{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T05:57:25Z","timestamp":1790834245699,"version":"4.1.0"},"reference-count":56,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2019,4,11]],"date-time":"2019-04-11T00:00:00Z","timestamp":1554940800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["AGL2015-69738-R"],"award-info":[{"award-number":["AGL2015-69738-R"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","award":["AGL2015-69738-R"],"award-info":[{"award-number":["AGL2015-69738-R"]}],"id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007801","name":"Fundaci\u00f3n S\u00e9neca","doi-asserted-by":"publisher","award":["19892\/GERM-15"],"award-info":[{"award-number":["19892\/GERM-15"]}],"id":[{"id":"10.13039\/100007801","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001862","name":"Svenska Forskningsr\u00e5det Formas","doi-asserted-by":"publisher","award":["2017-00946"],"award-info":[{"award-number":["2017-00946"]}],"id":[{"id":"10.13039\/501100001862","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010805","name":"Forskningsr\u00e5det i Syd\u00f6stra Sverige","doi-asserted-by":"publisher","award":["745971"],"award-info":[{"award-number":["745971"]}],"id":[{"id":"10.13039\/100010805","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS\/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein\u2013protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.<\/jats:p>","DOI":"10.3390\/ijms20071791","type":"journal-article","created":{"date-parts":[[2019,4,12]],"date-time":"2019-04-12T03:46:37Z","timestamp":1555040797000},"page":"1791","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":47,"title":["Cryopreservation Differentially Alters the Proteome of Epididymal and Ejaculated Pig Spermatozoa"],"prefix":"10.3390","volume":"20","author":[{"given":"Cristina","family":"Perez-Pati\u00f1o","sequence":"first","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9873-814X","authenticated-orcid":false,"given":"Isabel","family":"Barranco","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"},{"name":"Department of Biology, Faculty of Sciences, University of Girona, 17003 Girona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junwei","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"},{"name":"School of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorena","family":"Padilla","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilio A.","family":"Martinez","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5194-2124","authenticated-orcid":false,"given":"Heriberto","family":"Rodriguez-Martinez","sequence":"additional","affiliation":[{"name":"Department of Clinical and Experimental Medicine, Faculty of Medicine and Health Sciences, Link\u00f6ping University, SE-58185 Link\u00f6ping, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4213-3093","authenticated-orcid":false,"given":"Jordi","family":"Roca","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inmaculada","family":"Parrilla","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,4,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/s11934-018-0803-2","article-title":"Male Fertility Preservation","volume":"19","author":"Lipshultz","year":"2018","journal-title":"Curr. Urol. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Kawai, K., and Nishiyama, H. (2018). Preservation of fertility of adult male cancer patients treated with chemotherapy. Int. J. Clin. Oncol., in press.","DOI":"10.1007\/s10147-018-1333-0"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1017\/S0967199417000442","article-title":"Reproduction biotechnologies in germplasm banking of livestock species: A review","volume":"25","author":"Morrell","year":"2017","journal-title":"Zygote"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"640","DOI":"10.4103\/1008-682X.153849","article-title":"Biobanking efforts and new advances in male fertility preservation for rare and endangered species","volume":"17","author":"Comizzoli","year":"2015","journal-title":"Asian J. Androl."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.cveq.2016.08.003","article-title":"Advances in Stallion Semen Cryopreservation","volume":"32","author":"Alvarenga","year":"2016","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1002\/mrd.22840","article-title":"Artificial insemination with frozen-thawed boar sperm","volume":"84","author":"Yeste","year":"2017","journal-title":"Mol. Reprod. Dev."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Rozati, H., Handley, T., and Jayasena, C.N. (2017). Process and Pitfalls of Sperm Cryopreservation. J. Clin. Med., 6.","DOI":"10.3390\/jcm6090089"},{"key":"ref_8","first-page":"115","article-title":"To What Extent Are Cryopreserved Sperm and Testicular Biopsy Samples Used in Assisted Reproduction?","volume":"19","author":"Rosenbusch","year":"2018","journal-title":"J. Reprod. Infertil."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3109\/19396368.2014.976720","article-title":"Effect of sperm storage and selection techniques on sperm parameters","volume":"61","author":"Sharma","year":"2015","journal-title":"Syst. Biol. Reprod. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1111\/rda.12552","article-title":"The Fertility of Frozen Boar Sperm When used for Artificial Insemination","volume":"50","author":"Knox","year":"2015","journal-title":"Reprod. Domest. Anim."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.theriogenology.2018.06.006","article-title":"Sublethal sperm freezing damage: Manifestations and solutions","volume":"118","author":"Pini","year":"2018","journal-title":"Theriogenology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.cryobiol.2011.12.002","article-title":"Freezing injury: The special case of the sperm cell","volume":"64","author":"Morris","year":"2012","journal-title":"Cryobiology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.anireprosci.2016.08.008","article-title":"Sperm proteome and reproductive technologies in mammals","volume":"173","author":"Li","year":"2016","journal-title":"Anim. Reprod. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.rbmo.2018.05.012","article-title":"Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches","volume":"37","author":"Hezavehei","year":"2018","journal-title":"Reprod. Biomed. Online"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1016\/S0093-691X(99)00046-1","article-title":"Substantial decrease of heat-shock protein 90 precedes the decline of sperm motility during cooling of boar spermatozoa","volume":"51","author":"Huang","year":"1999","journal-title":"Theriogenology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1002\/j.1939-4640.2000.tb02138.x","article-title":"Cryopreservation alters the levels of the bull sperm surface protein P25b","volume":"21","author":"Lessard","year":"2000","journal-title":"J. Androl."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1002\/mrd.1115","article-title":"Cryopreservation alters membrane sulfhydryl status of bull spermatozoa: Protection by oxidized glutathione","volume":"60","author":"Chatterjee","year":"2001","journal-title":"Mol. Reprod. Dev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.theriogenology.2010.04.021","article-title":"The HSP90AA1 sperm content and the prediction of the boar ejaculate freezability","volume":"74","author":"Casas","year":"2010","journal-title":"Theriogenology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.theriogenology.2010.02.018","article-title":"Cryopreservation-induced alterations in boar spermatozoa mitochondrial function are related to changes in the expression and location of midpiece mitofusin-2 and actin network","volume":"74","author":"Flores","year":"2010","journal-title":"Theriogenology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1002\/pmic.201300225","article-title":"Proteomic characteristics of human sperm cryopreservation","volume":"14","author":"Wang","year":"2014","journal-title":"Proteomics"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1111\/andr.12279","article-title":"Identification of protein changes in human spermatozoa throughout the cryopreservation process","volume":"5","author":"Bogle","year":"2017","journal-title":"Andrology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.cryobiol.2015.11.003","article-title":"Differential proteome association study of freeze-thaw damage in ram sperm","volume":"72","author":"He","year":"2016","journal-title":"Cryobiology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.jprot.2018.04.001","article-title":"Cryopreservation and egg yolk medium alter the proteome of ram spermatozoa","volume":"181","author":"Pini","year":"2018","journal-title":"J. Proteom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1530\/REP-13-0313","article-title":"Identification of differentially expressed proteins in fresh and frozenethawed boar spermatozoa by iTRAQcoupled 2D LCeMS\/MS","volume":"147","author":"Chen","year":"2014","journal-title":"Reproduction"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Rothschild, M.F., and Ruvinsky, A. (2011). Pigs as a model of biochemical science. The Genetics of the Pig, CAB International. [2nd ed.].","DOI":"10.1079\/9781845937560.0000"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1016\/j.fertnstert.2013.05.020","article-title":"Dead spermatozoa in raw semen samples impair in vitro fertilization outcomes of frozen-thawed spermatozoa","volume":"100","author":"Roca","year":"2013","journal-title":"Fertil. Steril."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.cryobiol.2014.07.004","article-title":"Boar sperm cryosurvival is better after exposure to seminal plasma from selected fractions than to those from entire ejaculate","volume":"69","author":"Alkmin","year":"2014","journal-title":"Cryobiology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1016\/S0093-691X(97)00034-4","article-title":"In vitro fertilization of porcine oocytes with fresh and frozen-thawed ejaculated or frozen-thawed epididymal semen obtained from identical boars","volume":"47","author":"Rath","year":"1997","journal-title":"Theriogenology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1016\/j.theriogenology.2004.09.018","article-title":"The significance of cooling rates and animal variability for boar sperm cryopreservation: Insights from the cryomicroscope","volume":"63","author":"Holt","year":"2005","journal-title":"Theriogenology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2692","DOI":"10.2527\/jas.2006-094","article-title":"Factors influencing boar sperm cryosurvival","volume":"84","author":"Roca","year":"2006","journal-title":"J. Anim. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.theriogenology.2015.09.047","article-title":"Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs","volume":"85","author":"Yeste","year":"2016","journal-title":"Theriogenology"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.theriogenology.2013.05.006","article-title":"Acrosin-binding protein (ACRBP) and triosephosphate isomerase (TPI) are good markers to predict boar sperm freezing capacity","volume":"80","author":"Vilagran","year":"2013","journal-title":"Theriogenology"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/j.theriogenology.2014.04.023","article-title":"Relationship of sperm small heat-shock protein 10 and voltage-dependent anion channel 2 with semen freezability in boars","volume":"82","author":"Vilagran","year":"2014","journal-title":"Theriogenology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.anireprosci.2017.06.005","article-title":"Sperm membrane proteins associated with the boar semen cryopreservation","volume":"183","author":"Barros","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1074\/mcp.RA118.000840","article-title":"The proteome of pig spermatozoa is remodeled during ejaculation","volume":"18","author":"Parrilla","year":"2019","journal-title":"Mol. Cell. Proteom."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/j.1600-0897.2011.01033.x","article-title":"Seminal plasma proteins: What role do they play?","volume":"66","author":"Kvist","year":"2011","journal-title":"Am. J. Reprod. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3700","DOI":"10.1021\/acs.jproteome.6b00530","article-title":"Proteomic Investigation of Ram Spermatozoa and the Proteins Conferred by Seminal Plasma","volume":"15","author":"Pini","year":"2016","journal-title":"J. Proteome Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.jprot.2016.04.026","article-title":"Characterization of the porcine seminal plasma proteome comparing ejaculate portions","volume":"142","author":"Barranco","year":"2016","journal-title":"J. Proteom."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5240","DOI":"10.2527\/jas.2015-9237","article-title":"Impact of cryopreservation on bull () semen proteome","volume":"93","author":"Westfalewicz","year":"2015","journal-title":"J. Anim. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1186\/s40104-016-0128-2","article-title":"Proteomic identification of cryostress in epididymal spermatozoa","volume":"7","author":"Yoon","year":"2016","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1093\/molehr\/gap077","article-title":"Sperm surface proteomics: From protein lists to biological function","volume":"16","author":"Brewis","year":"2010","journal-title":"Mol. Hum. Reprod."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1111\/j.1365-2605.2007.00851.x","article-title":"The role of proteomics in understanding sperm cell biology","volume":"31","author":"Aitken","year":"2008","journal-title":"Int. J. Androl."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.anireprosci.2018.04.061","article-title":"Seminal plasma proteomes and sperm fertility","volume":"194","author":"Druart","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.theriogenology.2013.10.010","article-title":"Cryogenic changes in proteases and antiprotease activities of buffalo (Bubalus bubalis) and cattle (Bos taurus) semen","volume":"81","author":"Gurupriya","year":"2014","journal-title":"Theriogenology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/bse0470001","article-title":"The mitochondrial respiratory chain","volume":"47","author":"Rich","year":"2010","journal-title":"Essays Biochem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/j.bbrc.2013.05.095","article-title":"DJ-1 cooperates with PYCR1 in cell protection against oxidative stress","volume":"436","author":"Yasuda","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"6136075","DOI":"10.1155\/2018\/6136075","article-title":"Downregulation of DJ-1 Fails to Protect Mitochondrial Complex I Subunit NDUFS3 in the Testes and Contributes to the Asthenozoospermia","volume":"2018","author":"Wang","year":"2018","journal-title":"Mediat. Inflamm."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.anireprosci.2018.03.024","article-title":"The sperm mitochondrion: Organelle of many functions","volume":"194","author":"Moraes","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1071\/RD14088","article-title":"Seminal plasma affects sperm sorting in boars","volume":"28","author":"Alkmin","year":"2016","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1038\/s41598-018-36624-5","article-title":"The proteome of frozen-thawed pig spermatozoa is dependent on the ejaculate fraction source","volume":"9","author":"Li","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"14440","DOI":"10.1073\/pnas.93.25.14440","article-title":"Linking genome and proteome by mass spectrometry: Large-scale identification of yeast proteins from two dimensional gels","volume":"93","author":"Shevchenko","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Gillet, L.C., Navarro, P., Tate, S., R\u00f6st, H., Selevsek, N., Reiter, L., Bonner, R., and Aebersold, R. (2012). Targeted data extraction of the MS\/MS spectra generated by data-independent acquisition: A new concept for consistent and accurate proteome analysis. Mol. Cell. Proteom., 11.","DOI":"10.1074\/mcp.O111.016717"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1038\/nprot.2008.211","article-title":"Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources","volume":"4","author":"Huang","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/nar\/gkn923","article-title":"Bioinformatics enrichment tools: Paths toward the comprehensive functional analysis of large gene lists","volume":"37","author":"Huang","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: A software environment for integrated models of biomolecular interaction networks","volume":"13","author":"Shannon","year":"2003","journal-title":"Genome Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1093\/bioinformatics\/btp101","article-title":"ClueGO: A Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks","volume":"25","author":"Bindea","year":"2009","journal-title":"Bioinformatics"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/7\/1791\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:44:32Z","timestamp":1760186672000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/7\/1791"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,4,11]]},"references-count":56,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2019,4]]}},"alternative-id":["ijms20071791"],"URL":"https:\/\/doi.org\/10.3390\/ijms20071791","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,4,11]]}}}