BACKGROUND: The objectives of the current study were to extract pyocyanin from Pseudomonas aeruginosa clinical isolates, characterize its chemical nature, and assess its biological activity against different bacteria and cancer cells. Due to its diverse bioactive properties, pyocyanin, being one of the virulence factors of P. aeruginosa, holds a promising, safe, and available therapeutic potential.
METHODS: 30 clinical P. aeruginosa isolates were collected from different sources of infections and identified by routine methods, the VITEK 2 compact system, and 16 S rRNA. The phenazine-modifying genes (phzM, phzS) were identified using polymerase chain reaction (PCR). Pyocyanin chemical characterization included UV-Vis spectrophotometry, Fourier Transform Infra-Red spectroscopy (FTIR), Gas Chromatography-Mass Spectrometry (GC-MS), and Liquid Chromatography-Mass Spectrometry (LC-MS). The biological activity of pyocyanin was explored by determining the MIC values against different clinical bacterial strains and assessing its anticancer activity against A549, MDA-MB-231, and Caco-2 cancer cell lines using cytotoxicity, wound healing and colony forming assays.
RESULTS: All identified isolates harboured at least one of the phzM or phzS genes. The co-presence of both genes was demonstrated in 13 isolates. The UV-VIS absorbance peaks were maxima at 215, 265, 385, and 520 nm. FTIR could identify the characteristic pyocyanin functional groups, whereas both GC-MS and LC-MS elucidated the chemical formula C11H18N2O2, with a molecular weight 210. The quadri-technical analytical approaches confirmed the chemical nature of the extracted pyocyanin. The extract showed broad-spectrum antibacterial activity, with the greatest activity against Bacillus, Staphylococcus, and Streptococcus species (MICs 31.25-125 µg/mL), followed by E. coli isolates (MICs 250-1000 µg/mL). Regarding the anticancer activity, the pyocyanin extract showed IC50 values against A549, MDA-MB-231, and Caco-2 cancer cell lines of 130, 105, and 187.9 µg/mL, respectively. Furthermore, pyocyanin has markedly suppressed colony formation and migratory abilities in these cells.
CONCLUSIONS: The extracted pyocyanin has demonstrated to be a potentially effective candidate against various bacterial infections and cancers. Hence, the current findings could contribute to producing this natural compound easily through an affordable method. Nonetheless, future studies are required to investigate pyocyanin\'s effects in vivo and analyse the results of combining it with other traditional antibiotics or anticancer drugs.

This study investigated electrochemical characteristics of Gram-positive, Nocardiopsis dassonvillei B17 facultative bacterium in bioelectrochemical systems. The results demonstrated that anodic and cathodic reaction rates were catalyzed by this bacterium, especially by utilization of aluminium alloy as a substrate. Cyclic voltammogram results depicted an increase of peak current and surface area through biofilm development, confirming its importance on catalysis of redox reactions. Phenazine derivatives were detected and their electron mediating behavior was evaluated exogenously. A symmetrical redox peak in the range of -59 to -159 mV/SHE was observed in cyclic voltammogram of bacterial solution supplemented with 12 μM phenazine, a result consistent with cyclic voltammogram of a 5-d biofilm, confirming its importance as an electron mediator in extracellular electron transfer. Furthermore, the dependency of bacterial catalysis and polarization potential were studied. These results suggested that B17 biofilm behaved as a biocathode and transferred electrons to bacterial cells through a mechanism associated with electron mediators.

The detection of volatile aliphatic aldehydes is of significance because of their chemical toxicity, physical volatility and widespread applications in chemical industrial processes. In this work, the direct detection of aliphatic aldehydes is tackled using a pillar[5]arene-based fluorescent supramolecular polymer with vaporchromic behavior. Thin films with strong orange-yellow fluorescence are prepared by coating the linear supramolecular polymer on glass sheets. When the thin films are exposed to aliphatic aldehydes with different carbon chain lengths, they can selectively sensing n-butyraldehyde (C4 ) and caprylicaldehyde (C8 ), accompanied by fluorescence quenching, indicating that the supramolecular polymer is a highly selective vapochromic response material for aliphatic aldehydes with long alkyl chains.

A series of dyes based on the phenazine skeleton were synthesized. They differed in the number of conjugated double bonds, the arrangement of aromatic rings (linear and/or angular system), as well as the number and position of nitrogen atoms in the molecule. These compounds were investigated as potential singlet oxygen sensitizers and visible light absorbers in dye photoinitiating systems for radical polymerization. The quantum yield of the singlet oxygen formation was determined by the comparative method based on the 1H NMR spectra recorded for the tested dyes in the presence of 2,3-diphenyl-p-dioxene before and after irradiation. The quantum yield of the triplet state formation was estimated based on the transient absorption spectra recorded using the nanosecond flash photolysis technique. The effectiveness of the dye photoinitiating system was characterized by the initial rate of trimethylolpropane triacrylate (TMPTA) polymerization. In the investigated photoinitiating systems, the sensitizer was an electron acceptor, whereas the co-initiator was an electron donor. The effectiveness of TMPTA photoinitiated polymerization clearly depended on the arrangement of aromatic rings and the number of nitrogen atoms in the modified phenazine structure as well as the quantum yield of the triplet state formation of the photosensitizer in the visible light region.

Pyocyanin, a redox-active phenazine pigment produced by Pseudomonas aeruginosa, inhibits 5-lipoxygenase (5-LOX) activity. However, whether pyocyanin can directly block the enzymatic activity by binding at the active site still remains a question because of its ability to produce superoxide radicals and H2O2. With the objective of characterizing this mechanism, we carried out molecular docking and molecular dynamics simulations and performed Molecular Mechanics Poisson-Boltzmann surface area (MMPBSA) binding energy studies. The results of the study revealed that pyocyanin is dynamically stable at the active site of 5-LOX and its MMPBSA binding energy (-84.720 kJ/mol) is comparable to that of the 5-LOX standard inhibitor zileuton (-72.729 kJ/mol). Similar studies using three other phenazine derivatives - 1-hydroxyphenazine, phenazine-1-carboxylic acid and phenazine-1-carboxamide - also showed encouraging results. In light of this evidence, we postulate as a proof of concept that pyocyanin and these phenazine derivatives have the potential to inhibit 5-LOX activity by directly binding at the active site and blocking enzymatic catalysis of the substrate. Considering the potential of 5-LOX inhibitors in inflammatory diseases such as asthma and rheumatoid arthritis, the findings of this study open up the exploration of phenazine derivatives in structure-based drug design against 5-LOX. Communicated by Ramaswamy H. Sarma.

High concentrations of nitrous oxide were recovered from partial nitrification treated leachate in a microbial electrolysis cell (MEC) inoculated with a nosZ-deficient strain of Pseudomonas aeruginosa. N2O conversion efficiencies > 90% were achieved when a potential of 0.8 V was applied to the MEC. The ΔnosZ strain was enriched in the 0.8 V MEC, but Achromobacter dominated the non-current control. Nitric oxide reductase genes were highly expressed by ΔnosZ cells growing in the 0.8 V MEC, consistent with enhanced nitrous oxide production rates. Concentrations of phenazine derivatives and transcripts from phenazine biosynthesis genes were also high in the 0.8 V MEC. Phenazine derivatives are known to act as electron shuttles, enhance biofilm formation, and help ward off competitors, thereby increasing the survivability of the ΔnosZ strain in the MEC. These results show that applied current stabilized growth of the ΔnosZ strain in the reactor and allowed it to sustainably generate high concentrations of nitrous oxide.

Three new phenazine derivatives (1-3), along with known compounds (4-7) of saphenic acid derivatives, were isolated from a deep-sea sediment-derived yeast-like fungus Cystobasidium larynigs collected from the Indian Ocean. The structures of the new compounds (1-3) were determined by analysis of spectroscopic data, semi-synthesis and comparison of optical rotation values. All the isolated compounds (1-7), except for 2, showed nitric oxide (NO) production inhibitory effect against lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells without cytotoxicity at concentrations up to 30 μg/mL. This is the first report on the yeast-like fungus Cystobasidium laryngis producing phenazines and anti-inflammatory activity of 1-7 including saphenic acid (4).

Noncovalent functionalization of graphene with organic molecules offers a direct route to multifunctional modification of this nanomaterial, leading to its various possible practical applications. In this work, the structures of hybrids formed by linear heterocyclic compounds such as imidazophenazine (F1) and its derivatives (F2-F4) with graphene and the corresponding interaction energies are studied by using the DFT method. Special attention is paid to the hybrids where the attached molecule is located along the graphene zigzag (GZZ ) and armchair (GAC ) directions. The interaction energies corresponding to the graphene hybrids of the F1-F4 compounds for the two directions are found to be distinct, while tetracene (being a symmetrical molecule) shows a small difference between these binding energies. It is found that the back-side CH3 and CF3 groups have an important influence on the arrangements of F1 derivatives on graphene and on their binding energies. The contribution of the CF3 group to the total binding energy of the F3 molecule with graphene is the largest (3.4 kcal mol(-1) ) (the GZZ direction) while the CH3 group increases this energy of F2 only by 2.0 kcal mol(-1) (the GAC direction). It is shown that replacing the carbons with other atoms or adding a back-side group enables one to vary the polarizability of graphene.