On-machine measurement of a cylindrical surface with sinusoidal micro-structures by an optical slope sensor

This paper describes the measurement of a cylindrical surface with sinusoidal micro-structures over a large area on a diamond turning machine. The sinusoidal micro-structures, which are fabricated on the periphery surface of a cylinder by the fast tool servo-based diamond turning, are superposition of periodic sine-waves along the cylinder axis and the cylinder circumference with amplitudes of 100 nm and wavelengths of 100 μm, respectively. An optical two-dimensional (2D) slope sensor with a multi-spot light beam is developed for measurement of the 2D local slopes of the sinusoidal micro-structured surface. A cylindrical lens is employed in the sensor for removing the influence of the curvature of the cylinder surface. Experiments of fabrication and measurement of the sinusoidal micro-structured surface on an ultra-precision diamond turning machine are carried out.     

Effect of matrix's type on the dynamic properties for short fiber-elastomer composite

The dynamic moduli, E′ and E″, and tan δ for PET–CR, PET–EPDM, and PET–UR composites with unidirectional short fibers were studied as a function of temperature by using a Rheovibron. The temperature dependence of tan δ showed three peaks for PET–elastomer composites. The peaks at the low temperature corresponded to the main dispersion of the respective matrixes and the peak at about 140°C to the α-dispersion of PET fiber. A small and broad peak observed at a temperature between 60 and 120°C may be caused by the relaxation of the interface region between fibers and matrix. The longitudinal storage modulus for the composite E was given by the parallel model as \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm E'}_\parallel = V_f \cdot E'_f + V_m \cdot E'_m $\end{document}, where E and E are the storage moduli for fiber and matrix and V_f and V_m are the volume fraction of fiber and matrix, respectively. In the transverse direction of fibers, the composite modulus E was expressed by the logarithmic law of mixing as follows: \documentclass{article}\pagestyle{empty}\begin{document}$ \log E'_ \bot = V_f \cdot \log E'_f + V_m \cdot \log E'_m $\end{document}. The peak values of tan δ from the main dispersion of the respective matrixes were given by the equation, (tan δ_⊥max)_c/(tan δ_max)_m 1 ? β · V_f, where (tan δ_⊥max)_c and (tan δ_max)_m are the maximum values of the loss tangent for the composite and matrix, respectively, and β is coefficient depending on matrix's type. The β value of PET–CR composite is the largest one among those of the composites.     

Characterization of Aspergillus oryzae glycoside hydrolase family 43 β-xylosidase expressed in Escherichia coli

This is the first report of glycoside hydrolase family 43 β-xylosidase from Aspergillus oryzae. To characterize this enzyme, the recombinant enzyme was expressed in Escherichia coli. Unlike known β-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline pH.     

Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907

Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5'-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.     

Novel screening system for protein-protein interactions by bimolecular fluorescence complementation in Saccharomyces cerevisiae

For high-throughput screening of protein-protein interactions, we have developed a novel yeast screening system using Bimolecular fluorescence complementation (BiFC). Two yeast plasmids, in which genes of heterodimerized peptides LZA and LZB were each fused with those of non-fluorescent half fragments of Kusabira-Green mutant (mKG2), were transformed into a- and α-type yeast, respectively. Mating of them gave a library, which was screened by following green fluorescence resulted from LZA-LZB interaction. The method showed potential ability to detect the positive clones from a model library, in which green-fluorescent and non-fluorescent yeast was mixed in a ratio of 1:675.     

Determination of ethychlozate and its degradation product in fruits by HPLC and LC/MS

A method was developed for the analysis of ethychlozate (CIE) and its decomposition compound, 5-chloro-3(1H)-indazolylacetic acid (CIA) in fruits by HPLC and LC/MS. The sample was homogenized with 1 mol/L HC1, and CIE and CIA were extracted with 5 mol/L HCl and acetone. They were extracted from the acetone extract with diethylether-n-hexane (2:1). CIE was hydrolyzed to CIA with methanol-4 mol/L KOH (1:1). The solution was made acidic, and CIA was extracted with diethylether-n-hexane (2:1). The extract was cleaned up on a silica gel column. CIA was determined by HPLC-UV and LC/MS (Scan or SIR). Four fruits were spiked with CIE or CIA at 0.5 microgram/g and analyzed by the proposed method with HPLC. The average recoveries were 77.2-83.2% for CIE and 71.2-89.2% for CIA. The concentrations determined by LC/MS were 10-25% higher than the values by HPLC. The limit of detection (LOD) of CIA standard solution by HPLC corresponds to 0.015 microgram/g of CIE in the sample. In the same way, the LOD of CIA by LC/MS (SIR) corresponds to 0.009 microgram/g of CIE in the sample.     

Photodegradation of tetrahalobisphenol-A (X = Cl, Br) flame retardants and delineation of factors affecting the process

The photoassisted degradation (HPLC-UV absorption), dehalogenation (HPLC-IC) and mineralization (TOC decay) of the flame retardants tetrabromobisphenol-A (TBBPA) and tetrachlorobisphenol-A (TCBPA) were examined in UV-irradiated alkaline aqueous TiO₂ dispersions (pH 12), and for comparison the parent bisphenol-A (BPA, an endocrine disruptor) in pH 4–12 aqueous media to assess which factor impact most on the photodegradative process. Complete degradation (2.7–2.8 × 10⁻² min⁻¹) and dehalogenation (1.8 × 10⁻² min⁻¹) of TBBPA and TCBPA occurred within 2 h of UV irradiation, whereas only 45–60% mineralization (2.3–2.7 × 10⁻³ min⁻¹) was complete within 5 h for the flame retardants at pH 12 and ca. 80% for the parent BPA. Factors examined in the pH range 4–12 that impact the degradation of BPA were the point of zero charge of TiO₂ particles (pH_pzc; electrophoretic method), particle or aggregate sizes of TiO₂ (light scattering), and the relative number of OH radicals (as DMPO–OH adducts; ESR spectroscopy) produced in the UV-irradiated dispersion. Dynamics of BPA degradation (2.0–2.4 × 10⁻² min⁻¹) were pH-independent and independent of particle/aggregate size, but did correlate with the number of OH radicals, at least at pHs 4 to 8–9, after which the rates decreased somewhat at pH > 9 with decreasing adsorption owing to Coulombic repulsive forces between the very negative TiO₂ surface and the anionic forms of BPA (pK_as ca. 9.6–11.3), even though the number of OH radicals continued to increase at the higher pHs.     

Novel synthesis of poly(methyl methacrylate) brush encapsulated silica particles

Silica (SiO₂)‐crosslinked polystyrene (PS) particles possessing photofunctional N,N‐diethyldithiocarbamate (DC) groups on their surface were prepared by the free‐radical emulsion copolymerization of a mixture of SiO₂ (diameter = 20 nm), styrene, divinyl benzene, 4‐vinylbenzyl N,N‐diethyldithiocarbamate (VBDC), and 2‐hydroxyethyl methacrylate with a radical initiator under UV irradiation. In this copolymerization, the inimer VBDC had the formation of a hyperbranched structure by a living radical mechanism. The particle sizes of such core–shell structures [number‐average particle diameter (D_n) = 35–40 nm] were controlled by the variation of the feed amounts of the monomers and surfactant, or emulsion system. The size distributions were relatively narrow (weight‐average particle diameter/D_n ≈ 1.05). These particles had DC groups on their surface. Subsequently, poly(methyl methacrylate) brush encapsulated SiO₂ particles were synthesized by the grafting from a photoinduced atom transfer radical polymerization approach of methyl methacrylate initiated by SiO₂‐crosslinked PS particles as a macroinitiator. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Formation mechanism for hexagonal-structured self-assemblies of nanocrystalline titania templated by cetyltrimethylammonium bromide

Hexagonal-structured self-assemblies of nanocrystalline (anatase) titania templated by cetyltrimethylammonium bromide (C(16)H(33)N(CH(3))(3)Br; CTAB) (Hex-ncTiO(2)/CTAB Nanoskeleton) were formed after mixing of aqueous solutions containing CTAB spherical micelles and titanium oxysulfate acid hydrate (TiOSO(4).xH(2)SO(4).xH(2)O) as a titania precursor in the absence of any other additives. Formation mechanism of the Hex-ncTiO(2)/CTAB Nanoskeleton was examined in terms of the reaction temperature, titania precursor/CTAB mixing ratio, surfactant type, electrostatic interaction, micelle formation and molecular component. We found that crystal growth of crystalline (anatase) titania (polymorphic crystallization) was promoted with higher temperature and lower titania precursor content in aqueous solutions. In addition, we revealed that the crystalline (anatase) titania was formed in polycation, poly(allylamine hydrochloride ([CH(2)CH(CH(2)NH(2))HCl](n); PAH), and formamide (HCONH(2)) solutions. On the other hand, no titania formation was observed in anionic systems such as sodium dodecyl sulfate (CH(3)(CH(2))(11)OSO(3)Na; SDS) and poly(sodium 4-styrenesulfonate ([C(8)H(7)SO(3)Na](n); PSSS). This indicates that hydrolysis reaction of the titania precursor is initiated by not only cations but also nitrogen atoms in molecules and polymers. Hexagonally structure was formed in only cationic surfactant micellar solutions but not in polycation solutions and formamide.     

Synthesis and application of fluorescein- and biotin-labeled molecular probes for the chemokine receptor CXCR4

The design, synthesis, and bioevaluation of fluorescence- and biotin-labeled CXCR4 antagonists are described. The modification of D-Lys8 at an epsilon-amino group in the peptide antagonist Ac-TZ14011 derived from polyphemusin II had no significant influence on the potent binding of the peptide to the CXCR4 receptor. The application of the labeled peptides in flow cytometry and confocal microscopy studies demonstrated the selectivity of their binding to the CXCR4 receptor, but not to CXCR7, which was recently reported to be another receptor for stromal cell-derived factor 1 (SDF-1)/CXCL12.