Evidence for the involvement of tyrosine-69 in the control of stereospecificity of porcine pancreatic phospholipase A2

We have studied the role of Tyr-69 of porcine pancreatic phospholipaseA₂ in catalysis and substrate binding, using site-directed mutagenesis.A mutant was constructed containing Phe at position 69. Kineticcharacterization revealed that the Phe-69 mutant has retainedenzymatic activity on monomeric and micellar substrates, andthat the mutation has only minor effects on k_cat and K_m. Thisshows that Tyr-69 plays no role in the true catalytic eventsduring substrate hydrolysis. In contrast, the mutation has aprofound influence on the stereospecificity of the enzyme. Whereasthe wild-type phospholipase A₂ is only able to catalyse thedegradation of sn-3 phospholipids, the Phe-69 mutant hydrolysesboth the sn-3 isomers and, at a low (1–2%) rate, the sn-1isomers. Despite the fact that the stereospecificity of themutant phospholipase has been altered, Phe-69 phospholipasestill requires Ca²⁺ ions as a cofactor and also retains itsspecificity for the sn-2 ester bond. Our data suggest that inporcine pancreatic phospholipase A₂ the hydroxyl group of Tyr-69serves to fix and orient the phosphate group of phospholipidmonomers by hydrogen bonding. Because no such interaction canoccur between the Phe-69 side-chain and the phosphate moietyof the substrate monomer, the mutant enzyme loses part of itsstereospecificity but not its positional specificity.     

Characterization of the isothermal crystallization of poly(ethylene terephthalate) by microhardness measurements

The two-stage isomerization isotherms of poly(ethylene Terephthalate) have been followed through microhardness analysis. A great variation in microhardness values seems to prove the increase of the strength and stiffness of the samples produced by the secondary isomerization. A smaller variation is observed for the primary isomerization.     

Synthesis,Structural Analysis,and Biological Evaluation of Thioxoquinazoline Derivatives as Phosphodiesterase 7 Inhibitors

PDE7 inhibitors regulate pro‐inflammatory and immune T‐cell functions, and are a potentially novel class of drugs particularly useful for treatment of a wide variety of immune and inflammatory disorders. Structural optimization of thioxoquinazoline derivatives led to new compounds with very interesting profiles as PDE7 or PDE7/PDE4 dual inhibitors, which may be further developed as new drugs for inflammatory and neurological diseases.

     

Iron Oxide Nanoparticles Employed as Seeds for the Induction of Microcrystalline Diamond Synthesis

Iron nanoparticles were employed to induce the synthesis of diamond on molybdenum, silicon, and quartz substrates. Diamond films were grown using conventional conditions for diamond synthesis by hot filament chemical vapor deposition, except that dispersed iron oxide nanoparticles replaced the seeding. X-ray diffraction, visible, and ultraviolet Raman Spectroscopy, energy-filtered transmission electron microscopy , electron energy-loss spectroscopy, and X-ray photoelectron spectroscopy (XPS) were employed to study the carbon bonding nature of the films and to analyze the carbon clustering around the seed nanoparticles leading to diamond synthesis. The results indicate that iron oxide nanoparticles lose the O atoms, becoming thus active C traps that induce the formation of a dense region of trigonally and tetrahedrally bonded carbon around them with the ensuing precipitation of diamond-type bonds that develop into microcrystalline diamond films under chemical vapor deposition conditions. This approach to diamond induction can be combined with dip pen nanolithography for the selective deposition of diamond and diamond patterning while avoiding surface damage associated to diamond-seeding methods.     

Study of the performance of Mo2C for iso-octane steam reforming

In the present work, the performance of commercial molybdenum carbide (Mo₂C) for isooctane steam reforming has been investigated in order to determine the effects of major operating parameters (temperature, space velocity, and steam to carbon ratio) on the catalytic activity. While the results obtained indicate an onset reforming temperature of 850 °C, high concentrations of H₂ in the reforming environment were found to reduce the onset temperature to 750 °C. The catalytic activity at 850 °C was sufficient to produce hydrogen yields greater than 90% and carbon conversions close to 100%, with a low selectivity to CH₄ and CO₂. In addition, and consistent with thermodynamic predictions, a steam to carbon ratio of 1 appeared to optimize the reforming rates. Finally, based on experimental observations, a reaction mechanism was formulated and used to interpret the results obtained during catalytic activity measurements. This mechanism involves continuous oxidation and reduction of Mo metal, which can provide activity and stability to the catalyst when occurring at similar rates.     

Synthesis of ω‐amino‐α‐phenylcarbonate alkanes and their polymerization to [n]‐polyurethanes

Aliphatic [n]‐polyurethanes have recently been synthesized from ω‐isocyanato‐α‐alkanols or, more traditionally, by cationic ring‐opening polymerization of cyclourethanes or by the Bu₂Sn(OMe)₂‐promoted polycondensation of ω‐hydroxy‐α‐O‐phenylurethane alkanes. For the latter procedures, the conditions employed do not seem to be suitable for highly functionalized monomers. In contrast, the polymerization of ω‐amino‐α‐phenylcarbonate alkanes is expected to occur under milder conditions. ω‐Amino‐α‐phenylcarbonate alkanes have been synthesized from 6‐aminohexanol (1) and 3‐aminopropanol (6). The procedure involves the N‐Boc protection of the amino group, followed by activation of the alcohol. Removal of the N‐Boc affords the corresponding ω‐amino‐1‐O‐phenyloxycarbonyloxyalkane hydrochlorides. Other oligomeric comonomers between 1 and 6 have been prepared. The polymerization of these precursors takes place in the absence of metal catalysts to afford the corresponding linear and regioregular [n]‐polyurethanes. The procedure described is useful for the preparation of stable ω‐amino‐α‐phenylcarbonate alkane derivatives, which possess varied chain lengths between the terminal functions. These monomers yield [n]‐polyurethanes having various structures starting from just two aminoalkanols. The polyurethanes were obtained in high yields, with reasonable molecular weight and polydispersity values, and they were characterized spectroscopically and thermally. These studies reveal constitutionally uniform structures that are free of carbonate or urea linkages. Copyright © 2010 Society of Chemical Industry     

Substrate effects on the mechanical properties and contact damage of diamond-like carbon thin films

Substrate effects on the mechanical properties and contact damage of diamond-like carbon (DLC) films have been investigated. To that end, a DLC film of 1 μm thickness was deposited on two different substrates: soda–lime glass (compliant and soft), and single-crystal silicon (stiff and hard). The elastic modulus and hardness were measured by means of nanoindentation. Quasi-static and sliding contact configurations were simulated by means of ultra-micro indentation, and surface and cross-sectional damage were assessed using a focused ion beam (FIB) miller. It was found that a compliant and soft substrate enhances crack initiation on the film surface in the form of ring/cone cracks. On the other hand, a stiff and hard substrate delays crack initiation in the film, but is more susceptible to fracture in the form of median and lateral cracks due to increased brittleness. The results have implications for the reliability of DLC-coated systems.     

Pore formation during continuous wave Nd:YAG laser welding of aluminium for automotive applications

Summary

Pore formation is an important concern in laser welding of automotive aluminium alloys. This paper investigates the influence of laser beam defocus on pore formation during continuous wave Nd:YAG laser welding of 5182 and 5754 automotive aluminium alloys. It was found that the instability of the keyhole during welding was a dominant cause of pore formation while hydrogen rejection played an insignificant role. The defocusing of the laser beam greatly affected the stability of the keyhole. Finally, the mechanism of keyhole collapse and pore formation is proposed.     

Self-feed device behaviour evaluation designed for assessment of operability of coated electrodes

The objective of the present work is to evaluate the effectiveness of a self-feed device for coated electrodes. This device is able to eliminate disturbances from manual operation or the use of automatic control systems during the study of coated electrode behaviour. The conjugation of different methods such as short-circuit time statistical analysis and metallographic analysis (penetration, heat-affected zone, shape and mean distance between successive solidification fronts) together with the analysis of the electric arc and electrode melting parameters allows us to obtain relationships between arc behaviour, mass transfer processes and bead appearance. This permits the extraction of criteria to describe process stability and the behaviour of the feed device employed. An appropriate statistical methodology was obtained to process the ‘duration of the short-circuit’ parameter through an appropriate adjustment of empirical distributions to a log-normal model. The self-feed device for coated electrode welding allows the repeatable and reproducible execution of welding beads in the flat position, keeping the arc length based on physical and chemical characteristics of the electrode during welding trials using 125, 140 and 160 A.