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.     

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.     

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.     

Further Characterization of 394-GHz Gyrotron FU CW GII with Additional PID Control System for 600-MHz DNP-SSNMR Spectroscopy

A 394-GHz gyrotron, FU CW GII, has been designed at the University of Fukui, Japan, for dynamic nuclear polarization (DNP)-enhanced solid-state nuclear magnetic resonance (SSNMR) experiments at 600-MHz ¹H resonant frequency. After installation at the Institute for Protein Research (IPR), Osaka University, Japan, a PID feedback control system was equipped to regulate the electron gun heater current for stabilization of the electron beam current, which ultimately achieved stabilization of output power when operating in continuous wave (CW) mode. During exploration to further optimize operating conditions, a continuous tuning bandwidth of approximately 1 GHz was observed by varying the operating voltage at a fixed magnetic field. In the frequency range required for positive DNP enhancement, the output power was improved by increasing the magnetic field and the operating voltage from their initial operational settings. In addition, fine tuning of output frequency by varying the cavity cooling water temperature was demonstrated. These operating conditions and ancillary enhancements are expected to contribute to further enhancement of SSNMR signal.     

The Direct Spectroscopy of Positronium Hyperfine Structure Using a Sub-THz Gyrotron

Positronium is an ideal system for research on Quantum Electrodynamics (QED), especially in a bound state. A discrepancy of 3.9 standard deviations has been found between the measured hyperfine structure (Ps-HFS) and the QED predictions. This may be due to the contribution of unknown new physics or common systematic effects in previous measurements, in all of which the Zeeman effect was used. We propose a new method to directly measure the Ps-HFS using a high power gyrotron. We compare two resonators which have been developed to supply sufficient power to drive the direct transition, a Fabry-Pérot resonator and a ring resonator with a diffraction grating. We plan to perform first direct measurement of Ps-HFS within the next six months.     

Efficient Excitation of Hybrid Modes in a THz Clinotron

Journal of Infrared, Millimeter, and Terahertz Waves - Various regimes of hybrid, bulk-surface, modes being excited in the clinotron oscillator with a sheet electron beam and nonuniform grating...     

Transmission Characteristics of Hybrid Modes in Corrugated Waveguides Above the Bragg Frequency

We studied the transmission characteristics of hybrid modes in a corrugated circular waveguide above the Bragg frequency to develop a broad-band transmission line for millimeter waves. Millimeter waves at 294 GHz were transmitted into a straight waveguide. From observed power profiles in waveguide cross-sections, a high attenuation rate of 0.13 dB/m was obtained. To match a theoretical attenuation constant with the experimental one, we introduced an ad hoc coefficient of conventional surface reactance in the waveguide wall. This was necessary because the wall began to look like the surface with a decreasing anisotropic reactance owing to the frequency above the Bragg frequency. Using nonlinear optimization for mode content analysis, the observed power profiles in the waveguide cross-section were matched with theoretical profiles. There was good agreement between the calculated and observed centers of power profiles and attenuation rate along the waveguide. The theoretical analysis showed that the magnetic field at the waveguide wall increases and the substantial attenuation takes place. Above the Bragg frequency coupling to backwards propagating modes is a point of consideration. A combination of the backwards propagating EH_1,26 and the forward propagating HE₁₁ modes satisfied the Bragg condition at 294.7 GHz which was the nearest frequency of operating frequency. A strong attenuation of the incoming HE₁₁ mode by Bragg resonance was not expected due to large difference of 0.7 GHz. It becomes clear that the observed high transmission loss outside of the Bragg resonance can be explained by a decrease in anisotropic surface reactance at the wall.     

Novel gold-capped nanopillars imprinted on a polymer film for highly sensitive plasmonic biosensing

Herein, a nanoporous alumina was fabricated to use as a mold in transforming nanopillar structures onto a thin film polymer by thermal nanoimprint lithography (NIL). The size of the pores was successfully controlled by varying the applied voltages and etching time. These nanoporous structures were transferred to the Cyclo-olefin polymer (COP) film surface from the porous mold by a thermal nanoimprinting process. A plasmonic substrate was fabricated by sputtering a thin layer of gold onto this nanopillar polymer structure, and the refractive index response in a variety of media was evaluated. Finally, the biosensing capacity of this novel plasmonic substrate was verified by analysis of Human immunoglobulin and achieved a minimum detection limit of 1.0 ng/mL. With the advantages of mass production with consistent reproducibility stemming from the nanoimprint fabrication process, our gold-capped polymeric pillars are ready for the transition from academic interest into commercialization systems for practical use in diagnostic applications.