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.     

High-performance self-aligned SiGeC HBT with selectively grown Si/sub 1-x-y/Ge/sub x/C/sub y/ base by UHV/CVD

Si/sub 1-x-y/Ge/sub x/C/sub y/ selective epitaxial growth (SEG) was performed by cold-wall, ultrahigh-vacuum chemical vapor deposition, and the effects of incorporating C on the crystallinity of Si/sub 1-x-y/Ge/sub x/C/sub y/ layers and the performance of a self-aligned SiGeC heterojunction bipolar transistor (HBT) were evaluated. A Si/sub 1-x-y/Ge/sub x/C/sub y/ layer with good crystallinity was obtained by optimizing the growth conditions. Device performance was significantly improved by incorporating C, as a result of applying Si/sub 1-x-y/Ge/sub x/C/sub y/ SEG to form the base of a self-aligned HBT. Fluctuations in device performance were suppressed by alleviating the lattice strain. Furthermore, since the B out diffusion could be suppressed by incorporating C, the cutoff frequency was able to be increased with almost the same base resistance. A maximum oscillation frequency of 174 GHz and an emitter coupled logic gate-delay time of 5.65 ps were obtained at a C content of 0.4%, which shows promise for future ultrahigh-speed communication systems.     

Development and exploitation of a multipurpose CFD tool for optimisation of microbial reaction and sludge flow.

A numerical analysis technique for optimisation of microbial reaction and sludge flow has been developed in this study. The technique is based on the 3D multiphase Navier-Stokes solver with turbulence models. In order to make numerical analyses of the total processes in wastewater treatment plants possible, four numerical models, the microbial reaction model, a sludge settling model, oxygen mass transfer model from coarse bubbles, and a model from fine bubbles, are added to the solver. All parameters included in those models are calibrated in accordance with experimental results, and good agreements between calculated results and experimental results are found. Finally, this study shows that the numerical technique can be used to optimise wastewater treatment plants with an example of the operational optimisation of an intermittent agitation in anoxic reactors by coarse bubbles. With a proper appreciation of its limit and advantages, the exploitation of the CFD efficiently leads us to the right direction even though it is not quantitatively perfect.     

Impact of crosstalk in an arrayed-waveguide multiplexer onN×N optical interconnection

The impact of crosstalk in an arrayed-waveguide N×N wavelength multiplexer is investigated precisely in relation to its application to wavelength-routing N×N all optical networks. In such systems multiple crosstalk light which has the same wavelength as the signal results in signal-crosstalk beat noise. We confirm that the noise is Gaussian and obtain the relation between crosstalk and power penalty. It is shown that the crosstalk must be less than -38 dB for a 16×16 system to keep the power penalty below 1 dB at a bit error rate of 10^-9     

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.     

A Model of the Relationship Between Psychosocial Variables and Diurnal Cortisol Rhythm Under Chronic Stress by Using Structural Equations

By using structural equations, we investigated the effect of chronic stress on salivary cortisol rhythm and proposed a causal model of chronic stress by using psychosocial and physiological data. First, 111 healthy workers (48 males, 63 females) completed questionnaires on chronic stress and lifestyle habits. Then, they provided saliva samples and answered questionnaires that were prepared to assess their psychological states 5 times (on waking up and at 10:00, 11:40, 14:00, and 16:00) on workdays. Structural equation modeling (SEM) revealed that chronic stress and longer commuting time resulted in sleep irregularities and this disrupted the cortisol circadian rhythm. This suggests that chronic stress disrupts the cortisol circadian rhythm even in healthy individuals, and sleep regularity mediates the effect of chronic stress on the cortisol rhythm.

     

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.     

Integrated-optic double-ring resonators with a wide free spectralrange of 100 GHz

Integrated-optic double-ring resonators with a wide free spectral range (FSR) of 100 GHz are fabricated using GeO₂-doped silica waveguides with a high relative refractive index difference (Δ) of 1.5%. The resonators are composed of two ring resonators comprising small ring waveguides with radii of 1.75 and 2.0 mm. The double-ring resonator module exhibited a wide FSR of 98.0 GHz, a finesse of higher than 138, a low crosstalk of less than -11.7 dB, and a low insertion loss of 6.1 dB. The measured FSR of 98.0 GHz is wider than any previously reported ring resonator composed of optical waveguides