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.     

Analysis of class D inverter with irregular driving patterns

This paper presents an analysis of Class D inverter when irregular driving patterns are given to the gate drive of the switch devices. The analysis has been carried out with focusing on the waveforms, harmonics, low-frequency components, output power, and equivalent dc resistance, which are numerically analyzed and discussed. Class D inverters with six different Q factors from 0.1 to 20 are analyzed about 2/sup 16/ driving patterns for each Q. Superior four models of the six inverters are built and tested in circuit experiments. The calculated waveforms are compared to the experimental results. Both of them are agreed well in time domain and frequency domain. Analytical results show a possibility of a novel control method with irregular driving patterns. In spite of discontinuous control, the output power or voltage can be strictly changed as if continuous using the selected driving patterns in some ranges.     

Hopping and optical absorption of electrons in nano-porous crystal 12CaO·7Al2O3

Recently it has been discovered that a nano-porous main group oxide 12CaO·7Al₂O₃ (C12A7) can be converted from a wide-gap insulator to a good transparent conductor. Using ab initio modelling we explain good conductivity of this material by very small barriers for hopping of localised electrons between neighbouring positive cages. We show that optical absorption of C12A7 in infrared region and at energies higher than 2.7 eV is due to inter-cage and intra-cage electron transitions, respectively. The proposed mechanisms can be useful in further search for conducting transparent media.     

Fracture toughness of short carbon fibre reinforced thermoplastic polyimide

The fracture toughness testing of short fibre reinforced thermoplastic materials were performed. Materials tested were the polyimide resin and also that reinforced with 20 wt% or 30 wt% short carbon fibre. For introducing the initial crack, the tapping method, the sliding method and the bridge indentation method were examined. Among them, the sliding method was found to be effective for every case. The fracture tests were conducted by the three-point bending test with several loading rates. Stable crack growth was observed for the neat material while unstable fracture occurred for the reinforced materials. The critical values of the stress intensity factor at crack initiation were greater for the reinforced materials than for the neat resin. The fracture toughness of the 30 wt% reinforced material was independent of loading rate while that of 20 wt% reinforced material increased with loading rate. In order to investigate the fracture mechanisms, fractographic observations were also performed.     

Preparation and antibacterial activity of novel fluoroalkyl end-capped oligomers/silica nanocomposites-encapsulated low molecular biocides

Fluoroalkyl end-capped N-(1,1-dimethyl-3-oxobutyl)acrylamide oligomer [R_F-(DOBAA) _n -R_F] reacted with tetraethoxysilane (TEOS) and silica nanoparticles in the presence of low-molecular weight biocides such as hibitane, hinokitiol, and hinokioil under alkaline conditions to afford R_F-(DOBAA) _n -R_F/silica nanocomposites-encapsulated these biocides in excellent to moderate isolated yields. Fluoroalkyl end-capped N,N-dimethylacrylamide oligomer [R_F-(DMAA) _n -R_F] and acrylic acid oligomer [R_F-(ACA) _n -R_F]/silica nanocomposites-encapsulated hibitane were obtained under similar conditions. Dynamic light scattering measurements showed that the size of these fluorinated nanocomposites-encapsulated biocides thus obtained is nanometer size-controlled. Additionally, these fluorinated nanocomposites were shown to have a good dispersibility and stability in methanol and water. Of particular interest, these fluorinated nanocomposites-encapsulated biocides were found to have a good antibacterial activity against Staphylococcus aureus, and these nanocomposites were applied to the surface modification of traditional organic polymers such as poly(methyl methacrylate).     

Electron transport in InGaO3(ZnO)m (m=integer) studied using single-crystalline thin films and transparent MISFETs

We have investigated the characteristics of transparent metal-insulator-semiconductor field-effect transistors (MISFETs) fabricated using InGaO₃(ZnO)_m (m=integer) single-crystalline thin films as n-channel layers and amorphous alumina as gate insulator films. The MISFETs exhibit good characteristics such as insensitivity to visible light illumination, off-current as low as ∼1 nA with a positive threshold voltage of ∼3 V and on/off current ratio of 10⁵. The field-effect mobility increased from ∼1 to ∼10 cm² (V s)⁻¹ as the m-value increased. Room temperature Hall mobility also increased. However, unexpectedly these values were lower than the field-effect mobility. It is explained by existence of shallow localized state in the homologous compounds.     

Intussusception caused by a carcinoma of the cecum during pregnancy: report of a case and review of the literature

Respiratory infections are the most common infection in children. They differ remarkably according to age, bacteria and viruses. Therefore a careful history of outbreak, age, former infections, involvement of surroundings, symptoms, etc are essential. The present study included 50 children, aged between 0.3 and 12 yrs, all treated ambulatorily. 21 received brodimoprim (B) and 29 erythromycin (E). Indications were: tonsillitis, bronchitis, otitis media, sinusitis and scarlet fever. Dosages were: B was given 10 mg/kg body weight (b.w.) initially followed by 5 mg/kg b.w., once-a-day. The duration of treatment varied between 4 and 14 days (mean 8.3 days). E was given 30.50 mg/kg b.w. 3 times per day; duration 4 to 14 days (mean 8.6 days). Overall results were: in group B:12 cures, 5 improvements, 3 failures; 1 not assessable. In group E: 20 cures, 8 improvements, 1 failure. Side effects: in group B: vomiting (1), skin reaction (2), discontinuation (2); in group E: skin reaction (1), diarrhea (5), diarrhea+vomiting (1); discontinuation (2). The differences in efficacy and tolerability in the two groups are not statistically significant. The improved compliance with a single versus t.i.d. dosages has to be taken into account.