Microbial Conversion of Vegetable Oil to Rare Unsaturated Fatty Acids and Fatty Alcohols by an <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis> Isolate

We isolated two new microorganisms capable of converting vegetable oil to several rare unsaturated fatty acids and rare unsaturated fatty alcohols from a soil sample. The strains were identified as belonging to the same genus and species, Aeromonas hydrophila. The rare unsaturated fatty acids and rare unsaturated fatty alcohols were accumulated as a wax ester form by the strains. Compared to other strains, the A. hydrophila isolates effectively decreased fatty acid chain lengths and converted rapeseed oil, which is rich in 9-C18:1 fatty acid, into rare fatty acids, such as 7-C16:1 fatty acid and 5-C14:1 fatty acid. Furthermore, the A. hydrophila isolates converted the resulting fatty acids to rare unsaturated fatty alcohols, such as 7-C16:1 fatty alcohol and 5-C14:1 fatty alcohol. The isolates also converted safflower oil, which is rich in 9,12-C18:2 fatty acid, to 7,10-C16:2 fatty acid, 5,8-C14:2 fatty acid, 9,12-C18:2 fatty alcohol, 7,10-C16:2 fatty alcohol, and 5,8-C14:2 fatty alcohol. 7,10,13-C16:3 fatty acid, 9,12,15-C18:3 fatty alcohol, and 7,10,13-C16:3 fatty alcohol were also converted from linseed oil, which is rich in 9,12,15-C18:3 fatty acid, by the A. hydrophila isolates. These fatty acids and fatty alcohols are rarely found in natural oils. Since decreasing fatty acid carbon chain lengths from the carboxyl end and reducing unsaturated fatty acids to unsaturated fatty alcohols are both difficult reactions to accomplish by chemical means, we suggest that these A. hydrophila isolates may facilitate introduction of new bioprocess for producing rare unsaturated fatty acids and rare unsaturated fatty alcohols, especially fatty alcohols harboring more than two double bonds.     

Direct Real-Time PCR with Ethidium Monoazide: A Method for the Rapid Detection of Viable Cronobacter sakazakii in Powdered Infant Formula

The goal of this study was to establish a rapid assay for the specific detection of viable Cronobacter sakazakii in powdered infant formula (PIF). Samples were subjected to treatment multiple times with ethidium monoazide with a concentration gradient (gEMA) prior to PCR to discriminate viable from dead C. sakazakii cells. To improve the current detection limits, we developed a new buffer for direct quantitative real-time PCR (DqPCR) without DNA isolation. Using 17 PIF samples, our rapid assay was compared with the new U.S. Food and Drug Administration (FDA) method published in the Bacteriological Analytical Manual in 2012. Although both the new FDA method and our rapid assay, which consists of DqPCR combined with gEMA (gEMA-DqPCR), produced negative results for all 17 PIF samples, 5 of the 17 PIFs were positive by DqPCR when they were not treated with EMA. Furthermore, for PIF samples artificially contaminated with viable C. sakazakii, gEMA-DqPCR successfully detected between 1 and 9 CFU of viable C. sakazakii in 300 g of PIF within 9 h, including a 6-h preincubation. Our results indicate that multiple EMA treatments are required to avoid false-positive results due to the contamination of commercial PIF with dead or injured C. sakazakii cells. Our rapid assay may also improve the sensitivity of the screening portion required by the new FDA method published in the Bacteriological Analytical Manual in 2012.     

Pile Bearing Capacity Factors and Soil Crushabiity

The existence of large magnitude stresses at the tip of a bearing pile is a well known phenomenon leading to crushing of soil grains and thus affecting pile behavior. Classical foundation design calculations which assume that the soil fails in shear and neglect volume change can be safely used where stress levels or particle strengths prevent crushing, however in the case of weak grains or high foundation stresses consideration should be given to the effects of grain crushing and the resulting volumetric compression. Model pile tests have been carried out in two skeletal carbonate sands and a standard silica sand with the aim of examining the variation of skin friction and end bearing capacities with degree of penetration. The mobilization of the strength of crushable soils requires a much higher strain level while at the same time the end bearing pressure on the model piles exceeded 10?MPa inducing considerable particle breakage. The peak skin friction for all sands occurred at a settlement normalized by pile diameter, S/D, of less than 0.1. At this point the carbonate sands generally had lower skin friction values than the silica sand. Further displacement caused a rapid decrease in skin friction for all three materials. At higher lateral stresses the less crushable Toyoura silica sand generated higher skin frictions. Samples of Chiibishi sand were sectioned and photographed. It was observed that a spherical plastic zone was formed at the base of the pile which expanded with increasing S/D and a degraded layer of broken particles developed around the pile as S/D increased. Large values of the Marsal particle breakage factor were restricted to a zone extending outwards to one pile radius. An end bearing capacity modification factor has been proposed to adapt the conventional bearing capacity equation for soil crushability. This modification factor is a function of soil compressibility and degree of penetration. The factor was shown to decrease with increasing soil compressibility and increase with normalized penetration S/D.     

A probabilistic static fatigue model for transverse cracking in CFRP cross-ply laminates

This paper presents a delayed-fracture model for transverse cracking in CFRP cross-ply laminates under static fatigue loading. First, a delayed-fracture model for a crack in a brittle material was established on the basis of the slow crack growth (SCG) concept in conjunction with a probabilistic fracture model using the three-parameter Weibull distribution. Second, the above probabilistic SCG model was applied to transverse cracking in cross-ply laminates under static fatigue loading. The stress and the length of the unit element in the transverse layers were calculated with the aid of a shear-lag analysis, taking the residual stress into account. The transverse crack density was expressed as a function of applied stress and time with the parameters in the Paris law and the Weibull distribution function specified, in addition to the mechanical and geometrical properties. Unknown parameters were determined from experiment data gathered in static tensile and static fatigue tests. The reproduced transverse crack density at various applied loads agreed well with the experiment results.     

Structural characterization of recombinant human erythropoietins by fluorophore-assisted carbohydrate electrophoresis

Two enzyme immunoassay (EIA) systems were compared for their ability to detect Borrelia burgdorferi sensu lato specific IgG and IgM antibodies and to differentiate between symptomatic (83 patients with neuroborreliosis) and asymptomatic seropositive subjects (80 healthy controls). Antibody concentrations were determined by EIA; the antigens used were either a sonicate of B. burgdorferi or three recombinant borrelial proteins: the 14-kDa flagellin fragment, the outer surface protein C (22 kDa) and the high molecular mass protein p83 (83 kDa). In the sonicate, EIA, IgG or IgM antibodies to B. burgdorferi, or both, were detected in all patients with neuroborreliosis and in all controls. Pre-absorption of sera with Treponema phagedenis sonicate diminished the sensitivity of detection of borrelial specific IgG (IgG or IgM or both) antibodies in patients with neuroborreliosis from 80 to 57% (100 to 82%) and in the controls from 100 to 32% (100 to 37%). While being specific for B. burgdorferi, the recombinant EIAs proved to be significantly more sensitive than the sonicate EIA: IgG or IgM, or both antibodies against any of the recombinant antigens were detected in 92% of patients with neuroborreliosis and in 24% of controls. The increase in sensitivity in patients with neuroborreliosis was mostly due to the higher detection rate of IgM antibodies in the recombinant EIA (77% versus 48% in the sonicate EIA), while IgG antibodies were demonstrated with similar frequencies in both EIA systems (57% versus 60%). It was concluded that the recombinant EIAs are superior to the sonicate EIA with pre-absorption of cross-reactive antibodies in the confirmation of an acute borrelial infection and in the differentiation between symptomatic and asymptomatic infections.     

Theoretical studies of the mechanism of ethylene polymerization reaction catalyzed by diimine-M(II) (M = Ni,Pd and Pt) and Ti- and Zr-chelating alkoxides

We have analyzed the computational results for several elementary reactions of the ethylene polymerization process catalyzed by an alternative (to the existing metallocene catalysts) “non-cyclopentadienyl” catalysts such as diimine-M(II) (where M = Ni and Pd) and chelating bridged Ti- and Zr-complexes. The obtained data have been compared with those for the existing zirconocene-based catalysts. In general, it was shown that: (i) the resting stage of the process is a metal-olefin-alkyl complex, the olefin coordination energy of which is a few kcal/mol larger for diimine-M(II) systems than zirconocene or dialkoxide systems; (ii) the rate-determining barrier is a migratory insertion barrier calculated from the metal-olefin-alkyl complex, which is found to be a few kcal/mol larger for the diimine-M(II) system compared to the Cp₂ZrCH ₃ ⁺ catalyst. The presence of certain flexible bridging ligands X in the Ti-alkoxide complex, [Y(Ph)X(Ph)Y]TiCH ₃ ⁺ , which are capable of donating electron density to the cationic metal center at various stages during the reaction makes this barrier a few kcal/mol smaller for the dialkoxide than the Cp₂ZrCH ₃ ⁺ catalyst. It was shown that an increase in the metal-bridge interaction decreases the migratory insertion barrier and, consequently, increases the catalytic activity of these complexes. Although the diimine-M(II) catalysts are less active than zirconocene-based ones, the microstructure of the polymers produced by the former catalyst, which is found to be a function of temperature, ethylene, steric bulkiness of the auxiliary ligands, and transition metal center, makes them attractive for practice. We also have studied the mechanisms of several chain termination/transfer reactions, as well as the role of steric effects in the studied elementary reactions. We have clearly demonstrated tremendous possibilities of the computational chemistry in solving complex problems of the homogenous catalyst, and its high capability of predicting new and more active catalysts for different commercially important processes including olefin polymerization reactions. This revised version was published online in June 2006 with corrections to the Cover Date.     

Structure-Based Design,Synthesis, and Biological Evaluation of Imidazo[4,5-b]pyridin-2-one-Based p38 MAP Kinase Inhibitors: Part 1

We identified a lead series of p38 mitogen-activated protein kinase inhibitors using a structure-based design strategy from high-throughput screening of hit compound 1 . X-ray crystallography of 1 with the kinase showed an infrequent flip of the peptide bond between Met109 and Gly110, which was considered to lead to high kinase selectivity. Our structure-based design strategy was to conduct scaffold transformation of 1 with maintenance of hydrogen bond interactions with the flipped hinge backbone of the enzyme. In accordance with this strategy, we focused on scaffold transformation to identify imidazo[4,5-b]pyridin-2-one derivatives as potent inhibitors of the p38 MAP kinase. Of the compounds evaluated, 21 was found to be a potent inhibitor of the p38 MAP kinase, lipopolysaccharide-induced tumor necrosis factor-α (TNF-α) production in human monocytic leukemia cells, and TNF-α-induced production of interleukin-8 in human whole blood cells. Herein we describe the discovery of potent and orally bioavailable imidazo[4,5-b]pyridin-2-one-based p38 MAP kinase inhibitors that suppressed cytokine production in a human whole blood cell-based assay.     

Thermal decomposition of alkaline-earth metal hydride and ammonia borane composites

We demonstrate a method to improve the promising hydrogen storage capabilities of ammonia borane by making composites with alkaline-earth metal hydrides using ball-milling technique. The ball-milling for the mixtures of alkaline-earth metal hydride (MgH₂ or CaH₂) and ammonia borane (AB) yields a destabilization compared with the ingredient of the mixture, showing the hydrogen capacity of 8.7 and 5.8 mass% at easily accessible dehydrogenation peak temperatures of 78 and 72 °C, respectively, without the unwanted by-product borazine. Through detailed analyses on the dehydrogenation performance of the composite at various ratios in the hydride and AB, we proposed a different chemical activation mechanism from that in the LiH/AB and NaH/AB systems reported in a previous literature.