Time-resolved DXAFS study on the reduction processes of Cu cations in ZSM-5

The time-resolved reduction process of copper cations in/on ZSM-5 during temperature-programmed reduction (300–700 K) was studied by energy-dispersive X-ray absorption fine structure (DXAFS) as well as transmission electron microscopy (TEM). Two Cu-ZSM-5 samples with different Cu loadings were prepared by an ion-exchange method. The Cu K-edge DXAFS spectra for isolated Cu₂₊ species in the channels of ZSM-5 and CuO particles on the outer surfaces of ZSM-5 were recorded at an interval of 1 s during the reduction. The curve fitting analysis of the EXAFS data and the XANES analysis revealed that the isolated Cu₂₊ species in the channels were reduced stepwise. They were reduced to isolated Cu₊ species at 400–450 K and the Cu₊ species to Cu₀ metallic clusters at 550–650 K. Small clusters like Cu₄ were initially formed, followed by particle growth. A small part of them went out to the outer surfaces of ZSM-5 during the reduction. In contrast, CuO particles on the outer surfaces were reduced directly to Cu₀ metallic particles around 450 K.     

A Novel Artificially Humanized Anti-Cripto-1 Antibody Suppressing Cancer Cell Growth

Cripto-1 is a member of the EGF-CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti-Cripto-1 artificial antibody and assessed the recognition to the antigen and the potential to suppress the growth of cancer stem cells. First, single chain antibody clones were isolated by bio-panning with the affinity to recombinant Cripto-1 protein from our original phage-display library. Then, the variable regions of heavy chain VH and light chain VL in each clone were fused to constant regions of heavy chain CH and light chain CL regions respectively. These fused genes were expressed in ExpiCHO-S cells to produce artificial humanized antibodies against Cripto-1. After evaluation of the expression levels, one clone was selected and the anti-Cripto-1 antibody was produced and purified. The purified antibody showed affinity to recombinant Cripto-1 at 1.1 pmol and immunoreactivity to cancer tissues and cell lines. The antibody was available to detect the immunoreactivity in tissue microarrays of malignant tumors as well as in Cripto-1 overexpressing cells. Simultaneously, the antibody exhibited the potential to suppress the growth of human colon cancer derived GEO cells overexpressing Cripto-1 with IC₅₀ at approximately 110 nM. The artificially humanized antibody is proposed to be a good candidate to target cancer cells overexpressing Cripto-1.     

Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis

High-resolution melting (HRM) analysis has been improved and applied for the first time to quantitative analysis of enzymatic reactions. By using the relative ratios of peak intensities of substrates and products, the quantitativity of conventional HRM analysis has been improved to allow detailed kinetic analysis. As an example, the ligation of sticky ends through the action of T4 DNA ligase has been kinetically analyzed, with comprehensive data on substrate specificity and other properties having been obtained. For the first time, the kinetic parameters (k_obs and apparent K_m) of sticky-end ligation were obtained for both fully matched and mismatched sticky ends. The effect of ATP concentration on sticky-end ligation was also investigated. The improved HRM method should also be applicable to versatile DNA-transforming enzymes, because the only requirement is that the products have T_m values different enough from the substrates.     

A novel sphingophosphonolipid head group 1-hydroxy-2-aminoethyl phosphonate in Bdellovibrio stolpii

Members of the bacterial genus Bdellovibrio include strains that are free-living whereas others are known to invade and parasitize larger Gram-negative bacteria. The bacterium can synthesize several sphingophospholipid compounds including those with phosphoryl bonds as well as phosphonyl bonds. In the present study, the dominant sphingophosphonolipid component was isolated by column chromatography, and the long-chain bases, fatty acids, and polar head groups were identified by thin-layer and gas-liquid chromatographic procedures. The definitive structural identity of the sphingolipid was established by nuclear magnetic resonance and mass spectrometry of hydrolysis products and the intact compound. The compound was identified as N-2′-hydroxypentadecanoyl-2-amino-3,4-dihydroxyheptadecan-1-phosphono-(1-hydroxy-2-aminoethane).     

Influence of Cu metal on the domain structure and carrier mobility in single-layer graphene

We demonstrate that domain structure of single-layer graphene grown by ambient pressure chemical vapor deposition is strongly dependent on the crystallinity of the Cu catalyst. Low energy electron microscopy analysis reveals that graphene grown using a Cu foil gives small and mis-oriented graphene domains with a number of domain boundaries. On the other hand, no apparent domain boundaries are observed in graphene grown over a heteroepitaxial Cu(111) film deposited on sapphire due to unified orientation of graphene hexagons. The difference in the domain structures is correlated with the difference in the crystal plane and grain structure of the Cu metal. The graphene film grown on the heteroepitaxial Cu film exhibits much higher carrier mobility than that grown on the Cu foil.     

High quality epitaxial ZnO films grown on solid-phase crystallized buffer layers

High quality epitaxial ZnO films on sapphire (110) plane have been fabricated on ZnO homo-buffer layers crystallized via solid-phase epitaxially (SPE). The SPE-ZnO films are fabricated by annealing of amorphous ZnON (a-ZnON) films deposited by RF magnetron sputtering. During annealing, the a-ZnON films are oxidized and converted to ZnO crystal. X-ray diffraction (XRD) analysis shows that the resultant films are epitaxially grown on the sapphire substrates. By using the SPE-ZnO films as homo-buffer layers, the ZnO films with high crystallinity, which are deposited by RF magnetron sputtering, are fabricated. The full width at half-maximum of XRD patterns for 2θ-ω and ω scan of (002) plane are 0.094° and 0.12°, respectively, being significantly small compared with 0.24° and 0.55° for the films without buffer layers. Thus utilizing SPE buffer layers is very promising to obtain epitaxial ZnO films with high crystallinity.     

The accessory middle cerebral artery as a collateral blood supply

We describe two cases of acute embolic occlusion of the internal carotid artery and the middle cerebral artery in association with a patent accessory middle cerebral artery. Because of the presence of the accessory middle cerebral artery, the frontal lobe was salvaged to some extent, but it did not provide sufficient collateral blood supply to the middle cerebral artery territory to spare the rest of the frontal lobe, the temporal lobe, and the basal ganglia.     

Comparison of DNA copy numbers in original oral squamous cell carcinomas and corresponding cell lines by comparative genomic hybridization

We analyzed regional DNA copy numbers in 4 oral squamous cell carcinomas (SCCs) by using comparative genomic hybridization, and compared them with those in cell lines derived from the SCCs. In the original tumors, DNA copy number increases were observed on chromosomes 5p (4/4 cases), 8q (4/4), 20p (3/4), 3q (2/4), 5q (2/4), 7p (2/4), 7q (2/4), 11p (2/4), 11q (2/4) and 13q (2/ 4). Although most of these changes have been described previously for SCC tumors in the head and neck, the incidence of increases in 8q and 20p was much higher in the present study; this may be important in relation to cell line establishment, since 8q contains e-myc, which is involved in immortalization. No common chromosomal region with DNA copy number decreases was observed, except for 18q (2/4). When the original tumors and the cell lines were compared, their profiles were essentially similar with one exception. Further, there was no region that commonly changed in the cell lines, but not in the original tumors, suggesting that the DNA copy number changes observed in the cell lines mostly represent those of the original tumors.