Bacteria were stained with acridine orange as described for Panel

Bacteria were stained with acridine orange as described for Panel A, then photographed using a Retiga digital camera. Digital images were captured or converted to black-and-white, then subjected to image analysis using ImageJ, free image analysis software developed at the NIH. The version we used is called Fiji (ImageJ for MacIntosh, version 1.47n). Detailed instructions on how to open and process the files are available from the author at [email protected]. Bacterial lengths were determined for each condition and expressed as a ratio compared to the no- ciprofloxacin, no-metal control bacteria.

Panel C, effect of metals on bacterial elongation in STEC strain Mocetinostat cost Popeye-1, using the same methods described for Panel B. Panel D, effect of zinc on mitomycin C-induced bacterial elongation. In Panel D the actual bacterial length is shown (in micrometers) using 2 micrometer size beads for calibration. (PDF 952 KB) Additional file 2: Table S1: Effects of Biometals at Multiple Phases of STEC and EPEC Pathogenesis. (PDF 96 KB) References 1. Bhutta ZA, Bird SM, Black RE, Brown KH, Gardner JM, Hidayat A, Khatun F, Martorell R, Ninh NX, Penny ME, Rosado JL, Roy SK, Ruel M, Sazawal S, Shankar A: Therapeutic effects of oral zinc in acute and persistent PXD101 concentration diarrhea in children in developing countries: pooled analysis of randomized controlled trials. Am J Clin Nutr 2000, 72:1516–1522.PubMed 2. Sazawal S, Black R,

Bhan M, Bhandari N, Sinha A, Jalla S: Zinc supplementation in young children with acute diarrhea in India. N Engl J Med 1995, 333:839–844.PubMedCrossRef 3. Patel A, Mamtani M, Dibley MJ, Badhoniya N, Kulkarni H: Therapeutic value of zinc supplementation in acute and persistent diarrhea: a systematic review. PLoS One 2010, 5:e10386.PubMedCentralPubMedCrossRef 4. Selleck NVP-HSP990 Gabbianelli R, Scotti R, Ammendola S, Petrarca P, Nicolini L, Battistoni A: Role of ZnuABC and ZinT in Escherichia coli O157:H7 HDAC inhibitor zinc acquisition and interaction with epithelial cells. BMC Microbiol 2011, 11:36.PubMedCentralPubMedCrossRef

5. Porcheron G, Garenaux A, Proulx J, Sabri M, Dozois CM: Iron, copper, zinc, and manganese transport and regulation in pathogenic Enterobacteria: correlations between strains, site of infection and the relative importance of the different metal transport systems for virulence. Front Cell Infect Microbiol 2013, 3:90.PubMedCentralPubMedCrossRef 6. Prasad AS: Zinc: mechanisms of host defense. J Nutr 2007, 137:1345–1349.PubMed 7. Karlsen TH, Sommerfelt H, Klomstad S, Andersen PK, Strand TA, Ulvik RJ, Åhrén C, Grewal HMS: Intestinal and systemic immune responses to an oral cholera toxoid B subunit whole-cell vaccine administered during zinc supplementation. Infect Immun 2003, 71:3909–3913.PubMedCentralPubMedCrossRef 8. Wellinghausen N, Martin M, Rink L: Zinc inhibits interleukin-1-dependent T cell stimulation. Eur J Immunol 1997, 27:2529–2535.PubMedCrossRef 9. Schlesinger L, Arevalo M, Arredondo S, Lonnerdal B, Stekel A: Zinc supplementation impairs monocyte function.

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