{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T14:44:36Z","timestamp":1790779476734,"version":"4.1.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2017,3,1]],"date-time":"2017-03-01T00:00:00Z","timestamp":1488326400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100003554","name":"Lundbeckfonden","doi-asserted-by":"publisher","award":["R105-A9791"],"award-info":[{"award-number":["R105-A9791"]}],"id":[{"id":"10.13039\/501100003554","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2017,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Biofilm infections caused by\n            <jats:named-content content-type=\"genus-species\">Pseudomonas aeruginosa<\/jats:named-content>\n            are frequently treated with ciprofloxacin (CIP); however, resistance rapidly develops. One of the primary resistance mechanisms is the overexpression of the MexCD-OprJ pump due to a mutation in\n            <jats:italic>nfxB<\/jats:italic>\n            , encoding the transcriptional repressor of this pump. The aim of this study was to investigate the effect of subinhibitory concentrations of CIP on the occurrence of\n            <jats:italic>nfxB<\/jats:italic>\n            mutants in the wild-type PAO1 flow cell biofilm model. For this purpose, we constructed fluorescent reporter strains (PAO1 background) with an\n            <jats:italic>mCherry<\/jats:italic>\n            tag for constitutive red fluorescence and chromosomal transcriptional fusion between the P\n            <jats:sub>\n              <jats:italic>mexCD<\/jats:italic>\n            <\/jats:sub>\n            promoter and\n            <jats:italic>gfp<\/jats:italic>\n            leading to green fluorescence upon mutation of\n            <jats:italic>nfxB<\/jats:italic>\n            . We observed a rapid development of\n            <jats:italic>nfxB<\/jats:italic>\n            mutants by live confocal laser scanning microscopy (CLSM) imaging of the flow cell biofilm (reaching 80 to 90% of the whole population) when treated with 1\/10 minimal biofilm inhibitory concentration of CIP for 24 h and 96 h. Based on the observed developmental stages, we propose that\n            <jats:italic>nfxB<\/jats:italic>\n            mutants emerged\n            <jats:italic>de novo<\/jats:italic>\n            in the biofilm during CIP treatment from filamentous cells, which might have arisen due to the stress responses induced by CIP. Identical\n            <jats:italic>nfxB<\/jats:italic>\n            mutations were found in fluorescent colonies from the same flow cell biofilm, especially in 24-h biofilms, suggesting selection and clonal expansion of the mutants during biofilm growth. Our findings point at the significant role of high-enough antibiotic dosages or appropriate combination therapy to avoid the emergence of resistant mutants in biofilms.\n          <\/jats:p>","DOI":"10.1128\/aac.02292-16","type":"journal-article","created":{"date-parts":[[2016,12,23]],"date-time":"2016-12-23T17:28:40Z","timestamp":1482514120000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":42,"title":["Real-Time Monitoring of\n            <i>nfxB<\/i>\n            Mutant Occurrence and Dynamics in Pseudomonas aeruginosa Biofilm Exposed to Subinhibitory Concentrations of Ciprofloxacin"],"prefix":"10.1128","volume":"61","author":[{"given":"Greta","family":"Zaborskyte","sequence":"first","affiliation":[{"name":"Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens 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