{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T15:28:04Z","timestamp":1769700484251,"version":"3.49.0"},"reference-count":49,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T00:00:00Z","timestamp":1769472000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012190","name":"Ministry of Science and Higher Education of the Russian Federation","doi-asserted-by":"crossref","award":["FMRM-2025-0032"],"award-info":[{"award-number":["FMRM-2025-0032"]}],"id":[{"id":"10.13039\/501100012190","id-type":"DOI","asserted-by":"crossref"}]},{"name":"RUDN University Scientific Projects Grant System","award":["202789-2-000"],"award-info":[{"award-number":["202789-2-000"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>In this study, the biocatalytic activity of four steroid-transforming strains isolated from the African frog Xenopus laevis and identified as Streptomyces rochei towards pregnane steroids has been investigated. All the isolated strains facilitated the reduction of the C20-carbonyl group and the structures of the metabolites were confirmed by mass spectrometric (MS) and 1H NMR spectroscopic analyses. Hydrocortisone and progesterone were poorly transformed by the streptomycete strains, whereas cortexolone (Reichstein\u2019s substance S) was effectively biotransformed, yielding more than 90% of 17\u03b1,20\u03b2,21\u03b1-trihydroxy-4-pregnen-3-one (20\u03b2-S). Primarily, 20\u03b1-reduction was detected when the microbial isolates were incubated with 17\u03b1-hydroxyprogesterone with the yield of 17\u03b1,20\u03b1-dihydroxy-4-pregnen-3-one (17,20\u03b1-P) reaching 70%. The biological activity of 20\u03b2-S was evaluated in Danio rerio. The results demonstrated that 20\u03b2-S modulated stress- and anxiety-related behavioral responses and activated Pgr-dependent transcriptional pathways in the brain and ovarian tissues. These observations support the potential relevance of the synthesized progestin as a functional regulator in teleost physiology. The findings enhance our understanding of the biodiversity of steroid-transforming actinomycetes inhabiting amphibians and can be successfully employed for the effective microbiological synthesis of biologically active 20-hydroxylated progestins that serve as bioregulators in teleosts.<\/jats:p>","DOI":"10.3390\/biom16020196","type":"journal-article","created":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T15:41:56Z","timestamp":1769528516000},"page":"196","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Microbial Synthesis and Biological Activity of 20\u03b2-Hydroxylated Progestins: Ovarian and Neural Action of 17\u03b1,20\u03b2,21\u03b1-Trihydroxy-4-Pregnen-3-One in Danio rerio"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5623-2930","authenticated-orcid":false,"given":"Vyacheslav V.","family":"Kollerov","sequence":"first","affiliation":[{"name":"Federal Research Center \u00abPushchino Center for Biological Research\u00bb of Russian Academy of Sciences, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Prospekt Nauki, 5, 142290 Pushchino, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vsevolod V.","family":"Pavshintsev","sequence":"additional","affiliation":[{"name":"Institute of Environmental Engineering, RUDN University, 6 Miklukho-Maklaya Street, 117198 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexey V.","family":"Kazantsev","sequence":"additional","affiliation":[{"name":"Chemical Department, Moscow State University, GSP-1, Leninskiye Gori, 1, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9501-8072","authenticated-orcid":false,"given":"Andrei A.","family":"Shutov","sequence":"additional","affiliation":[{"name":"Federal Research Center \u00abPushchino Center for Biological Research\u00bb of Russian Academy of Sciences, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Prospekt Nauki, 5, 142290 Pushchino, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aleksey A.","family":"Vatlin","sequence":"additional","affiliation":[{"name":"Institute of Environmental Engineering, RUDN University, 6 Miklukho-Maklaya Street, 117198 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1014-9250","authenticated-orcid":false,"given":"Nikita A.","family":"Mitkin","sequence":"additional","affiliation":[{"name":"Collagel LLC, Belovezskaya 55, 121353 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olga V.","family":"Fadeeva","sequence":"additional","affiliation":[{"name":"Institute of Mitoengineering MSU LLC, Leninskie Gory, 1, Bld. 73, 119234 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7949-7974","authenticated-orcid":false,"given":"Maxim L.","family":"Lovat","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, Faculty of Biology, Moscow State University, Leninskiye Gori, 1, 119234 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9699-1438","authenticated-orcid":false,"given":"Elena O.","family":"Morgun","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine, Pirogov Russian National Research Medical University, Ostrovityanova Street, 1, 117997 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9917-8428","authenticated-orcid":false,"given":"Marina V.","family":"Donova","sequence":"additional","affiliation":[{"name":"Federal Research Center \u00abPushchino Center for Biological Research\u00bb of Russian Academy of Sciences, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Prospekt Nauki, 5, 142290 Pushchino, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rui, S., Fengrui, G., Yining, Z., Hong, S., Xuewen, Y., Changping, W., and Chunjia, Y. (2025). Biological activity of secondary metabolites of actinomycetes and their potential sources as antineoplastic drugs: A review. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2025.1550516"},{"key":"ref_2","first-page":"1730","article-title":"A review on actinomycetes and their biotechnological application","volume":"4","author":"Dilip","year":"2013","journal-title":"Int. J. Pharm. Sci. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1007\/s10482-024-01964-y","article-title":"Biotechnological potential of actinomycetes in the 21st century: A brief review","volume":"117","author":"Feitoza","year":"2024","journal-title":"Antonie Leeuwenhoek"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"316","DOI":"10.4172\/jpb.1000456","article-title":"Actinomycetes: A source of industrially important enzymes","volume":"10","author":"Mukhtar","year":"2017","journal-title":"J. Proteom. Bioinform."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"210","DOI":"10.3390\/md7020210","article-title":"Antitumor compounds from marine actinomycetes","volume":"7","author":"Olano","year":"2009","journal-title":"Mar. Drugs"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1134\/S0003683821090064","article-title":"Streptomyces tsukubensis VKM Ac-2618D-an effective producer of tacrolimus","volume":"57","author":"Poshekhontseva","year":"2021","journal-title":"Appl. Biochem. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1134\/S0003683807010012","article-title":"Transformation of steroids by actinobacteria: A review","volume":"43","author":"Donova","year":"2007","journal-title":"Appl. Biochem. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1007\/s00253-012-4078-0","article-title":"Microbial steroid transformations: Current state and prospects","volume":"6","author":"Donova","year":"2012","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.molcatb.2008.09.003","article-title":"Transformation of C19-steroids and testosterone production by sterol-transforming strains of Mycobacterium spp.","volume":"57","author":"Egorova","year":"2009","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1186\/s40643-022-00602-7","article-title":"Bioproduction of testosterone from phytosterol by Mycolicibacterium neoaurum strains: \u201cone-pot\u201d, two modes","volume":"9","author":"Tekucheva","year":"2022","journal-title":"Bioresour. Bioprocess."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/j.steroids.2010.06.008","article-title":"Biotransformation of dehydroepiandrosterone (DHEA) with Penicillium griseopurpureum Smith and Penicillium glabrum (Wehmer) Westling","volume":"75","author":"Huang","year":"2010","journal-title":"Steroids"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8153","DOI":"10.1007\/s00253-018-9239-3","article-title":"Bacterial steroid hydroxylases: Enzyme classes, their functions and comparison of their catalytic mechanisms","volume":"102","author":"Szaleniec","year":"2018","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"9496","DOI":"10.1021\/acs.jafc.0c03360","article-title":"Identification, biological characteristics, and active site residues of 3-ketosteroid \u03b41-dehydrogenase homologues from arthrobacter simplex","volume":"68","author":"Luo","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Costa, S., Zappaterra, F., Summa, D., Semeraro, B., and Fantin, G. (2020). \u03941-dehydrogenation and C20 reduction of cortisone and hydrocortisone catalyzed by Rhodococcus strains. Molecules, 25.","DOI":"10.3390\/molecules25092192"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Fufaeva, S.R., Dovbnya, D.V., Ivashina, T.V., Shutov, A.A., and Donova, M.V. (2023). Reconstruction of the steroid 1(2)-dehydrogenation system from Nocardioides simplex VKM Ac-2033D in Mycolicibacterium hosts. Microorganisms, 11.","DOI":"10.3390\/microorganisms11112720"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1093\/oxfordjournals.jbchem.a123150","article-title":"Steroid transhydrogenase activity of 3-ketosteroid-\u03b4-dehydrogenase from Nocardia corallina","volume":"108","author":"Itagaki","year":"1990","journal-title":"J. Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.molcatb.2006.04.007","article-title":"Biotransformation of various natural sterols to androstenones by Mycobacterium sp. and some steroid-converting microbial strains","volume":"41","author":"Sripalakit","year":"2006","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Maltseva, P.Y., Plotnitskaya, N.A., and Ivshina, I.B. (2024). Transformation of terpenoids and steroids using actinomycetes of the genus Rhodococcus. Molecules, 29.","DOI":"10.3390\/molecules29143378"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.jsbmb.2004.01.009","article-title":"Extracellular 3\u03b2-hydroxysteroid oxidase of Mycobacterium vaccae VKM Ac-1815D","volume":"91","author":"Nikolayeva","year":"2004","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_20","first-page":"280","article-title":"Production of testosterone from phytosterol using a single-step microbial transformation by a mutant of Mycobacterium sp.","volume":"28","author":"Lo","year":"2002","journal-title":"J. Ind. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1111\/1751-7915.12433","article-title":"Engineering Mycobacterium smegmatis for testosterone production","volume":"10","author":"Galan","year":"2017","journal-title":"Microb. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1007\/s00253-011-3382-4","article-title":"Transformation of prednisolone to a 20\u03b2-hydroxy prednisolone compound by Streptomyces roseochromogenes TS79","volume":"92","author":"Zhang","year":"2011","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"737315","DOI":"10.1016\/j.aquaculture.2021.737315","article-title":"Maturation inducing hormones in teleosts: Are progestogens always the first to be nominated?","volume":"546","author":"Bulteau","year":"2022","journal-title":"Aquac. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"113842","DOI":"10.1016\/j.ygcen.2021.113842","article-title":"Nuclear progesterone receptor regulates ptger4b and PLA2G4A expression in zebrafish (Danio rerio) ovulation","volume":"311","author":"Baker","year":"2021","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1089\/zeb.2025.0020","article-title":"A novel method to make zebrafish mate by adding a membrane progestin receptor agonist to the water without injection","volume":"22","author":"Ahamed","year":"2025","journal-title":"Zebrafish"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"J\u00e9hannet, P., Palstra, A.P., Meijerhof, M., Schipper, H., Gim\u00e9nez, I.N., Dirks, R.P., Swinkels, W., Heinsbroek, L.T.N., and Komen, H. (2023). Induction of oocyte maturation and ovulation in the European eel (Anguilla anguilla): Comparison of progesterone and 17\u03b1,20\u03b2-dihydroxy-4-pregnen-3-one. Front. Physiol., 14.","DOI":"10.3389\/fphys.2023.1207542"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ygcen.2013.01.005","article-title":"A progestin (17\u03b1,20\u03b2-dihydroxy-4-pregnen-3-one) stimulates early stages of spermatogenesis in zebrafish","volume":"185","author":"Chen","year":"2013","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s10695-025-01519-6","article-title":"Effects of progestogen neurosteroids on locomotor activity in zebrafish embryos and larvae","volume":"51","author":"Mathouchanh","year":"2025","journal-title":"Fish. Physiol. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Silva, A.C., Zubizarreta, L., and Quintana, L. (2020). A teleost fish model to understand hormonal mechanisms of non-breeding territorial behaviour. Front. Endocrinol., 11.","DOI":"10.3389\/fendo.2020.00468"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Zubizarreta, L., Jalabert, C., Silva, A.C., Soma, K.K., and Quintana, L. (2023). Brain and circulating steroids in an electric fish: Relevance for non-breeding aggression. PLoS ONE, 18.","DOI":"10.1101\/2023.07.20.549867"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.ymben.2016.02.010","article-title":"Genetic engineering of Bacillus megaterium for high-yield production of the major teleost progestogens 17\u03b1,20\u03b2-di- and 17\u03b1,20\u03b2,21\u03b1-trihydroxy-4-pregnen-3-one","volume":"36","author":"Gerber","year":"2016","journal-title":"Metab. Eng."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1194\/jlr.M074914","article-title":"Identification and characterization of a 20\u03b2-HSDH from the anaerobic gut bacterium Butyricicoccus desmolans ATCC 43058","volume":"58","author":"Devendran","year":"2017","journal-title":"J. Lipid Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12040","DOI":"10.1074\/jbc.RA119.009390","article-title":"Structural and biochemical characterization of 20\u03b2-hydroxysteroid dehydrogenase from Bifidobacterium adolescentis strain L2-32","volume":"294","author":"Doden","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e00709-25","DOI":"10.1128\/mra.00709-25","article-title":"Draft genome sequence of a steroid-transforming actinomycete isolated from African clawed frog Xenopus laevis","volume":"14","author":"Kollerov","year":"2025","journal-title":"Microbiol. Resour. Announc."},{"key":"ref_35","first-page":"1","article-title":"Isolation of microbes from the skin of terrestrial frogs","volume":"14","author":"Caty","year":"2024","journal-title":"Protocols.io"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.funbio.2021.11.002","article-title":"Steroid modification by filamentous fungus Drechslera sp.: Focus on 7-hydroxylase and 17\u03b2-hydroxysteroid dehydrogenase activities","volume":"126","author":"Kollerov","year":"2022","journal-title":"Fungal Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1016\/j.pnpbp.2011.01.006","article-title":"Pharmacological analysis of zebrafish (Danio rerio) scototaxis","volume":"35","author":"Maximino","year":"2011","journal-title":"Prog. Neuro-Psychopharmacol. Biol. Psychiatry"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"43","DOI":"10.46867\/IJCP.2010.23.01.06","article-title":"Zebrafish behavior in novel environments: Effects of anxiolytic compounds and strain choice","volume":"23","author":"Sackerman","year":"2010","journal-title":"Int. J. Comp. Psychol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e7546","DOI":"10.7717\/peerj.7546","article-title":"Anxiety-like behavior and whole-body cortisol responses to components of energy drinks in zebrafish (Danio rerio)","volume":"7","author":"Alia","year":"2019","journal-title":"PeerJ"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"10064","DOI":"10.1073\/pnas.88.22.10064","article-title":"Three-dimensional structure of holo 3alpha,20beta-hydroxysteroid dehydrogenase: A member of a short-chain dehydrogenase family","volume":"88","author":"Ghosh","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"105874","DOI":"10.1016\/j.jsbmb.2021.105874","article-title":"Functional characterization of two 20\u03b2-hydroxysteroid dehydrogenase type 2 homeologs from Xenopus laevis reveals multispecificity","volume":"210","author":"Tokarz","year":"2021","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/S0039-128X(03)00043-6","article-title":"Microbial conversion of pregna-4,9(11)-diene-17\u03b1,21-diol-3,20-dione acetates by Nocardioides simplex VKM Ac-2033D","volume":"68","author":"Fokina","year":"2003","journal-title":"Steroids"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/S0960-0760(99)00132-6","article-title":"Microbial transformations of steroids-XI. Progesterone transformation by Streptomyces roseochromogenes\u2014Purification and characterisation of the 16alpha-hydroxylase system","volume":"71","author":"Berrie","year":"1999","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/jobm.3620350102","article-title":"Transformation reactions of progesterone by different species of Streptomyces","volume":"35","author":"Atta","year":"1995","journal-title":"J. Basic Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1016\/S0022-2275(20)37722-1","article-title":"Mode of action of steroid desmolase and reductases synthesized by Clostridium \u201cscindens\u201d (formerly Clostridium strain 19)","volume":"25","author":"Winter","year":"1984","journal-title":"J. Lipid Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"111174","DOI":"10.1016\/j.mce.2021.111174","article-title":"Gut feelings about bacterial steroid-17,20-desmolase","volume":"525","author":"Ly","year":"2021","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1073\/pnas.0137017100","article-title":"Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis","volume":"100","author":"Yang","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Diotel, N., Charlier, T.D., Lefebvre d\u2019Hellencourt, C., Couret, D., Trudeau, V.L., Nicolau, J.C., Meilhac, O., Kah, O., and Pellegrini, E. (2018). Steroid transport, local synthesis, and signaling within the brain: Roles in neurogenesis, neuroprotection, and sexual behaviors. Front. Neurosci., 12.","DOI":"10.3389\/fnins.2018.00084"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Vaudry, H., Do Rego, J.L., Burel, D., Luu-Th\u00e9, V., Pelletier, G., and Tsutsui, K. (2011). Neurosteroid biosynthesis in the brain of amphibians. Front. Endocrinol., 2.","DOI":"10.3389\/fendo.2011.00079"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/2\/196\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T05:25:14Z","timestamp":1769664314000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/2\/196"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,27]]},"references-count":49,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["biom16020196"],"URL":"https:\/\/doi.org\/10.3390\/biom16020196","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,27]]}}}