The Photosensitising Clinical Agent Verteporfin Is an Inhibitor of SPAK and OSR1 Kinases

STE20/SPS1‐related proline/alanine‐rich kinase (SPAK) and oxidative‐stress‐responsive kinase 1 (OSR1) are two serine/threonine protein kinases that play key roles in regulating ion homeostasis. Various SPAK and OSR1 mouse models exhibited reduced blood pressure. Herein, the discovery of verteporfin, a photosensitising agent used in photodynamic therapy, as a potent inhibitor of SPAK and OSR1 kinases is reported. It is shown that verteporfin binds the kinase domains of SPAK and OSR1 and inhibits their catalytic activity in an adenosine triphosphate (ATP)‐independent manner. In cells, verteporfin was able to suppress the phosphorylation of the ion co‐transporter NKCC1; a downstream physiological substrate of SPAK and OSR1 kinases. Kinase panel screening indicated that verteporfin inhibited a further eight protein kinases more potently than that of SPAK and OSR1. Although verteporfin has largely been studied as a modifier of the Hippo signalling pathway, this work indicates that the WNK‐SPAK/OSR1 signalling cascade is also a target of this clinical agent. This finding could explain the fluctuation in blood pressure noted in patients and animals treated with this drug.     

Decoupled Roles for the Atypical,Bifurcated Binding Pocket of the ybfF Hydrolase

Serine hydrolases have diverse intracellular substrates, biological functions, and structural plasticity, and are thus important for biocatalyst design. Amongst serine hydrolases, the recently described ybfF enzyme family are promising novel biocatalysts with an unusual bifurcated substrate‐binding cleft and the ability to recognize commercially relevant substrates. We characterized in detail the substrate selectivity of a novel ybfF enzyme from Vibrio cholerae (Vc‐ybfF) by using a 21‐member library of fluorogenic ester substrates. We assigned the roles of the two substrate‐binding clefts in controlling the substrate selectivity and folded stability of Vc‐ybfF by comprehensive substitution analysis. The overall substrate preference of Vc‐ybfF was for short polar chains, but it retained significant activity with a range of cyclic and extended esters. This broad substrate specificity combined with the substitutional analysis demonstrates that the larger binding cleft controls the substrate specificity of Vc‐ybfF. Key selectivity residues (Tyr116, Arg120, Tyr209) are also located at the larger binding pocket and control the substrate specificity profile. In the structure of ybfF the narrower binding cleft contains water molecules prepositioned for hydrolysis, but based on substitution this cleft showed only minimal contribution to catalysis. Instead, the residues surrounding the narrow binding cleft and at the entrance to the binding pocket contributed significantly to the folded stability of Vc‐ybfF. The relative contributions of each cleft of the binding pocket to the catalytic activity and folded stability of Vc‐ybfF provide a valuable map for designing future biocatalysts based on the ybfF scaffold.     

Adhesion of a Micro Double Cantilever Beam Driven by Capillary Forces

Capillary forces present during the manufacturing and operation of microelectromechanical structures have been shown to cause collapse of these structures. Once collapsed these structures may permanently adhere together. In this paper we study the adhesion of a microstructure represented by a double cantilever beam (DCB); such adhesion is driven by both the capillary forces from a liquid droplet located between the DCB and the adhesion forces between the solids. The DCB is modeled using both a beam theory that accounts for geometrical nonlinearity and the linearized beam theory. This model rigorously couples DCB deformation, capillary forces, meniscus movement and adhesion, and reveals four dimensionless parameters that govern the adhesion problem. How changes in these parameters influence the capillary forces, the length of the contact zone, and the contact force is discussed in detail.     

Toughening of polylactide by melt blending with linear low‐density polyethylene

Melt blending of polylactide and linear low‐density polyethylene (LLDPE) was performed in an effort to toughen polylactide. In addition, two model polylactide‐polyethylene (PLLA‐PE) block copolymers were investigated as compatibilizers. The LLDPE particle size and the impact resistance of binary and ternary blends were measured to determine the extent of compatibilization. For the amorphous polylactide (PLA), toughening was achieved only when a PLLA‐PE block copolymer was used as a compatibilizer. For the semicrystalline polylactide (PLLA), toughening was achieved in the absence of block copolymer. To decrease the variability in the impact resistance of the PLLA/LLDPE binary blend, as little as 0.5 wt % of a PLLA–;PE block copolymer was effective. The differences that were seen between the PLA and PLLA binary blends were investigated with adhesion testing. The semicrystalline PLLA did show significantly better adhesion to the LLDPE. We propose that tacticty effects on the entanglement molecular weight or miscibility of polylactide allow for the improved adhesion between the PLLA and LLDPE. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 3757–3768, 2003     

Structure–property relationships in BaTiO3–BiFeO3–BiYbO3 ceramics

Ba_1?xBi_xTi_1?xYb_x/2Fe_x/2O₃ ceramics were fabricated by the solid state reaction method. X-ray diffraction analyses show 0 ≤ x ≤ 0.04 ceramics to have an average crystal structure described by the non-centrosymmetric tetragonal P4 mm space group, whereas x ≥ 0.08 ceramics are consistent with a centrosymmetric cubic perovskite (space group Pm-3 m). Coexistence of both tetragonal and cubic symmetries is observed for x = 0.06. Raman spectroscopy analysis corroborate a change in average structure with increasing x, but also show the local crystal symmetry for x ≥ 0.08 ceramics to deviate from the idealized cubic perovskite structure. Dielectric data show a ferroelectric-to-relaxor crossover, which occurs in conjunction with the change in both the average and local crystal symmetry as indicated by X-ray and Raman data. For x ≥ 0.08, ceramics exhibit relaxor behavior, which is also accompanied by a shift of the permittivity maxima towards higher temperatures with increasing x.     

Native state kinetic stabilization as a strategy to ameliorate protein misfolding diseases: a focus on the transthyretin amyloidoses

Small molecule-mediated protein stabilization inside or outside of the cell is a promising strategy to treat protein misfolding/misassembly diseases. Herein we focus on the transthyretin (TTR) amyloidoses and demonstrate that preferential ligand binding to and stabilization of the native state over the dissociative transition state raises the kinetic barrier of dissociation (rate-limiting for amyloidogenesis), slowing and in many cases preventing TTR amyloid fibril formation. Since T119M-TTR subunit incorporation into tetramers otherwise composed of disease-associated subunits also imparts kinetic stability on the tetramer and ameliorates amyloidosis in humans, it is likely that small molecule-mediated native state kinetic stabilization will also alleviate TTR amyloidoses.     

Determination of the vapor pressures of select polychlorinated dibenzo-p-dioxins and dibenzofurans at 75-275 °C

Vapor pressures were determined for several polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) at 75-275 °C, extending the available literature data to more relevant temperature regions and providing the first experimental data for 2,3,7-trichlorodibenzo-p-dioxin (2,3,7-TriCD). A modification of the effusion technique, based upon controlling the diffusion of the target compound and subsequent high resolution gas chromatography/low resolution mass spectrometry (HRGC/LRMS) analysis, was proven comparable to other accepted methods for determining the vapor pressures of semi-volatile organic compounds (SVOCs). Vapor pressures for octachlorodibenzo-p-dioxin (OCDD) and octachlorodibenzofuran (OCDF) were in excellent agreement with those reported in literature. The application of the current method for the vapor pressure determinations of eight polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) in the extended temperature range (up to 275 °C) is reported. The extension of the vapor pressures to such temperatures, unprecedented for the PCDDs/Fs, is important for vapor-particle partitioning modeling in regions relevant to PCDD/F formation and control. Estimates for the melting temperatures and enthalpies of sublimation and vaporization are also reported, the latter for which no experimentally determined values have been found in the literature. The use of the method to deliver reproducible, trace concentrations (ppt-ppb) of targets was applied to the calibration of the jet-REMPI/TOFMS as an online detector for low chlorinated PCDDs/Fs.     

Modeling and analysis of thickness gradient and variations in vacuum‐assisted resin transfer molding process

As vacuum‐assisted resin transfer molding (VARTM) is being increasingly used in aerospace applications, the thickness gradient and variation issues are gaining more attention. Typically, thickness gradient and variations result from the infusion pressure gradient during the process and material variations. Pressure gradient is the driving force for resin flow and the main source of thickness variation. After infusion, an amount of pressure gradient is frozen into the preform, which primarily contributes to the thickness variation. This study investigates the mechanism of the thickness variation dynamic change during the infusion and relaxing/curing processes. A numerical model was developed to track the thickness change of the bagging film free surface. A time‐dependent permeability model as a function of compaction pressure was incorporated into an existing resin transfer molding (RTM) code for obtaining the initial conditions for relaxing/curing process. Control volume (CV) and volume of fluid (VOF) methods were combined to solve the free surface problem. Experiments were conducted to verify the simulation results. The proposed model was illustrated with a relatively complex part. POLYM. COMPOS., 2008. © 2008 Society of Plastics Engineers