programs and buzzwords in the futuristic world of medical software affecting many areas of health care.     

Business process management with the user requirements notation

A number of recent initiatives in both academia and industry have sought to achieve improvements in e-businesses through the utilization of Business Process Management (BPM) methodologies and tools. However there are still some inadequacies that need to be addressed when it comes to achieving alignment between business goals and business processes. The User Requirements Notation (URN), recently standardized by ITU-T, has some unique features and capabilities beyond what is available in other notations that can help address alignment issues. In this paper, a URN-based framework and its supporting toolset are introduced which provide business process monitoring and performance management capabilities integrated across the BPM lifecycle. The framework extends the URN notation with Key Performance Indicators (KPIs) and other concepts to measure and align processes and goals. An example process for controlling access to a healthcare data warehouse is used to illustrate and evaluate the framework. Early results indicate the feasibility of the approach.     

Determinants of the inhibitory action of purified 14-kDa phospholipases A2 on cell-free prothrombinase complex

It has been suggested (Kini, R. R., and Evans, H. J. (1987) J. Biol. Chem. 262, 14402-14407) that the anticoagulant activity of members of the 14-kDa phospholipase A2 (PLA2) family depends on the presence of basic residues within a variable surface region (residues 54-77) distinct from both the conserved catalytic machinery and surface sites mediating the antibacterial action of these enzymes (see Weiss, J., Inada, M., Elsbach, P., and Crowl, R. M. (1994) J. Biol. Chem. 269, 26331-26337). To further define the determinants of the anticoagulant activity of PLA2, we have analyzed the inhibitory effects of purified native and recombinant PLA2 on cell-free prothrombinase. Both native and recombinant wild-type pig pancreas (net charge -1) and human "secretory" PLA2 (net charge +15) produced similar dose-dependent inhibition of prothrombinase activity that was significantly less potent than a toxic PLA2 purified from snake venom. Site-specific mutations that either increased or decreased PLA2 activity toward bactericidal/permeability-increasing protein-treated Escherichia coli by up to 50-fold had no effect on antiprothrombinase activity. In contrast, substitution of Arg for Asp-59/Gly for Ser-60 in the pig PLA2 increased antiprothrombinase activity by 5-10-fold without affecting catalytic activity toward a range of phospholipid substrates or antibacterial activity. Comparison of antiprothrombinase activity of catalytically active and inactive forms of the PLA2 and under a range of phospholipid conditions revealed that the potent antiprothrombinase activity of native toxic venom PLA2 and of the D59R.S60G mutant pancreatic PLA2 reflect combined catalytic and noncatalytic actions, the latter apparently dependent on basic residues at discrete surface sites in the enzyme.     

Evolution of elastic modulus in roll forming

Roll forming is a continuous process in which a flat strip is incrementally bent to a desired profile. This process is increasingly used in automotive industry to form High Strength Steel (HSS) and Advanced High Strength Steel (AHSS) for structural components. Because of the large variety of applications of roll forming in the industry, Finite Element Analysis (FEA) is increasingly employed for roll forming process design. Formability and springback are two major concerns in the roll forming AHSS materials. Previous studies have shown that the elastic modulus (Young’s modulus) of AHSS materials can change when the material undergoes plastic deformation and the main goal of this study is to investigate the effect of a change in elastic modulus during forming on springback in roll forming. FEA has been applied for the roll forming simulation of a V-section using material data determined by experimental loading-unloading tests performed on mild, XF400, and DP780 steel. The results show that the reduction of the elastic modulus with pre-strain significantly influences springback in the roll forming of high strength steel while its effect is less when a softer steel is formed.     

Absorptivity contrast in transillumination imaging of tissue abnormalities

We calculate the time-resolved flux of photons transmitted across an optically turbid slab containing a partially absorbing inclusion. An analytical expression is obtained for the flux at a detector positioned opposite a point source (at a distance equal to the thickness of the slab) when the center of the inclusion lies on the line connecting those points. The calculation employs a discrete-time lattice random-walk model of photon transport. The resulting expression is used to assess the affects of time resolution on the detectability of the inclusion.