Project description:Several environmental bacteria encode plastic-degrading enzymes, a potential evolutionary response to the rapid introduction of plastic across global ecosystems. Given the widespread use of plastic in healthcare, we hypothesised that clinical bacterial isolates may also degrade plastic, rendering plastic-containing medical devices susceptible to degradation and failure and potentially offering these pathogens a carbon source that could be used to persist in the hospital-built environment. Here, we mined the genomes of prevalent pathogens and identified several enzymes in different pathogens with homology to known plastic-degrading enzymes. Synthesising and expressing a potential plastic-degrading enzyme derived from a Pseudomonas aeruginosa wound isolate in a heterologous host, we were able to demonstrate potent plastic degrading activity. We subsequently found that the original P. aeruginosa clinical isolate could reduce the weight of a medically relevant plastic, polycaprolactone (PCL), by 78% in 7 days, and critically could use it as a sole carbon source to grow. We uncovered a direct link to virulence, demonstrating that encoding a plastic degrading enzyme can significantly enhance biofilm formation and pathogenicity in vivo. We also demonstrate that this augmented biofilm phenotype is conserved in another P. aeruginosa PCL-degrading clinical isolate we identified in a screening. We reveal that the mechanism underpinning this enhanced biofilm formation is the incorporation of the plastic breakdown products into the extracellular matrix, leading to enhanced biofilm levels. The level of PCL degradation we show by a clinical isolate and its ability to promote a key virulence and persistence determinant such as biofilm formation indicates that the integrity of any PCL containing medical device, such as sutures or implants, and the condition of patients receiving such devices could be severely compromised by pathogens with this capacity. Given the central role of plastic in healthcare, this should be considered in the future of medical interventions and practice and hospital designs implementing this material
Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Project description:Fluoride is the anionic form of fluorine, the 13th most abundant element in Earth's crust, and it is toxic to many organisms above a threshold concentration. Environmental bacteria can withstand relatively high fluoride concentrations, but the only mechanism described so far is the CrcB-dependent efflux. CrcB-mediated export is the primary mechanism of F-tolerance in the model environmental bacterium Pseudomonas putida, yet spontaneous NaF-tolerant mutants arise in the absence of the CrcB transporter, showing that this is not the sole pathway of tolerance. We used whole-genome sequencing, proteomic, and transcriptomic analyses to identify mechanisms that affect fluoride tolerance in Pseudomonas putida.