Previous work has shown the mprF protein is comprised of two functional domains, the C-terminal and N-terminal. While the C-terminal could independently complete lysinylation of membrane phospholipids, the N-terminal was incapable of completing functions without the assistance of the C-terminal domain. The Q326Stop mutation would logically render the mprF protein non-functional. While our study is novel in examining a large collection of DNS S. aureus strains for stability and PAP, it does have limitations. Firstly, due to the relative rarity of DNS S. aureus, our collection of examined isolates is small at 12 and we were only able to obtain a single daptomycin susceptible—DNS

click here isogenic pair for comparison evaluation. We also used standard inocula (log 106 CFU/mL) for broth microdilution and Etest susceptibility testing per CLSI and manufacturer’s instructions, respectively. The results may have been different if we employed a high inoculum for susceptibility testing (109 CFU/mL)

as was done for the PAP and in vitro PK/PD model of SEVs. Our study is also limited as it focused on the most common gene mutation in DNS S. aureus, mprf, and did not examine the isolates for mutations or changes in expression of other genes known to be involved in DNS S. aureus. Lastly, Protein Tyrosine Kinase inhibitor our isolates are from a single geographic area (Detroit, MI, USA) with an established history of cutting edge resistance in S. aureus and may not be representative of resistance patterns in other areas of the country. Conclusion All 12 DNS S. aureus isolates were stable and displayed different degrees of susceptibility when examined by PAP. To our knowledge, this is the first study to examine such a large collection of clinical DNS S. aureus strains and confirm their stability. This is also the first study to examine the impact of the daptomycin PAP on the activity

of both standard and high dose simulated daptomycin. Additionally, an organism with a unique mutation in mprF, Q326Stop, which would likely render the mprF protein non-functional, was discovered. The findings are clinically relevant because for some organisms the daptomycin AUC predicted antimicrobial activity or killing pattern better than the MIC value by BMD. This highlights the need to consider the Benzatropine whole population of bacteria when discussing susceptibility or the development of resistance. Salubrinal solubility dmso Despite previous reports that some aspects of DNS may be inducible and unstable, eleven of our twelve isolates displayed stable resistance even after 2 years of freezer storage confirming that DNS can frequently be a stable and not transient phenomenon in S. aureus. Daptomycin should continue to be utilized appropriately to minimize resistance and preserve its efficacy. Acknowledgments This study was funded by an investigator initiated grant from Cubist pharmaceuticals. Michael J.