{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T11:53:04Z","timestamp":1790077984092,"version":"4.0.1"},"reference-count":60,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,11,15]],"date-time":"2018-11-15T00:00:00Z","timestamp":1542240000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Federal State of Lower Saxony","award":["Nieders\u00e4chsisches Vorab (VWZN2889)"],"award-info":[{"award-number":["Nieders\u00e4chsisches Vorab (VWZN2889)"]}]},{"name":"Federal State of Mecklenburg-Pomerania","award":["UG 14 001"],"award-info":[{"award-number":["UG 14 001"]}]},{"name":"Federal State of Mecklenburg-Pomerania","award":["UG 16 001"],"award-info":[{"award-number":["UG 16 001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The anaerobic pathogen Clostridium difficile is of growing significance for the health care system due to its increasing incidence and mortality. As C. difficile infection is both supported and treated by antibiotics, a deeper knowledge on how antimicrobial agents affect the physiology of this important pathogen may help to understand and prevent the development and spreading of antibiotic resistant strains. As the proteomic response of a cell to stress aims at counteracting the harmful effects of this stress, it can be expected that the pattern of a pathogen\u2019s responses to antibiotic treatment will be dependent on the antibiotic mechanism of action. Hence, every antibiotic treatment is expected to result in a specific proteomic signature characterizing its mode of action. In the study presented here, the proteomic response of C. difficile 630\u2206erm to vancomycin, metronidazole, and fidaxomicin stress was investigated on the level of protein abundance and protein synthesis based on 2D PAGE. The quantification of 425 proteins of C. difficile allowed the deduction of proteomic signatures specific for each drug treatment. Indeed, these proteomic signatures indicate very specific cellular responses to each antibiotic with only little overlap of the responses. Whereas signature proteins for vancomycin stress fulfil various cellular functions, the proteomic signature of metronidazole stress is characterized by alterations of proteins involved in protein biosynthesis and protein degradation as well as in DNA replication, recombination, and repair. In contrast, proteins differentially expressed after fidaxomicin treatment can be assigned to amino acid biosynthesis, transcription, cell motility, and the cell envelope functions. Notably, the data provided by this study hint also at so far unknown antibiotic detoxification mechanisms.<\/jats:p>","DOI":"10.3390\/cells7110213","type":"journal-article","created":{"date-parts":[[2018,11,15]],"date-time":"2018-11-15T11:32:47Z","timestamp":1542281567000},"page":"213","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Proteomic Signatures of Clostridium difficile Stressed with Metronidazole, Vancomycin, or Fidaxomicin"],"prefix":"10.3390","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6573-1088","authenticated-orcid":false,"given":"Sandra","family":"Maa\u00df","sequence":"first","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Proteomics, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Otto","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Proteomics, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dirk","family":"Albrecht","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Physiology and Molecular Biology, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina","family":"Riedel","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Physiology and Molecular Biology, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anke","family":"Trautwein-Schult","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Proteomics, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D\u00f6rte","family":"Becher","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Department of Microbial Proteomics, University of Greifswald, Felix-Hausdorff-Str. 8, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,11,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.anaerobe.2016.06.008","article-title":"Reclassification of Clostridium difficile as Clostridioides difficile (Hall and O\u2019Toole 1935) Pr\u00e9vot 1938","volume":"40","author":"Lawson","year":"2016","journal-title":"Anaerobe"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1007\/s40273-014-0161-y","article-title":"A review of the economics of treating Clostridium difficile infection","volume":"32","author":"Mergenhagen","year":"2014","journal-title":"Pharmacoeconomics"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1093\/jac\/36.4.673","article-title":"Diarrhoea caused by Clostridium difficile: Response time for treatment with metronidazole and vancomycin","volume":"36","author":"Wilcox","year":"1995","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_4","first-page":"127","article-title":"Treatment of Clostridium difficile-associated diarrhea and colitis","volume":"250","author":"Gerding","year":"2000","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1038\/ajg.2013.4","article-title":"Guidelines for diagnosis, treatment, and prevention of Clostridium difficile infections","volume":"108","author":"Surawicz","year":"2013","journal-title":"Am. J. Gastroenterol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1042\/bj1110195","article-title":"Specificity of combination between mucopeptide precursors and vancomycin or ristocetin","volume":"111","author":"Perkins","year":"1969","journal-title":"Biochem. J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"S206","DOI":"10.1086\/516247","article-title":"Is there a relationship between vancomycin-resistant enterococcal infection and Clostridium difficile infection?","volume":"25","author":"Gerding","year":"1997","journal-title":"Clin. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gajd\u00e1cs, M., Spengler, G., and Urb\u00e1n, E. (2017). Identification and antimicrobial susceptibility testing of anaerobic bacteria: Rubik\u2019s cube of clinical microbiology?. Antibiotics, 6.","DOI":"10.3390\/antibiotics6040025"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/j.idc.2016.02.006","article-title":"Vancomycin-resistant enterococci: Therapeutic challenges in the 21st century","volume":"30","author":"Miller","year":"2016","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2607","DOI":"10.1128\/AAC.43.11.2607","article-title":"Antimicrobial susceptibilities and serogroups of clinical strains of Clostridium difficile isolated in France in 1991 and 1997","volume":"43","author":"Barbut","year":"1999","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1647","DOI":"10.1128\/AAC.46.6.1647-1650.2002","article-title":"Reassessment of Clostridium difficile susceptibility to metronidazole and vancomycin","volume":"46","author":"Alonso","year":"2002","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0732-8893(98)00139-4","article-title":"Susceptibility testing of Clostridium difficile against metronidazole and vancomycin by disk diffusion and Etest","volume":"34","author":"Wong","year":"1999","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1093\/jac\/48.5.741","article-title":"Reduced susceptibility of Clostridium difficile to metronidazole","volume":"48","author":"Brazier","year":"2001","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1097\/WNF.0b013e3182334b35","article-title":"Metronidazole-induced central nervous system toxicity: A systematic review","volume":"34","author":"Kuriyama","year":"2011","journal-title":"Clin. Neuropharmacol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.mayocp.2012.07.016","article-title":"Clostridium difficile infection: New insights into management","volume":"87","author":"Khanna","year":"2012","journal-title":"Mayo Clin. Proc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1128\/AAC.01443-07","article-title":"Opt-80 eliminates Clostridium difficile and is sparing of Bacteroides species during treatment of C. difficile infection","volume":"53","author":"Louie","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1128\/AAC.01045-07","article-title":"Safety, tolerance, and pharmacokinetic studies of Opt-80 in healthy volunteers following single and multiple oral doses","volume":"52","author":"Shue","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"S127","DOI":"10.1093\/cid\/cis358","article-title":"Fidaxomicin is an inhibitor of the initiation of bacterial RNA synthesis","volume":"55","author":"Artsimovitch","year":"2012","journal-title":"Clin. Infect. Dis. Off. Publ. Infect. Dis. Soc. Am."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2892","DOI":"10.1093\/jac\/dku261","article-title":"Clinical efficacy of fidaxomicin compared with vancomycin and metronidazole in Clostridium difficile infections: A meta-analysis and indirect treatment comparison","volume":"69","author":"Cornely","year":"2014","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"S132","DOI":"10.1093\/cid\/cis338","article-title":"Fidaxomicin preserves the intestinal microbiome during and after treatment of Clostridium difficile infection (CDI) and reduces both toxin reexpression and recurrence of CDI","volume":"55","author":"Louie","year":"2012","journal-title":"Clin. Infect. Dis. Off. Publ. Infect. Dis. Soc. Am."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/S0168-6445(98)00011-4","article-title":"Eubacterial sigma-factors","volume":"22","year":"1998","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1093\/jac\/47.2.244","article-title":"Antibiotic activity against genotypically distinct and indistinguishable Clostridium difficile isolates","volume":"47","author":"Freeman","year":"2001","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Fuchs, S., Z\u00fchlke, D., Pan\u00e9-Farr\u00e9, J., Kusch, H., Wolf, C., Rei\u00df, S., Binh, L.T.N., Albrecht, D., Riedel, K., and Hecker, M. (2013). Aureolib\u2014A proteome signature library: Towards an understanding of Staphylococcus aureus pathophysiology. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0070669"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4958","DOI":"10.1002\/pmic.200800278","article-title":"Gel-based proteomics of Gram-positive bacteria: A powerful tool to address physiological questions","volume":"8","author":"Hecker","year":"2008","journal-title":"Proteomics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1128\/AAC.47.3.948-955.2003","article-title":"Proteomic approach to understanding antibiotic action","volume":"47","author":"Bandow","year":"2003","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1099\/jmm.0.45790-0","article-title":"Generation of an erythromycin-sensitive derivative of Clostridium difficile strain 630 (630\u0394erm) and demonstration that the conjugative transposon Tn916\u0394E enters the genome of this strain at multiple sites","volume":"54","author":"Hussain","year":"2005","journal-title":"J. Med. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1101\/gr.905003","article-title":"Bacillus subtilis during feast and famine: Visualization of the overall regulation of protein synthesis during glucose starvation by proteome analysis","volume":"13","author":"Bernhardt","year":"2003","journal-title":"Genome Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.mimet.2006.09.018","article-title":"A Standard Operating Procedure (SOP) for the preparation of intra- and extracellular proteins of Clostridium acetobutylicum for proteome analysis","volume":"68","author":"Schwarz","year":"2007","journal-title":"J. Microbiol. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1002\/elps.201200618","article-title":"The new horizon in 2D electrophoresis-new technology to increase resolution and sensitivity","volume":"34","author":"Moche","year":"2013","journal-title":"Electrophoresis"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1099\/jmm.0.000427","article-title":"Manual curation and reannotation of the genomes of Clostridium difficile 630\u0394erm and Clostridium difficile 630","volume":"66","author":"Dannheim","year":"2017","journal-title":"J. Med. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e02383-14","DOI":"10.1128\/mBio.02383-14","article-title":"High-throughput analysis of gene essentiality and sporulation in Clostridium difficile","volume":"6","author":"Dembek","year":"2015","journal-title":"mBio"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Lechner, M., Findei\u00df, S., Steiner, L., Marz, M., Stadler, P.F., and Prohaska, S.J. (2011). Proteinortho: Detection of (Co-)orthologs in large-scale analysis. BMC Bioinform., 12.","DOI":"10.1186\/1471-2105-12-124"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3256","DOI":"10.1002\/pmic.201100046","article-title":"Proteomic signatures in antibiotic research","volume":"11","author":"Wenzel","year":"2011","journal-title":"Proteomics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.anaerobe.2009.12.002","article-title":"Bactericidal activity of telavancin, vancomycin and metronidazole against Clostridium difficile","volume":"16","author":"Goldstein","year":"2010","journal-title":"Anaerobe"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1093\/nar\/gkg128","article-title":"The TIGRFAMs database of protein families","volume":"31","author":"Haft","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/0006-2952(86)90555-1","article-title":"In vitro reductive activation of nitroimidazoles","volume":"35","author":"Declerck","year":"1986","journal-title":"Biochem. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Neumann-Schaal, M., Hofmann, J.D., Will, S.E., and Schomburg, D. (2015). Time-resolved amino acid uptake of Clostridium difficile 630\u0394erm and concomitant fermentation product and toxin formation. BMC Microbiol., 15.","DOI":"10.1186\/s12866-015-0614-2"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2260","DOI":"10.1074\/mcp.M113.035741","article-title":"Highly precise quantification of protein molecules per cell during stress and starvation responses in Bacillus subtilis","volume":"13","author":"Wachlin","year":"2014","journal-title":"Mol. Cell. Proteomics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1016\/j.ijmm.2013.08.014","article-title":"Global proteome analysis of vancomycin stress in Staphylococcus aureus","volume":"303","author":"Hessling","year":"2013","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5589","DOI":"10.1073\/pnas.87.15.5589","article-title":"Ribosomes as sensors of heat and cold shock in Escherichia coli","volume":"87","author":"VanBogelen","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Chong, P.M., Lynch, T., McCorrister, S., Kibsey, P., Miller, M., Gravel, D., Westmacott, G.R., and Mulvey, M.R. (2014). Canadian Nosocomial Infection Surveillance Program (CNISP). Proteomic analysis of a NAP1 Clostridium difficile clinical isolate resistant to metronidazole. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0082622"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1099\/jmm.0.47657-0","article-title":"Microarray analysis of the transcriptional responses of Clostridium difficile to environmental and antibiotic stress","volume":"57","author":"Emerson","year":"2008","journal-title":"J. Med. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1016\/j.resmic.2010.04.003","article-title":"Bacteroides fragilis RecA protein overexpression causes resistance to metronidazole","volume":"161","author":"Steffens","year":"2010","journal-title":"Res. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5082","DOI":"10.1128\/JB.182.18.5082-5090.2000","article-title":"Sequential inactivation of rdxA (HP0954) and frxA(HP0642) nitroreductase genes causes moderate and high-level metronidazole resistance in Helicobacter pylori","volume":"182","author":"Jeong","year":"2000","journal-title":"J. Bacteriol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1093\/jac\/dkh296","article-title":"Metronidazole resistance in Bacteroides spp. carrying nim genes and the selection of slow-growing metronidazole-resistant mutants","volume":"54","author":"Gal","year":"2004","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1111\/j.1574-6968.2000.tb09193.x","article-title":"Frame-shift mutations in NAD(P)H flavin oxidoreductase encoding gene (frxA) from metronidazole resistant Helicobacter pylori ATCC43504 and its involvement in metronidazole resistance","volume":"188","author":"Kwon","year":"2000","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/S0966-842X(99)01454-7","article-title":"Bacterial tellurite resistance","volume":"7","author":"Taylor","year":"1999","journal-title":"Trends Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1111\/j.1574-6976.2009.00177.x","article-title":"Tellurite: History, oxidative stress, and molecular mechanisms of resistance","volume":"33","author":"Chasteen","year":"2009","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"28002","DOI":"10.1038\/srep28002","article-title":"Mycobacterium tuberculosis Rv1152 is a novel GntR family transcriptional regulator involved in intrinsic vancomycin resistance and is a potential vancomycin adjuvant target","volume":"6","author":"Zeng","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Hu, J., Zhao, L., and Yang, M. (2015). A GntR family transcription factor positively regulates mycobacterial isoniazid resistance by controlling the expression of a putative permease. BMC Microbiol., 15.","DOI":"10.1186\/s12866-015-0556-8"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"6122","DOI":"10.1128\/jb.179.19.6122-6126.1997","article-title":"Role of the acrAB locus in organic solvent tolerance mediated by expression of marA, soxS, or robA in Escherichia coli","volume":"179","author":"White","year":"1997","journal-title":"J. Bacteriol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1007\/s007920050066","article-title":"Improvement of organic solvent tolerance level of Escherichia coli by overexpression of stress-responsive genes","volume":"2","author":"Aono","year":"1998","journal-title":"Extremophiles"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1128\/jb.176.1.143-148.1994","article-title":"Repressor mutations in the marRAB operon that activate oxidative stress genes and multiple antibiotic resistance in Escherichia coli","volume":"176","author":"Ariza","year":"1994","journal-title":"J. Bacteriol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2770","DOI":"10.1128\/AAC.41.12.2770","article-title":"Selection of multiple-antibiotic-resistant (mar) mutants of Escherichia coli by using the disinfectant pine oil: Roles of the mar and acrAB loci","volume":"41","author":"Moken","year":"1997","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1111\/j.1574-6968.1998.tb13905.x","article-title":"Overexpression of marA, soxS, or acrAB produces resistance to triclosan in laboratory and clinical strains of Escherichia coli","volume":"166","author":"McMurry","year":"1998","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1128\/jb.178.1.306-308.1996","article-title":"AcrAB efflux pump plays a major role in the antibiotic resistance phenotype of Escherichia coli multiple-antibiotic-resistance (Mar) mutants","volume":"178","author":"Okusu","year":"1996","journal-title":"J. Bacteriol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1128\/AAC.40.5.1266","article-title":"Multiple antibiotic resistance (mar) locus protects Escherichia coli from rapid cell killing by fluoroquinolones","volume":"40","author":"Goldman","year":"1996","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1016\/S0966-842X(99)01589-9","article-title":"The mar regulon: Multiple resistance to antibiotics and other toxic chemicals","volume":"7","author":"Alekshun","year":"1999","journal-title":"Trends Microbiol."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Spengler, G., Kincses, A., Gajd\u00e1cs, M., and Amaral, L. (2017). New roads leading to old destinations: Efflux pumps as targets to reverse multidrug resistance in bacteria. Molecules, 22.","DOI":"10.3390\/molecules22030468"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/7\/11\/213\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:29:57Z","timestamp":1760196597000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/7\/11\/213"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,15]]},"references-count":60,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["cells7110213"],"URL":"https:\/\/doi.org\/10.3390\/cells7110213","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,15]]}}}