Exploring the Minimal RNA Substrate of Flexizymes

Flexizymes are tRNA acylation ribozymes that have been successfully used to facilitate genetic code reprogramming. They are capable of charging acid substrates onto various tRNAs and tRNA analogues. However, their minimal RNA substrate has not been investigated. Here we have designed fluorescently labeled short RNAs corresponding to the four, three, and two bases (4bRNA, 3bRNA, 2bRNA) at the tRNA 3′-end and explored the minimal RNA substrate of flexizymes, dFx and eFx. 3bRNA was the observed minimal RNA substrate of the flexizymes, but the efficiency of acylation of this short RNA was two to three times lower than that of 4bRNA. The efficiency of acylation of 4bRNA was comparable with that of the microhelix, a 22-base RNA conventionally used as a tRNA analogue for analyzing acylation efficiency. We also compared the efficiencies of acylation of the microhelix and 4bRNA with various acid substrates. Thanks to the short length of 4bRNA, its acyl-4bRNA products exhibited larger mobility shifts in gel electrophoresis than those exhibited by acyl-microhelix products with every substrate tested. This indicated that 4bRNA was an ideal RNA substrate for analyzing the efficiency of acylation by flexizymes.     

A 2.5-GFLOPS, 6.5 million polygons per second, four-way VLIWgeometry processor with SIMD instructions and a software bypassmechanism

A four-way very long instruction word (VLIW), 312-MHz geometry processor with peripheral component interconnect/accelerated graphic port bus bridge was implemented in a 0.21-μm, 2.5-V, three-layer-metal CMOS process. We adopted (1) a software bypass mechanism, (2) single-instruction multiple-data stream instructions, (3) four sets of floating-point multiply add and accumulate execution units, (4) special condition code registers and a branch condition generator for a clipping operation, and (5) automatic clock delay tuning methodology. As a result of these features, we achieved a performance of 2.5 GFLOPS and 6.5 million polygons per second for a three-dimensional geometry processor, which is the highest published performance as a single geometry processor. The processor is applicable to computer-aided-design systems that require very high graphics performance     

High-Field Magnetization of the Distorted Spin Frustration System Ni5(TeO3)4Br2

We performed high-field magnetization measurements, M(H), on the Ni₅(TeO₃)₄Br₂ by using a pulse magnet. The Ni²⁺ ions (S = 1) of this compound form a two dimensional distorted spin frustrated system (distorted kagome lattice), and undergo a Neel transition at T _N ～29 K, due to the anisotropy. Below T _N , a complex spin arrangement which contains ten spin sub-lattices was suggested by neutron scattering and electron spin resonance studies. We observed a step-like transition at H _c ～11 T when T<T _N . By contrast, at T>T _N , the field-dependent magnetization curves behaved like a straight line. The H _c is close to those obtained by previous spin resonance studies of Ni₅(TeO₃)₄Br₂ and Ni₅(TeO₃)₄Cl₂ in which a spin-flop-like transition was proposed to explain the field-dependent resonance spectra. Interestingly, the observed change in magnetization (ΔM) at H _c corresponds to nearly one-half of the magnetic moment of Ni²⁺ ion, suggesting that the observed step-like behavior possibly corresponds to a spin-flip-like transition (i.e. from antiferromagnetic state to ferrimagnetic state). In addition, in the high-field region (H>H _c ), the M(H) curves did not show plateau behavior, but exhibited the straight lines having finite slopes without any sign of saturation up to 55 T.     

Oxidative coupling of methane applying a solid oxide fuel cell system

A fuel cell system using yttria-stabilized zirconia as a solid electrolyte was applied to the oxidative coupling of methane. The most active and selective catalyst was BaCO₃ deposited on Au-electrode (anode). Optimum conditions for the reaction were examined. The pressure of methane at the anode and that of oxygen at the cathode should be high as much as possible. The optimum temperature was 1073 K. Several advantages of the method are described.     

Effect of transition elements on the properties of MC carbides in IN-100 nickel-based superalloy

The effect of nine transition elements on the morphology, distribution and composition of MC carbides (MCs) in a nickel-base superalloy, IN-100, was investigated by differential thermal analysis (DTA), microstructural observation and X-ray microanalysis. The doping with tantalum, tungsten and molybdenum caused a significant change in the morphology and distribution of MCs as well as the profile of DTA curve of IN-100. The compositions of the MC in IN-100 were TiC and (Ti_0.80Mo_0.17V_0.03) C, and the doping with niobium, tantalum and tungsten changed significantly the composition of MC. On the other hand, the doping with chromium, vanadium, hafnium and zirconium scarcely changed the composition. In addition to TiC and (Ti, Mo, V) C, zirconium- and hafnium-rich MCs were found in the zirconium- and hafnium-doped alloys, respectively. The effect of the dopants on the composition of MCs could be explained by a relationship between the metallic radius and the free energy of formation for the MC.     

Fluorescence polarization study of a salt bridge between a single-chain Fv and its antigen ribonuclease A

The interaction between a single-chain Fv (sFv) of the monoclonal antibody 3A21 and its antigen, bovine pancreatic ribonuclease A (RNase A), was studied by site-directed mutagenesis of the hypervariable regions and fluorescence polarization analysis. The affinity constants of wild-type sFv and a mutant sFv D31A (Asp31 of heavy chain was replaced by Ala) for RNase A were found to be 2.7 x 10(7) and 4.7 x 10(6) M-1 in PBS at pH 7.2 and 37 degrees C, respectively. While the affinity constant of D31A is not affected by NaCl concentration, that of wild-type sFv is almost the same as that of D31A in the presence of more than 1 M NaCl. These results demonstrate that Asp31 of the heavy chain interacts electrostatically with a positively charged amino acid residue of RNase A.     

An investigation of water-gas transport processes in the gas-diffusion-layer of a PEM fuel cell by a multiphase multiple-relaxation-time lattice Boltzmann model

In order to study water-gas transport processes in the gas-diffusion-layer (GDL) of a proton exchange membrane (PEM) fuel cell system, a multiphase, multiple-relaxation-time lattice Boltzmann model is presented in this work. The model is based on the mean-field diffuse interface theory and can handle the multiphase flows with large density ratios and various viscosities. By using the standard bounce back boundary condition and an approximate average scheme for the non-slip and wetting boundary walls, respectively, detailed liquid-gas transportation in the GDL, in which exact boundary condition is difficult to be implemented, can be simulated. Unlike most of lattice Boltzmann methods based on the Bhatnagar–Gross–Krook collision operator, the present model shows a viscosity-independent velocity field, which is very important in simulating multiphase flows where various viscosities coexist. We validate our model by simulating a static droplet on a wetting wall and compare with theoretical predictions. Then, we simulate a water-gas flow in the GDL of a PEM fuel cell and investigate the saturation-dependent transport properties under different conditions. The results are shown to be qualitatively consistent with the previous numerical and theoretical works.     

Direct bonding of CMP-Cu films by surface activated bonding (SAB) method

The chemical mechanical polishing (CMP) process is indispensable to the fabrication of Cu wiring layers in the large-scale integration (LSI). Recently, a direct bonding method with low bonding temperature is required for the CMP-Cu surface in order to obtain a narrow bonding pitch. In this study, we realized a direct bonding between CMP-Cu films by means of the surface activated bonding (SAB) method at room temperature. The critical vacuum pressure to obtain large bonding strength was estimated at about 4×10⁻³ Pa from the growth rate of oxide on an active surface measured by the X-ray photoelectron spectroscope (XPS). The films were bonded successfully at the vacuum pressure better than around 3×10⁻³ Pa with the shear strength larger than 50 MPa. The transmission electron microscope (TEM) observation showed that the polycrystalline films with the mean surface roughness of 0.3 nm were bonded directly between Cu grains in atomic level. Moreover, the adhesion between the films was improved due to the stress relaxation at the interface during the thermal aging test conducted at 200 and 300^∘C in the vacuum condition.