Anti-cell death activity promotes pulmonary metastasis of melanoma cells

Bcl-2 inhibits apoptosis from a variety of stimuli, and a Bcl-2-binding protein BAG-1 also functions in protection from apoptosis in concert with Bcl-2. Here, we provide evidence that prolonged cell survival introduced by overexpression of Bcl-2 or BAG-1 proteins strongly promotes experimental pulmonary metastasis of melanoma B16-BL6 cells. In murine melanoma cell line B16-BL6, gene transfer-mediated expression of the Bcl-2 or BAG-1 led to prolonged cell survival against serum-starved apoptosis in vitro. The Bcl-2-expressing B16 cells, B16-Bcl-2 and the BAG-1-expressing B16 cells, B16-BAG-1 strongly enhanced pulmonary metastasis in allogenic BALB/c nude mice and whole lung weights were increased by 2.4-fold and 1.4-fold, respectively, compared with control transfectants, suggesting that Bcl-2 is a stronger positive modulator of metastasis. When the viable B16-Bcl-2 and control transfectants were injected subcutaneously into BALB/c nude mice, the colony numbers of pulmonary metastasis of the B16-Bcl-2 transfectant increased by 5.6-fold compared with the control transfectants. These enhanced metastatic potentials in the B16-Bcl-2 and the B16-BAG-1 transfectants were well correlated with anti-cell death activity against serum-starvation and enhanced cell viability on limiting dilution. Analysis of the transfectants however revealed that their growth rates, invasive ability and cell motility were not significantly altered by overexpression of either Bcl-2 or BAG-1 proteins. Taken together, these studies demonstrate that prolonged cell survival is a crucial factor to promote metastasis of melanoma, thereby contributing to tumor progression.     

Characterization of bacteriocin N15 produced by Enterococcus faecium N15 and cloning of the related genes

Enterococcus faecium N15 was isolated from nuka (Japanese rice-bran paste), which is utilized as starter in the fermenting of vegetables, and was found to produce a bacteriocin that exhibited a broad spectrum of activity, including activity against Listeria monocytogenes and Bacillus circulans JCM2504. The bacteriocin was sensitive to proteases (alpha-chymotrypsin, proteinase K, trypsin, and pepsin) and alpha-amylase, but it was resistant to lipase. The bacteriocin was resistant to heat treatment at 100 degrees C for 2 h, but its activity was completely lost after autoclaving at 121 degrees C for 15 min. It was active over a wide pH range from 2.0 to 10.0. The bacteriocin showed bactericidal activity against Lactobacillus sake JCM1157 at a concentration of 40 AU/ml. Its molecular weight was estimated by SDS-PAGE to be about 3-5 kDa. PCR primers were designed based on the conserved amino acid sequences of class IIa bacteriocins. A 3-kb DNA fragment was amplified and three open reading frames (ORFs) were found. The first encodes a probable immunity protein of 103 amino acid residues and shows complete homology with the putative immunity protein of E. faecium DPC1146. The second and third ORFs respectively encode a probable transposase gene and an inducing factor. The upstream region of the immunity gene, in which the bacteriocin structural gene is located, was amplified. A homology search revealed that the bacteriocin produced by E. faecium N15 exhibits complete identity to enterocin A, a bacteriocin produced by E. faecium DPC1146. PCR using the primers designed in this study is a rapid and sufficient method of screening for bacteriocin-producing strains.     

Simulation of the effect of magnification and image-rotation variations on three-dimensional reconstruction

Tilting a specimen may change the magnification and the image rotation for electron-microscopic images, because re-focusing with an objective lens is required to correct for the height variation of the tilted specimen. Computer simulation is performed to analyse the effect of variations in the magnification and the image rotation on the three-dimensional reconstruction in electron tomography.     

Partial removal of PCDD/Fs, coplanar PCBS, and PCBS from municipal solid waste incineration fly ash by a column flotation process

Municipal solid waste incineration (MSWI) fly ash has recently attracted much attention because of its large quantity and enrichment of high toxic combustion generating organohalogen contaminants such as polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (coplanar PCBs), and polychlorinated biphenyls (PCBs). Since the organohalogen contaminants in MSWI fly ash are known to be enriched in the unburnt carbon (UC) fraction, the organohalogen contaminants can therefore be removed by the removal of UC. In this research, we used a modified column flotation technique to remove the organic contaminants from MSWI fly ash. UC was removed for 27.7% under the flotation condition without chemical flotation aids. The removal efficiencies of UC, PCDD/Fs, coplanar PCBs, and PCBs are further improved by adding flotation aids during the flotation process. UC was removed for 49.0% by adding a collector assistant with a HLB value of 13.5 and a concentration in the kerosene of 3% during the flotation process. In addition,the UC removal efficiencies are increased with the decrease of the diameter of the micropores in the gas spargers. By optimizing the flotation condition, 41.9% total PCDD/Fs, 40.8% coplanar PCBs, and 44.1% PCBs with 64.0% UC have been successfully removed from MSWI fly ash. The total toxic equivalent (TEQ) of the fly ash was decreased from 6.2 ng/g to 4.2 ng/g in the residue.     

Thermo-optic dispersion formula for AgGaS2

AgGaS(2) has been found to be 90 degrees phase matchable at 192 degrees C for sum-frequency mixing between the output of the KTP parametric oscillator at 3.2627 mum and its pump source at 1.0642 mum. The thermo-optic dispersion formula that can be used for good reproduction of the temperature-dependent phase-matching conditions thus far reported in the literature as well as our new data for second-harmonic generation of a CO(2) laser and its harmonics at 1.7652-5.2955 mum is presented.     

Measurement of electron transmission through tilted thick specimens with an ultrahigh voltage electron microscope

Electron transmission through tilted epoxy-resin film specimens with thicknesses of 1 and 5 microm has been measured under different accelerating voltages and diameters of the objective aperture in a 3 MV ultrahigh voltage electron microscope. Measurements show that the linearity between the logarithmic electron transmission and effective thickness of the tilted specimens holds valid only for some smaller effective thicknesses. However, the higher accelerating voltage leads to a wider linear attenuation range. The linear range can reach 7 microm for the 3 MeV incident electrons. Moreover, the measurements were analyzed with the electron-scattering theory. Actually, the high-energy electrons are capable of penetrating much thicker specimens, but many electrons elastically scattered at large angles cannot be collected as useful signals because of being blocked by the objective aperture in the mode of amplitude contrast. The need for further investigations was suggested to determine theoretically the electron transmission through a thick low-Z specimen and its exponential attenuation range.     

An automatic method of detecting and tracking fiducial markers for alignment in electron tomography

We presented an automatic method for detecting and tracking colloidal gold fiducial markers for alignment in electron tomography (ET). The second-order derivative of direction was used to detect a fiducial marker accurately. The detection was optimized to be selective to the size of fiducial markers. A preliminary tracking result from the normalized correlation coefficient was refined using the detector. A constraint model considering the relationship among the fiducial markers on different images was developed for removing outlier. The three-dimensional positions of the detected fiducial markers and the projection parameters of tilt images were calculated for post process. The accuracy of detection and tracking results was evaluated from the residues by the software IMOD. Application on transmission electron microscopic images also indicated that the presented method could provide a useful approach to automatic alignment in ET.     

日本污泥处置与资源化利用趋势

为了给我国污泥处理与处置及资源化技术提供更多的经验参考,对日本的污泥处置与资源化情况进行了文献调研和实地考察,重点分析了下水道污泥的各种处理与处置技术的发展情况、资源化及其发展趋势。日本因其特殊的国情和气候条件,目前的污泥资源化技术还是以污泥焚烧后产生的飞灰制作建筑材料为主,这可以作为我国很多大型城市污泥处理与处置技术的借鉴。