Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations

Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B₄ (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.     

Quantitative risk assessment using a Japanese hydrogen refueling station model

Although hydrogen refueling stations (HRSs) are becoming widespread across Japan and are essential for the operation of fuel cell vehicles, they present potential hazards. A large number of accidents such as explosions or fires have been reported, rendering it necessary to conduct a number of qualitative and quantitative risk assessments for HRSs. Current safety codes and technical standards related to Japanese HRSs have been established based on the results of a qualitative risk assessment and quantitative effectiveness validation of safety measures over ten years ago. In the last decade, there has been much development in the technologies of the components or facilities used in domestic HRSs and much operational experience as well as knowledge to use hydrogen in HRSs safely have been gained through years of commercial operation. The purpose of the present study is to conduct a quantitative risk assessment (QRA) of the latest HRS model representing Japanese HRSs with the most current information and to identify the most significant scenarios that pose the greatest risks to the physical surroundings in the HRS model. The results of the QRA show that the risk contours of 10^?3 and 10^?4 per year were confined within the HRS boundaries, whereas the risk contours of 10^?5 and 10^?6 per year are still present outside the HRS. Comparing the breakdown of the individual risks (IRs) at the risk ranking points, we conclude that the risk of jet fire demonstrates the highest contribution to the risks at all of the risk ranking points and outside the station. To reduce these risks and confine the risk contour of 10^?6 per year within the HRS boundaries, it is necessary to consider risk mitigation measures for jet fires.     

Hopping and optical absorption of electrons in nano-porous crystal 12CaO·7Al2O3

Recently it has been discovered that a nano-porous main group oxide 12CaO·7Al₂O₃ (C12A7) can be converted from a wide-gap insulator to a good transparent conductor. Using ab initio modelling we explain good conductivity of this material by very small barriers for hopping of localised electrons between neighbouring positive cages. We show that optical absorption of C12A7 in infrared region and at energies higher than 2.7 eV is due to inter-cage and intra-cage electron transitions, respectively. The proposed mechanisms can be useful in further search for conducting transparent media.     

Electron transport in InGaO3(ZnO)m (m=integer) studied using single-crystalline thin films and transparent MISFETs

We have investigated the characteristics of transparent metal-insulator-semiconductor field-effect transistors (MISFETs) fabricated using InGaO₃(ZnO)_m (m=integer) single-crystalline thin films as n-channel layers and amorphous alumina as gate insulator films. The MISFETs exhibit good characteristics such as insensitivity to visible light illumination, off-current as low as ∼1 nA with a positive threshold voltage of ∼3 V and on/off current ratio of 10⁵. The field-effect mobility increased from ∼1 to ∼10 cm² (V s)⁻¹ as the m-value increased. Room temperature Hall mobility also increased. However, unexpectedly these values were lower than the field-effect mobility. It is explained by existence of shallow localized state in the homologous compounds.     

An expert system for XPLE cable deterioration diagnosis and life forecasting

The recent increase in the demand for power in urban areas has caused changes in the way power is supplied. Underground distribution systems have been introduced for convenience in urban areas and to prevent injuries in case of a crisis. As a result, many new types of power cables are being used. One type, XPLE cables (also called CV cables) have been rapidly taking the place of conventional cables such as OF or SL cables since the 1960s because of their relatively easy installation and maintainability. However, distribution cables used under severe environmental conditions deteriorate rapidly (due mainly to water treeing), and some cables experience insulation breakdown. Therefore, many diagnostic methods have been developed, though they have not been very effective. By combining methods, a high reliability can be achieved in comparison to the use of a single method. However, processing the data is difficult for on-site personnel, so many experts are required. But there is a lack of qualified experts, hence we have turned to the development of expert systems.     

Radiation tolerance for the SMASH-series front-end chips for a silicon micro-strip detector

The radiation tolerance for prototype front-end chips designed for a silicon micro-strip detector was examined with a ⁶⁰Co irradiation source to establish an allowable range of the radiation dose. The irradiated front-end chips were SMA²SH-64A, a 64-channel preamplifier array; SMA²SH-1, a single-channel preamplifier circuit with a comparator; and Control-C, a digital-control chip for the preamplifiers.     

Partial scan design and test sequence generation based on reduced scan shift method

This paper presents a partial scan algorithm, calledPARES (PartialscanAlgorithm based onREduced Scan shift), for designing partial scan circuits. PARES is based on the reduced scan shift that has been previously proposed for generating short test sequences for full scan circuits. In the reduced scan shift method, one determines proch FFs must be controlled and observed for each test vector. According to the results of similar analysis, PARES selects these FFs that must be controlled or observed for a large number of test vectors, as scanned FFs. Short test sequences are generated by reducing scan shift operations using a static test compaction method. To minimize the loss of fault coverage, the order of test vectors is so determined that the unscanned FFs are in the state required by the next test vector. If there are any faults undetected yet by a test sequence derived from the test vectors, then PARES uses a sequential circuit test generator to detect the faults. Experimental results for ISCAS'89 benchmark circuits are given to demonstrate the effectiveness of PARES.     

Complexity of the color appearance of objects with nonuniform illumination

An inversion relationship is clarified between lightness and brightness, and also between chroma and colorfulness under nonuniform illumination with different illuminance levels (high and low) within the same visual field. Brightness (or colorfulness) of object color with low lightness (or chroma) under high illuminance level is perceived higher than that of another object color with high lightness (or chroma) under low illuminance level. Two color images are given for showing the inversion phenomena on brightness–lightness and colorfulness–chroma between object colors under different illuminance levels. These color images are useful for making researchers on color understand the differences in concept between brightness and lightness, and also between colorfulness and chroma. In particular, the concept of colorfulness is important, but difficult to understand. © 2007 Wiley Periodicals, Inc. Col Res Appl, 32, 372–377, 2007     

Dissolution of Ti wires in sulphuric acid and hydrochloric acid solutions

Ti wire electrodes were immersed in acidic solutions containing H₂SO₄ and HCl of various concentrations at 353 K to evaluate corrosion rate by measurement of electric resistance change (resistometry). Addition of hydrochloric acid to sulphuric acid solution promoted depassivation of Ti. After depassivation, the immersion potential dropped to the hydrogen evolution potential and a hydride layer was formed on the surface. The hydride layer dissolved continuously in the acidic solution. SEM observation showed that Ti wires dissolved almost uniformly in the early stage and that the dissolution then trace became irregular due to nonuniform growth of the hydride layer. Dissolution rate of a Ti wire was estimated almost accurately by resistometry.