{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T14:53:20Z","timestamp":1782485600504,"version":"3.54.5"},"reference-count":45,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,31]],"date-time":"2025-05-31T00:00:00Z","timestamp":1748649600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002923","name":"CONICET","doi-asserted-by":"publisher","award":["PIP 11220200102513CO"],"award-info":[{"award-number":["PIP 11220200102513CO"]}],"id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002923","name":"CONICET","doi-asserted-by":"publisher","award":["UBA (20020220400062BA)"],"award-info":[{"award-number":["UBA (20020220400062BA)"]}],"id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Respiratory viruses can cause life-threatening conditions such as sepsis and acute respiratory distress syndrome. However, vaccines and effective antivirals are available for only a limited number of infections. The majority of approved antivirals are direct-acting agents, which target viral proteins essential for infection. Unfortunately, mutations have already emerged that confer resistance to these antivirals. In addition, there is an urgent need for broad-spectrum antivirals to address the unpredictable emergence of new viruses with pandemic potential. One promising strategy involves modulating the innate immune response and cellular signaling. Imiquimod, a Toll-like receptor 7 (TLR7) agonist, has shown efficacy in murine models of influenza and respiratory syncytial virus (RSV). Additionally, it demonstrates antiviral activity against herpes simplex virus type 1 (HSV-1) and RSV independent of the TLR7\/nuclear factor kappa B (NF-\u03baB) pathway, with protein kinase A (PKA) as a crucial downstream effector. In this study, we demonstrate that imiquimod exhibits concentration-dependent antiviral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and canine coronavirus (CCoV) in epithelial cells, underscoring its broad-spectrum action against coronaviruses. Moreover, its anti-coronavirus effect appears to be independent of the TLR\/NF-\u03baB and PKA\/exchange protein directly activated by cyclic adenosine monophosphate (EPAC) pathways and may instead be linked to the activation of the mitogen-activated protein kinase kinase (MEK)\/extracellular signal-regulated kinase (ERK) pathway. The ability of imiquimod to inhibit coronavirus replication via the MEK\/ERK pathway, coupled with its immunomodulatory properties, highlights its potential as a broad-spectrum antiviral.<\/jats:p>","DOI":"10.3390\/v17060801","type":"journal-article","created":{"date-parts":[[2025,6,2]],"date-time":"2025-06-02T12:27:09Z","timestamp":1748867229000},"page":"801","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Imiquimod, a Promising Broad-Spectrum Antiviral, Prevents SARS-CoV-2 and Canine Coronavirus Multiplication Through the MAPK\/ERK Signaling Pathway"],"prefix":"10.3390","volume":"17","author":[{"given":"Josefina","family":"Vicente","sequence":"first","affiliation":[{"name":"Laboratorio de Virolog\u00eda, Departamento de Qu\u00edmica Biol\u00f3gica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires 1428, Argentina"},{"name":"Instituto de Qu\u00edmica Biol\u00f3gica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET\u2014Universidad de Buenos Aires, Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Freddy Armando","family":"Pe\u00f1aranda Figueredo","sequence":"additional","affiliation":[{"name":"Laboratorio de Virolog\u00eda, Departamento de Qu\u00edmica Biol\u00f3gica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires 1428, Argentina"},{"name":"Instituto de Qu\u00edmica Biol\u00f3gica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET\u2014Universidad de Buenos Aires, Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5885-2842","authenticated-orcid":false,"given":"Stefania","family":"Mantovani","sequence":"additional","affiliation":[{"name":"Division of Clinical Immunology and Infectious Diseases, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela Laura","family":"Papademetrio","sequence":"additional","affiliation":[{"name":"C\u00e1tedra de Inmunolog\u00eda, Departamento de Microbiolog\u00eda, Inmunolog\u00eda, Biotecnolog\u00eda y Gen\u00e9tica, Facultad de Farmacia y Bioqu\u00edmica, Universidad de Buenos Aires, Buenos Aires 1113, Argentina"},{"name":"Unidad de Conocimiento Traslacional, Hospital de Alta Complejidad del Bicentenario Esteban Echeverr\u00eda, Monte Grande, Buenos Aires 1842, Argentina"},{"name":"Centro de Investigaciones en Biomedicina Traslacional CIBiMeT, CONICET\u2014Hospital de Alta Complejidad del Bicentenario Esteban Echeverr\u00eda, Monte Grande, Buenos Aires 1842, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4501-7052","authenticated-orcid":false,"given":"Sergio Ivan","family":"Nemirovsky","sequence":"additional","affiliation":[{"name":"Instituto de Qu\u00edmica Biol\u00f3gica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET\u2014Universidad de Buenos Aires, Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea Alejandra","family":"Barquero","sequence":"additional","affiliation":[{"name":"Laboratorio de Virolog\u00eda, Departamento de Qu\u00edmica Biol\u00f3gica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires 1428, Argentina"},{"name":"Instituto de Qu\u00edmica Biol\u00f3gica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET\u2014Universidad de Buenos Aires, Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carina","family":"Shayo","sequence":"additional","affiliation":[{"name":"Laboratorio de Patolog\u00eda y Farmacolog\u00eda Molecular, Instituto de Biolog\u00eda y Medicina Experimental (IBYME, CONICET), Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0209-6870","authenticated-orcid":false,"given":"Carlos Alberto","family":"Bueno","sequence":"additional","affiliation":[{"name":"Laboratorio de Virolog\u00eda, Departamento de Qu\u00edmica Biol\u00f3gica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires 1428, Argentina"},{"name":"Instituto de Qu\u00edmica Biol\u00f3gica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET\u2014Universidad de Buenos Aires, Buenos Aires 1428, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,31]]},"reference":[{"key":"ref_1","unstructured":"Ryan, E.T., Hill, D.R., Solomon, T., Aronson, N.E., and Endy, T.P. (2020). Viral Respiratory Infections. Hunter\u2019s Tropical Medicine and Emerging Infectious Diseases, Elsevier. [10th ed.]."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Muthukutty, P., MacDonald, J., and Yoo, S.Y. (2024). Combating Emerging Respiratory Viruses: Lessons and Future Antiviral Strategies. Vaccines, 12.","DOI":"10.3390\/vaccines12111220"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1038\/s41579-021-00542-7","article-title":"Influenza virus and SARS-CoV-2: Pathogenesis and host responses in the respiratory tract","volume":"19","author":"Flerlage","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","first-page":"111","article-title":"2019 update of the drug resistance mutations in HIV-1","volume":"27","author":"Wensing","year":"2019","journal-title":"Top. Antivir. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"eabo0718","DOI":"10.1126\/scitranslmed.abo0718","article-title":"Mutations in the SARS-CoV-2 RNA-dependent RNA polymerase confer resistance to remdesivir by distinct mechanisms","volume":"14","author":"Stevens","year":"2022","journal-title":"Sci. Transl. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1038\/s41586-022-05514-2","article-title":"Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir","volume":"613","author":"Iketani","year":"2023","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3604","DOI":"10.1038\/s41467-024-47941-x","article-title":"Dynamic diversity of SARS-CoV-2 genetic mutations in a lung transplantation patient with persistent COVID-19","volume":"15","author":"Igari","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2400786","DOI":"10.2807\/1560-7917.ES.2024.29.50.2400786","article-title":"An outbreak of influenza A(H1N1)pdm09 antigenic variants exhibiting cross-resistance to oseltamivir and peramivir in an elementary school in Japan, September 2024","volume":"29","author":"Takashita","year":"2024","journal-title":"Euro Surveill."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2469643","DOI":"10.1080\/22221751.2025.2469643","article-title":"Neuraminidase reassortment and oseltamivir resistance in clade 2.3.4.4b A(H5N1) viruses circulating among Canadian poultry, 2024","volume":"14","author":"Signore","year":"2025","journal-title":"Emerg. Microbes Infect."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Maffei, M.E., Salata, C., and Gribaudo, G. (2022). Tackling the future pandemics: Broad-spectrum antiviral agents (BSAAs) based on A-type proanthocyanidins. Molecules, 27.","DOI":"10.3390\/molecules27238353"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1126\/science.abn1900","article-title":"Stopping pandemics before they start: Lessons learned from SARS-CoV-2","volume":"375","author":"Edwards","year":"2022","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e170236","DOI":"10.1172\/JCI170236","article-title":"Preparing for the next viral threat with broad-spectrum antivirals","volume":"133","author":"Karim","year":"2023","journal-title":"J. Clin. Investig."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1016\/j.chembiol.2022.05.009","article-title":"Identifying enhancers of innate immune signaling as broad-spectrum antivirals active against emerging viruses","volume":"29","author":"Maarifi","year":"2022","journal-title":"Cell Chem. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e00168-19","DOI":"10.1128\/CMR.00168-19","article-title":"Host-Directed Antiviral Therapy","volume":"33","author":"Kumar","year":"2020","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Latreille, E., and Lee, W.L. (2023). Modulation of the host response as a therapeutic strategy in severe lung infections. Viruses, 15.","DOI":"10.3390\/v15071462"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1016\/j.virol.2015.02.040","article-title":"The role of PI3K\/Akt in human herpesvirus infection: From the bench to the bedside","volume":"479\u2013480","author":"Liu","year":"2015","journal-title":"Virology"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Liu, S., Qiu, F., Gu, R., and Xu, E. (2024). Functional Involvement of Signal Transducers and Activators of Transcription in the Pathogenesis of Influenza A Virus. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252413589"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Liu, Y., and Luo, Z. (2024). Repurposing Anticancer Drugs Targeting the MAPK\/ERK Signaling Pathway for the Treatment of Respiratory Virus Infections. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25136946"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Ba, J., Luan, J., Qi, Z., and Liu, B. (2025). Orthoflavivirus infection and the mTOR signaling pathway. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1565350"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1517\/14728222.10.1.69","article-title":"The small-molecule immune response modifier imiquimod\u2014Its mode of action and clinical use in the treatment of skin cancer","volume":"10","year":"2006","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"To, E.E., Erlich, J., Liong, F., Luong, R., Liong, S., Bozinovski, S., Seow, H.J., O\u2019Leary, J.J., Brooks, D.A., and Vlahos, R. (2019). Intranasal and epicutaneous administration of Toll-like receptor 7 (TLR7) agonists provides protection against influenza A virus-induced morbidity in mice. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-38864-5"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"104817","DOI":"10.1016\/j.antiviral.2020.104817","article-title":"Imiquimod suppresses respiratory syncytial virus (RSV) replication via PKA pathway and reduces RSV inducedinflammation and viral load in mice lungs","volume":"179","author":"Salinas","year":"2020","journal-title":"Antivir. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"10338","DOI":"10.1128\/JVI.01196-12","article-title":"Imiquimod Suppresses Propagation of Herpes Simplex Virus 1 by Upregulation of Cystatin A via the Adenosine Receptor A1 Pathway","volume":"86","author":"Kan","year":"2012","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Vicente, J., Benedetti, M., Martelliti, P., V\u00e1zquez, L., Gentilini, M.V., Figueredo, F.A.P., Jodar, M.S.N., Viegas, M., Barquero, A.A., and Bueno, C.A. (2023). The Flavonoid Cyanidin Shows Immunomodulatory and Broad-Spectrum Antiviral Properties, Including SARS-CoV-2. Viruses, 15.","DOI":"10.3390\/v15040989"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1111\/jphp.13002","article-title":"Virucidal, antiviral and immunomodulatory activities of \u03b2-escin and Aesculus hippocastanum extract","volume":"70","author":"Michelini","year":"2018","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Figueredo, F.A.P., Vicente, J., Barquero, A.A., and Bueno, C.A. (2024). Aesculus hippocastanum extract and the main bioactive constituent \u03b2-escin as antivirals agents against coronaviruses, including SARS-CoV-2. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-56759-y"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.antiviral.2019.01.018","article-title":"Aesculus hippocastanum L. seed extract shows virucidal and antiviral activities against respiratory syncytial virus (RSV) and reduces lung inflammation in vivo","volume":"164","author":"Salinas","year":"2019","journal-title":"Antivir. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1038\/s41435-021-00157-1","article-title":"Rare variants in Toll-like receptor 7 results in functional impairment and downregulation of cytokine-mediated signaling in COVID-19 patients","volume":"23","author":"Mantovani","year":"2021","journal-title":"Genes Immun."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Boulton, S., Crupi, M.J.F., Singh, S., Carter-Timofte, M.E., Azad, T., Organ, B.C., He, X., Gill, R., Neault, S., and Jamieson, T. (2023). Inhibition of exchange proteins directly activated by cAMP as a strategy for broad-spectrum antiviral development. J. Biol. Chem., 299.","DOI":"10.1016\/j.jbc.2023.104749"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3902","DOI":"10.1128\/JVI.03001-13","article-title":"Blocking of exchange proteins directly activated by cAMP leads to reduced replication of Middle East respiratory syndrome coronavirus","volume":"88","author":"Tao","year":"2014","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"111679","DOI":"10.1016\/j.intimp.2024.111679","article-title":"Unconjugated bilirubin and its derivative ameliorate IMQ-induced psoriasis-like skin inflammation in mice by inhibiting MMP9 and MAPK pathway","volume":"130","author":"Bharatha","year":"2024","journal-title":"Int. Immunopharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Brunetti, J.E., Foscaldi, S., Quintana, V.M., Scolaro, L.A., L\u00f3pez, N., and Castilla, V. (2018). Role of the ERK1\/2 Signaling Pathway in the Replication of Jun\u00edn and Tacaribe Viruses. Viruses, 10.","DOI":"10.3390\/v10040199"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"120092","DOI":"10.1016\/j.lfs.2021.120092","article-title":"ERK and p38 MAPK inhibition controls NF-E2 degradation and profibrotic signaling in renal proximal tubule cells","volume":"287","author":"Li","year":"2021","journal-title":"Life Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s00018-021-04085-1","article-title":"The MEK1\/2-inhibitor ATR-002 efficiently blocks SARS-CoV-2 propagation and alleviates pro-inflammatory cytokine\/chemokine responses","volume":"79","author":"Schreiber","year":"2022","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1016\/S2213-2600(22)00213-2","article-title":"Early treatment to prevent progression of SARS-CoV-2 infection","volume":"10","author":"Cohen","year":"2022","journal-title":"Lancet Respir. Med."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"D\u2019Abramo, A., Vita, S., and Nicastri, E. (2022). The unmet need for COVID-19 treatment in immunocompromised patients. BMC Infect. Dis., 22.","DOI":"10.1186\/s12879-022-07918-x"},{"key":"ref_37","unstructured":"WHO (2025, April 15). Therapeutics and COVID-19: Living Guideline (v13.1). Available online: https:\/\/app.magicapp.org\/#\/guideline\/7789."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Nieto-Fontarigo, J.J., Tillgren, S., Cerps, S., Sverrild, A., Hvidtfeldt, M., Ramu, S., Menzel, M., Sander, A.F., Porsbjerg, C., and Uller, L. (2021). Imiquimod Boosts Interferon Response, and Decreases ACE2 and Pro-Inflammatory Response of Human Bronchial Epithelium in Asthma. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.743890"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1159\/000518471","article-title":"Interferon and Toll-Like Receptor 7 Response in COVID-19: Implications of Topical Imiquimod for Prophylaxis and Treatment","volume":"237","author":"Szeto","year":"2021","journal-title":"Dermatology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"D246","DOI":"10.1093\/nar\/gkx1158","article-title":"Expression Atlas: Gene and protein expression across multiple studies and organisms","volume":"46","author":"Papatheodorou","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e27","DOI":"10.4142\/jvs.21225","article-title":"Expression of Toll-like receptors 3, 7, 9 and cytokines in feline infectious peritonitis virus-infected CRFK cells and feline peripheral monocytes","volume":"23","author":"Khair","year":"2022","journal-title":"J. Vet. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1338","DOI":"10.1038\/sj.jid.5700286","article-title":"The small antitumoral immune response modifier imiquimod interacts with adenosine receptor signaling in a TLR7- and TLR8-independent fashion","volume":"126","author":"Klotz","year":"2006","journal-title":"J. Investig. Dermatol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"9","DOI":"10.2174\/2213988501307010009","article-title":"Minireview: Targeting GPCR Activated ERK Pathways for Drug Discovery","volume":"7","author":"Eishingdrelo","year":"2013","journal-title":"Curr. Chem. Genom. Transl. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"173619","DOI":"10.1016\/j.ejphar.2020.173619","article-title":"Adenosine receptor signalling: Probing the potential pathways for the ministration of neuropathic pain","volume":"889","author":"Shaw","year":"2020","journal-title":"Eur. J. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/bs.acr.2018.02.004","article-title":"Extracellular-Regulated Kinases: Signaling from Ras to ERK Substrates to Control Biological Outcomes","volume":"138","author":"Eblen","year":"2018","journal-title":"Adv. Cancer Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/6\/801\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:44:35Z","timestamp":1760031875000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/6\/801"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,31]]},"references-count":45,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["v17060801"],"URL":"https:\/\/doi.org\/10.3390\/v17060801","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,31]]}}}