05). A post-hoc, all pairwise multiple comparison procedure (Tukey Test) was performed for statistical analysis of significance. Tissue cytokine transcript analysis Files from the Luminex® and Open® Array analyses were parsed and organized into tab-delimited files using custom perl scripts. Values across multiple days and sexes were averaged to result in one value for each of 6 experimental conditions (Control, L-MAP, K-MAP, L-NP-51, K-MAP + L-NP-51 and L-MAP + L-NP-51). Targets (cytokines or transcripts)

that gave reliable results above background were included in the final analysis. All values were normalized to control values and expressed as log base 2. Gut microbiota analysis For microbiota analysis, .sff files generated from 454 sequencing were demultiplexed, converted to .fastq files and resulting sequences were trimmed and mapped to 16S ribosomal DNA Selleck Sirolimus intergenic regions to classify the origin

of the sequence. The methodology associated with 454 sequencing were conducted by Research and Laboratory Testing (Lubbock, TX) according to protocols previously developed and described by Dowd et al., [44]. Sequencing data were deposited to GenBank short reads archive (SRA056455). The percent of sequences Bioactive Compound Library clinical trial from each organism in each sample was normalized across all samples and final values were normalized to control and values were expressed as log base 2 of the difference between each sample and the control. A custom R script was written to perform a Pearson correlation between the relative abundance of each genus and relative abundance of each cytokine; geni with p-values of <0.05 in the Pearson and at least one cytokine from the Luminex® analysis were included in the final table, separated based on whether the r-value was positive (positive correlation) or negative

(negative correlation). Acknowledgements We would like to thank Nutrition Physiology Incorporated (NPC) and the Centers of Excellence support for mafosfamide the International Center for Food Industry Excellence for their contributions towards this study, including Dr. Doug Ware from NPC. We would also like to thank the TTU Core Facility and TTU Molecular Pathology Program for their assistance and contributions. Additionally, the authors would like to thank the TTU/HHMI Undergraduate Research Program for their support of David Campos. We would like to thank Dr. Judith Stabel at the NADC and Drs. Mohamed Osman and Don Beitz at ISU for their contributions. Funding Nutrition Physiology Incorporated provided funding for this study, including some salary for Mindy M. Brashears, Enusha Karunasena, Estevan Kiernan, Russell Lackey, and Paresh Kurkure. References 1.