Synergic action of thermosensitive hydrogel and Au/Ag nanoalloy for sensitive and selective detection of pyocyanin.

The rapid detection of bacterial strains has become a major topic thoroughly discussed across the biomedical field. Paired with the existence of nosocomial pathogen agents that imply extreme medical and financial challenges throughout diagnosis and treatment, the development of rapid and easy-to-use sensing devices has gained an increased amount of attention. Moreover, antibiotic resistance considered by World Health Organization as one of the "biggest threats to global health, food security, and development today" enables this topic as high priority. Pseudomonas aeruginosa, one of the most ubiquitous bacterial strains, has various quorum-sensing systems that are a direct cause of their virulence. One of them is represented by pyocyanin, a blue pigment with electroactive properties that is synthesized from early stages of bacterial colonization. Thus, the sensitive detection of this biomarker could enable a personalized and efficient therapy. It was achieved with the development of an electrochemical sensor based on a thermosensitive polymer, modified with Au/Ag nanoalloy for the rapid and accurate detection of pyocyanin, a virulence biomarker of Pseudomonas aeruginosa. The sensor displayed a linear range from 0.12 to 25 µM, and a limit of detection of 0.04 µM (signal/noise = 3). It was successfully tested in real samples spiked with the target analyte without any pretreatment other than a dilution step. The detection of pyocyanin with high recovery in whole blood in a time frame of 5-10 min from the moment of collection was performed with this electrochemical sensor. Graphical abstract.

Pyocyanin induces systemic oxidative stress, inflammation and behavioral changes in vivo.

Pyocyanin (PCN) is a virulence factor secreted by Pseudomonas aeruginosa (P. aeruginosa) that has been shown to have numerous toxic effects in both in vitro and in vivo studies. Such toxicities include pro-inflammatory and pro-oxidant mediated responses. It is hypothesized that PCN can cross biological membranes and reach the systemic circulation, but no previous studies have investigated this. The aim of this study was, therefore, to quantify PCN in plasma and assess if systemic responses were occurring after localized intranasal administration in C57BL/6 J mice. This was achieved through the plasma quantification of PCN and assessment of changes to behavior using two commonly used tests, the forced swimming test and the open field test. Furthermore, evidence of systemic oxidative stress and inflammation was measured using malondialdehyde (MDA) and TNF-α post PCN exposure. PCN was found to cross into systemic circulation but in a variable manner. Furthermore, significant increases in plasma TNF-α and MDA (both p < 0.001) were observed along with changes in behavior indicative of systemic inflammatory responses.