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.     

Nickel‐Catalyzed Suzuki–Miyaura Coupling of a Tertiary Iodocyclopropane with Wide Boronic Acid Substrate Scope: Coupling Reaction Outcome Depends on Radical Species Stability

We describe a nickel‐catalyzed Suzuki–Miyaura arylation of a tertiary iodocyclopropane with arylboronic acids; this is an efficient and convergent strategy for providing various enantioenriched arylcyclopropanes with a quaternary stereogenic center. This is the first metal‐catalyzed coupling between a tertiary alkyl electrophile and a wide range of aromatics, including heteroaromatics. We found that the outcome of the Ni‐catalyzed coupling with halides as electrophiles was dependent on the stability of the radical species formed during the reaction. The use of tert‐butyl alcohol (t‐BuOH) as the reaction solvent was very effective, because of its stability under the radical‐generating reaction conditions.

     

The fracture toughness of inorganic glasses

Measuring the fracture toughness (K_Ic) of glasses still remains a difficult task, raising experimental and theoretical problems as well. The available methods to estimate K_Ic are reviewed, with emphasis on their respective advantages and drawbacks. In view of our current understanding, this analysis gives precedence to the SEPB method. The ultimate glass strength, the critical flaw size, and the indentation load for the onset of crack initiation are discussed, in the light of the fundamentals of fracture mechanics and classical background regarding the mechanics of brittle materials. Analytical expressions were further proposed to predict the fracture energy and fracture toughness of glasses from different chemical systems from their nominal compositions. The theoretical values were compared with the experimental ones, as obtained by self‐consistent methods when available. The agreement observed in most cases suggests that measured K_Ic values correspond to the crack propagation regime (as opposed to the crack initiation threshold), and supports previous investigations in glasses and ceramics, which showed that a crack tip is nearly atomically sharp in these materials (but for metallic glasses). Some ideas to design tougher glasses are finally presented.     

Design of new catalysts for ecological high-quality transportation fuels by combinatorial computational chemistry and tight-binding quantum chemical molecular dynamics approaches

Recently, we introduced a concept of combinatorial chemistry to computational chemistry and proposed a new method called “combinatorial computational chemistry”, which enables us to perform a theoretical high-throughput screening of catalysts. In the present paper, we reviewed our recent application of our combinatorial computational chemistry approach to the design of new catalysts for high-quality transportation fuels. By using our combinatorial computational chemistry techniques, we succeeded to predict new catalysts for methanol synthesis and Fischer–Tropsch synthesis. Moreover, we have succeeded in the development of chemical reaction dynamics simulator based on our original tight-binding quantum chemical molecular dynamics method. This program realizes more than 5000 times acceleration compared to the regular first-principles molecular dynamics method. Electronic- and atomic-level information on the catalytic reaction dynamics at reaction temperatures significantly contributes the catalyst design and development. Hence, we also summarized our recent applications of the above quantum chemical molecular dynamics method to the clarification of the methanol synthesis dynamics in this review.     

Fabrication and piezoelectric properties of BaTiO3/BaTiO3-Bi(Mg1/2Ti1/2)O3-BiFeO3 composites

We fabricated xBaTiO₃ (BT)/(1-x)[BaTiO₃-Bi(Mg_1/2Ti_1/2)O₃-BiFeO₃] (BT-BMT-BF)?+?0.1?wt%MnCO₃ composites by spark plasma sintering and investigated the effect of BT content x, BT powder size, and BT-BMT-BF composition on piezoelectric properties. For xBT/(1-x)(0.3BT-0.1BMT-0.6BF) +?0.1?wt%MnCO₃ (x?=?0–0.75) composites with a 0.5-µm BT powder, the dielectric constant was increased with x, and the relative density was decreased at x?=?0.67 and 0.75, creating optimum BT content of x?=?0.50 with a piezoelectric constant d₃₃ of 107?pC/N. When a larger 1.5-µm BT powder was utilized for the composite with x?=?0.50, the d₃₃ value increased to 150?pC/N due to the grain size effect of the BT grains. To compensate for a compositional change from the optimum 0.3BT-0.1BMT-0.6BF due to partial diffusion between the BT and 0.3BT-0.1BMT-0.6BF grains, a 0.5BT/0.5(0.275BT-0.1BMT-0.625BF)?+?0.1?wt%MnCO₃ composite with the 1.5-µm BT powder was fabricated. We obtained an increased d₃₃ value of 166?pC/N. These results provided a useful composite design to enhance the piezoelectric properties.     

Acceleration of the Rate of Silver Nanoparticle Formation Using Microbubbles in a Sonochemical Process

The effect of the addition of microbubbles on the formation of silver nanoparticles produced in an ultrasonic radiation-mediated process was investigated. The addition of microbubbles with an area-based median diameter of 26.0?µm and a bubble volume density of 0.18?cm³/L significantly increased the silver nanoparticle formation rate during the sonochemical process. The size distribution of the silver nanoparticles was largely unaffected by the addition of the microbubbles. The influence of changes in the microbubble volume density on the formation of the silver nanoparticles was also investigated; it was confirmed that the rate of formation of the silver nanoparticles increased moderately with increasing volume density. The gradient in absorption spectra was approximately 30 times larger for the case when microbubbles (median diameter: 28.3?µm) were added at 0.74?cm³/L, compared with the case without microbubbles. The results suggested that the microbubbles provided reaction sites similar to cavitation bubbles in the ultrasonic reaction.     

Consideration on the Relationship Between Dielectric Breakdown Voltage and Water Content in Fatty Acid Esters

Mineral oil is commonly used as an electrical insulating oil in transformers because of its relatively high electrical insulating ability and fluidity. Considering the depletion of resources and environmental problems, however, fatty acid esters synthesized from natural plant oils are attracting attention as an environmentally friendly insulating oil. In addition, fatty acid esters such as methyl octanoate, methyl dodecanoate, 2-ethylhexyl octanoate, and 2-ethylhexyl dodecanoate have high fluidity, and also show excellent moisture tolerance against dielectric breakdown compared to mineral oil. In the present study, to clarify the reason for the superior moisture tolerance of fatty acid esters, the status of dissolved water in esters is investigated with IR spectroscopic measurements and density functional theory (DFT) calculations. It is revealed that water molecules in fatty acid esters are trapped by the ester moiety of fatty acid esters. As a result, fatty acid esters have a higher moisture tolerance against dielectric breakdown than mineral oil.     

Detection of Multiple Blockages in Parallelized Microreactors

In a plant consisting of parallelized microreactors (MRs), the product quality is lowered because of a lack of flow uniformity among them when blockage occurs. It is not practical to install sensors in every MR from the viewpoint of cost when detecting the blocked MRs. In the previous study, the multiple blockage detection (MBD) method using a small number of sensors was proposed, but its performance became low when the number of sensors decreased. Here, the conventional algorithm for MBD is improved by considering the process behavior on blockage occurrence, and the effectiveness of the improved algorithm is demonstrated through a numerical case study. The effects of flow distributor types and sensor types on the MBD performance are numerically investigated.