Higher Estradiol Concentrations Research Articles

2‐Oxazoline‐Based Polymer for Pharmaceutical Products Adsorption in Aqueous Media

ABSTRACTWe present the synthesis and comprehensive evaluation of a novel 2‐oxazoline‐based polymer (polyPhOx) functionalized with phenyl groups, designed for the efficient adsorption of pharmaceutical contaminants from aqueous solutions. Employing living polymerization techniques (CROP) with two distinct initiators, we optimize reaction parameters including time, temperature, and initiator concentration. The rigorous characterization of the obtained polymer is depicted as well as the demonstration of its significant binding capacity for various pharmaceuticals, exploiting π–π and electrostatic interactions to achieve superior adsorption efficiency. Adsorption experiments are performed using high concentrations of model pharmaceuticals, including aspirin, ibuprofen, metoclopramide, pyramidone, oestradiol, and indomethacin, in hydroalcoholic solutions. Initial investigations confirm polyPhOx's exceptional efficacy, particularly with hydrophobic compounds such as oestradiol, achieving adsorption capacities of up to 56.67 mg g−1. Further validation in real environmental samples highlights the polymer's practical applicability, showing substantial removal rates even under complex matrix conditions and bringing hope for effective water purification solutions. Remarkably, polyPhOx retains high adsorption performance after 10 cycles, underscoring its potential for sustainable and cost‐effective water purification. This study not only introduces a promising material for environmental remediation but also demonstrates its robustness and reusability, paving the way for advanced wastewater treatment solutions targeting persistent pharmaceutical pollutants.

High estrogen induces trans-differentiation of vascular smooth muscle cells to a macrophage-like phenotype resulting in aortic inflammation via inhibiting VHL/HIF1a/KLF4 axis.

Estrogen is thought to have a role in slowing down aging and protecting cardiovascular and cognitive function. However, high doses of estrogen are still positively associated with autoimmune diseases and tumors with systemic inflammation. First, we administered exogenous estrogen to female mice for three consecutive months and found that the aorta of mice on estrogen develops inflammatory manifestations similar to Takayasu arteritis (TAK). Then, in vitro estrogen intervention was performed on mouse aortic vascular smooth muscle cells (MOVAS cells). Stimulated by high concentrations of estradiol, MOVAS cells showed decreased expression of contractile phenotypic markers and increased expression of macrophage-like phenotypic markers. This shift was blocked by tamoxifen and Krüppel-like factor 4 (KLF4) inhibitors and enhanced by Von Hippel-Lindau (VHL)/hypoxia-inducible factor-1α (HIF-1α) interaction inhibitors. It suggests that estrogen-targeted regulation of the VHL/HIF-1α/KLF4 axis induces phenotypic transformation of vascular smooth muscle cells (VSMC). In addition, estrogen-regulated phenotypic conversion of VSMC to macrophages is a key mechanism of estrogen-induced vascular inflammation, which justifies the risk of clinical use of estrogen replacement therapy.

Biochemical profiling of the follicular environment to predict oocyte competence in cattle.

To identify markers of oocyte competence, we compared the biochemical characteristics of fluid and cells from follicles containing oocytes with different capacities to form an embryo. Follicles (5-6 mm) were dissected, and follicular fluid (FF), granulosa cells (GC), cumulus cells (CC) from immature and mature cumulus-oocyte-complexes (COC) were individually collected. The oocytes were matured, fertilized, and cultured individually until day 8 (D8) of development. On D8, the samples were grouped according to embryo production into those that gave rise to blastocysts (EMB) and those that did not reach the blastocyst stage (NEMB). In CCs from immature and mature COCs and GCs, expression of CASP3, SERPINE2, VCAN, LUM, FSHR, EGFR, PGR, and GHR genes was quantified. Cell-free DNA (cfDNA), progesterone, and estradiol concentrations in the FF were determined. Data were analyzed by Mann-Whitney U test (GraphPad Prism 9). GHR was highly expressed in immature CCs from the EMB group, whereas CASP3 was highly expressed in mature CCs from the NEMB group (P<0.05). During maturation, the expression of CASP3 and GHR genes increased only in the NEMB group. ART2 cfDNA was highly detected in FF of the NEMB compared to the EMB group. Progesterone concentration was similar between the groups, whereas estradiol concentration was higher (P<0.05) in the EMB than in the NEMB group. It was concluded that a higher level of GHR transcripts in immature CCs, lower CASP3 expression in CCs from matured COCs, lower levels of ART2, and higher estradiol concentrations in FF may indicate oocytes with greater potential for development.

Effect of baseline oestradiol serum concentration on the efficacy of anastrozole for preventing breast cancer in postmenopausal women at high risk: a case-control study of the IBIS-II prevention trial

An increased risk of breast cancer is associated with high serum concentrations of oestradiol and testosterone in postmenopausal women, but little is known about how these hormones affect response to endocrine therapy for breast cancer prevention or treatment. We aimed to assess the effects of serum oestradiol and testosterone concentrations on the efficacy of the aromatase inhibitor anastrozole for the prevention of breast cancer in postmenopausal women at high risk. In this case-control study we used data from the IBIS-II prevention trial, a randomised, controlled, double-blind trial in postmenopausal women aged 40-70 years at high risk of breast cancer, conducted in 153 breast cancer treatment centres across 18 countries. In the trial, women were randomly assigned (1:1) to receive anastrozole (1 mg/day, orally) or placebo daily for 5 years. In this pre-planned case-control study, the primary analysis was the effect of the baseline oestradiol to sex hormone binding globulin (SHBG) ratio (oestradiol-SHBG ratio) on the development of all breast cancers, including ductal carcinoma in situ (the primary endpoint in the trial). Cases were participants in whom breast cancer was reported after trial entry and until the cutoff on Oct 22, 2019, and who had valid blood samples and no use of hormone replacement therapy within 3 months of trial entry or during the trial. For each case, two controls without breast cancer were selected at random, matched on treatment group, age (within 2 years), and follow-up time (at least that of the matching case). For each treatment group, we applied a multinominal logistic regression likelihood-ratio trend test to assess what change in the proportion of cases was associated with a one-quartile change in hormone ratio. Controls were used only to determine quartile cutoffs. Profile likelihood 95% CIs were used to indicate the precision of estimates. A secondary analysis also investigated the effect of the baseline testosterone-SHBG ratio on breast cancer development. We also assessed relative benefit of anastrozole versus placebo (calculated as 1 - the ratio of breast cancer cases in the anastrozole group to cases in the placebo group). The trial was registered with ISRCTN (number ISRCTN31488319) and completed recruitment on Jan 31, 2012, but long-term follow-up is ongoing. 3864 women were recruited into the trial between Feb 2, 2003, and Jan 31, 2012, and randomly assigned to receive anastrozole (n=1920) or placebo (n=1944). Median follow-up time was 131 months (IQR 106-156), during which 85 (4·4%) cases of breast cancer in the anastrozole group and 165 (8·5%) in the placebo group were identified. No data on gender, race, or ethnicity were collected. After exclusions, the case-control study included 212 participants from the anastrozole group (72 cases, 140 controls) and 416 from the placebo group (142 cases, 274 controls). A trend of increasing breast cancer risk with increasing oestradiol-SHBG ratio was found in the placebo group (trend per quartile 1·25 [95% CI 1·08 to 1·45], p=0·0033), but not in the anastrozole group (1·06 [0·86 to 1·30], p=0·60). A weaker effect was seen for the testosterone-SHBG ratio in the placebo group (trend 1·21 [1·05 to 1·41], p=0·011), but again not in the anastrozole group (trend 1·18 [0·96 to 1·46], p=0·11). A relative benefit of anastrozole was seen in quartile 2 (0·55 [95% CI 0·13 to 0·78]), quartile 3 (0·54 [0·22 to 0·74], and quartile 4 (0·56 [0·23 to 0·76]) of oestradiol-SHBG ratio, but not in quartile 1 (0·18 [-0·60 to 0·59]). These results suggest that serum hormones should be measured more routinely and integrated into risk management decisions. Measuring serum hormone concentrations is inexpensive and might help clinicians differentiate which women will benefit most from an aromatase inhibitor. Cancer Research UK, National Health and Medical Research Council (Australia), Breast Cancer Research Foundation, and DaCosta Fund.

Sphingosine-1-phosphate mediates FSH-induced cell viability but not steroidogenesis in bovine granulosa cells

Follicle-stimulating hormone (FSH) stimulates the proliferation, survival, and estradiol synthesis of granulosa cells by binding to their G protein-coupled receptors. Although FSH activates sphingosine kinase-1 (SPHK1) to induce sphingosine-1-phosphate (S1P) synthesis, which is required to mediate the proliferative and survival effect of this gonadotrophin, the mechanisms, and the role of S1P in estradiol synthesis have not been reported. This study aimed to evaluate the importance of FSH-induced S1P synthesis as a mediator of the effects of this gonadotrophin on granulosa cell viability and steroidogenesis and to determine if FSH-induced S1P synthesis depends on estradiol, cAMP, PKA, or PKC. To achieve these objectives, we tested the effects of FSH, a sphingosine kinase-1 inhibitor (SKI-178), estradiol and inhibitors of aromatase, cAMP, PKA, and PKC (Formestane, MDL-12330A, H-89 dihydrochloride hydrate and Calphostin C respectively), on granulosa cell viability, S1P and estradiol production, and the mRNA expression of CYP19A1 and STAR in four in vitro culture experiments. The addition of FSH (1 ng/mL) increased (P < 0.05) granulosa cells number and S1P concentration in the culture media. Conversely, the addition of SKI-178 (10 μM) reduced (P < 0.05) S1P concentration negating the effect of FSH on cell viability. Inhibition of PKC and PKA, but not cAMP, reduced (P < 0.05) S1P secretion of FSH treated granulosa cells. It is important to note that the reduction in S1P secretion was strong (49 %) with the use of the PKC inhibitor. The use of formestane (10 μg) did not modify (P > 0.05) S1P secretion in FSH-treated cells; however, the addition of 5 or 10 ng/mL of estradiol increased (P < 0.05) S1P secretion. Finally, FSH increased (P < 0.05) estradiol concentration in the culture media, but this effect was not blocked by the inhibition of S1P synthesis. Similarly, FSH, SKI-178 or their combination did not modify the mRNA expression of CYP19A1 and STAR. In conclusion, S1P synthesis is stimulated FSH in granulosa cells and mediated mainly by PKC. S1P in turn promotes the granulosa cell viability, however, this does not influence estradiol synthesis. Additionally, estradiol synthesis induced by FSH is not essential for S1P synthesis, however high estradiol concentration may stimulate S1P production by granulosa cells.

Quantification of host proteomic responses to genotype 4 hepatitis E virus replication facilitated by pregnancy serum

BackgroundHepatitis E virus (HEV) infection is a common cause of acute hepatitis worldwide and causes approximately 30% case fatality rate among pregnant women. Pregnancy serum (PS), which contains a high concentration of estradiol, facilitates HEV replication in vitro through the suppression of the PI3K–AKT–mTOR and cAMPK–PKA–CREB signaling pathways. However, the proteomics of the complex host responses to HEV infection, especially how PS facilitates viral replication, remains unclear.MethodsIn this study, the differences in the proteomics of HEV-infected HepG2 cells supplemented with fetal bovine serum (FBS) from those of HEV-infected HepG2 cells supplemented with serum from women in their third trimester of pregnancy were quantified by using isobaric tags for relative and absolute quantification technology.ResultsA total of 1511 proteins were identified, among which 548 were defined as differentially expressed proteins (DEPs). HEV-infected cells supplemented with PS exhibited the most significant changes at the protein level. A total of 328 DEPs, including 66 up-regulated and 262 down-regulated proteins, were identified in HEV-infected cells supplemented with FBS, whereas 264 DEPs, including 201 up-regulated and 63 down-regulated proteins, were found in HEV-infected cells supplemented with PS. Subsequently, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that in HEV-infected cells, PS supplementation adjusted more host genes and signaling pathways than FBS supplementation. The DEPs involved in virus–host interaction participated in complex interactions, especially a large number of immune-related protein emerged in HEV-infected cells supplemented with PS. Three significant or interesting proteins, including filamin-A, thioredoxin, and cytochrome c, in HEV-infected cells were functionally verified.ConclusionsThe results of this study provide new and comprehensive insight for exploring virus–host interactions and will benefit future studies on the pathogenesis of HEV in pregnant women.

Multi-omics analysis reveals gut microbiota-ovary axis contributed to the follicular development difference between Meishan and Landrace × Yorkshire sows

BackgroundThe mechanism by which Meishan (MS) sows are superior to white crossbred sows in ovarian follicle development remains unclear. Given gut microbiota could regulate female ovarian function and reproductive capacity, this study aimed to determine the role of gut microbiota-ovary axis on follicular development in sows.MethodsWe compared the ovarian follicular development, gut microbiota, plasma metabolome, and follicular fluid metabolome between MS and Landrace × Yorkshire (L × Y) sows. A H2O2-induced cell apoptosis model was used to evaluate the effects of multi-omics identified metabolites on the apoptosis of porcine ovarian granulosa cells in vitro.ResultsCompared with L × Y sows, MS sows have greater ovary weight and improved follicular development, including the greater counts of large follicles of diameter ≥ 5 mm, secondary follicles, and antral follicles, but lesser atretic follicles. The ovarian granulosa cells in MS sows had alleviated apoptosis, which was indicated by the increased BCL-2, decreased caspases-3, and decreased cleaved caspases-3 than in L × Y sows. The ovarian follicular fluid of MS sows had higher concentrations of estradiol, progesterone, follicle-stimulating hormone, luteinizing hormone, and insulin like growth factor 1 than L × Y sows. Gut microbiota of MS sows formed a distinct cluster and had improved alpha diversity, including increased Shannon and decreased Simpson than those of L × Y sows. Corresponding to the enhanced function of carbohydrate metabolism and elevated short-chain fatty acids (SCFAs) in feces, the differential metabolites in plasma between MS and L × Y sows are also mainly enriched in pathways of fatty acid metabolism. There were significant correlations among SCFAs with follicular development, ovarian granulosa cells apoptosis, and follicular fluid hormones, respectively. Noteworthily, compared with L × Y sows, MS sows had higher follicular fluid SCFAs concentrations which could ameliorate H2O2-induced porcine granulosa cells apoptosis in vitro.ConclusionMS sows have more secondary and antral follicles, but fewer atretic follicles and apoptotic ovarian granulosa cells, as well as harbored a distinctive gut microbiota than L × Y sows. Gut microbiota may participate in regulating ovarian follicular development via SCFAs affecting granulosa cells apoptosis in sows.

Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocytes.

The present study examined the effect of polysaccharides gels made of xanthan gum and locust bean gum (gel culture system) on oocyte maturation and explored the molecules causing the beneficial effect of the gel culture system. Oocytes and cumulus cells complexes were collected from slaughterhouse-derived ovaries and cultured on a plastic plate or gel. The gel culture system improved the rate of development to the blastocyst stage. The oocytes that matured on the gel contained high lipid contents and F-actin formation, and the resultant 8-cell stage embryos had low DNA methylation levels compared to their plate counterparts. RNA sequencing of the oocytes and embryos revealed the differentially expressed genes between the gel and plate culture systems, and upstream regulator analysis revealed estradiol and TGFB1 as top activated upstream molecules. The medium of the gel culture system contained higher concentrations of estradiol and TGFB1 than that of the plate cultures system. Supplementation of the maturation medium with either estradiol or TGFB1 resulted in high lipid content in oocytes. In addition, TGFB1 improved the developmental ability of the oocytes and increased F-actin content while reducing DNA methylation levels in the 8-cell stage embryos. In conclusion, the gel culture system is useful for embryo production, potentially through the upregulation of TGFB1.

Blood hormone concentrations as markers and predictors of porcine meat production and quality?

Context The applicability of hormones in the prediction of meat production in non-ruminant species has been poorly examined. Aims To examine the applicability of plasma concentrations of leptin, estradiol, progesterone and insulin-like growth-factor I (IGF-I) for characterisation and early prediction of the production and quality of meat in gilts. Methods Animals were compared at 150 days of age with below (‘low’) and above (‘high’) median hormone concentrations and their traits of meat production and quality at the age of 200 days. Key results Animals with a high leptin concentration in blood at the age of 150 days had higher slaughter and carcass weights and weight of ham, but a lower lean meat content at the age of 200 days, than did the gilts with a low leptin concentration. Animals with a high estradiol concentration had a higher carcass weight, weight of ham and backfat thickness and a lower electrical conductivity of musculus semimembranosus than did animals with a low estradiol concentration. High blood IGF-I concentration was associated with a higher slaughter weight, carcass weight and lean meat content. No significant differences between the animals with ‘high’ and ‘low’ progesterone concentration and meat characteristics have been found. Conclusions These observations confirm the involvement of leptin, IGF-I and estradiol in up-regulation of meat production in pigs. Furthermore, they suggest that leptin can be useful for characterisation and prediction of fat production, whereas IGF-I and estradiol (but not progesterone) could be both markers and predictors of porcine growth and meat production. Implications The blood concentrations of these hormones could help in characterisation and selection of pigs for meat production.

Treatment safety of ART cycles with extremely high oestradiol concentrations using GnRH agonist trigger

Research questionAre IVF treatments with extremely high peak oestradiol levels and gonadotrophin releasing hormone (GnRH) agonist trigger associated with higher complication rates? DesignA retrospective cohort study including patients from two large medical centres treated between 2019 and 2021. A study group with extremely high peak oestradiol levels (≥20,000 pmol/l on the day of ovarian stimulation, or ≥15,000 pmol/l on the previous day) and a control group with normal range oestradiol levels (3000–12000 pmol/l) that received GnRH agonist triggering. Patients were surveyed about complaints and medical care related to ovum retrieval and medical files were reviewed. Major complication rates and the need for medical assistance were compared. ResultsSeveral differences between the study and control group were observed because of the study design: mean age was 33.01 ± 5.14 versus 34.57 ± 4.52 (P < 0.001), mean peak oestradiol levels was 26645.34 ± 8592.57 pmol/l versus 7229.75 ± 2329.20 pmol/l (P < 0.001), and mean number of oocytes were 27.55 ± 13.46 versus 11.67 ± 5.76 (P < 0.001) for the study and control group, respectively. Major complications and hospitalization rates were similar between the study and control groups (three [1.25%] versus one [0.48%]; P = 0.62 and three [1.25%] versus two [0.96%]; P = 1.0, respectively). Thirty-six patients (15.1%) in the study group and 11 (5.3%) in the control group sought medical care after retrieval, mostly due to abdominal pain, without the need for further workup or hospitalization (P < 0.001). ConclusionsExtremely high oestradiol levels were not associated with thromboembolic events, higher major complication or hospitalization rates, and therefore may be considered safe. Nevertheless, patients may be informed of possible higher rates of discomfort, mostly abdominal pain. Larger studies are warranted to confirm our results.

Neural activation associated with maternal and aversive interactions with pups in the Mongolian gerbil (Meriones unguiculatus)

According to approach-avoidance model, virgin female laboratory rats display maternal behaviour when the tendency to approach and interact with the pup is stronger than avoiding it. A positive neural mechanism that includes the medial preoptic area (mPOA)/bed nucleus of the stria terminalis (BNST) and a negative mechanism that involves the anterior hypothalamic nucleus (AHN)/ventromedial nucleus (VMN)/ periaqueductal grey (PAG) underlie to these behaviours. Unlike virgin rats, which avoid the pups, virgin females Mongolian gerbils (Meriones unguiculatus) can be immediately either maternal or aggressive with the pups. Furthermore, the Mongolian gerbil is monogamous and biparental species. Despite these difference, we hypothesised that maternal and aggressive interaction with the pups could activate mPOA/BNST and AHN/VMH/PAG, respectively, and that maternal response could be associated with high concentrations of estradiol (E2). Twenty virgin maternal females and 20 aggressive toward the pups were selected. Ten maternal females interacted with the pups (MAT-pups) and 10 with candy (MAT-candy). Of the 20 aggressive females, 10 interacted with the pups (AGG-pups) and 10 with candy (AGG-candy). Immediately after the test, blood samples were taken to quantify E2. The brains were dissected for c-Fos immunohistochemistry. MAT-pups females had significantly higher activation in mPOA/BNST than MAT-candy females, while AGG-pups showed significant activation in AHN/VMH/PAG compared with AGG-candy females. The maternal response was associated with high concentrations of E2. These results suggested a positive and a negative mechanism in the regulation of maternal behaviour in the Mongolian gerbil, and that the immediate maternal response could be due to high E2 concentrations.

High concentration of estradiol has a negative correlation with free thyroxine during the second trimester of pregnancy.

ObjectiveTo explore the relationship between estradiol (E2) and thyroid function during the second trimester of pregnancy and the effect of E2 on sodium iodide transporter (NIS) expression in cultured thyroid cells.Materials and methodsWe analyzed relationships between E2 and thyroid function in 196 pregnant women during the second trimester. Multiple linear regression analysis was performed between E2 and thyroid function. The human thyroid Nthy-ori3-1 cells were cultured in different E2 concentrations, and the mRNA levels of NIS, estrogen receptor (ER)-α, and ER-β were measured by quantitative real-time PCR. Their protein levels were assessed by western blot.ResultsE2 was positively correlated with thyroid-stimulating hormone (TSH) and negatively correlated with free thyroxine (FT4) (P < 0.05). When we corrected for age, BMI, alanine aminotransferase, and serum creatinine, E2 was still negatively correlated with FT4 (P < 0.5) during the second trimester. In Nthy-ori3-1 cells treated with 10 nM E2, NIS and ER-β mRNA levels were significantly reduced, while ER-α mRNA level was not altered (P > 0.5). Moreover, 10 nM E2 significantly decreased protein levels of ER-β, phosphorylated versions of protein kinase A (p-PKA), phosphorylated versions of cAMP response element-binding protein (p-CREB), and NIS, while treatment with the ER-β inhibitor restored the expression of p-PKA, p-CREB, and NIS (P < 0.05).ConclusionHigh concentration of E2 has a negative correlation with FT4. High concentration of E2 can inhibit the NIS expression through the ER-β-mediated pathway, which may cause thyroid hormone fluctuations during pregnancy.

Physiological interindividual variability in endogenous estradiol concentration does not influence adipose tissue and hepatic lipid kinetics in women.

Increased triglyceride (TG) and apolipoprotein B-100 (apoB-100) concentrations in plasma are important risk factors for cardiovascular disease in women. Administration of some estrogen preparations raises plasma TG and apoB-100 concentrations by increasing hepatic very low-density lipoprotein (VLDL) TG and apoB-100 secretion rates. However, the influence of physiological variation in endogenous estradiol on VLDL-TG and VLDL-apoB-100 metabolism and on free fatty acid (FFA) release into plasma (the major source of fatty acids for VLDL-TG production) is not known. We measured basal VLDL-TG, VLDL-apoB-100, and plasma FFA kinetics by using stable isotopically labeled tracers in 36 eumenorrheic, premenopausal women (age: 33 ± 2 years, BMI: 31 ± 1 kg/m2; mean ± s.e.m.) during the follicular phase of the menstrual cycle; participants were divided into two groups based on low (n = 18) or high (n = 18) plasma estradiol concentrations (defined as below or above the median value of 140 pmol/L in the whole group). Mean plasma estradiol concentration was >3-fold higher in the high-estradiol than in the low-estradiol group (299 ± 37 and 96 ± 7 pmol/L, P < 0.001); there was no difference in plasma progesterone concentrations between the two groups (P = 0.976). There were no significant differences in plasma FFA concentration, FFA rate of appearance in plasma, VLDL-TG and VLDL-apoB-100 concentrations, hepatic VLDL-TG and VLDL-apoB-100 secretion rates, VLDL-TG and VLDL-apoB-100 plasma clearance rates, and mean residence times (all P ≥ 0.45). No significant associations were found between plasma estradiol concentration and FFA, VLDL-TG, and VLDL-apoB-100 concentrations and kinetics (all P > 0.19). Plasma estradiol concentration is not an important correlate of basal plasma FFA, VLDL-TG, and VLDL-apoB-100 kinetics in premenopausal women.

High steroid content in conditioned medium of granulosa cells may disrupt primordial follicles formation in in vitro cultured one-day-old murine ovaries

This study was conducted to investigate the main and interactive effects of two methods of culture medium preparation [base medium vs granulosa cells conditioned medium (GCCM)] and two nutrient supplements [fetal bovine serum (FBS) vs knock-out serum replacement (KSR)] on formation and activation of primordial follicles and gene expression of corresponding factors during a seven-day culture period. One-day-old mouse ovaries were cultured with four different culture media including base medium containing FBS (BMF), base medium containing KSR (BMK), GCCM prepared with FBS (CMF) and GCCM prepared with KSR (CMK), and samples for histological and molecular assessments were collected on days 3 and 7 of culture. Further, steroid content of media was measured. Histological examination showed that KSR enhanced follicular formation and the number of follicular count was greater in BMK than CMF group (P < 0.05). Moreover, follicular activation was higher in CMK group than BMK and CMF groups (P < 0.05). Additionally, RT-PCR revealed that KSR upregulated Gdf9 gene expression (P < 0.05), while GCCM diminished expression of Gdf9, Bmp15, Notch2, Figla and Foxl2 (P < 0.05). GCCM decreased expression of Pten and increased expression of Pi3k (P < 0.05). Besides, hormonal assays indicated higher concentrations of estradiol and progesterone in GCCM compared with base media (P < 0.0001). In conclusion, the present study showed base medium containing KSR could serve as a proper medium for in vitro culture of neonatal mouse ovary since it could better support formation of primordial follicles. Yet BMK did not promote follicular activation as well as GCCM prepared with KSR did, and therefore, requires modifications.

The relationship between milk oestradiol concentrations and oestrus activity in lactating Holstein–Friesian cows

Context Detecting oestrus to facilitate the optimal timing of artificial insemination is key to optimal reproductive performance in dairy cows. Aims The aims of the present study were to investigate the relationship between activity and milk oestradiol concentrations during oestrus. Methods Accelerometers (IceQubes, IceRobotics Ltd, UK) were used to monitor the activity (the number of steps per day) of 37 lactating Holstein–Friesian cows during the peri-oestrus period. Daily milk samples were analysed for oestradiol and progesterone concentrations. Key results An increase in activity sufficient to generate an oestrus alert (behavioural oestrus) from an activity sensor (GEA Rescounter II) was detected in 29 of 37 (78%) cows. Milk oestradiol increased from 2.0 ± 0.5 pg/mL 4 days before oestrus to 8.3 ± 1.1 pg/mL on the day of behavioural oestrus, then fell to 2.6 ± 0.6 pg/mL by 3 days afterwards. Similarly, activity also increased and peaked on the day of oestrus. However, in 8 of 37 (22%) cows, although milk oestradiol concentrations increased following the fall in progesterone, there was no significant increase in activity (silent oestrus). Milk oestradiol concentrations were positively correlated (r = 0.37; P = 0.03) with activity on the day of oestrus. Cows that subsequently became pregnant took a similar number of steps (2806 ± 282.3 vs 2850 ± 372.5 steps for PD+ vs PD− cows respectively) but had higher oestradiol concentrations (11.2 ± 2.06 vs 5.1 ± 0.51 pg/mL for PD+ vs PD− cows respectively) on the day of oestrus than did cows that did not become pregnant. Conclusions Increases in oestradiol concentrations above a threshold of approximately 10 pg/mL are associated with an increased activity, and higher oestral oestradiol concentrations are associated with a higher rate of pregnancy following insemination. Implications This study has demonstrated the significance of oestradiol concentration to the fertility of dairy cows. Thus, strategies to enhance oestradiol secretion may enhance fertility. Further, milk oestradiol measures may be developed as an aid to oestrus detection in lactating dairy cows, although further studies are required to determine whether on-farm assessment of milk oestradiol concentration can be developed to add to the repertoire of biomarkers of oestrus in lactating cattle.

Recipient sex and estradiol levels affect transplant outcomes in an age-specific fashion

Sex-specific influences have been shown for a variety of diseases. Whether donor or recipient sex and sex hormone levels impact alloimmune responses remains unclear. In unifactorial and multifactorial analyses of more than 400 000 SRTR listed kidney transplant patients, we found that younger female recipients had an inferior death-censored graft survival that was independent of donor sex. In contrast, graft survival was superior in older female recipients, suggesting the impact of recipient sex hormones over chromosomal sex mismatches. Those clinical changes were delineated in experimental skin and heart transplant models showing a prolongation of graft survival in ovariectomized young female recipients. In contrast, graft survival was comparable in ovariectomized and naïve old female recipients. Young ovariectomized mice showed reduced amounts and a compromised T cell proliferation. Deprivation of female hormones dampened the production of interferon (IFN)-γ and interleukin (IL)-17+ by CD4+ T cells while augmenting systemic counts of Tregs. Increasing estradiol concentrations in vitro promoted the switch of naïve CD4+ T cells into Th1 cells; high physiological estradiol concentrations dampening Th1 responses, promoted Tregs, and prolonged graft survival. Thus, clinical observations demonstrate age-specific graft survival patterns in female recipients. Estrogen levels, in turn, impact the fate of T cell subsets, providing relevant and novel information on age- and sex-specific alloimmunity.

LOW DOSE HCG IN PREVENTING OHSS IN HIGH-RISK WOMEN

Ovarian hyperstimulation syndrome(OHSS) is a complication of fertility treatment, which uses pharmacological ovarian stimulation to increase the number of oocytes and therefore embryos available during assisted reproductive technology (ART).Severe ovarian hyperstimulation syndrome (OHSS) is well known to be a rare but potentially fatal condition in anovulatory women with polycystic ovarian syndrome (PCOS) when undergoing IVF. Low-dose stimulation is thus recommended,but it can still lead to ovarian hyperstimulation associated with high serum oestradiol concentrations by the time leading follicles reach maturity. Several methods have, therefore, been applied to prevent OHSS. First, risk assessment is made on the basis of the previous history of OHSS and the identification of women with PCO. Second, in treatment cycles a high concentration of oestradiol and three ultrasound parameters (i.e. high number of follicles, large ovarian volume, and high stromal vascularity) on the day of human chorionic gonadotrophin (HCG) are all predictive of increased risk of developing OHSS.

Metabolic and Hormonal Profile of Adolescent Girls With Polycystic Ovary Syndrome With Concomitant Autoimmune Thyroiditis.

IntroductionBoth polycystic ovary syndrome (PCOS) and autoimmune thyroiditis (AT) are considered to be among the most common endocrinopathies in young women, and they are classified as diseases that affect many processes in the human body. Their role in the development of metabolic disorders and diseases of the cardiovascular system in adult women is also emphasized. However, there are no data available to assess such risk in the teenage girl population. The aim of the study was to assess the hormonal and metabolic profile of adolescent girls with PCOS, additionally diagnosed with AT, as well as to identify possible risk factors for the coexistence of AT and PCOS.Material and Methods80 euthyroidic PCOS patients were qualified for the study (chronological age 16.54 ± 1.00 years, BMI 24.60 ± 4.16 kg/m2). Eighteen girls diagnosed with AT were included in the study group and 62 girls without AT—in the control group. Each patient had biochemical and hormonal tests performed. Additionally, to diagnose AT, the level of antibodies against thyroid peroxidase (anti-TPO) and anti-thyroglobulin (anti-TG), as well as the image of the thyroid gland on ultrasound examination, were taken into account.ResultsEstradiol concentration was significantly higher in the study than in the control group (203.00 ± 217.00 vs. 152.00 ± 78.50 pmol/L, p=0.02). Higher DHEAS concentrations were also observed in the AT group compared with the group without AT (391.28 ± 176.40 vs. 317.93 ± 114.27 µg/dl, p=0.04). Moreover, there was a positive correlation between AT and estradiol concentration (ry=0.27; p=0.04). It was also shown that there is a tendency toward statistical significance for the positive correlation between the positive anti-TPO titer and the glucose concentration at 120 min OGTT (rƴ=0.26; p=0.07) and girls with PCOS and AT had higher glucose levels in 120 min OGTT (115.29±41.70 vs. 98.56±28.02 mg/dl, p=0.08).ConclusionThe study results showed no difference in the metabolic profile between the groups. The high concentration of estradiol found in girls with PCOS and AT may indicate the role of this hormone in the development of the autoimmune process. However, the numbers are small, and more research is needed to confirm our findings.

Assessment of Estradiol-Induced Gonadotropin Surge Generation: Preliminary Results From a Protocol of Graded Transdermal Estradiol Dosing

In normally-cycling women, high midcycle estradiol (E2) concentrations (> 200-300 pg/ml) derived from the dominant follicle trigger an approximately 10-fold increase in LH and 4-fold increase in FSH levels—the gonadotropin surge—over about 48 hours. In prenatally-androgenized female rats and sheep (animal models with polycystic ovary syndrome [PCOS]-like features), high-dose estrogen fails to initiate gonadotropin surges. However, it remains unclear whether E2-induced gonadotropin surge generation is defective in PCOS. Baird et al., induced LH surges in 12 women with PCOS and 6 normally-cycling controls (studied in the early follicular phase) with 3 days of high-dose oral ethinyl E2, and the LH peak magnitude was similar in both groups (JCEM 1977;45:798-801). However, the dose of ethinyl E2 in this study (200 mcg/day) was markedly supraphysiological, and the dosing schedule did not mimic the normal, gradual increase in E2 levels observed in normal follicular phases. To further assess potential impairments in E2 augmentation of gonadotropin secretion in PCOS, we developed a study protocol involving graded transdermal E2 dosing (with dose adjustments as needed) to gradually achieve serum E2 concentrations approximating 300-400 pg/ml. Herein we present our early experience with this protocol in six normally-cycling, non-hyperandrogenic women: median (range) age 23.1 (19.8–29.0) years, body mass index 20.7 (19.2–27.2) kg/m2, calculated serum free testosterone 2.7 (2.4–5.3) pg/ml. Study subjects began transdermal E2 on cycle day 4 with the following dose-escalation protocol: 0.2 mg/d x 1 day, then 0.3 mg/d x 1 day, then 0.4 mg/d x 1 day, then 0.5 mg/d. Daily serum E2 measurements informed dose adjustments (e.g., a measured E2 level > 400 pg/ml prompted a dose reduction; a measured E2 level < 250 pg/ml while on the 0.5 mg/d dose prompted a dose increase to 0.6 mg/d). In these six women, daily E2 values (pg/ml) were as follows: baseline 31 (17–36) (median [IQR]) on cycle day 4 (the morning of E2 initiation); 205 (135–240) on day 5; 159 (135–270) on day 6; 263 (202–363) on day 7; 276 (239–351) on day 8; 328 (307–367) on day 9; 333 (269–402) on day 10; and 328 (253–354) on day 11. Median transdermal E2 doses (mg/d) on days 5 through 11 were 0.2, 0.3, 0.4, 0.35, 0.4, 0.4, and 0.4, respectively. Morning serum LH concentrations (median [IQR]) were 5.2 (3.5–6.1) IU/L immediately before E2 initiation (cycle day 4), decreased to a nadir of 1.5 (1.0–2.4) IU/L after 2 days of E2 (cycle day 6), thereafter increasing 8.1-fold to a peak of 12.1 (8.1–18.3) IU/L after 5 days of E2 (cycle day 9). FSH similarly changed from baseline median 3.5 (3.4–4.2) IU/L to nadir median of 1.6 (1.2–1.8) IU/L after 2 days of E2, thereafter increasing again to 3.5 (2.6–4.0) IU/L after 5 days of E2. We conclude that this experimental protocol may be useful to investigate potential defects in E2-induced LH surge generation in PCOS.

Non-Peutz–Jeghers syndrome-associated ovarian sex cord tumor with annular tubules treated by radiotherapy: a case report and literature review

There are no standard treatment options for metastatic and recurrent non-Peutz–Jeghers syndrome (PJS)-associated sex cord tumor with annular tubules (SCTAT). The effects of chemotherapy and/or radiotherapy are still not well-defined. Herein, we present a case of a metastatic and recurrent non-PJS-associated SCTAT showing high serum estradiol and progesterone concentrations after surgery and chemotherapy. Radiotherapy (50 Gy/25 fractions) triggered a sharp reduction in the sizes of the metastatic and recurrent masses, and estradiol and progesterone concentrations. Accordingly, we consider that radiotherapy might be effective and safe for metastatic and recurrent SCTAT. The roles of radiotherapy in non-PJS SCTAT should be further validated in large-scale prospective clinical trials.