Figure 8 Hypothetical model of buy AR-13324 isolimonic action on EHEC. The isolimonic acid seems to modulate the AI-3/Epinephrine mediated signaling in QseBC and QseA dependent manner. Broken arrow indicate unknown mode of interaction of AI-3 with qseA. Wavy arrows
indicate interaction of isolimonic acid with qseBC and qseA is unknown. Conclusion The present study demonstrates that the citrus limonoids, particularly isolimonic acid and ichangin are strong inhibitors of biofilm formation and attachment of EHEC to Caco-2 cells. Furthermore, isolimonic acid and ichangin seems to affect biofilm formation and TTSS by repressing LEE and flagellar operon. Isolimonic acid seems to exert its effect by inhibiting AI-3/epinephrine mediated cell-cell signaling in QseBC and QseA dependent manner. However, the mechanism this website by which isolimonic acid affects the QseBC and QseA remains to be elucidated. One possibility is that the isolimonic acid may interfere with the DNA binding activities of QseB and QseA. Another possible scenario will be that isolimonic acid interferes Selleck XAV-939 with phosphorylation events. However, further study is required to determine the target of isolimonic acid for the modulation of flhDC and ler. In addition, determination of the structure-activity relationship
will provide further insights into the inhibitory action of isolimonic acid. In nutshell, isolimonic acid acts as an antivirulence agent in EHEC and may serve as lead compound for development of novel agents. Furthermore, the fact that isolimonic acid is present in citrus juices and do not demonstrate cytotoxic effect on normal human cell line
, increases the desirability to develop it as antivirulence agent. Acknowledgements We would like to thank Dr. V. Sperandio (University of Texas Southwestern Medical Center, Dallas, TX) for generously providing AI-3 reporter strains harboring chromosomal LEE1:lacZ (TEVS232), LEE2:lacZ (TEVS21) and EHEC mutants VS145, VS151, VS138, VS179. This project is based upon the work supported by the USDA-NIFA No. 2010-34402-20875, “Designing Foods for Health” through the Vegetable & Fruit Improvement Center. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. PLEKHM2 Electronic supplementary material Additional file 1: Figure S1: Metabolic activity of E. coli O157:H7 in presence of 100 μg/ml limonoids as measured by AlamarBlue reduction. (DOC 102 KB) References 1. Nataro JP, Kaper JB: Diarrheagenic Escherichia coli . Clin Microbiol Rev 1998,11(1):142–201.PubMed 2. Tarr PI, Gordon CA, Chandler WL: Shiga-toxin-producing Escherichia coli and haemolytic uraemic syndrome. Lancet 2005,365(9464):1073–1086.PubMed 3. Griffin D, Springer D, Moore Z, Njord L, Njord R, Sweat D, Lee N, Maillard J-M, Davies M, Fleischauer A, et al.: Escherichia coli O157:H7 Gastroenteritis Associated with a State Fair — North Carolina, 2011. Morb Mort Weekly Rep 2012,60(51):1745–1746. 4.