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.     

Dissolution of Ti wires in sulphuric acid and hydrochloric acid solutions

Ti wire electrodes were immersed in acidic solutions containing H₂SO₄ and HCl of various concentrations at 353 K to evaluate corrosion rate by measurement of electric resistance change (resistometry). Addition of hydrochloric acid to sulphuric acid solution promoted depassivation of Ti. After depassivation, the immersion potential dropped to the hydrogen evolution potential and a hydride layer was formed on the surface. The hydride layer dissolved continuously in the acidic solution. SEM observation showed that Ti wires dissolved almost uniformly in the early stage and that the dissolution then trace became irregular due to nonuniform growth of the hydride layer. Dissolution rate of a Ti wire was estimated almost accurately by resistometry.     

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.     

Augmenting Edge-Connectivity between Vertex Subsets

Given a directed or undirected graph G=(V,E), a collection ${\mathcal{R}}=\{(S_{i},T_{i}) \mid i=1,2,\ldots,|{\mathcal{R}}|, S_{i},T_{i} \subseteq V, S_{i} \cap T_{i} =\emptyset\}$ of two disjoint subsets of V, and a requirement function $r: {\mathcal{R}} \to\mathbb{R}_{+}$ , we consider the problem (called area-to-area edge-connectivity augmentation problem) of augmenting G by a smallest number of new edges so that the resulting graph $\hat{G}$ satisfies $d_{\hat{G}}(X)\geq r(S,T)$ for all X?V, $(S,T) \in{\mathcal{R}}$ with S?X?V?T, where d _G (X) denotes the degree of a vertex set X in G. This problem can be regarded as a natural generalization of the global, local, and node-to-area edge-connectivity augmentation problems. In this paper, we show that there exists a constant c such that the problem is inapproximable within a ratio of $c\log{|{\mathcal{R}}|}$ , unless P=NP, even restricted to the directed global node-to-area edge-connectivity augmentation or undirected local node-to-area edge-connectivity augmentation. We also provide an ${\mathrm{O}}(\log{|{\mathcal{R}}|})$ -approximation algorithm for the area-to-area edge-connectivity augmentation problem, which is a natural extension of Kortsarz and Nutov’s algorithm (Kortsarz and Nutov, J. Comput. Syst. Sci., 74:662–670, 2008). This together with the negative result implies that the problem is ${\varTheta}(\log{|{\mathcal{R}}|})$ -approximable, unless P=NP, which solves open problems for node-to-area edge-connectivity augmentation in Ishii et al. (Algorithmica, 56:413–436, 2010), Ishii and Hagiwara (Discrete Appl. Math., 154:2307–2329, 2006), Miwa and Ito (J. Oper. Res. Soc. Jpn., 47:224–243, 2004). Furthermore, we characterize the node-to-area and area-to-area edge-connectivity augmentation problems as the augmentation problems with modulotone and k-modulotone functions.     

Preparation of modified poly(vinyl chloride) containing N,N-di(β-hydroxyethyl)dithiocarbamate and its reaction with metal ions

Reaction of N,N-di(β-hydroxyethyl)dithiocarbamate ion with poly(vinyl chloride) (PVC) was undertaken, and the reaction with metal ions of the polymer obtained was investigated. The effect of γ-irradiation on the reaction with metal ion was also studied. The modified PVC (PHDC) obtained from the reaction with N,N-di(β-hydroxyethyl)dithiocarbamate ion is pale yellow even after reaction at 100°C for 5 hr in dimethylformamide (DMF); it is soluble in dipolar solvents and its chlorine content is decreased considerably. This polymer reacted well with acetate salts of copper(II), nickel(II), zinc(II), and silver(I) heterogeneously in aqueous solution because of the introduction of hydrophilic groups (two hydroxy groups). The reactivity of the metal ions toward the polymer was of the order Ag(I) ? Cu(II) > Ni(II) > Zn(II). From the result of the reaction of γ-irradiated polymer with cupric ion, the polymer was judged to have fairly good antiradiation property.     

Effects of Pre-Treatment of a Silica-Supported Gallium Oxide Catalyst with H2 on Its Catalytic Performance for Dehydrogenation of Propane

The pre-treatment of a silica-supported gallium oxide catalyst with H₂ at 823 K increased the yield of aromatics and the selectivity to aromatics in the dehydrogenation of propane over the catalyst at 823 K. Gallium oxide in the catalyst was partially reduced with H₂ at 823 K. NH₃ desorption and DRIFTS studies on the gallium oxide catalyst suggest that the dehydrogenation of propane over a silica-supported gallium oxide catalyst would proceed in the following way: (1) the dehydrogenation of propane to produce propene would occur on Ga sites including Ga^δ+–H sites and (2) the aromatization of propene to aromatics on Ga–O–H acid sites.     

Synthesis of Super-Microporous Aluminosilicate Having Excellent Water Vapor Adsorption Property as an Adsorbent for an Adsorption Heat-Pump

The synthesis of super-microporous aluminosilicate with a pore diameter of 14.6 Å has been achieved, by means of synthesis under specified conditions with a skeletal material comprising TMOS and a small quantity of sodium aluminate, and a conventional alkyl-trimethylammonium halide (C₁₀TMABr) as a template. This super-microporous aluminosilicate has an adsorption capacity of 0.2 ml/g at a relative water vapor pressure of 0.2, and thus is promising as an adsorbent for an adsorption heat-pump.     

Thraustochytrid as a potential source of carotenoids

Thraustochytrids, marine protists whose dominant genera are Thraustochytrium and Schizochytrium, belong to the kingdom Chromista and are known as an industrial source of DHA. We describe here that thraustochytrid strain KH105, isolated as a DHA producer, also accumulates significant levels of β-carotene and xanthophylls including canthaxanthin and astaxanthin. A4-d cultivation using a medium composed of 10% glucose and less than 0.3% of nitrogen sources in a half-concentration of seawater gave an astaxanthin production up to 6.1 mg/L, and canthaxanthin content reached more than 10 mg/L under conditions where a higher concentration of nitrogen sources (6%) was employed. It might be advantageous in mass production systems for these carotenoids to be extracted readily by simply suspending the cells with organic solvents such as acetone and chloroform. Analyses on the morphological and life history features of the KH105 strain revealed that it belongs to the genus Schizochytrium. This particular species of thraustochytrids is thus considered to be a promising source of xanthophylls as well as DHA for use in the food industry.