{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,11]],"date-time":"2026-01-11T05:34:23Z","timestamp":1768109663344,"version":"3.49.0"},"reference-count":43,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,22]],"date-time":"2022-10-22T00:00:00Z","timestamp":1666396800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda (CONACYT)","doi-asserted-by":"publisher","award":["168896"],"award-info":[{"award-number":["168896"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda (CONACYT)","doi-asserted-by":"publisher","award":["261875"],"award-info":[{"award-number":["261875"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Alterations in DNA methylation are critical for the carcinogenesis of ovarian tumors, especially ovarian carcinoma (OC). DNMT3B, a de novo DNA methyltransferase (DNMT), encodes for fifteen spliced protein products or isoforms. DNMT3B isoforms lack exons for the catalytic domain, with functional consequences on catalytic activity. Abnormal expression of DNMT3B isoforms is frequently observed in several types of cancer, such as breast, lung, kidney, gastric, liver, skin, leukemia, and sarcoma. However, the expression patterns and consequences of DNMT3B isoforms in OC are unknown. In this study, we analyzed each DNMT and DNMT3B isoforms expression by qPCR in 63 OC samples and their association with disease-free survival (DFS), overall survival (OS), and tumor progression. We included OC patients with the main histological subtypes of EOC and patients in all the disease stages and found that DNMTs were overexpressed in advanced stages (p-value &lt; 0.05) and high-grade OC (p-value &lt; 0.05). Remarkably, we found DNMT3B1 overexpression in advanced stages (p-value = 0.0251) and high-grade serous ovarian carcinoma (HGSOC) (p-value = 0.0313), and DNMT3B3 was overexpressed in advanced stages (p-value = 0.0098) and high-grade (p-value = 0.0004) serous ovarian carcinoma (SOC). Finally, we observed that overexpression of DNMT3B isoforms was associated with poor prognosis in OC and SOC. DNMT3B3 was also associated with FDS (p-value = 0.017) and OS (p-value = 0.038) in SOC patients. In addition, the ovarian carcinoma cell lines OVCAR3 and SKOV3 also overexpress DNMT3B3. Interestingly, exogenous overexpression of DNMT3B3 in OVCAR3 causes demethylation of satellite 2 sequences in the pericentromeric region. In summary, our results suggest that DNMT3B3 expression is altered in OC.<\/jats:p>","DOI":"10.3390\/ijms232112759","type":"journal-article","created":{"date-parts":[[2022,10,24]],"date-time":"2022-10-24T04:40:36Z","timestamp":1666586436000},"page":"12759","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Expression of DNA Methyltransferase 3B Isoforms Is Associated with DNA Satellite 2 Hypomethylation and Clinical Prognosis in Advanced High-Grade Serous Ovarian Carcinoma"],"prefix":"10.3390","volume":"23","author":[{"given":"Victor M.","family":"Del Castillo Falconi","sequence":"first","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9583-3790","authenticated-orcid":false,"given":"Jos\u00e9","family":"D\u00edaz-Ch\u00e1vez","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4658-5310","authenticated-orcid":false,"given":"Karla","family":"Torres-Arciga","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fernando","family":"Luna-Maldonado","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adriana A.","family":"Gudi\u00f1o-Gomez","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Medicina Gen\u00f3mica, Periferico Sur 4809, Arenal Tepepan, Tlalpan, Ciudad de M\u00e9xico 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7857-3464","authenticated-orcid":false,"given":"Abraham","family":"Pedroza-Torres","sequence":"additional","affiliation":[{"name":"C\u00e1tedras CONACYT\u2014Instituto Nacional de Cancerolog\u00eda, Ciudad de M\u00e9xico 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5641-3627","authenticated-orcid":false,"given":"Clementina","family":"Castro-Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Cant\u00fa de Le\u00f3n","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3998-9306","authenticated-orcid":false,"given":"Luis A.","family":"Herrera","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Cancerolog\u00eda (INCAN), Unidad de Investigaci\u00f3n Biom\u00e9dica en C\u00e1ncer, Instituto de Investigaciones Biom\u00e9dicas, UNAM, Avenida San Fernando No. 22, Secci\u00f3n XVI Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"},{"name":"Instituto Nacional de Medicina Gen\u00f3mica, Periferico Sur 4809, Arenal Tepepan, Tlalpan, Ciudad de M\u00e9xico 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"394","DOI":"10.3322\/caac.21492","article-title":"Global Cancer Statistics 2018: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1097\/PAS.0b013e3181cf3d79","article-title":"The Origin and Pathogenesis of Epithelial Ovarian Cancer: A Proposed Unifying Theory","volume":"34","author":"Kurman","year":"2010","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1097\/PAT.0b013e328348a6e7","article-title":"Morphological Subtypes of Ovarian Carcinoma: A Review with Emphasis on New Developments and Pathogenesis","volume":"43","author":"McCluggage","year":"2011","journal-title":"Pathology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"a019133","DOI":"10.1101\/cshperspect.a019133","article-title":"DNA Methylation in Mammals","volume":"6","author":"Li","year":"2014","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/cbic.201000195","article-title":"Structure and Function of Mammalian DNA Methyltransferases","volume":"12","author":"Jurkowska","year":"2011","journal-title":"ChemBioChem"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"R110","DOI":"10.1186\/gb-2012-13-11-r110","article-title":"Human Genes with CpG Island Promoters Have a Distinct Transcription-Associated Chromatin Organization","volume":"13","author":"Vavouri","year":"2012","journal-title":"Genome Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.ygyno.2010.12.332","article-title":"DNA Ploidy, Nuclear Size, Proliferation Index and DNA-Hypomethylation in Ovarian Cancer","volume":"121","author":"Zeimet","year":"2011","journal-title":"Gynecol. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4472","DOI":"10.1158\/0008-5472.CAN-04-0238","article-title":"DNA Hypomethylation and Ovarian Cancer Biology","volume":"64","author":"Widschwendter","year":"2004","journal-title":"Cancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1002\/cncr.21992","article-title":"Aberrant Promoter Methylation of Multiple Genes in Malignant Ovarian Tumors and in Ovarian Tumors with Low Malignant Potential","volume":"107","author":"Wiley","year":"2006","journal-title":"Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"165","DOI":"10.4161\/epi.4.3.8277","article-title":"Hemimethylation Footprints of DNA Demethylation in Cancer","volume":"4","author":"Shao","year":"2009","journal-title":"Epigenetics"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Press, J.Z., De Luca, A., Boyd, N., Young, S., Troussard, A., Ridge, Y., Kaurah, P., Kalloger, S.E., Blood, K.A., and Smith, M. (2008). Ovarian Carcinomas with Genetic and Epigenetic BRCA1 Loss Have Distinct Molecular Abnormalities. BMC Cancer, 8.","DOI":"10.1186\/1471-2407-8-17"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6476","DOI":"10.1158\/0008-5472.CAN-04-1529","article-title":"Tumor Cell-Specific BRCA1 and RASSF1A Hypermethylation in Serum, Plasma, and Peritoneal Fluid from Ovarian Cancer Patients","volume":"64","author":"Battagli","year":"2004","journal-title":"Cancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1092","DOI":"10.3892\/etm.2012.715","article-title":"Methylation of Tumor Suppressor Genes in Ovarian Cancer","volume":"4","author":"Ozdemir","year":"2012","journal-title":"Exp. Ther. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1177\/0748233713484657","article-title":"Hypermethylation of P 15, P 16, and E-Cadherin Genes in Ovarian Cancer","volume":"31","author":"Moselhy","year":"2015","journal-title":"Toxicol. Ind. Health"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/B978-0-12-387685-0.00007-X","article-title":"The DNMT3 Family of Mammalian De Novo DNA Methyltransferases","volume":"Volume 101","year":"2011","journal-title":"Progress in Molecular Biology and Translational Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0092-8674(00)81656-6","article-title":"DNA Methyltransferases Dnmt3a and Dnmt3b Are Essential for De Novo Methylation and Mammalian Development","volume":"99","author":"Okano","year":"1999","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1006\/dbio.2002.0628","article-title":"Dynamics of Dnmt1 Methyltransferase Expression and Intracellular Localization during Oogenesis and Preimplantation Development","volume":"245","author":"Ratnam","year":"2002","journal-title":"Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3421","DOI":"10.15252\/embj.201797038","article-title":"Isoform-specific Localization of DNMT3A Regulates DNA Methylation Fidelity at Bivalent CpG Islands","volume":"36","author":"Manzo","year":"2017","journal-title":"EMBO J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5553","DOI":"10.1038\/sj.onc.1210351","article-title":"Cancer Cells Express Aberrant DNMT3B Transcripts Encoding Truncated Proteins","volume":"26","author":"Ostler","year":"2007","journal-title":"Oncogene"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1340","DOI":"10.1016\/j.ebiom.2015.09.002","article-title":"\u2206 DNMT3B4-Del Contributes to Aberrant DNA Methylation Patterns in Lung Tumorigenesis","volume":"2","author":"Ma","year":"2015","journal-title":"EBioMedicine"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"10647","DOI":"10.1158\/0008-5472.CAN-07-1337","article-title":"\u0394DNMT3B Variants Regulate DNA Methylation in a Promoter-Specific Manner","volume":"67","author":"Wang","year":"2007","journal-title":"Cancer Res."},{"key":"ref_22","first-page":"201","article-title":"A Novel DNMT3B Subfamily, \u0394DNMT3B, Is the Predominant Form of DNMT3B in Non-Small Cell Lung Cancer","volume":"29","author":"Wang","year":"2006","journal-title":"Int. J. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1622","DOI":"10.1158\/1541-7786.MCR-09-0018","article-title":"A Novel DNMT3B Splice Variant Expressed in Tumor and Pluripotent Cells Modulates Genomic DNA Methylation Patterns and Displays Altered DNA Binding","volume":"7","author":"Gopalakrishnan","year":"2009","journal-title":"Mol. Cancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1038\/890","article-title":"Cloning and Characterization of a Family of Novel Mammalian DNA (Cytosine-5) Methyltransferases","volume":"19","author":"Okano","year":"1998","journal-title":"Nat. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1158\/1541-7786.62.2.1","article-title":"Role of the DNA Methyltransferase Variant DNMT3b3 in DNA Methylation","volume":"1","author":"Weisenberger","year":"2004","journal-title":"Mol. Cancer Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3506","DOI":"10.1093\/nar\/29.17.3506","article-title":"Enzymatic Properties of de Novo-Type Mouse DNA (Cytosine-5) Methyltransferases","volume":"29","author":"Aoki","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Gordon, C.A., Hartono, S.R., and Ch\u00e9din, F. (2013). Inactive DNMT3B Splice Variants Modulate De Novo DNA Methylation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069486"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1007\/s11626-013-9619-z","article-title":"Splice Variants DNMT3B4 and DNMT3B7 Overexpression Inhibit Cell Proliferation in 293A Cell Line","volume":"49","author":"Shao","year":"2013","journal-title":"Vitro Cell. Dev. Biol. Anim."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4714","DOI":"10.1158\/0008-5472.CAN-12-0886","article-title":"Truncated DNMT3B Isoform DNMT3B7 Suppresses Growth, Induces Differentiation, and Alters DNA Methylation in Human Neuroblastoma","volume":"72","author":"Ostler","year":"2012","journal-title":"Cancer Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Gopalakrishna-Pillai, S., and Iverson, L.E. (2011). A DNMT3B Alternatively Spliced Exon and Encoded Peptide Are Novel Biomarkers of Human Pluripotent Stem Cells. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0020663"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.2217\/epi-2016-0013","article-title":"Genome-Wide Methylation Analysis of DNMT3B Gene Isoforms Revealed Specific Methylation Profiles in Breast Cell Lines","volume":"8","author":"Plourde","year":"2016","journal-title":"Epigenomics"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"11453","DOI":"10.1038\/ncomms11453","article-title":"DNMT3B Isoforms without Catalytic Activity Stimulate Gene Body Methylation as Accessory Proteins in Somatic Cells","volume":"7","author":"Duymich","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_33","first-page":"819","article-title":"Expression Pattern and Clinical Significance of DNA Methyltransferase 3B Variants in Gastric Carcinoma","volume":"8","author":"Su","year":"2010","journal-title":"Oncol. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"19712","DOI":"10.18632\/oncotarget.14966","article-title":"An Association between Overexpression of DNA Methyltransferase 3B4 and Clear Cell Renal Cell Carcinoma","volume":"8","author":"Liu","year":"2017","journal-title":"Oncotarget"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10060","DOI":"10.1073\/pnas.152121799","article-title":"Overexpression of a Splice Variant of DNA Methyltransferase 3b, DNMT3b4, Associated with DNA Hypomethylation on Pericentromeric Satellite Regions during Human Hepatocarcinogenesis","volume":"99","author":"Saito","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Brambert, P.R., Kelpsch, D.J., Hameed, R., Desai, C.V., Calafiore, G., Godley, L.A., and Raimondi, S.L. (2015). DNMT3B7 Expression Promotes Tumor Progression to a More Aggressive Phenotype in Breast Cancer Cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0117310"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Siddiqui, S., White, M.W., Schroeder, A.M., DeLuca, N.V., Leszczynski, A.L., and Raimondi, S.L. (2018). Aberrant DNMT3B7 Expression Correlates to Tissue Type, Stage, and Survival across Cancers. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0201522"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1006\/gyno.2001.6284","article-title":"DNA Methylation in Ovarian Cancer: II. Expression of DNA Methyltransferases in Ovarian Cancer Cell Lines and Normal Ovarian Epithelial Cells","volume":"82","author":"Ahluwalia","year":"2001","journal-title":"Gynecol. Oncol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4984","DOI":"10.1093\/nar\/gkr116","article-title":"Modulation of Dnmt3b Function in Vitro by Interactions with Dnmt3L, Dnmt3a and Dnmt3b Splice Variants","volume":"39","author":"Robertson","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2291","DOI":"10.1093\/nar\/27.11.2291","article-title":"The Human DNA Methyltransferases (DNMTs) 1, 3a and 3b: Coordinate MRNA Expression in Normal Tissues and Overexpression in Tumors","volume":"27","author":"Robertson","year":"1999","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"8361","DOI":"10.1158\/0008-5472.CAN-06-2031","article-title":"Expression of \u0394DNMT3B Variants and Its Association with Promoter Methylation of P16 and RASSF1A in Primary Non\u2013Small Cell Lung Cancer","volume":"66","author":"Wang","year":"2006","journal-title":"Cancer Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2636","DOI":"10.1038\/sj.onc.1209145","article-title":"Quantitative Analysis of Associations between DNA Hypermethylation, Hypomethylation, and DNMT RNA Levels in Ovarian Tumors","volume":"25","author":"Ehrlich","year":"2006","journal-title":"Oncogene"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/21\/12759\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:01:07Z","timestamp":1760144467000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/21\/12759"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,22]]},"references-count":43,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["ijms232112759"],"URL":"https:\/\/doi.org\/10.3390\/ijms232112759","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,22]]}}}