Quantification of anaerobic ammonium-oxidizing bacteria in enrichment cultures by real-time PCR

The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a rotating disk reactor (RDR) biofilm in semi-batch cultures. Based on fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 36% ANAMMOX bacteria. Phylogenetic analysis revealed that all the detected clones were related to the previously reported ANAMMOX bacteria, Candidatus Brocadia anammoxidans (AF375994), with 92% sequence similarity. Furthermore, we successfully developed a real-time polymerase chain reaction (PCR) assay to quantify populations of ANAMMOX bacteria in the enrichment cultures. For this real-time PCR assay, PCR primer sets targeting 16S ribosomal RNA genes of ANAMMOX bacteria were designed and used. The quantification range of this assay was 6 orders of magnitude, from 8.9x10(1) to 8.9x10(6) copies per PCR, corresponding to the detection limit of 3.6x10(3) target copies mL(-1). A significant correlation was found between the increase in copy numbers of 16S rRNA gene of ANAMMOX bacteria and the increase in nitrogen removal rates in the enrichment cultures. Quantifying ANAMMOX bacterial populations in the enrichment culture made it possible to estimate the doubling time of the enriched ANAMMOX bacteria to be 3.6 to 5.4 days. The real-time PCR assay gave comparable population sizes in the enrichment cultures with the FISH results. These results suggest that the real-time PCR assay developed in this study is useful and reliable for quantifying the populations of ANAMMOX bacteria in environmental and engineering samples.     

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.     

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.     

Present Status of the Nd:YAG Thomson Scattering System Development for Time Evolution Measurement of Plasma Profile on Heliotron J

A new high repetition rate Nd:YAG Thomson scattering system is developed for the Heliotron J helical device.A main purpose of installing the new system is the temporal evolution measurement of a plasma profile for improved confinement physics such as the edge transport barrier (H-mode) or the internal transport barrier of the helical plasma.The system has 25 spatial points with ～10mm resolution.Two high repetition Nd:YAG lasers (>550mJ@ 50Hz) realize the measurement of the time evolution of the plasma profile with ～10ms time intervals.Scattered light is collected by a large concave mirror (D=800 mm,f/2.25) with a solid angle of ～100 mstr and transferred to interference filter polychromators by optical fiber bundles in a staircase form.The signal is amplified by newly designed fast preamplifiers with DC and AC output,which reduces the low frequency background noise.The signals are digitized with a multi-event QDC,fast gated integrators.The data acquisition is performed by a VME-based system operated by the CINOS.     

The result of a wall failure in-pile experiment under the EAGLE project

The WF (wall failure) test of the EAGLE program, in which 2 kg of uranium dioxide fuel-pins were melted by nuclear heating, was successfully conducted in the IGR (Impulse Graphite Reactor) of NNC/Kazakhstan. In this test, a 3 mm-thick stainless steel (SS) wall structure was placed between fuel pins and a 10 mm-thick sodium-filled channel (sodium gap). During the transient, fuel pins were heated, which led to the formation of a fuel-steel mixture pool. Under the transient nuclear heating condition, the SS wall was strongly heated by the molten pool, leading to wall failure. The time needed for fuel penetration into the sodium-filled gap was very short (less than 1 s after the pool formation). The result suggests that molten core materials formed in hypothetical LMFBR core disruptive accidents have a certain potential to destroy SS-wall boundaries early in the accident phase, thereby providing fuel escape paths from the core region. The early establishment of such fuel escape paths is regarded as a favorable characteristic in eliminating the possibility of severe re-criticality events. A preliminary interpretation on the WF test results is presented in this paper.