{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T01:03:18Z","timestamp":1785459798080,"version":"3.56.0"},"reference-count":51,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T00:00:00Z","timestamp":1609372800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["2R15AI047115-05A1"],"award-info":[{"award-number":["2R15AI047115-05A1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["SC3GM125556"],"award-info":[{"award-number":["SC3GM125556"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Clinical resistance to amikacin and other aminoglycosides is usually due to the enzymatic acetylation of the antimicrobial molecule. A ubiquitous resistance enzyme among Gram-negatives is the aminoglycoside 6\u2032-N-acetyltransferase type Ib [AAC(6\u2032)-Ib], which catalyzes acetylation using acetyl-CoA as a donor substrate. Therapies that combine the antibiotic and an inhibitor of the inactivation reaction could be an alternative to treat infections caused by resistant bacteria. We previously observed that metal ions such as Zn2+ or Cu2+ in complex with ionophores interfere with the AAC(6\u2032)-Ib-mediated inactivation of aminoglycosides and reduced resistance to susceptibility levels. Ag1+ recently attracted attention as a potentiator of aminoglycosides\u2032 action by mechanisms still in discussion. We found that silver acetate is also a robust inhibitor of the enzymatic acetylation mediated by AAC(6\u2032)-Ib in vitro. This action seems to be independent of other mechanisms, like increased production of reactive oxygen species and enhanced membrane permeability, proposed to explain the potentiation of the antibiotic effect by silver ions. The addition of this compound to aac(6\u2032)-Ib harboring Acinetobacter baumannii and Escherichia coli cultures resulted in a dramatic reduction of the resistance levels. Time-kill assays showed that the combination of silver acetate and amikacin was bactericidal and exhibited low cytotoxicity to HEK293 cells.<\/jats:p>","DOI":"10.3390\/antibiotics10010029","type":"journal-article","created":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T10:10:37Z","timestamp":1609409437000},"page":"29","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Aminoglycoside 6\u2032-N-acetyltransferase Type Ib [AAC(6\u2032)-Ib]-Mediated Aminoglycoside Resistance: Phenotypic Conversion to Susceptibility by Silver Ions"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0769-395X","authenticated-orcid":false,"given":"Craig M.","family":"Reeves","sequence":"first","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesus","family":"Magallon","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth","family":"Rocha","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tung","family":"Tran","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kimberly","family":"Phan","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Vu","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Yi","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Crista L.","family":"Oakley-Havens","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9","family":"Cedano","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0744-8137","authenticated-orcid":false,"given":"Ver\u00f3nica","family":"Jimenez","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9904-7890","authenticated-orcid":false,"given":"Maria S.","family":"Ramirez","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6298-7811","authenticated-orcid":false,"given":"Marcelo E.","family":"Tolmasky","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology Studies, Department of Biological Science, California State University Fullerton, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Barras, F., Aussel, L., and Ezraty, B. (2018). Silver and antibiotic, new facts to an old Story. Antibiotics, 7.","DOI":"10.3390\/antibiotics7030079"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1007\/s10534-013-9645-z","article-title":"Antimicrobial silver: Uses, toxicity and potential for resistance","volume":"26","author":"Mijnendonckx","year":"2013","journal-title":"Biometals"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5471","DOI":"10.1038\/s41598-019-42049-5","article-title":"Antibacterial activity of combinatorial treatments composed of transition-metal\/antibiotics against Mycobacterium tuberculosis","volume":"9","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_4","first-page":"190ra181","article-title":"Silver enhances antibiotic activity against gram-negative bacteria","volume":"5","author":"Winkler","year":"2013","journal-title":"Sci. Transl. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1038\/s41598-017-01017-7","article-title":"Synergistic antimicrobial effects of silver\/transition-metal combinatorial treatments","volume":"7","author":"Castillo","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7281","DOI":"10.1038\/s41598-020-64127-9","article-title":"Antimicrobial activity of a silver-microfibrillated cellulose biocomposite against susceptible and resistant bacteria","volume":"10","author":"Dugmore","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1111\/mmi.13687","article-title":"Silver potentiates aminoglycoside toxicity by enhancing their uptake","volume":"105","author":"Herisse","year":"2017","journal-title":"Mol. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1016\/j.idc.2016.08.001","article-title":"Multidrug-resistant Gram-negative bacilli: Infection control implications","volume":"30","author":"Adler","year":"2016","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1086\/595011","article-title":"Bad bugs, no drugs: No ESKAPE! An update from the Infectious Diseases Society of America","volume":"48","author":"Boucher","year":"2009","journal-title":"Clin. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1016\/S0002-9343(88)80233-X","article-title":"Multiple intensive care unit outbreak of Acinetobacter calcoaceticus subspecies anitratus respiratory infection and colonization associated with contaminated, reusable ventilator circuits and resuscitation bags","volume":"85","author":"Hartstein","year":"1988","journal-title":"Am. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1111\/j.1574-6976.2012.00344.x","article-title":"Acinetobacter baumannii: Human infections, factors contributing to pathogenesis and animal models","volume":"37","author":"McConnell","year":"2013","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ramirez, M.S., Bonomo, R.A., and Tolmasky, M.E. (2020). Carbapenemases: Transforming Acinetobacter baumannii into a yet more dangerous menace. Biomolecules, 10.","DOI":"10.3390\/biom10050720"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1186\/cc5911","article-title":"Attributable mortality of Acinetobacter baumannii: No longer a controversial issue","volume":"11","author":"Falagas","year":"2007","journal-title":"Crit. Care"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1038\/nrmicro1789","article-title":"An increasing threat in hospitals: Multidrug-resistant Acinetobacter baumannii","volume":"5","author":"Dijkshoorn","year":"2007","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_15","unstructured":"CDC (2019). Antibiotic Resistance Threats in the United States, Centers for Disease Control."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"318","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":"ref_17","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1097\/01.qco.0000185985.64759.41","article-title":"Treatment of multidrug resistant Acinetobacter","volume":"18","author":"Murray","year":"2005","journal-title":"Curr. Opin. Infect. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Ramirez, M.S., and Tolmasky, M.E. (2017). Amikacin: Uses, resistance, and prospects for inhibition. Molecules, 22.","DOI":"10.3390\/molecules22122267"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Kumar, P., Bhardwaj, T., Kumar, A., Gehi, B.R., Kapuganti, S.K., Garg, N., Nath, G., and Giri, R. (2020). Reprofiling of approved drugs against SARS-CoV-2 main protease: An in-silico study. J. Biomol. Struct. Dyn., 1\u201315.","DOI":"10.1080\/07391102.2020.1845976"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.drup.2010.08.003","article-title":"Aminoglycoside modifying enzymes","volume":"13","author":"Ramirez","year":"2010","journal-title":"Drug Resist. Updates"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"121","DOI":"10.3389\/fmicb.2013.00121","article-title":"Rise and dissemination of aminoglycoside resistance: The aac(6\u2019)-Ib paradigm","volume":"4","author":"Ramirez","year":"2013","journal-title":"Front. Microbiol."},{"key":"ref_22","first-page":"1","article-title":"Strategies to prolong the useful life of existing antibiotics and help overcoming the antibiotic resistance crisis","volume":"Volume 1","author":"Rhaman","year":"2017","journal-title":"Frontiers in Clinical Drug Research-Anti Infectives"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.4155\/fmc.13.80","article-title":"Strategies to overcome the action of aminoglycoside-modifying enzymes for treating resistant bacterial infections","volume":"5","author":"Labby","year":"2013","journal-title":"Future Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1038\/s41579-019-0159-8","article-title":"Interplay between beta-lactamases and new beta-lactamase inhibitors","volume":"17","author":"Bush","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5851","DOI":"10.1128\/AAC.01106-15","article-title":"Inhibition of aminoglycoside 6\u2032-N-acetyltransferase type Ib-mediated amikacin resistance in Klebsiella pneumoniae by zinc and copper pyrithione","volume":"59","author":"Chiem","year":"2015","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.ijantimicag.2017.08.002","article-title":"Inhibition of aminoglycoside 6\u2032-N-acetyltransferase type Ib-mediated amikacin resistance by zinc complexed with clioquinol, an ionophore active against tumors and neurodegenerative diseases","volume":"51","author":"Chiem","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4238","DOI":"10.1128\/AAC.00129-14","article-title":"Inhibition of aminoglycoside 6\u2019-N-acetyltransferase type Ib by zinc: Reversal of amikacin resistance in Acinetobacter baumannii and Escherichia coli by a zinc ionophore","volume":"58","author":"Lin","year":"2014","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Magallon, J., Chiem, K., Tran, T., Ramirez, M.S., Jimenez, V., and Tolmasky, M.E. (2019). Restoration of susceptibility to amikacin by 8-hydroxyquinoline analogs complexed to zinc. PLoS ONE, 14.","DOI":"10.1101\/598912"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1016\/j.ijantimicag.2018.01.019","article-title":"Identification of a small molecule inhibitor of the aminoglycoside 6\u2032-N-acetyltransferase type Ib [AAC(6\u2032)-Ib] using mixture-based combinatorial libraries","volume":"51","author":"Tran","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2107","DOI":"10.1002\/cbic.201800368","article-title":"Cellular studies of an aminoglycoside potentiator reveal a new inhibitor of aminoglycoside resistance","volume":"19","author":"Guan","year":"2018","journal-title":"Chembiochem"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1021\/cb200366u","article-title":"Inhibitors of aminoglycoside resistance activated in cells","volume":"7","author":"Vong","year":"2012","journal-title":"ACS Chem. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.diagmicrobio.2013.10.001","article-title":"Increased prevalence of aminoglycoside resistance in clinical isolates of Escherichia coli and Klebsiella spp. in Norway is associated with the acquisition of AAC(3)-II and AAC(6\u2032)-Ib","volume":"78","author":"Haldorsen","year":"2014","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1039\/C5MD00316D","article-title":"Identification of an inhibitor of the aminoglycoside 6\u2032-N-acetyltransferase type Ib [AAC(6\u2032)-Ib] by glide molecular docking","volume":"7","author":"Chiem","year":"2016","journal-title":"Medchemcomm"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5694","DOI":"10.1016\/j.bmcl.2013.08.016","article-title":"Inhibitors of the aminoglycoside 6\u2032-N-acetyltransferase type Ib [AAC(6\u2032)-Ib] identified by in silico molecular docking","volume":"23","author":"Lin","year":"2013","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1368","DOI":"10.1002\/cbic.200700677","article-title":"NMR-guided fragment-based approach for the design of AAC(6\u2032)-Ib ligands","volume":"9","author":"Lombes","year":"2008","journal-title":"Chembiochem"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e02391-18","DOI":"10.1128\/mBio.02391-18","article-title":"Chemical synergy between ionophore PBT2 and zinc reverses antibiotic resistance","volume":"9","author":"Bohlmann","year":"2018","journal-title":"mBio"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4148","DOI":"10.1128\/AAC.00885-15","article-title":"Inhibition of aminoglycoside acetyltransferase resistance enzymes by metal salts","volume":"59","author":"Li","year":"2015","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2323","DOI":"10.1016\/j.ccr.2005.04.027","article-title":"Chemical and biological aspects of Cu2+ interactions with peptides and aminoglycosides","volume":"249","author":"Kozlowski","year":"2005","journal-title":"Coordin Chem Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/jac\/dkn279","article-title":"Functional characterization of Tn1331 gene cassettes","volume":"62","author":"Ramirez","year":"2008","journal-title":"J. Antimicrob. Chemother"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Vilacoba, E., Deraspe, M., Traglia, G.M., Roy, P.H., Ramirez, M.S., and Centron, D. (2014). Draft genome sequence of an international clonal lineage 1 Acinetobacter baumannii strain from Argentina. Genome Announc., 2.","DOI":"10.1128\/genomeA.01190-14"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e00212-15","DOI":"10.1128\/genomeA.00212-15","article-title":"Draft genome of the multidrug-resistant Acinetobacter baumannii strain A155 clinical isolate","volume":"3","author":"Arivett","year":"2015","journal-title":"Genome Announc."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1093\/nar\/gkt918","article-title":"High-copy bacterial plasmids diffuse in the nucleoid-free space, replicate stochastically and are randomly partitioned at cell division","volume":"42","author":"Tran","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3422","DOI":"10.1128\/AAC.46.11.3422-3427.2002","article-title":"Complete nucleotide sequence of Klebsiella pneumoniae multiresistance plasmid pJHCMW1","volume":"46","author":"Sarno","year":"2002","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1955","DOI":"10.1128\/AAC.31.12.1955","article-title":"Tn1331, a novel multiresistance transposon encoding resistance to amikacin and ampicillin in Klebsiella pneumoniae","volume":"31","author":"Tolmasky","year":"1987","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2110","DOI":"10.1073\/pnas.69.8.2110","article-title":"Nonchromosomal antibiotic resistance in bacteria: Genetic transformation of Escherichia coli by R-factor DNA","volume":"69","author":"Cohen","year":"1972","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1488","DOI":"10.1128\/JCM.01264-09","article-title":"Naturally competent Acinetobacter baumannii clinical isolate as a convenient model for genetic studies","volume":"48","author":"Ramirez","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1016\/0076-6879(75)43124-X","article-title":"Aminoglycoside-modifying enzymes","volume":"43","author":"Haas","year":"1975","journal-title":"Methods Enzymol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1128\/AAC.30.2.315","article-title":"Molecular cloning of amikacin resistance determinants from a Klebsiella pneumoniae plasmid","volume":"30","author":"Tolmasky","year":"1986","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1128\/AAC.29.2.315","article-title":"Plasmid-encoded amikacin resistance in multiresistant strains of Klebsiella pneumoniae isolated from neonates with meningitis","volume":"29","author":"Woloj","year":"1986","journal-title":"Antimicrob. Agents Chemother"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/0006-2952(61)90145-9","article-title":"A new and rapid colorimetric determination of acetylcholinesterase activity","volume":"7","author":"Ellman","year":"1961","journal-title":"Biochem. Pharmacol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1099\/0022-1317-36-1-59","article-title":"Characteristics of a human cell line transformed by DNA from human adenovirus type 5","volume":"36","author":"Graham","year":"1977","journal-title":"J. Gen. Virol."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/1\/29\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:48:33Z","timestamp":1760179713000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/1\/29"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,31]]},"references-count":51,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["antibiotics10010029"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics10010029","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12,31]]}}}