Microscale analysis of glycosphingolipids by TLC blotting/secondary ion mass spectrometry: a novel blood group A-active glycosphingolipid and changes in glycosphingolipid expression in rat mammary tumour cells with different metastatic potentials

The glycosphingolipid compositions of rat mammary tumour cell lines with different metastatic potentials for the lung [a parental tumour cell line (MTC) and its subclones MTLn2 (a non metastatic subclone) and MTLn3 (a subclone with high metastatic potential to the lung)] were studied using a newly developed TLC blotting/secondary ion mass spectrometry system and crude glycosphingolipids obtained from 0.5-1 x 10(7) cells of each cell line. GM3 and GM2 were the major components of the MTC cell line, but they were very minor components in the MTLn2 and MTLn3 cell lines, GDla being the major ganglioside HexNAc-fucosyl-GMla was found in the MTLn2 cells by the TLC blotting/SIMS method, and the terminal sugar linkage was shown to be a blood group A-type structure by immunostaining. These findings suggest that the ganglioside is a novel type of blood group A-active ganglioside, GalNAc alpha 1-3(fuc alpha 1 -2)GMla. No blood group A-active lipid was present in MTLn3 cells, whereas Hex-GMla and neutral glycosphingolipids with more than 5 sugar residues were.     

Bis-(monoacylglyceryl) phosphate and acyl phosphatidylglycerol isolated from human livers of lipidosis induced by 4,4′-diethylaminoethoxyhexesterol

Bis-(monoacylglyceryl) phosphate and acyl phosphatidylglycerol were isolated from the liver of two patients with lipidosis induced by 4,4′-diethylaminoethoxyhexesterol. Identification was based upon the results of alkaline hydrolysis, acetolysis, IR spectrometry, and upon the determination of molar ratio of phosphorus-glycerol-ester. The contents of the bis-(monoacylglyceryl) phosphate were 10 and 16% total phospholipid phosphorus in them. The bis-(monoacylglyceryl) phosphate contained mainly docosahexaenoic (42%), oleic (29%), and linoleic acid (14%) and had the hemolytic activity of ca. one-eighth lysolecithin from egg yolk. Acidic lipids from the liver also were found to contain a lipid which is less polar than bis-(monoacylglyceryl) phosphate. The results of lipid analysis showed that the lipid possessed the structure of an acyl phosphatidylglycerol, and its content was ca. 2% total phospholipid phosphorus. Accumulation of 4,4′-diethylaminoethoxyhexesterol and its derivatives was found in clinical cases by thin layer chromatography and IR spectrometry. This fact suggested that human liver has an ability to metabolize the drug.     

Hydrothermal Formation of Hydroxyapatite Layers on the Surface of Type-A Zeolite

Type-A zeolite evenly covered with hydroxyapatite thin layers was prepared using hydrothermal treatment at 120°C for 8 h under autogenous pressure. The hydroxyapatite needlelike nanocrystals, 100–200 nm in diameter and 30 nm in thickness, were grown under the reaction between discharged Ca²⁺ ions from type-A zeolite and PO₄³⁻ ions in (NH₄)₃PO₄ solution. The preferential orientations of the c -axis of hydroxyapatite crystals perpendicular to a zeolite surface were observed using transmission electron microscopy. The crystal structure of type-A zeolite was not destroyed under the reaction, but the surface morphology was changed only with complete covering of scaly hydroxyapatite particles.     

Geo- and Hydro-Mechanical Evaluation of Slope Failure Induced by Torrential Rains in Northern-Kyushu Area,July 2009

Torrential rainfall in mid-July 2009 triggered numerous geodisasters such as slope failure and debris flow in Chugoku and Northern Kyushu areas of Japan. A number of slope failures and debris flows occurred in Yamaguchi and Fukuoka prefectures resulting in extensive damage to human life and infrastructure. One of the most serious geodisasters included a slope failure followed by debris flow at Sasaguri-machi and Fukuchi-machi, Fukuoka prefecture, Japan. This paper summarizes the results of geotechnical investigations on the geodisaster sites in Fukuoka prefecture. The geotechnical investigation included determining a series of grain size distributions, consistency limits and conducting direct box shear tests for collapsed soils collected at six disaster sites. The generation mechanisms of slope failure followed by debris flow were also investigated by analyzing the precipitation, topography, geology, and strength properties of the collapsed soils. Moreover, slope deformation and stability analyses were coupled with an unsaturated-saturated seepage analysis to investigate the slope failure mechanism. The main findings from the study are summarized as: The physical properties, such as the grain size distribution, the plastic limit and liquid limit of collapsed soils, are summarized and compared with the results of other failure slopes in the literature. The collapsed soil was characterized as being a well grained soil (the uniformity coefficient >50) and highly weathered (the ignition loss >5%), however, with regard to the liquid limit and plastic index, there were no remarkable findings. The original shear strength for collapsed soils with natural water content is relatively large and slope failure doesn't occur because the cohesion in the shear strength is induced by a suction force between the soil particles under unsaturated condition. However, water seepage into the soil induces a drastic decrease in the shear strength, which is mainly caused by a decrease in cohesion (losing suction) resulting from soil saturation. In addition, the drained/undrained condition in the shear process is also sensitive to shear strength. For example, both water seepage and the shear process with constant volume cause an approximate 30% reduction in shear strength for Fukuchi-machi and Sasaguri-machi soil samples. Therefore, the reduction of cohesive strength due to water seepage and the low permeability of the slope are the parameters which trigger geodisaster. Based on the results of slope deformation and a stability analyses which took the change in water pressure and cohesive strength into account, the geodisaster at Fukuchi-machi was simulated, it is reasonable to assume that the shallow failure near the top of slope occurred due to torrential precipitation of about 100 mm per hour which triggered a debris flow.     

The preparation of copper fine particle paste and its application as the inner electrode material of a multilayered ceramic capacitor

Well size-controlled copper fine particles (diameter: 100-300?nm) were used as the inner electrode material of multilayered ceramic capacitors (MLCCs). The particles were dispersed in terpineol to form a printing paste with 50?wt% copper particles. The MLCC precursor modules prepared by the layer-by-layer printing of copper and BaTiO(3) particles were cosintered. Detailed observation of the particles, paste, and MLCCs before and after sintering was carried out by electron microscopy. The sintering temperature of Cu-MLCC was as low as 960?°C. The permittivity of these MLCCs was successfully measured with the copper inner layers.     

Size-controlled oxidation-resistant copper fine particles covered by biopolymer nanoskin

Size-controlled oxidation-resistant copper fine particles were prepared from insoluble CuO micron-sized particles. The particle sizes were quite uniform and could be varied only by the concentration of the complex reagent from 45 to 175 nm. No template material was needed for size control. Gelatin was selected as the protective polymer. Addition of protease after formation of copper fine particles decomposed preferentially loop and tail parts of gelatin. The remaining nanoskin gelatin layer, covered on the particle surface, prevents oxidation of copper.     

Cloning and sequencing of the histidine decarboxylase gene from Photobacterium phosphoreum and its functional expression in Escherichia coli

The major causative agent of scombroid poisoning is histamine formed by bacterial decarboxylation of histidine. We reported previously that histamine was exclusively formed by the psychrotrophic halophilic bacteria Photobacterium phosphoreum in scombroid fish during storage at or below 10 degrees C. Moreover, histamine-forming ability was affected by two histidine decarboxylases (HDCs): constitutive and inducible enzymes. In this study, the gene encoding P. phosphoreum HDC was cloned into Escherichia coli and sequenced. A sequence analysis of the DNA corresponding to the hdc gene revealed an open reading frame of 1,140 bp coding for a pyridoxal-5'-phosphate-dependent HDC of 380 amino acid residues with a predicted molecular mass of 42.6 kDa. The HDC amino acid sequences formed a phylogenetic clade with strong bootstrap support and revealed high sequence similarities among the P. phosphoreum isolate and species of the family Enterobacteriaceae and a separate phylogenetic branch with the lowest sequence similarity between the isolate and the taxonomically closer Listonella anguillarum. The T7 promoter was used to overexpress the hdc gene in E. coli cells. The recombinant clone, E. coli BL21(DE3), displayed significant levels of HDC activity. The recombinant hdc gene was suggested to code the inducible HDC; therefore, the optimum reaction conditions of the recombinant HDC were similar to those of the inducible HDC in the P. phosphoreum isolate. In addition, a putative catabolite-repressor protein binding site, amino acid permease gene, and histidine-tRNA synthetase gene were found in flanking regions of the hdc gene.     

Behavior of Cu nanoparticles ink under reductive calcination for fabrication of Cu conductive film

Cu nanoparticle ink was prepared from Cu nanoparticles that were coated with a gelatin layer at an average diameter of 46 nm. The Cu nanoparticle ink was applied on the polyimide substrate. Conductive films were fabricated using the Cu nanoparticle ink with a two-step annealing process consisting of oxidative pre-heating at 200 °C under 10 ppm O₂-N₂ mixed gas flow and reductive calcination at 250 °C under 3 vol.% H₂-N₂ mixed gas flow showed a low resistivity of 5 μΩ cm. The hydrolysis of the remaining gelatin layer by H₂O vapor, which was formed during the reduction of the Cu oxide by 3 vol.% H₂-N₂ mixed gas, was suggested. The results suggest the possibility of the removal of the gelatin layer without oxidative pre-heating and simultaneous sintering of Cu nanoparticles in reductive calcination.