{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T19:05:50Z","timestamp":1770750350511,"version":"3.50.0"},"reference-count":38,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T00:00:00Z","timestamp":1770681600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002923","name":"CONICET Argentina","doi-asserted-by":"publisher","award":["PIP 0012-2023"],"award-info":[{"award-number":["PIP 0012-2023"]}],"id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003074","name":"Universidad Nacional de Cuyo","doi-asserted-by":"publisher","award":["FONCYT\/PICT 2021-GRF-TII00279"],"award-info":[{"award-number":["FONCYT\/PICT 2021-GRF-TII00279"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003074","name":"Universidad Nacional de Cuyo","doi-asserted-by":"publisher","award":["FONCYT\/PICT 02642-2018"],"award-info":[{"award-number":["FONCYT\/PICT 02642-2018"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003074","name":"Universidad Nacional de Cuyo","doi-asserted-by":"publisher","award":["FONCYT\/PICT 01762-2019"],"award-info":[{"award-number":["FONCYT\/PICT 01762-2019"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003074","name":"Universidad Nacional de Cuyo","doi-asserted-by":"publisher","award":["SIIP 2025 80020240100092UN"],"award-info":[{"award-number":["SIIP 2025 80020240100092UN"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Universidad del Aconcagua","award":["CIUDA 2025"],"award-info":[{"award-number":["CIUDA 2025"]}]},{"name":"Universidad del Aconcagua","award":["DICYT 2025 and 2024"],"award-info":[{"award-number":["DICYT 2025 and 2024"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hypoprolactinemia (hypoPRL) disrupts lactation and compromises milk production. Although maternal milk is a critical source of nutrients and bioactive compounds for newborns, the consequences of deficient lactation based on reduced milk quantity on the offspring\u2019s immune development remain incompletely understood. Therefore, this study aimed to elucidate how deficient lactation due to hypoPRL interferes with offspring immunity and development. Female Sprague Dawley (SD) and spontaneous hypoPRL Oncins France Colony A (OFA) rats were euthanized on day 2 of lactation to assess the impact of hypoPRL on serum, milk, and tissue samples. We demonstrated that early deficient lactation in the OFA model impaired maternal performance, leading to increased pup mortality during early lactation. OFA pups exhibited reduced body weight and weight gain, decreased cerebral weight and index, and an increased cephalization index. Thymic development was markedly altered, as evidenced by reduced thymus weight, area, and cortical extension. These structural changes were accompanied by increased thymic Rag 1 expression, suggesting altered lymphocyte development. In parallel, passive immune transfer was compromised, with reduced levels of OVA-specific immunoglobulin isotypes detected in pup serum, reflecting changes in milk-derived immune support. In summary, maternal hypoPRL during early lactation adversely affects offspring growth and thymic maturation with lasting consequences for neonatal immune maturation. This study provides a novel experimental framework to investigate the consequences of lactation deficiency with potential implications for understanding lactation insufficiency and its impact on neonatal immune exposure.<\/jats:p>","DOI":"10.3390\/ijms27041708","type":"journal-article","created":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T11:16:38Z","timestamp":1770722198000},"page":"1708","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Early Deficient Lactation Differentially Affects Neonatal Thymic Cortical Development and Humoral Immune Responses in Rats"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2502-8248","authenticated-orcid":false,"given":"Mar\u00eda Bel\u00e9n","family":"S\u00e1nchez","sequence":"first","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Facultad de Ciencias Veterinarias y Ambientales, Universidad Juan Agust\u00edn Maza, Guaymall\u00e9n, Mendoza CP 5519, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7616-6638","authenticated-orcid":false,"given":"Mar\u00eda Cecilia","family":"Michel Lara","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Facultad de Farmacia y Bioqu\u00edmica, Universidad Juan Agust\u00edn Maza, Guaymall\u00e9n, Mendoza CP 5519, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4845-1738","authenticated-orcid":false,"given":"Mar\u00eda Jos\u00e9","family":"German\u00f3","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias M\u00e9dicas, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Flavia Judith","family":"Neira","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Facultad de Kinesiolog\u00eda, Universidad Juan Agust\u00edn Maza, Guaymall\u00e9n, Mendoza CP 5519, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciana Bel\u00e9n","family":"Viruel","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Facultad de Ciencias Veterinarias y Ambientales, Universidad Juan Agust\u00edn Maza, Guaymall\u00e9n, Mendoza CP 5519, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3661-9741","authenticated-orcid":false,"given":"Jacqueline Lisset","family":"Tomsich","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1348-8012","authenticated-orcid":false,"given":"Claudio","family":"Rodr\u00edguez-Camejo","sequence":"additional","affiliation":[{"name":"Laboratorio de Inmunolog\u00eda, Facultad de Ciencias\/Facultad de Qu\u00edmica, Universidad de la Rep\u00fablica, Instituto de Higiene, Montevideo CP 11200, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariana","family":"Troncoso","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias Exactas y Naturales, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa Olivia","family":"Pietrobon","sequence":"additional","affiliation":[{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias M\u00e9dicas, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2246-1393","authenticated-orcid":false,"given":"Marta","family":"Soaje","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias M\u00e9dicas, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Laboratorio de Inmunolog\u00eda, Facultad de Ciencias\/Facultad de Qu\u00edmica, Universidad de la Rep\u00fablica, Instituto de Higiene, Montevideo CP 11200, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4310-6684","authenticated-orcid":false,"given":"Evelyn L.","family":"Jara","sequence":"additional","affiliation":[{"name":"Departamento de Farmacolog\u00eda, Facultad de Ciencias Biol\u00f3gicas, Universidad de Concepci\u00f3n, Concepci\u00f3n CP 4070386, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susana Ruth","family":"Valdez","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias Exactas y Naturales, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9437-0579","authenticated-orcid":false,"given":"Juan Pablo","family":"Mackern-Oberti","sequence":"additional","affiliation":[{"name":"Instituto de Medicina y Biolog\u00eda Experimental de Cuyo CONICET, Universidad Nacional de Cuyo, Ciudad, Mendoza CP 5500, Argentina"},{"name":"Universidad Nacional de Cuyo, Facultad de Ciencias M\u00e9dicas, Ciudad, Mendoza CP 5500, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,10]]},"reference":[{"key":"ref_1","unstructured":"WHO (2018). Guideline: Counselling of Women to Improve Breastfeeding Practices, WHO."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1016\/S0140-6736(15)01024-7","article-title":"Breastfeeding in the 21st century: Epidemiology, mechanisms, and lifelong effect","volume":"387","author":"Victora","year":"2016","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.pcl.2012.10.002","article-title":"Human milk composition: Nutrients and bioactive factors","volume":"60","author":"Ballard","year":"2013","journal-title":"Pediatr. Clin. N. Am."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Azagra-Boronat, I., Tres, A., Massot-Cladera, M., Franch, A., Castell, M., Guardiola, F., Perez-Cano, F.J., and Rodriguez-Lagunas, M.J. (2020). Associations of Breast Milk Microbiota, Immune Factors, and Fatty Acids in the Rat Mother-Offspring Pair. Nutrients, 12.","DOI":"10.3390\/nu12020319"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2247","DOI":"10.1111\/all.16054","article-title":"Colostrum is required for the postnatal ontogeny of small intestine innate lymphoid type 2 cells and successful anti-helminth defences","volume":"79","author":"Rekima","year":"2024","journal-title":"Allergy"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Garofoli, F., Civardi, E., Pisoni, C., Angelini, M., and Ghirardello, S. (2023). Anti-Inflammatory and Anti-Allergic Properties of Colostrum from Mothers of Full-Term and Preterm Babies: The Importance of Maternal Lactation in the First Days. Nutrients, 15.","DOI":"10.3390\/nu15194249"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Abdulla, F., Hossain, M.M., Karimuzzaman, M., Ali, M., and Rahman, A. (2022). Likelihood of infectious diseases due to lack of exclusive breastfeeding among infants in Bangladesh. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0263890"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1093\/ajcn\/80.3.722","article-title":"Improved thymic function in exclusively breastfed infants is associated with higher interleukin 7 concentrations in their mothers\u2019 breast milk","volume":"80","author":"Ngom","year":"2004","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kashiwagi, H., Seki, T., Oshima, S., Ohno, Y., Shimizu, T., Yamada, S., Katano, N., Goto, Y., Yasuda, A., and Tsuda, B. (2022). High-progesterone environment preserves T cell competency by evading glucocorticoid effects on immune regulation. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.1000728"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1016\/j.autrev.2018.03.006","article-title":"Contribution of sex steroids and prolactin to the modulation of T and B cells during autoimmunity","volume":"17","author":"Recalde","year":"2018","journal-title":"Autoimmun. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1038\/s41574-022-00742-y","article-title":"Hormonal regulation of mammary gland development and lactation","volume":"19","author":"Hannan","year":"2023","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"111084","DOI":"10.1016\/j.mce.2020.111084","article-title":"Three lactation-related hormones: Regulation of hypothalamus-pituitary axis and function on lactation","volume":"520","author":"Ni","year":"2021","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/s10911-016-9369-0","article-title":"Role of Prolactin in Promotion of Immune Cell Migration into the Mammary Gland","volume":"22","author":"Dill","year":"2017","journal-title":"J. Mammary Gland Biol. Neoplasia"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4919","DOI":"10.1113\/jphysiol.2009.176289","article-title":"Maternal prolactin inhibition during lactation programs for metabolic syndrome in adult progeny","volume":"587","author":"Bonomo","year":"2009","journal-title":"J. Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e3021","DOI":"10.1210\/clinem\/dgab201","article-title":"PRL Mutation Causing Alactogenesis: Insights Into Prolactin Structure and Function Relationships","volume":"106","author":"Moriwaki","year":"2021","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"361","DOI":"10.3109\/10253890.2011.624223","article-title":"Lactation deficit in OFA hr\/hr rats may be caused by differential sensitivity to stress compared with Wistar and Sprague Dawley rats","volume":"15","author":"Valdez","year":"2012","journal-title":"Stress"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1530\/REP-06-0032","article-title":"Hormonal profile and reproductive performance in lactation deficient (OFA hr\/hr) and normal (Sprague-Dawley) female rats","volume":"133","author":"Valdez","year":"2007","journal-title":"Reproduction"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1530\/REP-12-0387","article-title":"Impaired mammary gland T cell population during early lactation in hypoprolactinemic lactation-deficient rats","volume":"146","author":"Valdez","year":"2013","journal-title":"Reproduction"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Georgescu, T., Swart, J.M., Grattan, D.R., and Brown, R.S.E. (2021). The Prolactin Family of Hormones as Regulators of Maternal Mood and Behavior. Front. Glob. Womens Health, 2.","DOI":"10.3389\/fgwh.2021.767467"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.jneuroim.2004.01.012","article-title":"Postpartum maternal corticosterone decreases maternal and neonatal antibody levels and increases the susceptibility of newborn mice to herpes simplex virus-associated mortality","volume":"150","author":"Yorty","year":"2004","journal-title":"J. Neuroimmunol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Pournasiri, Z., Bakhtiary, M., Nikparast, A., Hashemi, S.M., Narjes Ahmadizadeh, S., Behzad, A., and Asghari, G. (2024). The association between nutritional status measured by body mass index and outcomes in the pediatric intensive care unit. Front. Pediatr., 12.","DOI":"10.3389\/fped.2024.1421155"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1002\/uog.9008","article-title":"Neurodevelopment following fetal growth restriction and its relationship with antepartum parameters of placental dysfunction","volume":"37","author":"Baschat","year":"2011","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S2213-8587(15)00327-7","article-title":"Association of maternal thyroid function during early pregnancy with offspring IQ and brain morphology in childhood: A population-based prospective cohort study","volume":"4","author":"Korevaar","year":"2016","journal-title":"Lancet Diabetes Endocrinol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"112370","DOI":"10.1016\/j.mce.2024.112370","article-title":"Hyperthyroidism keeps immunoglobulin levels but reduces milk fat and CD11b\/c(+) cells on early lactation","volume":"594","author":"Sanchez","year":"2024","journal-title":"Mol. Cell Endocrinol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1001\/jamapediatrics.2017.1579","article-title":"Association of Neonatal Glycemia With Neurodevelopmental Outcomes at 4.5 Years","volume":"171","author":"McKinlay","year":"2017","journal-title":"JAMA Pediatr."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Nabukeera-Barungi, N., Lanyero, B., Grenov, B., Friis, H., Namusoke, H., Mupere, E., Michaelsen, K.F., Molgaard, C., Wiese, M., and Nielsen, D.S. (2021). Thymus size and its correlates among children admitted with severe acute malnutrition: A cross-sectional study in Uganda. BMC Pediatr., 21.","DOI":"10.1186\/s12887-020-02457-3"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"231","DOI":"10.37897\/RJP.2024.4.3","article-title":"Comparison of thymus size in normal versus malnourished children","volume":"73","author":"Muthu","year":"2024","journal-title":"Rom. J. Pediatr."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Savino, W., Duraes, J., Maldonado-Galdeano, C., Perdigon, G., Mendes-da-Cruz, D.A., and Cuervo, P. (2022). Thymus, undernutrition, and infection: Approaching cellular and molecular interactions. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.948488"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1126\/science.1831564","article-title":"Thymocyte expression of RAG-1 and RAG-2: Termination by T cell receptor cross-linking","volume":"253","author":"Turka","year":"1991","journal-title":"Science"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"442","DOI":"10.3390\/nu3040442","article-title":"Perspectives on immunoglobulins in colostrum and milk","volume":"3","author":"Hurley","year":"2011","journal-title":"Nutrients"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Grases-Pinto, B., Abril-Gil, M., Torres-Castro, P., Castell, M., Rodriguez-Lagunas, M.J., Perez-Cano, F.J., and Franch, A. (2021). Rat Milk and Plasma Immunological Profile throughout Lactation. Nutrients, 13.","DOI":"10.3390\/nu13041257"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Dias-Rocha, C.P., Costa, J.C.B., Oliveira, Y.S., Fassarella, L.B., Woyames, J., Atella, G.C., Santos, G.R.C., Pereira, H.M.G., Pazos-Moura, C.C., and Almeida, M.M. (2023). Maternal high-fat diet decreases milk endocannabinoids with sex-specific changes in the cannabinoid and dopamine signaling and food preference in rat offspring. Front. Endocrinol., 14.","DOI":"10.3389\/fendo.2023.1087999"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e995","DOI":"10.1002\/cpz1.995","article-title":"Mouse and Rat Anesthesia and Analgesia","volume":"4","author":"Oh","year":"2024","journal-title":"Curr. Protoc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e00631","DOI":"10.1002\/brb3.631","article-title":"Learning and memory disabilities in IUGR babies: Functional and molecular analysis in a rat model","volume":"7","author":"Krauel","year":"2017","journal-title":"Brain Behav."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1007\/BF02990063","article-title":"Effect of chronic thyroid hormone treatment on cycling, ovulation, serum reproductive hormones and ovarian LH and prolactin receptors in rats","volume":"3","author":"Jahn","year":"1995","journal-title":"Endocrine"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Gangisetty, O., Wynne, O., Jabbar, S., Nasello, C., and Sarkar, D.K. (2015). Fetal Alcohol Exposure Reduces Dopamine Receptor D2 and Increases Pituitary Weight and Prolactin Production via Epigenetic Mechanisms. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0140699"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Zschemisch, N.H., Glage, S., Wedekind, D., Weinstein, E.J., Cui, X., Dorsch, M., and Hedrich, H.J. (2012). Zinc-finger nuclease mediated disruption of Rag1 in the LEW\/Ztm rat. BMC Immunol., 13.","DOI":"10.1186\/1471-2172-13-60"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/4\/1708\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T11:22:41Z","timestamp":1770722561000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/4\/1708"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,10]]},"references-count":38,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["ijms27041708"],"URL":"https:\/\/doi.org\/10.3390\/ijms27041708","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,10]]}}}