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.     

The World robot summit disaster robotics category – achievements of the 2018 preliminary competition

The World Robot Summit is a robot Olympics and aims to be held in a different country every four years from 2020. The concept of the Plant Disaster Prevention challenge is daily inspections, checks, and emergency response in industrial plants, and in this competition, robots must carry out these types of missions in a mock-up plant. The concept of the Tunnel Disaster Response and Recovery challenge is emergency response to tunnel disasters, and is a simulation competition whereby teams compete to show their ability to deal with disasters, by collecting information and removing debris. The Standard Disaster Robotics challenge assesses, in the form of a contest, the standard performance levels of a robot that are necessary for disaster prevention and emergency response. The World Robot Summit Preliminary Competition was held at Tokyo Big Sight in October 2018, and 36 teams participated in the Disaster Robotics Category. UGVs and UAVs contended the merits of new technology for solving complex problems, using core technologies such as mobility, sensing, recognition, performing operations, human interface, autonomous intelligence etc., as well as system integration and implementation of strategies for completing missions, gaining high-level results.     

Aminoglycosides prevent and dissociate the aggregation of platelets in patients with EDTA-dependent pseudothrombocytopenia

Although EDTA-dependent pseudothrombocytopenia (EDTA-PTCP) is of practical importance because failure to recognize this clinical entity may result in misdiagnosis and subsequent mismanagement of the patients, the pathophysiological nature of EDTA-PTCP remains unknown. To develop an effective way to evaluate the platelet counts in patients with EDTA-PTCP, we introduced aminoglycosides-supplemented anticoagulating agents. When kanamycin was pre-supplemented with EDTA for anticoagulating blood samples from EDTA-PTCP patients there was no significant change in the platelet counts and the morphology of blood cells after 150 min of incubation at room temperature. Furthermore, when kanamycin was added to EDTA-anticoagulated blood samples from EDTA-PTCP patients within 30 min after blood withdrawal, rapid dissociation of platelets without apparent morphological changes of blood cells was observed, and complete blood cell counts as well as the histogram patterns were almost the same as those examined immediately after blood sampling. The dissociation of aggregated platelets was also detected when other antibiotics were used, although it was associated with some extent of morphological changes of blood cells. These findings indicate that the supplementation of aminoglycosides either before or after blood sampling is a useful method for the diagnosis EDTA-PTCP and for the evaluation of platelet counts in patients with EDTA-PTCP.     

Active sites of Cu-ZSM-5 for the decomposition of acrylonitrile

The catalytic decomposition of acrylonitrile (AN) over Cu-ZSM-5 prepared with various Cu loadings was investigated. AN conversion, during which the nitrogen atoms in AN were mainly converted to N₂, increased as Cu loading increased. N₂ selectivities as high as 90–95% were attained. X-ray diffraction measurements (XRD) and temperature-programmed reduction by H₂ (H₂-TPR) showed the existence of bulk CuO in Cu-ZSM-5 with a Cu loading of 6.4 wt% and the existence of highly dispersed CuO in Cu-ZSM-5 with a Cu loading of 3.3 wt%. Electron spin resonance measurements revealed that Cu-ZSM-5 contains three forms of isolated Cu²⁺ ions (square-planar, square-pyramidal, and distorted square-pyramidal). The H₂-TPR results suggested that in Cu-ZSM-5 with a Cu loading of 2.9 wt% and below, Cu⁺ existed even after oxidizing pretreatment. The activity of AN decomposition over Cu/SiO₂ suggested that CuO could form N₂, but, independent of the CuO dispersion, nitrogen oxides (NO_x) were formed above 350 °C. Cu⁺ and the square-pyramidal and distorted square-pyramidal forms of Cu²⁺ showed low activity for AN decomposition. Temperature-programmed desorption of NH₃ suggested that N₂ formation from NH₃ proceeded on Cu²⁺, resulting in the formation of Cu⁺. The Cu⁺ ions were oxidized to Cu²⁺ at around 300 °C. Thus, high N₂ selectivity over Cu-ZSM-5 with a wide range of temperature was probably attained by the reaction over the square-planar Cu²⁺, which can be reversibly reduced and oxidized.     

Electric field-assisted and field-depressed segregation of reactive metals to the bonding interface in braze alloy joining

Segregation of reactive metals at the bonding interface has been observed in various ceramic and/or metal joints bonded with reactive metal-bearing braze alloys. When a d.c. of 20 mAcm⁻² is applied to the ceramic/braze/ceramic system at a brazing temperature of, say, 1373 K, the electric field assists the segregation at the braze-ceramic interface on the cathode side and suppresses the segregation at the interface on the anode side. This may imply that reactive metal atoms in the braze can migrate as a cation. E.m.f. measurement on the ceramic (AIN or ZrB₂)-metal foil systems with increasing temperature shows that a negative e.m.f. to the ceramic pole appears from about 900 K for AIN and from 500 K for ZrB₂, as does the thermally stimulated current in polymers. These temperatures coincide well with those where the electrical conductivity of AIN and ZrB₂, respectively, begins to increase with increasing temperature. Therefore, it is considered that the polarization of the ceramics may take place and assist the migration, and consequently segregation, of reactive metals in braze alloys to the braze-ceramic interface during brazing.     

Structure and mechanical properties of polyethylene-fullerene composites

The microhardness of films of fullerene-polyethylene composites prepared by gelation from semidilute solution, using ultrahigh molecular weight polyethylene (PE) (6×10⁶), has been determined. The composite materials were characterized by optical microscopy and X-ray diffraction techniques. The microhardness of the films is shown to increase notably with the concentration of fullerene particles within the films. In addition, a substantial hardening of the composites is obtained after annealing the materials at high temperatures (T _a=130 °C) and long annealing times (t _a=10⁵s). The hardening of the composites with annealing temperature has been identified with the thickening of the PE crystalline lamellae. Comparison of X-ray scattering data and the microhardness values upon annealing leads to the conclusion of phase separation of C₆₀ molecules from the polyethylene crystals within the material. The temperature dependence is discussed in terms of the independent contribution of the PE matrix of the C₆₀ aggregates to the hardness value.     

Effect of surfactants on phenol removal by the method of polymerization and precipitation catalysed by Coprinus cinereus peroxidase

The removal of phenol by peroxidase‐catalysed polymerization was examined using Coprinus cinereus peroxidase in the presence of surfactants. The non‐ionic surfactants with poly(oxyethene) residues, Triton X‐100, Triton X‐405 and Tween 20, enhanced the phenol removal efficiency at a level similar to high relative molecular mass poly(ethylene glycol) (relative molecular mass 3000). Although the improvement in the removal efficiency was less than that of Triton X‐100, Span 20, sodium lauryl sulfate (SDS) and lauryl trimethylammonium bromide (DTAB) also enhanced the removal efficiency. The requirement of the enzyme for almost 100% removal of 100 mg dm^?3 phenol decreased to one‐fourth by the addition of 30 mg dm^?3 Triton X‐100. Triton X‐100, Triton X‐405, Tween 20 and DTAB could reactivate the enzyme precipitated with the phenol polymer, leading to the restarting of the phenol removal reaction. Copyright © 2003 Society of Chemical Industry     

0.5-μm 3.3-V BiCMOS standard cells with 32-kilobyte cache andten-port register file

BiCMOS standard cell macros, including a 0.5-W 3-ns register file, a 0.6-W 5-ns 32-kbyte cache, a 0.2-W 3-ns table look-aside buffer (TLB), and a 0.1-W 3-ns adder, are designed with a 0.5-μm BiCMOS technology. A supply voltage of 3.3 V is used to achieve low power consumption. Several BiCMOS/CMOS circuits, such as a self-aligned threshold inverter (SATI) sense amplifier and an ECL HIT logic are used to realize high-speed operation at the low supply voltage. The performance of the BiCMOS macros is verified using a fabricated test chip     

Subarachnoid haemorrhage induced proliferation of leptomeningeal cells and deposition of extracellular matrices in the arachnoid granulations and subarachnoid space. Immunhistochemical study

Subarachnoid haemorrhage (SAH) often leads to subarachnoid fibrosis and resultant normal pressure hydrocephalus; however, how subarachnoid fibrosis occurs is unknown. We examined the changes within arachnoid granulations (AGs) and the subarachnoid space (SAS) chronologically at the parasagittal region obtained from patients with SAH at autopsy and made comparison with controls by immunostaining for cytokeratin, specific marker for leptomeningeal cells and by the elastica Masson-Goldner methods. Within a week some AGs were torn, and many inflammatory cells filled the AGs and SAS. Cytokeratin positive cells were scarce. During the next two weeks cytokeratin positive cells increased. After three weeks, AGs and SAS were filled by dense deposits of extracellular matrices surrounded by multiple layers of leptomeningeal cells.     

Bias-stress-induced increase in parasitic resistance of InP-based InAlAs/InGaAs HEMTs

The reliability of InP-based HEMTs is studied, focussing on how it is affected by the doped layer material and gate recess structure. Bias-and-temperature stress tests reveal that fluorine-induced donor passivation in the recess region, formed adjacent to the gate electrode, causes the source resistance (R_s) to increase at large drain bias voltages. The increase in R_s can be prevented by using InP or InAlP as the carrier supply layer material instead of InAlAs. On the other hand, the increase in the drain resistance (R_d) does not depend on the material of the carrier supply layer, which suggests that a mechanism different from that in the case of R_s should be considered. It is also found that a deep gate recess suppresses the increase in R_d after long-term stressing.