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.     

Particle velocity and solids circulation rate in a jet-spouted bed

Radial distribution of particle velocities, solids circulation rate and radial variation in bed voidage in a jet-spouted bed were investigated in a 0.2 m diameter conical-cylindrical column. The upward particle velocity is strongly dependent on gas velocity and its radial profile is the Gaussian. The solids circulation rates are of the same order of magnitude as those in a conventional spouted bed and are proportional to the gas velocity. Estimation of the variation in bed voidage in the radial direction revealed that the annulus voidage of about 0.90 was greater than the spout voidage of around 0.70. This tendency is contrary to that in a conventional spouted bed.     

Morphological study by TEM on uniaxially oriented thin films of PET, PEN and their blends

Uniaxially oriented thin films of poly(ethylene terephthalate) (PET), poly(ethylene 2,6-naphthalene dicarboxylate) (PEN) and their blends were prepared by applying shear strain to their respective melts, and the resulting morphologies were studied by transmission electron microscopy. Selected-area electron diffraction of each film revealed well-defined uniaxial orientation of polymer chains in the shearing direction. In the uniaxially oriented thin film of PEN, stacked-lamellar structure with the average long period of 27 nm consisting of a crystalline region about 15 nm thick and an amorphous one about 12 nm thick was found here and there in the dark-field image: PEN-type. On the other hand, stacked-lamellar structure was rarely observed in the case of PET: PET-type. In PET/PEN blends, the morphologies changed from the PET-type to the PEN-type with increasing content of PEN.     

Comparative study on the preparation and properties of radiation‐grafted polymer electrolyte membranes based on fluoropolymer films

In this study the fluoropolymers, poly(ethylene‐co‐tetrafluoroethylene) (ETFE) and poly(vinylidene fluoride) (PVDF) films, together with the radiation‐induced crosslinked polytetrafluoroethylene (cPTFE) film were compared on the basis of their preparation and properties of radiation‐grafted polymer electrolyte membranes. The polymer electrolyte membranes were prepared by radiation grafting of styrene into the base films and subsequent sulfonation. The proton conductivity and chemical stability of the three types of membranes with a similar ion exchange capacity (IEC) near 1.0 mmol/g were investigated and are discussed in detail. Although the ETFE‐based polymer electrolyte membrane was relatively more stable, its proton conductivity was lower than those of the PVDF‐ and cPTFE‐based membranes. On the other hand, the cPTFE‐based membrane showed a significantly higher proton conductivity, but its chemical stability was shorter than that of the ETFE‐based membrane. It is considered that the difference in the preparation and properties of the polymer electrolyte membranes was due to the difference in the degree of crystallinity as well as in the chemical structure of the fluoropolymer base films. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 1966–1972, 2007     

Effect of crosslinkers on the preparation and properties of ETFE‐based radiation‐grafted polymer electrolyte membranes

This study concerns a comparative study of three crosslinkers, divinylbenzene (DVB), 1,2‐bis(p,p‐vinylphenyl)ethane (BVPE), and triallyl cyanurate (TAC) crosslinked poly(ethylene‐co‐tetrafluoroethylene) (ETFE)‐based radiation‐grafted membranes, which were prepared by radiation grafting of p‐methylstyrene onto ETFE films and subsequent sulfonation. The effect of the different types and contents of the crosslinkers on the grafting and sulfonation, and the properties such as water uptake, proton conductivity, and thermal/chemical stability of the resulting polymer electrolyte membranes were investigated in detail. Introducing crosslink structure into the radiation‐grafted membranes leads to a decrease in proton conductivity due to the decrease in water uptake. The thermal stability of the crosslinked radiation‐grafted membranes is also somewhat lower than that of the noncrosslinked one. However, the crosslinked radiation‐grafted membranes show significantly higher chemical stability characterized in the 3% H₂O₂ at 50°C. Among the three crosslinkers, the DVB shows a most pronounced efficiency on the crosslinking of the radiation‐grafted membranes, while the TAC has no significant influence; the BVPE is a mild and effective crosslinker, showing the moderate influence between the DVB and TAC crosslinkers. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 4565–4574, 2006     

Tribological Characteristics of SiC Ceramics in High-Temperature and High-Pressure Water

The effects of temperature and sliding speed on the tribological behavior of a SiC ceramic by sliding on the same material in deoxygenated water were investigated from room temperature to 300°C under the corresponding saturated vapor pressures. The friction coefficient and specific wear rates of both plates and disks increased at elevated temperatures at all sliding speeds, but decreased with increasing sliding speed at 120° and 300°C. Fine mirrorlike worn surfaces were observed without wear debris under all sliding conditions. The wear mechanism appears to consist of hydrothermal oxidation of SiC and dissolution of reaction products such as silica.     

Hot isostatic pressing of tetragonal ZrO2 solid-solution powders prepared from acetylacetonates in the system ZrO2-Y2O3-Al2O3

In compositions having ZrO₂/Y₂O₃=(74.25–71.25)/(0.75–3.75) (mol% ratio) with 25 mol% Al₂O₃, metastable t-ZrO₂ solid solutions crystallize at 780° to 860°C from amorphous materials prepared by the simultaneous hydrolysis of zirconium, yttrium and aluminium acetylacetonates. Hot isostatic pressing has been performed for 1 h at 1130 and 1230°C under 196 MPa using their powders. Two kinds of material are fabricated: (i) perfect ZrO₂ solid-solution ceramics and (ii) composites of ZrO₂ solid solution and -Al₂O₃. Their mechanical properties are examined, in connection with microstructures and t/m ZrO₂ ratios. Composites with a homogeneous dispersed -Al₂O₃ derived from solid-solution ceramics result in a remarkable increase of strength.     

Methanation of CO2 with the oxygen-deficient Ni(II)-ferrite under dynamic conditions

The continuous methanation of CO₂ has been accomplished over hydrogen-reduced Ni(II)-bearing ferrite (Ni_xFe_3–xO_4–; x=0.39, > 0) in a mixed gas flow of CO₂ and H₂ at 250–375 °C. The yield and the selectivity for the methanation were larger than 50% and 95%, respectively, at the initial stage of the process. They decreased to 31% and 89%, respectively, after 6 h methanation. The innovative results can be ascribed to the use of the new material; hydrogen-reduced Ni(II)-bearing ferrite. Its formation was evinced by chemical analyses and the increase in the lattice constant; the lattice constant of the Ni(II)-bearing ferrite (a₀ 0.8375 nm) was enlarged to 0.8379 nm by hydrogen reduction. The enlarged lattice constant was not changed during the methanation. These findings suggest that the methanation occurs at the oxygen-deficient site of the hydrogen-reduced Ni(II)-bearing ferrite, as well as the formation of water by combination of the incorporated oxygens with hydrogen. The methanation consists of three steps of the elementary reaction. First, the oxygen-deficient sites are formed by hydrogen reduction; second, CO₂ is reduced to elementary carbon and two oxygen ions which are incorporated into the oxygen-deficient sites; and third, the carbon deposited on the surface of the reduced ferrite is selectively hydrogenated to CH₄.     

The effects of amino-acid mutations on specific interactions between urokinase-type plasminogen activator and its receptor: Ab initio molecular orbital calculations

During cancer invasion, the binding of urokinase-type plasminogen activator (uPA) to its receptor (uPAR) on the surface of a cancer cell is considered a trigger for invasion. Here, we present a stable structure of the solvated complex formed between uPA and uPAR (uPA-uPAR) and investigate the specific interactions between uPA and uPAR by ab initio fragment molecular orbital (FMO) calculations. The result indicates that the electrostatic interactions between the charged amino acid residues existing in both uPA and uPAR make a large contribution to the binding between uPA and uPAR. In particular, Lys23, Lys46, Lys98 and Lys61 of uPA are found to have strong attractive interactions with uPAR. To elucidate the effect of these residues on the interactions between uPA and uPAR, we substituted each of them with the uncharged amino acid Leu and investigated the interactions between the mutated uPA and wild-type uPAR. The interaction energies indicate that Lys46 and Lys98, which bind uPA to the rim of the central ligand-binding cavity of uPAR, make greater contributions to the binding between uPA and uPAR than Lys23, which is positioned at the bottom of the ligand-binding cavity of uPAR. The effect of hydrating water molecules located between uPA and uPAR is also investigated to be significant for the specific interactions between uPA and uPAR. These results are expected to be informative for developing new peptide antagonists that block the binding of uPA to uPAR.