Single amino acid substitutions can further increase the stability of a thermophilic L-lactate dehydrogenase

Lactate dehydrogenases are of considerable interest as stereospecificcatalysts in the chemical preparation of enantiomerically pure-hydroxyacid synthons. For such applications in synthetic organicchemistry it would be desirable to have enzymes which tolerateelevated temperatures for prolonged reaction times, to increaseproductivity and to extend then applicability to poor substrates.Here, two examples are reported of significant thermostabilizations,induced by sitedirected mutagenesis, of an already thermostableprotein, the L-lactate dehydrogenase (EC 1.1.1.27 [EC] , 35 kDa permonomer subunit) from Bacillus stearothermophilus. Thermal inactivationof this enzyme is accompanied by irreversible unfolding of thenative protein structure. The replacement of Argl71 by Tyr stabilizesthe enzyme against thermal inactivation and unfolding. Thisstabilizing effect appears to be based on improved interactionsbetween the subunits in the core of the active dimeric or tetramericforms of the enzyme. The thermal stability of L-lactate dehydrogenasevariants with an active site Arg residue, either in the 171(wild-type) or in the 102 position, is further increased bysulfate ions. The two stabilizing effects are additive, as foundfor the Argl71Tyr/ Gln1O2Arg double mutant, for which the stabilityof the protein in 100 mM sulfate solution reaches that of L-lactatedehydrogenases from extreme thermophiles. All mutant proteinsretain significant catalytic activity, both in the presenceand absence of stnhilfoing salts, and are viable catalysts inpreparative scale reactions.     

Radical styrene polymerization in the presence of trace levels of sulfonic acids

The bulk polymerization of styrene in the presence of the vinyl functional sulfonic acid 2‐sulfoethylmethacrylate (SEM) was found to have utility for making polystyrenes with narrow polydispersity, bimodal polydispersity, and ultrahigh molecular weight at fast polymerization rates. Narrow polydispersity polymers were made by the addition of SEM to nitroxide‐mediated polymerizations. Bimodal polydispersity polymers were made by the ultrahigh molecular weight component being made in the presence of SEM in the absence of an initiator and the low molecular weight component being made in the presence of an initiator and/or chain‐transfer agent. Ultrahigh molecular weight monomodal polystyrenes were prepared at much faster polymerization rates than possible via spontaneous polymerization in the absence of SEM. SEM was found to be more effective, by an order of magnitude, than camphor sulfonic acid on a weight basis and, because it is copolymerized into the polymer chain, should not lead to corrosion problems during fabrication of the polymer. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 869–875, 2003     

3D-Printed Poly(acrylic acid–vinylimidazole) Ionic Polymer Metal Composite Actuators

Ionic polymer–metal composites (IPMC)—constructed using an ionic polymer sandwiched between metal electrodes—have shown great potential for the fabrication of soft actuators. IPMC architectures have many advantages including low actuation voltage, fast response, basic control, and relatively light weight. Poly(acrylic acid) (PAA)-based ion exchange membranes are of particular interest for IPMC devices due to their large ion exchange capacity and ease of preparation; however, they suffer from relatively weak mechanical strength. Here, PAA-based soft actuators are synthesized with enhanced mechanical properties and proton conductivity through the incorporation of hydrogen bonding interactions with imidazolium groups via copolymerization with 1-vinylimidazole. In addition to examining the impact of composition on physiochemical (swelling, glass transition, decomposition, Young's modulus, etc.) and electrochemical (specific capacitance) properties, an additive manufacturing process, digital light projection (DLP), is utilized to fabricate complex geometries demonstrating the potential for the fabrication of IPMC devices with complex actuation modalities. Planar DLP 3D-printed IPMC actuators of varied polymer compositions are fabricated with activated carbon and copper electrodes, and their actuation performance is evaluated in air, where large bending deformation is observed (14°–37°).     

Comparison of flocculated and dispersed single‐wall carbon nanotube‐based coatings using nonionic surfactants

Single‐wall carbon nanotubes (SWNTs) are promising filler materials for advanced polymer composites, but the impressive properties that have been predicted theoretically have not been realized experimentally. This gap is generally attributed to aggregation and nonideal dispersion of the SWNTs. Here, nonionic surfactants based on poly(ethylene oxide) are used to disperse SWNTs in either water or ethanol using sonication. The dispersed aqueous SWNTs are stable, while the analogous ethanol system yields loosely flocculated SWNTs. After drying these dispersions, the electrical conductivity of the flocculated system is at least an order of magnitude greater than the dispersed system at the same SWNT loading with conductivity greater than 20 S/cm obtained for the flocculated systems containing unsorted, commercial SWNTs. These flocculated systems can be readily sprayed to create conductive coatings. Despite their high electrical conductivity, these coatings provide only modest electromagnetic interference shielding (<20 dB) when testing large areas (30.5 × 30.5 cm²), which suggests significant heterogeneity or defects in these coatings that are not readily visible by eye or scanning electron microscopy. This defect mechanism is consistent with a decrease in shield efficacy at high SWNT loadings, despite no statistical change in the electrical conductivity of the coating. POLYM. ENG. SCI., 2013. © 2012 Society of Plastics Engineers     

Oxidation Transitions for SiC Part II. Passive‐to‐Active Transitions

Oxidation of SiC can occur in a passive mode, where a protective film is generated, or in an active mode, where a volatile suboxide is generated. The transitions from active‐to‐passive and passive‐to‐active are particularly important to understand as they occur via different mechanisms. In Part II of this article, the passive‐to‐active transition is explored. Three different types of SiC are examined—Si‐rich SiC, stoichiometric SiC, and C‐rich SiC. In addition to an in situ transition from passive‐to‐active, the effect of a preformed film on all three types of SiC is explored. It appears that the passive‐to‐active transition occurs when the SiO₂ scale begins to react with the SiC substrate. This reaction generates SiO(g) and CO(g), which build pressure beneath the SiO₂ scale, eventually causing the oxide to rupture. In addition, the SiO(g) can oxidize a distance away from the surface leading to the formation of SiO₂ needles and further promoting this SiO₂/SiC reaction. Thermodynamic and kinetic data are used to predict transition pressures of oxygen, which show reasonable agreement with those measured.     

Gene Expression Analyses Support Fallopian Tube Epithelium as the Cell of Origin of Epithelial Ovarian Cancer

Folate receptor alpha (FOLR1/FRA) is reported to be overexpressed in epithelial ovarian cancers (EOC), especially the serous histotype. Further, while dysregulation of the folate-dependent 1-carbon cycle has been implicated in tumorogenesis, little is known relative to the potential mechanism of action of FOLR1 expression in these processes. We therefore investigated the expression of FOLR1, other folate receptors, and genes within the 1-carbon cycle in samples of EOC, normal ovary and fallopian tube on a custom TaqMan Low Density Array. Also included on this array were known markers of EOC such as MSLN, MUC16 and HE4. While few differences were observed in the expression profiles of genes in the 1-carbon cycle, genes previously considered to be overexpressed in EOC (e.g., FOLR1, MSLN, MUC16 and HE4) showed significantly increased expression when comparing EOC to normal ovary. However, when the comparator was changed to normal fallopian tube, these differences were abolished, supporting the hypothesis that EOC derives from fallopian fimbriae and, further, that markers previously considered to be upregulated or overexpressed in EOC are most likely not of ovarian origin, but fallopian in derivation. Our findings therefore support the hypothesis that the cell of origin of EOC is tubal epithelium.     

Moisture absorption into ultrathin hydrophilic polymer films on different substrate surfaces

Moisture absorption into ultrathin poly(vinyl pyrrolidone) (PVP) films with varying thickness was examined using X-ray reflectivity (XR) and quartz crystal microbalance (QCM) measurements. Two different surfaces were used for the substrate: a hydrophilic silicon oxide (SiO_x) and a hydrophobic hexamethyldisilazane (HMDS) treated silicon oxide surface. The total equilibrium moisture absorption (solubility) was insensitive to the surface treatment in the thickest films (≈150 nm). However, strong reductions in the equilibrium uptake with decreasing PVP film thickness were observed on the HMDS surfaces, while the SiO_x surface exhibited thickness independent equilibrium absorption. The decreased absorption with decreasing film thickness is attributed a depletion layer of water near the polymer/HMDS interface, arising from hydrophobic interactions between the surface and water. The diffusivity of water decreased when the film thickness was less than 60 nm, independent of the surface treatment. Changes in the properties of ultrathin polymer films occur even in plasticized films containing nearly 50% water.