Metalation states versus enzyme activities of Cu, Zn-superoxide dismutase probed by electrospray ionization mass spectrometry

Cu, Zn-superoxide dismutase (SOD-1), an enzyme that catalyzes the disproportionation reaction of superoxide to produce oxygen and hydrogen peroxide, thereby protecting cells from oxidative stress, is a homodimer that coordinates one copper and one zinc ion per monomer. Cu (2+) and Zn (2+) ions play important roles in enzyme activity and structural stability, respectively. In addition, dimer formation is also essential for fulfilling the function of SOD-1. We here report on the reconstitution and enzyme activities of several metalation states of SOD-1 (Cu 4-, Cu 3Zn-, and Cu 2Zn 2-homodimers). Each metalation state of the reconstituted SOD-1 could be unambiguously differentiated by electrospray ionization mass spectrometry, the metal ions of which had been completely replaced by 99 atom % (63)Cu and (68)Zn stable isotopes. It was found that (1) the Cu 4-dimer possessed 84% of the activity of the native enzyme, (2) the Cu-site resisted being coordinated with Zn (2+) ions while the Zn-site could be bound with Cu (2+) ions, and (3) the simultaneous addition of the Cu (2+) and Zn (2+) ions to generate a fully metalated form produced the multiply metalated SOD-1 (Cu 4-, Cu 3Zn-, and Cu 2Zn 2-dimers), which were clearly distinguishable from one another by the use of the stable isotopes, while the sequential addition of Zn (2+) followed by the Cu (2+) ion predominantly produced a Cu 2Zn 2-dimer comparable to the native enzyme.     

Effects of carbon source on denitrification efficiency and microbial community structure in a saline wastewater treatment process

Two different denitrifying reactors were monitored in order to evaluate the effects of carbon source on denitrification efficiency and microbial community structure under various saline conditions. Nitrogen removal performances were determined when salinity concentrations increase gradually in acetate- or methanol-fed denitrifying reactor. As a result, acetate-fed process attained high nitrate removal at 0-10% NaCl, while methanol was proven beneficial electron donors at 0-3% NaCl. A parallel analysis of T-RFLP and cloning in the acetate-fed sludge showed that a specialized microbial population (i.e. the genera Halomonas and Marinobacter) adapted to a high saline environment. Meanwhile, there were no major changes of bacterial populations in the methanol-fed reactor at 4% NaCl, although the relative abundances of the genera Azoarcus and Methylophaga increased when salinity concentration was at 1-3% NaCl, indicating that methanol-utilizing populations in activated sludge was unable to adapt to a high saline environments (>4% NaCl).     

Residual stress measurement of Al/steel dissimilar friction stir weld

ABSTRACT

The dissimilar welds between aluminium (Al) alloy, A6061-T6, and stainless steel, type 304, were fabricated by a friction stir welding (FSW) technique. The FSW tool was offset to Al side and the probe was inserted only into Al plate. The softening occurred in Al side due to the heat input during FSW, while the hardness increased by the post-ageing treatment. Tensile strength of dissimilar weld also increased about 8.5% by the post-ageing. The residual stresses were measured based on the cos α method. The residual stresses parallel to the weld line, σ_xr, were predominantly tensile, while those perpendicular to the weld line, σ_yr, were compressive. Post-ageing treatment had little effect on the residual stresses.     

Conceptual design of spectrum changeable neutron calibration fields in JAERI/FRS

Spectrum changeable neutron calibration fields are planned to be established with an accelerator installed in Japan Atomic Energy Research Institute/Facility of Radiation Standards. The neutron fields are provided by bombarding a target surrounded by a moderator, with charged particles from the accelerator. In the fields, a wide variety of neutron spectra is provided with sufficient fluence rate for the calibration of dosemeters. In this study, necessity of the field was first discussed in view of relationship between readings of existing dosemeters and true dose equivalents where the dosemeters were used. Second, test simulation of neutron spectra was carried out with the Monte Carlo technique for some arrangements with a LiF target and quasi-cylindrical moderators with different materials. The simulated spectra were summarised in terms of fluence-average energy, fluence rate and calibration factor for the dosemeters.     

Development of the high-energy neutron fluence rate standard field in Japan with a peak energy of 45 MeV using the 7Li(p,n)7Be reaction at TIARA

In this study, we developed a 45 MeV neutron fluence rate standard of Japan. Quasi-monoenergetic neutrons with a peak energy of 45 MeV in the neutron standard field were produced by the ⁷Li(p,n)⁷Be reaction using a 50-MeV proton beam from an azimuthally varying field (AVF) cyclotron of the Takasaki Ion Accelerators for Advanced Radiation Application (TIARA). The neutron energy spectrum was measured using an organic liquid scintillation detector and a ⁶Li-glass scintillation detector by the time-of-flight method, and using a Bonner sphere spectrometer by the unfolding method. The absolute neutron fluence was determined using a proton recoil telescope (PRT) composed of the liquid scintillation detector and a Si(Li) detector that was newly developed in the present study. The detection efficiency of the PRT was obtained using the MCNPX code. The peak neutron production cross section for the ⁷Li(p,n)⁷Be reaction was also derived from the neutron fluence in order to confirm the neutron fluence of the TIARA high-energy neutron field. The peak neutron production cross section obtained in the present study was in good agreement with those of previous studies. The characteristics of the 45-MeV neutron field in TIARA were successfully evaluated in order to calibrate high-energy neutron detectors and high-energy neutron dosimeters.     

Thermoluminescence as a new research tool for the evaluation of geothermal activity of the Kakkonda geothermal system, northeast Japan

The thermoluminescence glow-curve of quartz in volcanic and pyroclastic rocks of the Miocene and Quaternary in the Kakkonda geothermal field was divided into L (low), M (medium) and H (high) peaks in order of increasing temperature. Thermoluminescence emission is independent of stratigraphic boundaries but it is closely related to surface geothermal manifestations. Thermally stimulated processes of thermoluminescence caused by natural annealing occurred in the Quaternary after the eruption of the Tamagawa Welded Tuffs; radiation storage processes then began, as a consequence of the temperature drop. Thermoluminescence behavior indicates natural temperature manifestations, together with the paleo-temperature history.The H peak was thermally stable compared to the L and M peaks, and the area within which the relative intensity of the H peak is less than 5% coincides with the surface zone where dominant fluid flow is convective. In addition, L and M peaks indicate that a relatively low-temperature fluid mixes with the hot upflow around the western margin of the ascending flow zone.Thermoluminescence characteristics reflect paleo-temperature history and are related to geothermal fluid flow. Thermoluminescence is an effective exploration technique for evaluating natural temperature manifestations and subterranean heat flow in geothermal systems.     

Study of cycle output improvement by work-fluid including phase change material

This paper deals with the output improvement of heating and cooling cycle by using the work-fluid including phase change material.The experimental study is carried out by heat exchange between work-fluid and heat transfer surface.The work-fluid is flown to a high temperature or a low temperature heat transfer surface from the narrow path.In order to increase the amount of the heat transmission,a trace of Diethylether(boiling point 34.8 ℃),as a phase change material(PCM),is added to the work-fluid.The parameters of the experiment are additive amount of PCM,the rotational speed of the displacer piston and the temperature of heat transfer surface.It is clarified that the increasing of engine cycle output is brought by the PCM addition.The effect of PCM addition is evaluated by output ratio which is defined from the experimental cycle output data.The requirements for acquiring the increasing effect of output by adding PCM are clarified.     

Evidence that up-regulation of the parotid Na(+)-K(+)-2Cl- cotransporter by hypertonic shrinkage is not duplicated by isotonic shrinkage

The application of Ca2+ mobilizing secretagogues to rat parotid acini results in a significant decrease in cell volume (15-30%) due to isotonic salt loss. It is often assumed that the effects of such an isotonic volume decrease can be mimicked by anisotonic cell shrinkage. We demonstrate that the Na(+)-K(+)-2Cl- cotransporter in these cells is up-regulated by Ca2+ mobilizing secretagogues as well as by cell shrinkage in hypertonic media. However, we find that although the protein kinase inhibitors staurosporine (0.3 microM) and K252a (0.6 microM) significantly blunt the latter up-regulation, they are without effect on the former. These observations suggest that hypertonic and isotonic shrinkage do not result in the activation of the same intracellular signalling pathways, and indicate that anisotonic volume perturbations may not provide good experimental models of physiologic isotonic volume changes.