Development of Laser Ionization Techniques for Evaluation of the Effect of Cancer Drugs Using Imaging Mass Spectrometry

Recently, combined therapy using chemotherapy and photodynamic therapy (PDT) has been proposed as a means of improving treatment outcomes. In order to evaluate the efficacy of combined therapy, it is necessary to determine the distribution of the anticancer drug and the photosensitizer. We investigated the use of imaging mass spectrometry (IMS) to simultaneously observe the distributions of an anticancer drug and photosensitizer administered to cancer cells. In particular, we sought to increase the sensitivity of detection of the anticancer drug docetaxel and the photosensitizer protoporphyrin IX (PpIX) by optimizing the ionization-assisting reagents. When we used a matrix consisting of equal weights of a zeolite (NaY5.6) and a conventional organic matrix (6-aza-2-thiothymine) in matrix-assisted laser desorption/ionization, the signal intensity of the sodium-adducted ion of docetaxel (administered at 100 μM) increased about 13-fold. Moreover, we detected docetaxel with the zeolite matrix using the droplet method, and detected PpIX by fluorescence and IMS with α-cyano-4-hydroxycinnamic acid (CHCA) using the spray method.     

Determination of histamine in fish and fish products by tandem solid-phase extraction

A simple and practical method was developed for the determination of histamine in fish and fish products by solid-phase extraction and fluorescence derivatization. Histamine was extracted with trichloroacetic acid. The extract was neutralized and diluted with phosphate buffer (pH 6.8), and cleaned up with a tandem-connected octadecyl silica (ODS) and strong cation exchange silica (SCX) cartridge. After removal of the solvent, histamine was derivatized with fluorescamine and analyzed by ion-paired reversed-phase high-performance liquid chromatography with fluorescence detection. Recovery tests of histamine from six kinds of fish and fish products showed acceptable recovery (83-92%) with low relative standard deviation (less than 5%). This method could be useful for determination of histamine in fish.     

Mechanism of elongation of gold or silver nanoparticles in silica by irradiation with swift heavy ions

It has been reported that elongated Au nanoparticles oriented parallel to one another can be synthesized in SiO₂ by ion irradiation. Our aim was to elucidate the mechanism of this elongation. We prepared Au and Ag nanoparticles with a diameter of 20 nm in an SiO₂ matrix. It was found that Au nanoparticles showed greater elongated with a higher flux of ion beam and with thicker SiO₂ films. In contrast, Ag nanoparticles split into two or more shorter nanorods aligned end to end in the direction parallel to the ion beam. These experimental results are discussed in the framework of a thermal spike model of Au and Ag nanorods embedded in SiO₂. The lattice temperature exceeds the melting temperatures of SiO₂, Au and Ag for 100 ns after one 110 MeV Br¹⁰⁺ ion has passed through the middle of an Au or Ag nanorod.     

Lightning occurrence data observed with lightning location systems of electric power companies in Japan: 2009–2013

Nine electric power companies in Japan have been collecting lightning data with lightning location systems. Five years of the lightning data (2009–2013) are summarized and analyzed in this paper. The recent annual number of lightning flashes of which the current is more than 10 kA in Japan is around 1 million. The variations of lightning occurrence characteristics by areas, seasons, and so on, are clarified. Meteorological conditions that may affect lightning occurrence are also discussed. Furthermore, we show that there is a clear correlation between the number of lightning strikes and the outage rate of transmission lines. © 2016 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Time dependent changes occurring in rat liver microsomes upon lipid peroxidation

Steady-state fluorescence anisotropy of diphenylhexatriene and n-(9-anthroyloxy)stearic acids (n=2,12) in rat liver microsomes showed a marked increase in the early stages of enzymatically or non-enzymatically induced lipid peroxidation. The changes in fluorescence anisotropy occurred in parallel with the formation of thiobarbituric acid-reactive substances (TBA-RS). Parallel to these changes, the fluorescence emitted from peroxidized microsomes increased markedly in the early stages of lipid peroxidation. In contrast to the changes in the fluorescence anisotropy and in the formation of TBA-RS, the fluorescence showed a continuing increase over the three hr period of lipid peroxidation. Glucose-6-phosphatase was inactivated in the early stages of lipid peroxidation, whereas NADH-cytochrome b₅ reductase underwent a slow deactivation over three hr. The apparently slow deactivation of the peripheral protein may be explained by the formation of fluorescent substances.     

Incorporation of Halogenated Amino Acids into Antibody Fragments at Multiple Specific Sites Enhances Antigen Binding

Expansion of the amino-acid repertoire with synthetic derivatives introduces novel structures and functionalities into proteins. In this study, we improved the antigen binding of antibodies by incorporating halogenated tyrosines at multiple selective sites. Tyrosines in the Fab fragment of an anti-EGF-receptor antibody 059–152 were systematically replaced with 3-bromo- and 3-chlorotyrosines, and simultaneous replacements at four specific sites were found to cause a tenfold increase in the affinity toward the antigen. Structure modeling suggested that this effect was due to enhanced shape complementarity between the antigen and antibody molecules. On the other hand, we showed that chlorination in the constant domain, far from the binding interface, of Rituximab Fab also increased the affinity significantly (up to 17-fold). Our results showed that antigen binding is tunable with the halogenation in and out of the binding motifs.