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.     

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.     

Visible-light-active titanium oxide photocatalyst realized by an oxygen-deficient structure and by nitrogen doping

A visible-light (vis)-active titanium oxide photocatalyst was prepared by a simple wet process: the calcination of the hydrolysis product of Ti(SO₄)₂ with ammonia using an ordinary electric furnace in dry air at 400 °C. The color of this photocatalyst was vivid yellow and absorbed light in the blue (400 nm) to bluish-green (550 nm) region exclusively. Its structure was characterized to anatase with oxygen-deficient stoichiometry by XRD and X-ray photoelectron spectroscopy. Nitrogen was also detected, but only in trace amounts. Using blue-light-emitting diodes as a light source, 540 ppm of acetone was decomposed within 36 h and a stoichiometric yield of CO₂ was obtained. From the results of the crystalline size D, it was found that vis-activity could be realized on polycrystalline particles and the grain-boundaries (GBs) are thought to be important, since oxygen vacancies are easily created in GBs, which could form a GB state. Finally, we concluded that oxygen-deficient sites formed in GBs are important to emerge vis-activity, and nitrogen doped in the part of oxygen-deficient sites are important as a blocker for reoxidation.     

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.

     

Isolation and characterization of a novel human pancreas-specific gene, pancpin, that is down-regulated in pancreatic cancer cells

We reviewed 19 children and adolescents with cervical spine congenital synostosis as in Klippel-Feil syndrome (KFS), with an average follow-up of 12.5 years. We paid particular attention to neurologic complications associated with cervical spine abnormalities. Five patients were affected by neurologic complications; four underwent a surgical procedure; and 14 had no neurologic finding. Two had hypermobility at one level, and one had hypermobility at two levels. We found that the more numerous the occipito-C1 abnormalities, the more significant the neurologic risk. In contrast, this risk was not related to the number of "mobile blocks" or to age. Various mechanisms of neural complications have been studied in the literature: medullary abnormality, spinal instability, narrowing of the cervical canal, and vascular dysfunction. Surgery is usually thought to be required in cases with neurologic complications. The indication for surgery is, however, less clear in cases of pure instability without neurologic involvement because surgery is likely to increase the future risks at mobile disks either above or below the fuse level. Careful clinical and radiologic observation is necessary in such patients. Magnetic resonance imaging (MRI) with lateral views in flexion and extension seem to be the best method for detecting impingement of the spine on the cord.     

Scale free interval graphs

Scale free graphs have attracted attention by their non-uniform structure that can be used as a model for various social and physical networks. In this paper, we propose a natural and simple random model for generating scale free interval graphs. The model generates a set of intervals randomly under a certain distribution, which defines a random interval graph. The main advantage of the model is its simpleness. The structure/properties of generated graphs are analyzable by relatively simple probabilistic and/or combinatorial arguments, which is different from many other models. Based on such arguments, we show for our random interval graph that its degree distribution follows a power law, and that it has a large average clustering coefficient.     

Quantum Monte Carlo Device Simulation of Nano-Scaled SOI-MOSFETs

Quantum transport properties of nano-scaled SOI-MOSFETs are investigated based on a quantum Monte Carlo (MC) device simulation. The quantum mechanical effects are incorporated in terms of a quantum correction of potential in the well-developed particle MC computational techniques. The ellipsoidal multi-valleys of silicon conduction band are also considered in the simulation. First, the validity of the quantum MC technique is verified by comparing the simulated results with a self-consistent Schrödinger-Poisson solution at thermal equilibrium. Then, we apply the technique to non-equilibrium and quasi-ballistic quantum transport in nano-scaled SOI-MOSFETs.     

Hydrolysis of beta-glucosyl ester linkage of p-hydroxybenzoyl beta-D-glucose, a chemically synthesized glucoside, by beta-glucosidases

To investigate the hydrolysis of glucosyl esters by beta-glucosidase, p-hydroxybenzoyl beta-D-glucose (pHBG) was chemically synthesized. The hydrolytic activity of some beta-glucosidases for pHBG was compared to that for p-nitrophenyl beta-glucoside (pNPG). The Clavibacter michiganense and Flavobacterium johnsonae enzymes could hydrolyze pHBG and steviol glycosides which are natural glucosyl esters. The commercial beta-glucosidase originating from Caldocellum saccharolyticum also hydrolyzes pHBG despite having no activity for steviol glycosides. The beta-glucosidase from Aspergillus niger cleaved the glucosyl ester linkage much more weakly than the glucosidic linkage. The pH-activity profile for the hydrolysis of pHBG was similar to that of pNPG by the C. saccharolyticum beta-glucosidase. The similar profiles for these substrates suggested that the active site for the glucosyl ester chemically resembles that for glucoside with respect to catalysis. Kinetic analysis of the C. saccharolyticum beta-glucosidase for mixed substrates of pHBG and pNPG showed that the hydrolysis of pHBG competed with that of pNPG. This result indicated that there is only one active site for both the glucosyl ester and glucoside. Mass spectroscopic analysis of the hydrolysates of pHBG in H218O suggested that beta-glucosidase hydrolyzes glucosyl esters between the anomeric carbon of glucose and the carbonyl oxygen, not between the carbonyl carbon and the carbonyl oxygen.