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.     

Isolation of antimicrobial agent from the marine algae Cystoseira hakodatensis

Cystoseira hakodatensis is an unutilised brown algae belonging to family Sargassaceae. A crude methanol extract from the algae showed inhibitory effects on the growths of Bacillus cereus and Bacillus licheniformis. To isolate the major antimicrobial agent, a sequential active‐guided isolation procedure was applied: liquid–liquid extraction, column chromatography and bio‐autography. A marked antimicrobial agent (active α) was isolated in hydrophobic fraction and was determined to phenolics without carbohydrates and proteins by phytochemical test. Regarding the antimicrobial potential, the isolated active α showed better inhibitory effects against B. cereus and B. licheniformis at 2 and 4 times of lower concentrations (62.5 and 31.3 μg mL^?1) in comparison with epigallocatechin gallate. These results showed that C. hakodatensis is a potential source of antimicrobial agent capable of preventing the growth of the two bacteria.     

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.     

Liquid Marble Patchwork on Super-Repellent Surface

Liquid marble (LM) is a droplet that is wrapped by hydrophobic solid particles, which behave as a non-wetting soft solid. Based on these properties, LM can be applied in fluidics and soft device applications. A wide variety of functional particles have been synthesized to form functional LMs. However, the formation of multifunctional LMs by integrating several types of functional particles is challenging. Here, a general strategy for the flexible patterning of functional particles on droplet surfaces in a patchwork-like design is reported. It is shown that LMs can switch their macroscopic behavior between a stable and active state on super-repellent surfaces in situ by jamming/unjamming the surface particles. Active LMs hydrostatically coalesce to form a self-sorted particle pattern on the droplet surface. With the support of LM handling robotics, on-demand cyclic activation–manipulation–coalescence–stabilization protocols by LMs with different sizes and particle types result in the reliable design of multi-faced LMs. Based on this concept, a single bi-functional LM is designed from two mono-functional LMs as an advanced droplet carrier.     

Identification of stress-induced leakage current components and thecorresponding trap models in SiO2 films [MOStransistors]

Time-decay stress-induced leakage current (SILC) has been systematically investigated for the cases of both Fowler-Nordheim (FN) stress and substrate hot-hole stress. From the three viewpoints of the reproducibility of the current component for the gate voltage scan, the change of oxide charge during the gate voltage scan, and the resistance of the current component to thermal annealing, it has been found that time-decay stress-induced leakage current is composed of five current components, regardless of stress type. Trap models corresponding to each current component have been proposed. In addition, it has also been proven that holes generate the electron traps related to one of those current components     

Binary Adsorption Isotherms of Water and Ethyl Acetate Vapor for Materials Contained in a Box of a Tobacco Product

An experimental study has been carried out on the characteristics of binary adsorption isotherms of water and volatile flavor for typical materials used in a box of a tobacco product or cigarettes. Ethyl acetate chosen as a model for water-soluble volatile flavor. Binary adsorption isotherms for the tobacco, papers, filters, and activated carbons were measured with a flow-type multi-component adsorption system under the canstant conditions of temperature a t 303 K and vapor pressure of water a t 2.5 kPain the vapor pressure range of ethyl acetate from 0 to 4.2 kPa. A linear equation was applied to express the binary adsorption isotherms for the tobacco, papers and filters, while a Dubinin-Astakhov equation was applied for the activated carbons. The binary adsorption was characterized into three groups, depending on the selectivity as well as the mechanism of adsorption; i.e.(l) for tobacco and papers, water was adsorbed much greater rather than ethyl acetate, (2) for filters, ethyl acetate was adsorbed on the surface as great as water, (3) for activated carbons, ethyl acetate was adsorbed much greater onto their micropores rather than water. The results showed that ethyl     

High-dose intravenous steroid pulse therapy in thyroid-associated ophthalmopathy

MIcrospheres containing diclofenac sodium (DS) were prepared using carboxymethylcellulose (CMC) as the main support material (1.0, 2.0, 3.0% (w/v)) and aluminum chloride as the crosslinker. Drug to polymer ratios of 1:1, 1:2 and 1:4 were used to obtain a range of microspheres. The microspheres were then coated with an enteric coating material, Eudragit S-100, efficiency, % yield value, particle sizes an in-vitro dissolution behaviour were investigated. The surface of the enteric coated microspheres seemed to be all covered with Eudragit S-100 from scanning electron microscopy observation. It was also observed that increasing the CMC concentration led to an increase in the encapsulation efficiency, % yield value and particle size and decreased the release rate. Eudragit S-100 coating did not significantly alter the size but the release rate was significantly lower even when the lower concentration solution was used.     

Evolutionary design of dynamic neural networks for evaporator control

This paper presents a novel neural network (NN) to control an ammonia refrigerant evaporator. Inspired by the latest findings on the biological neuron, a dynamic synaptic unit (DSU) is proposed to enhance the information processing capacity of artificial neurons. Treating the dynamic synaptic activity after the nonlinear somatic activity helps to capture the dynamics demarcated by the Gaussian activation pertaining to the input space. This practice leads to a remarkable reduction in curse of dimensionality. The proposed NN architecture has been compared with two other conventional architectures; one with dynamic neural units (DNUs) and the other with nonlinear static functions as perceptrons. The objective is to control evaporator heat flow rate and secondary fluid outlet temperature while keeping the degree of refrigerant superheat in the range 4–7 K at the evaporator outlet by manipulating refrigerant and evaporator secondary fluid flow rates. The drawbacks of conventional approaches to this problem are discussed, and how the novel method can overcome them are presented. An evolutionary approach is adopted to optimize the parameters of the NN controllers. Then evaporator process model is accomplished as a combination of governing equations and a sub NN resulting in a simple and sufficiently accurate model. The effectiveness of the proposed dynamic NN controller for the evaporator system model is validated using experimental data from the ammonia refrigeration plant.     

Development of 170 GHz/500 kW gyrotron

A development of 170GHz/500kW level gyrotron was carried out as R&D work of ITER. The oscillation mode is TE_31,8. In a short pulse experiment, the maximum power of 750kW was achieved at 85kV/40A. The efficiency was 22%. In the depressed collector operation, 500kW/36%/50ms was obtained. The maximum efficiency of 40% was obtained at P_RF=470kW whereas the power decrease by the electron trapping was observed. Pulse extension was done up to 10s at P_RF=170kW with the depressed collector operation. The power was limited by the temperature increase of the output window.     

Construction of a control system for disturbance rejection using a dual observer

A well‐known control system which can reduce the adverse effects of disturbances is a disturbance observer. However, in many cases of mechanical systems, system disturbances which do not satisfy the matching condition may be imposed. Therefore, it may be difficult to reduce the adverse effects of the disturbances by the traditional disturbance observer. In this paper, a method of control system synthesis for disturbance rejection using a dual observer is proposed. This method is based on the zeroing induced by the disturbance localization problem. This problem may be solved by dividing the state space into observable subspace and unobservable subspace. As compared with an H_∞ controller based on perfect observation, the usefulness of the proposed control system for disturbance rejection is demonstrated by numerical simulations for a two‐mass spring system. © 2002 Scripta Technica, Electr Eng Jpn, 138(4): 50–60, 2002; DOI 10.1002/eej.1138