Detection of pathogenic bacteria by using zinc finger protein fused with firefly luciferase

We constructed a novel bacterial genome detection system using zinc finger protein (ZF) fused with firefly luciferase (ZF-luciferase). Taking advantage of the direct recognition of double-stranded DNA (dsDNA) by ZF, we previously constructed bacterial genome detection systems that did not require dehybridization processes. To detect polymerase chain reaction (PCR) products rapidly and with a high sensitivity, we constructed two kinds of ZF-luciferase, Sp1-fused luciferase (Sp1-luciferase), and Zif268-fused luciferase (Zif268-luciferase). ZF-luciferase not only maintains luciferase activity but also shows dsDNA-binding ability and specificity. Furthermore, we succeeded in the detection of 10 copies of the genome of Legionella pneumophila and Escherichia coli O157. ZF-luciferase would be a useful tool for highly sensitive detection of pathogenic bacterial genome.     

Studies of self-organization processes in nanoporous alumina membranes by small-angle neutron scattering

We performed studies of the self-organization processes in nanoporous alumina membranes at initial and late stages of aluminum anodization by using scanning electron microscopy (SEM) and small-angle neutron scattering (SANS). SEM observations indicated three stages in the self-organization of nanopores in alumina: (1) nucleation of random nanopores with a broad radius distribution, (2) narrowing the radius distribution and (3) slow evolution of the nanoporous structure towards ordering of nanopores into large domains. SANS studies revealed orientational correlation between ordered domains of nanopores, which is characterized by a small misorientation angle. For the samples with high aspect ratios of nanopores, the SANS patterns showed azimuthal smearing, which was attributed to the redistribution of nanopores between the domains during their growth.     

Nanofork for single cells adhesion measurement via ESEM-nanomanipulator system

In this paper, single cells adhesion force was measured using a nanofork. The nanofork was used to pick up a single cell on a line array substrate inside an environmental scanning electron microscope (ESEM). The line array substrate was used to provide small gaps between the single cells and the substrate. Therefore, the nanofork could be inserted through these gaps in order to successfully pick up a single cell. Adhesion force was measured during the cell pick-up process from the deflection of the cantilever beam. The nanofork was fabricated using focused ion beam (FIB) etching process while the line array substrate was fabricated using nanoimprinting technology. As to investigate the effect of contact area on the strength of the adhesion force, two sizes of gap distance of line array substrate were used, i.e., 1 μm and 2 μm. Results showed that cells attached on the 1 μm gap line array substrate required more force to be released as compared to the cells attached on the 1 μm gap line array substrate.     

Fatigue limit of carbon and CrMo steels as a small fatigue crack threshold in high-pressure hydrogen gas

The fatigue limit properties of a carbon steel and a low-alloy CrMo steel were investigated via fully-reversed tension-compression tests, using smooth specimens in air and in 115-MPa hydrogen gas. With respect to the CrMo steel, specimens with sharp notches were also tested in order to investigate the threshold behavior of small cracks. The obtained SN data inferred that the fatigue limit was not negatively affected by hydrogen in either of the steels. Observation of fatigue cracks in the unbroken specimens revealed that non-propagating cracks can exist even in 115-MPa hydrogen gas, and that the crack growth threshold is not degraded by hydrogen. The experimental results provide justification for the fatigue limit design of components that are to be exposed to high-pressure hydrogen gas.     

Chemical-reaction dependence of plasma parameter in reactive silane plasma

Electron temperature in a silane glow-discharge plasma, being an important plasma parameter for determining photo-induced instability in the resulting hydrogenated amorphous silicon (a-Si:H), has been studied under various film-preparation conditions. We have used an optical-emission-intensity ratio of Si^* to SiH^* (I_si*/I_siH^*) which corresponds to the high-energy-tail slope of the electron-energy-distribution function in the plasma as a measure of electron temperature in a reactive silane glow-discharge plasma. We have found quite differently from the conventional non-reactive glow-discharge plasma such as hydrogen plasma that the electron temperature in the silane plasma is strongly modified by the substrate temperature (gas temperature) especially under high silane-gas partial-pressure condition. This anomalous behavior of the electron temperature in the silane plasma has been explained by means of gas-phase-polymerization reaction and electron-attachment process to the polymers in the plasma. The electron temperature has been remarkably reduced when a hydrogen-dilution method and a cathode-heating method are used which are considered to control polymer-formation reactions in the silane plasma together with utilization of conventional electron-temperature-controlling methods such as a very high plasma-excitation frequency and an application of magnetic field for electron-confinement. As a consequence of the reduction of electron temperature in the silane plasma, highly stabilized a-Si:H has been successfully obtained even under high growth rate conditions of 1.5 nm s^-1.     

SIMULATED EFFECTIVE THERMAL CONDUCTIVITY OF SINTERED,RANDOMLY PACKED SPHERES AND STATISTICAL STRUCTURES OF PACKINGS

Effective thermal conductivity of sintered spherical particles is estimated by a computer simulation. The simulation consists of (i)simulated random packing of equal spheres by a method of "rigid sphere free fall into a virtual box," (ii) finite element method (FEM) estimation of the thermal resistance of a "sintered" pair of spheres, and (iii) simulated heat conduction tests of a "random network," as a model of sintered particles, of thermal resistors with the estimated resistance; these tests yield the effective conductivity of sintered spherical particle aggregates. Statistical structures of the random packings of spheres are examined. The random packings constructed are standard "loose random packings." The cumulative diameter distribution of circles appearing on cross-sections of the packings is in complete agreement with the theoretical prediction for all three orthogonal directions, implying that the packing structures are isotropic. And, despite this result, the zenithal distribution of branch orientations deviates from a uniform one; more spheres are in contact with their neighbors at the zenithal angles of theta approxequal 45 degrees. The effective conductivities obtained are not isotropic but transversely isotropic, which is due to the deviation of branch orientations.     

Characteristics of Two-Phase Flows in a Rectangular Microchannel with a T-Junction Type Gas-Liquid Mixer

In this study, gas–liquid two-phase flows in a horizontal rectangular microchannel have been investigated. The rectangular microchannel has a hydraulic diameter of 0.235 mm, and a width and depth of 0.24 mm and 0.23 mm, respectively. A T-junction-type gas–liquid mixer was used to introduce gas and liquid in the channel. In order to know the effects of liquid properties, distilled water, ethanol, and HFE7200 were used as the test liquids, with nitrogen gas was used as the test gas. The flow pattern, the bubble length, the liquid slug length, and the bubble velocity in two-phase flow were measured with a high-speed video camera, and the void fraction was determined from the bubble velocity data and the superficial gas velocity data. In addition, the pressure drop was also measured with a calibrated differential pressure transducer. The bubble length data were compared with the calculation by the scaling law proposed by Garstecki et al. [7]. The bubble velocity data and/or the void fraction data were well correlated with the well-known drift flux model [12] with a new distribution parameter correlation developed in this study. The frictional pressure drop data were also well correlated with the Lockhart-Martinelli method with a correlation of the two-phase friction multiplier.     

Full‐wave optoelectrical modeling of optimized flattened light‐scattering substrate for high efficiency thin‐film silicon solar cells

The flattened light‐scattering substrate (FLiSS) is formed by a combination of two materials with a high refractive index mismatch, and it has a flat surface. A specific realization of this concept is a flattened two‐dimensional grating. When applied as a substrate for thin‐film silicon solar cells in the nip configuration, it is capable to reflect light with a high fraction of diffused component. Furthermore, the FLiSS is an ideal substrate for growing high‐quality microcrystalline silicon (µc‐Si:H), used as bottom cell absorber layer in most of multijunction solar cell architectures. FLiSS is a three‐dimensional structure; therefore, a full‐wave analysis of the electromagnetic field is necessary for its optimal implementation. Using finite element method, different shapes, materials, and geometrical parameters were investigated to obtain an optimized FLiSS. The application of the optimized FLiSS in µc‐Si:H single junction nip cell (1‐µm‐thick i‐layer) resulted in a 27.4‐mA/cm² implied photocurrent density. The absorptance of µc‐Si:H absorber exceeded the theoretical Yablonovitch limit for wavelengths larger than 750 nm. Double and triple junction nip solar cells on optimal FLiSS and with thin absorber layers were simulated. Results were in line with state‐of‐the‐art optical performance typical of solar cells with rough interfaces. After the optical optimization, a study of electrical performance was carried out by simulating current–voltage characteristics of nip solar cells on optimized FLiSS. Potential conversion efficiencies of 11.6%, 14.2%, and 16.0% for single, double, and triple junction solar cells with flat interfaces, respectively, were achieved. Copyright © 2012 John Wiley & Sons, Ltd.     

Reinvestigation of the Oxidative Folding Pathways of Hen Egg White Lysozyme: Switching of the Major Pathways by Temperature Control

It has been well established that in the oxidative folding of hen egg white lysozyme (HEL), which has four SS linkages in the native state (N), three des intermediates, i.e., des[76–94], des[64–80], and des [6–127], are populated at 20 °C and N is dominantly formed by the oxidation of des[64–80] and des[6–127]. To elucidate the temperature effects, the oxidative folding pathways of HEL were reinvestigated at 5–45 °C in the presence of 2 M urea at pH 8.0 by using a selenoxide reagent, DHS^ox. When reduced HEL was reacted with 1–4 equivalents of DHS^ox, 1S, 2S, 3S, and 4S intermediate ensembles with 1–4 SS linkages, respectively, were produced within 1 min. After the oxidation, 3S was slowly converted to the des intermediates with formation of the native structures through SS rearrangement. At 5 °C, des[76–94] was populated in the largest amount, but the oxidation to N was slower than that of des[64–80] and des[6–127]. At 35 °C, on the other hand, des[64–80] and des[6–127] were no longer stable, and only des[76–94] was populated. The results suggested that the major folding pathways of HEL can be switched from one to the other by temperature control.