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.     

Identification of stress-induced leakage current components and thecorresponding trap models in SiO2 films [MOStransistors]

Time-decay stress-induced leakage current (SILC) has been systematically investigated for the cases of both Fowler-Nordheim (FN) stress and substrate hot-hole stress. From the three viewpoints of the reproducibility of the current component for the gate voltage scan, the change of oxide charge during the gate voltage scan, and the resistance of the current component to thermal annealing, it has been found that time-decay stress-induced leakage current is composed of five current components, regardless of stress type. Trap models corresponding to each current component have been proposed. In addition, it has also been proven that holes generate the electron traps related to one of those current components     

Effect of surfactants on phenol removal by the method of polymerization and precipitation catalysed by Coprinus cinereus peroxidase

The removal of phenol by peroxidase‐catalysed polymerization was examined using Coprinus cinereus peroxidase in the presence of surfactants. The non‐ionic surfactants with poly(oxyethene) residues, Triton X‐100, Triton X‐405 and Tween 20, enhanced the phenol removal efficiency at a level similar to high relative molecular mass poly(ethylene glycol) (relative molecular mass 3000). Although the improvement in the removal efficiency was less than that of Triton X‐100, Span 20, sodium lauryl sulfate (SDS) and lauryl trimethylammonium bromide (DTAB) also enhanced the removal efficiency. The requirement of the enzyme for almost 100% removal of 100 mg dm^?3 phenol decreased to one‐fourth by the addition of 30 mg dm^?3 Triton X‐100. Triton X‐100, Triton X‐405, Tween 20 and DTAB could reactivate the enzyme precipitated with the phenol polymer, leading to the restarting of the phenol removal reaction. Copyright © 2003 Society of Chemical Industry     

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     

Preparation and properties of transparent cellulose hydrogels

Highly transparent cellulose hydrogels with physical crosslinkage were prepared from nonaqueous organic cellulose solutions and viscose by coagulating and regenerating cellulose in an aqueous solution containing a water‐miscible organic solvent. Nonaqueous organic cellulose solutions used were LiCl/dimethylacetamide, paraformaldehyde/dimethyl sulfoxide, and triethylammonium chloride/dimethyl sulfoxide. Preparation conditions and physical properties of the transparent cellulose hydrogels were studied. The transparency of the cellulose hydrogels depended on the composition of the aqueous solution containing the organic solvent. Furthermore, transparent cellulose hydrogels from viscose showed high tensile strength. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 3020–3025, 2003