Breeding of a high tyrosol‐producing sake yeast by isolation of an ethanol‐resistant mutant from a trp3 mutant

A novel breeding strategy for a high tyrosol‐producing sake yeast was developed by isolating an ethanol‐resistant mutant from a tryptophan auxotrophic mutant of a sake brewery yeast. Since tyrosol has antioxidant, cardioprotective and taste‐sharpening effects, increasing the tyrosol level of alcohol beverages could be beneficial in alcohol production. Since the transporters of aromatic amino acids are degraded by several stresses and mutants defective in the synthesis of aromatic amino acids are sensitive to ethanol, it was hypothesized that the degradation of these transporters should be inhibited in ethanol resistant mutants isolated from the auxotrophic mutants of aromatic amino acids, and that the uptake of aromatic amino acids would be increased in the mutants. Consistent with this hypothesis, sake was brewed with the ethanol‐resistant mutant of a tryptophan auxotrophic mutant and the sake was found to contain a lesser content of tyrosine and a higher content of tyrosol relative to the sake brewed with the parental strains. The taste of the sake brewed with the mutant strain could be discriminated from the sake brewed with the parental strains, probably because of the altered concentrations of tyrosol and certain amino acids and organic acids. The results suggest that combining the isolation of an ethanol‐resistant mutant and an auxotrophic mutant is an effective method to breed a brewing strain with a modified metabolism of these substances. Copyright © 2012 The Institute of Brewing & Distilling     

Accumulation of paralytic shellfish poison (PSP) and biotransformation of its components in oysters, Crassostrea gigas, fed with the toxic dinoflagellate Alexandrium tamarense

As a part of our studies on the mechanism of uptake of paralytic shellfish poison (PSP) and the kinetics of its accumulation in bivalves, oysters Crassostrea gigas were experimentally contaminated with PSP by being fed with the toxic dinoflagellate Alexandrium tamarense for 2, 4, 6, 8 and 10 days. Temporal variations in the PSP contents and their profiles in oysters during the feeding experiment were monitored by high-performance liquid chromatography (HPLC) and the toxin profile of the oysters was compared with that of A. tamarense. Toxins excreted from the infested oysters into the seawater for 2 and 10 days were recovered and analyzed by HPLC. PSP toxicity rapidly appeared in the tissues of oysters and their toxicity levels reached 0.6 (0.3), 2.2 (1.1), 1.0 (0.5), 3.4 (1.6) and 1.1 (0.5) MU/g (nmol/g) shucked meat at 2, 4, 6, 8 and 10 days, respectively. The accumulation rates of toxin, calculated from the total amount (nmol) of toxins expressed by the total cell number fed during the exposure period and the toxicity of the oysters, were 14.1, 18.7, 5.1, 14.9 and 3.2% for 2, 4, 6, 8 and 10 days. During feeding experiments, the toxin profile of oysters changed substantially, showing marked differences from the proportions found in the toxigenic dinoflagellate used as food. The toxin components in this strain existed almost exclusively as beta-epimers, which accounted for 66.3 mol% of the total. This contrasts with the case of the oysters, where the beta-epimers represented 24.8, 29.8, 25.1, 27.3 and 25.2 mol% of the total at 2, 4, 6, 8 and 10 days, respectively. The amount of gonyautoxin-1 (GTX1) accumulated in oysters increased linearly and slowly for 8 days and the maximum content of GTX1 reached 51.3 mol%. The composition of GTX group compounds recovered from the seawater in which the oysters had been reared was a little different from that within the oyster tissues.     

Precise metabolic flux analysis of coryneform bacteria by gas chromatography-mass spectrometry and verification by nuclear magnetic resonance

Precise metabolic flux analysis (MFA) by gas chromatography-mass spectrometry (GC-MS) and computer calculation was performed, and the consistency of the estimated results was verified by independently performed nuclear magnetic resonance (NMR) analysis. The precise estimation of flux by the integration method of the mass isotopomer signal, defined as the coefficient of variance (CV) of multiple determination, was investigated, and the results estimated using different data sets with the same magnitude of error were confirmed. The CV of multiple determinations was sufficiently small to discuss and compare the fluxes of a metabolic pathway. The estimated fluxes using the GC-MS data were cross-validated with the NMR data that were independently measured and not used for MFA. The developed method was successfully applied to the MFA of the growth phase of two different glutamate-producing coryneform bacteria, Corynebacterium glutamicum and C. efficiens. The difference in the growth rate between these two bacterial species was discussed while considering the results of MFA, including forward and backward (exchange) fluxes.     

Development of radiochromic dosimeters based on polymer films with fluoran and divinyl phthalide dyes

Journal of Materials Science: Materials in Electronics - We fabricated polyvinyl chloride (PVC) and poly(methyl methacrylate) (PMMA) films containing NIR Black78, Black100 and Black400 for use as...     

Studies on chitin. IX. Crosslinking of water-soluble chitin and evaluation of the products as adsorbents for cupric ion

Water-soluble chitin was successfully crosslinked to varying extents with glutaraldehyde in homogeneous aqueous solutions to improve the properties as an adsorbent for metal cations, and the effects of crosslinking were discussed. Complete insolubilization was achieved with the fivefold excess aldehyde, but, in terms of adsorptivity of Cu²⁺, the chitin crosslinked at an aldehyde/amino group ratio of 1.0 was found to exhibit remarkable capacity and was much superior to others. The desorption of Cu²⁺ from the adsorption complex was also attained effectively at pH 2.0. These results indicated that the loose crosslinking was quite simple and efficient to produce high capacity adsorbents for practical use. Thermal behavior of the crosslinked chitin was examined by TMA and TGA; a softening phenomenon was observed at 145°C.     

Base metal Co-fired (Na,K)NbO3 structures with enhanced piezoelectric performance

A NaF-Nb₂O₅ flux doped (Na,K)NbO₃ (NKN) based lead-free ceramic was successfully co-fired with nickel inner electrodes in reduced atmospheres. No chemical reactions and/or inter-diffusion were detected at the interface between the nickel (Ni) electrodes and the NKN-based piezoelectrics. Dielectric, resistivity, and electromechanical performance were measured with processing under different firing conditions and flux additions to obtain high densities. Ceramics are obtained with submicron grain structures with the NaF-Nb₂O₅ sintering aids (2 and 4 wt%) fluxes, and high densities when firing at low pO₂ (10^?10 atms) atmospheres at sintering temperatures ~1150 °C for 2 hours. High resistivities and low losses can be obtained through a second annealing condition at 850 °C and 10^?7 atms at 8 hours. High d ₃₃ values (over 350 pm/V) determined under unipolar converse electromechanical measurements were obtained in the simple prototyped co-fired structures to show feasibility towards base metal electrodes in multilayer actuators.     

Fundamental Evaluation of Power Supply and Rectifiers for Wireless Power Transfer Using Magnetic Resonant Coupling

This paper provides a fundamental analysis of a power supply and rectifiers for wireless power transfer using magnetic resonant coupling (MRC). MRC enables efficient wireless power transfer over middle‐range transfer distances. MRC for wireless power transfer should operate at a high frequency in the industry science medical band, such as 13.56 MHz, because the size of the transfer device decreases at higher transfer frequencies. Therefore, the output frequency of the power supply on the transmitting side should be 13.56 MHz. In addition, the rectifier on the receiving side is operated at a high frequency. This paper focuses on the reflected power in the power supply and rectifiers. Thus, the parametric design method is clarified for the power supply, including a low‐pass filter to match the output, the impedance of the power supply with the characteristic impedance of the transmission line. In addition, the effects on the rectifiers of silicon carbide and gallium nitride diodes are confirmed by performing an experiment and a loss analysis.     

Verification of Iron Loss Calculation Method Using a High‐Precision Iron Loss Analyzer

Because of the improved performance of power devices, the volume of the ac filter inductors used in high‐frequency PWM inverters has been reduced. However, the temperature rise in the filter inductor due to this miniaturization has become more pronounced. Therefore, we have proposed an iron loss calculation method for the ac filter inductor. However, the accuracy of the value calculated via the loss map method cannot be verified, because the iron loss arising during each switching period cannot be measured with conventional power measuring instruments. In order to resolve this problem, we developed an inductor loss analyzer (ILA), which allows precise measurement of the iron loss in the inductor during each switching period. The accuracy of the calculation of iron loss in the filter inductor by the loss map method was verified with the ILA. We found that the value calculated by the loss map method differed slightly from the value measured with the ILA. However, these differences can be reduced if we take into account the accurate flux density calculation and the effect of the duty ratio of PWM pulses on the loss. Finally, we verified that the loss map method can provide accurate iron loss calculations.     

Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (p < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture. Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.