Journal of Steroid Biochemistry and Molecular Biology最新文献

{"title":"Determination of steroid reference intervals in a pregnancy population","authors":"Marius Lahti-Pulkkinen ,&nbsp;Katri Räikkönen ,&nbsp;Agnieszka Basiukajc ,&nbsp;Patricia Lee ,&nbsp;Scott G. Denham ,&nbsp;Joanna P. Simpson ,&nbsp;Pia Villa ,&nbsp;Esa Hämäläinen ,&nbsp;Hannele Laivuori ,&nbsp;Eero Kajantie ,&nbsp;Kati Heinonen ,&nbsp;Polina Girchenko ,&nbsp;Rebecca M. Reynolds ,&nbsp;Natalie ZM Homer","doi":"10.1016/j.jsbmb.2025.106691","DOIUrl":"10.1016/j.jsbmb.2025.106691","url":null,"abstract":"<div><div>Steroids, including mineralocorticoids, glucocorticoids, and sex hormones play a critical role in pregnancy. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis offers an opportunity to conduct simultaneous multiplex steroid analysis within a given sample. This paper describes the LC-MS/MS steroid analysis method developed for assessing plasma specific reference ranges of 18 steroids from plasma samples (200 µL) of pregnant women participating in the PREDO study. Samples were prepared using supported liquid extraction and analyzed on an Acquity I-Class UPLC and a QTrap6500 + mass spectrometer. Mass spectrometry parameters were optimized for each steroid and chromatographic separation of 18 steroids was developed. Changes in steroid levels across pregnancy were assessed. Samples were collected after an overnight fast between 07:00 and 09:00. Data from 257 samples from 96 women with uncomplicated pregnancy (women with pre-pregnancy normal weight and no diabetes or hypertensive disorders before or during pregnancy, who delivered a live child at ≥ 37 weeks of gestation with appropriate for gestational age birth weight) were analyzed to calculate steroid reference ranges over three time points, between 11.6 and 14.3, 17.7–22.9, and 25.6–29.0 pregnancy weeks. Levels of progestogens, glucocorticoids, mineralocorticoids, estrogens, their precursors, and metabolites increased significantly across pregnancy. Androgen levels remained stable, except for a decrease in 5α-dihydrotestosterone. The LC-MS/MS method also showed validity in analyses of 917 samples of 328 women with complicated pregnancies. The method is suitable for the analysis of 18 steroids in plasma during pregnancy and the investigation of pregnancy complications, fetal growth, and development after birth.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106691"},"PeriodicalIF":2.7,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Supportive effect of corticosteroid on bone marrow recovery in FLT3/ITD positive acute myeloid leukemia with trisomy 13","authors":"Rie Tabata ,&nbsp;Naoki Yamamoto ,&nbsp;Chiharu Tabata","doi":"10.1016/j.jsbmb.2025.106697","DOIUrl":"10.1016/j.jsbmb.2025.106697","url":null,"abstract":"<div><div>Trisomy 13 is a rare chromosomal abnormality mainly observed in patients with FAB M0, and is associated with increased <em>FLT3</em> expression, characterized by aggressive and inferior outcomes. Herein, we describe a case of <em>FLT3/</em>ITD-positive acute myeloid leukemia with trisomy 13, whose neutropenia and thrombocytopenia were improved by corticosteroids with a decrease in blasts in the peripheral blood without chemotherapy. Only short-term chemotherapy could be administered because of the patient’s poor general condition. In contrast, repeated administration of methylprednisolone pulse therapy followed by high-dose prednisolone was required to control the interstitial pneumonitis. Although appropriate chemotherapy, such as oral administration of FLT inhibitors, could not be administered, a dramatic decrease in blasts and an increase in both neutrophils and platelets were observed. The addition of corticosteroids to chemotherapy is associated with a better clinical course in patients with hyperleukocytic and pediatric acute myeloid leukemia. In the present patient, the peripheral WBC count was not high, the steroids allowed for the recovery of normal hematopoiesis with diminishing blasts. Although stem cell transplantation is necessary to improve overall survival for aggressive acute myeloid leukemia, intensive chemotherapy cannot often be administered in frail older patients. Lower dose of steroids might be able to effectively control acute myeloid leukemia without serious adverse effects, resulting in a better clinical course in frail older patients with acute myeloid leukemia.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106697"},"PeriodicalIF":2.7,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143350481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of 7-dehydrocholesterol and cholesterol in whole blood vs. plasma samples for diagnosis of SLOS","authors":"Zhibin Huang ,&nbsp;Yitao Luo ,&nbsp;Chao Zhang ,&nbsp;Wei Wu ,&nbsp;Yangxi Li ,&nbsp;Li Ma ,&nbsp;Qingxia Zhang ,&nbsp;Yulan Rao ,&nbsp;Chengqiang Zhang","doi":"10.1016/j.jsbmb.2025.106682","DOIUrl":"10.1016/j.jsbmb.2025.106682","url":null,"abstract":"<div><h3>Background</h3><div>Smith-Lemli-Opitz Syndrome (SLOS) is an autosomal recessive disorder characterized by specific clinical features. This study investigated the correlation and transference ratio of 7-dehydrocholesterol (7-DHC) and cholesterol (CHOL) as screening/diagnostic markers for SLOS in whole blood and plasma samples.</div></div><div><h3>Methods</h3><div>The gas chromatography-mass spectrometry (GC-MS) method was employed to analyze whole blood and plasma samples collected from 28 healthy volunteers and one patient with SLOS. Before analysis, an optimized and validated assay for 7-DHC and CHOL in whole blood was developed. Pearson’s correlation analysis was then performed to assess the correlation between the levels of 7-DHC and CHOL in whole blood and plasma.</div></div><div><h3>Results</h3><div>The correlation analysis revealed a significant correlation between the levels of 7-DHC and CHOL in whole blood and plasma, with correlation coefficients (r) greater than 0.5 (0.565 for 7-DHC and 0.692 for CHOL). These findings suggest the feasibility of transference between whole blood and plasma samples.</div></div><div><h3>Conclusions</h3><div>Whole blood can be considered an optimal alternative diagnostic tool for predicting SLOS in carriers, surpassing the reliance on plasma alone. This study introduces a new diagnostic matrix for determining 7-DHC and CHOL, expanding the scope of SLOS diagnosis and research.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106682"},"PeriodicalIF":2.7,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of NR5A2 in regulating sex differentiation, steroidogenesis, and gonadal development in Chlamys farreri","authors":"Lianxue Han ,&nbsp;Jingjing Miao ,&nbsp;Min Ding ,&nbsp;Qichao Fan ,&nbsp;Xuening Wang ,&nbsp;Luqing Pan","doi":"10.1016/j.jsbmb.2025.106690","DOIUrl":"10.1016/j.jsbmb.2025.106690","url":null,"abstract":"<div><div><em>Chlamys farreri</em> is a commercially important bivalve species in global aquaculture. However, research on the mechanisms regulating its sex differentiation and reproduction remains relatively sparse. In this study, the role of nuclear receptor subfamily 5 group A member 2 (<em>NR5A2</em>) in sex differentiation, steroidogenesis, and gonadal development in <em>C. farreri</em> was investigated using a 28-day RNA interference experiment. RNA-seq data analysis revealed differentially expressed genes between males and females following <em>NR5A2</em> knockdown. Weighted gene co-expression network analysis (WGCNA) further identified gene modules closely associated with reproductive development, with the yellow module demonstrating a significant correlation with the sex phenotype. Gene set enrichment analysis (GSEA) identified several signaling pathways related to reproduction that were suppressed, including ovarian follicle development, cholesterol metabolism, and ovarian steroidogenesis. Based on the above analysis, we identified 25 differentially expressed genes linked to these processes. Histological observations revealed that <em>NR5A2</em> knockdown significantly delayed gonadal development in both sexes of scallops, as indicated by a notable decrease in follicular cell number and size. Taken together, <em>NR5A2</em> knockdown significantly affected signaling pathways related to cholesterol metabolism, ovarian steroidogenesis, sex differentiation and gonadal development, providing a novel theoretical basis for understanding sex differentiation and reproductive development in invertebrates.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106690"},"PeriodicalIF":2.7,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143350478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the association between FOK1 polymorphism in the vitamin D receptor (VDR) gene and type 2 diabetes prevalence: A comprehensive analysis","authors":"Romina P. Martinelli ,&nbsp;Candela Petroni ,&nbsp;Josefina Martinez ,&nbsp;Cristina Cuesta ,&nbsp;Luis Esteban ,&nbsp;Alejandra M. Pacchioni ,&nbsp;Pablo Arias","doi":"10.1016/j.jsbmb.2025.106692","DOIUrl":"10.1016/j.jsbmb.2025.106692","url":null,"abstract":"<div><div>There is mounting evidence that suggests vitamin D insufficiency may have a role in the emergence of type 2 diabetes. Additionally, as VDR mediates the actions of vitamin D, variants in its sequence could have implications in this disease. One of these polymorphisms, Fok1 (rs2228570), has been demonstrated to generate changes in the receptor’s structure, causing a shorter protein. The purpose of this research is to establish a potential association between the Fok1 polymorphism and DM2. To achieve such goal, a comprehensive study of this SNP was conducted using functional in-silico analysis and a systematic review with meta-analysis. Additionally, an examination of VDR gene expression in patients with diabetes compared to controls was performed in order to investigate possible differences in expression levels. Our expression analysis showed that VDR has no differential expression between these two groups. To study its functional consequences and stability, different tools were combined, without consistent results. Finally, our systematic review and meta-analysis showed that theFok1 variant was not significantly associated with the DM2 prevalence. This extensive analysis did not provide support for an association between the presence of Fok1 polymorphism and DM2. This result aligns with some previous studies but contrasts others that have reported both protective and risk factors.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106692"},"PeriodicalIF":2.7,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143331772","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"GPER agonist G-1 activates YAP to induce apoptosis in breast cancer cells","authors":"Ze Fu ,&nbsp;Xin Xin ,&nbsp;Yongtong Zhan ,&nbsp;Xuhong Fan ,&nbsp;Xin Li ,&nbsp;Tongsheng Chen ,&nbsp;Xiaoping Wang","doi":"10.1016/j.jsbmb.2025.106693","DOIUrl":"10.1016/j.jsbmb.2025.106693","url":null,"abstract":"<div><div>G-1, a G protein-coupled estrogen receptor (GPER)-specific agonist, exhibits anticancer potential in breast cancer cells. This study aims to explore the molecular basis of apoptosis induced by G-1 in MCF-7 and MDA-MB-231 breast cancer cells. Here, we found that G-1 induced cytotoxicity and GPER-dependent apoptosis with PARP cleavage and mitochondrial membrane potential (MMP) loss, as well as nuclear condensation. Fluorescence resonance energy transfer (FRET) analysis in living cells indicated that G-1 effectively disrupted the interaction between large tumor suppressor 1/2 (LATS1/2) and Yes-associated protein (YAP). Furthermore, G-1 reduced YAP phosphorylation levels and promoted its nuclear accumulation. Notably, knockdown of YAP attenuated G-1-induced apoptosis, highlighting the crucial role of YAP in this process. Additionally, FRET analysis revealed that G-1 enhanced the binding of YAP to p73, leading to an increase in Bcl-2-associated X protein (Bax) expression and an induction of apoptosis. In summary, our findings demonstrate that G-1 induces apoptosis through the GPER/YAP/p73-mediated pathway.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106693"},"PeriodicalIF":2.7,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143366622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Untargeted metabolomics revealed that quercetin improved adrenal gland metabolism disorders and modulated the HPA axis in perimenopausal depression model rats","authors":"Ziran Yu,&nbsp;Chenlu Feng,&nbsp;Ying Chen,&nbsp;Weidi Wang,&nbsp;Xiujuan Zhao","doi":"10.1016/j.jsbmb.2025.106696","DOIUrl":"10.1016/j.jsbmb.2025.106696","url":null,"abstract":"<div><div>Perimenopausal depression is a psychiatric disorder that occurs around the time of menopause and seriously affects women's health. The pathogenesis of perimenopausal depression is unclear which affects its prevention and treatment. Quercetin is a flavonoid compound with antidepressant and estrogen-like effects. The aim of this research was to investigate the role of quercetin on adrenal gland metabolic disorders in perimenopausal depressed rats based on untargeted metabolomics. Female Wistar rats with no difference in sucrose preference were randomly separated into four groups (n = 12): sham-operated group; perimenopausal depression model group; model + 50 mg/kg.bw quercetin group; model + 0.27 mg/kg.bw 17β-estradiol group. After successful modeling, adrenal gland and hypothalamic samples were collected for metabolomics experiments and detection of related indicators. A total of 22 differential metabolites were identified in the model group, and pathway analysis revealed adrenal gland metabolism abnormalities including steroid hormone biosynthesis, arachidonic acid metabolism, and linoleic acid metabolism. Notably, Spearman’s rank correlation analysis between differential metabolites and rat behavioral results showed strong positive or negative correlations (<em>P</em> &lt; 0.01). Meanwhile, the hypothalamus of the model group showed TrkB-BDNF signaling pathway abnormality, and the HPA axis was found to play an important role in perimenopausal depression. Treatment with quercetin or 17β-estradiol restored these abnormal changes. It suggested that quercetin can regulate adrenal metabolic disorders through multiple pathways, thereby ameliorating perimenopausal depression.Further more, quercetin can modulate HPA axis through the TrkB-BDNF signaling pathway. This research provides new ideas for the application of quercetin in the precaution and treatment of perimenopausal depression.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106696"},"PeriodicalIF":2.7,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143331762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Drug repurposing opportunities for breast cancer and seven common subtypes","authors":"Yilong Lin ,&nbsp;Songsong Wang ,&nbsp;Yun Zhang ,&nbsp;Jing She ,&nbsp;Yue Zhang ,&nbsp;Ruidan Zhao ,&nbsp;Zhongquan Qi ,&nbsp;Ruiqin Yang ,&nbsp;Liyi Zhang ,&nbsp;Qingmo Yang","doi":"10.1016/j.jsbmb.2024.106652","DOIUrl":"10.1016/j.jsbmb.2024.106652","url":null,"abstract":"<div><div>Breast cancer is a substantial global health problem, and drug repurposing provides novel opportunities to address the urgent need for therapeutics. According to significant Mendelian randomization (MR) results, we identified 26 genes for overall breast cancer, 25 genes for ER+ breast cancer and 4 genes (CASP8, KCNN4, MYLK4, TNNT3) for ER- breast cancer. In order to explore the differences between 5 intrinsic subtypes, we found 29 actionable druggable genes for Luminal A breast cancer, 2 genes (IGF2 and TNNT3) for Luminal B breast cancer, 1 gene (FAAH) for Luminal B HER2 negative breast cancer, and 3 genes (CASP8, KCNN4, and TP53) for triple-negative breast cancer. After colocalization analysis, we determined OPRL1 as a prioritized target in both overall and Luminal A breast cancer. Additionally, FES and FAAH were considered prioritized targets for ER+ breast cancer. Through molecular docking, crizotinib stand out as a prioritized FES target drug repurposing opportunity with the lowest binding energy (-10.13 kJ·mol⁻¹) and CCK-8 assay showed ER+ cell groups were more sensitive to crizotinib than ER- cell groups. In conclusion, OPRL1 was identified as a prioritized target for both overall and Luminal A breast cancer. Moreover, FES and FAAH were recognized as prioritized targets for ER+ breast cancer.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"246 ","pages":"Article 106652"},"PeriodicalIF":2.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of novel sialylation-associated microRNA signature for prognostic assessment in breast cancer and its implications for the tumor microenvironment","authors":"Yong-Zi Chen ,&nbsp;Shilei Xu ,&nbsp;Hailing Ren ,&nbsp;Jun Zhang ,&nbsp;Yongsheng Jia ,&nbsp;Haiyan Sun","doi":"10.1016/j.jsbmb.2025.106683","DOIUrl":"10.1016/j.jsbmb.2025.106683","url":null,"abstract":"<div><div>Sialylation, a key post-translational modification essential for protein function, is regulated by steroid hormones, along with other glycosylations like fucosylation. These modifications influence tumor growth and metastasis by modulating immune activation. MicroRNAs (miRNAs), crucial in gene expression, affect sialylation and are emerging as promising biomarkers in breast cancer, though their prognostic value remains unclear. Sialylation-related miRNAs were identified through Pearson correlation analysis, and an eight-miRNA risk signature was developed using univariate and Least Absolute Shrinkage and Selection Operator (LASSO) regression in the TCGA dataset. The prognostic value was validated in two independent GEO datasets. Multivariate analysis confirmed that the miRNA risk score is an independent predictor of overall survival (OS). A nomogram integrating clinical characteristics and the risk score was created to predict 1-, 3-, and 5-year OS, assessed through calibration curves, ROC curves, and area under the ROC curve (AUC). Biological pathways were explored using GSEA and GSVA, while immune infiltrates were identified through CIBERSORT and TIMER.</div><div>The eight-miRNA signature effectively predicted OS, recurrence-free survival, and disease-free survival. High-risk patients exhibited increased macrophage and neutrophil levels, indicative of a poor prognosis. High-risk patients, especially those with triple-negative breast cancer, had significantly worse outcomes. This risk score could inform personalized treatment strategies in breast cancer management.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"248 ","pages":"Article 106683"},"PeriodicalIF":2.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143124083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CaCo-2 cell junction derangement exerted by the single addition of oxysterols commonly detected in foods is markedly quenched when they are in mixture","authors":"Noemi Iaia ,&nbsp;Federico Canzoneri ,&nbsp;Fiorella Biasi ,&nbsp;Giuseppe Poli ,&nbsp;Roberto Menta ,&nbsp;Gabriella Testa ,&nbsp;Paola Gamba","doi":"10.1016/j.jsbmb.2024.106648","DOIUrl":"10.1016/j.jsbmb.2024.106648","url":null,"abstract":"<div><div>The selective permeability of the gut epithelial barrier is heavily reliant on the stability of cell junctions, often challenged by a variety of dietary stressors, including non-enzymatic cholesterol oxidation products (COPs). A marked decrease of the tight junctions claudin-1 and occludin, and of the adherens junction E-cadherin was previously detected in differentiated CaCo-2 monolayers challenged by a single addition of 7β-hydroxycholesterol (7βOHC) or 7-ketocholesterol (7KC) in the lowest micromolar range. However, in the diet, oxysterols are occurring in a mixture. Hence, the aim of the present study was to evaluate whether cell incubation with all the main dietary COPs together quench the intercellular junction derangement previously observed as exerted by 7βOHC and 7KC singularly added. Two chocolate prototypes, respectively made with fresh (oxy-Mix1) or six-months stored whole milk powder (oxy-Mix2), were compared. The second prototype showed an almost double content of total COPs (3.34 µM, approximately 1337 ng /g of chocolate) than the first one (1.69 µM, approximately 675 ng /g of chocolate). Importantly, even in the CaCo-2 cell monolayers treated with six-months stored mixture of COPs oxy-Mix2, no alterations were observed of those cell junctions markedly affected by identical concentration of 7βOHC or 7KC used alone. The junctions’ derangement started to be significantly evident when oxy-Mix2 was used at higher concentration (5 µM, approximately 2 µg oxysterols/g of product) or when treatments were carried out with repeated doses of oxy-Mix2 every 24 hours. Although achieved in a still widely adopted <em>in vitro</em> model system, these findings could orientate the definition of a safe shelf-life for dairy products, certainly for milk chocolate.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"246 ","pages":"Article 106648"},"PeriodicalIF":2.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142755806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}