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.     

Transmittance and cathodoluminescence of AlN ceramics sintered with Ca3Al2O6 as sintering additive

Aluminum nitride ceramics were prepared by sintering with 0–4.8 mass% of Ca₃Al₂O₆ (C₃A) as a sintering additive. The transmittance in the range of 260–550 nm increased with increasing amount of C₃A. The cathodoluminescence intensity attributed to oxygen-induced defects decreased with increasing amount of C₃A. From the results, the increase of the transmittance in the range of 260–550 nm was considered to be related to the decrease of the oxygen-induced defect density.     

Passivation effects of deuterium exposure on boron-doped CVD homoepitaxial diamond

The deuterium (hydrogen) passivation effect on acceptors in boron-doped CVD homoepitaxial diamond was studied by electrical (Hall-effect) and secondary ion mass spectroscopy (SIMS) measurements. Deuterium was incorporated into the samples using microwave (MW) deuterium plasma at 673 K for 2–24 h. We observed the progress of acceptor passivation with p-type conduction, which finally resulted in a highly resistive state.     

Three-dimensional display of the bronchi using helical CT

The purpose of this study was to evaluate the clinical usefulness of three-dimensional (3D) images of the bronchi obtained using helical CT. Thirteen patients with lung cancer, one with tracheal diverticulum, and one with bronchial amyloidosis were examined. The CT scanner employed was the Toshiba Xforce. The helical CT scan cycle consisted of 20 continuous rotations, each requiring 1.5 sec, for a total scanning time of 30 sec. Scans were obtained using a 5-mm X-ray beam width, a 5-mm/1.5 sec couchtop sliding speed, and a 2-mm reconstruction interval. 3D images were reconstructed using a CEMAX VIPstation. The optimal lower and upper threshold CT values for 3D images of the bronchi were -650 and -100 HU, respectively, and 3D images clearly depicted endobronchial lesions. Cartilage crescents were also demonstrated, but longitudinal and circular mucosal folds could not be visualized. In conclusion, 3D images of the bronchi acquired using helical CT were useful in evaluating endobronchial lesions.     

Formation of toxic aldehydes in cod liver oil after ultraviolet irradiation

Formaldehyde, acrolein, malonaldehyde (MA), acetaldehyde and propanal produced from cod liver oil upon ultraviolet irradiation (λ_max=300 nm) were derivatized into nitrogenor sulfur-containing compounds and then analyzed by capillary gas chromatography with a nitrogen-phosphorus detector or a flame photometric detector. Acrolein and MA were formed at levels of 10.9±3.06 and 190.2±38.4 nmol/mg of fish oil, respectively. Maximum levels of formaldehyde, acetaldehyde and propanal formed were 7.0±0.90, 49.1±4.5 and 35.8±4.0 nmol/mg of oil, respectively. Formation of propanal in large quantities corresponded to the high content of ɛ-3 fatty acids in cod liver oil.     

A new type of arc plasma reactor with 12-phase alternating current discharge for synthesis of carbon nanotubes

A new type of arc plasma reactor with 12-phase alternating current (AC) discharge for synthesis of carbon nanotubes (CNTs) is proposed. A couple of six discharge electrodes by which have mutually electrical connection between them to enlarge the high-temperature regions in the reactor are arranged to three-dimensional locations. A new method of CNTs fabrication by this reactor, which accomplishes to enlarge the suitable growth region in high purity and at high yield, was developed.     

Generation of adaptive gait patterns for quadruped robot with CPG network including motor dynamic model

This paper describes the generation of adaptive gait patterns using new Central Pattern Generators (CPGs) including motor dynamic models for a quadruped robot under various environments. The CPGs act as the flexible oscillators of the joints and adjust joint angles to required values. The CPGs are interconnected with each other and sets of their coupling parameters are adjusted by a genetic algorithm so that the quadruped robot can realize stable and adequate gait patterns. Generation of gait patterns results in the formation of the CPG networks suitable for the formation of not only a straight walking pattern but also of rotating gait patterns. Experimental results demonstrate that the proposed CPG networks are effective for the automatic adjustment of the adaptive gait patterns for the tested quadruped robot under various environments. Furthermore, the target tracking control based on image processing is achieved by combining the general gait patterns. © 2006 Wiley Periodicals, Inc. Electr Eng Jpn, 155(1): 35–43, 2006; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20225