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.     

Hinokitiol induces differentiation of teratocarcinoma F9 cells

Hinokitiol, a constituent of the wood of Chamaecyparis taiwanensis, was found to induce differentiation of teratocarcinoma F9 cells. When examined by the agar-overlay method, in which expression of plasminogen activator as a differentiation marker protein was detected, this compound exhibited a dose- and time-dependent induction. Induction of differentiation by hinokitiol occurred irreversibly and required its addition for more than 12h. Among its structure-related compounds tested, tropolone and two colchicine-related compounds exerted potent activities comparable to that of hinokitiol. These findings indicate that free tropolone structure in the molecules plays an essential role in inducing differentiation of F9 cells. Hinokitiol showed a strong inhibitory effect of DNA synthesis in very early stages of culture, suggesting that this effect may be responsible for triggering differentiation of F9 cells.     

Oxidation of Fe (II) in sulfuric acid solutions with dissolved molecular oxygen

The oxidation of Fe(II) with dissolved molecular oxygen was studied in sulfuric acid solutions containing 0.2 mol . dm^-3 FeSO₄ at temperatures ranging from 343 to 363 K. In solutions of sulfuric acid above 0.4 mol . dm^-3, the oxidation of Fe (II) was found to proceed through two parallel paths. In one path the reaction rate was proportional to both [Fe²⁺]² and po₂, exhibiting an activation energy of 51.6 . kJ mol^-1. In another path the reaction rate was proportional to [Fe²⁺]², [SO_4-], and po₂ with an activation energy of 144.6 kJ . mol^-1. A reaction mechanism in which the SO_4- ions play an important role was proposed for the oxidation of Fe(II). In dilute solutions of sulfuric acid below 0.4 mol . dm^-3, the rate of the oxidation reaction was found to be proportional to both [Fe(II)]² and Po₂, and was also affected by [H⁺] and [SO_2- ⁴]. The decrease in [H⁺] resulted in the increase of reaction rate. The discussion was further extended to the effect of Fe (III) on the oxidation reaction of Fe (II).     

A Study of Shadowing on Indoor Visible-Light Wireless Communication Utilizing Plural White LED Lightings

White LED is considered as a strong candidate for the future lighting technology. We have proposed an optical wireless communication system that employs white LEDs for indoor wireless networks. In this system, LED is used not only as a lighting device, but also as a communication device. The transmitter has large optical power and large emission characteristics to function as lighting device. And the system has specific wireless channel impulse response differing from infrared wireless communication. In this paper, we discuss about shadowing effect on the system utilizing plural LED lightings including the performance of ISI based on the impulse response. We consider the downlink transmission based on TDMA and evaluate the shadowing effect caused by pedestrians with computer simulation. When the shadowing often occurs at 800 Mb/s, the performance of outage call duration rate and blocking rate are improved by using 3 LED lightings compared with 1 or 2 LED lightings. And, we show that the system with the optimal number of the LED lighting is robust against shadowing and can accommodate more calls. Toshihiko Komine was born in Shizuoka, Japan, on November 17, 1978. He received the B.E. and M.E. degrees in Information and Computer Science from Keio University, Yokohama, Japan, in 2001 and 2003 respectively. He is currently studying for the Ph.D. degree at Department of Information and Computer Science, Keio University. His current research interests are optical wireless communications and LED communications. Shinichiro Haruyama is a professor at Department of Information and Computer Science, Faculty of Science and Technology, Keio University, Yokohama, Japan. He received an M.S. in engineering science from University of California at Berkeley in 1983 and a Ph.D. in computer science from the University of Texas at Austin in 1990. He worked for Bell Laboratories of AT{&}T and Lucent Technologies, U.S.A from 1991 to 1996, and for Sony Computer Science Laboratories, Inc. from 1998 to 2002. His research interests include reconfigurable system, system design automation, wireless communication, and visible light communication. Masao Nakagawa was born in Tokyo, Japan in 1946. He received the B.E., M.E. and Ph.D. degrees in electrical engineering from Keio University, Yokohama, Japan, in 1969, 1971 and 1974 respectively. Since 1973, he has been with the Department of Electrical Engineering, Keio University, where he is now a Professor. His research interests are in CDMA, consumer Communications, Mobile communications, ITS (Intelligent Transport Systems), Wireless Home Networks, and Visible light Communication. He received 1989 IEEE Consumer Electronics Society Paper Award, 1999-Fall Best Paper Award in IEEE VTC, IEICE Achievement Award in 2000, IEICE Fellow Award in 2001. He was the executive committee chairman on International Symposium on Spread Spectrum Techniques and Applications in 1992 and the technical program committee chairman of ISITA (International Symposium on Spread Spectrum Techniques and Applications) in 1994. He is an editor of Wireless Personal Communications and was a guest editor of the special issues on “CDMA Networks I, II, III and IV” published in IEEE JSAC in 1994 (I and II) and 1996 (III and IV). He chairs the Wireless Home Link sub-committee in MMAC (Multimedia Mobile Access Communication Promotion Committee).     

Suppression of photo-induced dilation in cyanide treated hydrogenated amorphous silicon films

Effects of cyanide (CN) treatment with hydrogenated amorphous silicon (a-Si:H) films have been investigated. The decrease of ΔV/V was observed in cyanide treated a-Si:H films and the successive thermal annealing at 200°C after CN treatment induced the further reduction of the ΔV/V. XPS spectra show the indirect evidence that the cyanide species is present within 10 nm from the hydrogenated amorphous silicon surface. The results of CN treatment with a-Si:H solar cells are demonstrated.     

Effects of intravenous lansoprazole on acute gastric mucosal lesions and acid secretion

The effects of lansoprazole given intravenously on gastric mucosal lesions, gastric bleeding and acid secretion were investigated in rats in comparison with those of omeprazole, famotidine and ranitidine. Lansoprazole inhibited the formation of gastric mucosal lesions in rats induced by water-immersion stress or aspirin with ID50 values of 0.26 and 0.99 mg/kg, respectively, and also inhibited gastric bleeding induced by hemorrhagic shock or water-immersion stress with ID50 values of 0.46 and 1.22 mg/kg, respectively. Lansoprazole was more potent than omeprazole, famotidine and ranitidine in inhibiting gastric mucosal lesions and hemorrhagic shock- or stress-induced bleeding. Famotidine and ranitidine showed negligible inhibition of water-immersion stress-induced gastric bleeding. Lansoprazole strongly inhibited water-immersion stress-stimulated acid secretion in rats, whereas famotidine and ranitidine did not show a potent inhibitory effect. These results indicate that lansoprazole exerts prominent inhibitory actions against the formation of gastric mucosal lesions and gastric bleeding by inhibiting acid secretion, and they show that it is superior to histamine H2-receptor antagonists in inhibiting stress-induced gastric bleeding.     

Multiobjective dynamic load-dispatching method for optimal operation of power systems

A new practical method is proposed which gives a quasi-optimal solution of the dynamic load-dispatching problem formulated by a multiconstraint multiobjective optimization problem. Here, the multiple constraints are rate reserve constraints and power flow ones, and the multiple objectives are fuel cost and CO₂ emission. The proposed method is an integration of the following three techniques: (1) “Group Dispatch Scheme,” which is developed by the authors, is used to satisfy a violated constraint easily. This scheme consists of three steps. The first step is to classify all generators into two groups. One is the group which should be more loaded to satisfy the violated constraint, and the other is the should-be less loaded group. The second step is to add some load to the former group, and to subtract the same load from the latter. The third step is to dispatch each group load to each groups generators, respectively; (2) the dynamic load-dispatching algorithm is used which provides, in practical time, quasi-optimal generation trajectories; (3) the conventional weighting parametric method is used to obtain a Pareto optimal solution of multiobjective problems. The effectiveness of the method is clarified by a computer simulation on the actual power system of Kansai Electric Power Company.     

Dose-dependent induction of both pepsinogen-altered pyloric glands and adenocarcinomas in the glandular stomach of C3H mice treated with N-methyl-N-nitrosourea

BACKGROUND: Cigarette smoking has been shown to increase oxidative DNA damage in human sperm cells. Assessment of the role of cigarette smoking in the etiology of childhood cancer has focused primarily on the effect of maternal smoking. Similar studies in relation to paternal smoking, however, have been inconclusive. Few studies have evaluated the effect of paternal smoking in the preconception period, and most of these could not disentangle the effects of paternal from maternal smoking. PURPOSE: We investigated the relationship of paternal smoking, particularly in the preconception period, with childhood cancer among offspring of the nonsmoking mothers. METHODS: We conducted a population-based, case-control study in Shanghai, People's Republic of China, where the prevalence of smoking is high among men but extremely low among women. The study included 642 childhood cancer case patients (<15 years of age) and their individually matched control subjects. Information concerning parental smoking, alcohol drinking, and other exposures of the index child was obtained by direct interview of both parents of the study subjects. Odds ratios (ORs), derived from conditional logistic regression models, were used to measure the association between paternal smoking and risk of childhood cancers. RESULTS AND CONCLUSIONS: Paternal preconception smoking was related to a significantly elevated risk of childhood cancers, particularly acute leukemia and lymphoma. The risks rose with increasing pack-years of paternal preconception smoking for acute lymphocytic leukemia (ALL) (P for trend = .01), lymphoma (P for trend = .07), and total cancer (P for trend = .006). Compared with children whose fathers had never smoked cigarettes, children whose fathers smoked more than five pack-years prior to their conception had adjusted ORs of 3.8 (95% confidence interval [CI] = 1.3-12.3) for ALL, 4.5 (95% CI = 1.2-16.8) for lymphoma, 2.7 (95% CI = 0.8-9.9) for brain tumors, and 1.7 (95% CI = 1.2-2.5) for all cancers combined. Statistically significant increased risks of cancer were restricted to children under the age of 5 years at diagnosis or those whose fathers had smoked during all of the 5 years prior to conception. IMPLICATIONS: Further studies are needed to confirm the association of paternal smoking with increased risk of cancer in offspring, to clarify the pattern of risks in relation to the timing of cigarette smoking, and to elucidate the biologic mechanism involved in predisposing the offspring to cancer. For example, it may be that paternal smoking induces prezygotic genetic damage that, in turn, acts as the predisposing factor.     

Flexural properties of moldings of rigid polyurethane made by reaction injection molding

Flexural properties of moldings made by Reaction Injection Molding (RIM), which are structural foams consisting of high density skin and low density core, were investigated by three-point bending tests. Two failure modes were observed in bending tests of the moldings made by RIM, and they are classified as follows according to the density ratio of skin layer to core layer: the opposite side of the skin layer to which load was subjected failed by tensile stress: and the same side of the skin layer to which load was subjected failed by compressive stress, causing wrinkling or buckling. Then the conventional composite beam theory was applied to the former failure mode and Hoff s buckling theory to the latter, and equations were derived to predict the flexural properties of the structural foams, which involved buckling from the flexural properties of solid construction. In addition, it has been shown that there exists a density distribution that maximizes the flexural strength of the moldings made by RIM with a given overall density. The results obtained here should be useful to the optimum structural design of moldings made by RIM.