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Mutations in the MexAB-OprM efflux system, naturally occurring in cystic fibrosis (CF) isolates, have been previously shown to reverse this intrinsic resistance. In the present study, we measured temocillin activity in a large collection (n\u2009=\u2009333) of <jats:italic>P. aeruginosa<\/jats:italic> CF isolates. 29% of the isolates had MICs\u2009\u2264\u200916\u2009mg\/L (proposed clinical breakpoint for temocillin). Mutations were observed in <jats:italic>mexA<\/jats:italic> or <jats:italic>mexB<\/jats:italic> in isolates for which temocillin MIC was \u2264512\u2009mg\/L (nucleotide insertions or deletions, premature termination, tandem repeat, nonstop, and missense mutations). A correlation was observed between temocillin MICs and efflux rate of <jats:italic>N<\/jats:italic>-phenyl-1-naphthylamine (MexAB-OprM fluorescent substrate) and extracellular exopolysaccharide abundance (contributing to a mucoid phenotype). OpdK or OpdF anion-specific porins expression decreased temocillin MIC by ~1 two-fold dilution only. Contrarily to the common assumption that temocillin is inactive on <jats:italic>P. aeruginosa<\/jats:italic>, we show here clinically-exploitable MICs on a non-negligible proportion of CF isolates, explained by a wide diversity of mutations in <jats:italic>mexA<\/jats:italic> and\/or <jats:italic>mexB<\/jats:italic>. In a broader context, this work contributes to increase our understanding of MexAB-OprM functionality and help delineating how antibiotics interact with MexA and MexB.<\/jats:p>","DOI":"10.1038\/srep40208","type":"journal-article","created":{"date-parts":[[2017,1,16]],"date-time":"2017-01-16T10:45:47Z","timestamp":1484563547000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":39,"title":["Mechanisms of intrinsic resistance and acquired susceptibility of Pseudomonas aeruginosa isolated from cystic fibrosis patients to temocillin, a revived antibiotic"],"prefix":"10.1038","volume":"7","author":[{"given":"Hussein","family":"Chalhoub","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Pletzer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helge","family":"Weingart","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yvonne","family":"Braun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael M.","family":"Tunney","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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