Pyocyanin, a virulence factor produced by Pseudomonas aeruginosa, has many damaging effects on mammalian cells. Several lines of evidence suggest that this damage is primarily mediated by its ability to generate ROS and deplete host antioxidant defence mechanisms. However, a causal role for oxidative stress has not yet been demonstrated conclusively. Parallel measures of ROS production, antioxidant levels and cytotoxicity provide convincing evidence that pyocyanin-induced cytotoxicity in A549 respiratory cells is mediated by acute ROS production and subsequent oxidative stress. Pyocyanin increased ROS levels in A549 cells as measured by the fluorescent H2O2 probes Amplex Red and DCFH-DA. These effects were attenuated by the antioxidant N-acetylcysteine. Furthermore, pyocyanin-induced depletion of intracellular GSH levels 24 h after exposure was also prevented by pre-treatment of cells with NAC. Under these conditions, NAC protected cells against pyocyanin-induced cytotoxicity as measured by resazurin

Neutrophils undergo rapid constitutive apoptosis that is accelerated following bacterial ingestion as part of effective immunity, but is also accelerated by bacterial exotoxins as a mechanism of immune evasion. The paradigm of pathogen-driven neutrophil apoptosis is exemplified by the Pseudomonas aeruginosa toxic metabolite, pyocyanin. We previously showed pyocyanin dramatically accelerates neutrophil apoptosis both in vitro and in vivo, impairs host defenses, and favors bacterial persistence. In this study, we investigated the mechanisms of pyocyanin-induced neutrophil apoptosis. Pyocyanin induced early lysosomal dysfunction, shown by altered lysosomal pH, within 15 min of exposure. Lysosomal disruption was followed by mitochondrial membrane permeabilization, caspase activation, and destabilization of Mcl-1. Pharmacological inhibitors of a lysosomal protease, cathepsin D (CTSD), abrogated pyocyanin-induced apoptosis, and translocation of CTSD to the cytosol followed pyocyanin treatment and ...

Clearance of neutrophils from inflamed sites is critical for resolution of inflammation, but pathogen-driven neutrophil apoptosis can impair host defenses. We previously showed that pyocyanin, a phenazine toxic metabolite produced by Pseudomonas aeruginosa, accelerates neutrophil apoptosis in vitro. We compared wild-type and pyocyanin-deficient strains of P. aeruginosa in a murine model of acute pneumonia. Intratracheal instillation of either strain of P. aeruginosa caused a rapid increase in bronchoalveolar lavage neutrophil counts up to 18 h after infection. In wild-type infection, neutrophil numbers then declined steadily, whereas neutrophil numbers increased up to 48 h in mice infected with pyocyanin-deficient P. aeruginosa. In keeping with these differences, pyocyanin production was associated with reduced bacterial clearance from the lungs. Neutrophil apoptosis was increased in mice infected with wild-type compared with the phenazine-deficient strain or two further strains that lack ...

Pseudomonas aeruginosa produce the pigment pyocyanin, which is a secondary metabolite and has antibiotic activity. Pyocyanin is a blue-green pigment that is produced by P. aeruginosa in a media that is devoid of iron content. Pyocyanin producing Pseudomonas aerginosa was isolated from soil sample. Pyocyanin was extracted and purified using chloroform and then characterized using TLC, UV-Vis and FTIR scpectroscopy. The Rf value was found to be 0.79. The UV-Vis Spectrum showed a maximum peak at 277 nm. Pyocyanin has a high antimicrobial activity and can act as a broad-spectrum antibiotic. The inhibition concentrations were found to be as low as 5μg/ml. The extracted pyocyanin showed antibacterial and anti-biofilm activities.. ...

Results: The inhibition zone produced by the methanol leaves extract at 500 mg/mL against P. aeruginosa, S. aureus, and S. typhi were 22.8 ± 0.8, 10.7 ± 1.3, and 12.1 ± 1.0 mm, respectively. The MICs and MBCs of the methanol leaves and roots extracts of this plant for P. aeruginosa, S. aureus, and S. typhi were similar, 125 and 250 mg/mL, respectively. The main phenolic compound obtained from R. alveolatus was 1, 2-benzenedicarboxylic acid (89.68%). The methanol leaves extract of R. alveolatus at 62.5 mg/mL prevented biofilm formation by P. aeruginosa and S. aureus and, when applied at 62.5 mg/mL, reduced pyocyanin production in P. aeruginosa by up to 66 percent. The aqueous extracts of R. alveolatus had not antibacterial and anti-quorum sensing activity. ...

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Bacteria synchronize group behaviors using quorum sensing, which is advantageous during an infection to thwart immune cell attack and resist deleterious changes in the environment. In Pseudomonas aeruginosa, the Pseudomonas quinolone signal (Pqs) quorum-sensing system is an important component of an interconnected intercellular communication network. Two alkylquinolones, 2-heptyl-4-quinolone (HHQ) and 2-heptyl-3-hydroxy-4-quinolone (PQS), activate transcriptional regulator PqsR to promote the production of quinolone signals and virulence factors. Our work focused on the most abundant quinolone produced from the Pqs system, 2,4-dihydroxyquinoline (DHQ), which was shown previously to sustain pyocyanin production and antifungal activity of P. aeruginosa. However, little is known about how DHQ affects P. aeruginosa pathogenicity. Using C. elegans as a model for P. aeruginosa infection, we found pqs mutants only able to produce DHQ maintained virulence towards the nematodes similar to wild-type. In addition,

Treatment of the pathogenic bacterium Pseudomonas aeruginosa with sublethal concentrations of diverse antibiotics promotes the accumulation of pyocyanin, an important virulence factor; in turn, pyocyanin confers general tolerance against antibiotics for both P. aeruginosa and other bacteria.

|p|Pyocyanin (PCN) is a blue redox-active secondary metabolite produced by Pseudomonas aeruginosa. PCN is readily recovered in large quantities in sputum from patients patients infected with P. aeruginosa [1]. PCN interferes with multiple cellular functio

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