Large-scale preparation and glycan characterization of sialylglycopeptide from bovine milk glycomacropeptide and its bifidogenic properties

Bovine glycomacropeptide (GMP) is a 7,000-Da glycopolypeptide released from κ-casein during cheese making. The O-glycan chains linked to GMP have many biological activities, but their utilization for nutraceutical products is limited due to their low content. To concentrate the functional glycan chains of GMP, we prepared sialylglycopeptide concentrate (SGC) from GMP-containing whey protein concentrate via proteolytic digestion of peptide chains and concentration of sialylglycopeptide by ultrafiltration using membranes with a molecular weight cut-off of 1,000 Da. The abundant saccharides detected in the prepared SGC were N-acetylneuraminic acid (Neu5Ac: 32.3% wt/wt), N-acetylgalactosamine (11.3%), and galactose (10.2%), which constitute O-glycans attached to GMP. The Neu5Ac content in SGC was found concentrated at approximately 4.8-fold of its content in GMP-containing whey protein concentrate (6.8%). Structural analysis of O-glycopeptides by liquid chromatography tandem mass spectrometry identified 88 O-glycopeptides. Moreover, O-acetylated or O-diacetylated Neu5Ac was detected in addition to the previously characterized O-glycans of GMP. Quantitative analysis of O-glycan in SGC by fluorescence labeling of chemically released O-glycan revealed that a disialylated tetrasaccharide was the most abundant glycan (76.6% of the total O-glycan). We further examined bifidogenic properties of SGC in vitro, which revealed that SGC served as a more potent carbon source than GMP and contributes to the growth-promoting effects on certain species of bifidobacteria. Overall, our study findings indicate that SGC contains abundant O-glycans and has a bifidogenic activity. Moreover, the protocol for the preparation of SGC described herein is relatively simple, providing a high yield of glycan, and can be used for large-scale preparation.     

Control of strain relaxation behavior of Ge1−xSnx buffer layers

We have investigated how to realize a strain-relaxed Ge_1−xSn_x layer with large in-plane lattice constant as a buffer layer for a tensile-strained Ge layer. This paper reports the dependence of strain relaxation behavior in Ge_1−xSn_x layers on the misfit strain at the interface between Ge_1−xSn_x layers and substrates. We examined control of the misfit strain by growth of Ge_1−xSn_x layers on bulk-Si and virtual Ge substrates. Large misfit strain between the Ge_1−xSn_x layer and the Si substrate leads to strain relaxation during growth and high degree of strain relaxation after annealing. However, it also leads to interfacial mixing and surface roughening with annealing. As a result, the Ge_1−xSn_x layer having a Sn content of 9.2% was achieved, and it has a potential to induce a tensile strain of 0.99% in Ge layer.     

A finite element model for simulating temperature distributions in rotating food during microwave heating

With considering the continuous rotation of the turntable during microwave heating, a three-dimensional computer model based on FEM was successfully developed to predict time-dependent temperatures distributions of food sample. The slide interface between rotating and stationary part were specially assigned and treated, as the disconnections of the nodes occur during rotating which may cause the electromagnetic analysis result in error. On this basis, temperature was estimated by coupling electromagnetic and heat transfer analysis, whereas the node coordinate and dielectric properties of sample were updated with time steps, and the heat generations were renewed according to these parameters.     

CFD analysis of the convection flow in the pan during induction heating and gas range heating

One difference between induction heating (IH) and gas range heating (GRH) is that, because the heating location of the pan is different, the convection in the pan is different. In particular, in cooking situations in which solids are mixed with liquids, such as the boiling of soup containing solid foods, the natural convection flow in the pan may affect food quality. The convection in the pan during IH and GRH was analyzed by using computational fluid dynamics (CFD), and the temperature distribution and the flow velocity distribution in the pan were obtained. These results were then compared with the measurement results. In GRH, the heat flux at the wall of the pan was obtained by experiment, and it was used for the analysis as a boundary condition. In IH, the heat generation distribution obtained by electromagnetic field analysis was used for the analysis as the boundary condition. Differences in convection flow in the pan during IH and GRH were observed by CFD modeling especially at the initial stage of heating. The simulated convection in the pan showed the same tendency as the captured movement of the tracer particle using flow visualization.     

Designed construction of recombinant DNA at the ura3Δ0 locus in the yeast Saccharomyces cerevisiae

Recombinant DNAs are traditionally constructed using Escherichia coli plasmids. In the yeast Saccharomyces cerevisiae, chromosomal gene targeting is a common technique, implying that the yeast homologous recombination system could be applied for recombinant DNA construction. In an attempt to use a S. cerevisiae chromosome for recombinant DNA construction, we selected the single ura3Δ0 locus as a gene targeting site. By selecting this single locus, repeated recombination using the surrounding URA3 sequences can be performed. The recombination system described here has several advantages over the conventional plasmid system, as it provides a method to confirm the selection of correct recombinants because transformation of the same locus replaces the pre‐existing selection marker, resulting in the loss of the marker in successful recombinations. In addition, the constructed strains can serve as both PCR templates and hosts for preparing subsequent recombinant strains. Using this method, several yeast strains that contained selection markers, promoters, terminators and target genes at the ura3Δ0 locus were successfully generated. The system described here can potentially be applied for the construction of any recombinant DNA without the requirement for manipulations in E. coli. Interestingly, we unexpectedly found that several G/C‐rich sequences used for fusion PCR lowered gene expression when located adjacent to the start codon. Copyright © 2013 John Wiley & Sons, Ltd.     

Quantum Critical Magnetization Behaviors of the Kagome- and Triangular-Lattice Antiferromagnets

The magnetization process of the S=1/2 Kagome-lattice quantum antiferromagnet is investigated using the numerical exact diagonalization up to 39-site clusters, comparing with the triangular-lattice antiferromagnet. Our previous finite-size scaling analysis with the rhombic clusters indicated the “magnetization ramp” as a novel field-induced quantum phase transition at 1/3 of the saturation magnetization. As another possible exotic behavior, we focus on the feature of the magnetization curve at the 2/3 of the saturation. The same critical exponent analysis indicates that a different singular behavior occurs at the 2/3 magnetization of the Kagome-lattice antiferromagnet, while no anomaly in the triangular-lattice one.     

Lactobacillus helveticus SBT2171, a cheese starter,regulates proliferation and cytokine production of immune cells

Consumption of a Lactobacillus helveticus SBT2171 (LH2171)-containing cheese has been reported to exhibit immunoregulatory actions, including an increase in regulatory T cell population and reduction in proinflammatory cytokine production in mice. We examined the in vitro effects of LH2171 cells per se on immune cell function, specifically proliferation and cytokine production, which are primary reactions of the immune response. Immune cell fractions were prepared by mechanical disruption of mesenteric lymph nodes (MLN), Peyer’s patches (PP), and spleens (SP) of mice. The cell fractions were dispensed into a culture plate and stimulated with anti-CD3/CD28 antibody beads in place of antigen-presenting cells or lipopolysaccharide (LPS) in the presence or absence of heat-treated LH2171 cells and other bacterial strains for comparison. After incubation, proliferation, cytokine production, and cell viability of the immune cells were determined. The LH2171 significantly inhibited the proliferation of MLN immune cells stimulated with anti-CD3/CD28 compared with other bacterial strains. The antiproliferative potency of LH2171 was effective not only on MLN but also on PP and SP stimulated with anti-CD3/CD28 or LPS. The LH2171 also decreased LPS-stimulated IL-6 production from MLN, PP, and SP, and IL-1β production from SP, but LH2171 did not affect the viability of immune cells. The LH2171 inhibited immune cell proliferation and proinflammatory cytokine (IL-6 and IL-1β) production. The inhibitory actions were not due to cytotoxicity to immune cells, suggesting that LH2171 is a dairy Lactobacillus strain with beneficial immunoregulatory properties.     

Reconfiguration management of remanufactured products for responding to varied user needs

In the face of worsening environmental problems, the manufacturing industry is required to reduce resource consumption and environmental loads during product life cycles while responding to diversified user needs without cost increases. In this paper, we propose a reconfiguration method to remanufacture a variety of products using modules extracted from returned products of multiple generations for responding to varied user needs. With this method, we can maximize the sales amount of remanufactured products, which meet the performance requirements of each user group. We demonstrate the effectiveness of the method by applying it to photocopiers.