Search Results (706)

In this study, we review the use of cyclodextrin-based formulations to develop oral tablets of cladribine by enhancing its bioavailability and to improve the solubility and stability of retrometabolic chemical delivery systems (CDSs) in general and estredox, a brain-targeting estradiol-CDS, in particular. Cyclodextrins (CDs), cyclic oligosaccharides that can form host–guest inclusion complexes with a variety of molecules, are widely utilized in pharmaceuticals to increase drug solubility, stability, bioavailability, etc. The stability of the complex depends on how well the guest fits within the cavity of the CD host; a model connecting this to the size of the guest molecules is briefly discussed. Modified CDs, and particularly 2-hydroxypropyl-β-cyclodextrin (HPβCD), provided dramatically increased water solubility and oxidative stability for estredox (estradiol-CDS, E₂-CDS), making its clinical development possible and highlighting the potential of our brain-targeted CDS approach for CNS-targeted delivery with minimal peripheral exposure. A unique HPβCD-based formulation also provided an innovative solution for the development of orally administrable cladribine. The corresponding complex dual CD-complex formed by an amorphous admixture of inclusion- and non-inclusion cladribine–HPβCD complexes led to the development of tablets that provide adequate oral bioavailability for cladribine, as demonstrated in both preclinical and clinical studies. Cladribine–HPβCD tablets (Mavenclad) offer a convenient, effective, and well-tolerated oral therapy for multiple sclerosis, achieving worldwide approval and significant clinical success. Overall, the developments summarized here underscore the importance of tailored cyclodextrin-based approaches for overcoming barriers in drug formulation for compounds with challenging physicochemical properties, and demonstrate the versatility and clinical impact of CD inclusion complexes in modern pharmaceutical development. Full article

Reproductive efficiency is a key determinant of sheep productivity, yet Hetian sheep remain limited by relatively low fecundity despite their adaptability to harsh environments. The inhibitor of DNA binding 2 (ID2) gene is known to regulate cell proliferation and differentiation, but its specific role in sheep reproduction is not well understood. This study aimed to characterize the ID2 gene in Hetian sheep and to assess its functional association with ovarian granulosa cells and litter size. The coding sequence of ovine ID2 was cloned and analyzed using bioinformatics tools. Tissue-specific expression patterns were measured by quantitative PCR at different pubertal stages. A total of 157 ewes were genotyped to identify single-nucleotide polymorphisms (SNPs) and their association with litter size. Functional studies were performed by lentiviral overexpression of ID2 in granulosa cells, with effects evaluated using CCK-8 proliferation assays, ELISA for hormone secretion, and RT-qPCR for related gene expression. ID2 was highly expressed in the ovary, particularly during puberty. Four SNPs (g.18202368 A>T, g.18202372 G>A, g.18202431 G>C, g.18202472 G>C) were significantly associated with increased litter size. Overexpression of ID2 promoted granulosa cell proliferation, increased progesterone, decreased estradiol, and altered expression of key genes in the TGF-β/BMP-SMAD signaling pathway. The ID2 gene plays a crucial role in ovarian function and reproductive regulation in Hetian sheep. Its polymorphisms and functional impact on granulosa cells suggest that ID2 is a promising candidate gene for marker-assisted selection to improve reproductive efficiency in sheep. Full article

Background: Colorectal cancer (CRC) is more prevalent in men, and premenopausal women have a better prognosis than both men and postmenopausal women, suggesting a protective effect of estrogen. Humans are exposed to estrogen-like contaminants such as bisphenol A (BPA), a chemical used in the production of plastics that has been linked to hormone-related malignancies (e.g., breast, ovarian, and prostate cancers). The natural flavonolignan compound silibinin (SIL), acting as an estrogen agonist, may play a protective role in CRC in one or both sexes. Objectives: To explore the possible association between BPA and CRC, focusing on its potential pro-tumor role and possible gender differences. Analyzing the possible protective effects of SIL on the development of CRC is the secondary objective of the project. Methods: To shed light on the interaction between sex and estrogens, both endogenous and exogenous, in the onset of CRC. To this end, we combined ex vivo, in vitro, and in vivo approaches to deepen our understanding of the molecular mechanisms involved. Conclusions: The data provided by this study will contribute to understanding the role of estrogens and their receptors in the onset and progression of CRC and the potential protective role of SIL in both sexes. Full article

Cancer development in BRCA1 carriers is a multi-step process, which is triggered by several factors and mechanisms that are not clearly understood. Most BRCA1 carriers develop triple-negative breast cancer (TNBC)—estrogen receptor (ER)-, progesterone receptor (PR)-, and HER2 -negative cancers—which originates from ER/PR/HER2-negative breast progenitor cells. Due to a lack of ER/PR/HER2-negative cell models with BRCA mutations, the processes inducing cancer development in BRCA carriers have not been comprehensively studied. Thus, studies characterizing ER/PR/HER2-negative cells carrying a BRCA1 germline mutation are needed to gain more in-depth knowledge about the steps leading to cancer initiation in BRCA1 carriers. To study the cancer development in these patients, we established a protocol for the generation of human ER/PR/HER2-negative breast organoids carrying a BRCA1 germline mutation. We confirmed that these organoids are unresponsive to estrogen, can self-renew, and express the stem/progenitor marker CD44. In addition, we observed that these organoids contain outgrowths that resemble the mature ductal and lobular units of the mammary gland, thus making it a suitable model system to study how cancer develops in ER/PR/HER2-negative mammary cells that carry a BRCA1 germline mutation. Full article

Maternal tobacco smoke exposure is associated with impaired fetal growth and long-term disease risk (DOHaD, Developmental Origins of Health and Disease). Whether placental steroid hormones are independently altered remains a matter of debate. We quantified six placental steroids (estradiol, estriol, estrone, progesterone, testosterone, and pregnanediol) using HPLC–Corona CAD in 70 deliveries (C = 30; PS = 20; AS = 20). Distributional differences were assessed with Kruskal–Wallis and pairwise Mann–Whitney tests with Benjamini–Hochberg (BH) control. Adjusted associations used log-linear OLS with HC3 robust SE: Model A (gestational age, maternal BMI, newborn sex) and Model B (Model A + birth weight), reported as percent change vs. controls, computed as (exp(β) − 1) × 100 with 95% CI. Secondary analyses tested (i) multiclass logistic classification of C/PS/AS from the steroid panel (5-fold stratified CV) and (ii) prediction of birth weight (OLS and 2-component PLS). All six steroids differed by group (BH-adjusted p ranging from 9.18 × 10⁻¹² to 6.66 × 10⁻⁸). In Model A, AS vs. C showed lower estrogens/progestins (estradiol, −46.2%; estriol, −24.7%; estrone, −25.9%; progesterone, −28.2%; pregnanediol, −31.4%) and higher testosterone (+40.8%); these effects persisted in Model B after adjusting for birth weight. The panel classified C/PS/AS with 0.900 cross-validated accuracy (weighted OvR AUC 0.994). Hormones poorly predicted birth weight (PLS CV R² = −0.777). Maternal active and passive smoking is associated with a coherent and independent disruption of placental steroidogenesis. A targeted placental steroid panel offers biologically meaningful early markers relevant to DOHaD. Full article

Background/Objectives: Tubo-ovarian high-grade serous carcinoma (HGSC) is a highly lethal malignancy, usually diagnosed at an advanced stage due to the lack of early symptoms and biomarkers. Contraceptive hormone use is associated with a reduced risk of HGSC, but the relative contributions of natural versus synthetic progestins, and their interaction with estrogens, are poorly understood. Methods: We evaluated the chemo-preventive efficacy of a synthetic progestin medroxyprogesterone acetate (MPA), progesterone (P4), and combined 17β-estradiol-progesterone (E2 + P4) in a well-characterized genetically engineered mouse model (GEMM) of oviductal HGSC based on the conditional inactivation of one or both alleles of the Brca1, Trp53, Rb1, and Nf1 tumor suppressor genes (BPRN-het and BPRN-homo mice, respectively). Mice received hormones or placebo via slow-release pellets implanted subcutaneously. After induction of tumor formation, the mice were monitored for tumor development, progression, and survival. Tumor incidence was assessed histologically, and hormone effects were further explored via RNA-seq analysis of oviductal tissues. Results: MPA significantly reduced HGSC incidence and delayed tumor progression compared to the placebo, P4, and P4 + E2 in both BPRN-homo and BPRN-het mice, with up to 78% tumor-free survival in the MPA-treated BPRN-het cohort. P4 monotherapy did not provide significant protection vs. the placebo, but the effects of P4 could have been impacted by a failure to achieve sustained release of the hormone beyond 4–8 weeks. The E2 + P4 combination accelerated tumorigenesis and reduced survival (p < 0.0001 in BPRN-homo and p = 0.0004 in BPRN-het mice). MPA did not affect tumorigenesis in a colon cancer GEMM, or the growth of mouse HGSC-derived cells in vivo, suggesting the role of MPA in the early stages of HGSC development. Gene expression analyses showed that P4 and MPA downregulated cholesterol homeostasis, early and late estrogen response, and epithelial–mesenchymal transition pathways, though only MPA afforded tumor protection. Conclusions: These findings demonstrate that a synthetic progestin, specifically MPA, confers robust protection against HGSC development, while a combination including E2 (E2 + P4) increases risk. This work also illustrates how HGSC GEMMs can be used to compare the chemo-preventive effects of various synthetic progestins on HGSC development in order to prioritize the most effective ones for use in preventing HGSC in both general and high-risk populations. Full article

Background/Objectives: Oxidative stress is a critical factor in the development and progression of endometriosis. Granulosa cells, which reside near oocytes in follicles, exhibit steroidogenic activity, and, consequently, influence oocyte quality. Increased oxidative stress may induce the danger signal such as HMGB-1 in granulosa cells and eventually change the follicular environment of patients with endometriosis. This study aimed to demonstrate that HMGB-1 and its receptors, TLR4 and RAGE, play important roles in the changes in the follicular environment in infertile patients with endometriosis. Methods: In the immortalized human granulosa cell line (hGL5), cell proliferation and apoptosis assay, ELISA for estradiol, qRT-PCR for HMGB-1 and TLR4, Western blot for apoptosis-related and NF-κB pathway-related proteins, and ELISA for inflammatory molecules IL-1β and IL-6 were performed after H₂O₂ treatment. Results: H₂O₂ treatment to the hGL5 cell line decreased cell proliferation via apoptosis and, as a result, decreased steroidogenesis. Also, it increased the gene expression of HMGB-1 and TLR4, increased the protein expression related to the NF-κB pathway, and increased the release of inflammatory molecules IL-1β and IL-6. Conclusions: The results indicate that oxidative stress associated with endometriosis may increase inflammation by interacting with HMGB-1 and TLR4 and activating the NF-κB pathway to increase proinflammatory responses. The findings of this study may provide insight into endometriosis with decreased oocyte quality. Full article

The present study investigated the effects of fermented Chinese herbal (FCH) compounds on the egg production, egg shell quality, and egg yolk cholesterol of laying hens. A total of 1260 Hy-Line pink laying hens, 34 weeks old, were randomly divided into three groups, with six replicates per group and 70 hens per replicate, as follows: the control group (CON group) was fed a diet without FCH compounds, and the 2% FCH group and the 3% FCH group were fed a diet supplemented with 2% FCH and 3% FCH, respectively. The results show that the FCH compound significantly increased the laying rate compared to the CON group (p < 0.05). Analyses of the serum biochemical indices showed that supplementation with FCH compound significantly decreased the levels of total cholesterol (TC), total triglyceride (TG), high-density lipoprotein cholesterol (HLDL-c), low-density lipoprotein cholesterol (LDL-c), very-low-density lipoprotein cholesterol (VLVL-c), aspartate transaminase (AST), and alanine aminotransferase (ALT) (p < 0.05) and increased the serum total bile acids, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and 17-β-Estradiol (E2) levels (p < 0.05). The FCH group significantly increased the activity of superoxide dismutase (SOD) and total antioxidant capacity and decreased malondialdehyde (MDA) levels in the liver and uterus compared to the CON (p < 0.05). FCH supplementation was also associated with improved egg quality, seen through factors including enhanced yolk color, albumen height, Haugh unit score, eggshell strength, and thickness and reduced egg breaking rate and TC and TG contents in egg yolk. The gene expression analyses showed that FCH supplementation significantly increased the calcium metabolism-related gene expression (CaBP-D_28k, NCX, VDR, CYP₂₇B₁, OPN, PMCA, CA₂) in duodenum, kidney, and uterus tissues compared to the CON group (p < 0.05). FCH significantly repressed FAS and HMGCR mRNA expression and enhanced CYP7A1 mRNA expression in the liver (p < 0.05). These results indicate that diet supplementation with FCH compounds may improve egg quality by regulating reproductive hormones, lipid metabolism, and calcium metabolism. Full article

Background/Objectives: The Sox transcription factor family is critical for gonadal development and sex differentiation in animals, yet its roles in chelonians, particularly in the Chinese soft-shelled turtle (Pelodiscus sinensis), have rarely been investigated. Methods: This study cloned and analyzed the cDNA sequences of Sox3 and Sox30 genes from P. sinensis, examining their amino acid sequences and structural properties. Real-time quantitative PCR (RT-qPCR) was used to assess the expression of these two genes in different adult tissues and at various stages of embryonic gonadal development. Additionally, the effects of exogenous hormones (17β-estradiol, E₂ and 17α-Methyltestosterone, MT) on the expression of Sox3 and Sox30 were also investigated. Results: The results indicated that Sox3 showed significantly elevated expression in female gonads, kidney, brain, liver, lung, spleen, and muscle relative to male counterparts, displaying a female-biased expression pattern. In contrast, Sox30 showed a male-biased pattern, with higher expression in male gonads, spleen, muscle, brain, and liver than in females, showing expression. Both genes were expressed at low levels. Exogenous hormone treatments revealed that MT significantly downregulated Sox3 expression in female embryos, whereas E₂ significantly enhanced Sox3 expression in male embryos. Furthermore, MT treatment significantly upregulated Sox30 expression in female embryos, and E₂ treatment also significantly increased Sox30 expression in male embryos. Conclusions: These findings suggest that Sox3 and Sox30 play crucial roles in the gonadal development of P. sinensis, with Sox3 potentially involved in ovarian development and Sox30 in testicular maturation. Both genes are regulated by exogenous hormones, highlighting their importance in sex differentiation and gonadal development. This study provides valuable theoretical insights for further exploration of the molecular mechanisms of sex regulation in reptiles. Full article

Daidzein and genistein are abundant in soy-rich foods, whose supplementation has been proposed to have beneficial effects on several diseases, including diabetes mellitus and obesity. 17β-estradiol (E2) enhances the expression of the Slc2a4 gene and GLUT4 protein in adipose tissue, increasing glucose consumption and contributing to glycemic control. We investigated, in 3T3-L1 adipocytes, the effect of low and high doses of daidzein and genistein on Slc2a4/GLUT4 expression and the participation of estrogen receptors 1/2 (ESR1/ESR2) in the regulations observed. Differentiated adipocytes were cultivated, for 24 h, in the presence of 17β-estradiol (E2, 10 nM), daidzein (10 nM–150 μM) and genistein (10 nM–50 μM), with or without ESR1 or ESR2 antagonists. Daidzein/genistein at a low dose (10 nM) increased Slc2a4/GLUT4 expression (50%, p < 0.05), an effect abrogated by an ESR1 antagonist, mimicking the effect of E2. However, maximal doses of daidzein and genistein reduced, in a ESR1-mediated mechanism, the expression of mRNA (by 47% and 60%, p < 0.001) and the protein (by 29% and 36%, p < 0.01), respectively, for daidzein and genistein, as compared to E2. In conclusion, in adipocytes, daidzein and genistein have a biphasic ESR1-mediated effect: while low concentrations increase the expression of Slc2a4/GLUT4, high concentrations decrease it, the former predisposing to an adipogenic effect, the latter to a diabetogenic condition. Full article

Hormone replacement therapy (HRT) currently represents the first-line treatment to manage and reduce menopausal symptoms. Standard regimens generally combine 17β-estradiol (E2) or conjugated equine estrogens (CEEs) with micronized progesterone (P4) or synthetic progestins. While synthetic progestins ensure endometrial protection against estrogen-induced stimulation of the endometrium, their impact on metabolic, cardiovascular, skeletal, and cognitive systems is heterogeneous and not always beneficial. In contrast, progesterone, as a micronized preparation (P4), allows for more physiological effects because it is chemically identical to endogenous progesterone. This narrative review provides an updated overview of the clinical benefits of HRT regimens based on E2/P4, with a focus on their impact on endometrial thickness, venous thromboembolism (VTE), cardiovascular diseases (CVDs), breast cancer risk, cognitive effects, bone protection, and quality of life (QoL). Full article

Skeletal muscle homeostasis is dependent on the satellite cell pool, which is regulated by numerous signaling pathways. Estradiol (E2) function via estrogen receptor alpha (ERα, Esr1) plays an important role in satellite cell regulation in females, being necessary for satellite cell maintenance, proliferation and differentiation. Here we investigate this signaling axis in male satellite cells. Male satellite cells express Esr1 mRNA at similar levels to female satellite cells, and E2 enhances the proliferation of male satellite cell-derived myoblasts in vitro. Deletion of Esr1 specifically in male satellite cells has no effect on satellite cell number, nor on their ability to self-renew after injury, during regeneration, or when transplanted into male hosts. However, Esr1 deletion severely reduces self-renewal of male satellite cells when transplanted into female hosts. These data suggest that male satellite cells are competent for E2-ERα signaling, but that this signaling is not efficacious in the male environment, though E2-ERα signaling does become necessary when the male cells are transplanted into a female environment. Full article

17β-estradiol (E2) enhances the cerebellar molecular layer interneurons (MLIs)—Purkinje cells (PCs) synaptic transmission via activation of the Erβ in vivo in mice. Whether E2 regulates cerebellar MLI-PC synaptic plasticity is unknown. To investigate the mechanism of E2, we evaluated the modulation of facial stimulation-evoked MLI-PC long-term plasticity in mice. Cell-attached recordings from PCs of Crus II were performed using an Axopatch-700B patch-clamp amplifier. The MLI-PC synaptic transmission was evoked by facial stimulation. Immunohistochemistry was used to detect the expression of ERβ. Under control conditions, 1 Hz facial stimuli induced long-term depression (LTD) at MLI-PC synapses, characterized by a sustained reduction in P1 amplitude and a simple spike (SS) pause. The facial stimulus-induced MLI-PC LTD was completely prevented by E2, but this effect was reversed by a selective ERα/ERβ antagonist, ICI182780. Blockade of cannabinoid receptor 1 (CB1R) eliminated the MLI-PC LTD under control conditions, but revealed an E2-triggered long-term potentiation (LTP). The E2-triggered MLI-PC LTP persisted in the presence of an ERα antagonist but was absent in the presence of an ERβ antagonist PHTPP. The E2-triggered MLI-PC LTP remained unaffected by protein kinase A inhibition but was abolished by inhibition of protein kinase C (PKC) and intracellular Ca²⁺ depletion. Moreover, ERβ immunoreactivity was abundantly distributed around dendrites and somas of PCs in the Crus II region of the mouse cerebellar cortex. The present results suggest that E2 activates ERβ, thereby triggering facial stimulation-induced MLI-PC LTP via the PKC signaling cascade, which occludes CB1R-dependent MLI-PC LTD in the cerebellar cortex of mice in vivo. Full article

To understand the changes in gonadal sex differentiation, digestive enzyme activity, and growth-related hormone levels in the larval and juvenile black scraper, Thamnaconus modestus, continuous sampling was conducted from 0 to 91 days post-hatching (dph). 17β-estradiol (E2) and testosterone (T) levels, six digestive enzymes, as well as T3, T4, GH, and IGF-I were detected. The results showed that oogonia or spermatogonia was observed at 60 dph. During the sex differentiation to female or male, both E2 and T levels significantly increased (p < 0.05), suggesting that E2 and T may induce the sex differentiation to female or male in T. modestus, respectively. The amylase activity from 0 to 35 dph showed a slow upward trend, which may be due to the transition from endogenous to exogenous nutrition at this time. From 12 to 25 dph, alkaline protease activity significantly decreased (p < 0.05), while acid protease levels significantly increased (p < 0.05), suggesting that as organs in the digestive system continue to develop, acid protease plays an important role. T3 and T4 could already be detected at 0 dph, and the T4 content was always much higher than T3 throughout the stages, indicating that T4 may play more important roles than T3. Additionally, the changes in IGF-I and GH content followed a trend of an initial increase, a subsequent decrease, and then an increase, ultimately showing an overall upward trend. These results indicate that T4, IGF-I, and GH play crucial roles in growth and development in the juvenile fish. In conclusion, the results of this study provide useful information for growth, artificial reproduction, and sex regulation in T. modestus. Full article

Breast cancer remains a leading cause of global mortality. According to international cancer data, significant progress has been made in treating breast cancer; however, metastasis and drug resistance continue to be the primary causes of mortality for many patients. This study investigated the modulation of apoptosis-related genes in response to ionizing radiation and estrogen exposure based on a human breast epithelial cell model (MCF-10F and its transformed variants: Estrogen, Alpha3, Alpha5, Tumor2) previously established, where cells were treated with high linear energy transfer alpha particles, with or without 17β-estradiol. Gene expression profiling was performed using an Affymetrix U133A microarray, and bioinformatic analyses assessed differential expression, estrogen receptor status, and correlations with overall survival. Distinct gene expression patterns emerged across cell lines and tumor subtypes. TP53 expression correlated positively with TP63, BIK, CFLAR, BIRC3, and BCLAF1. TP63, PERP, CFLAR, BCLAF1, GULP1, and BIRC3 were elevated in normal tissue, whereas BIK, PHLDA2, and BBC3 were upregulated in tumors. ER-positive tumors exhibited higher TP63, BIK, BCLAF1, and BBC3 expression, while ER-negative tumors showed increased PERP, CFLAR, BIRC3, and PHLDA2. Notably, elevated BCLAF1 expression was associated with poorer survival in Luminal A patients, and high PHLDA2 expression correlated with reduced survival in Luminal B cases. These findings indicate that resistance to apoptosis is a fundamental mechanism in breast cancer progression and therapeutic evasion. Breast tumors selectively alter the expression of key genes to promote growth, evade apoptosis, and develop therapeutic resistance. The differential expression and correlations of these apoptosis-related genes highlight their potential as molecular targets for future personalized cancer therapies and as valuable biomarkers for prognostic stratification and predicting therapeutic response. Full article