Evolved Resistance to a Novel Cationic Peptide Antibiotic Requires High Mutation Supply

Background and Objectives A key strategy for resolving the antibiotic resistance crisis is the development of new drugs with antimicrobial properties. The engineered cationic antimicrobial peptide WLBU2 (also known as PLG0206) is a promising broad-spectrum antimicrobial compound that has completed Phase I clinical studies. It has activity against Gram-negative and Gram-positive bacteria including infections associated with biofilm. No definitive mechanisms of resistance to WLBU2 have been identified. Methodology Here, we used experimental evolution under different levels of mutation supply and whole genome sequencing (WGS) to detect the genetic pathways and probable mechanisms of resistance to this peptide. We propagated populations of wild-type and hypermutator Pseudomonas aeruginosa in the presence of WLBU2 and performed WGS of evolved populations and clones. Results Populations that survived WLBU2 treatment acquired a minimum of two mutations, making the acquisition of resistance more difficult than for most antibiotics, which can be tolerated by mutation of a single target. Major targets of resistance to WLBU2 included the orfN and pmrB genes, previously described to confer resistance to other cationic peptides. More surprisingly, mutations that increase aggregation such as the wsp pathway were also selected despite the ability of WLBU2 to kill cells growing in a biofilm. Conclusions and implications The results show how experimental evolution and WGS can identify genetic targets and actions of new antimicrobial compounds and predict pathways to resistance of new antibiotics in clinical practice

Linking the Cu(II/I) potential to the onset of dynamic phenomena at corroding copper microelectrodes immersed in aqueous 0.5 M NaCl

Electrochemical studies have been conducted at copper microelectrodes (125, 50, and 25 μm in diameter) immersed in aqueous 0.5 M NaCl. Cyclic and linear sweep voltammetry were used to explore the corrosion of copper in chloride media. Cyclic voltammetry revealed the reversible Cu(I)/Cu(0) potential at approximately −0.11 V vs. SCE associated with the formation of a dense CuCl blocking layer (confirmed by in situ Raman and fluorescence measurements). Although continuous dissolution of Cu(I) occurs, only an increase in the driving potential into the region of the Cu(II)/Cu(I) potential at approximately +0.14 V vs. SCE started more rapid and stochastic dissolution/corrosion processes. The corrosion process is demonstrated to be linked to two distinct mechanisms based on (A) slow molecular dissolution and (B) fast colloidal dissolution. A polymer of intrinsic microporosity (PIM-EA-TB) is employed to suppress colloidal processes to reveal the underlying molecular processes.</p

Multimessenger observations of a flaring blazar coincident with high-energy neutrino IceCube-170922A

Previous detections of individual astrophysical sources of neutrinos are limited to the Sun and the supernova 1987A, whereas the origins of the diffuse flux of high-energy cosmic neutrinos remain unidentified. On 22 September 2017, we detected a high-energy neutrino, IceCube-170922A, with an energy of e290 tera-electron volts. Its arrival direction was consistent with the location of a known g-ray blazar, TXS 0506+056, observed to be in a flaring state. An extensive multiwavelength campaign followed, ranging from radio frequencies to g-rays. These observations characterize the variability and energetics of the blazar and include the detection of TXS 0506+056 in very-high-energy g-rays. This observation of a neutrino in spatial coincidence with a g-ray-emitting blazar during an active phase suggests that blazars may be a source of high-energy neutrinos