Mechanistic analysis of a synthetic inhibitor of the LasI quorum-sensing signal synthase

Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen responsible for many human infections. LasI is an acyl-homoserine lactone synthase that produces a quorum-sensing (QS) signal that positively regulates numerous P. aeruginosa virulence determinants. The inhibition of the LasI protein is therefore an attractive drug target. In this study, a novel in silico to in vitro complementation was applied to screen thiazolidinedione-type compounds for their ability to inhibit biofilm formation at concentrations not affecting bacterial growth. The compound (z)-5-octylidenethiazolidine-2, 4-dione (TZD-C8) was a strong inhibitor of biofilm formation and chosen for further study. Structural exploration of in silico docking predicted that the compound had high affinity for the LasI activity pocket. The TZD-C8 compound was also predicted to create hydrogen bonds with residues Arg30 and Ile107. Site-directed mutagenesis (SDM) of these two sites demonstrated that TZD-C8 inhibition was abolished in the lasI double mutant PAO-R30D, I107S. In addition, in vitro swarming motility and quorum sensing signal production were affected by TZD-C 8, confirming this compound alters the cell to cell signalling circuitry. Overall, this novel inhibitor of P. aeruginosa quorum sensing shows great promise and validates our mechanistic approach to discovering inhibitors of LuxI-type acyl-homoserine lactone synthases

The nature and fate of natural resins in the geosphere. Part X.^†Structural characteristics of the macromolecular constituents of modern Dammar resin and Class II ambers

<p/> <p>As part of a larger study of the structure and behavior of polyterpenoids in sedimentary systems, the structural characteristics of the macromolecular constituents of Dammar resin and a related Class II amber have been reinvestigated. The conclusions drawn from these analyses are inconsistent with the current widely held "polycadinene" model for the macromolecular structure of these materials. Double bond characteristics observed by one and two dimensional NMR spectroscopy do not match those in the proposed "polycadinene" structure. Based on these observations it is suggested that the proposed "polycadinene" structure for these materials is inadequate and requires revision. Elemental and NMR data also suggest a significant contribution from functionalized monomers.</p

A novel chronic wound biofilm model sustaining coexistence of Pseudomonas aeruginosa

Chronic wounds are a large burden to patients and healthcare systems. Biofilm infections in chronic wounds are crucial factors leading to non‐healing of wounds. It is important to study biofilm in wounds and to develop effective interventions against wound biofilm. This study presents a novel in vitro biofilm model mimicking infected chronic wounds. The novel layered chronic wound biofilm model uses woundlike media and includes both Pseudomonas aeruginosa and Staphylococcus aureus, which have been identified as the most important pathogens in wounds. The model sustains their coexistence for at least 96 h. Microscopy of the model revealed microbial growth in non‐surface attached microcolonies as previously observed in vivo. The model was used to determine log(10)‐reduction for the use of an antimicrobial solution and antimicrobial dressings (containing silver or honey) showing moderate‐to‐low antibiofilm effect, which indicates better concordance with the observed clinical performance of this type of treatment than other widely used standard tests