{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T00:48:03Z","timestamp":1759798083526,"version":"build-2065373602"},"reference-count":53,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T00:00:00Z","timestamp":1741651200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005856","name":"Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa","doi-asserted-by":"publisher","award":["UI\/BD\/154472\/2022"],"award-info":[{"award-number":["UI\/BD\/154472\/2022"]}],"id":[{"id":"10.13039\/501100005856","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["advanced.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Adv Healthcare Materials"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Efflux\u2010mediated antibiotic resistance poses a significant global threat, affecting diverse bacterial species. Clinicians recognize the danger of efflux mechanisms during antibiotic treatment, yet precise diagnostic tools remain unavailable. The antibiogram currently infers abnormal efflux pump activity in clinical isolates, which is subsequently confirmed through transcriptomic or genomic analysis. This study harnesses the colorimetric, fluorescence, and solubility properties of phenazinium derivatives to develop a rapid protocol for detecting bacterial efflux. Among several synthesized phenazinium compounds, the compound demonstrating differential MIC in\u00a0<jats:italic>Staphylococcus<\/jats:italic>\u00a0efflux mutants and exhibiting appropriate physicochemical properties is selected. A diagnostic protocol is developed using the selected compound to assess efflux levels, categorized as no, weak, or strong, through colorimetry and spectroscopy techniques. Testing on Gram\u2010positive efflux mutants and clinical <jats:italic>Staphylococcus<\/jats:italic> isolates further validates the approach. In\u2010silico docking analysis confirms the interaction between the chosen compound and the NorA efflux pump in\u00a0<jats:italic>S. aureus<\/jats:italic>. Flow cytometry is employed to re\u2010evaluate the detection assays. The developed molecule and protocol represent the first known method to evaluate efflux levels in any Gram\u2010positive species through a streamlined and user\u2010friendly process. This pioneering test significantly advances the epidemiological analysis of efflux mechanisms and enables more informed therapeutic decision\u2010making, leading to more effective treatment.<\/jats:p>","DOI":"10.1002\/adhm.202404145","type":"journal-article","created":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T23:44:18Z","timestamp":1741736658000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Unlocking the Gates: A Novel Diagnostic Molecule for Quantifying Efflux Levels in Gram\u2010Positive Bacteria"],"prefix":"10.1002","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5722-6567","authenticated-orcid":false,"given":"Mrunal","family":"Patil","sequence":"first","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"},{"name":"Aix Marseille Universit\u00e9 INSERM SSA MCT  Marseille 13385 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3682-6838","authenticated-orcid":false,"given":"Tatiana","family":"Munteanu","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5490-270X","authenticated-orcid":false,"given":"Ga\u00ebl","family":"Brasseur","sequence":"additional","affiliation":[{"name":"Laboratoire de Chimie Bact\u00e9rienne Institut de Microbiologie de la M\u00e9diterran\u00e9e CNRS Aix\u2010Marseille Universit\u00e9  Marseille 13402 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3070-7421","authenticated-orcid":false,"given":"Carolina","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine GHTM Associate Laboratory in Translation and Innovation Towards Global Health LA\u2010REAL Instituto de Higiene e Medicina Tropical IHMT Universidade NOVA de Lisboa UNL  Lisbon 1349\u2010008 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4096-2410","authenticated-orcid":false,"given":"Sofia Santos","family":"Costa","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine GHTM Associate Laboratory in Translation and Innovation Towards Global Health LA\u2010REAL Instituto de Higiene e Medicina Tropical IHMT Universidade NOVA de Lisboa UNL  Lisbon 1349\u2010008 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3536-2733","authenticated-orcid":false,"given":"Isabel","family":"Couto","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine GHTM Associate Laboratory in Translation and Innovation Towards Global Health LA\u2010REAL Instituto de Higiene e Medicina Tropical IHMT Universidade NOVA de Lisboa UNL  Lisbon 1349\u2010008 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6337-1026","authenticated-orcid":false,"given":"Mohd","family":"Athar","sequence":"additional","affiliation":[{"name":"Physics Department University of Cagliari  SP 8, km 0.700 Monserrato 09042 Cagliari Italy"}]},{"given":"Elisa","family":"Asunis","sequence":"additional","affiliation":[{"name":"Physics Department University of Cagliari  SP 8, km 0.700 Monserrato 09042 Cagliari Italy"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4013-8867","authenticated-orcid":false,"given":"Attilio Vittorio","family":"Vargiu","sequence":"additional","affiliation":[{"name":"Physics Department University of Cagliari  SP 8, km 0.700 Monserrato 09042 Cagliari Italy"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9676-6251","authenticated-orcid":false,"given":"Miguel","family":"Viveiros","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine GHTM Associate Laboratory in Translation and Innovation Towards Global Health LA\u2010REAL Instituto de Higiene e Medicina Tropical IHMT Universidade NOVA de Lisboa UNL  Lisbon 1349\u2010008 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4962-3191","authenticated-orcid":false,"given":"Carole","family":"DiGiorgio","sequence":"additional","affiliation":[{"name":"Laboratoire de Mutag\u00e9n\u00e8se Environnementale Aix\u2010Marseille Universit\u00e9 CNRS IRD Avignon Universit\u00e9  IMBE UMR 7263 Marseille 13385 France"}]},{"given":"Fr\u00e9d\u00e9ric","family":"Brunel","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4090-0479","authenticated-orcid":false,"given":"Jean\u2010Manuel","family":"Raimundo","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4446-0661","authenticated-orcid":false,"given":"Michel","family":"Camplo","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9747-3813","authenticated-orcid":false,"given":"Olivier","family":"Siri","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 CNRS CINaM UMR 7325 Campus de Luminy  Case 913, Cedex 09 Marseille 13288 France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3892-4340","authenticated-orcid":false,"given":"Jean\u2010Michel","family":"Bolla","sequence":"additional","affiliation":[{"name":"Aix Marseille Universit\u00e9 INSERM SSA MCT  Marseille 13385 France"}]}],"member":"311","published-online":{"date-parts":[[2025,3,11]]},"reference":[{"key":"e_1_2_8_1_1","volume":"4","author":"Bassetti M.","year":"2022","journal-title":"JAC\u2010AMR"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1136\/bmjgh-2021-008159"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10020209"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00066-16"},{"key":"e_1_2_8_5_1","unstructured":"Health Action International Lack of diagnostic use hampering fight against antimicrobial resistance https:\/\/haiweb.org\/media\u2010resource\/lack\u2010of\u2010diagnostic\u2010use\u2010hampering\u2010fight\u2010against\u2010antimicrobial\u2010resistance\/2023 (accessed: February 2024)."},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-65981-3"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.drudis.2020.09.028"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.001333"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics13203246"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00840-16"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00635a022"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhazmat.2021.127171"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2018.036"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.3390\/ani13071270"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics9100639"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.1c05933"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42004-020-0326-x"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2021.104894"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01058-10"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-11-241"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.2174\/1874285801307010072"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/ardp.202300103"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.12.2650"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.47.2.719-726.2003"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi0362018"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics11121837"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi300762p"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics12071168"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10863-021-09886-4"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.6b01439"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm401808n"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2006.09.001"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.3934\/medsci.2022018"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.5.1857-1864.2005"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2020.104452"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00672-10"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/42.6.807"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2017.09.001"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.43.10.2404"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-022-00994-9"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms20164041"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.2174\/1874285801307010059"},{"volume-title":"Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically","year":"2012","author":"Clinical and Laboratory Standards Institute (CLSI)","key":"e_1_2_8_43_1"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.2147\/IDR.S193649"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.9b03698"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-20558-z"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02762"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.1c00203"},{"key":"e_1_2_8_49_1","unstructured":"M. J.Frisch G. W.Trucks H. B.Schlegel G. E.Scuseria M. A.Robb J. R.Cheeseman G.Scalmani V.Barone G. A.Petersson H.Nakatsuji X.Li M.Caricato A. V.Marenich J.Bloino B. G.Janesko R.Gomperts B.Mennucci H. P.Hratchian J. V.Ortiz A. F.Izmaylov J. L.Sonnenberg D.Williams\u2010Young F.Ding F.Lipparini F.Egidi J.Goings B.Peng A.Petrone T.Henderson D.Ranasinghe et\u00a0al.Gaussian 16 Rev. C. 01 Gaussian Inc. Wallingford CT (2016) GaussView 5.0. Wallingford E.U.A 2016."},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules200813997"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41597-022-01261-1"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkn217"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkx400"}],"container-title":["Advanced Healthcare Materials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/advanced.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/adhm.202404145","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T15:40:19Z","timestamp":1759765219000},"score":1,"resource":{"primary":{"URL":"https:\/\/advanced.onlinelibrary.wiley.com\/doi\/10.1002\/adhm.202404145"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,11]]},"references-count":53,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/adhm.202404145"],"URL":"https:\/\/doi.org\/10.1002\/adhm.202404145","archive":["Portico"],"relation":{},"ISSN":["2192-2640","2192-2659"],"issn-type":[{"type":"print","value":"2192-2640"},{"type":"electronic","value":"2192-2659"}],"subject":[],"published":{"date-parts":[[2025,3,11]]},"assertion":[{"value":"2024-10-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-03-11","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"2404145"}}