{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T20:56:00Z","timestamp":1791665760824,"version":"4.3.4"},"reference-count":56,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,2,13]],"date-time":"2022-02-13T00:00:00Z","timestamp":1644710400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Kaken Pharmaceutical (Japan)","award":["No. 19K10325"],"award-info":[{"award-number":["No. 19K10325"]}]},{"name":"Kaken Pharmaceutical (Japan)","award":["No. 21K17194"],"award-info":[{"award-number":["No. 21K17194"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Rupture of the basement membrane in fused palate tissue can cause the palate to separate after fusion in mice, leading to the development of cleft palate. Here, we further elucidate the mechanism of palatal separation after palatal fusion in 8\u201310-week-old ICR female mice. On day 12 of gestation, 40 \u03bcg\/kg of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), sufficient to cause cleft palate in 100% of mice, was dissolved in 0.4 mL of olive oil containing toluene and administered as a single dose via a gastric tube. Fetal palatine frontal sections were observed by H&amp;E staining, and epithelial cell adhesion factors, apoptosis, and cell proliferation were observed from the anterior to posterior palate. TUNEL-positive cells and Ki67-positive cells were observed around the posterior palatal dissection area of the TCDD-treated group. Moreover, in fetal mice exposed to TCDD, some fetuses exhibited cleft palate dehiscence during fusion. The results suggest that palatal dehiscence may be caused by abnormal cell proliferation in epithelial tissues, decreased intercellular adhesion, and inhibition of mesenchymal cell proliferation. By elucidating the mechanism of cleavage after palatal fusion, this research can contribute to establishing methods for the prevention of cleft palate development.<\/jats:p>","DOI":"10.3390\/ijms23042069","type":"journal-article","created":{"date-parts":[[2022,2,13]],"date-time":"2022-02-13T21:08:43Z","timestamp":1644786523000},"page":"2069","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Histological and Immunohistochemical Studies to Determine the Mechanism of Cleft Palate Induction after Palatal Fusion in Mice Exposed to TCDD"],"prefix":"10.3390","volume":"23","author":[{"given":"Chisato","family":"Sakuma","sequence":"first","affiliation":[{"name":"Division of Research and Treatment for Oral and Maxillofacial Congenital Anomalies, School of Dentistry, Aichi-Gakuin University, 2-11 Suemori-Dori, Chikusa-ku, Nagoya 464-8651, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideto","family":"Imura","sequence":"additional","affiliation":[{"name":"Division of Research and Treatment for Oral and Maxillofacial Congenital Anomalies, School of Dentistry, Aichi-Gakuin University, 2-11 Suemori-Dori, Chikusa-ku, Nagoya 464-8651, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohiro","family":"Yamada","sequence":"additional","affiliation":[{"name":"Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Science, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3928-5018","authenticated-orcid":false,"given":"Azumi","family":"Hirata","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Faculty of Medicine, Osaka Medical College, 2-7 Daigaku-Machi, Takatsuki 569-8686, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yayoi","family":"Ikeda","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Dentistry, Aichi-Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaaki","family":"Ito","sequence":"additional","affiliation":[{"name":"Division of Research and Treatment for Oral and Maxillofacial Congenital Anomalies, School of Dentistry, Aichi-Gakuin University, 2-11 Suemori-Dori, Chikusa-ku, Nagoya 464-8651, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nagato","family":"Natsume","sequence":"additional","affiliation":[{"name":"Division of Research and Treatment for Oral and Maxillofacial Congenital Anomalies, School of Dentistry, Aichi-Gakuin University, 2-11 Suemori-Dori, Chikusa-ku, Nagoya 464-8651, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"262e","DOI":"10.1097\/PRS.0000000000008141","article-title":"Update in Unilateral Cleft Lip Surgery","volume":"148","author":"Xue","year":"2021","journal-title":"Plast. Reconstr. Surg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1016\/j.ajoms.2017.03.005","article-title":"A case of Tetrasomy 15q with left cleft lip and alveolus","volume":"29","author":"Imura","year":"2017","journal-title":"J. Oral Maxillofac. Surgery, Med. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/cga.12444","article-title":"Changes in the birth prevalence of orofacial clefts in Japan, Has the birth prevalence of orofacial clefts been affected by improved accuracy of prenatal diagnosis?","volume":"62","author":"Natsume","year":"2022","journal-title":"Congenit. Anom."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"112310","DOI":"10.1016\/j.yexcr.2020.112310","article-title":"The cleft palate candidate gene BAG6 supports FoxO1 acetylation to promote FasL-mediated apoptosis during palate fusion","volume":"396","author":"Xu","year":"2020","journal-title":"Exp. Cell Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1242\/dev.067082","article-title":"Palatogenesis: Morphogenetic and molecular mechanisms of secondary palate development","volume":"139","author":"Bush","year":"2012","journal-title":"Development"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1789","DOI":"10.1242\/dev.02837","article-title":"Multiple functions of Snail family genes during palate development in mice","volume":"134","author":"Murray","year":"2007","journal-title":"Development"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1002\/jmor.1051090204","article-title":"Developmental of cleft palate in mice homozygous for the shorthead mutation","volume":"109","author":"Fitch","year":"1961","journal-title":"J. Morph."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1536","DOI":"10.1002\/dvdy.24177","article-title":"Deciphering TGF-\u03b23 function in medial edge epithelium specification and fusion during mouse secondary palate development","volume":"243","author":"Jin","year":"2014","journal-title":"Dev. Dyn."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1177\/00220345660450032301","article-title":"Mechanisms of fetal posture and casual mechanisms of congenital deformity of the palate, mandible and limbs","volume":"45","author":"Pratt","year":"1966","journal-title":"J. Dent. Res."},{"key":"ref_10","first-page":"240","article-title":"Epithelial remnants and pearls in the secondary palate in the human abortus: A contribution to the study of the mechanism of cleft palate formation","volume":"3","author":"Kitamura","year":"1966","journal-title":"Cleft Palate J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1597\/1545-1569_1991_028_0195_efcpaa_2.3.co_2","article-title":"Evidence for cleft palate as a postfusion phenomenon","volume":"28","author":"Kitamura","year":"1990","journal-title":"Cleft Palate J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.bjoms.2014.12.008","article-title":"Association of single-nucleotide polymorphisms in the IRF6 gene with non-syndromic cleft lip with or without cleft palate in the Xinjiang Uyghur population","volume":"53","author":"Mijiti","year":"2015","journal-title":"Br. J. Oral Maxillofac. Surg."},{"key":"ref_13","first-page":"9","article-title":"Genetic analysis of MEOX2 in Japanese non-syndromic cleft palate patients","volume":"29","author":"Mori","year":"2016","journal-title":"Aichi Gakuin Dent. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1111\/cga.12259","article-title":"Association of MEOX2 polymorphism with nonsyndromic cleft palate only in a Vietnamese population","volume":"58","author":"Tran","year":"2017","journal-title":"Congenit. Anomalies"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1111\/cga.12153","article-title":"Homeobox family Hoxc localization during murine palate formation","volume":"56","author":"Hirata","year":"2016","journal-title":"Congenit. Anomalies"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Vaivads, M., Akota, I., and Pilmane, M. (2021). Cleft Candidate Genes and Their Products in Human Unilateral Cleft Lip Tissue. Diseases, 9.","DOI":"10.3390\/diseases9020026"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e1754","DOI":"10.1002\/mgg3.1754","article-title":"Family based and case-control designs reveal an association of TFAP2A in non-syndromic cleft lip only among Vietnamese population","volume":"9","author":"Nguyen","year":"2021","journal-title":"Mol. Genet. Genomic Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1016\/j.bjoms.2015.05.017","article-title":"Maternal obesity is a risk factor for orofacial clefts: A meta-analysis","volume":"53","author":"Blanco","year":"2015","journal-title":"Br. J. Oral Maxillofac. Surg."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Jaruga, A., Ksiazkiewicz, J., Kuzniarz, K., and Tylzanowski, P. (2022). Orofacial Cleft and Mandibular Prognathism\u2014Human Genetics and Animal Models. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23020953"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"103270","DOI":"10.1016\/j.etap.2019.103270","article-title":"Common differentially expressed proteins were found in mouse cleft palate models induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin and retinoic acid","volume":"72","author":"Wang","year":"2019","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/0272-0590(90)90171-F","article-title":"Characterization of the period of sensitivity for the induction of hydronephrosis in C57BL\/6N mice following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin","volume":"15","author":"Couture","year":"1990","journal-title":"Fundam. Appl. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1111\/j.1741-4520.2006.00097.x","article-title":"Cleft lip and palate in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin: A morphological in vivo study","volume":"46","author":"Yamada","year":"2006","journal-title":"Congenit. Anomalies"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1111\/j.1741-4520.2010.00271.x","article-title":"Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin suggests abnormal palate development after palatal fusion","volume":"50","author":"Imura","year":"2010","journal-title":"Congenit. Anomalies"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2027","DOI":"10.1016\/j.jcms.2018.09.016","article-title":"Cleft palate formation after palatal fusion occurs due to the rupture of epithelial basement membranes","volume":"46","author":"Sakuma","year":"2018","journal-title":"J. Cranio-Maxillofacial Surg."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1093\/toxsci\/66.1.117","article-title":"Effect of a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin on immune function in male NC\/Nga mice","volume":"66","author":"Fujimaki","year":"2002","journal-title":"Toxicol. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.toxlet.2020.08.001","article-title":"Comparison of the biological behaviors of palatal mesenchymal and epithelial cells induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in vitro","volume":"333","author":"Chen","year":"2020","journal-title":"Toxicol. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1111\/1523-1747.ep12456367","article-title":"Effect of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin on Murine Skin","volume":"90","author":"Puhvel","year":"1988","journal-title":"J. Investig. Dermatol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"340","DOI":"10.3389\/fcell.2019.00340","article-title":"Extracellular Matrix Composition and Remodeling: Current Perspectives on Secondary Palate Formation, Cleft Lip\/Palate, and Palatal Reconstruction","volume":"7","author":"Paiva","year":"2019","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1007\/s12253-019-00614-3","article-title":"MEG3: An Oncogenic Long Non-coding RNA in Different Cancers","volume":"25","author":"Alanazi","year":"2019","journal-title":"Pathol. Oncol. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"115517","DOI":"10.1016\/j.taap.2021.115517","article-title":"The role of MEG3 in the proliferation of palatal mesenchymal cells is related to the TGF\u03b2\/Smad pathway in TCDD inducing cleft palate","volume":"419","author":"He","year":"2021","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1046\/j.1365-2443.1997.1490345.x","article-title":"Loss of teratogenic response to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in mice lacking the Ah (dioxin) receptor","volume":"2","author":"Mimura","year":"1997","journal-title":"Genes Cells"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Yoshioka, W., and Tohyama, C. (2019). Mechanisms of Developmental Toxicity of Dioxins and Related Compounds. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20030617"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"152353","DOI":"10.1016\/j.tox.2019.152353","article-title":"TCDD inhibited the osteogenic differentiation of human fetal palatal mesenchymal cells through AhR and BMP-2\/TGF-\u03b2\/Smad signaling","volume":"15","author":"Liu","year":"2020","journal-title":"Toxicology."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1002\/(SICI)1520-6866(2000)20:2<73::AID-TCM3>3.0.CO;2-M","article-title":"Pathogenesis of cleft palate in mouse embryos exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)","volume":"20","author":"Takagi","year":"2000","journal-title":"Teratog. Carcinog. Mutagen."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1021\/acs.chemrestox.6b00350","article-title":"Computational Model of Secondary Palate Fusion and Disruption","volume":"30","author":"Hutson","year":"2017","journal-title":"Chem Res. Toxicol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1083\/jcb.95.1.333","article-title":"Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells","volume":"95","author":"Greenburg","year":"1982","journal-title":"J. Cell Biol."},{"key":"ref_37","first-page":"71","article-title":"Epithelial to mesenchymal transition in tumor cells as consequence of phenotypic instability","volume":"2","year":"2014","journal-title":"Front. Cell. Dev. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1111\/jop.12382","article-title":"E-cadherin regulators are differentially expressed in the epithelium and stroma of keratocystic odontogenic tumors","volume":"45","author":"Porto","year":"2015","journal-title":"J. Oral Pathol. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/S0012-1606(89)80017-X","article-title":"Medial edge epithelium transforms to mesenchyme after embryonic palatal shelves fuse","volume":"131","author":"Fitchett","year":"1989","journal-title":"Dev. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1111\/j.1440-169X.2010.01245.x","article-title":"Disintegration of the medial epithelial seam: Is cell death important in palatogenesis?","volume":"53","author":"Iseki","year":"2011","journal-title":"Dev. Growth. Differ."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.taap.2016.06.016","article-title":"Effect of TCDD on the fate of epithelial cells isolated from human fetal palatal shelves (hFPECs)","volume":"305","author":"Gao","year":"2016","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_42","first-page":"438","article-title":"Oct4 plays a role in 2,3,7,8-tetrachlorobenzo-p-dioxin (TCDD) inducing cleft palate and inhibiting mesenchymal proliferation","volume":"30","author":"Tao","year":"2020","journal-title":"Toxicology"},{"key":"ref_43","first-page":"140","article-title":"Loss of basement membrane components by invasive tumors but not by their benign counterparts","volume":"49","author":"Barsky","year":"1983","journal-title":"Lab. Investig."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3115","DOI":"10.1158\/0008-5472.CAN-16-1297","article-title":"Adhere, Degrade, and Move: The Three-Step Model of Invasion","volume":"76","author":"Liotta","year":"2016","journal-title":"Cancer Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1242\/jcs.113.5.859","article-title":"Assembly of the exogenous extracellular matrix during basement membrane formation by alveolar epithelial cells in vitro","volume":"113","author":"Furuyama","year":"2000","journal-title":"J. Cell Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1080\/15476278.2020.1735239","article-title":"Extracellular Matrix Remodeling During Palate Development","volume":"16","author":"Wang","year":"2020","journal-title":"Organogenesis"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/978-94-007-7893-1_3","article-title":"Basic Components of Connective Tissues and Extracellular Matrix: Elastin, Fibrillin, Fibulins, Fibrinogen, Fibronectin, Laminin, Tenascins and Thrombospondins","volume":"802","author":"Halper","year":"2013","journal-title":"Prog. Heritable Soft Connect. Tissue Dis."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1111\/j.1469-7580.2006.00549.x","article-title":"Extracellular matrix adaptation of tendon and skeletal muscle to exercise","volume":"208","author":"Kjaer","year":"2006","journal-title":"J. Anat."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1002\/tera.1420350307","article-title":"TCDD-Induced hyperplasia of the ureteral epithelium produces hydronephrosis in murine fetuses","volume":"35","author":"Abbott","year":"1987","journal-title":"Teratology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/0041-008X(89)90010-0","article-title":"TCDD alters medial epithelial cell differentiation during palatogenesis","volume":"99","author":"Abbott","year":"1989","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0300-483X(01)00362-6","article-title":"Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on fetal mouse urinary tract epithelium in vitro","volume":"162","author":"Bryant","year":"2001","journal-title":"Toxicology"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1002\/lio2.214","article-title":"Cellular and molecular mechanisms of cleft palate development","volume":"4","author":"Deshpande","year":"2018","journal-title":"Laryngoscope"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1007\/s00441-014-1994-6","article-title":"Defects in the adherens junction complex (E-cadherin\/\u03b2-catenin) in inflammatory bowel disease","volume":"360","author":"Mehta","year":"2015","journal-title":"Cell Tissue Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1126\/science.2006419","article-title":"Cadherin Cell Adhesion Receptors as a Morphogenetic Regulator","volume":"251","author":"Takeichi","year":"1991","journal-title":"Science"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1038\/nrc822","article-title":"Epithelial-mesenchymal transitions in tumor progression","volume":"2","author":"Theiery","year":"2002","journal-title":"Nat. Rev. Cancer"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1111\/j.1741-4520.2008.00181.x","article-title":"Morphological and immunohistochemical studies on cleft palates induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice","volume":"48","author":"Fujiwara","year":"2008","journal-title":"Congenit. Anom."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/4\/2069\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:18:50Z","timestamp":1760134730000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/4\/2069"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,13]]},"references-count":56,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23042069"],"URL":"https:\/\/doi.org\/10.3390\/ijms23042069","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,13]]}}}