Melting curve analysis was conducted

over a range of 55 to 95°C to assess specificity of amplification. Interleukin-8 expression was normalized to the housekeeper gene, C1orf33, and fold changes in expression relative to the sterile-broth control was calculated using the 2-ΔΔCT method. Statistical analysis Experiments were conducted at least three times on separate occasions https://www.selleckchem.com/products/poziotinib-hm781-36b.html (i.e., replicates). Each assay was conducted at least in duplicate (i.e., observations), and the mean value was used for analysis. Data are expressed as mean ± SEM. All statistical calculations were performed with GraphPad InStat v.3.06 software (GraphPad Software Inc., San Diego, CA). Data with three or more treatments were compared by one way analysis R428 of Adriamycin clinical trial variance, followed by the protected Tukey-Kramer multiple comparison test. Data with two treatments were compared using an unpaired Student’s t-test. Regression analysis was performed using Pearson correlation analysis. Statistical

significance was established at P < 0.05. Acknowledgements We thank Jenny Gusse for conducting the AFLP genotyping and cluster analysis, sequencing the 16S rRNA gene, and for designing and validating the C. concisus-specific cpn60 primers. We also thank Kathaleen House for isolating and conducting the initial characterization of C. concisus isolates. We wish to thank the anonymous reviewers of this manuscript for their insightful and constructive comments. This work was supported by a Peer Review Grant from Agriculture and Agri-Food Canada (Growing Forward initiative). Electronic supplementary material Additional file

1: Dendrogram of C. concisus AFLP profiles demonstrating reproducibility between duplicate independently-prepared samples. AFLP profiles were derived using the unweighted-pair group average linkage of Pearson-product-moment correlation coefficients from 22 Campylobacter Glycogen branching enzyme concisus fecal isolates (designated CHRB) and the type strain (LMG7788). The bar indicates percentage similarity. *, isolates for which only a single profile was analyzed. Additional file 1 contains a figure. (JPEG 111 KB) Additional file 2: Transepithelial resistance (TER) and FITC-dextran permeability for confluent, polarized T84 monolayers inoculated with Campylobacter concisus isolates a . Additional file 2 contains a table. (DOC 24 KB) Additional file 3: PCR screening of genes coding for cytolethal distending toxin (CDT), zonula occludens toxin (Zot), and S-layer RTX for Campylobacter concisus isolates. Additional file 3 contains a table. (DOC 22 KB) References 1. Aabenhus R, On SL, Siemer BL, Permin H, Andersen LP: Delineation of Campylobacter concisus genomospecies by amplified fragment length polymorphism analysis and correlation of results with clinical data. J Clin Microbiol 2005,43(10):5091–5096.PubMedCrossRef 2.