Initial Antimicrobial Testing of a Novel Reusable Intermittent Urinary Catheter System and Catheter Reprocessing Device.

OBJECTIVE: To evaluate the efficacy of the Aurie System, a preclinical prototype allowing for standardized intermittent catheter (IC) reuse of novel reusable no-touch ICs. Individuals with neurogenic bladder often require single-use ICs to urinate, but urinary tract infection (UTI) is a common cause of morbidity for IC users. Safer no-touch catheters are not easily affordable, and the Aurie System attempts to provide no-touch catheters at a fraction of the price by allowing for standardized and safe IC reuse. METHODS: Standard ICs were inoculated with Escherichia coli and Pseudomonas aeruginosa and incubated for 48 hours to assess microbial burden and biofilm formation (the latter using infrared fluorescence imaging). This procedure was repeated with Aurie ICs, focusing on evaluating catheter microbial burden after inoculation and reprocessing with the prototype washer-disinfector. This was repeated with up to 100 cycles to evaluate repetitive use. RESULTS: Standard ICs showed bacterial attachment and biofilm development peaking at 24 hours of incubation. The Aurie catheters produced a similar outcome but, after reprocessing, microbial burden was reduced below the level of detection. Repeat cycles showed pathogen clearance to similar levels. One catheter reached 100 cycles and there was no viable pathogen load after reprocessing. CONCLUSION: Intermittent urinary catheters, when cleaned inappropriately, can harbor viable bacteria and biofilm. The Aurie System, when used to disinfect novel reusable ICs within a prototype reprocessing device, can reduce microbial burden below level of detection even after 100 cycles. This suggests the Aurie System may be a feasible technology for safe IC reuse.

Development and applications of a CRISPR activation system for facile genetic overexpression in Candida albicans.

For the fungal pathogen Candida albicans, genetic overexpression readily occurs via a diversity of genomic alterations, such as aneuploidy and gain-of-function mutations, with important consequences for host adaptation, virulence, and evolution of antifungal drug resistance. Given the important role of overexpression on C. albicans biology, it is critical to develop and harness tools that enable the analysis of genes expressed at high levels in the fungal cell. Here, we describe the development, optimization, and application of a novel, single-plasmid-based CRISPR activation (CRISPRa) platform for targeted genetic overexpression in C. albicans, which employs a guide RNA to target an activator complex to the promoter region of a gene of interest, thus driving transcriptional expression of that gene. Using this system, we demonstrate the ability of CRISPRa to drive high levels of gene expression in C. albicans, and we assess optimal guide RNA targeting for robust and constitutive overexpression. We further demonstrate the specificity of the system via RNA sequencing. We highlight the application of CRISPR activation to overexpress genes involved in pathogenesis and drug susceptibility, and contribute toward the identification of novel phenotypes. Consequently, this tool will facilitate a broad range of applications for the study of C. albicans genetic overexpression.