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.     

Fracture toughness evaluation of adjacent flow weld line in polystyrene by the SENB method

Fracture toughness of adjacent flow weld lines, defined as weld lines that occur when two flow fronts meet and continue to flow together in the same direction (meld line or hot weld line), was evaluated by the single‐edge notched‐bend (SENB) method using three differently‐shaped obstructive pins. Although the fracture toughness varied depending upon the shapes of the pin, the values could be standardized as the distance from the meeting point of the two flow fronts flowing around the pin. The fracture toughness decreased drastically from the meeting point along the weld line and then slightly increased. These characteristic features could be explained by flow‐induced molecular orientation at the weld line interface. The molecules around the meeting point that were initially oriented parallel to the weld line due to fountain flow were able to relax, and then entanglement across the weld line interface developed because the flow stopped in the middle of the filling process, resulting in high fracture toughness. In contrast, the material at the downstream side of the weld line continued flowing during the filling process, being stretched along the flow direction. So, the molecular orientation at this area could not relax. In addition, the V‐notch shape, i.e., the depth and length at the surface of the weld line, which also varied depending on the shape of the obstacles, was considered to be identical when the meeting point was allowed to be a datum point. Thus, the meeting point was found to be a significant factor when the properties of weld lines are investigated. POLYM. ENG. SCI., 45:1059–1066, 2005. © 2005 Society of Plastics Engineers     

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     

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.     

Sputter deposition technology as a materials engineering

Sputter deposition is currently being widely used in the microelectronics industry for the production of silicon integrated circuits. Recently interest has been focused on sputter deposition as a new materials processing technique. The highly energetic sputtered atoms enhance crystal growth and/or sintering during film growth. This results in lowering of the growth temperature of high temperature materials including cubic diamonds. Single crystals of complex ceramics materials could be prepared by sputter deposition through epitaxial growth process. Atomically controlled deposition using multi-target sputter enables to make man-made superlattice including high-T _C superconductors of layered perovskite. At present sputter deposition is one of key materials technologies for the coming century.     

Temperature dependence of kinetics and diffusion coefficients for ferrocene/ferricenium in ammonium-imide ionic liquids

The electrochemical behavior of ferrocene (Fc) and ferricenium (Fc⁺) was investigated by cyclic voltammetry with convolutional and semi-differential electroanalyses in the temperature range 298-373 K in the hydrophobic room temperature ionic liquid systems consisting of bis(trifluoromethanesulfone)imide anion with quaternary ammonium cation. The experimental results indicated that the redox reaction of Fc/Fc⁺ was reversible in this ionic liquid and the charge-transfer rate constant (k⁰) has been obtained. Mass transport towards the electrode is a simple diffusion process and the diffusion coefficient (D) for Fc/Fc⁺ has been also calculated. These results indicated that the k⁰ and D increased with increasing temperature in ionic liquids. The validity of the Arrhenius law was verified by investigating the temperature dependences of k⁰ and D.     

Physicochemical Properties of Palm Oil-Based Puff Pastry Model Margarines Related to Their Baking Performance in Long-Term Storage

Developing trans-free alternative fat solutions suitable for specific applications remains a challenge in edible fats and other domains. This is particularly true for palm oil-based puff pastry margarines, which suffer from post crystallization problems, leading to dramatic loss of functionality. This research is aimed at investigating the influence of triacylglycerol (TAG) compositions of palm oil-based puff pastry margarines on the physical properties of the fat crystal network, which determine the functionality of such products. Three model puff pastry margarines are produced at pilot scale under the same crystallization conditions. They share the same fatty acid composition and close solid fat content (SFC) profiles, whereas the proportions of major TAG (tripalmitoylglycerol (PPP), 1,3-di-palmitoyl-2-oleoylglycerol (POP), 1,2-di-palmitoyl-3-oleoylglycerol (PPO), 1,2-dioleoyl-3-palmitoylglycerol (POO)) are different. Polymorphism, melting profile, hardness, microscopic structures, and baking performance (puffing effect) of the model fats are examined during a period of 6 months. The following results are obtained: 1) The TAG composition significantly influences the post crystallization processes occurring in palm oil-based margarines. 2) High amounts of POP show negative influences. 3) The proportions of POP, PPO, and PPP should be carefully balanced to prevent detrimental crystal network rearrangements, leading to textural modifications (hardness increase) and significant reduction in baking performance. Practical Applications : The results presented in this work could be helpful for edible fat products developers, especially for roll-in fat applications. This research provides an overview of the relevant properties to study for the assessment of puff pastry margarine functionality. It also highlights the importance of ensuring long-term stability of palm oil-based fat products. Finally, it emphasizes that certain combinations of fat materials should be avoided to maintain the quality of palm oil-based puff pastry margarines.     

Humidity Sensor Characteristics of Woodceramics

The humidity sensor characteristics of Woodceramics were investigated. The Woodceramics used in this experiment were prepared at 650–900°C. The size of specimens selected was 1 × 1 × 10 mm . One was used as-cut and the other was polished. Above 700°C, the resistance of the specimen was below 50 and the decrease of resistance with increasing relative humidity was small. The specimen prepared at 650°C had a resistance of about 1 k. The resistance of as-cut specimens decreased with increase in relative humidity, but that of the polished specimen did not change significantly. The difference between as-cut and polished samples was explained by the differences in surface micro-structure.     

Novel process for cover glass with ideal stress distribution

Abstract — This paper proposes a new process to manufacture cover glass that overcomes a strength trade‐off between the face and the edge. In the process, alkali barrier films are deposited on glass faces before an ion exchange process in order to control face stress properties without inhibiting the edge strengthening. As a demonstration of the process, alkali‐alumino‐silicate glass sheets with sputter‐deposited SiO₂ films were chemically strengthened, and then their stress properties and strengths were investigated. As a result, thicker SiO₂ films cause lower face DOL (depth of strengthened layer), and it is observed that the faces have lower DOL than the edges. In strength tests corresponding to major fracture modes of smartphone cover glass, specimens with 80–100 nm films have more balanced face performance and better edge impact strengths than the no‐film specimen.     

Reduction of higher harmonic sequences in terminal voltage of an AC exciting synchronous generator using magnetically anisotropic slot wedges

This paper describes relationships between characteristics of a magnetically anisotropic slot wedge and voltage higher harmonics in an ac exciting synchronous generator terminal due to permeance ripple caused by an open-slot-type rotor. From the relationships obtained, practical guidelines were formulated for machine designs applying the magnetically anisotropic slot wedge. The results are as follows.

• 1 (1) Equivalent permeability of the magnetically anisotropic slot wedge became isotropic wedge permeability for air-gap flux ripple.
• 2 (2) Two ways were identified to reduce voltage-higher harmonics by using the slot wedges.
• 3 (3) The level of leakage flux passing through the anisotropic slot wedge was determined only by its permeability in the width direction.
• 4 (4) Effectiveness of the magnetic anisotropic slot wedges inserted into the stator slots was verified from experimental results using a model machine of the open- slot-type rotor.