Differences in plasma concentrations between patients suffering from periodontitis (N = 80) and age-matched unaffected controls (N = 48).

<p>Boxes refer to the 25^th (bottom) and 75^th (up) percentiles and the median is the large horizontal line, fences refer to the 10^th (lower) and 90^th (upper) percentiles respectively. Open circles represent outliers whereas asterisks stand for extreme observations with the subject number. Statistical differences are computed with Mann-Whitney U-test.</p

Mean values (standard errors) and mean changes (standard errors) (compared to baseline) of HSP10 (A–B), BiP (C–D) and HSP60 (E–F) before, 24 hrs after and 6 months after periodontal therapy.

<p>Positive differences indicate a relative increase in plasma concentrations of various markers compared to the pre-treatment baseline. Subjects who received intensive periodontal therapy (open circles, N = 40) showed greater plasma concentrations of HSP10 at each time visit when compared to control therapy group subjects (filled circles, N = 40). No other changes were observed. Asterisks refer to statistically significant difference (P<0.05) between groups as computed with analysis of covariance.</p

Case control study

*<p>Differences were computed with parametric (Independent t-test) or non parametric (Mann-Whitney U-Test) dependent upon the data frequency distribution. Categorical variables were analyzed by Chi-square testing.</p>**<p>See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001198#pone-0001198-g002" target="_blank">Figure 2</a>.</p

Binding of DNA to ComE1.

<p>(A) Inhibition ELISA to determine the capacity of Fn or genomic DNA isolated from <i>P. multocida</i> to block the binding of rGST-ComE1 from <i>P. multocida</i> to immobilised human serum Fn. The competing ligands were added at concentrations ranging from 1 mg/ml to 0.1 µg/ml. (B) Inhibition ELISA to determine the capacity of Fn or pUC19 DNA to block the binding of rGST-ComE1 from <i>P. multocida</i> to immobilised human serum Fn. Equimolar concentrations of Fn of pUC19 DNA were used to compete with immobilised Fn for binding to rGST-ComE1. Results are presented as the mean±SEM of quadruplicate wells and are representative of at least three experiments. The values for rGST-ComE1 refer to the binding of this protein to Fn-coated wells with no competing ligands.</p

K_D values for the interaction of recombinant ComE1 proteins with pUC19.

<p>K_D values for the interaction of recombinant ComE1 proteins with pUC19.</p

Binding of wild-type <i>A. pleuropneumoniae</i> or <i>ApΔcomE1</i> to Fn or pUC19 DNA.

<p>Bacteria were grown to early exponential phase (A), late exponential phase (B) or stationary phase (C) and tested for their ability to bind to wells coated with Fn or DNA. Asterisks indicate significant differences (independent t-test; see text for P values) in the binding of wild-type <i>A. pleuropneumoniae and ApΔcomE1</i> to each ligand. Restoration of binding of stationary phase <i>ApΔcomE1</i> to wells coated with Fn by provision of the <i>comE1</i> gene on plasmid pMIDGE311 (D).</p

Comparison of the transformation frequency of wild-type <i>A. pleuropneumoniae</i> and <i>ApcomE1</i>.

<p>The effect of the presence of Fn on the transformation frequency of wild-type <i>A. pleuropneumoniae</i> was also tested. Bars with different letters are significantly different from each other (one-way ANOVA, P<0.001).</p

Oligonucleotides used to amplify genes encoding homologues of ComE1. Restriction sites are underlined.

<p>Oligonucleotides used to amplify genes encoding homologues of ComE1. Restriction sites are underlined.</p

Binding of homologs of ComE1 from <i>P. multocida</i> (PM1665), <i>H. influenzae</i> (Hi1008), <i>A. actinomycetemcomitans</i> (Aa1426), <i>A. pleuropneumoniae</i> (APL_1406), <i>M. haemolytica</i> (MhORF35) or <i>M. succiniproducens</i> (Ms0826) expressed as GST-fusions to pUC19 DNA (A) or Fn (B) measured by direct binding ELISA.

<p>Optical density values at 492 nm were converted to estimates of the concentration of bound protein by reference to a standard curve for each protein.</p

Binding of <i>P. multocida</i> to Fn or pUC19 pre-incubated with rGST-ComE1 from <i>P. multocida</i>.

<p>Values are expressed as percentage values of the binding to Fn. Bars with different letters are significantly different from each other (one-way ANOVA, P<0.001). Data are the mean±SEM of triplicate wells. Results are representative of three separate experiments.</p