{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:31:12Z","timestamp":1760059872816,"version":"build-2065373602"},"reference-count":57,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,15]],"date-time":"2025-07-15T00:00:00Z","timestamp":1752537600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute of Food and Agriculture, U.S. Department of Agriculture","award":["#1016077","6066-31000-015-00D","P20GM103476"],"award-info":[{"award-number":["#1016077","6066-31000-015-00D","P20GM103476"]}]},{"name":"USDA-ARS","award":["#1016077","6066-31000-015-00D","P20GM103476"],"award-info":[{"award-number":["#1016077","6066-31000-015-00D","P20GM103476"]}]},{"name":"Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health","award":["#1016077","6066-31000-015-00D","P20GM103476"],"award-info":[{"award-number":["#1016077","6066-31000-015-00D","P20GM103476"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Artificial insemination (AI) is a key breeding technique in the swine industry; however, the lack of reliable biomarkers for semen quality limits its effectiveness. Seminal plasma (SP) contains extracellular vesicles (EVs) that present a promising, non-invasive biomarker for semen quality. This study explores the biochemical profiles of boar SP to assess semen quality through near-infrared spectroscopy (NIRS) and proteomics of SP-EVs. Fresh semen from mature Duroc boars was evaluated based on sperm motility, classifying samples as Passed (\u226570%) or Failed (&lt;70%). NIRS analysis identified distinct variations in water structures at specific wavelengths (C1, C5, C12 nm), achieving high accuracy (92.2%), sensitivity (94.2%), and specificity (90.3%) through PCA-LDA. Proteomic analysis of SP-EVs revealed 218 proteins in Passed and 238 in Failed samples. Nexin-1 and seminal plasma protein pB1 were upregulated in Passed samples, while LGALS3BP was downregulated. The functional analysis highlighted pathways associated with single fertilization, filament organization, and glutathione metabolism in Passed samples. Integrating NIRS with SP-EV proteomics provides a robust approach to non-invasive assessment of semen quality. These findings suggest that SP-EVs could serve as effective biosensors for rapid semen quality assessment, enabling better boar semen selection and enhancing AI practices in swine breeding.<\/jats:p>","DOI":"10.3390\/bios15070456","type":"journal-article","created":{"date-parts":[[2025,7,15]],"date-time":"2025-07-15T15:01:05Z","timestamp":1752591665000},"page":"456","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrating Near-Infrared Spectroscopy and Proteomics for Semen Quality Biosensing"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3644-6576","authenticated-orcid":false,"given":"Notsile H.","family":"Dlamini","sequence":"first","affiliation":[{"name":"Department of Animal and Dairy Sciences, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3356-6276","authenticated-orcid":false,"given":"Mariana","family":"Santos-Rivera","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Nutrition, and Health Promotion, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8466-6077","authenticated-orcid":false,"given":"Carrie K.","family":"Vance-Kouba","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Nutrition, and Health Promotion, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olga","family":"Pechanova","sequence":"additional","affiliation":[{"name":"Institute of Genomics, Biocomputing, and Bioinformatics, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tibor","family":"Pechan","sequence":"additional","affiliation":[{"name":"Institute of Genomics, Biocomputing, and Bioinformatics, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1376-5059","authenticated-orcid":false,"given":"Jean M.","family":"Feugang","sequence":"additional","affiliation":[{"name":"Department of Animal and Dairy Sciences, Mississippi State University, Starkville, MS 39762, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1016\/j.theriogenology.2008.06.018","article-title":"Diseases in swine transmitted by artificial insemination: An overview","volume":"70","author":"Maes","year":"2008","journal-title":"Theriogenology"},{"key":"ref_2","first-page":"67","article-title":"Selection for boar fertility and semen quality\u2014The way ahead","volume":"66","author":"Flowers","year":"2009","journal-title":"Soc. Reprod. Fertil."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Mann, T., and Lutwak-Mann, C. (1981). Male reproductive function and the composition of semen: General considerations. Male Reproductive Function and Semen, Springer.","DOI":"10.1007\/978-1-4471-1300-3"},{"key":"ref_4","first-page":"13","article-title":"Seminal plasma effects on sperm handling and female fertility","volume":"64","author":"Maxwell","year":"2007","journal-title":"Soc. Reprod. Fertil. Suppl."},{"key":"ref_5","first-page":"1234","article-title":"The epididymis and its role on sperm quality and male fertility","volume":"14","author":"Lucca","year":"2018","journal-title":"Anim. Reprod. (AR)"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.theriogenology.2023.09.026","article-title":"The effects of boar seminal plasma extracellular vesicles on sperm fertility","volume":"213","author":"Xu","year":"2024","journal-title":"Theriogenology"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Dlamini, N.H., Nguyen, T., Gad, A., Tesfaye, D., Liao, S.F., Willard, S.T., Ryan, P.L., and Feugang, J.M. (2023). Characterization of Extracellular Vesicle-Coupled miRNA Profiles in Seminal Plasma of Boars with Divergent Semen Quality Status. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24043194"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1111\/rda.14453","article-title":"Metabolomic profile of seminal plasma from Guzer\u00e1 bulls (Bos indicus) with contrasting sperm freezability phenotypes","volume":"58","author":"Pessoa","year":"2023","journal-title":"Reprod. Domest. Anim."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107051","DOI":"10.1016\/j.anireprosci.2022.107051","article-title":"Extracellular vesicles in seminal plasma: A safe and relevant tool to improve fertility in livestock?","volume":"244","author":"Roca","year":"2022","journal-title":"Anim. Reprod. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"\u00c1lvarez-Rodr\u00edguez, M., and Martinez-Pastor, F. (2021). Martinez-Pastor, Molecular determinants of seminal plasma on sperm biology and fertility. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073555"},{"key":"ref_11","unstructured":"Shaw, R.A., and Mantsch, H.H. (2006). Infrared spectroscopy of biological fluids in clinical and diagnostic analysis. Encyclopedia of Analytical Chemistry: Applications, Theory and Instrumentation, Wiley Online Library."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1590\/S0103-50532003000200006","article-title":"Near infrared spectroscopy: Fundamentals, practical aspects and analytical applications","volume":"14","author":"Pasquini","year":"2003","journal-title":"J. Braz. Chem. Soc."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Williams, P., Manley, M., and Antoniszyn, J. (2019). Near Infrared Technology: Getting the Best out of Light, African Sun Media.","DOI":"10.18820\/9781928480310"},{"key":"ref_14","first-page":"14","article-title":"AquaPhotomics: Water absorbance pattern as a biological marker for disease diagnosis and disease understanding","volume":"18","author":"Tsenkova","year":"2007","journal-title":"NIR News"},{"key":"ref_15","unstructured":"Sutovsky, P., and Lovercamp, K. (2011). Molecular markers of sperm quality. Reproduction in Domestic Ruminants VII: Proceedings of the 8th International Symposium on Reproduction in Domestic Ruminants, Anchorage, Alaska, September 2021, Nottingham University Press."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Candenas, L., and Chianese, R. (2020). Exosome composition and seminal plasma proteome: A promising source of biomarkers of male infertility. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21197022"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1083\/jcb.201211138","article-title":"Extracellular vesicles: Exosomes, microvesicles, and friends","volume":"200","author":"Raposo","year":"2013","journal-title":"J. Cell Biol."},{"key":"ref_18","first-page":"17","article-title":"Lectin-functionalized magnetic iron oxide nanoparticles for reproductive improvement","volume":"3","author":"Feugang","year":"2015","journal-title":"JFIV Reprod. Med. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Gebremedhn, S., Gad, A., Aglan, H.S., Laurincik, J., Prochazka, R., Salilew-Wondim, D., Hoelker, M., Schellander, K., and Tesfaye, D. (2020). Extracellular vesicles shuttle protective messages against heat stress in bovine granulosa cells. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-72706-z"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Feugang, J.M., Liao, S.F., Willard, S.T., and Ryan, P.L. (2018). In-depth proteomic analysis of boar spermatozoa through shotgun and gel-based methods. BMC Genom., 19.","DOI":"10.1186\/s12864-018-4442-2"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Feugang, J.M., Rodriguez-Munoz, J.C., Willard, S.T., Bathgate, R.A., and Ryan, P.L. (2011). Examination of relaxin and its receptors expression in pig gametes and embryos. Reprod. Biol. Endocrinol., 9.","DOI":"10.1186\/1477-7827-9-10"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Feugang, J.M., Greene, J.M., Sanchez-Rodr\u00edguez, H.L., Stokes, J.V., Crenshaw, M.A., Willard, S.T., and Ryan, P.L. (2015). Profiling of relaxin and its receptor proteins in boar reproductive tissues and spermatozoa. Reprod. Biol. Endocrinol., 13.","DOI":"10.1186\/s12958-015-0043-y"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1016\/S0093-691X(00)00261-2","article-title":"Field investigations of bacterial contaminants and their effects on extended porcine semen","volume":"53","author":"Althouse","year":"2000","journal-title":"Theriogenology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1016\/S0093-691X(03)00248-6","article-title":"Regression analyses and motile sperm subpopulation structure study as improving tools in boar semen quality analysis","volume":"61","author":"Rigau","year":"2004","journal-title":"Theriogenology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1751-0147-54-1","article-title":"Impact of proximal cytoplasmic droplets on quality traits and in-vitro embryo production efficiency of cryopreserved bull spermatozoa","volume":"54","author":"Carreira","year":"2012","journal-title":"Acta Veter. Scand."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1023\/A:1009420432234","article-title":"Andrology: Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro","volume":"18","author":"Hirano","year":"2001","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s40813-017-0062-5","article-title":"Boar management and semen handling factors affect the quality of boar extended semen","volume":"3","author":"Rodriguez","year":"2017","journal-title":"Porc. Health Manag."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Tsenkova, R., Mun\u0107an, J., Pollner, B., and Kovacs, Z. (2018). Essentials of aquaphotomics and its chemometrics approaches. Front. Chem., 6.","DOI":"10.3389\/fchem.2018.00363"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Muncan, J., and Tsenkova, R. (2019). Aquaphotomics\u2014From Innovative Knowledge to Integrative Platform in Science and Technology. Molecules, 24.","DOI":"10.3390\/molecules24152742"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"105734","DOI":"10.1016\/j.compag.2020.105734","article-title":"Online milk composition analysis with an on-farm near-infrared sensor","volume":"178","author":"Adriaens","year":"2020","journal-title":"Comput. Electron. Agric."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1111\/j.1745-7262.2007.00281.x","article-title":"Epididymosomes are involved in the acquisition of new sperm proteins during epididymal transit","volume":"9","author":"Sullivan","year":"2007","journal-title":"Asian J. Androl."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7290","DOI":"10.1093\/nar\/gku347","article-title":"Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions","volume":"42","author":"Vojtech","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-019-48095-3","article-title":"Extracellular vesicles isolated from porcine seminal plasma exhibit different tetraspanin expression profiles","volume":"9","author":"Barranco","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"58832","DOI":"10.18632\/oncotarget.11315","article-title":"Boar seminal plasma exosomes maintain sperm function by infiltrating into the sperm membrane","volume":"7","author":"Du","year":"2016","journal-title":"Oncotarget"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Rowlison, T., and Comizzoli, P. (2023). Transfer of Galectin-3-Binding Protein via Epididymal Extracellular Vesicles Promotes Sperm Fertilizing Ability and Developmental Potential in the Domestic Cat Model. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24043077"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Xu, Z., Xie, Y., Zhou, C., Hu, Q., Gu, T., Yang, J., Zheng, E., Huang, S., Xu, Z., and Cai, G. (2020). Expression pattern of seminal plasma extracellular vesicle small RNAs in boar semen. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.585276"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Al-Dossary, A.A., Strehler, E.E., Martin-DeLeon, P.A., and Drevet, J.R. (2013). Expression and secretion of plasma membrane Ca2+-ATPase 4a (PMCA4a) during murine estrus: Association with oviductal exosomes and uptake in sperm. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0080181"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.theriogenology.2020.05.043","article-title":"Porcine oviductal extracellular vesicles interact with gametes and regulate sperm motility and survival","volume":"155","author":"Schmaltz","year":"2020","journal-title":"Theriogenology"},{"key":"ref_39","first-page":"106853","article-title":"Extracellular vesicles in seminal fluid and effects on male reproduction. Overv. Farm Anim. Pets","volume":"16","author":"Roca","year":"2021","journal-title":"Anim. Reprod. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/bs.acc.2019.08.004","article-title":"Exosomes of male reproduction","volume":"95","author":"Baskaran","year":"2020","journal-title":"Adv. Clin. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Wang, D., Jueraitetibaike, K., Tang, T., Wang, Y., Jing, J., Xue, T., Ma, J., Cao, S., Lin, Y., and Li, X. (2021). Seminal plasma and seminal plasma exosomes of aged male mice affect early embryo implantation via immunomodulation. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.723409"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1017\/S0967199422000090","article-title":"Extracellular vesicles in mammalian reproduction: A review","volume":"30","author":"Fuentes","year":"2022","journal-title":"Zygote"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1477-7827-6-5","article-title":"Prostasome-like vesicles stimulate acrosome reaction of pig spermatozoa","volume":"6","author":"Siciliano","year":"2008","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1515\/bchm3.1995.376.4.237","article-title":"T\u00f6pfer-Petersen, Interaction of non-aggregated boar AWN-1 and AQN-3 with phospholipid matrices. A model for coating of spermadhesins to the sperm surface","volume":"376","author":"Calvete","year":"1995","journal-title":"Biol. Chem. Hoppe-Seyler"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.1095\/biolreprod.104.036426","article-title":"Identification, proteomic profiling, and origin of ram epididymal fluid exosome-like vesicles","volume":"72","author":"Gatti","year":"2005","journal-title":"Biol. Reprod."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1074\/mcp.RA119.001594","article-title":"Proteomics profiling of autologous blood and semen exosomes from HIV-infected and uninfected individuals reveals compositional and functional variabilities","volume":"19","author":"Kaddour","year":"2020","journal-title":"Mol. Cell. Proteom."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Llavanera, M., Delgado-Berm\u00fadez, A., Olives, S., Mateo-Otero, Y., Recuero, S., Bonet, S., Fern\u00e1ndez-Fuertes, B., Yeste, M., and Barranco, I. (2020). Glutathione S-Transferases Play a Crucial Role in Mitochondrial Function, Plasma Membrane Stability and Oxidative Regulation of Mammalian Sperm. Antioxidants, 9.","DOI":"10.3390\/antiox9020100"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1038\/onc.2013.548","article-title":"LGALS3BP, lectin galactoside-binding soluble 3 binding protein, promotes oncogenic cellular events impeded by antibody intervention","volume":"34","author":"Stampolidis","year":"2015","journal-title":"Oncogene"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1002\/pros.21287","article-title":"Co-purification of Mac-2 binding protein with galectin-3 and association with prostasomes in human semen","volume":"71","author":"Block","year":"2011","journal-title":"Prostate"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1111\/j.2047-2927.2012.00059.x","article-title":"Comparative proteomic analysis coupled with conventional protein assay as a strategy to identify predictors of successful testicular sperm extraction in patients with non-obstructive azoospermia","volume":"1","author":"Freour","year":"2013","journal-title":"Andrology"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s12610-023-00184-0","article-title":"Seminal plasma biomarkers for predicting successful sperm retrieval in patients with nonobstructive azoospermia: A narrative review of human studies","volume":"33","author":"Li","year":"2023","journal-title":"Basic Clin. Androl."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"10346","DOI":"10.1074\/jbc.M310964200","article-title":"Protease Nexin-1 Inhibits Plasminogen Activation-induced Apoptosis of Adherent Cells","volume":"279","author":"Rossignol","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3029","DOI":"10.1073\/pnas.051630698","article-title":"Male fertility defects in mice lacking the serine protease inhibitor protease nexin-1","volume":"98","author":"Murer","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1095\/biolreprod.110.085100","article-title":"SERPINE2, a serine protease inhibitor extensively expressed in adult male mouse reproductive tissues, may serve as a murine sperm decapacitation factor","volume":"84","author":"Lu","year":"2011","journal-title":"Biol. Reprod."},{"key":"ref_55","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_56","doi-asserted-by":"crossref","unstructured":"Durfey, C.L., Swistek, S.E., Liao, S.F., Crenshaw, M.A., Clemente, H.J., Thirumalai, R.V.K.G., Steadman, C.S., Ryan, P.L., Willard, S.T., and Feugang, J.M. (2019). Nanotechnology-based approach for safer enrichment of semen with best spermatozoa. J. Anim. Sci. Biotechnol., 10.","DOI":"10.1186\/s40104-018-0307-4"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Feugang, J.M., Youngblood, R.C., Greene, J.M., Fahad, A.S., Monroe, W.A., Willard, S.T., and Ryan, P.L. (2012). Application of quantum dot nanoparticles for potential non-invasive bio-imaging of mammalian spermatozoa. J. Nanobiotechnology, 10.","DOI":"10.1186\/1477-3155-10-45"}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/7\/456\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:10:04Z","timestamp":1760033404000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/7\/456"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,15]]},"references-count":57,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["bios15070456"],"URL":"https:\/\/doi.org\/10.3390\/bios15070456","relation":{},"ISSN":["2079-6374"],"issn-type":[{"type":"electronic","value":"2079-6374"}],"subject":[],"published":{"date-parts":[[2025,7,15]]}}}