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.     

A comparative study of DPSK and OOK WDM transmission over transoceanic distances and their performance degradations due to nonlinear phase noise

We have compared experimentally the transmission performance of return-to-zero differential phase-shift keying (RZ-DPSK) with RZ-ON-OFF keying (OOK), nonreturn-to-zero differential phase-shift keying (NRZ-DPSK), and NRZ-OOK for 100/spl times/10-Gb/s transmission with a spectral efficiency of 0.22 b/s/Hz over transoceanic distances. The Q degradation of the RZ-DPSK after transmission over 9180 km was 3 dB greater than that of RZ-OOK. The experimental results clearly showed the major cause of degradation for DPSK is not cross-phase modulation but self-phase modulation. The calculated nonlinear phase noise, i.e., the Gordon-Mollenauer effect, agreed with the experimental results. A distributed-Raman-amplifier assisted erbium-doped-fiber-amplified transmission line acted well in reducing the nonlinear phase noise.     

Solution intercalation of polystyrene and the comparison with poly(ethyl methacrylate)

Polystyrene nanocomposites have been prepared via solution intercalation method. Combination of wide-angle X-ray diffraction and thermogravimetric analysis is used to study the effect of solvents on the morphology of the nanocomposites as a function of the amount of residual solvent. d-Spacing of the polystyrene nanocomposites has a minimum value when the residual solvent concentration is between 2 and 4 wt%. Different interaction level between the solvent molecules and polymer chains is considered to be the reason for this special d-spacing change behavior. By comparing the solution intercalation of polystyrene and poly(ethyl methacrylate) from different solvents, it is concluded that the interactions between polymer-surfactant, solvent-surfactant, and polymer-solvent play important role for the solution intercalation of polymers.     

Suppression control method for torque vibration of three-phase HB-type stepping motor utilizing feedforward control

This paper proposes a method to reduce the vibration of the three-phase HB-type stepping motor with cogging torque by the feedforward compensation control. The compensation signal to suppress the vibration of the motor frame is obtained by the repetitive controller installing an online Fourier transformer and utilizing an acceleration sensor attached to the motor frame or an acoustic sensor such as a microphone placed close to the frame. The sensor is used only for the acquisition of the feedforward compensation data. The feedforward compensation signal at an arbitrary operating point is derived from the amplitude and phase data of the frequency components and the operating point data. Compensation data obtained by the repetitive controller is applied to the operating point changed by reference frequency and load condition in steady state. The compensation signal for the new operating point will be generated from compensation data utilizing polynomial equation approximation and linear interpolation method. The effectiveness of this proposed method is confirmed by the experimental results.     

Digitally tunable optical filters using arrayed-waveguide grating(AWG) multiplexers and optical switches

Two novel configurations for digitally tunable optical filters based on arrayed-waveguide grating (AWG) multiplexers are described in detail with emphasis on the connection of the AWG multiplexer and optical switches. Performance comparisons show that conventional configurations are disadvantaged by the switch size required and loss imbalance among the optical frequency-division-multiplexed (FDM) channels; the proposed configurations require only O(√(N)) switch elements to select one of N FDM channels, and the loss imbalance is lower by up to 75% in decibel     

Tuning-current splitting network for three-section DBR lasers

A simple current-splitting network for DBR-laser frequency tuning is proposed that distributes a single control current among two tuning sections and can adjust the current distribution without yielding impedance mismatching. The nonlinear characteristics of the tuning sections realise the non-proportional current splitting suitable for continuous tuning; 470 GHz (3.8 nm) tuning range is achieved     

Continuous hydrolysis of olive oil by lipase in microporous hydrophobic membrane bioreactor

Continuous hydrolysis of olive oil byCandida cylindracea’s lipase was studied in a microporous hydrophobic membrane bioreactor. Olive oil and buffer solution, fed continuously through two compartments partitioned by membrane, caused reaction at the interface of lipase-adsorbed membrane and buffer solution. Fatty acid was obtained in a single phase without being mixed with components of other phases. At all mean residence times, countercurrent flow mode was superior to cocurrent one. The lipase was adsorbed onto the membrane, and its adsorption was suggested to be partially specific from the experiments with enzymes having various levels of purity. The percent hydrolysis depended hyperbolically on the interfacial enzyme concentration. The hydrolysis seemed to be limited by diffusion of fat or fatty acid through the micropores of the membrane at higher interfacial enzyme concentrations. The lipase was stabilized significantly by glycerol added to the buffer solution. Satisfactory performance of the membrane bioreactor was obtained in a longterm continuous operation which lasted for 24 days by feeding buffer-glycerol (18.0%) solution over the adsorbed lipase. The operational half-life of the adsorbed enzyme was 15 days at 40 C.     

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.