19 research outputs found

    The expression profile of M2 protein and AcGFP-M2 fusion protein.

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    <p>(<b>A</b>) Expression of AcGFP-M2 fusion protein in the presence or absence of amantadine. Experiment procedures are the same as described in a. Positions of M2 protein and AcGFP-M2 fusion protein are indicated by arrowheads. (B) Expression of M2 protein in the presence or absence of amantadine. Cells harboring pColdII(sp-4) <i>m2</i> were cultured as described previously until O.D.<sub>600</sub> reached 0.5–0.6 and incubated at 15°C for 45 to 60 mins. 1 mM IPTG was added to at 5-hr and the culture was divided into two, one of which contains 50 μM amantadine. Cells from 1.5 ml culture were collected at 0 hr, 1 hr, 3 hr, 5 hr, 7 hr, and 19 hr after induction and subjected to SDS-PAGE assay.</p

    Inhibition of M2 channel activity in Oocyte assay.

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    <p>The inhibition efficiency of each compound was examined at the concentration of 100 μM. The values in the table represent how much the M2 channel activity is inhibited. WT, wild type M2 channel. V27A, M2 channel with V27 A mutation. The Oocyte assay was conducted as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054070#pone.0054070-Balannik2" target="_blank">[12]</a>.</p

    Screening for the inhibitors of M2 proton channel.

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    <p>(<b>A</b>) Expression of AcGFP-M2 protein in the presence of a group of compounds to be screened. Experiment procedure is the same as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054070#pone-0054070-g001" target="_blank">Figure 1A</a>. Lane 1, 1 mM IPTG is added to the culture at 0 hr, Lane 2, compound 10, Lane 3, compound 15, Lane 4, compound 34, Lane 5, compound 35, Lane 6, compound 180, Lane 7, compound 189, Lane 8, compound 206, Lane 9, compound 282, Lane 10, compound 293, Lane 11, compound 314, Lane 12, compound 332, Lane 13, compound 343, Lane 14, compound 352, Lane 15, compound 360, Lane 16, compound 360, Lane 17, amantadine, Lane 18, control without any additional compounds. (<b>C</b>) Expression of AcGFP-M2 (V27A) fusion protein in the presence of a group of compounds to be screened as described in (A). Positions of AcGFP-M2 fusion protein are indicated by arrowheads. (<b>B</b>) and (<b>D</b>), Cell density was measured as OD<sub>600</sub> of each overnight culture that expressing AcGFP-M2 fusion protein, and plotted as histogram corresponding to the compounds added. (<b>E</b>) After adding 1 mM IPTG for induction of AcGFP-M2 fusion protein, 100 μl of cell culture was loaded to a 96-well plate, in which a certain compound is added to each well (n = 3). OD<sub>600</sub> was collected after overnight incubation. All the compounds' concentration was 50 μM.</p

    Expression of M2 protein in the SPP system.

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    <p>(<b>A</b>) E. coli cells harboring pColdII(sp-4) <i>m2</i> (from residue 2 to 49 of M2 protein) and pACYC<i>mazF</i> were grown at 37°C to OD<sub>600</sub> = 0.5∼0.6, followed by cold-shock at 15°C for about 60 min. 1 mM of IPTG was added at 0 hr (Lane 1) for induction of M2 protein and MazF. Expression of M2 protein in the SPP system was examined in the presence of amantadine at different concentrations. Lane 2, 0 μM; Lane 3, 50 μM; Lane 4, 100 μM; Lane 5, 200 μM. After overnight incubation for 19 hours, cells from each culture were collected and subjected to SDS-PAGE. (<b>B</b>) Expression of M2 protein in the presence of other compounds besides amantadine. The final concentration of each compound in the culture is 50 μM. The experiments were carried out as described in (A). Lane 1, 1 mM IPTG is added to the culture at 0 hr, Lane 2: C, control without any additional compounds. Lane 3, compound 10, Lane 4, compound 15, Lane 5, compound 34, Lane 6, compound 35, Lane 7, compound 282, Lane 8, compound 293, Lane 9, compound 314, Lane 10, A, amantadine. (<b>C</b>) Expression of AcGFP-M2 fusion protein in the SPP system was carried out as described in (B). Positions of M2 protein and AcGFP-M2 fusion protein are indicated by arrowheads. (<b>D</b>) Cell density was measured as OD<sub>600</sub> of each overnight culture that expressing AcGFP-M2 fusion protein, and plotted as histogram corresponding to the compounds added. (<b>E</b>) Growth curve of cultures to express M2 or AcGFP-M2 fusion protein. Cultures were started at 0 hr and the following experiment procedures are similar to that described in (A). OD <sub>600</sub> of each culture is measured at every time point. M2 protein was induced at 5 hr with (▪) or without (•) amantadine. AcGFP-M2 fusion protein was induced at 5 hr with (♦) or without amantadine (▴).</p

    Novel and Environmentally Friendly Oil Spill Dispersant Based on the Synergy of Biopolymer Xanthan Gum and Silica Nanoparticles

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    The potential toxicity of existing chemical dispersants on the marine environment has motivated the search for environmentally friendly dispersants with excellent dispersion ability. Here, an effective Pickering emulsifier is developed based on the synergy of natural biopolymer, Xanthan Gum (XG), and silica nanoparticles. The oil–in–seawater emulsion stabilized by a combination of XG and silica demonstrates great stability and smaller droplet size, which is favorable for the following natural degradation of oil. The synergistic emulsification mechanism has been investigated systematically. The presence of XG favors the adsorption of silica nanoparticles at the oil–seawater interface and also is considerably effective in enhancing the viscosity of continuous phase. These contributions of XG slow down the droplet coalescence and creaming significantly. Confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) images of emulsions indicate a thick layer of aggregated XG/silica particles at the oil–water interface. This thick layer provides an effective steric barrier. In this study, the synergy between XG and silica not only enhances the dispersion effectiveness, but also reduces the amount of nanoparticles dramatically. This finding opens up a new path for the development of a novel, high efficiency, ecologically acceptable, and cheaper dispersant for emulsifying crude oil following a spill

    Preparation of Oil-in-Seawater Emulsions Based on Environmentally Benign Nanoparticles and Biosurfactant for Oil Spill Remediation

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    One remediation technique of oil spills is the application of dispersants to oil slicks, which is essentially a process of emulsification. Tetradecane and crude oil-in-seawater emulsions formed with silica nanoparticles modified <i>in situ</i> with rhamnolipid produced a longer stability and smaller droplet size. The interactions of silica particles with rhamnolipid were characterized by contact angle, interfacial tension, TEM, and SEM measurements. The images of confocal fluorescence microscopy and SEM showed the oil droplet microstructure and the morphology of nanoparticles at the oil droplet–water interface. The average emulsion droplet size and emulsion index were investigated. These results indicated a synergistic stabilization upon rhamnolipid addition. The synergy was even more efficient in the case of seawater with a high salinity. Here, because of the strong flocculation caused by high salinity, silica nanoparticles alone were not an effective emulsifier in seawater. The modification of silica nanoparticles by rhamnolipid changed the contact angle and promoted their adsorption at the oil–seawater interface, which provided an efficient barrier to droplet coalescence. The emulsification of rhamnolipid-modified silica nanoparticles worked well in crude oil–seawater system. So, this could be a new method to deal with the issue of the marine oil spill by environmentally benign silica particles and rhamnolipid
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