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.     

Volumetric Change and Surface Properties of Temperature-Sensitive Polyvinylalcohol (PVA) Hydrogel

Polyvinylalcohol (PVA) polymer gel is a temperature sensitive polymeric gel, with a critical transition temperature (with H₂O) of 310 K. At higher than 310 K, this temperature sensitive polymer gel shrinks because of discharging water, whereas at lower temperatures, the gel swelled because of absorbing water. The reversibility of the gel's volume change was confirmed by temperature swing. The adsorption behavior of an organic compound onto the PVA polymer gel in water was tested at various temperatures. The amount of adsorbed organic compound increased remarkably at temperatures higher than about 310 K. Then, it was confirmed that the organic compound in PVA polymer gel could be reversibly adsorbed and desorbed by a temperature change between 293 and 323 K. The mechanism of adsorption of the organic compound onto the PVA polymer gel could be explained by hydration and dehydration of polymer gel.     

Relation between encoder and syndrome former variables and symbol reliability estimation using a syndrome trellis

We derive a linear correspondence between the variables of an encoder and those of a corresponding syndrome former. Using the derived correspondence, we show that the log-likelihood ratio of an information bit conditioned on a received sequence can be equally calculated using the syndrome trellis. It is shown that the proposed method also applies to recursive systematic convolutional codes which are typical constituent codes for turbo codes. Moreover, we show that soft-in syndrome decoding considering a priori probabilities of information bits is possible in the same way as for Viterbi decoding based on the code trellis. Hence, the proposed method can be applied to iterative decoding such as turbo decoding. We also show that the proposed method is effective for high-rate codes by making use of trellis modification.     

1-GHz-spaced 16-channel arrayed-waveguide grating for a wavelength reference standard in DWDM network systems

We fabricated a 1-GHz-spaced 16-channel arrayed-waveguide grating (AWG) by using a new AWG configuration where the path of each arrayed waveguide winds backward and forward across a 4-in diameter wafer without crossing any other waveguides. The ultra-narrow (< 1 GHz) and stable transmission bands of this AWG can be used to construct a wavelength reference standard covering the S, C, and L bands in the dense wavelength-division-multiplexing network systems whose frequency deviation is /spl plusmn/160 MHz.     

Low-loss athermal silica-based lattice-form interleave filter with silicone-filled grooves

We describe a low-loss athermal silica-based interleave filter with a lattice-form structure. We athermalize the filter by using a silicone-filled groove formed in each delay line and we employ a low-loss design to reduce the accumulated groove excess loss in the multiple delay lines. By using these designs, we obtained a very low groove excess loss of 0.4 dB and achieved satisfactory temperature insensitivity and optical characteristics with this filter.     

Extremely low-loss 1.5%-/spl Delta/ 32-channel athermal arrayed-waveguide grating multi/demultiplexer

A silica-based 1.5%-/spl Delta/ 100 GHz-spacing 32-channel athermal arrayed-waveguide grating (AWG) with compact size and extremely low insertion loss is described. By reducing the fibre coupling loss and the excess loss in a silicone-filled groove, an insertion loss of 1.3 dB was achieved with this athermal AWG.     

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.