Novel poly-gamma-glutamate-processing enzyme catalyzing gamma-glutamyl DD-amidohydrolysis

The pgdS gene product of Bacillus subtilis, PgdS, cleaves poly-gamma-glutamate (PGA) in an endo-peptidase-like fashion. However, its catalytic property remains obscure. In this study, a simple assay for the PgdS enzyme using 1-fluoro-2,4-dinitrobenzene was developed, and some characteristics of PgdS, such as optimal pH, were examined. The enzyme was strongly inhibited by a thiol-modifying reagent, suggesting that it possesses essential cysteine residue(s) in catalysis. PgdS exhibited a high affinity to PGA that consisted mainly of D-glutamate residues, but no affinity to PGA composed only of L-glutamate residues (L-PGA). The enzyme processed DL-copolymer-type PGA (DL-PGA) with an average molecular mass of 1,000 kDa to a high-molecular-mass L-glutamate-rich fragment (average 200 kDa), the L-rich PGA fragment, and low-molecular-mass fragment composed mostly of D-glutamate residues (average 5 kDa), D-fragment. To deepen our understanding of the catalytic property of the PgdS enzyme, we analyzed the structures of the N- and C-terminal regions and found that D-glutamyl residues successively lie even at both ends of the L-rich PGA fragment. Our observations indicate that PgdS is a novel endo-peptidase that specifically cleaves the gamma-amide linkage between two D-glutamate residues in PGA, i.e., gamma-glutamyl DD-amidohydrolase. The enzyme is possibly useful in the biochemical processing of B. subtilis DL-PGA.     

传统大豆发酵食品的生理功能

本文综述了我国传统大豆发酵食品的各种功能性成分，总结了其生物调节功能，并针对我国传统大豆发酵食品的研究开发提出了一些建议。     

In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka

Detection of endocrine disrupting chemicals, in particular, environmental estrogens with living organisms, has many advantages if compared to chemical analysis. The screening of novel pollutants with meaningful endpoints, the integration of uptake, bioconcentration, and excretion as well as the evaluation of endocrine disrupting effects with respect to toxicity require in vivo biotests for estrogen-like substances (ELSs). Critical disadvantages of whole organism biotests are their low sensitivity and the need for laborious and time-consuming work. To overcome these problems, we have developed a transgenic medaka strain harboring the green fluorescence protein (GFP) gene driven by choriogenin H gene regulatory elements. Choriogenin H is an egg envelope protein induced by estrogens in the liver. With yolk sac larvae of this strain, GFP induction in liver was observed 24 h after onset of aqueous exposure to 0.63 nM 17beta-estradiol (E2), 0.34 nM ethynylestradiol, or 14.8 nM estrone. Furthermore, concentrated sewage treatment effluent induced GFP expression. Comparison of E2 equivalents estimated by GFP-induction in transgenic medaka, a YES assay, and GC/MS showed detection limits in the same order of magnitude. These results indicated that the sensitivity of the transgenic medaka strain was sufficient for application as an alternative model in monitoring environmental water samples for ELSs.     

Inhibitory effects of flavonoid glycosides isolated from the peel of Japanese persimmon (Diospyros kaki Fuyu) on antigen-stimulated degranulation in rat basophilic leukaemia RBL-2H3 cells

We found that two distinct flavonoid glycosides isolated from the peel of Japanese persimmon (Diospyros kaki Fuyu), isoquercitrin (Isq) and hyperin (Hyp), are capable of inhibiting antigen-stimulated degranulation in rat basophilic leukaemia RBL-2H3 cells. In order to elucidate the underlying mechanisms, we examined effects of Isq and Hyp on cellular responses induced by antigen stimulation. Treatment with both Isq and Hyp markedly inhibited antigen-stimulated elevation of intracellular free Ca²⁺ concentration and reactive oxygen species (ROS). Isq and Hyp did not affect NADPH oxidase (NOX) activity, but they possessed DPPH radical-scavenging activity similar to that of epigallocatechin gallate, a potent anti-oxidant, Finally, Isq and Hyp showed little or no effects on Ag-stimulated Syk activation or phosphorylation of signalling molecules. These results indicate that inhibition of antigen-stimulated degranulation by Isq and Hyp is mainly due to suppression of intracellular Ca²⁺ elevation, which is caused by direct scavenging of ROS that are generated by NOX. Our findings suggest that Isq and Hyp, isolated from the peel of persimmon, would be beneficial for alleviating symptoms of type I allergy.     

Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk

A rapid and sensitive method for detection of staphylococcal enterotoxin B (SEB) in raw and dry milk samples with the use of antibody-based immunomagnetic separation (IMS) in conjunction with flow cytometry (FCM) was developed. Sheep anti-SEB immunoglobulin G (IgG) was immobilized on Dynabeads M-280. The SEB initially binds to the capturing antibody, which is bound on the magnetic beads. The rabbit anti-SEB IgG binds to the captured toxin and is further labeled with a Cy5-labeled goat anti-rabbit IgG antibody. The percentage of the beads that were fluorescent was measured by FCM. FCM was carried out for 1 min, and the data obtained were expressed as histograms for particle size (forward light scatter) and histograms for fluorescence intensity. A peak corresponding to the magnetic beads was clearly distinguished from a peak derived from contaminating particles in the sample solution. In the absence of SEB, about 10% of the beads emitted fluorescence. The percentage of fluorescent beads and the fluorescence intensity increased with increasing SEB concentrations. For this IMS-FCM assay, the lower limits of detection for SEB were estimated to be 0.01 and 0.25 ng/ml for buffer and milk samples, respectively.     

The effect of the framework structure on the chemical properties of the vanadium oxide species incorporated within zeolites

V-silicalite catalysts (VS-1 and VS-2) prepared by hydrothermal synthesis have been studied by ESR, XAFS (XANES and EXAFS) and photoluminescence spectroscopy. The in situ characterization of these V-silicalites shows that vanadium is present within the zeolitic framework as a highly dispersed tetrahedrally coordinated V-oxides, VO₄ unit, having a short V=O bond length. Photoluminescence spectroscopy in static and dynamic mode, as well as XAFS studies allow to detect in the V-silicalites different V species than that present in V-HMS or V/SiO₂, in terms of V=O bond length, vibrational energy, bond angle and lifetime of the excited triplet state. It is suggested that the combined contribution of the neighboring Si---OH group attached to the VO₄ unit and the zeolitic rigid framework structure of V-silicalites cause a more significant and pronounced effect on the chemical properties of the VO₄ unit than the flexible structure of V-HMS or V/SiO₂. Moreover, the dynamic quenching of the phosphorescence by the addition of reactant molecules such as NO or propane indicates that the V species in the excited triplet state can be expected to be the active sites for the photocatalytic reactions.     

采用数值模拟方法研究同时氧化烟气中NO和SO2的可行性添加剂

Mechanism analysis on simultaneous oxidation of NO and SO2 with additives was presented and numerical simulation was developed to investigate the performances of three additives on oxidation of NO and SO2. The simulation result showed that reaction temperature, residence time, additive dose and NO concentration influence the oxidation process significantly. There exists an optimum reaction condition for each additive. n-C4H10 has the strongest ability to oxidize NO and SO2.     

Consideration of Energy Dissipation for the Strength of Adhesive Joints

For some adhesive joints where the main difference is the degree of contact at the interface, failure occurs not at the interface, but some distance away in the polymer itself. This cohesive mode of failure in the polymer was always found to be the case in our studies of cupric oxide to branched polyethylene interfaces, even where the joint was so weak that the peeled surface seemed clean of the polymer to the naked eye. It was observed that the strength of the joint was associated with the coarseness of the texture of the peeled surface of the polymer. With a differential scanning calorimetry technique we have shown that the coarseness of the surface texture and therefore the strength of the joint, is a direct function of the amount of polymer involved in plastic deformation. The strength criteria for the adhesive joint of this kind is thus the energy of deformation and not the maximum tensile stress that the material can withstand.     

Preparation of micrometer‐sized,multifunctional capsule particles for cosmetic by microsuspension polymerization utilizing the self‐assembling of phase separated polymer method

Micrometer‐sized polymer particles with encapsulated ultraviolet (UV) absorbent, fluorescent agent, and blue pigment were successfully prepared by microsuspension polymerization utilizing the Self‐assembling of Phase Separated Polymer method. The particles were characterized by optical microscope and particle size distribution analysis, and they were evaluated on their usefulness for cosmetic using UV spectrometry, colorimetry, VISIA? Evolution, and bioassay. The capsule particles had multifunctional properties, which are very attractive in the cosmetic field, especially in whitening, brightening, the improvement of face‐texture, and less noticeable pores in face, as well as the protection from UV. Moreover, bioactivities of the particles under the UV irradiation, which were examined with the films prepared from capsule components, revealed not only makeup effect but also the activation of human epidermal keratinocytes. The results suggest the importance of blue light in the field of cosmetics. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

A simple approach to observe non-conductive hydrated materials with FE-SEM: Case study on porous hydroxyapatite green bodies

The purpose of this study is to develop a simple method to observe the surface morphology of non-conductive material in its hydrated condition. Here porous hydroxyapatite (HAp) green body prepared by gelcasting process was considered as a case study, and, the resultant body was subsequently treated with hydrophilic ionic liquid (IL). The surface morphology and the pore structure of the IL-treated porous HAp green body were successfully observed even in hydrated condition without any charging using field emission scanning electron microscope (FE-SEM). Results showed that the pore diameter from 300 to 600 μm in as-prepared green body was reduced to 100–300 μm in the sample sintered at 1000 °C. Raman results showed that the IL forms weak hydrogen bond with water molecules within the sample and, that prevents from drying in vacuum condition. In addition, the IL acts as a conducting media for HAp ceramics to be observed under FE-SEM.