Search Results (1,849)

Precise quantitative monitoring of 17β-estradiol (E2) is important for reproductive management in precision livestock farming. However, E2 determination in complex biological matrices remains challenging because of matrix-derived background and signal variability. Here, we developed a nanoconfinement-assisted solid-state ratiometric fluorescent aptasensor integrating target-induced strand displacement (TISD), magnetic separation, and anodic aluminum oxide (AAO) nanochannel confinement. The sensing probe consisted of streptavidin-coated magnetic nanoparticles (MNPs) carrying a FAM-labeled cDNA internal reference and a Texas Red-labeled E2 aptamer reporter. E2 binding promoted dissociation of the Texas Red-labeled aptamer from the magnetic probe. Magnetic separation and washing reduced soluble matrix-derived interference, while subsequent deposition of the sensing complexes onto an AAO membrane mitigated coffee-ring-associated nonuniformity and produced a more spatially uniform dual-color fluorescence distribution for ratiometric analysis. Under matrix-matched calibration conditions, linear ranges of 5.0–50.0 pM were obtained in tap water and sow saliva, 5.0–40.0 pM in whole milk, and 5.0–15.0 pM in post-estrus sow urine. The LOD determined in tap water was 3.62 pM. The different calibration slopes obtained among the four matrices indicated that residual matrix-dependent effects remained and that matrix-specific calibration was required for quantitative analysis. Matrix-matched spike recoveries ranged from 86.92% to 119.54% across the investigated matrices. The aptasensor exhibited the strongest response toward 17β-E2 among the tested compounds; however, cross-reactivities of 77.3% for E3 and 47.3% for 17α-E2 indicated preferential rather than exclusive recognition. Molecular docking suggested a putative binding pose but did not experimentally establish the molecular recognition mechanism. Overall, the platform demonstrated laboratory-scale analytical feasibility in pretreated tap water, sow saliva, whole milk, and post-estrus sow urine. Further development of sample preparation, magnetic handling, membrane loading, probe selectivity, and portable fluorescence readout will be required before in situ or on-site application. Full article

Optimizing farrowing management is crucial in swine production, yet the combined effects of induction protocols and stimulants on the sow remain poorly understood. This study evaluated the effects of farrowing induction using cloprostenol as a single dose (full dose) or split dose, with or without caffeine administration, on the performance, behavior, and hormonal concentrations of primiparous (n = 25) and multiparous (n = 25) sows. Sows were assigned to five treatments: Control (n = 10), PGF2α Full Dose (175 μg) (n = 10), PGF2α Split Dose (87.5 + 87.5 μg) (n = 10), PGF2α Full Dose + Caffeine (420 mg) (n = 10), and PGF2α Split Dose + Caffeine (420 mg) (n = 10). The PGF2α Split Dose and PGF2α Split Dose + Caffeine treatments significantly reduced farrowing duration by up to 100 min in primiparous sows (122.40 ± 82.24 and 136.00 ± 38.33 min, respectively; p = 0.002), and in multiparous sows by up to 76 min (134.80 ± 73.84 and 158.00 ± 44.82 min, respectively; p < 0.05), compared to the Control and PGF2α Full Dose groups. Furthermore, the PGF2α Split Dose induction method lowered the frequency of stress-related behaviors (0.01 and 0.08 for PGF2α SD, and 0.03 and 0.08 for PGF2α SD + Caffeine, for gilts and sows respectively), with sows showing fewer posture changes and maintaining more stable resting positions, particularly the lying lateral posture (0.86 for PGF2α SD and 0.96 for PGF2α SD + Caffeine). Cortisol levels varied by parity, with multiparous sows showing higher prepartum and intrapartum levels, while primiparous sows peaked twelve hours postpartum; estradiol levels remained unaffected by treatments. In conclusion, inducing farrowing with a split-dose prostaglandin protocol is highly efficient, and the addition of caffeine further enhances farrowing performance while significantly improving behavioral welfare by reducing standing postures associated with discomfort in both primiparous and multiparous sows. Full article

Mentha pulegium L. is reported to contain phytochemicals known to bind to estrogen receptors and modulate estrogenic effects. A hydroethanolic leaf extract of Mentha pulegium L. (MPE) was prepared, and its effects on uterine and ovarian tissues in immature rats were investigated, focusing on transcriptional endpoint-related estrogenic activity. Female Sprague Dawley rats were treated with varying doses of MPE alone or in combination with estradiol for seven days. Gene expression analysis was performed using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to evaluate the effect of MPE on estrogen-responsive biomarkers: Calbindin-D9k (CaBP-9k), Progesterone receptor (Pgr), Trefoil factor 1 (pS2), Intestinal calcium-binding protein integral (Icabp), Integral membrane-associated protein-1 (Itmap1) and Complement component 3 (CC3) genes. In ovarian tissues, MPE treatment decreased CaBP-9k and Icapb expression, with CC3 showing significant decreases in the 200 mg/kg group. Treatments with MPE and estradiol significantly reduced the expression of all estrogen-responsive genes compared to estradiol treatment. In uterine tissues, 1000 mg/kg MPE increased CaBP-9k and pS2 expression significantly but decreased Icapb, CC3, pS2, and Itmap across all treatment groups significantly. Combined estradiol treatment with MPE (500 and 1000) mg/kg showed significantly low CaBP-9k and CC3 expressions. Increased expression of Icapb, Itmap, and pS2 was observed when combined estradiol treatments with MPE (500 and 1000) mg/kg were compared to estradiol treatment. MPE influenced the expression of specific genes in the uterus and ovaries and thus may exhibit endocrine-modulatory activity by multiple mechanisms of action, highlighting its potential complexity in modulating estrogenic responses. Full article

This study aimed to determine the action of a single dose of human chorionic gonadotropin (hCG) on follicular dynamics, CL functionality, ovarian (OV.A) and uterine arteries (MUA) blood flow, hormones (estradiol [E2] and luteinizing hormone [LH]), nitric oxide (NO), and vascular endothelial growth factor (VEGF) levels in Thoroughbred mares. Animals were subjected to ultrasonography scanning, when the dominant follicle (F1) reached 35 mm in diameter, mares were injected with hCG (hCG group; n = 7) at hour 0, while others were injected with saline (control; n = 7). The examination was performed a those points at hours −24 h, −12 h, 0 h, 12 h, 24 h, 36 h, 48 h, 72 h, and 96 h. Blood sampling, ultrasound scanning, hormone assaying, and statistical analysis were performed. F1 was ruptured at 12 h after injection compared to 24 h in the control, with an elevation in its diameter (p ≤ 0.05) and a colored area surrounding F1. The amount of color Doppler area around CL and its diameter were increased (p ≤ 0.05) at 24 and 48 h after injection with an increase (p ≤ 0.05) in the ipsilateral uterine horn colored area from 6 to 36 h after injection. OV.A and MUA Doppler velocity parameters were elevated (p ≤ 0.05) at 24 h. Both NO and VEGF levels were elevated at 24 h after injection, with the elevation (p ≤ 0.05) of E2 occurring only before injection and then declining. hCG and LH levels were increased after injection at 12 and 24 h, respectively, with an interaction (p ≤ 0.05) effect. hCG may speed up and hasten the ovulation process by elevating follicular color amount and CL morphometry via elevating ovarian flow and uterine body total coloration via elevating uterine blood flow (declining Doppler indices) in mares. Full article

Polypropylene microplastics (PP-MPs) are ubiquitous in our daily lives, but their toxicological effects on mammals remain poorly understood. This study investigated the toxicity effects of PP-MPs on C57BL/6 mice using 16S rRNA gene amplicon sequencing and transcriptome sequencing. Female and male mice were randomly classified into the control and PP-MPs-treated groups, respectively, and the experiment lasted for 5 weeks. We found that PP-MPs exposure did not affect the levels of immunoglobulin G (IgG), interleukin-4 (IL-4), and interferon gamma (IFN-γ), indicating that humoral immunity and inflammatory levels were not influenced by PP-MPs treatment. However, PP-MPs exposure reduced the PHA response in female mice, but not in male mice. It also did not alter the wet mass of testicles and ovaries, nor the levels of testosterone and estradiol. Exposure to PP-MPs altered the expression of the testicular genes. G protein-coupled receptor signaling pathways, olfactory receptor activity, and protein digestion and absorption were downregulated in the PP group. Collagen genes (Col9a3, Col11a2, Col27a1, Col26a1, Col7a1) play a significant role in downregulating protein digestion and absorption. In addition, PP-MPs exposure caused a change in beta diversity of gut microbiota, indicating the alteration of their community structure. PP-MPs exposure reduced the relative abundance of the probiotic Lactobacillus. Changes in the gut microbiota may be related to the expression levels of testicular genes. Overall, PP-MPs exposure altered both the community structure of the gut microbiota and the expression levels of testicular genes in mice, and collagen genes may serve as a critical factor influencing testicular function. Full article

We explored the potential functions of differentially expressed miRNAs, mRNAs, lncRNAs, and circRNAs identified in the ovaries of gilts with normal estrus (NE) and follicular cyst-associated anestrus (AE), as well as their putative competing endogenous RNA (ceRNA) regulatory networks. Ovarian morphology was assessed via ultrasonography, and serum concentrations of Follicle-stimulating hormone (FSH), (Estradiol) E2, and Progesterone(P4) were measured. Ovarian tissues were harvested after slaughter for whole-transcriptome sequencing. Bioinformatic tools were used to screen differentially expressed RNAs(DERNAs) between NE and AE gilts. We further predicted target interactions among these transcripts, conducted functional enrichment analysis on target genes, and constructed candidate ceRNA regulatory networks potentially associated with gilt estrus. Phenotypic verification confirmed that ovarian ultrasonographic characteristics, histological morphology, and serum reproductive hormone levels were consistent with the physiological status of NE and AE gilts. Under the screening thresholds of p < 0.05 and |log₂FC| ≥ 1, we identified 22 lncRNAs that may interact with 21 mRNAs via 50 miRNAs, alongside 39 circRNAs predicted to regulate 26 mRNAs through 72 miRNAs. Functional enrichment analysis indicated that target genes of these differentially expressed transcripts were predominantly enriched in the lysosome pathway, PPAR signaling pathway, chemokine signaling pathway, cholesterol metabolism and NOD-like receptor signaling pathway. Hub molecules including FGF1, GHR, TLR2, ssc-miR-370, and miR-21-5p were shared in lncRNA/circRNA-miRNA-mRNA regulatory networks; these molecules have been reported to participate in progesterone synthesis, estrus modulation, and endocrine homeostasis. Of particular interest, two non-coding RNAs, MSTRG.1285.1 and novel_circ_056113, were predicted to act as candidate ceRNAs that may sponge ssc-miR-370 and miR-21-5p, which could in turn modulate the expression of estrus-associated mRNAs including FGF1, GHR, and TLR2. The expression trends of MSTRG.1285.1, novel_circ_056113, miR-370, miR-21-5p, GHR, TLR2, and FGF1 were validated by qRT-PCR, and the quantification results agreed with transcriptome sequencing data. Collectively, this study constructed a predicted ceRNA regulatory network of ovarian transcripts comparing NE and AE gilts and uncovered multiple RNA molecules potentially involved in estrus regulation. These findings offer preliminary theoretical clues for exploring the onset of puberty in gilts. Full article

Background/Objectives: Isorhamnetin is a naturally occurring flavonoid with reported antioxidant, anti-inflammatory, and anticancer properties. Although its biological activities have been extensively investigated, its effects on ovarian cell physiology remain insufficiently characterized. This study aimed to evaluate the influence of isorhamnetin on the viability, steroid hormone secretion, apoptosis, and growth factor signalling in human ovarian cell lines representing both non-tumour and tumour phenotypes. Methods: Human granulosa (HGL5), granulosa tumour (COV434), and epithelial ovarian carcinoma (OVCAR-3) cell lines were treated with isorhamnetin at concentrations of 5–80 μg/mL for 24 h. Cell viability was determined using the AlamarBlue^TM assay. The secretion of progesterone, 17β-estradiol, and the presence of transforming growth factor β2 (TGF-β2), transforming growth factor β receptor 2 (TGFBR2), and apoptosis-inducing factor (AIF) was quantified by enzyme-linked immunosorbent assay (ELISA). Results: Isorhamnetin significantly reduced cell viability in a dose-dependent manner, with tumour cell lines exhibiting greater sensitivity than non-tumour granulosa cells. A significant decrease in viability was observed in OVCAR-3 cells from 10 μg/mL onward, whereas COV434 cells showed significant reductions at concentrations of 20 μg/mL and higher. In contrast, HGL5 cell viability was significantly affected only at the highest concentration (80 μg/mL). No significant changes were detected in the secretion of progesterone, 17β-estradiol, TGF-β2, or TGFBR2 in any of the examined cell lines. A significant reduction in AIF production was observed only in HGL5 cells treated with 80 μg/mL isorhamnetin. Conclusions: Isorhamnetin preferentially decreased the viability of ovarian tumour cells while exerting only limited effects on non-tumour granulosa cells, indicating preferential cytotoxic activity toward malignant ovarian cells. The absence of significant changes in steroid hormone production and TGF-β suggests that its antiproliferative effects involve molecular pathways that were not investigated in the present study. These findings support further investigation of isorhamnetin as a potential natural compound for ovarian cancer prevention or adjunctive therapy. Full article

Estrogens, particularly the most active 17β-estradiol (E2), play a crucial role in the initiation and development of breast cancer. Therefore, current treatment strategies are based on interfering with estrogen receptors (ERs) via pure antiestrogens or selective modulators (SERMs) or by inhibiting estrogen synthesis through key enzymes in this pathway. While SERMs are generally effective, resistance often develops. Moreover, they cannot be used to treat triple-negative breast cancers (TNBCs). Three critical enzymes in the estrogen synthesis pathway, namely aromatase, sulfatase, and 17β-hydroxysteroid dehydrogenases, are of particular interest as drug targets. Some, such as aromatase inhibitors, are already used in clinical practice, while the other two remain under intensive investigation. Several comprehensive reviews have been published on ER modulators or individual enzyme inhibitors. This paper attempts to summarize recent data on modulators/inhibitors of both the ER and the major enzymes involved in estrogen biosynthesis and offers a perspective on their further development. Full article

While dihydrotestosterone (DHT) has traditionally been considered the principal steroid in benign prostatic hyperplasia (BPH) pathogenesis, the role of estradiol (E2) and its relationship with local androgen accumulation remain unclear. This study investigated whether transition zone (TZ) tissue concentrations of E2, testosterone, and DHT are associated with prostate enlargement. TZ tissue cores were collected from 80 men undergoing prostate biopsy. Patients were stratified by total prostate volume (TPV): small (<30 mL) and enlarged (≥30 mL) prostate groups. Steroids were quantified using liquid chromatography-high resolution mass spectrometry. Tissue concentrations of E2, testosterone, and DHT were significantly higher in enlarged prostates (E2: 0.031 vs. 0.012 ng/g; testosterone: 0.804 vs. 0.517 ng/g; DHT: 7.903 vs. 3.526 ng/g; all p < 0.05). Across the cohort, TPV strongly correlated with DHT (R = 0.870, p < 0.001), E2 (R = 0.845, p < 0.001), and testosterone (R = 0.817, p < 0.001). These findings demonstrate a strong association between local steroid accumulation and prostate enlargement, supporting the hypothesis that coordinated androgenic and estrogenic activity shapes the hormonal microenvironment driving BPH progression. This pattern aligns with a model of local steroidal hyperfunction that may amplify stromal–epithelial signaling in the enlarged prostate. Full article

Objective: The study was conducted to investigate the frequency of MED12 gene (rs5030619) mutations and to evaluate DNA methylation patterns in uterine leiomyomas among women at a tertiary care hospital in Karachi, Pakistan. Methods: In this cross-sectional study, 200 women with uterine fibroids and 50 controls were recruited from tertiary care hospitals after ethical approval. Baseline data and history were collected. Serum levels of estradiol, progesterone, and FSH were measured. Genomic DNA was extracted from fibroid and myometrial tissues and analyzed using allele-specific PCR for MED12 genotyping. DNA methylation profiling was conducted to evaluate epigenetic modifications. Statistical analyses were performed using the Statistical Package for social sciences (SPSS) version 27.0. Results: The majority of cases were aged 31–50 years, with 41.5% overweight and 24.5% obese. An increased WHR was observed in 65% of cases, compared to only 36% of controls. Serum estradiol, progesterone, and FSH levels were significantly higher in cases across all menstrual phases (p < 0.01). The MED12 A/C genotype was significantly associated with fibroid risk (OR = 11.72, 95% CI: 5.59–24.57, p < 0.001), and the C/C genotype conferred the highest risk (OR = 18.17, 95% CI: 4.00–82.54, p = 0.001). C allele was associated with increased disease susceptibility (OR = 4.65, 95% CI: 2.69–8.03, p < 0.001). Hyper-methylation was the most prevalent finding, observed in 125 cases of leiomyomas (69.4%). Multivariable logistic regression identified age (OR = 1.04, p = 0.012), BMI (OR = 1.09, p = 0.004), multiple fibroids (OR = 2.31, p = 0.005), intramural fibroid location (OR = 1.88, p = 0.041), and positive family history (OR = 2.74, p = 0.003) as significant independent predictors. Conclusions: Uterine leiomyomas may be influenced by a combination of hormonal, metabolic, genetic, and epigenetic factors. Overall findings suggest that MED12 C allele and A/C genotype variation was significantly associated with increased DNA methylation in uterine leiomyomas. Full article

Background: Although genetic-environmental interactions are established in autism spectrum disorder (ASD), how environmental toxicants confer susceptibility remains unclear. This study aimed to investigate potential relationship between genetically predicted cerebrospinal fluid (CSF), metabolite levels and ASD liability, and to prioritize regulatory genes, key pathways, and candidate environmental toxicants. Methods: Using two ASD GWAS datasets (exploration data: 18,381 ASD cases/27,969 controls; validation data: 18,235 ASD cases/36,741 controls), we applied multi-omics approaches to prioritize ASD-associated CSF metabolites, regulatory SNPs, and genes. Enrichment analysis and protein–protein interaction (PPI) network analysis were performed on these metabolite-related genes to explore the potential mechanisms linking CSF metabolic disturbances to ASD. Finally, candidate environmental neurotoxicants were screened through protein-chemical interaction analysis, with binding relationships assessed via molecular docking prediction. Results: Two-sample Mendelian randomization (MR) analysis prioritized adenine and proline as candidate CSF metabolites with potential risk associations with ASD. Summary-data-based MR (SMR) prioritized 39 brain-specific quantitative trait loci (QTL) involving 35 candidate regulatory genes, including dual-metabolite modulator GRM8. Functional enrichment analyses suggested potential associations with mitochondrial dysfunction, Hippo signaling pathway, and microtubule dynamics impairment, with protein–protein interaction networks highlighting KATNA1/KATNAL2 as hubs. Protein-chemical interaction screening nominated 14 candidate environmental toxicants, including established chemicals (acetaminophen, valproic acid, estradiol) and novel candidates (SB-431542, K 7174, benzo[a]pyrene), with docking affinity assessed computationally. Conclusions: Our study provides suggestive evidence that elevated adenine and proline may be potential risk factors for ASD and suggests possible involvement of the mitochondrial–Hippo–microtubule pathway. We also propose benzo[a]pyrene as a candidate environmental toxicant that may perturb CSF metabolism. However, given the limited statistical significance, these findings require further validation. Full article

Breast cancer (BC) is the most incidental cancer in women. Patients’ living conditions and survival rates have significantly improved with the success of two first-line endocrine therapies of selective estrogen-receptor modulators (SERMs from 1977) and aromatase inhibitors (AIs from 1995). Unfortunately, both therapies have produced significant resistance. AI resistance reaches 50% in metastatic estrogen-dependent BC cases, leading oncologists to seek a substitute for current therapies. We target the reductive 17β-hydroxysteroid dehydrogenase type 7 (17β-HSD7), which stimulates the synthesis of the active estrogen estradiol (E2) and reduces the potent androgen dihydrotestosterone (DHT) simultaneously, triggering negative feedback on 17β-HSD7 expression. INH7(464), an improved 17β-HSD7 inhibitor, efficiently blocks estrogen conversion at an IC₅₀ of 92 ± 8 nM. INH7(464) inhibited cell proliferation by decreasing E2 levels and restoring DHT, successively arresting the cell cycle in G0/G1 phase. In vivo, INH7(464) reduced tumor size by 49% on Day 22 in BC xenografts with T47D. INH7(464) reduced E2 levels in mouse blood circulation and xenograft tumor tissue. There is no demonstrated cytotoxicity or cardiotoxicity of INH7(464) on cellular levels and mouse models under the experimental conditions. 17β-HSD7 is a potential target for estrogen-dependent BC therapy, while INH7(464) is promising for a preclinical lead compound (PCT/CA2022/050966). Full article

The reproductive and developmental toxicity of diquat (DQ), a widely used herbicide, during pregnancy remains insufficiently characterized. This study investigated the effects of repeated DQ exposure on fetal development and maternal metabolism in pregnant Sprague–Dawley rats. The rats were administered sublethal doses of DQ (28.88 and 57.57 mg/kg), and maternal serum sex hormones, fetal skeletal development, and serum metabolomic profiles were evaluated. Compared with controls, serum estradiol levels were significantly decreased in the high-dose exposure group, whereas luteinizing hormone and follicle-stimulating hormone levels remained unchanged. Significant alterations in malondialdehyde, superoxide dismutase, and glutathione levels indicated oxidative stress in exposed rats. Severe fetal skeletal ossification deficits and developmental delays were observed, with incomplete or failed ossification of the occipital bone, sternum, caudal vertebrae, and pelvis, with litter-level incidences reaching 100%. Metabolomic analysis identified distinct serum metabolic profiles between DQ-exposed and control groups, suggesting ovarian steroidogenesis, phenylalanine metabolism, and aldosterone synthesis and secretion as key pathways affected by DQ toxicity. These findings suggest that repeated maternal exposure to DQ induces oxidative stress, disrupts metabolic and hormonal homeostasis, and causes delayed fetal skeletal development. This study provides novel evidence for understanding the reproductive and developmental toxicity of DQ and its potential metabolic mechanisms. Full article

Background: Sexual function in postmenopausal women is influenced by both hormonal and psychological factors. This study investigates the associations between sex hormones, sexual function, and sexual beliefs in this population. Objective: This research seeks to assess the relationship between sex hormones, sexual function, and dysfunctional sexual beliefs in postmenopausal women. Methods: A cross-sectional study was conducted with 42 postmenopausal women aged 45–65 years. Instruments included the FSFI and SDBQ. Hormones assessed were testosterone, estradiol, progesterone, prolactin, DHEA, SHBG, and LH. Blood samples were collected in the morning and analyzed using chemiluminescence or radioimmunoassay. Pearson correlation tests were used, with Bonferroni adjustment applied to control for multiple comparisons. Results: Free testosterone was positively correlated with sexual desire and negatively associated with dysfunctional beliefs regarding sexual desire. Estradiol also showed a positive correlation with desire, while prolactin was negatively associated. No other FSFI domains showed significant hormonal associations. Conclusions: Findings suggest that testosterone may influence both sexual desire and the internalization of dysfunctional sexual beliefs in postmenopausal women, highlighting the interplay between biological and psychological dimensions of sexuality, which may be relevant for a more comprehensive clinical assessment of sexual function in this population. Full article

Neuroglobin (NGB), initially identified for its oxygen-binding capacity in neuronal tissues, has emerged as a multifunctional protein involved in neuroprotection, oxidative stress regulation, and mitochondrial homeostasis. Although its functions have been extensively studied in the central nervous system (CNS), its potential role in the gut–brain axis (GBA) remains largely unexplored. The GBA integrates neural, endocrine, immune, and metabolic signaling between the gastrointestinal tract and the brain, and growing evidence from microbiome, neurobiology, endocrinology, and nutrition research suggests that several pathways involved in GBA signaling may also influence NGB expression or activity. However, direct experimental evidence, particularly from in vivo studies, remains limited. This hypothesis-driven review integrates evidence from diverse fields to explore the possibility that NGB may represent a molecular link between gut-derived signals and neuroprotective mechanisms. We summarize current knowledge of NGB biology and discuss indirect evidence indicating that microbial metabolites, dietary phytochemicals, and hormonal mediators, including estradiol, may converge on pathways associated with NGB regulation. Based on these observations, we propose a conceptual framework in which NGB could participate in gut–brain communication while emphasizing that this hypothesis requires experimental validation. By bringing together findings that have not previously been considered within a unified context, this review highlights key knowledge gaps, opens new perspectives on the potential involvement of NGB in the GBA, and provides a foundation for future mechanistic studies in neurodegenerative and neuroinflammatory disorders. Full article