Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae.

Drug-resistance markers for yeast transformation are useful because they can be applied to strains without auxotrophic mutations. However, they are susceptible to technical difficulties, namely lower transformation efficiency and the appearance of drug-resistant mutants without the marker. To avoid these problems, we have constructed a phosphoglycerate kinase (PGK) promoter-driven YAP1 expression cassette, called PGKp-YAP1. Yeast cells containing PGKp-YAP1 were resistant to cycloheximide, a protein synthesis inhibitor, and also to cerulenin, a fatty acid synthesis inhibitor, but not to other drugs tested. The transformation efficiency of PGKp-YAP1 using cerulenin selection was comparable to that using a URA3 auxotrophic marker when low concentrations of cerulenin were used. Non-transformed drug-resistant colonies did appear on the low-concentration cerulenin plates. However, these non-transformed colonies could easily be identified, based on their cycloheximide sensitivity and/or their resistance to aureobasidin A to which the transformants were sensitive. Therefore, the dual drug resistance of PGKp-YAP1 could be used as an effective selection for PGKp-YAP1 recipient cells. The PGKp-YAP1 marker was used to disrupt the LYS2 gene and to transform an industrial yeast strain, indicating that this marker can be used for efficient and reliable gene manipulations in any Saccharomyces cerevisiae strain.     

R & D Needs - Drying of Ceramics

New research and development needs for the drying of ceramics are prooosed. The researches on the ceramic drying are briefly reviewed and the role of the drying in the ceramic production process iS summarized. The drying must deal with molded materials and the problem is of significant difference from another drying with particulate materials. It means that the drying must be performed with keeping the molded feature and the insufficient completion of drying and the careless operation influence directly on the product quality The importnance of the R & D on the heat and moisture transfer. shrinking mechanism. the deformation behavior and the strain-stress formation in the molded clay issuggested. The subjects are also pointed out for the further improvement of the ceramic drying process and the precise design of molding with high quality.     

Molecular Simulation Study for CO2 Clathrate Hydrate

The present work has concentrated on the structure of CO₂ hydrate in the NPT ensemble using SPC (simple point charge) intermolecular potential model of water by the Monte Carlo (MC) molecular simulation. A mixture of water and CO₂ placed arbitrarily in a cubic cell has been used as a model system to simulate the CO₂ clathrate hydrate at temperatures ranging from 150–280 K and pressure up to 10 MPa. The result shows that the obtained MC simulation agrees well with the results obtained by molecular dynamic (MD) simulation. The present work is also directed to the study of structure with TIP4P potential model of water.     

Drying-Induced Strain and Stress: A Review

This paper is to review the works on strains and suesses in materials during drying.The strains and suesses are caused when temperature and moisture gradients are generated in mterials whose volume changes with heating and moisture removal. In such materials. failure and irregular deformion may be generated which affect considerably the qudity of the products after drying. In the first part. modeling procedure is introduced for the analysis of the strain-stress behavior in elaslic. viscous and visccelastic materials combined with heat and moisture transfer. An overview of the works on swains and stresses and drying characteristics are presented for malerials such as porous media. clay. sol-gels. agricultural products and foods in the second part. There are some materials that show both elasticity of the solid phase and viscosity of the fluid phase ( water or solvena∥ or viscoelasticiry. The suesses are often correlated with a suction pressure of fluids in pores and the flow rate is based on Dacy's equation for the elastic and viscous tnedia and a kind of viscoelastic media. The general canstitulive equalions. for suains and svesses are often analyzed with the stain behavior given by a function of moisturr for some media ai well. The emohasis is on the inuoduclion of comprehensive criteria for undersunding the problems of strain and stress development in materials subjected to drying.     

Stability of metastable tetragonal ZrO2 in compound powders and nucleation arguments

ZrO₂-SiO₂ (11) mixtures and ZrO₂ particles were prepared by a sol-gel method from the solutions of ZrOCl₂·8H₂O (ZOC) + Si(OC₂H₅)₄ (TEOS) + C₂H₅OH and ZOC + C₂H₅OH + H₂O + NH₄Cl systems, respectively. Quantitative changes of phase crystallized by heating were compared with those of ZOC + TEOS + H₂O and/or ZrO(NO₃)₂ · 2H₂O + H₂O systems, respectively. The stability of metastable tetragonal (mt)-ZrO₂ particles depends on the Young's modulus of the SiO₂ matrix. Transition metal oxides existing on the interface assisted in stabilizing mt-ZrO₂. The order of the assistance agreed with that of the relative field strengths of their oxides. A relationship between the ionic radius and the volatility of anionic groups, and the difficulty of nucleation for the martensitic transformation accompanying a shear stress, is suggested.     

Gas chromatographic separation of wax esters based on the degree of unsaturation

The wax esters of sperm whale head oil have been characterized by gas-liquid chromatography on an APOLAR 10C column according to their carbon number and number of double bonds. The novel technique permits the direct quantitative analysis of saturated and unsaturated wax esters.     

Electro-oxidation of dimethyl ether on Pt/C and PtMe/C catalysts in sulfuric acid

The electro-oxidation of dimethyl ether (DME) on PtMe/Cs (Me = Ru, Sn, Mo, Cr, Ni, Co, and W) and Pt/C electro-catalysts were investigated in an aqueous half-cell, and compared to the methanol oxidation. The addition of a second metal enhanced the tolerance of Pt to the poisonous species during the DME oxidation reaction (DOR). The PtRu/C electro-catalyst showed the best electro-catalytic activity and the highest tolerance to the poisonous species in the low over-potential range (<0.55 V, 50 °C) among the binary electro-catalysts and the Pt/C, but at the higher potential (>ca. 0.55 V, 50 °C), the Pt/C behaved better than PtRu/C. The apparent activation energy for the DOR decreased in the order: PtRu/C (57 kJ mol⁻¹) > Pt₃Sn/C (48 kJ mol⁻¹) ≈ Pt/C (46 kJ mol⁻¹). On the other hand, the activation energy for the MOR showed a different turn, decreased in the following order: Pt/C (43 kJ mol⁻¹) > Pt₃Sn/C (35 kJ mol⁻¹) ≈ PtRu/C (34 kJ mol⁻¹). The temperature dependence of the DOR was greater than that of the oxidation of methanol (MOR) on the PtRu/C.     

Intermittent Hypoxia Increased the Expression of DBH and PNMT in Neuroblastoma Cells via MicroRNA-375-Mediated Mechanism

Sleep apnea syndrome (SAS), characterized by recurrent episodes of oxygen desaturation and reoxygenation (intermittent hypoxia (IH)), is a risk factor for hypertension and insulin resistance. We report a correlation between IH and insulin resistance/diabetes. However, the reason why hypertension is induced by IH is elusive. Here, we investigated the effect of IH on the expression of catecholamine-metabolizing enzymes using an in vitro IH system. Human and mouse neuroblastoma cells (NB-1 and Neuro-2a) were exposed to IH or normoxia for 24 h. Real-time RT-PCR revealed that IH significantly increased the mRNA levels of dopamine β-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) in both NB-1 and Neuro-2a. Western blot showed that the expression of DBH and PNMT in the NB-1 cells was significantly increased by IH. Reporter assays revealed that promoter activities of DBH and PNMT were not increased by IH. The miR-375 level of IH-treated cells was significantly decreased relative to that of normoxia-treated cells. The IH-induced up-regulation of DBH and PNMT was abolished by the introduction of the miR-375 mimic, but not by the control RNA. These results indicate that IH stress increases levels of DBH and PNMT via the inhibition of miR-375-mediated mRNA degradation, potentially playing a role in the emergence of hypertension in SAS patients.     

Arraying needle-like TiO2 particles in a composite film by applying ac bias and its transmittance anisotropy

In an effort to improve energy-efficient windows, we experimented with a precursor slurry composite by using needle-like TiO₂ particles as the filler in a urethane matrix. Applying dc bias to the slurry failed to array the needle-like particles in the composite and to deposit on the film surface because of electrophoretic movement. However, applying ac bias of ±5 V to the precursor slurry composite for 12 h resulted in the needle-like TiO₂ particles being arrayed in the composite in a direction normal to the film surface. This resulted in an improvement in the energy efficiency of the material through an angular dependence of transmittance in the visible–near-infrared range.