89 research outputs found

    Electron microscopy observations of <i>Lb</i>. <i>rhamnosus</i> PB12 and its SrtC1-complemented derivatives.

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    <p>Bacterial cells were immunogold-labeled with anti-SpaA serum and gold particles (10 nm). Legend: A, <i>Lb</i>. <i>rhamnosus</i> GG; B, <i>Lb</i>. <i>rhamnosus</i> PB12; C, <i>Lb</i>. <i>rhamnosus</i> PB12 + SrtC1.</p

    Effect of glucose and lactate on cellular ATP content in IPEC-1 cells after 1 h treatment.

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    <p>IPEC-1 cells were grown to confluence (7 d) and treated for 1 h with HBSS-based solutions. Supernatant was removed and ATP content was measured by a luminescence based assay. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p<0.05) differences.</p

    Anthropocentric view of the <i>L. rhamnosus</i> species.

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    <p>The interactions between <i>L. rhamnosus</i> and the human cavities are frequent and occur in various contexts, <i>i.e.</i> consumption of food products (common scenario) or development of bacteremia (rare event). For each niche or isolation source, the strains were grouped according to their geno-phenotype (radar plot). The geno-phenotype is based on the scoring of distinctive genetic and phenotypic traits measured in this study, <i>i.e.</i> gene-content, CRISPR oligotype, bile resistance, pilosotype, sugar group I (dulcitol, D-arabinose and L-fucose), sugar group II (D-saccharose, D-maltose, methyl-α-D-glucopyranoside and D-turanose) and sugar group III(L-rhamnose, L-sorbose, D-ribose and D-lactose). The distinction between the two main geno-phenotypes mostly relies on gene acquisition and loss, point mutations, genetic reorganization that possibly reflect strain adaptation to an ecological niche.</p

    Mucoadhesiveness of recombinant SpaFED-piliated lactococci.

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    <p><i>In vitro</i> mucus-binding assays with normalized (OD600 = 0.5) cultures of recombinant WT (GRS1189) and SpaF pilin-deleted (GRS1126) SpaFED-piliated lactococci, and as well with the vectorless (GRS71) and empty vector (GRS1052) <i>L. lactis</i> NZ900 control strains, were carried out as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113922#s4" target="_blank">Materials and Methods</a>. Triplicate measurements were done for each sample, with three independent experiments having been performed. The standard error of mean (SEM) is shown as error bars. Pairwise differences between the mucus adhesion data for GRS1189 or GRS1226 cells and that for GRS71 cells are deemed extremely significant (<i>P</i>≤0.0001).</p

    Principal component analysis (PCA) (A) and redundancy analysis (RDA) (B) of fecal samples from healthy young children and adults at the phylum-like level.

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    <p>Log transformed data were used for analysis. In PCA, The first two principal components capture 21% (PCA1) and 16% (PCA2) of variance respectively. RDA plot shows the result from supervised PCA, where group assignment of subjects (adults or children) was used as a dependent variable. In RDA, first and second ordination axes are plotted, explaining 13% and 20% of the variance. Separation between children and adults is significant (p = 0.0002, permutation test).</p

    LrpCBA pilus-mediated cellular variation of TLR2-regulated NF-ÎşB responses and endogenous IL-8 production in live and heat-treated HEK-TLR2 cells.

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    <p>Live (-) and heat-treated (+) normalized cultures of <i>L</i>. <i>ruminis</i> cells and the recombinant GRS1224 (WT) and GRS1225 (LrpC-deleted) lactococci (MOI 100) were combined with HEK-TLR2 cells. TLR2-dependent NF-κB activation <b>(A)</b> and endogenous IL-8 production <b>(B)</b> were measured. GRS71 and GRS1052 cells were used as controls and treated similarly. Measurements also taken for DMEM cell-culture medium and a TLR2-agonist lipopeptide (Pam3CSK4; 1 ng/ml) were used as negative and positive controls, respectively. Triplicate measurements were made for both sets of experiments. Limit bars show SEM. Statistical differences for individual pairwise comparisons against the GRS71 control (unheated) are specified as *** = <i>P</i> ≤ 0.001 (highly significant), ** = <i>P</i> ≤ 0.01 (very significant), or <i>ns</i> = <i>P</i> > 0.05 (not significant). Individual pairwise comparisons of data between the heated and unheated samples are indicated as *** = <i>P</i> ≤ 0.001 (highly significant), ** = <i>P</i> ≤ 0.01 (very significant), * = <i>P</i> ≤ 0.05 (significant), or <i>ns</i> = <i>P</i> > 0.05 (not significant).</p

    LrpCBA pilus-mediated cellular adhesion to ECM-related proteins.

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    <p>Normalized (OD600 = 0.5) cultures of <i>L</i>. <i>ruminis</i> cells and the recombinant WT and LrpC-deleted LrpCBA-piliated lactococci (GRS1224 and GRS1225, respectively) were assayed for <i>in vitro</i> binding to collagen type I <b>(A)</b>, type IV <b>(B)</b>, and fibronectin <b>(C)</b>. Vectorless (GRS71) and empty vector (GRS1052) lactococci were included as controls in these experiments. Measurements were normally made in triplicate, with the standard error of mean (SEM) shown by limit bars. Statistical differences for individual pairwise comparisons against the GRS71 control are denoted as *** = <i>P</i> ≤ 0.001 (highly significant) or ** = <i>P</i> ≤ 0.01 (very significant).</p

    RP-HPLC separation profile of muropeptides from LGG digested with mutanolysin (A) and digested with mutanolysin and Msp1 (B).

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    <p>Peak numbers refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031588#pone-0031588-t001" target="_blank">Table 1</a>. DS-di, disaccharide dipeptide (GlcNAc-MurNAc-L-Ala-D-Gln). The schematic structure of LGG peptidoglycan and site of cleavage of Msp1 is also represented. GlcNAc, N-acetylglucosamine; MurNAc, N-acetylmuramic acid.</p

    Phylogenetic tree analysis of the <i>L</i>. <i>ruminis</i> genomes.

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    <p>Unrooted phylogenies of the <i>L</i>. <i>ruminis</i> genomes were based on the multiple sequence alignment of core proteins and constructed with the neighbor-joining tree-building algorithm (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0175541#sec002" target="_blank">Materials and Methods</a> for details). Names of the <i>L</i>. <i>ruminis</i> strains from which each genome is derived are shown. Four distinct phyletic clades were identified based on host-gut source (human, bovine, porcine, or equine) and are indicated accordingly.</p
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