Influence of cellular incorporation of n-3 eicosapentaenoic acid on intracellular Ca2+ concentration and membrane potential in vascular smooth muscle cells

Recombinant antibody fragments expressed in the cytoplasm of cells have considerable practical potential. However in the reducing environment of the cytoplasm, the intradomain disulphide bonds are not formed and the fragments are unstable and expressed in low yields. Here we attempted to overcome these limitations. We first isolated an antibody single chain Fv fragment that binds and activates an inactive mutant beta-galactosidase. We then subjected the gene encoding the scFv fragment to random mutation in vitro by error-prone polymerase chain reaction, and co-expressed the mutant beta-galactosidase and mutant antibody fragments in lac- bacteria. By plating on limiting lactose, we selected for antibody mutants with improved expression, and after four successive rounds of mutation and selection, isolated an antibody fragment that is expressed in the bacterial cytoplasm with yields of 0.5 g/l in a shaker flask (A600 nm of 5.5) and 3.1 g/l (A600 nm=33) in a fermentor. Analysis of the mutant antibody fragments revealed that the disulphide bonds are reduced in the cytoplasm, and that the fragments could be denatured and renatured efficiently under reducing conditions in vitro. This shows that with a suitable method of screening or selection, it is possible to make folded and functional antibody fragments in excellent yield in the cytoplasm.     

Radioactivity confinement capability of “PARC” concept for the future reprocessing

Aiming at TRU waste arising reduction and economical competitiveness for the future reprocessing, is proposed an advanced process concept which is named PARC (Partitioning Conundrum Key) process. Enhancement of confinement capability for long-lived nuclides in a simplified Purex process is the primary subject of this R&D project. Technologies for long-lived nuclide recovery are under development, focused on ¹⁴C and ¹²⁹I in head end, ²³⁷Np and ⁹⁹Tc in extraction, and ²⁴¹Am the daughter of ²⁴¹Pu in effluents. Those nuclides focused here are mobile in the environment and highly concerned as potential hazardous among the long-lived nuclides in spent fuels. New functions in PARC process concept are designed to mitigate the environmental impacts of reprocessing wastes and also to improve economy of reprocessing in the future.     

Mercury emissions from a coal-fired power plant in Japan

The emissions study for mercury was conducted at a 700 MW coal-fired plant for the combustion of three types of coal with mercury concentrations of 0.0063, 0.0367 and 0.065 mg/kg. The power plant is equipped with a cold-side electrostatic precipitator and wet type flue gas desulfurization system. During full load operation of the boilers, samples of the input and output streams such as coal, coal ash, ESP ash and post-ESP particulates and flue gas were collected. The Hg concentrations in solid were measured by cold-vapor atomic absorption spectrometry (AAS) after appropriate preparation and acid digestion. Gaseous Hg was collected using a mixed solution of potassium permanganate and sulfuric acid and the Hg concentrations in the samples were measured using cold-vapor AAS. The results were used to examine: (1) overall mass balances; (2) relative distribution in the power plant; (3) equilibrium of Hg species using MALT-2 calculation program; and (4) Hg concentrations in stack emissions. The mass balances estimated in this study were 100, 138 and 89%, respectively, for the coals. Total Hg concentrations in stack gas were 1.113, 0.422 and 0.712 microg(m3N), respectively, for the coals. More than 99.5% of the Hg in the stack emissions were in gaseous form and the proportion in particulate form was extremely low. The relative distribution of Hg in ESP, FGD and Stack ranged from 8.3 to 55.2%, 13.3 to 69.2% and 12.2% to 44.4%, respectively. The results indicated that factors other than the Hg concentration of coals and efficiency of pollution control devices might affect Hg emissions from coal-fired plant. The calculated equilibrium of the distribution of Hg species using the MALT2 program suggest that it is necessary to consider condensation mechanism to interpret the affect of Hg species on the variations of the removal efficiencies of Hg in the ESP.     

Some Considerations for Multiple-Unit Redundant Systems with Generalized Repair Time Distributions

The mean time to system failure (MTSF) for a 2-out-of-n system with repair is intuitively derived. The improvement factor, the ratio of the MTSF for a 2 out of n system with repair to that system without repair, is evaluated and its asymptotic behavior is studied.     

Glycerin-Induced Conformational Changes in Bombyx mori Silk Fibroin Film Monitored by 13C CP/MAS NMR and 1H DQMAS NMR

In order to improve the stiff and brittle characteristics of pure Bombyx mori (B. mori) silk fibroin (SF) film in the dry state, glycerin (Glyc) has been used as a plasticizer. However, there have been very limited studies on the structural characterization of the Glyc-blended SF film. In this study, ¹³C Cross Polarization/Magic Angle Spinning nuclear magnetic resonance (CP/MAS NMR) was used to monitor the conformational changes in the films by changing the Glyc concentration. The presence of only 5 wt % Glyc in the film induced a significant conformational change in SF where Silk I* (repeated type II β-turn and no α-helix) newly appeared. Upon further increase in Glyc concentration, the percentage of Silk I* increased linearly up to 9 wt % Glyc and then tended to be almost constant (30%). This value (30%) was the same as the fraction of Ala residue within the Silk I* form out of all Ala residues of SF present in B. mori mature silkworm. The ¹H DQMAS NMR spectra of Glyc-blended SF films confirmed the appearance of Silk I* in the Glyc-blended SF film. A structural model of Glyc-SF complex including the Silk I* form was proposed with the guidance of the Molecular Dynamics (MD) simulation using ¹H–¹H distance constraints obtained from the ¹H Double-Quantum Magic Angle Spinning (DQMAS) NMR spectra.     

Spreadsheet simulation of one-dimensional unsteady-state nonisothermal moisture diffusion in wood in the transverse direction

A spreadsheet for the numerical simulation of one-dimensional unsteady-state nonisothermal moisture diffusion in solid wood in the transverse direction below the fiber saturation point was developed in this work. Calculations for solving a pair of partial differential equations describing the moisture diffusion based on a finite-difference method were performed. Experimental validations revealed that the spreadsheet provides predictions of moisture diffusion in wood as accurately as a commercial simulation software package does. This spreadsheet simulation can help people involved in manufacturing, utilizing, or researching wood products to implement a simple and convenient numerical simulation of moisture diffusion in wood without any special skill and any specific software.     

Preparation and characterization of multilayered hydroxyapatite/silk fibroin film

We prepared multilayered films consisting of silk fibroin (SF) and hydroxyapatite (HAp) by alternating lamination using untreated SF and HAp-deposited SF films. Untreated SF films were prepared from a regenerated SF solution by air drying. HAp-deposited SF films were prepared by soaking methanol-treated SF films containing >5 wt% CaCl2 in a simulated body fluid with the ion concentration 1.5-fold higher than that of the standard one. The multilayered HAp/SF films had HAp layers with approximate thicknesses of 3-5 microm and SF layers with thicknesses of 40-70 microm. The bonding strength between the SF and HAp layers was significantly affected by temperature and compression time under the lamination method. The optimal conditions for achieving the maximum T-peel strength and beta-sheet contents were determined to be 130 degrees C for 4 min. The Young's modulus of the multilayered films (133.4 MPa) was higher than that of the films consisting of SF alone (92.5 MPa) under swollen conditions. The biocompatibility of the HAp-deposited SF films was analyzed by culturing of osteoblasts (MC3T3-E1) on a film. The results indicate that HAp-deposited SF films and SF films show similar degrees of cell adhesion and alkaline phosphatase activities.