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.     

Design and fabrication of RESURF MOSFETs on 4H-SiC(0001), (112~0), and 6H-SiC(0001)

Design and fabrication of lateral SiC reduced surface field (RESURF) MOSFETs have been investigated. The doping concentration (dose) of the RESURF and lightly doped drain regions has been optimized to reduce the electric field crowding at the drain edge or in the gate oxide by using device simulation. The optimum oxidation condition depends on the polytype: N/sub 2/O oxidation at 1300/spl deg/C seems to be suitable for 4H-SiC, and dry O/sub 2/ oxidation at 1250/spl deg/C for 6H-SiC. The average inversion-channel mobility is 22, 78, and 68 cm/sup 2//Vs for 4H-SiC(0001), (112~0), and 6H-SiC(0001) MOSFETs, respectively. RESURF MOSFETs have been fabricated on 10-/spl mu/m-thick p-type 4H-SiC(0001), (112~0), and 6H-SiC(0001) epilayers with an acceptor concentration of 1/spl times/10/sup 16/ cm/sup -3/. A 6H-SiC(0001) RESURF MOSFET with a 3-/spl mu/m channel length exhibits a high breakdown voltage of 1620 V and an on-resistance of 234 m/spl Omega//spl middot/cm/sup 2/. A 4H-SiC(112~0) RESURF MOSFET shows the characteristics of 1230 V-138 m/spl Omega//spl middot/cm/sup 2/.     

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.     

Noncontact Material Identification and Distance Measurement Using Effective Capacitance With CdS Cells

We propose a novel noncontact sensing method for material identification and for detection of the distance between the sensor and the surface of a material by using its electrical and optical properties. In the proposed method, capacitance between the terminals of a pair of CdS cells, called the effective capacitance, is measured, and several capacitance values are obtained by changing the emission strength of the light emitting diode, thus changing the resistance value of the CdS cells. From these values, it is possible to identify the materials and to detect the distance between the CdS cells and the material. In this paper, we compare four kinds of effective capacitance measurement methods using a pair of CdS cells and describe the proposed measurement method. In addition, this paper discusses the measurement accuracy of effective capacitance values in the proposed method and demonstrates its ability to identify six material samples-clear, white, and black acrylic, clear vinyl chloride, brown bakelite, and aluminum-and to detect the distance between the CdS cells and the material.     

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     

Correlates and prognosis in relation to intellectual dysfunction in a community-residing elderly population

To estimate the risk factors for intellectual dysfunction and examine its prognosis in a community-residing (non-institutionalized) elderly population, a randomly selected sample of 1,473 elderly people aged 65 years and over living in S city, Osaka Prefecture, was studied in October 1992, and data were obtained from 1,383, a response rate of 93.9%. A cohort of 1,383 was followed for 42 months and follow-up was completed for 1,300 (94.0%). The main results were as follows: 1) The prevalence of intellectual dysfunction did not differ significantly between sexes, and there was an increasing prevalence of intellectual dysfunction with age in both sexes. The prevalence of severe intellectual dysfunction was found to increase highly at age 85 and over. 2) By univariate analysis, odds ratios for age older than 75 years, low Activities of Daily Living (ADL), urinary and fecal incontinence, and no participation in social activities were significantly higher than 1 in any level of mild, moderate, and severe intellectual dysfunction. In the multivariate analysis using logistic regression, age older than 75 years and urinary and fecal incontinence showed significant higher odds ratios than 1 for severe intellectual dysfunction, and low ADL and treatment for hypertension also showed significant higher odds ratios than 1 for moderate intellectual dysfunction. 3) From analysis using the Kaplan-Meier method, the cumulative survival rates decreased with a decline in intellectual functioning in both age groups of 65-74 and 75 years and older. 4) Application of the Cox proportional hazards model resulted in adjusted hazard ratio for severe intellectual dysfunction of 1.79 (95% confidence interval, 1.02-3.12), controlling for other factors such as sex, age, general health status, incontinence and social activities.