Developmental ability of cloned embryos from neural stem cells

The success rate is generally higher when cloning mice from embryonic stem (ES) cell nuclei than from somatic cell nuclei, suggesting that the embryonic nature or the undifferentiated state of the donor cell increases cloning efficiency. We assessed the developmental ability of cloned embryos derived from cultured neural stem cell (NSC) nuclei and compared the success rate with that of embryos cloned from other donor cells such as differentiated NSCs, cumulus cells, Sertoli cells and ES cells in the mouse. The transfer of two-cell cloned embryos derived from cultured NSC nuclei into surrogate mothers produced five live cloned mice. However, the success rate (0.5%) was higher in embryos cloned from cultured NSC nuclei than from differentiated NSCs (0%), but lower than that obtained by cloning mice from other cell nuclei (2.2-3.5%). Although the in vitro developmental potential to the two-cell stage of the cloned embryos derived from NSC nuclei (73%) was similar to that of the cloned embryos derived from other somatic cell nuclei (e.g., 85% in Sertoli cells and 75% in cumulus cells), the developmental rate to the morula-blastocyst stage was only 7%. This rate is remarkably lower than that produced from other somatic cells (e.g., 50% in Sertoli cells and 54% in cumulus cells). These results indicate that the undifferentiated state of neural cells does not enhance the cloning efficiency in mice and that the arrest point for in vitro development of cloned embryos depends on the donor cell type.     

Synergistic Extraction of Am(III) and Eu(III) by Tris(2‐pyridylmethyl)Amine with Various Anions in 1,2‐Dichloroethane

The extraction equilibria of Am(III) and Eu(III) by using a tripodal ligand, tris(2‐pyridylmethyl)amine (tpa), with various lipophilic anions have been investigated. The extractability of both Am(III) and Eu(III) was increased by the combination of tpa and counteranions due to a synergistic effect. The separation factors between Am(III) and Eu(III) were also increased from 7.6 to 49 by the combination of counteranions and organic solvents. The extraction equilibria of Am(III) and Eu(III) with tpa in 1,2‐dichloroethane were determined by slope analysis. It was found that three anions and one molecule of the ligand coordinated to Am(III) and Eu(III) was extracted regardless of the anions.     

Determination of acrylamide in processed foods by LC/MS using column switching

An LC/MS method was developed for the determination of acrylamide (AA) in processed or cooked foods. AA was extracted with a mixture of water and acetone from homogenized food samples after the addition of 13C-labeled acrylamide (AA-1-(13)C) as an internal standard. The extract was concentrated, washed with dichloromethane for defatting, and cleaned up on Bond Elut C18, PSA and ACCUCAT cartridge-columns, and then AA was determined by LC/MS in the selected ion recording (SIR) mode. For the LC/MS analysis, four LC columns were connected in-line and the flow of the mobile phase was switched according to a time-program. Monitoring ions for AA were m/z 72 and 55, and those for AA-1-(13)C were m/z 73 and 56. AA and AA-1-(13)C were determined without interference from the matrices in all samples. The recoveries of AA from potato chips, corn snack, pretzel and roasted tea spiked at the level of 500 ng/g of AA were 99.5-101.0% with standard deviations (SD) in the range from 0.3 to 1.6%. The limits of detection and quantification of the developed method were 9 and 30 ng/g for AA in samples, respectively. The method was applied to the analysis of AA in various processed or cooked food samples purchased from retail markets. High levels of AA were found in potato chips and French-fried potato (467-3,544 ng/g). Fried and sugar-coated dough cakes (karinto) contained 374 and 1,895 ng/g. Corn snacks contained 117-535 ng/g of AA. Roasted foods (such as roasted sesame seed, roasted barley (mugi-cha), roasted tea (hoji-cha), coffee beans and curry powder) contained 116-567 ng/g of AA. Foods made from fish, egg and meat contained lower levels of AA than the plant-based foods. Foods containing much water showed a tendency to have low levels of AA compared with dry foods. The proposed method was applicable to the analysis of AA in variety of processed foods.     

Electron beam sterilization of cyclo olefin polymer leads to polymer degradation and production of alkyl radicals

We investigated the effects of electron‐beam (EB) sterilization on syringe barrels manufactured from cyclo olefin polymer (COP). The chemical structure of the polymer was determined by interpreting the ¹³C NMR and DEPT‐135 spectra of the COP resin. The antioxidants in the resin were identified by analyzing the liquid chromatography‐photo diode array‐mass spectrometry (LC‐PDA‐MS) data for the methanol extract of the resin and the gas chromatography‐mass spectrometry (GC‐MS) data for the supercritical methanol degradation products of the extract. NMR and LC‐PDA‐MS analyses revealed that EB sterilization produces degradation products in the COP main chain and reduces the quantity of the antioxidants in the COP resin. ESR spectra of the EB‐sterilized syringe barrels indicated the presence and location of alkyl radicals, which were generated in the COP main chain by EB sterilization. ESR analyses also indicated that the quantity of alkyl radicals decreased over time. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43498.     

Experimental investigation of melt pool behaviour during selective laser melting by high speed imaging

Melt pool behaviour and the characteristics of the surrounding metal powder during selective laser melting (SLM) are captured using high-speed imaging, and the influence of the substrate temperature and the line building sequence on the formation of the built structure and the generation of spatter particles is experimentally investigated. The results indicate that the substrate temperature has a measurable effect on the formation of droplets and the generation of spatter particles, while the volume specific energy density of the process was the principal factor affecting the built structure.     

Study of crater formation and sputtering process with large gas cluster impact by molecular dynamics simulations

Molecular dynamics (MD) simulations of large argon clusters impacting on silicon solid targets were performed in order to study the transient process of crater formation and sputtering. The MD simulations demonstrate that the initial momentum of incident cluster is transferred to target surface atoms through multiple collision mechanism, where the initial momentum, which is along to the surface normal before impact, is deflected to lateral direction. This momentum transfer process was analyzed by the calculation of the velocity at the crater edge (the interface between cluster and target). In the case of Ar₁₀₀₀ cluster impact on Si(1 0 0) target at low energy per atom less than 40 eV/atom, the maximum value of lateral velocity of the crater edge increases in proportional to the velocity of incident cluster atoms. On the other hand, the crater edge velocity saturates over 40 eV/atom of incident energy per atom. In this case, the whole of constituent cluster atoms are implanted into the target and expand in both lateral and reflective directions at the subsurface region of the target. These MD simulations demonstrated that this collisional process result in the high yield sputtering of the target atoms.     

Applications of microstructures fabricated by proton beam writing to electric-micro filters

Fabrication of high-aspect-ratio microstructures was performed by proton beam writing (PBW) using a microbeam line at Takasaki Ion Accelerators for Advanced Radiation Application (TIARA), JAEA Takasaki, JAPAN. As one of the applications of the high-aspect-ratio structures micro-machined by PBW, we utilized the high-aspect pillars for electric-micro filters of microbes such as Escherichia coli and Yeast based on the dielectrophoretic force. The filter is equipped with high-aspect pillars with a height of ～20 μm and a diameter of ～1 μm on a glass plate. Evaluation of the dielectrophoresis (DEP) device for capturing E. coli and Yeast was made using either observation by optical microscope or photoluminescence (PL) measurements.     

Detail analysis of fusion neutronics benchmark experiment for iron

In order to specify the best nuclear data on iron, the fusion neutronics benchmark experiment on iron at Japan Atomic Energy Agency (JAEA)/Fusion Neutronics Source (FNS) was analyzed in detail with MCNP-4C and the latest nuclear data libraries, JENDL-3.3, FENDL-2.1, JEFF-3.1 and ENDF/B-VII.0. As a result, totally the calculation result with ENDF/B-VII.0 agreed with the measurement best, except that it underestimated the measured neutron flux above 10 MeV with the depth. It was noted that the calculation result with JENDL-3.3 overestimated the measured neutrons below a few keV. Through the DORT calculations based on the iron data in ENDF/B-VII.0, it was found out that the first inelastic scattering cross-section data of ⁵⁷Fe in JENDL-3.3 caused the overestimation.     

Mouse Gastric Epithelial Cells Resist CagA Delivery by the Helicobacter pylori Type IV Secretion System

The initial step in bacterial infection is adherence of the bacterium to the target cell surface. Helicobacter pylori exploits the interaction of bacterial adhesin protein HopQ with human epithelial CEACAMs (CEACAM1, 5, and 6) to stably adhere to gastric epithelial cells, which is necessary for delivery of the H. pylori CagA oncoprotein into the epithelial cells via a type IV secretion system. In contrast to human CEACAMs, however, HopQ does not interact with Ceacam1 (mouse CEACAM1) in vitro or in CHO cells ectopically expressing Ceacam1. Since the mouse genome lacks Ceacam5 and Ceacam6, no significant HopQ–Ceacam interaction may occur in mouse gastric epithelial cells. Here, we found that the mouse stomach has a much lower expression level of Ceacam1 than the expression level of CEACAM1 in the human stomach. Consistently, mouse gastric epithelial cells resist CagA delivery by cagA-positive H. pylori, and the delivery is restored by ectopic expression of human CEACAM1 or CEACAM5 in mouse gastric epithelial cells. Thus, despite the fact that mice are routinely used for H. pylori infection studies, a low expression level of Ceacam1 in the mouse stomach together with the loss or greatly reduced interaction of HopQ with Ceacams make the mouse an inappropriate model for studying the role of H. pylori-delivered CagA in gastric pathogenesis, including the development of gastric cancer.