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.     

Mechanical properties of SiNxCx ceramic films prepared by plasma CVD

The mechanical properties of Si₃N₄-SiC, SiN_x and SiC_y films prepared at a low temperature of 400 °C by plasma chemical vapour deposition are reported. Microhardness, internal stress of the film and adhesive strength between the film and glass or stainless steel substrate were evaluated as principal mechanical properties. Microhardness was measured to be about 10 to 20 G Pa dependent on the film composition in each system. Internal stress of the films on borosilicate glass substrates extensively varied from tensile to compressive with the film composition change from Si₃N₄ to SiC. Adhesive strength, as ascertained by the scratch test, was about 580 to 800 MPa for crown glass substrates, and about 210 to 310 M Pa for 316 stainless steel substrates. It is pointed out that tensile stress in these films brought about more abrupt decreases of the adhesive strength than did compressive stress.     

[110]-surface strained-SOI CMOS devices

We have newly developed [110]-surface strained-silicon-on-insulator (SOI) n- and p-MOSFETs on [110]-surface relaxed-SiGe-on-insulator substrates with the Ge content of 25%, fabricated by applying the Ge condensation technique to SiGe layers grown on [110]-surface SOI wafers. We have demonstrated that the electron and the hole mobility enhancement of [110]-surface strained-SOI devices amounts to 23% and 50%, respectively, against the mobilities of [110]-surface unstrained MOSFETs. As a result, the electron and the hole mobility ratios of [110]-surface strained-SOI MOSFETs to the universal mobility of (100)-surface bulk-MOSFETs increase up to 81% and 203%, respectively. Therefore, the current drive imbalance between n- and p-MOS can be reduced. Moreover, both the electron and the hole mobilities of the [110]-surface strained-SOIs strongly depend on the drain current flow direction, which is qualitatively explained by the anisotropic effective mass characteristics of the carriers on a [110]-surface Si. As a result, the [110]-surface strained-SOI technology with optimization of the current flow directions of n- and p-MOS is promising for realizing higher speed scaled CMOS.     

Hepatitis GB virus C genome in the serum of aplastic anaemia patients receiving frequent blood transfusions

GB virus C (GBV-C) RNA was detected in five of 18 patients with aplastic anaemia who had received blood transfusions, whereas it was not detected in eight patients who had not received any transfusions. Antibody against hepatitis C virus (anti-HCV) was detected in nine patients in the transfusion group, compared with one of eight who had not received any transfusions. Therefore, the route of transmission of both GBV-C and HCV in these patients appeared to have been multiple blood transfusion. Since all of the GBV-C RNA-positive patients harboured anti-HCV, GBV-C seems to frequently superinfect with HCV. Neither GBV-C nor HCV is likely to have been a causative agent of the anaemia in the cases examined.     

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.