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.     

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.     

Fabrication of lotus-type porous carbon steel by the continuous zone melting method and its mechanical properties

A lotus-type porous carbon steel with cylindrical pores was fabricated by the continuous zone melting method in a pressurized mixture of hydrogen and helium gases. The porosity increases with increasing partial pressure of the hydrogen gas, while the pore diameter remains almost constant, independent of the pressure. The ultimate tensile strength of the specimen with cylindrical pores parallel to the tensile direction is lower than the estimated value, assuming that the strength is decreased in proportion to the decrease of the cross-sectional area of the specimen, while the yield strength is higher than the estimated value. The compressive yield strength is also higher than the estimated value. The increase in yield strength is considered to be due to precipitation strengthening. The tensile strength is increased by quenching and tempering, while the elongation decreases. Such mechanical properties are discussed in terms of the microstructural analysis.     

A low-resistance self-aligned T-shaped gate for high-performancesub-0.1-μm CMOS

This paper describes the high performance of T-shaped-gate CMOS devices with effective channel lengths in the sub-0.1-μm region. These devices were fabricated by using selective W growth, which allows low-resistance gates smaller than 0.1 μm to be made without requiring fine lithography alignment. We used counter-doping to scale down the threshold voltage while still maintaining acceptable short-channel effects. This approach allowed us to make ring oscillators with a gate-delay time as short as 21 ps at 2 V with a gate length of 0.15 μm. Furthermore, we experimentally show that the high circuit speed of a sub-0.1-μm gate length CMOS device is mainly due to the PMOS device performance, especially in terms of its drivability     

Design for Consecutive Testability of System-on-a-Chip with Built-In Self Testable Cores

This paper introduces a new concept of testability called consecutive testability and proposes a design-for-testability method for making a given SoC consecutively testable based on integer linear programming problem. For a consecutively testable SoC, testing can be performed as follows. Test patterns of a core are propagated to the core inputs from test pattern sources (implemented either off-chip or on-chip) consecutively at the speed of system clock. Similarly the test responses are propagated to test response sinks (implemented either off-chip or on-chip) from the core outputs consecutively at the speed of system clock. The propagation of test patterns and responses is achieved by using interconnects and consecutive transparency properties of surrounding cores. All interconnects can be tested in a similar fashion. Therefore, it is possible to test not only logic faults but also timing faults that require consecutive application of test patterns at the speed of system clock since the consecutively testable SoC can achieve consecutive application of any test sequence at the speed of system clock.     

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).     

Crack opening displacement of a through-wall crack in a plate subjected to bending load

This study was performed in order to clarify crack opening displacement (COD) of through-wall cracks in a plate subjected to bending load. The former COD evaluation methods were mainly developed corresponding to tensile load, but there has been nothing that has been developed corresponding to bending load. Therefore, the authors evaluated CODs of the through-wall cracks in plates which were subjected to a bending load using finite element method (FEM) analyses, and proposed a simplified COD evaluation method accounting for both tensile and bending loads. The proposed method is useful for leakage evaluation at a crack opening of an elbow crown or in the vicinity of the coolant surface of a vessel in which the bending stress is relatively large.     

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.

     

Automated camerawork for capturing desktop presentations

A novel automated camera control method for capturing desktop presentations is introduced. Typical features and the camerawork of shots that appear frequently in TV programs are discussed. To realise those features in this automated video capturing system, the purpose of camerawork is classified from two points of view: target and aspect-of-target. Then, the correspondence between the classification and typical shots and camerawork is considered. A virtual-frame control algorithm based on this idea is proposed, and its implementation in a video production system. Results are shown that verify this method through two kinds of experiments, virtual video capturing using CG animations and real video capturing of real presentations.