{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T07:15:46Z","timestamp":1789888546576,"version":"4.0.1"},"reference-count":46,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2017,2,1]],"date-time":"2017-02-01T00:00:00Z","timestamp":1485907200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001186","name":"Australian Cystic Fibrosis Research Trust","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001186","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":[[2017,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            PYRRO-C3D is a cephalosporin-3-diazeniumdiolate nitric oxide (NO) donor prodrug designed to selectively deliver NO to bacterial infection sites. The objective of this study was to assess the activity of PYRRO-C3D against nontypeable\n            <jats:named-content content-type=\"genus-species\">Haemophilus influenzae<\/jats:named-content>\n            (NTHi) biofilms and examine the role of NO in reducing biofilm-associated antibiotic tolerance. The activity of PYRRO-C3D on\n            <jats:italic>in vitro<\/jats:italic>\n            NTHi biofilms was assessed through CFU enumeration and confocal microscopy. NO release measurements were performed using an ISO-NO probe. NTHi biofilms grown on primary ciliated respiratory epithelia at an air-liquid interface were used to investigate the effects of PYRRO-C3D in the presence of host tissue. Label-free liquid chromatography-mass spectrometry (LC\/MS) proteomic analyses were performed to identify differentially expressed proteins following NO treatment. PYRRO-C3D specifically released NO in the presence of NTHi, while no evidence of spontaneous NO release was observed when the compound was exposed to primary epithelial cells. NTHi lacking \u03b2-lactamase activity failed to trigger NO release. Treatment significantly increased the susceptibility of\n            <jats:italic>in vitro<\/jats:italic>\n            NTHi biofilms to azithromycin, causing a log fold reduction (10-fold reduction or 1-log-unit reduction) in viability (\n            <jats:italic>P<\/jats:italic>\n            &lt; 0.05) relative to azithromycin alone. The response was more pronounced for biofilms grown on primary respiratory epithelia, where a 2-log-unit reduction was observed (\n            <jats:italic>P<\/jats:italic>\n            &lt; 0.01). Label-free proteomics showed that NO increased expression of 16 proteins involved in metabolic and transcriptional\/translational functions. NO release from PYRRO-C3D enhances the efficacy of azithromycin against NTHi biofilms, putatively via modulation of NTHi metabolic activity. Adjunctive therapy with NO mediated through PYRRO-C3D represents a promising approach for reducing biofilm-associated antibiotic tolerance.\n          <\/jats:p>","DOI":"10.1128\/aac.02086-16","type":"journal-article","created":{"date-parts":[[2016,12,6]],"date-time":"2016-12-06T03:00:36Z","timestamp":1480993236000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":32,"title":["Cephalosporin-3\u2032-Diazeniumdiolate NO Donor Prodrug PYRRO-C3D Enhances Azithromycin Susceptibility of Nontypeable Haemophilus influenzae Biofilms"],"prefix":"10.1128","volume":"61","author":[{"given":"Samuel A.","family":"Collins","sequence":"first","affiliation":[{"name":"Clinical and Experimental Sciences, Faculty of Medicine and Institute for Life Sciences, University of Southampton, Southampton, United Kingdom"},{"name":"Southampton NIHR Respiratory Biomedical Research Unit, University Hospital Southampton NHS Foundation Trust, 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