ABSTRACT
A high-energy bench-top energy dispersive X-ray diffraction (EDXRD) system for 3-dimensional mapping of the crystalline structure and phase transformations in steel is described, for which preliminary data and system development are presented here. The use of precision tungsten slit screens with up to 225 keV X-rays allows for diffraction through samples of 304 L austenitic stainless steel of thickness 3-10 mm, while sample positioning is carried out with a precision goniometer and translation stage system.
Subject(s)
Crystallography/methods , Steel/chemistry , Tomography, X-Ray/instrumentation , Tomography, X-Ray/methods , Equipment Design , Equipment Failure Analysis , Reproducibility of Results , Sensitivity and Specificity , X-RaysABSTRACT
A trypsin inhibitor, with an N-terminal sequence highly homologous to those of 8-kDa Bowman-Birk trypsin inhibitors, was isolated from the seeds of Hokkaido large black soybeans. The trypsin inhibitor was unadsorbed on SP-Sepharose but adsorbed on DEAE-cellulose and Mono Q. It inhibited proliferation in breast cancer (MCF-7) cells and hepatoma (Hep G2) cells with an IC50 of 35 and 140 microM, respectively. The trypsin inhibitory activity of the inhibitor was completely preserved after exposure to temperatures up to 100 degrees C for 30 min and to the pH range 2-13 for the same duration. The trypsin inhibitor inhibited HIV-1 reverse transcriptase with an IC50 of 38 microM, but was devoid of antifungal activity toward Fusarium oxysporum and Mycosphaerella arachidicola.
Subject(s)
Cell Proliferation/drug effects , Glycine max/chemistry , Trypsin Inhibitor, Bowman-Birk Soybean/isolation & purification , Antifungal Agents/pharmacology , Breast Neoplasms/pathology , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Humans , Inhibitory Concentration 50 , Japan , Seeds/chemistry , Trypsin Inhibitor, Bowman-Birk Soybean/pharmacology , Trypsin InhibitorsABSTRACT
Two 30-kDa proteins with N-terminal sequence homology to chitinases have been isolated from fruits of the emperor banana by using a protocol that involved (NH(4))(2)SO(4) precipitation, affinity chromatography on Affi-gel blue gel, ion exchange chromatography by fast protein liquid chromatography (FPLC) on Mono S and gel filtration by FPLC on Superdex 75. The proteins were adsorbed on Affi-gel blue gel and Mono S. They both inhibited mycelial growth in Fusarium oxysporum but not in Mycosphaerella arachidicola. The chitinase-like protein more strongly bound on Mono S was obtained with a slightly lower yield and exhibited a higher antifungal potency toward F. oxysporum when compared with the less strongly bound chitinase-like protein.
Subject(s)
Antifungal Agents/isolation & purification , Chitinases/isolation & purification , Musa/enzymology , Amino Acid Sequence , Antifungal Agents/pharmacology , Chitinases/pharmacology , Chromatography, Gel , Electrophoresis, Polyacrylamide Gel , Fusarium/drug effects , Inhibitory Concentration 50ABSTRACT
A 20-kDa protein with substantial N-terminal sequence homology to thaumatin-like proteins was isolated from ripe fruits of the emperor banana, Musa basjoo cv. 'emperor banana'. The isolation procedure entailed (NH(4))(2)SO(4) precipitation, ion exchange chromatography on DEAE-cellulose, and affinity chromatography on Affi-gel blue gel. The thaumatin-like protein inhibited mycelial growth in Fusarium oxysporum and Mycosphaerella arachidicola. However, it did not affect the mitogenic response of murine splenocytes or [methyl-(3)H] thymidine incorporation by tumor cells. The activity of HIV-1 reverse transcriptase was slightly inhibited.
Subject(s)
Antifungal Agents/pharmacology , Musa/chemistry , Plant Proteins/pharmacology , Amino Acid Sequence , Ammonium Sulfate/chemistry , Animals , Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Ascomycota/drug effects , Cells, Cultured , Chromatography, Affinity , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Fusarium/drug effects , HIV Reverse Transcriptase/antagonists & inhibitors , HIV Reverse Transcriptase/metabolism , Inhibitory Concentration 50 , Mice , Mice, Inbred C57BL , Microbial Sensitivity Tests , Molecular Sequence Data , Plant Proteins/chemistry , Plant Proteins/isolation & purification , Sequence Alignment , Spleen/cytology , Spleen/drug effectsABSTRACT
An apparatus and a procedure are described for the preconcentration of nanogram amounts of trace elements. Co-precipitation and pressure filtration confine the precipitate to a 1.27 mm diameter spot on a filter membrane. Trace elements in the 10-100 ng range are collected reproducibly and detected with high sensitivity by X-ray fluorescence, by use of a milliprobe instrument. Advantages of the technique for quantitative applications are discussed.
