Site‐specific Protein Cross‐Linking with Genetically Incorporated 3,4‐Dihydroxy‐L‐Phenylalanine

Come together right now with L ‐DOPA : Chemical cross‐linking is widely used to study protein–protein interactions. However, many cross‐linking agents suffer from low reactivity or selectivity. An efficient and selective reaction of site‐specific protein cross‐linking was achieved using genetically incorporated 3,4‐dihydroxy‐L ‐phenylalanine.

     

Synthesis of proton conductive membranes based on inorganic–organic hybrid structure bound with phosphonic acid

Proton conductive inorganic–organic hybrid membranes were synthesized from styrene derivatives of alkoxysilane and ethyl 2-[3-(dihydroxyphosphonyl)-2-oxopropyl] acrylate (EPA) through copolymerization followed by sol–gel reaction. Self-standing, homogeneous and transparent hybrid membranes with chemically bound phosphonic acid groups were synthesized. FT-IR analysis exhibited the hybrid membranes included phosphonic acid groups. ¹³C and ²⁹Si NMR studies indicated that alkoxysilyl functionalized styrene derivatives were not only copolymerized with EPA but also condensed yielding Si–O–Si linkages by sol–gel reaction. TG–DTA analysis revealed that these membranes were thermally stable up to 200 °C in dry O₂. The proton conductivities of the hybrid membranes increased with phosphonic acid content and temperature. The P/Si ratio of the membrane was dependent on the number of alkoxy group in the starting alkoxysilane. The hybrid membrane from (dimethylmethoxysilylmethyl)styrene (DMMSMS(M))/EPA = 1/6 revealed proton conductivities of 6.3 × 10⁻³ and 2.4 × 10⁻⁴ S cm⁻¹ at 68.0% relative humidity and 18.8% relative humidity, respectively, at 140 °C.     

High air stability of threshold voltage on gate bias stress in pentacene TFTs with a hydroxyl-free and amorphous fluoropolymer as gate insulators

We investigated the air stabilities of threshold voltages (V_th) on gate bias stress in pentacene thin-film transistors (TFTs) with a hydroxyl-free and amorphous fluoropolymer as gate insulators. The 40-nm-thick thin films of spin-coated fluoropolymer had excellent electrical insulating properties, and the pentacene TFTs exhibited negligible current hysteresis, low leakage current, a field-effect mobility of 0.45 cm²/Vs and an on/off current ratio of 3 × 10⁷ when it was operated at −20 V in ambient air. After a gate bias stress of 10⁴ s, a small V_th shift below 1.1 V was obtained despite non-passivation of the pentacene layer. We have discussed that the excellent air stability of V_th was attributed to the insulator surface without hydroxyl groups.     

Voltage holding study of 1 MeV accelerator for ITER neutral beam injector

Voltage holding test on MeV accelerator indicated that sustainable voltage was a half of that of ideal quasi-Rogowski electrode. It was suggested that the emission of the clumps is enhanced by a local electric field concentration, which leads to discharge initiation at lower voltage. To reduce the electric field concentration in the MeV accelerator, gaps between the grid supports were expanded and curvature radii at the support corners were increased. After the modifications, the accelerator succeeded in sustaining -1 MV in vacuum without beam acceleration. However, the beam energy was still limited at a level of 900 keV with a beam current density of 150 A∕m(2) (346 mA) where the 3 × 5 apertures were used. Measurement of the beam profile revealed that deflection of the H(-) ions was large and a part of the H(-) ions was intercepted at the acceleration grid. This causes high heat load on the grids and the breakdowns during beam acceleration. To suppress the direct interception, new grid system was designed with proper aperture displacement based on a 3D beam trajectory analysis. As the result, the beam deflection was compensated and the voltage holding during the beam acceleration was improved. Beam parameter of the MeV accelerator was increased to 980 keV, 185 A∕m(2) (427 mA), which is close to the requirement of ITER accelerator (1 MeV, 200 A∕m(2)).     

High-temperature tribological properties of spark-plasma-sintered Al2O3 composites containing barite-type structure sulfates

Al₂O₃–50BaSO₄–20Ag, Al₂O₃–50BaSO₄–10SiO₂, Al₂O₃–50(mass%)SrSO₄, Al₂O₃–50PbSO₄–5SiO₂, Al₂O₃–50BaSO₄ and Al₂O₃–50BaCrO₄ composites (mass%) were prepared by spark plasma sintering and their microstructure and high-temperature tribological properties were evaluated. Al₂O₃–50BaSO₄–20Ag composites (mass%) showed the lowest friction coefficients at the temperature ranging from 473 to 1073 K. Thin Ag film was observed on the wear tracks of the composites above 473 K. In addition, the friction coefficients of Al₂O₃ composites containing SrSO₄ and PbSO₄ were as low as those of Al₂O₃–BaSO₄ and Al₂O₃–BaCrO₄ composites at the temperatures up to 1073 K. The thin films formed on the wear tracks of the Al₂O₃–SrSO₄ composites were composed of Al₂O₃ and SrSO₄ phases, while the films formed on the wear tracks of the Al₂O₃–PbSO₄–SiO₂ composites consisted of Al₂O₃, PbSO₄ and SiO₂ phases.     

Automated Synthesis of (rac)‐, (R)‐, and (S)‐[18F]Epifluorohydrin and Their Application for Developing PET Radiotracers Containing a 3‐[18F]Fluoro‐2‐hydroxypropyl Moiety

To introduce the 3‐[¹⁸F]fluoro‐2‐hydroxypropyl moiety into positron emission tomography (PET) radiotracers, we performed automated synthesis of (rac)‐, (R)‐, and (S)‐[¹⁸F]epifluorohydrin ([¹⁸F] 1 ) by nucleophilic displacement of (rac)‐, (R)‐, or (S)‐glycidyl tosylate with ¹⁸F^? and purification by distillation. The ring‐opening reaction of (R)‐ or (S)‐[¹⁸F] 1 with phenol precursors gave enantioenriched [¹⁸F]fluoroalkylated products without racemisation. We then synthesised (rac)‐, (R)‐, and (S)‐ 2‐{5‐[4‐(3‐[¹⁸F]fluoro‐2‐hydroxypropoxy)phenyl]‐2‐oxobenzo[d]oxazol‐3(2H)‐yl}‐N‐methyl‐N‐phenylacetamide ([¹⁸F] 6 ) as novel radiotracers for the PET imaging of translocator protein (18 kDa) and showed that (R)‐ and (S)‐[¹⁸F] 6 had different radioactivity uptake in mouse bone and liver. Thus, (rac)‐, (R)‐, and (S)‐[¹⁸F] 1 are effective radiolabelling reagents and can be used to develop PET radiotracers by examining the effects of chirality on their in vitro binding affinities and in vivo behaviour.     

Development of “STORK”, a watermelon-harvesting robot

Recently, the production of heavy fruit and vegetables has been decreasing in Japan because strenuous labor is require to harvest them. A robot would allow them to be harvested more easily. We have developed the robot “STORK” to harvest watermelons. STORK has a low mass and a long working range. The position accuracy and repeatability of the manipulator, the required vacuum, and the allowance for position error of the vacuum pad were tested.     

Evidence that a phosphatidylinositol 3,4,5-trisphosphate-binding protein can function in nucleus

PIP3BP is a phosphatidylinositol 3,4,5-trisphosphate-binding protein (PIP3BP) abundant in brain, containing a zinc finger motif and two pleckstrin homology (PH) domains. Staining of rat brain cells with anti-PIP3BP antibody and determination of localization of PIP3BP fused to the green fluorescent protein (GFP-PIP3BP) revealed that PIP3BP was targeted to the nucleus. Targeting was dependent on a putative nuclear localization signal in PIP3BP. Generation of PIP3 in the nucleus was detected in H2O2-treated 293T cells, nerve growth factor (NGF)-treated PC12 cells, and platelet-derived growth factor (PDGF)-treated NIH 3T3 cells. Translocation of phosphatidylinositol 3-kinase (PI 3-kinase) to the nucleus and enhanced activity of PI 3-kinase in the nucleus fraction were observed after H2O2 treatment of 293T cells, suggesting that PI 3-kinase can be activated in the nucleus as well as in the membrane after appropriate stimulation of the cells. Co-expression of the constitutively active PI 3-kinase with PIP3BP resulted in exportation of the protein from the nucleus to the cytoplasm, suggesting that PIP3BP can function as a PIP3-binding protein in the intact cells. These results imply that there may be an unknown function of PI 3-kinase in the nucleus.     

Microstructure control and high temperature properties of TiAl base alloys

An equiaxed fine grain structure, a γ grain structure with the precipitated ₂ laths, and a fully lamellar structure were obtained by the microstructure control using thermomechanical processing and heat treatment. The key to obtaining the equiaxed fine grain structure using isothermal forging is to decompose the lamellar structure and then produce the fine grain microstructure through dynamic recrystallization. TiAl base alloys consisting of fine equiaxed grains, in particular, Ti-39Al-9V consisting of the γ and B2 phases exhibited superplastic elongation of more than 600% at 1423 K. Creep rupture properties of TiAl binary alloys with various microstructures were studied in purified He in the temperature range from 1073 to 1373 K. Above 1173 K the precipitated ₂ phase improved the steady state creep rate and creep life. At 1023 K, the ₂ phase did not improve the creep rate, although the steady state creep rate decreased and the creep life increased as the γ grain size increased.     

Characterization of monofunctional catalase KatA from radioresistant bacterium Deinococcus radiodurans

Catalase plays a key role in protecting cells against toxic reactive oxygen species. Here we report on the cloning, purification and characterization of a catalase (KatA, DR1998) from the extremely radioresistant bacterium Deinococcus radiodurans. The size of purified D. radiodurans KatA monomer was 65 kDa while gel filtration revealed that the size of the enzyme was 240 kDa, suggesting that KatA formed a homotetramer in solution. Purified KatA displayed a final specific activity of 68,800 U/mg of protein. The catalase activity of KatA was inhibited by sodium azide, sodium cyanide and 3-amino-1,2,4-triazole. The absorption spectrum of KatA exhibited a Soret band at 408 nm. The position of the spectral peak remained unchanged following reduction of KatA with dithionite. No peroxidase activity was found for KatA. These results demonstrate that D. radiodurans KatA is a typical monofunctional heme-containing catalase. The stability of KatA with respect to H2O2 stress was superior to that of commercially available Aspergillus niger and bovine liver catalases. The relative abundance of KatA in cells in addition to the H2O2 resistance property may play a role in the survival strategy of D. radiodurans against oxidative damage.