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.     

Theoretical model for flash generation

Reduction of flash generated in a gas vent is of great concern for manufacturers of electronic parts. The present study proposes a theoretical model for flash generation through consideration of flow characteristics in a gas vent. The model predicts the factors controlling flash, i.e., material parameters such as zero‐shear viscosity, crystallization temperature, thermal conductivity, and heat capacity, and process parameters such as injection and mold wall temperatures, packing pressure, and the clearance of a gas vent. On the other hand, we measure the amount of flash generated in the molding of poly(phenylene sulfide) (PPS) composites containing glass fiber and spherical fillers (CaCO₃ or Al₂O₃). Flash reduces with decreasing size of spherical fillers. These experimental data are successfully interpreted using the flash model. Polym. Eng. Sci., 45:198–206, 2005. © 2005 Society of Plastics Engineers     

Communication network protocol for real-time distributed controland its LSI implementation

A new token-passing mechanism, priority token passing, which features real-time access and fast detection and recovery of transmission errors, is discussed in detail in comparison with standard token-passing protocols, and its large-scale integration (LSI)-oriented design concept is described. Priority token passing includes only a small performance overhead, due to its switching functions, which can change network topology from ring to broadcast medium. A token-holding node passes the token to another node after determining the successor through priority comparison. Errors occurring during token passing can, thus, be detected and corrected simply and promptly. Priority token passing has a simple hardware implementation, requiring only small additions to the frame control circuitry, and has a small implementation overhead. The priority token-passing protocol and two other important network communication functions, dual ring network reconfiguration and high-level data link control (HDLC) normal response mode-based message transmission, are designed as a single finite-state machine, and implemented into a compact LSI chip. This integrated instrument network (IINET) chip provides complete network communication services and requires only three additional external electronic components for operation     

Effective lightning protection schemes for distribution line

The causes of lightning outage are subdivided into direct lightning strokes and induced lightning strokes, which are identified by the characteristics of the lightning overvoltage. In the past, lightning protection devices were directed mainly toward the latter, and attention has been focused on the installation of lightning protection devices, ground wires, and reinforcement of insulators. However, lightning outages continue to occur. Thus it is extremely important to clarify the fault characteristics of lightning surges and to study the effectiveness of various lightning protection devices by considering both direct lightning stroke and induced lightning stroke in order to prevent lightning outage in the future. In this research, the electromagnetic transients program (EMTP) has been applied to the direct lightning stroke, and the induced lightning outage analysis program for multiple conductor systems has been applied to the induced lightning stroke to study the effectiveness of lightning protection devices provided by combination of various lightning protection devices. The most effective lightning protection schemes are analyzed and evaluated based on verification tests on the full scale models as well as economic considerations.     

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.