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.     

Creating the brain and interacting with the brain: an integrated approach to understanding the brain

In the past two decades, brain science and robotics have made gigantic advances in their own fields, and their interactions have generated several interdisciplinary research fields. First, in the ‘understanding the brain by creating the brain’ approach, computational neuroscience models have been applied to many robotics problems. Second, such brain-motivated fields as cognitive robotics and developmental robotics have emerged as interdisciplinary areas among robotics, neuroscience and cognitive science with special emphasis on humanoid robots. Third, in brain–machine interface research, a brain and a robot are mutually connected within a closed loop. In this paper, we review the theoretical backgrounds of these three interdisciplinary fields and their recent progress. Then, we introduce recent efforts to reintegrate these research fields into a coherent perspective and propose a new direction that integrates brain science and robotics where the decoding of information from the brain, robot control based on the decoded information and multimodal feedback to the brain from the robot are carried out in real time and in a closed loop.     

Tunneling Conductance Dependence of the Tunneling Spectra in Bi2Sr2CaCu2O8

We have measured the tunneling spectra in Bi₂Sr₂CaCu₂O₈ with a scanning tunneling microscope(STM) at various tip-sample distances by changing the tunneling conductance in a controlled manner. When the tunneling conductance is increased from 1×10^–9 to 1×10^–5 S, the spectra do not show changes in particular. However, the gap value decreases steeply and the asymmetric back ground density of states turns inverted V-shaped one above 6×10^–4 S. The changes in the tunneling spectra at the high tunneling conductances are explained by the enhancement of the local carrier density induced by the pressure that the STM tip applied to the sample.     

An optical instrumentation using a sensor with a sensordummyagainst noises on sensor and transmission line

This short paper discusses the method of effectively canceling equal status normal mode noise not only on a sensor line but also on a transmission line of an optical instrument using a sensor with a sensordummy resistance     

High performance of silicided silicon-sidewall source and drain (S4D) structure

A silicided silicon-sidewall source and drain (S⁴D) structure is proposed for sub-0.1-μm devices. The merit of the S⁴D structure is that the series resistance of the source and drain is significantly reduced since the silicide layer is attached very close to the gate electrode and the silicon sidewall can be doped very highly. Thus, very high drain current drive can be expected, Another advantage of this structure is that the source and drain extensions are produced by the solid-phase diffusion of boron from the highly doped silicon-sidewall. Thus, shallow extensions with very high doping can be realized. A 75-nm gate length pMOSFET fabricated with this structure is shown to exhibit excellent electrical characteristics     

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.     

热压/热变形Nd2Fe14B/α-Fe纳米双相永磁体的研究

为了制备各向异性块状Nd2Fe14B／α—Fe纳米双相永磁体，研究了热压／热变形工艺参数与样品微观组织结构、磁性能之间的关系。结果表明，饱和磁化强度Js随模压温度的升高而提高；而剩磁Jr、内禀矫顽力Hcj和最大磁能积(BH)max开始都随模压温度的升高而上升，但超过一定温度后反而降低；同时提高热压压力会使磁性能增加，而热变形温度对磁性能影响很小。热变形后样品垂直于压力方向的磁性能略高于平行于压力方向，呈现出轻微的各向异性。Nd2Fe14B／α—Fe纳米双相永磁材料在热压／热变形后没有产生晶粒的择优长大，在晶体学上仍然是各向同性的。     

Substitution of an aspartic acid results in constitutive activation of c-kit receptor tyrosine kinase in a rat tumor mast cell line RBL-2H3

The c-kit protooncogene encodes a receptor tyrosine kinase that mediates signals required for differentiation, proliferation and survival of mast cells. We have already shown the constitutive activation of c-kit receptor tyrosine kinase (KIT) in a human mast cell leukemia line (HMC-1) and a murine mastocytoma cell line (P-815). We here examined whether such constitutive activation of KIT occurred in the rat tumor mast cell line RBL-2H3 as well, which is frequently used as a tool for studying functions of mast cells. In RBL-2H3 cells, KIT was constitutively phosphorylated on tyrosine and activated in the absence of autocrine production of its ligand, stem cell factor (SCF). Sequencing analysis revealed that one of c-kit genes of RBL-2H3 cells had a point mutation, resulting in amino acid substitution of Tyr for Asp in codon 817. When rat wild-type c-kit cDNA and mutant-type c-kit cDNA encoding KITTyr817 were transfected into cells of a human embryonic kidney cell line (293T), only mutant form KITTyr817 was constitutively phosphorylated on tyrosine and activated in the absence of SCF. Since mutations at the same Asp codon constitutively activated KIT in all the human HMC-1, murine P-815, and rat RBL-2H3 cell lines, and since the incorporation of antisense oligonucleotides of c-kit messenger RNA significantly suppressed the proliferation of RBL-2H3 cells, the activating mutations in the Asp codon of the c-kit gene appeared to be involved in neoplastic growth of mast cells.