Identification of diluent degradation products in radiolyzed PUREX solvent

Diluent degradation products in gamma radiolyzed Plutonium URanium Extraction (PUREX) solvent were characterized, under the conditions similar to those prevailing in the hot cell for reprocessing. Alkali wash of the degraded solvent followed by vacuum distillation resulted in primary separation. Column separation of the distillation residue followed by spectroscopic analysis using GC-MS and NMR revealed the residue predominantly to be a mixture of dodecan-3-ol, dodecan-4-ol, and dodecan-5-ol in the ratio of 2:1:6, respectively and a mixture of 2-nitrododecane, 3-nitrododecane, 4-nitrododecane, and 5-nitrododecane in the ratio of 2:2:1:1, respectively. The identified compounds were confirmed by synthesizing them individually and comparing their spectral data.     

Sphingomyelin Deacylase,the Enzyme Involved in the Pathogenesis of Atopic Dermatitis,Is Identical to the β-Subunit of Acid Ceramidase

A ceramide deficiency in the stratum corneum (SC) is an essential etiologic factor for the dry and barrier-disrupted skin of patients with atopic dermatitis (AD). Previously, we reported that sphingomyelin (SM) deacylase, which hydrolyzes SM and glucosylceramide at the acyl site to yield their lysoforms sphingosylphosphorylcholine (SPC) and glucosylsphingosine, respectively, instead of ceramide and/or acylceramide, is over-expressed in AD skin and results in a ceramide deficiency. Although the enzymatic properties of SM deacylase have been clarified, the enzyme itself remains unidentified. In this study, we purified and characterized SM deacylase from rat skin. The activities of SM deacylase and acid ceramidase (aCDase) were measured using SM and ceramide as substrates by tandem mass spectrometry by monitoring the production of SPC and sphingosine, respectively. Levels of SM deacylase activity from various rat organs were higher in the order of skin > lung > heart. By successive chromatography using Phenyl-5PW, Rotofor, SP-Sepharose, Superdex 200 and Shodex RP18-415, SM deacylase was purified to homogeneity with a single band of an apparent molecular mass of 43 kDa with an enrichment of > 14,000-fold. Analysis by MALDI-TOF MS/MS using a protein spot with SM deacylase activity separated by 2D-SDS-PAGE allowed its amino acid sequence to be determined and identified as the β-subunit of aCDase, which consists of α- and β-subunits linked by amino bonds and a single S-S bond. Western blotting of samples treated with 2-mercaptoethanol revealed that, whereas recombinant human aCDase was recognized by antibodies to the α-subunit at ~56 kDa and ~13 kDa and the β-subunit at ~43 kDa, the purified SM deacylase was detectable only by the antibody to the β-subunit at ~43 kDa. Breaking the S-S bond of recombinant human aCDase with dithiothreitol elicited the activity of SM deacylase with ~40 kDa upon gel chromatography. These results provide new insights into the essential role of SM deacylase expressed as an aCDase-degrading β-subunit that evokes the ceramide deficiency in AD skin.     

Gradient-Index polymer materials and their optical devices

A gradient-index (GRIN) plastic optical fiber was prepared by a heat-drawing process of the GRIN preform rod obtained by a new interfacial-gel copolymerization technique. The attenuation loss of the GRIN fiber was 179 kB/km at 650-nm wave-length and its numerical aperture was 0.20-0.25. To attain low loss and highly dense integrated circuits, steric radial-GRIN optical waveguide components such as GRIN branching, star coupler, multi-coupler, and a 2-D GRIN lens array, were monolithically fabricated by this technique. A novel spherical GRIN sphere lens, whose index function is symmetrical about the center of the sphere, with a low spherical aberration, was prepared by a modified suspension polymerization technique. A W-shaped GRIN sphere has been newly proposed to further reduce the spherical aberration, compared with the parabolic GRIN sphere previously proposed.     

Modeling and numerical analyses of microwave plasmas for optimizations of a reactor design and its operating conditions

A model of MW plasmas has been developed, which prescribes distributions of electron temperature as well as electron number density under self-consistent electromagnetic field. In the proposed model, an energy balance equation is algebraically solved, and only electron continuity equation is essentially coupled to the Maxwell equations. Preliminary simulations are carried out and the results are compared with corresponding results obtained by using other model.     

Application of real-time polymerase chain reaction (PCR) coupled with ethidium monoazide treatment for selective quantification of viable bacteria in aquatic environment

Ethidium monoazide (EMA) was used to quantify DNA selectively from viable cells with healthy membrane/cell wall system, but not from dead cells, of a target bacterium in the aquatic environment using real-time PCR. Spiking experiments to determine the EMA treatment conditions showed that EMA treatment with EMA at 10-25 microg/ml and subsequent halogen light exposure for 2 min was suitable for selective quantification of DNA from viable cells in an aquatic sample using real-time PCR coupled with EMA treatment (real-time EMA-PCR). Optimized real-time EMA-PCR was applied in combination with culture-based method and conventional real-time PCR without EMA treatment to elucidate the behavior of an Escherichia coli strain inoculated into a pond water microcosm. Quantification results obtained using real-time EMA-PCR were lower than those by conventional real-time PCR without EMA treatment and higher than those by culture-based method. The results suggest that quantification by real-time EMA-PCR seemed to represent the viable population, which would partly include viable but non-culturable state bacteria. Real-time EMA-PCR optimized here can be a useful tool for selective monitoring of the viable population of a target bacterium in the aquatic environment, and thereby contribute to assessment of potential microbial risks generated from waterborne pathogenic bacteria.     

Small fatigue crack growth characteristics and fracture surface morphology of low carbon steel in hydrogen gas

Owing to energy conservation and environmental concerns, hydrogen has been suggested as a next-generation energy source. However, hydrogen known to seep into a metal, degrade its strength, and accelerate fatigue crack growth rates. We have investigated the effects of hydrogen gas on the small fatigue crack growth characteristics of low carbon steel JIS S10C by conducting bending fatigue tests on a specimen with a small blind hole and placed in a low-pressure hydrogen environment. The fatigue crack growth rate in hydrogen was higher than that in nitrogen. The fracture surface of the specimen in hydrogen showed intergranular facets in the low- growth-rate range and a quasi-cleavage fracture surface with brittle striations in the high-growth-rate range. The specimen only showed a ductile fracture surface for nitrogen. The small-fatigue-crack growth rate for nitrogen is given by ${dl/dN\propto \Delta \varepsilon_{p}^{n}l}$ , where l, N, and ${\Delta \varepsilon_{p}}$ represent the crack length, number of repetitions, and plastic strain range, respectively. This equation was also satisfied for hydrogen, but only over a short strain range from ${\Delta \varepsilon_t = 0.25}$ to 0.37?% in which the fracture surface exhibited intergranular facets and a ductile morphology, but no quasi-cleavage fracture. The exponent n of the equation was 1.22 in nitrogen and 0.66 in hydrogen environment. The small-fatigue-crack growth law can be used for safe material designs in hydrogen environments.     

Water‐Soluble Chiral Ruthenium(II) Phenyloxazoline Complex: Reusable and Highly Enantioselective Catalyst for Intramolecular Cyclopropanation Reactions

The intramolecular cyclopropanation of various trans‐allylic diazoacetates and alkenyl diazoketones has been achieved with excellent enantioselectivities of up to 98% ee and in quantitative yields by using a water‐soluble ruthenium(II)/hydroxymethyl(phenyl)oxazoline catalyst in a water/ether biphasic medium. The catalyst could be reused at least five times.     

Paths of progress in liquid metal processing

Industry has identified three major issues as being fundamental to future technological developments: process step elimination, product-process integration, and intelligent processing. This article reviews these concepts by discussing recent research at the University of Toronto on plasma processing, netshape casting, and diagnostic sensors for the evaluation of liquid metal quality.     

Transamination as a side-reaction catalyzed by alanine racemase of Bacillus stearothermophilus

The pyridoxal form of alanine racemase of Bacillus stearothermophilus was converted to the pyridoxamine form by incubation with its natural substrate, D- or L-alanine, under acidic conditions: the enzyme loses its racemase activity concomitantly. The pyridoxamine form of the enzyme returned to the pyridoxal form by incubation with pyruvate at alkaline pH. Thus, alanine racemase catalyzes transamination as a side function. In fact, the apo-form of the enzyme abstracted tritium from [4'-3H]pyridoxamine in the presence of pyruvate. A mutant enzyme containing alanine substituted for Lys39, whose epsilon-amino group forms a Schiff base with the C4' aldehyde of pyridoxal 5'-phosphate in the wild-type enzyme, was inactive as a catalyst for racemization as well as transamination. However, when methylamine was added to the mutant enzyme, it became active in both reactions. These results suggest that the epsilon-amino group of Lys39 participates in both racemization and transamination when catalyzed by the wild-type enzyme.     

Role of lysine 39 of alanine racemase from Bacillus stearothermophilus that binds pyridoxal 5'-phosphate. Chemical rescue studies of Lys39 --> Ala mutant

The lysine residue binding with the cofactor pyridoxal 5'-phosphate (PLP) plays an important role in catalysis, such as in the transaldimination and abstraction of alpha-hydrogen from a substrate amino acid in PLP-dependent enzymes. We studied the role of Lys39 of alanine racemase (EC 5.1.1.1) from Bacillus stearothermophilus, the PLP-binding residue of the enzyme, by replacing it site-specifically with alanine and characterizing the resultant K39A mutant enzyme. The mutant enzyme turned out to be inherently inactive, but gained an activity as high as about 0.1% of that of the wild-type enzyme upon addition of 0.2 M methylamine. The amine-assisted activity of the mutant enzyme depended on the pKa values and molecular volumes of the alkylamines used. A strong kinetic isotope effect was observed when alpha-deuterated D-alanine was used as a substrate in the methylamine-assisted reaction, but little effect was observed using its antipode. In marked contrast, only L-enantiomer of alanine showed a solvent isotope effect in deuterium oxide in the methylamine-assisted reaction. These results suggest that methylamine serves as a base not only to abstract the alpha-hydrogen from D-alanine but also to transfer a proton from water to the alpha-position of the deprotonated (achiral) intermediate to form D-alanine. Therefore, the exogenous amine can be regarded as a functional group fully representing Lys39 of the wild-type enzyme. Lys39 of the wild-type enzyme probably acts as the base catalyst specific to the D-enantiomer of alanine. Another residue specific to the L-enantiomer in the wild-type enzyme is kept intact in the K39A mutant.