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Clinical infections are costly and becoming increasingly dangerous as bacteria that once responded to standard antibiotic treatment are developing resistance mechanisms that require innovative treatment strategies. Nitric oxide (NO) is a gaseous molecule produced endogenously that has shown potent antibacterial capabilities in numerous research studies. Its multimechanistic antibacterial methods prevent the development of resistance and have shown potential as an alternative to antibiotics. However, there has yet to be a direct comparison study evaluating the antibacterial properties of NO against antibiotic susceptible and antibiotic\u2010resistant clinically isolated bacterial strains. Herein, standardized lab and clinically isolated drug\u2010resistant bacterial strains are compared side\u2010by\u2010side for growth and viability following treatment with NO released from <jats:italic>S<\/jats:italic>\u2010nitrosoglutathione (GSNO), an NO donor molecule. Evaluation of growth kinetics revealed complete killing of <jats:italic>E. coli<\/jats:italic> lab and clinical strains at 17.5\u2009mM GSNO, though 15\u2009mM displayed &gt;50% killing and significantly reduced metabolic activity, with greater dose dependence for membrane permeability. Clinical <jats:italic>P. aeruginosa<\/jats:italic> showed greater susceptibility to GSNO during growth curve studies, but metabolic activity and membrane permeability demonstrated similar effects for 12.5\u2009mM GSNO treatment of lab and clinical strains. MRSA lab and clinical strains exhibited total killing at 17.5\u2009mM treatment, though metabolic activity was decreased, and membrane permeation began at 12.5\u2009mM for both strains. Lastly, both <jats:italic>S. epidermidis<\/jats:italic> strains were killed by 15\u2009mM GSNO, with sensitivities in metabolic activity and membrane permeability at 12.5\u2009mM GSNO. The mirrored antibacterial effects seen by the lab and clinical strains of two Gram\u2010negative and two Gram\u2010positive bacteria reveal the translational success of NO as an antibacterial therapy and potential alternative to standard antibiotic treatment.<\/jats:p>","DOI":"10.1002\/jbm.b.35442","type":"journal-article","created":{"date-parts":[[2024,6,24]],"date-time":"2024-06-24T12:11:36Z","timestamp":1719231096000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Analysis of the broad\u2010spectrum potential of nitric oxide for antibacterial activity against clinically isolated drug\u2010resistant bacteria"],"prefix":"10.1002","volume":"112","author":[{"given":"Lori M.","family":"Estes Bright","sequence":"first","affiliation":[{"name":"School of Chemical, Materials, and Biomedical Engineering, College of Engineering University of Georgia  Athens Georgia 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