618 research outputs found

    Immobilization of liposomes on hydrophobically modified polymer gel particles in batch mode interaction

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    ArticleCOLLOIDS AND SURFACES B-BIOINTERFACES. 55(2): 235-240 (2007)journal articl

    Characterization of the binding of botulinum type B 16S toxin to human intestinal epithelial cells

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    Botulinum neurotoxin produced by Clostridium botulinum type B is a complex of 12S and 16S toxins. 12S toxin consists of a neurotoxin and a nontoxic non-HA (NTNH). The 16S toxin consists of a neurotoxin, an NTNH, and a hemagglutinin (HA). Food-borne botulism is caused by these complex toxins, which are ingested orally and absorbed from the digestive tract across the epithelial barrier lining the gut. Here we show that the type B 16S toxin, but not the 12S toxin or the neurotoxin, binds to the T84 human intestinal epithelial cell line. We also demonstrate that the HA moiety in the 16S toxin mediates the toxin binding to the cells. The carbohydrates containing a galactose moiety inhibited the binding of the 16S toxin to the T84 cells, and neuraminidase treatment of the cells increased the 16S toxin binding. The binding of the 16S toxin to the neuraminidase-treated cells was also inhibited by carbohydrates containing a galactose moiety. These results suggest that the type B 16S toxin binds to human intestinal epithelial cells via the galactose moiety in the carbohydrate chain on the cell surface

    Volume reduction of municipal solid wastes contaminated with radioactive cesium by ferrocyanide coprecipitation technique

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    Municipal solid wastes (MSW) with elevated concentrations of radioactive cesium (rad-Cs hereafter) have been generated in some areas of Japan in the aftermath of the Fukushima Daiichi Nuclear Power Plant (F1 hereafter) accident. Both recycling and final disposal of the contaminated MSW have become a difficult problem in the affected areas, resulting in accumulation of treated residues in the treatment facilities. The rad-Cs in MSW, especially fly ash, often showed a high leaching rate. Extraction of contaminated MSW with water or hot oxalic acid followed by selective removal of rad-Cs from the extract using ferrocyanide (Fer hereafter) coprecipitation technique could be an ultimate solution for waste volume reduction. The MSW extracts contain various metal components as well as chelating reagents like oxalic acid, and are often very saline. The composition of the extract varies widely depending on waste sources, applied treatment techniques, and rad-Cs extraction method etc. The applicability of the Fer coprecipitation technique had to be tested and validated before it could be applied for actual treatment. In this work, we applied the Fer technique and observed removal of cesium (Cs) from water and oxalic acid extracts (all spiked with rad-Cs tracer or stable Cs) of various MSW samples collected from uncontaminated areas. Finally, the Fer technique was applied on site for removal of rad-Cs in the extracts of contaminated MSW. By modifying coprecipitation conditions according to solution matrix, Cs removal rates of higher than 95 % could be obtained

    Gas phase characterization of the noncovalent quaternary structure of Cholera toxin and the Cholera toxin B subunit pentamer

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    Cholera toxin (CTx) is an AB5 cytotonic protein that has medical relevance in cholera and as a novel mucosal adjuvant. Here, we report an analysis of the noncovalent homopentameric complex of CTx B chain (CTx B5) using electrospray ionization triple quadrupole mass spectrometry and tandem mass spectrometry and the analysis of the noncovalent hexameric holotoxin usingelectrospray ionization time-of-flight mass spectrometry over a range of pH values that correlate with those encountered by this toxin after cellular uptake. We show that noncovalent interactions within the toxin assemblies were maintained under both acidic and neutral conditions in the gas phase. However, unlike the related Escherichia coli Shiga-like toxin B5 pentamer (SLTx B), the CTx B5 pentamer was stable at low pH, indicating that additional interactions must be present within the latter. Structural comparison of the CTx B monomer interface reveals an additional α-helix that is absent in the SLTx B monomer. In silico energy calculations support interactions between this helix and the adjacent monomer. These data provide insight into the apparent stabilization of CTx B relative to SLTx B

    The Composition of Cosmic Rays at the Knee

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    The observation of a small change in spectral slope, or 'knee' in the fluxes of cosmic rays near energies 10^15 eV has caused much speculation since its discovery over 40 years ago. The origin of this feature remains unknown. A small workshop to review some modern experimental measurements of this region was held at the Adler Planetarium in Chicago, USA in June 2000. This paper summarizes the results presented at this workshop and the discussion of their interpretation in the context of hadronic models of atmospheric airshowers.Comment: 36 pages, 10 figure

    RNA polymerase II stalling promotes nucleosome occlusion and pTEFb recruitment to drive immortalization by Epstein-Barr virus

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    Epstein-Barr virus (EBV) immortalizes resting B-cells and is a key etiologic agent in the development of numerous cancers. The essential EBV-encoded protein EBNA 2 activates the viral C promoter (Cp) producing a message of ~120 kb that is differentially spliced to encode all EBNAs required for immortalization. We have previously shown that EBNA 2-activated transcription is dependent on the activity of the RNA polymerase II (pol II) C-terminal domain (CTD) kinase pTEFb (CDK9/cyclin T1). We now demonstrate that Cp, in contrast to two shorter EBNA 2-activated viral genes (LMP 1 and 2A), displays high levels of promoter-proximally stalled pol II despite being constitutively active. Consistent with pol II stalling, we detect considerable pausing complex (NELF/DSIF) association with Cp. Significantly, we observe substantial Cp-specific pTEFb recruitment that stimulates high-level pol II CTD serine 2 phosphorylation at distal regions (up to +75 kb), promoting elongation. We reveal that Cp-specific pol II accumulation is directed by DNA sequences unfavourable for nucleosome assembly that increase TBP access and pol II recruitment. Stalled pol II then maintains Cp nucleosome depletion. Our data indicate that pTEFb is recruited to Cp by the bromodomain protein Brd4, with polymerase stalling facilitating stable association of pTEFb. The Brd4 inhibitor JQ1 and the pTEFb inhibitors DRB and Flavopiridol significantly reduce Cp, but not LMP1 transcript production indicating that Brd4 and pTEFb are required for Cp transcription. Taken together our data indicate that pol II stalling at Cp promotes transcription of essential immortalizing genes during EBV infection by (i) preventing promoter-proximal nucleosome assembly and ii) necessitating the recruitment of pTEFb thereby maintaining serine 2 CTD phosphorylation at distal regions
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