245 research outputs found

    Genomics-based approaches used in the control of EIDs from the outbreak of a disease to the development of a vaccine or drug.

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    <p>(A) The causative agent of a disease may first be identified from patient samples by using metagenomics. (B) Vaccine and therapeutic targets can be identified from the pathogen genome using a variety of screening approaches that focus on the genome, transcriptome, proteome, immunome or structural genome. (C) The human genome can be screened to avoid homologies or similarities with pathogen vaccine and therapeutic targets, or to identify new targets. (D) Once candidate vaccine and therapeutic targets have been identified they must be shown to provide protection against disease and to be safe for use in patients. (E) The clinically tested vaccine or therapeutic can then be licensed for use. The clinical responses of a vaccine and/or therapeutic can be analyzed using human genome based studies (dotted arrows). The pathogen genome can also be used to analyze mutants that are able to evade the immune system in vaccinated subjects or organisms that develop antibiotic resistance. Examples of the approaches indicated are given in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000612#pgen-1000612-t001" target="_blank">Table 1</a>.</p

    Concentration of cytokine produced by AB2.2 mouse ESCs or ESDCs, alone and during infection with <i>S.</i> Typhimurium SL1344(p1C/1).

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    <p>Each value represents the average of at least two measurements ± average deviation. The detection limits of the assay were 2.5 pg/ml for IFNγ, 17.5 pg/ml for IL-10, 10.7 pg/ml for IL-12p70, 5 pg/ml for IL-6, 52.5 pg/ml for MCP-1 and 7.3 pg/ml for TNFα values below these are listed as 0 in the table).The data was analyzed using one-way ANOVA,</p>*<p>indicates a significant difference with p<0.05.</p

    Innate DB pathways up-regulated by ESC derived DCs at 4 hours post-infection.

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    *<p>Interaction Node;</p>1<p>S-Phase Network;</p>2<p>p53-Independent DNA Damage Response Network;</p>3<p>Host interactions of HIV factors Network;</p>4<p>Signaling by Wnt Network;</p>5<p>DNA Replication.</p

    Overview of the <i>viaB</i> operon and Vi capsular polysaccharide biosynthesis pathway.

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    <p>(<b>A</b>) Schematic diagram of the <i>S</i>. Typhi <i>viaB</i> operon. (<b>B</b>) The Vi antigen is a linear polymer of α-1,4-linked <i>N</i>-acetylgalatosaminuronate, nonstoichiometrically esterified with acetyl groups at the C-3. A TviB-catalyzed oxidation of UDP-<i>N</i>-acetylglucosamine (UDP-GlcNAc) followed by the TviC-catalyzed epimerization at the C-4 of UDP-<i>N</i>-acetylglucosaminuronate (UDP-GlcNAcA) results in the formation of UDP-<i>N</i>-acetylgalactosaminuronate (UDP-GalNAcA), the building block for Vi polymer formation. The Vi polymer is synthesized in the cytoplasm and assembly is dependent on TviD and TviE. The enzyme catalyzing the O-acetylation of the capsular polysaccharide has not yet been identified. Subsequent translocation of the polysaccharide to the cell surface follows an ATP-binding cassette (ABC) transporter-dependent process. The transporter consists of VexA, VexB, VexC and VexD. The precise function of VexE is equivocal although it might be involved in anchoring the Vi to the cell surface.</p

    Electron microscope observations revealed details of internal organelles of undifferentiated and differentiated cells.

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    <p>Electron micrographs showing (A) undifferentiated ESC and (B) ESDC. Note the presence of intracellular <i>S.</i> Typhimurium in these images. Scale bars denote different sizes.</p

    Gentamicin invasion assay.

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    <p>The ability of <i>S.</i> Typhimurium SL1344(p1C/1) to invade ESDCs was measured by the gentamicin assay described in Methods. ESDCs were exposed to either wild type <i>S</i>. Typhimurium or a <i>SipB</i> mutant for 30 minutes and then incubated with media containing 50 µg/ml of Gentamicin for 2 or 4 hours and then invading bacteria were enumerated.</p

    Graphic representation of the quantified expression of chosen genes versus un-infected ESDCs.

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    <p>Total RNA extracted from infected ESDCs was used in semi-quantitative Real Time-PCR to confirm the BioConductor analysis of up- or down-regulated genes. The data were analyzed using the ΔΔct value calculated versus the un-infected ESDCs.</p
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