{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:06:14Z","timestamp":1790791574004,"version":"4.1.0"},"reference-count":53,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2021,11,18]],"date-time":"2021-11-18T00:00:00Z","timestamp":1637193600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"national natural science foundation of china","doi-asserted-by":"publisher","award":["31360593"],"award-info":[{"award-number":["31360593"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Chronic inflammation can cause oviduct mucosal damage and immune dysfunction, leading to infertility, early pregnancy loss, ectopic pregnancy, tumors, and a decrease in reproductive capacities in female animals. Estrogen can suppress immune responses in different tissues and oviducts, and regulate the oviduct immune balance; however, the underlying mechanisms remain unclear. The objective of this study was to explore the mechanism of estrogen-regulated oviduct mucosal immunity and discover new estrogen targets for regulating oviduct mucosal immune homeostasis. Sheep oviduct epithelial cells (SOECs) were treated with 17-\u03b2 estradiol (E2). Transcriptome sequencing and analysis showed differentially expressed S100 calcium-binding protein A (<jats:italic>S100A<\/jats:italic>) genes that may participate in the oviduct mucosa immunoregulation of estrogen. Quantitative polymerase chain reaction and immunocytochemistry analysis showed that<jats:italic>S100A8<\/jats:italic>expression changed dynamically in E2-treated SOECs and peaked after 7 h of treatment. Estrogen nuclear receptors and G protein-coupled membrane receptors promoted E2-dependent<jats:italic>S100A8<\/jats:italic>upregulation. The<jats:italic>S100A8<\/jats:italic>gene was disrupted using the clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR-associated protein 9 method. Levels of inflammatory factors interleukin (IL)-1\u03b2 and IL-4 were significantly upregulated in<jats:italic>S100A8<\/jats:italic>-knockdown SOECs, whereas those of the anti-inflammatory factor IL-10 was downregulated. Following S100A8 knockdown in SOECs treated with E2 for 7 h, IL-10 levels increased significantly. Estrogen affected oviduct mucosa immune function and dynamically regulated S100A8 in SOECs. S100A8 knockdown caused an excessive immune response, indicating that S100A8 is beneficial for maintaining immune homeostasis in the oviduct mucosa. Moreover, estrogen can compensate for the effect of S100A8 knockdown by upregulating IL-10.<\/jats:p>","DOI":"10.1371\/journal.pone.0260188","type":"journal-article","created":{"date-parts":[[2021,11,18]],"date-time":"2021-11-18T18:57:45Z","timestamp":1637261865000},"page":"e0260188","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":11,"title":["S100A8 expression in oviduct mucosal epithelial cells is regulated by estrogen and affects mucosal immune homeostasis"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6507-6810","authenticated-orcid":true,"given":"Xiaodan","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guifang","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxin","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dafu","family":"Zhi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daqing","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moning","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Su","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,11,18]]},"reference":[{"issue":"1","key":"pone.0260188.ref001","doi-asserted-by":"crossref","first-page":"R1","DOI":"10.1530\/JOE-16-0302","article-title":"Oviduct: roles in fertilization and early embryo development","volume":"232","author":"S Li","year":"2017","journal-title":"J Endocrinol"},{"issue":"1","key":"pone.0260188.ref002","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.rvsc.2011.06.023","article-title":"Determination of the distribution of lentogenic vaccine and virulent Newcastle disease virus antigen in the oviduct of SPF and commercial hen using immunohistochemistry","volume":"93","author":"DG Bwala","year":"2012","journal-title":"Res Vet Sci"},{"issue":"10","key":"pone.0260188.ref003","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1111\/imcb.12285","article-title":"Chlamydia-infected macrophages are resistant to azithromycin treatment and are associated with chronic oviduct inflammation and hydrosalpinx development","volume":"97","author":"MC Harvie","year":"2019","journal-title":"Immunol Cell Biol"},{"issue":"1","key":"pone.0260188.ref004","doi-asserted-by":"crossref","first-page":"12394","DOI":"10.1038\/s41598-018-30261-8","article-title":"Inflammation is a key contributor to ovarian cancer cell seeding","volume":"8","author":"D Jia","year":"2018","journal-title":"Sci Rep"},{"issue":"1","key":"pone.0260188.ref005","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.fertnstert.2005.12.033","article-title":"Pregnancy outcome in patients with early recurrent abortion following laparoscopic tubal corneal interruption of a fallopian tube with hydrosalpinx","volume":"86","author":"J Zolghadri","year":"2006","journal-title":"Fertil Steril"},{"issue":"6","key":"pone.0260188.ref006","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1128\/CVI.00006-14","article-title":"Steroids alone or as adjunctive therapy with doxycycline fail to improve oviduct damage in mice infected with Chlamydia muridarum","volume":"21","author":"TE Corr","year":"2014","journal-title":"Clin Vaccine Immunol: CVI"},{"key":"pone.0260188.ref007","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1186\/2046-4053-3-79","article-title":"The effectiveness of nonsteroidal anti-inflammatory agents in the treatment of pelvic inflammatory disease: a systematic review","volume":"3","author":"D Dhasmana","year":"2014","journal-title":"Syst Rev"},{"issue":"7","key":"pone.0260188.ref008","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1093\/carcin\/bgz088","article-title":"Supplemental estrogen and caloric restriction reduce obesity-induced periprostatic white adipose inflammation in mice","volume":"40","author":"P Bhardwaj","year":"2019","journal-title":"Carcinogenesis"},{"issue":"10","key":"pone.0260188.ref009","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.3390\/cells8101259","article-title":"Potential Therapeutic Application of Estrogen in Gender Disparity of Nonalcoholic Fatty Liver Disease\/Nonalcoholic Steatohepatitis","volume":"8","author":"C Lee","year":"2019","journal-title":"Cells"},{"issue":"11","key":"pone.0260188.ref010","doi-asserted-by":"crossref","first-page":"1208","DOI":"10.1097\/GME.0000000000001220","article-title":"17\u03b2-estradiol vaginal tablet versus conjugated equine estrogen vaginal cream to relieve menopausal atrophic vaginitis","volume":"25","author":"JE Rioux","year":"2018","journal-title":"Menopause"},{"issue":"1","key":"pone.0260188.ref011","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1186\/s12974-018-1246-x","article-title":"The novel estrogenic receptor GPR30 alleviates ischemic injury by inhibiting TLR4-mediated microglial inflammation","volume":"15","author":"Z Zhang","year":"2018","journal-title":"J Neuroinflammation"},{"issue":"10","key":"pone.0260188.ref012","doi-asserted-by":"crossref","first-page":"4046","DOI":"10.4049\/jimmunol.1601896","article-title":"Sex-Based Differences in Susceptibility to Severe Acute Respiratory Syndrome Coronavirus Infection","volume":"198","author":"R Channappanavar","year":"2017","journal-title":"J Immunol"},{"issue":"5","key":"pone.0260188.ref013","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1262\/jrd.2013-036","article-title":"Regulation of innate immune function in bovine oviduct epithelial cells in culture: the homeostatic role of epithelial cells in balancing Th1\/Th2 response","volume":"59","author":"R Kowsar","year":"2013","journal-title":"J Reprod Dev"},{"issue":"4","key":"pone.0260188.ref014","doi-asserted-by":"crossref","first-page":"154","DOI":"10.5653\/cerm.2018.45.4.154","article-title":"Sex hormones alter the response of Toll-like receptor 3 to its specific ligand in fallopian tube epithelial cells","volume":"45","author":"Z Zandieh","year":"2018","journal-title":"Clin Exp Reprod Med"},{"key":"pone.0260188.ref015","doi-asserted-by":"crossref","first-page":"e10453","DOI":"10.7554\/eLife.10453","article-title":"Oviductal estrogen receptor \u03b1 signaling prevents protease-mediated embryo death","volume":"4","author":"W Winuthayanon","year":"2015","journal-title":"eLife"},{"issue":"1","key":"pone.0260188.ref016","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.jri.2006.05.001","article-title":"Cytokines in reproductive remodeling of molting White Leghorn hens","volume":"73","author":"NR Sundaresan","year":"2007","journal-title":"J Reprod Immunol"},{"key":"pone.0260188.ref017","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13048-016-0213-3","article-title":"Genome-wide transcriptional regulation of estrogen receptor targets in fallopian tube cells and the role of selective estrogen receptor modulators","volume":"9","author":"G Moyle-Heyrman","year":"2016","journal-title":"J Ovarian Res"},{"issue":"1","key":"pone.0260188.ref018","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s12253-018-00569-x","article-title":"Hormone Replacement Therapy in Cancer Survivors\u2014Review of the Literature","volume":"26","author":"T Deli","year":"2020","journal-title":"Pathol Oncolo Res"},{"issue":"7","key":"pone.0260188.ref019","doi-asserted-by":"crossref","first-page":"e99792","DOI":"10.1371\/journal.pone.0099792","article-title":"Avian SERPINB12 expression in the avian oviduct is regulated by estrogen and up-regulated in epithelial cell-derived ovarian carcinomas of laying hens","volume":"9","author":"G Jo","year":"2014","journal-title":"PloS one"},{"key":"pone.0260188.ref020","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1186\/1746-6148-8-143","article-title":"Modulation of ovine SBD-1 expression by 17beta-estradiol in ovine oviduct epithelial cells","volume":"8","author":"S Wen","year":"2012","journal-title":"BMC Vet Res"},{"issue":"6","key":"pone.0260188.ref021","doi-asserted-by":"crossref","first-page":"3643","DOI":"10.1128\/IAI.00012-06","article-title":"Infection of human fallopian tube epithelial cells with Neisseria gonorrhoeae protects cells from tumor necrosis factor alpha-induced apoptosis","volume":"74","author":"P Morales","year":"2006","journal-title":"Infect Immun"},{"issue":"11","key":"pone.0260188.ref022","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1507\/endocrj.EJ17-0148","article-title":"Rasd1 is an estrogen-responsive immediate early gene and modulates expression of late genes in rat anterior pituitary cells","volume":"64","author":"L Wang","year":"2017","journal-title":"Endocr J"},{"issue":"12","key":"pone.0260188.ref023","first-page":"1605","article-title":"Estrogen exerts anti-inflammatory effects by inhibiting NF-\u03baB pathway through binding with estrogen receptor \u03b2 on synovicytes of osteoarthritis","volume":"32","author":"E Zhang","year":"2016","journal-title":"Chi J Cell Mol Immunol"},{"issue":"3","key":"pone.0260188.ref024","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1080\/08923970600927751","article-title":"In vitro effects of benzophenone-2 and octyl-methoxycinnamate on the production of interferon-gamma and interleukin-10 by murine splenocytes","volume":"28","author":"D Racho\u0144","year":"2006","journal-title":"Immunopharmacol Immunotoxicol"},{"key":"pone.0260188.ref025","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/1471-2261-7-13","article-title":"A cell culture model using rat coronary artery adventitial fibroblasts to measure collagen production","volume":"7","author":"C Jenkins","year":"2007","journal-title":"BMC Cardiovasc Disord"},{"issue":"1","key":"pone.0260188.ref026","doi-asserted-by":"crossref","first-page":"e0147685","DOI":"10.1371\/journal.pone.0147685","article-title":"Estrogen Receptor Alpha (ESR1)-Dependent Regulation of the Mouse Oviductal Transcriptome","volume":"11","author":"KL Cerny","year":"2016","journal-title":"PloS one"},{"issue":"3","key":"pone.0260188.ref027","doi-asserted-by":"crossref","first-page":"1936","DOI":"10.3892\/ol.2016.4134","article-title":"S100A8\/A9 is associated with estrogen receptor loss in breast cancer","volume":"11","author":"YI Bao","year":"2016","journal-title":"Oncol Lett"},{"issue":"3","key":"pone.0260188.ref028","doi-asserted-by":"crossref","first-page":"e13163","DOI":"10.1111\/aji.13163","article-title":"S100A8, which increases with age, induces cellular senescence-like changes in bovine oviduct epithelial cells","volume":"82","author":"Y Nakamura","year":"2019","journal-title":"Am J Reprod Immunol"},{"issue":"9","key":"pone.0260188.ref029","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1002\/mrd.22693","article-title":"Age-dependent changes in inflammation and extracellular matrix in bovine oviduct epithelial cells during the post-ovulatory phase","volume":"83","author":"H Tanaka","year":"2016","journal-title":"Mol Reprod Dev"},{"issue":"5","key":"pone.0260188.ref030","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/icb.2017.2","article-title":"S100A8\/A9 and S100A9 reduce acute lung injury","volume":"95","author":"Y Hiroshima","year":"2017","journal-title":"Immunol Cell Biol"},{"issue":"6","key":"pone.0260188.ref031","doi-asserted-by":"crossref","first-page":"2800","DOI":"10.4049\/jimmunol.1302556","article-title":"S100A8 induces IL-10 and protects against acute lung injury","volume":"192","author":"Y Hiroshima","year":"2014","journal-title":"J Immunol"},{"issue":"6","key":"pone.0260188.ref032","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1038\/ni.3745","article-title":"S100-alarmin-induced innate immune programming protects newborn infants from sepsis","volume":"18","author":"T Ulas","year":"2017","journal-title":"Nat Immunol"},{"issue":"8","key":"pone.0260188.ref033","doi-asserted-by":"crossref","first-page":"e103629","DOI":"10.1371\/journal.pone.0103629","article-title":"TLR9 ligands induce S100A8 in macrophages via a STAT3-dependent pathway which requires IL-10 and PGE2","volume":"9","author":"K Hsu","year":"2014","journal-title":"PloS one"},{"issue":"5","key":"pone.0260188.ref034","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1172\/JCI89867","article-title":"Autoinhibitory regulation of S100A8\/S100A9 alarmin activity locally restricts sterile inflammation","volume":"128","author":"T Vogl","year":"2018","journal-title":"Journal Clin Invest"},{"issue":"1","key":"pone.0260188.ref035","doi-asserted-by":"crossref","first-page":"1685","DOI":"10.1038\/s41467-019-09359-8","article-title":"Impaired cellular energy metabolism in cord blood macrophages contributes to abortive response toward inflammatory threats","volume":"10","author":"S Dreschers","year":"2019","journal-title":"Nat Commun"},{"issue":"2","key":"pone.0260188.ref036","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1002\/jcb.26319","article-title":"Advanced glycation end-products and Porphyromonas gingivalis lipopolysaccharide increase calprotectin expression in human gingival epithelial cells","volume":"119","author":"Y Hiroshima","year":"2018","journal-title":"J Cell Biochem"},{"issue":"9","key":"pone.0260188.ref037","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1161\/CIRCULATIONAHA.118.039262","article-title":"S100a8\/a9 Signaling Causes Mitochondrial Dysfunction and Cardiomyocyte Death in Response to Ischemic\/Reperfusion Injury","volume":"140","author":"Y Li","year":"2019","journal-title":"Circulation"},{"issue":"1","key":"pone.0260188.ref038","doi-asserted-by":"crossref","first-page":"e0169614","DOI":"10.1371\/journal.pone.0169614","article-title":"Expression of Calgranulin Genes S100A8, S100A9 and S100A12 Is Modulated by n-3 PUFA during Inflammation in Adipose Tissue and Mononuclear Cells","volume":"12","author":"RD Shah","year":"2017","journal-title":"PloS one"},{"issue":"2","key":"pone.0260188.ref039","doi-asserted-by":"crossref","first-page":"2475","DOI":"10.18632\/oncotarget.23504","article-title":"Identification of differentially expressed genes in human breast cancer cells induced by 4-hydroxyltamoxifen and elucidation of their pathophysiological relevance and mechanisms","volume":"9","author":"Q Fang","year":"2018","journal-title":"Oncotarget"},{"issue":"2","key":"pone.0260188.ref040","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1111\/aji.12318","article-title":"Association of increased S100A8 serum protein with early pregnancy loss","volume":"73","author":"RR Nair","year":"2015","journal-title":"Am J Reprod Immunol"},{"issue":"8","key":"pone.0260188.ref041","doi-asserted-by":"crossref","first-page":"2493","DOI":"10.1007\/s11250-019-01958-3","article-title":"Targeted transcript analysis revealed association of suboptimal expression of certain endometrial immunity-related genes with disparate uterine diseases in zebu cows","volume":"51","author":"RK Baithalu","year":"2019","journal-title":"Trop Anim Health Prod"},{"issue":"4","key":"pone.0260188.ref042","doi-asserted-by":"crossref","first-page":"2209","DOI":"10.4049\/jimmunol.163.4.2209","article-title":"A null mutation in the inflammation-associated S100 protein S100A8 causes early resorption of the mouse embryo","volume":"163","author":"RJ Passey","year":"1999","journal-title":"J Immunol"},{"key":"pone.0260188.ref043","doi-asserted-by":"crossref","first-page":"20238","DOI":"10.1038\/srep20238","article-title":"Analyses of Long Non-Coding RNA and mRNA profiling using RNA sequencing during the pre-implantation phases in pig endometrium","volume":"6","author":"Y Wang","year":"2016","journal-title":"Sci Rep"},{"key":"pone.0260188.ref044","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/978-3-319-56895-9_6","article-title":"Estrogen Receptors in Cell Membranes: Regulation and Signaling","volume":"227","author":"J Saczko","year":"2017","journal-title":"Adv Anat Embryol Cell Biol"},{"issue":"10","key":"pone.0260188.ref045","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.3390\/cells8101123","article-title":"Estrogen-Receptor Expression and Function in Female Reproductive Disease","volume":"8","author":"ZR Tang","year":"2019","journal-title":"Cells"},{"issue":"3\u20135","key":"pone.0260188.ref046","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/j.jsbmb.2011.07.002","article-title":"Identification of a GPER\/GPR30 antagonist with improved estrogen receptor counterselectivity","volume":"127","author":"MK Dennis","year":"2011","journal-title":"J Steroid Biochem Mol Bio"},{"issue":"7","key":"pone.0260188.ref047","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1038\/s41388-018-0468-9","article-title":"Role of SUMOylation in differential ER\u03b1 transcriptional repression by tamoxifen and fulvestrant in breast cancer cells","volume":"38","author":"T Traboulsi","year":"2019","journal-title":"Oncogene"},{"issue":"11","key":"pone.0260188.ref048","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1111\/1348-0421.12650","article-title":"IL-4 dysregulates microRNAs involved in inflammation, angiogenesis and apoptosis in epidermal keratinocytes","volume":"62","author":"L Bao","year":"2018","journal-title":"Microbio Immunol"},{"issue":"7","key":"pone.0260188.ref049","doi-asserted-by":"crossref","first-page":"e1008622","DOI":"10.1371\/journal.ppat.1008622","article-title":"TLR2 and endosomal TLR-mediated secretion of IL-10 and immune suppression in response to phagosome-confined Listeria monocytogenes","volume":"16","author":"BN Nguyen","journal-title":"PLoS pathog"},{"issue":"14","key":"pone.0260188.ref050","doi-asserted-by":"crossref","first-page":"e103454","DOI":"10.15252\/embj.2019103454","article-title":"Tissue-resident macrophages actively suppress IL-1beta release via a reactive prostanoid\/IL-10 pathway","volume":"39","author":"N Ipseiz","year":"2020","journal-title":"EMBO J"},{"issue":"12","key":"pone.0260188.ref051","doi-asserted-by":"crossref","first-page":"e1004516","DOI":"10.1371\/journal.ppat.1004516","article-title":"P47phox-\/- mice are compromised in expansion and activation of CD8+ T cells and susceptible to Trypanosoma cruzi infection","volume":"10","author":"M Dhiman","year":"2014","journal-title":"PLoS pathog"},{"issue":"10","key":"pone.0260188.ref052","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1111\/j.1365-2362.2011.02499.x","article-title":"Bacteria and spontaneous experimental colitis: immunological changes","volume":"41","author":"E Pedrosa","year":"2011","journal-title":"Eur J Clin Invest"},{"issue":"5","key":"pone.0260188.ref053","first-page":"399","article-title":"Research on the negative immune regulation of NK cells in patients with primary immune thrombocytopenia","volume":"38","author":"YJ Zhang","year":"2017","journal-title":"Zhonghua xue ye xue za zhi"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0260188","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,15]],"date-time":"2023-01-15T20:08:08Z","timestamp":1673813288000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0260188"}},"subtitle":[],"editor":[{"given":"Lili","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2021,11,18]]},"references-count":53,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11,18]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0260188","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,18]]}}}