{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T00:01:07Z","timestamp":1793318467518,"version":"4.5.0"},"reference-count":55,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T00:00:00Z","timestamp":1630454400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004359","name":"Vetenskapsr\u00e5det","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007464","name":"Torsten S\u00f6derbergs Stiftelse","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007464","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005390","name":"Alfred \u00d6sterlunds Stiftelse","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005390","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009788","name":"Sten A Olssons Stiftelse f\u00f6r Forskning och Kultur","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009788","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010823","name":"Tore Nilsons Stiftelse f\u00f6r Medicinsk Forskning","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005753","name":"Kungliga Fysiografiska S\u00e4llskapet i Lund","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005753","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008444","name":"O. E. och Edla Johanssons Vetenskapliga Stiftelse","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008444","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004722","name":"Stiftelsen Lars Hiertas Minne","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004722","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008584","name":"Stiftelsen L\u00e4ngmanska Kulturfonden","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008584","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006350","name":"Stiftelsen Clas Groschinskys Minnesfond","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006350","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["AI32296 , AI147930"],"award-info":[{"award-number":["AI32296 , AI147930"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>\n                    <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                    is the etiological agent of gonorrhea, the second most common bacterial sexually transmitted infection worldwide. Reproductive sequelae of gonorrhea include infertility, ectopic pregnancy and chronic pelvic pain. Most antibiotics currently in clinical use have been rendered ineffective due to the rapid spread of antimicrobial resistance among gonococci. The developmental pipeline of new antibiotics is sparse and novel therapeutic approaches are urgently needed. Previously, we utilized the ability of\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    to bind the complement inhibitor C4b-binding protein (C4BP) to evade killing by human complement to design a chimeric protein that linked the two N-terminal gonococcal binding domains of C4BP with the Fc domain of IgM. The resulting molecule, C4BP-IgM, enhanced complement-mediated killing of gonococci. Here we show that C4BP-IgM induced membrane perturbation through complement deposition and membrane attack complex pore insertion facilitates the access of antibiotics to their intracellular targets. Consequently, bacteria become more susceptible to killing by antibiotics. Remarkably, C4BP-IgM restored susceptibility to azithromycin of two azithromycin-resistant clinical gonococcal strains because of overexpression of the MtrC-MtrD-MtrE efflux pump. Our data show that complement activation can potentiate activity of antibiotics and suggest a role for C4BP-IgM as an adjuvant for antibiotic treatment of drug-resistant gonorrhea.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2021.726801","type":"journal-article","created":{"date-parts":[[2021,9,3]],"date-time":"2021-09-03T20:29:47Z","timestamp":1630700987000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":14,"title":["Serum Complement Activation by C4BP-IgM Fusion Protein Can Restore Susceptibility to Antibiotics in Neisseria gonorrhoeae"],"prefix":"10.3389","volume":"12","author":[{"given":"Serena","family":"Bettoni","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karolina","family":"Maziarz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M Rhia L","family":"Stone","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark A T","family":"Blaskovich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Potempa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Luiza","family":"Bazzo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magnus","family":"Unemo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanjay","family":"Ram","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna M.","family":"Blom","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,9,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2017.169","article-title":"Neisseria Gonorrhoeae Host Adaptation and Pathogenesis","volume":"16","author":"Quillin","year":"2018","journal-title":"Nat Rev Microbiol"},{"key":"B2","doi-asserted-by":"publisher","first-page":"548","DOI":"10.2471\/BLT.18.228486","article-title":"Chlamydia, Gonorrhoea, Trichomoniasis and Syphilis: Global Prevalence and Incidence Estimates, 2016","volume":"97","author":"Rowley","year":"2019","journal-title":"Bull World Health Organ"},{"key":"B3","doi-asserted-by":"publisher","first-page":"e2","DOI":"10.1056\/NEJMicm1808613","article-title":"Ophthalmia Neonatorum","volume":"380","author":"Tan","year":"2019","journal-title":"N Engl J Med"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciz145","article-title":"Risk of Ectopic Pregnancy and Tubal Infertility Following Gonorrhea and Chlamydia Infections","volume":"69","author":"Reekie","year":"2019","journal-title":"Clin Infect Dis"},{"key":"B5","article-title":"Pelvic Inflammatory Disease","volume-title":"StatPearls","author":"Jennings","year":"2020"},{"key":"B6","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1136\/sti.75.1.3","article-title":"From Epidemiological Synergy to Public Health Policy and Practice: The Contribution of Other Sexually Transmitted Diseases to Sexual Transmission of HIV Infection","volume":"75","author":"Fleming","year":"1999","journal-title":"Sex Transm Infect"},{"key":"B7","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1038\/s41572-019-0128-6","article-title":"Gonorrhoea","volume":"5","author":"Unemo","year":"2019","journal-title":"Nat Rev Dis Primers"},{"key":"B8","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1177\/0956462412472837","article-title":"2012 European Guideline on the Diagnosis and Treatment of Gonorrhoea in Adults","volume":"24","author":"Bignell","year":"2013","journal-title":"Int J STD AIDS"},{"key":"B9","unstructured":"WHO Guidelines for the Treatment of Neisseria Gonorrhoeae2016"},{"key":"B10","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1177\/0956462419886775","article-title":"2018 UK National Guideline for the Management of Infection With Neisseria Gonorrhoeae","volume":"31","author":"Fifer","year":"2020","journal-title":"Int J STD AIDS"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.15585\/mmwr.mm6950a6","article-title":"Update to CDC's Treatment Guidelines for Gonococcal Infection, 2020","volume":"69","author":"St Cyr","year":"2020","journal-title":"MMWR Morb Mortal Wkly Rep"},{"key":"B12","doi-asserted-by":"publisher","first-page":"956462420949126","DOI":"10.1177\/0956462420949126","article-title":"2020 European Guideline for the Diagnosis and Treatment of Gonorrhoea in Adults","author":"Unemo","year":"2020","journal-title":"Int J STD AIDS"},{"key":"B13","doi-asserted-by":"publisher","first-page":"e1002344","DOI":"10.1371\/journal.pmed.1002344","article-title":"Antimicrobial Resistance in Neisseria Gonorrhoeae: Global Surveillance and a Call for International Collaborative Action","volume":"14","author":"Wi","year":"2017","journal-title":"PloS Med"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1071\/SH19023","article-title":"World Health Organization Global Gonococcal Antimicrobial Surveillance Program (WHO GASP): Review of New Data and Evidence to Inform International Collaborative Actions and Research Efforts","volume":"16","author":"Unemo","year":"2019","journal-title":"Sex Health"},{"key":"B15","volume-title":"Division of Healthcare Quality Promotion. Antibiotic Resistance Coordination and Strategy Unit. Antibiotic resistance threats in the United States, 2019","year":"2019"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(17)30753-3","article-title":"Discovery, Research, and Development of New Antibiotics: The WHO Priority List of Antibiotic-Resistant Bacteria and Tuberculosis","volume":"18","author":"Tacconelli","year":"2018","journal-title":"Lancet Infect Dis"},{"key":"B17","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1128\/CMR.00010-14","article-title":"Antimicrobial Resistance in Neisseria Gonorrhoeae in the 21st Century: Past, Evolution, and Future","volume":"27","author":"Unemo","year":"2014","journal-title":"Clin Microbiol Rev"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-143-7-2117","article-title":"The MtrD Protein of Neisseria Gonorrhoeae Is a Member of the Resistance\/Nodulation\/Division Protein Family Constituting Part of an Efflux System","volume":"143","author":"Hagman","year":"1997","journal-title":"Microbiol (Read)"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkaa040","article-title":"Genomic Analysis and Antimicrobial Resistance of Neisseria Gonorrhoeae Isolates From Vietnam in 2011 and 2015-16","volume":"75","author":"Lan","year":"2020","journal-title":"J Antimicrob Chemother"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01509-20","article-title":"Efflux Pump Antibiotic Binding Site Mutations Are Associated With Azithromycin Nonsusceptibility in Clinical Neisseria Gonorrhoeae Isolates","volume":"11","author":"Ma","year":"2020","journal-title":"mBio"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkaa318","article-title":"Genomic Epidemiology of Neisseria Gonorrhoeae Elucidating the Gonococcal Antimicrobial Resistance and Lineages\/Sublineages Across Brazil, 2015-16","volume":"75","author":"Golparian","year":"2020","journal-title":"J Antimicrob Chemother"},{"key":"B22","doi-asserted-by":"publisher","first-page":"e131886","DOI":"10.1172\/jci.insight.131886","article-title":"C4BP-IgM Protein as a Therapeutic Approach to Treat Neisseria Gonorrhoeae Infections","volume":"4","author":"Bettoni","year":"2019","journal-title":"JCI Insight"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.13.5.820","article-title":"Potentiation of Antibiotic Bactericidal Activity by Normal Human Serum","volume":"13","author":"Dutcher","year":"1978","journal-title":"Antimicrob Agents Chemother"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1159\/000238399","article-title":"Influence of Human Serum on Bactericidal Activity of Ceftizoxime Sodium","volume":"32","author":"Watanabe","year":"1986","journal-title":"Chemotherapy"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/162.4.914","article-title":"Influence of Subinhibitory Concentrations of Cephalosporins on the Serum Sensitivity of Pseudomonas Aeruginosa","volume":"162","author":"Darveau","year":"1990","journal-title":"J Infect Dis"},{"key":"B26","article-title":"Synergistic Effect of Cefotaxime and Normal Human Serum on Bactericidal Activity Against Urinary Strains of Escherichia Coli With Capsular Antigen K1","volume":"42","author":"Franiczek","year":"1993","journal-title":"Acta Microbiol Pol"},{"key":"B27","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.resmic.2015.10.002","article-title":"Improved In Vitro Evaluation of Novel Antimicrobials: Potential Synergy Between Human Plasma and Antibacterial Peptidomimetics, AMPs and Antibiotics Against Human Pathogenic Bacteria","volume":"167","author":"Citterio","year":"2016","journal-title":"Res Microbiol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1128\/iai.6.5.709-717.1972","article-title":"Interaction of Complement and Polymyxin With Gram-Negative Bacteria","volume":"6","author":"Pruul","year":"1972","journal-title":"Infect Immun"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/303526a0","article-title":"Sensitization of Gram-Negative Bacteria to Antibiotics and Complement by a Nontoxic Oligopeptide","volume":"303","author":"Vaara","year":"1983","journal-title":"Nature"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1007\/BF01640908","article-title":"In Vitro and In Vivo Effect of Immunoglobulin G on the Integrity of Bacterial Membranes","volume":"12","author":"Dalhoff","year":"1984","journal-title":"Infection"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.36.1.10","article-title":"Potentiation of Antibacterial Activity of Azithromycin and Other Macrolides by Normal Human Serum","volume":"36","author":"Pruul","year":"1992","journal-title":"Antimicrob Agents Chemother"},{"key":"B32","doi-asserted-by":"publisher","first-page":"3074","DOI":"10.1038\/s41598-019-38577-9","article-title":"Complement-Dependent Outer Membrane Perturbation Sensitizes Gram-Negative Bacteria to Gram-Positive Specific Antibiotics","volume":"9","author":"Heesterbeek","year":"2019","journal-title":"Sci Rep"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1084\/jem.162.6.2017","article-title":"Analyses of Gonococcal H8 Antigen. Surface Location, Inter- and Intrastrain Electrophoretic Heterogeneity, and Unusual Two-Dimensional Electrophoretic Characteristics","volume":"162","author":"Hitchcock","year":"1985","journal-title":"J Exp Med"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(94)36013-8","article-title":"Complement-Mediated Bacterial Killing Assays","volume":"236","author":"McQuillen","year":"1994","journal-title":"Methods Enzymol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-9496-0_16","article-title":"Complement-Dependent Serum Bactericidal Assays for Neisseria Gonorrhoeae","volume":"1997","author":"Gulati","year":"2019","journal-title":"Methods Mol Biol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12388","article-title":"A Novel Method for Direct Measurement of Complement Convertases Activity in Human Serum","volume":"178","author":"Blom","year":"2014","journal-title":"Clin Exp Immunol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1039\/C9MD00124G","article-title":"Fluoroquinolone-Derived Fluorescent Probes for Studies of Bacterial Penetration and Efflux","volume":"10","author":"Stone","year":"2019","journal-title":"Medchemcomm"},{"key":"B38","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1039\/D0CB00118J","article-title":"ORCID Logo, Fluorescent Macrolide Probes \u2013 Synthesis and Use in Evaluation of Bacterial Resistance","volume":"1","author":"Stone","year":"2020","journal-title":"RSC Chem Biol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1111\/eci.12419","article-title":"Compstatin: A C3-Targeted Complement Inhibitor Reaching its Prime for Bedside Intervention","volume":"45","author":"Mastellos","year":"2015","journal-title":"Eur J Clin Invest"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1084\/jem.193.3.281","article-title":"Binding of C4b-Binding Protein to Porin: A Molecular Mechanism of Serum Resistance of Neisseria Gonorrhoeae","volume":"193","author":"Ram","year":"2001","journal-title":"J Exp Med"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2015.11.003","article-title":"The Human Cathelicidin LL-37\u2013A Pore-Forming Antibacterial Peptide and Host-Cell Modulator","volume":"1858","author":"Xhindoli","year":"2016","journal-title":"Biochim Biophys Acta"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0406678101","article-title":"Activation of the Complement System Generates Antibacterial Peptides","volume":"101","author":"Nordahl","year":"2004","journal-title":"Proc Natl Acad Sci USA"},{"key":"B43","first-page":"29","article-title":"Gonococcal Vaccine Studies in Inuvik","volume":"65","author":"Greenberg","year":"1974","journal-title":"Can J Public Health"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/0264-410X(91)90147-X","article-title":"Efficacy Trial of a Parenteral Gonococcal Pilus Vaccine in Men","volume":"9","author":"Boslego","year":"1991","journal-title":"Vaccine"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2011.04.054","article-title":"Strategies for Bypassing the Membrane Barrier in Multidrug Resistant Gram-Negative Bacteria","volume":"585","author":"Bolla","year":"2011","journal-title":"FEBS Lett"},{"key":"B46","doi-asserted-by":"publisher","first-page":"17028","DOI":"10.1038\/nmicrobiol.2017.28","article-title":"Pentamidine Sensitizes Gram-Negative Pathogens to Antibiotics and Overcomes Acquired Colistin Resistance","volume":"2","author":"Stokes","year":"2017","journal-title":"Nat Microbiol"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01193-19","article-title":"HAMLET, a Protein Complex From Human Milk has Bactericidal Activity and Enhances the Activity of Antibiotics Against Pathogenic Streptococci","volume":"63","author":"Alamiri","year":"2019","journal-title":"Antimicrob Agents Chemother"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.2174\/0929867323666160304150522","article-title":"Current Advances in Developing Inhibitors of Bacterial Multidrug Efflux Pumps","volume":"23","author":"Mahmood","year":"2016","journal-title":"Curr Med Chem"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1701666","article-title":"Human Factor H Domains 6 and 7 Fused to IgG1 Fc Are Immunotherapeutic Against Neisseria Gonorrhoeae","volume":"201","author":"Shaughnessy","year":"2018","journal-title":"J Immunol"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.583305","article-title":"Development of Complement Factor H-Based Immunotherapeutic Molecules in Tobacco Plants Against Multidrug-Resistant Neisseria Gonorrhoeae","volume":"11","author":"Shaughnessy","year":"2020","journal-title":"Front Immunol"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2015.01.020","article-title":"Contribution of the Complement Membrane Attack Complex to the Bactericidal Activity of Human Serum","volume":"65","author":"Berends","year":"2015","journal-title":"Mol Immunol"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2020.02.006","article-title":"Complement Membrane Attack Complex: New Roles, Mechanisms of Action, and Therapeutic Targets","volume":"190","author":"Xie","year":"2020","journal-title":"Am J Pathol"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01517.x","article-title":"Induction of the mtrCDE-Encoded Efflux Pump System of Neisseria Gonorrhoeae Requires MtrA, an AraC-Like Protein","volume":"33","author":"Rouquette","year":"1999","journal-title":"Mol Microbiol"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.71.10.5576-5582.2003","article-title":"A Gonococcal Efflux Pump System Enhances Bacterial Survival in a Female Mouse Model of Genital Tract Infection","volume":"71","author":"Jerse","year":"2003","journal-title":"Infect Immun"},{"key":"B55","doi-asserted-by":"publisher","first-page":"4126","DOI":"10.1038\/s41467-020-17980-1","article-title":"Adaptation to the Cervical Environment Is Associated With Increased Antibiotic Susceptibility in Neisseria Gonorrhoeae","volume":"11","author":"Ma","year":"2020","journal-title":"Nat Commun"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.726801\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,3]],"date-time":"2021-09-03T20:29:59Z","timestamp":1630700999000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.726801\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,1]]},"references-count":55,"alternative-id":["10.3389\/fimmu.2021.726801"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2021.726801","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.740841722.793591014","asserted-by":"object"}]},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,1]]},"article-number":"726801"}}