For this analysis, all men who were non-compliant to study drug were removed

600 of CV026., P<0.05 compared with the control. , P<0.001 compared with the control. doi:10.1371/journal.pone.0135495.g002 Effect of Cinnamon Oil on P. aeruginosa Virulence Factors P. aeruginosa PAO1 produces a green-colored cytotoxic factor that plays an important role during P. aeruginosa host infections. Genes responsible for the synthesis and secretion of the pyocyanin pigment are regulated by QS. The effects of different concentrations of cinnamon oil on the ability of the bacteria to synthesize and secrete pyocyanin were investigated. The results indicated that the extent of pyocyanin formation decreased with increasing concentrations of cinnamon oil, showing a reduction to approximately 22% at a concentration of 0.2 l/ ml compared with the Relebactam chemical information control, as shown in Fig 4. Swarming usually occurs on semi-solid agar. In this study, 0.5% swarming agar was optimized. The swarming motility of P. aeruginosa PAO1 was evaluated by spotting a culture of P. aeruginosa PAO1 on swarming agar plates with or without cinnamon oil and measuring the lengths of dendrites on these plates. Swarming inhibition against P. aeruginosa PAO1 could be observed on swarming agar containing 0.2 l/ml cinnamon oil. Alginate is one of the major constituents of the EPS secreted by P. aeruginosa PAO1 during biofilm formation. The efficacy of cinnamon oil in reducing alginate production was examined, and alginate production was found to be reduced significantly with increasing concentrations 7 / 18 Cinnamon Oil Inhibits Pseudomonas aeruginosa Quorum-Sensing Fig 3. Effects of different concentrations of cinnamon oil on the concentration of 3-oxo-C12HSL produced by P. aeruginosa PAO1grown up to 24h. doi:10.1371/journal.pone.0135495.g003 of cinnamon oil. There was a 54% reduction in alginate production at a concentration of 0.2 l/ ml in comparison with the control. Total protease activity was measured using two assays: the azocasein assay and the skimmed milk agar plate assay. In the azocasein assay, the protease activities of untreated cultures and cultures grown in the presence of increasing concentrations of oil were estimated. A decrease in protease activity of approximately 22% was observed after treatment with 0.2 l/ml cinnamon oil compared with the control, and there was a further decrease in protease production with an increase in the concentration of oil, possibly due to the combined effect of a reduced growth rate and QS inhibition. In the skim milk agar assay, the supernatants from treated PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19723632 and untreated cultures were added to the wells of a skim milk agar plate, and the clearance zones were shown to decrease with higher concentrations of oil. 8 / 18 Cinnamon Oil Inhibits Pseudomonas aeruginosa Quorum-Sensing Fig 4. Effects of different concentration of cinnamon oil on the production of pyocyanin. Error bars indicate the standard deviations for three measurements, and the data were normalized according to the OD600., P<0.05 compared with PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19723666 the control. doi:10.1371/journal.pone.0135495.g004 Effect of Cinnamon oil on Biofilm Formation Biofilm quantification was performed to investigate the effect of cinnamon oil on biofilm formation. In a conventional crystal violet binding assay, biofilm formation was observed to be reduced by approximately 31% in the presence of 0.2 l/ml cinnamon oil compared with the control. Microscopy Analysis of Biofilms To clearly demonstrate the effect of cinnamon oil on biofilm, microscopic examinations were performed in the presence of hig