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.     

Systematic packaging design tools integrating functional and environmental consequences on product life cycle: Case studies on laundry detergent and milk

This study presents systematic packaging design tools integrating functional and environmental consequences on product life cycle. To design packaging for sustainability, the trade-offs between functional and environmental aspects of packaging throughout the product life cycle should be considered. However, it is difficult for packaging designers to understand the overall trade-offs because the extent of the design consequences on the entire life cycle of packaging and its contents is unclear. We developed two tools for packaging design: the Life Cycle Association Matrix (LCAM) and the Function Network Diagram (FND). The following three steps, based on literature reviews and interviews with industrial experts, were applied. Firstly, we listed the product functions and design variables related to the functions as the attributes allocated to the product life cycle. Secondly, the attributes were connected appropriately based on causal relationships. Lastly, we identified the factors to support decision making in the packaging design procedure. As a result, the LCAM depicts the design consequences on the life cycle, and the FND determines the stakeholders affected by the design consequences. Two case studies were demonstrated to analyze the trade-offs by using our tools. In the case studies, a liquid laundry detergent bottle and a milk carton were redesigned. The tools identified the design consequences and stakeholders affected by the redesign of the usability and protective function for the detergent and milk cases, respectively. The results showed the significance of understanding the design consequences on the product life cycle by integrating the functional and environmental aspects.     

Principled analytic classifier for positive-unlabeled learning via weighted integral probability metric

Machine Learning - We consider the problem of learning a binary classifier from only positive and unlabeled observations (called PU learning). Recent studies in PU learning have shown superior...     

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.     

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.     

A cathode readout gas sampling calorimeter with conductive plastic tubes

We constructed and tested a prototype gas sampling electromagnetic calorimeter of the Pb-proportional tube sandwich type. The calorimeter uses conductive plastic tubes and cathode pad readout with a tower structure which resulted in reasonable energy and spatial resolutions for electrons in the momentum range 0.5–4.0 GeV/c; $\text{σ}{}_{\mathrm{E}}\text{E =}\text{21\%}\text{(E(}\text{GeV}\text{)}\text{)}{}^{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{, σ}{}_{\mathrm{x}}\text{= 6}\text{mm}\text{(at 3 GeV}$/c). This paper describes the test and the performance studied under various conditions.     

[110]-surface strained-SOI CMOS devices

We have newly developed [110]-surface strained-silicon-on-insulator (SOI) n- and p-MOSFETs on [110]-surface relaxed-SiGe-on-insulator substrates with the Ge content of 25%, fabricated by applying the Ge condensation technique to SiGe layers grown on [110]-surface SOI wafers. We have demonstrated that the electron and the hole mobility enhancement of [110]-surface strained-SOI devices amounts to 23% and 50%, respectively, against the mobilities of [110]-surface unstrained MOSFETs. As a result, the electron and the hole mobility ratios of [110]-surface strained-SOI MOSFETs to the universal mobility of (100)-surface bulk-MOSFETs increase up to 81% and 203%, respectively. Therefore, the current drive imbalance between n- and p-MOS can be reduced. Moreover, both the electron and the hole mobilities of the [110]-surface strained-SOIs strongly depend on the drain current flow direction, which is qualitatively explained by the anisotropic effective mass characteristics of the carriers on a [110]-surface Si. As a result, the [110]-surface strained-SOI technology with optimization of the current flow directions of n- and p-MOS is promising for realizing higher speed scaled CMOS.     

Improved direct bonding method of Nd:YVO4 and YVO4 laser crystals

We succeeded in the fabrication of bonded laser crystals composed of a neodymium-doped YVO₄ laser crystal (Nd:YVO₄) and its host crystals YVO₄ by a newly developed dry etching technique using an argon ion beam. The optical distortion caused by the bonded interface of size 5 mm × 6 mm was estimated to be 0.05λ at 633 nm. From the comparison of laser performance pumped by a laser diode, the bonded crystals could increase the laser output power by nearly twice that of the non-bonded crystals with the same degree of polarization of 99.2%. To analyze the mechanism of the enhanced reduction of the thermal load in the bonded crystals, numerical simulations with a finite-element method were also performed.