Estradiol-17β Concentrations Research Articles

Differential effects of chronic 17β-oestradiol treatment on rat behaviours relevant to depression.

Sex differences are a prominent feature of the pathophysiology of psychiatric disorders, such as major depressive disorder, which affects women at a higher incidence than men. Research suggests that the most potent endogenous oestrogen, 17β-oestradiol, may have therapeutic potential in treating depression. However, preclinical studies have produced mixed results, likely as a result of various methodological factors such as treatment duration. The present study aimed to investigate the effects of ovariectomy and chronic 17β-oestradiol treatment via a s.c. silastic implant on behaviours relevant to depression in adult female Sprague-Dawley rats. Rats were assessed in the forced swim test, saccharin preference test and novel object recognition memory test, as well as for possible confounding behaviours, including locomotion and anxiety (open field test) and motivation and anxiety (novelty suppressed feeding test). Treatment effects were verified using body and uterus weight, as well as serum concentrations of 17β-oestradiol, progesterone and testosterone. Compared to ovariectomised rats, chronic 17β-oestradiol treatment enhanced saccharin preference and novel object recognition performance. There were no group differences in passive or active coping behaviour when assayed using the forced swim test. Taken together, these results support an antidepressant-like action of oestrogens but highlight that the beneficial effects of chronic 17β-oestradiol treatment may be related to specific depression-related symptoms, particularly anhedonia and memory.

Maternal estradiol and progesterone concentrations among singleton spontaneous pregnancies during the first trimester.

Pregnancy hormonal milieu represents a crucial determinant of fetal outcome. We aim to determine 17β-estradiol (E2) and progesterone (PGT) concentrations in spontaneous pregnancies during the first trimester. In addition, we aim to determine E2 concentrations as a function of gestational age (GA) and PGT. Between November 2015 and March 2017, 104 healthy women of at least 18years undergoing medical consultation for voluntary interruption of pregnancy were enrolled in an observational study at University Hospital ASST Fatebenefratelli Sacco, Milan, Italy. Only singleton pregnancies between 5+0 and 13+6 weeks of gestation were eligible. First trimester ultrasound scans were performed for dating and one fasting venous blood sample was collected for E2 and PGT determinations. E2 and PGT concentrations steadily increased according to GA. The correlation between E2 on a logarithmic scale and PGT concentrations was expressed by the following equation, explaining 12.6% of E2 variance: logE2 = 2.57 + 0.1 × PGT, (r = 0.34, p < 0.001). By performing a multivariable linear regression analysis adjusted for variables significantly correlated with E2 concentrations, we obtained a model explaining the 53.5% of E2 variance. The final equation to determine E2 concentrations among Caucasian women was: logE2 = 1.96 + 0.01 × GA + 0.004 × PGT. Gestational week-specific reference intervals are reported for maternal E2 and PGT concentrations during early pregnancy, further providing a model for E2 assessment in this period. This will represent a starting point for further evaluations between twin and ART pregnancies, as well as to potentially improve pregnancy outcome and future health of the offspring.

Joint effects of chronic exposure to environmentally relevant levels of nonylphenol and cadmium on the reproductive functions in male rockfish Sebastiscus marmoratus

Nonylphenol (NP) and Cadmium (Cd) are two common contaminants that can be detected in aquatic environments. Nevertheless, the combined toxicity of NP and Cd at environmentally relevant concentrations in aquatic organisms has not been thoroughly characterized to date. In the present study, the interactions between NP and Cd on male Sebastiscus marmoratus were studied. After 21 days of exposure, the brain aromatase activity was observed to be significantly induced by 100 ng/L NP and 40 μg/L Cd, whereas all of the concentrations of co-treatment resulted in an increase in brain aromatase activity. Additionally, NP could also reduce plasma testosterone concentration, while NP, Cd and their mixture could induce plasma 17β-estradiol (E2) concentration and VTG concentration. The interactions between NP and Cd on the reproductive physiology were antagonism. Our results also support the notion of using these indicators as biomarkers for exposure to EDCs and further extend the boundary of biomonitoring to environmental levels.

Effect of different dosages of PG-600 on ovulation and pregnancy rates in ewes during the breeding season.

This study compared the reproductive effects of different dosages of PG-600 (Intervet/Merck Animal Health, Madison, NJ) during the breeding season of ewes. PG-600 is a single-dose injectable product labeled for estrous induction in swine, containing equine chorionic gonadotropin (80 IU/mL) and human chorionic gonadotropin (40 IU/mL). PG-600 is routinely used off-label for out-of-season estrous induction in sheep. However, at the most common dose administered to ewes (5 mL), PG-600 is likely to overstimulate the ovaries, resulting in reduced pregnancy rates. Following estrous synchronization with intravaginal progesterone and cloprostenol, Polypay ewes were treated with 5 mL PG-600 (T1; n = 8), 1.5 mL PG-600 (T2; n = 8), or 5 mL saline (C; n = 8) and then mated to rams. Jugular vein samples were collected prior to the PG-600 injection (0 hr) and at 2, 4, 8, 12, and 24 hr after injection. Serum estradiol-17β was determined by chemiluminescence and among groups using repeated measures analysis of covariance. Ovulation and pregnancy rates were determined by transrectal ultrasonography and compared by one-way ANOVA and chi-square, respectively. Estradiol-17β concentrations were greater in T1 compared to T2 and C (P < 0.001). Ovulation rate was greater (P < 0.001) but pregnancy rate was lower (P < 0.001) in the T1 compared to C and T2. These data confirm that a 5 mL dose of PG-600 administered to ewes during the breeding season overstimulates the ovaries, which may then reduce fertilization or embryo survival. Future research will focus on the effects of different dosages of PG-600 on pregnancy rate of ewes during the nonbreeding season.

Occurrence of antibiotics, estrogenic hormones, and UV-filters in water, sediment, and oyster tissue from the Chesapeake Bay

Globally, the occurrence of contaminants of emerging concern (CECs) in the environment has raised critical questions on ecological and human health, but few efforts have focused on the Chesapeake Bay, the largest estuary in the United States. Here, 43 antibiotics, 3 estrogenic hormones, and 5 ultraviolet-filters (UV-filters), which are active ingredients in a variety of personal care products, were measured in water, sediment, and oyster tissue from 14 sites along the Eastern Shore of the Chesapeake Bay in Maryland. Fluoroquinolone, macrolide, and sulfonamide antibiotics were detected in water samples. As both human- and animal-labeled antibiotics were found, wastewater effluent and agricultural runoff were identified as potential sources. The highest aqueous-phase concentrations were recorded for norfloxacin (94.1 ng/L), enrofloxacin (17.8 ng/L), sulfamethoxazole (14.8 ng/L), and clarithromycin (9.7 ng/L). Estrone and four UV-filters, namely 2-ethylhexyl-4-methoxycinnamate, benzophenone-3, homosalate, and octocrylene, were frequently detected in Chesapeake Bay water (93–100%), sediment (100%), and oyster tissue (79–100%). High sediment-phase concentrations of estrone (58.4 ng/g) and 17β-estradiol (11.5 ng/g) were detected at the mouth of the Manokin River. Homosalate and benzophenone-3 were present at concentrations as high as 187.9 and 113.7 ng/L in water, 74.2 and 10.8 ng/g in sediment, and 158.3 and 118.0 ng/g in oyster tissue, respectively. These results demonstrate the ubiquitous presence of CECs in the Chesapeake Bay, confirm UV-filter bioaccumulation in oysters, and suggest the need for improved CEC removal during municipal wastewater treatment and agricultural waste management within the Chesapeake Bay watershed.

Preventative effects of resveratrol and estradiol on streptozotocin-induced diabetes in ovariectomized mice and the related mechanisms.

Resveratrol, a non-flavonoid polyphenolic compound, is structurally and functionally similar to estrogen and has drawn great attention for its potentially beneficial effects on diabetes. However, it is not known whether it shares the same protective effect against diabetes as estrogen and the underlying mechanisms. The aim of the present study was to investigate the protective effects of phytoestrogen resveratrol and exogenous 17β-estradiol against streptozotocin (STZ)-induced type 1 diabetes. Female mice were ovariectomized (OVX) and chronically injected with different concentrations of resveratrol (0.1, 1 or 10 mg/kg) and 17β-estradiol (0.01, 0.1 or 1 mg/kg) subcutaneously for 4 weeks, and the levels of blood glucose, plasma insulin, plasma antioxidant capacity, the changes of pancreatic islet cells and the expressions of glucose transporter 4 (GLUT4), insulin receptor substrate 1 (IRS-1) and phosphorylation of extracellular signal-regulated kinase (p-ERK) were detected. Resveratrol and 17β-estradiol significantly inhibited the increase of the blood glucose level and the rise of plasma malondialdehyde in STZ-induced diabetic mice, improved the levels of plasma antioxidant capacity and plasma insulin, protected the pancreatic islet cells, and increased the expressions of GLUT4 and IRS-1, but decreased p-ERK expression in skeletal muscle and myocardial tissue. The results suggest that resveratrol or 17β-estradiol shows obvious protection against STZ-induced diabetes in OVX mice, the mechanisms probably involve their ameliorating antioxidant activities and islet function, promoting muscle glucose uptake and inhibiting the expression of p-ERK.

Body condition and reproductive status of chum salmon during homing migration in Otsuchi Bay, Japan, after the 2011 Great East Japan Earthquake and Tsunami

To understand how the earthquake and tsunami on March 11 2011 affected homing chum salmon Oncorhynchus keta in Otsuchi Bay, Japan, we monitored the body condition and reproductive status at the mouth of the bay in October, November, December, and January from 2012 to 2016. The average fork length in 2014 was larger than that in 2012 and 2015, due to the high proportion of 5- and 6-year-old salmon and the low proportion of 3- and 4-year-old salmon, possibly because of the decrease in the salmon fry released by the hatchery program after the disaster. Plasma estradiol-17β (E2) concentrations in females were high in October and November and declined in December and January, whereas plasma 11-ketotestosterone concentrations in males were consistently high throughout the sampling period. Plasma 17α,20β-dihydroxy-pregnen-3-one (17α,20β-P) increased in both sexes in December and/or January. These changes were consistently observed from 2012 to 2016; however, the rise in 17α,20β-P and decline in E2 levels in female salmon in December were conspicuous in 2013 and 2014, reflecting changes in age composition. These results suggest that the earthquake and tsunami temporarily affected the age composition and the physiological condition of chum salmon returning to Otsuchi Bay.

Inhibin anti-peptide antibody macromolecule: An approach to improve fecundity in Clarias batrachus

Inhibins are members of the transforming growth factor beta (TGFβ) superfamily known to regulate ovarian functions through stimulation of the hypothalamo-pituitary-gonadal axis. In the present study, we aimed to design a species-specific inhibin-α chimeric peptide (ICP) and evaluate the effect of immunoneutralization using anti-ICP antisera to enhance the reproductive performance in female Clarias batrachus. Injection of anti-ICP antisera caused a significant increase in the number of oocytes at a medium dose (200 μl) in comparison to high dose (400 μl) and control (normal rabbit serum). Histological observations confirmed the dose-dependent advancement in oocyte maturation. Furthermore, anti-ICP antisera treated groups exhibited a significant increase in the serum concentrations of follicle stimulating hormone (FSH) and 17β-estradiol (E2) hormones. The anti-ICP antisera decreased the mRNA expression levels of inhibin-α while stimulated the transcript levels of inhibin-βA, FSHβ, CYP 19a1, 3β-HSD and StAR respectively in a dose-dependent manner. Collectively, these findings indicate that anti-ICP antibody macromolecules modulate the endogenous reproductive hormonal secretion and enhance oocyte quality and quantity in female C. batrachus. This is the first report wherein antibodies against inhibins were used to promote reproductive performances and investigate the underlying molecular mechanisms in fishes.

Serum progesterone concentrations during FSH superstimulation of the first follicular wave affect embryo production in sheep

The aim of the present study was to evaluate the effect of serum progesterone concentrations during the superstimulatory treatment of the first follicular wave on fertilization rate and embryo development in sheep. A total of 71 Merino ewes received a superstimulatory FSH treatment during Wave 1 of ovarian follicular development (Day 0 Protocol), which was administrated under low progesterone concentrations typical of the early luteal phase (control group, n = 33) or under high progesterone concentrations induced by the administration of an intravaginal device from Day 0 to Day 3 containing 0.3 g progesterone (n = 38). Intrauterine insemination after FSH superstimulation was followed by uterine flushing 6 days later. Serum progesterone concentrations from Day 0 to 3 were greater in those ewes treated with progesterone (P < 0.05), while serum estradiol-17β concentrations were not affected by the treatment. Although the mean number of corpora lutea per donor was not affected by the progesterone treatment, the number of collected ova and embryos was greater in progesterone treated than untreated ewes (6.6 ± 0.7 compared with 4.6 ± 0.9, respectively; P < 0.05). Furthermore, progesterone treatment increased fertilization rate (93.3% compared with 83.3%; P < 0.05) and the proportion of Grade 1 embryos (67.7% compared with 52.7%; P < 0.05) compared with the control group. In conclusion, oocyte fertilization rate and embryo quality are improved by high progesterone concentrations during FSH superstimulation, which suggests an important role of progesterone during preovulatory follicular development.

Impact of repeated lipopolysaccharide administration on ovarian signaling during the follicular phase of the estrous cycle in post-pubertal pigs.

Increased circulating lipopolysaccharide (LPS) results from heat stress (HS) and bacterial infection, both of which are associated with reduced female fertility. Specific effects of low-level, repeated LPS exposure on the ovary are unclear, as many studies utilize a bolus model and/or high dosage paradigm. To better understand the effects of chronic LPS exposure on ovarian signaling and function, post-pubertal gilts (n = 20) were orally administered altrenogest for 14 d to synchronize the beginning of the follicular phase of the ovarian cycle. For 5 d after synchronization, gilts (163 ± 3 kg) received IV administration of LPS (0.1 µg/kg BW, n = 10) or saline (CT, n = 10) 4× daily. Blood samples were obtained on days 1, 3, and 5 of LPS treatment. Follicular fluid was aspirated from dominant follicles on day 5, and whole ovarian homogenate was used for transcript and protein abundance analysis via quantitative real-time PCR and western blotting, respectively. There were no treatment differences detected in rectal temperature on any day (P ≥ 0.5). Administering LPS increased plasma insulin (P < 0.01), LPS-binding protein (LBP; P < 0.01), and glucose (P = 0.08) on day 1, but no treatment differences thereafter were observed (P = 0.66). There were no treatment differences in follicular fluid concentration of LBP or 17β-estradiol (P = 0.42). Gilts treated with LPS had increased abundance of ovarian TLR4 protein (P = 0.01), but protein kinase B (AKT) and phosphorylated AKT (pAKT) were unchanged and no effect of LPS on components of the phosphatidylinositol 3 kinase (PI3K) pathway were observed. There was no impact of LPS on ovarian abundance of STAR or CYP19A1, nor ESR1, LDLR, CYP19A1, CYP17A1, or 3BHSD. In conclusion, repeated, low-level LPS administration alters inflammatory but not steroidogenic or PI3K signaling in follicular phase gilt ovaries.

Plasma estradiol-17β Concentrations in Mares Treated for Experimentally Induced Ascending Placentitis

Plasma estradiol-17β Concentrations in Mares Treated for Experimentally Induced Ascending Placentitis

PO-168 Loss of the MCC gene expression promotes invasiveness of colon cancer cells

IntroductionBreast cancer is the most common cancer in women in developed countries. Many reports have suggested that oestrogens play a key role in normal mammary gland development and in breast cancer progression and metastasis. About 70% of breast carcinoma expresses the oestrogen receptor alpha. The selective oestrogen receptor modulator tamoxifen, functions as an antagonist in most ER-positive breast tumours. However, 20%–30% of tumours are resistant to tamoxifen therapy. Foetal oestrogen estetrol seems to have a protective role during pregnancy and anti-estrogenic properties on breast tissue. Estetrol is an interesting compound for therapeutic applications due to its high oral bioavailability and long terminal elimination half-life, with no active oestrogen metabolites. Traditional two dimensional (2D) cell cultures lack proper environmental context leading to changes on cell function. On the contrary, three-dimensional (3D) cell culture has gained increasing interest in drug discovery due to its realistic modelling of complex environment that drives to cellular heterogeneity, nutrient and oxygen gradients of tumours. The aim of the current study was to evaluate whether estetrol have an anti-tumour activity in T47D breast cancer cells on 2D and 3D culture systems.Material and methodsT47D cells were growth in DMEM high glucose, phenol red-free media with 10% FBS. After starvation, cells were treated with increasing concentrations (1, 10, 100, 1000 nM) of Estetrol (E4), 17β-oestradiol (E2) and Tamoxifen (Tmx). All experiments were realised in DMEN with 5% of charcoal stripped FBS. Cell proliferation (trypan blue), clonogenic and wound healing micro-inserts assays were performed on 2D cell culture. T47D spheroids were performed on 96-well low attachment plate. 3D cell proliferation was measured as Feret diameter and spheroid-based migration assay was measured as radial migration index.Results and discussionsWe observed that 100 and 1000 nM of E4 significantly reduced cell proliferation, survival fraction and migration compared to the control on 2D cell model. As expected, Tmx was more effective on the reduction of cell proliferation, survival fraction and migration. On the other side, E4 and Tmx significantly reduced 3D cell proliferation and migration compared to the control. We can conclude that E4 do not stimulate cell proliferation and migration, on the contrary reduced it.ConclusionWe can conclude that E4 could have an anti-oestrogenic activity on T47D breast cancer cell line.

The anti-androgenic effect of chronic exposure to semicarbazide on male Japanese flounder (Paralichthys olivaceus) and its potential mechanisms

Semicarbazide (SMC), a new marine pollutant, has anti-estrogenic effects on female Japanese flounder (Paralichthys olivaceus). However, whether SMC also affects the reproductive endocrine system of male marine organisms is currently unclear. In this study, Japanese flounder embryos were exposed to 1, 10, and 100 μg/L SMC for 130 days. Plasma testosterone (T) and 17β-estradiol (E2) concentrations were significantly decreased in male flounders after SMC exposure. The expression of genes involved in T and E2 synthesis, including steroidogenic acute regulatory protein, cytochrome P450 11A1, 17α-hydroxylase, 17β-hydroxysteroid dehydrogenase and cytochrome P450 19A, was down-regulated in the gonads, which may explain the decrease in plasma sex hormones levels. Moreover, SMC-mediated changes in the transcription of these steroidogenic genes were associated with reduced levels of follicle-stimulating hormone beta subunit (fshβ), luteinizing hormone beta subunit (lhβ), follicle-stimulating hormone receptor (fshr) and luteinizing hormone receptor (lhr) mRNA. In addition, down-regulated transcription of fshβ and lhβ in the SMC exposure groups was affected by reduced mRNA levels of seabream gonadotropin-releasing hormone (sbgnrh), g-protein-coupled receptor 54 (gpr54) in the kisspeptin/gpr54 system, as well as the gamma-aminobutyric acid (GABA) synthesis enzyme glutamic acid decarboxylase (gad). Overall, our results showed that environmentally relevant concentrations of SMC exerted anti-androgenic effects in male flounders via impacting HPG axis, kiss/gpr54 system and GABA synthesis, providing theoretical support for investigating reproductive toxicity of environmental pollutants that interfere with the neuroendocrine system.

Sex steroids mediate discrete effects on HDL cholesterol efflux capacity and particle concentration in healthy men

Exogenous testosterone decreases serum concentrations of high-density lipoprotein cholesterol (HDL-C) in men, but whether this alters cardiovascular risk is uncertain. To investigate the effects of testosterone and estradiol on HDL particle concentration (HDL-Pima) and metrics of HDL function. We enrolled 53 healthy men, 19 to 55years of age, in a double-blinded, placebo-controlled, randomized trial. Subjects were rendered medically castrate using the GnRH receptor antagonist acyline and administered either (1) placebo gel, (2) low-dose transdermal testosterone gel (1.62%, 1.25g), (3) full replacement dose testosterone gel (1.62%, 5g) or (4) full replacement dose testosterone gel together with an aromatase inhibitor for 4weeks. At baseline and end of treatment, serum HDL total macrophage and ABCA1-specific cholesterol efflux capacity (CEC), HDL-Pima and size, and HDL protein composition were determined. Significant differences in serum HDL-C were observed with treatment across groups (P=.01 in overall repeated measures ANOVA), with increases in HDL-C seen after both complete and partial testosterone deprivation. Medical castration increased total HDL-Pima (median [interquartile range] 19.1 [1.8] nmol/L at baseline vs 21.3 [3.1] nmol/L at week 4, P=.006). However, corresponding changes in total macrophage CEC and ABCA1-specific CEC were not observed. Change in serum 17β-estradiol concentration correlated with change in total macrophage CEC (β=0.33 per 10 pg/mL change in serum 17β-estradiol, P=.03). Testosterone deprivation in healthy men leads to a dissociation between changes in serum HDL-C and HDL CEC. Changes in serum HDL-C specifically due to testosterone exposure may not reflect changes in HDL function.

PGF2α and gonadal steroid plasma levels of successful and unsuccessful spawning Piaractus mesopotamicus (Teleostei, Characiformes) females

Gonadal steroid and prostaglandin F2α (PGF2α) plasma levels were evaluated in successfully (SP) and unsuccessfully ovulated (UN) female Piaractus mesopotamicus. Forty-one females were injected with crude carp pituitary extract (0.6 and 5.4 mg kg−1 with a 24-h interval between the doses) and sampled to determine the plasma concentration of 17β-estradiol (E2), 17α-hydroxyprogesterone (17α-OHP), 17α,20β-dihydroxy-4-pregnen-3-one (DHP), PGF2α, and testosterone (T) after each injection (first—A1 and second—A2), and at the time of ovulation for SP and UN. Two clusters were obtained using multivariate analysis: 1—composed of all A1, all A2, and some UN; and 2—composed of all SP and some UN. Median values of E2 plasma levels were similar between clusters; however, plasma levels of T, 17α-OHP, DHP, and PGF2α of cluster 2 (predominantly formed by SP) were higher than those of cluster 1. Since cluster 2 contained all SP and females of this cluster presented higher levels of PGF2α, T, 17α-OHP, and DHP, here we evidently shown in an unprecedented manner that concomitant increased levels of these substances were associated with successful ovulation in this species, but such an increase was not determinant for successful ovulation due to the presence of some UN females in the same cluster 2. These findings highlight the unexplored potential of PGF2α to be used as an accessory tool for inducing successful ovulation for fish farming purposes.

Impact of heat stress during the follicular phase on porcine ovarian steroidogenic and phosphatidylinositol-3 signaling.

Environmental conditions that impede heat dissipation and increase body temperature cause heat stress (HS). The study objective was to evaluate impacts of HS on the follicular phase of the estrous cycle. Postpubertal gilts (126.0 ± 21.6 kg) were orally administered altrenogest to synchronize estrus, and subjected to either 5 d of thermal-neutral (TN; 20.3 ± 0.5 °C; n = 6) or cyclical HS (25.4 - 31.9 °C; n = 6) conditions during the follicular phase preceding behavioral estrus. On d 5, blood samples were obtained, gilts were euthanized, and ovaries collected. Fluid from dominant follicles was aspirated and ovarian protein homogenates prepared for protein abundance analysis. HS decreased feed intake (22%; P = 0.03) and while plasma insulin levels did not differ, the insulin:feed intake ratio was increased 3-fold by HS (P = 0.02). Insulin receptor protein abundance was increased (29%; P < 0.01), but insulin receptor substrate 1, total and phosphorylated protein kinase B, superoxide dismutase 1, and acyloxyacyl hydrolase protein abundance were unaffected by HS (P > 0.05). Plasma and follicular fluid 17β-estradiol, progesterone, and lipopolysaccharide-binding protein concentrations as well as abundance of steroid acute regulatory protein, cytochrome P450 19A1, and multidrug resistance-associated protein 1 were not affected by HS (P > 0.05). HS increased estrogen sulfotransferase protein abundance (44%; P = 0.02), toll-like receptor 4 (36%; P = 0.05), and phosphorylated REL-associated protein (31%; P = 0.02). Regardless of treatment, toll-like receptor 4 protein was localized to mural granulosa cells in the porcine ovary. In conclusion, HS altered ovarian signaling in postpubertal gilts during their follicular phase in ways that likely contributes to seasonal infertility.

Chronic toxicity and endocrine disruption of naproxen in freshwater waterfleas and fish, and steroidogenic alteration using H295R cell assay

Naproxen is a non-steroidal anti-inflammatory drug (NSAID) and has been frequently detected in surface waters around the world. Although endocrine disruption and reproduction related effects of NSAIDs are of increasing concern, the ecotoxicity of naproxen in aquatic organisms is limited primarily to acute lethal effects. In this study, chronic toxicity of naproxen was evaluated employing two daphnids (Daphnia magna and Moina macrocopa) and a fish (Oryzias latipes). The effects of naproxen on sex steroid hormones and gene transcription related to steroidogenesis were also evaluated in H295R cells. The chronic no observed effect concentrations (NOECs) of naproxen for reproduction were determined to be 10 mg L−1 in D. magna and 0.3 mg L−1 in M. macrocopa. At concentrations of 0.5 mg L−1, the survival of juvenile medaka fish was significantly decreased and transcription of erβ2 gene was significantly increased. Concentration of 17β-estradiol (E2) and the ratio of E2 and testosterone were significantly increased in H295R cells at 10 mg L−1, suggesting that naproxen could modulate sex hormone production. The current detected levels of naproxen in ambient Korean rivers are far lower than the effective levels, however potential adverse effects cannot be ignored in some highly polluted areas. Endocrine disruption effects in fish warrant further investigation particularly for their ecological implications.

17β-Estradiol on the Expression of G-Protein Coupled Estrogen Receptor (GPER/GPR30) Mitophagy, and the PI3K/Akt Signaling Pathway in ATDC5 Chondrocytes In Vitro.

BackgroundOsteoarthritis is a progressive inflammatory joint disease resulting in damage to articular cartilage. G-protein coupled estrogen receptor (GPER/GPR30) activates cell signaling in response to 17β-estradiol, which can be blocked by the GPR30 agonist, G15, an analog of G-1. The aims of this study were to investigate the effects of 17β-estradiol on the expression of G-protein coupled estrogen receptor (GPER/GPR30) on mitophagy and the PI3K/Akt signaling pathway in ATDC5 chondrocytes in vitro.Material/MethodsCultured ATDC5 chondrocytes were treated with increasing concentrations of 17β-estradiol with and without G15, p38 inhibitor (SB203580), JNK inhibitor (SP600125), PI3K inhibitor (LY294002, S1737), and mTOR inhibitor (S1842). Expression of GPER/GPR30 and components of the PI3K/Akt pathway in cultured ATDC5 chondrocytes were detected by immunofluorescence (IF) staining, Western blot, and real-time polymerase chain reaction (RT-PCR). Transmission electron microscopy (TEM) and IF were used to detect mitophagosomes. Expression of LC-3, LAMP2, TOM20, Hsp60, p-Akt, p-mTOR, p-p38, and p-JNK was investigated by Western blot. Proliferation and viability of the ATDC5 chondrocytes were determined using BrdU and MTT assays.ResultsIn 17β-estradiol-treated ATDC5 chondrocytes, increased expression of GPER/GPR30 was found, but fewer mitophagosomes were observed, and decreased numbers of TOM20-positive granules were co-localized with decreased LAMP2 and increased expression levels of TOM20, Hsp60, p-Akt, and p-mTOR, and reduced expression of LC3-II, were found. In 17β-estradiol-treated ATDC5 chondrocytes, the proliferation and viability of the 17β-estradiol-treated ATDC5 chondrocytes were significantly elevated.ConclusionsTreatment with 17β-estradiol protected ATDC5 chondrocytes against mitophagy via the GPER/GPR30 and the PI3K/Akt signaling pathway.

Estimation of human-origin estrone and 17β-estradiol concentrations in the Han River, Seoul, South Korea and its uncertainty-based ecological risk characterization

Estrogens originated from humans can reach ambient water and possibly cause significant ecological risks. In this study, the quantities of human-origin estrone (E1) and 17β-estradiol (E2) in the influent and effluent of four sewer treatment plants (STPs) in Seoul, South Korea were estimated using a demographic model. A Monte Carlo simulation was used to assess the quantitative uncertainty of estimated E1 and E2 concentrations. Mean concentrations of E1 and E2 estimated for STP influents ranged from 29.5 to 38.4 and 7.5 to 9.7ng/L, respectively. Meanwhile, mean concentrations of E1 and E2 estimated for STP effluents were 4.9 to 6.6 and 0.28 to 0.36ng/L, respectively. These estimated values are similar to measured data as reported in the literatures within the range of uncertainty based on the Monte Carlo simulation. The hazard quotient (HQ) value in the main stem of the Han River was calculated to be far less than 1 because of the dilution effect of the Han River's abundant flow, indicating that most of the Han River ecosystem will not be influenced by these endogenous estrogens. With a 95% cumulative probability, HQ values in the main stem of the Han River for the Jungrang, Nanji, Tanchun, and Seonam STPs were less than 0.18, 0.07, 0.08, and 0.15, respectively. Nevertheless, HQ values >1 were observed in the vicinity of the STP outlets when using the numerical modeling. Our results show that the endocrine disruption potential of E1 and E2 around STP outlets in the main stem of the Han River must be monitored carefully.

Effect of Melatonin Implants during the Non-Breeding Season on the Onset of Ovarian Activity and the Plasma Prolactin in Dromedary Camel.

To examine a possible control of reproductive seasonality by melatonin, continual-release subcutaneous melatonin implants were inserted 4.5 months before the natural breeding season (October–April) into female camels (Melatonin-treated group). The animals were exposed to an artificial long photoperiod (16L:8D) for 41 days prior to implant placement to facilitate receptivity to the short-day signal that is expected with melatonin implants. The treated and control groups (untreated females) were maintained separately under outdoor natural conditions. Ovarian follicular development was monitored in both groups by transrectal ultrasonography and by plasma estradiol-17β concentrations performed weekly for 8 weeks and then for 14 weeks following implant insertion. Plasma prolactin concentrations were determined at 45 and 15 days before and 0, 14, 28, 56, and 98 days after implant insertion. Plasma melatonin concentration was determined to validate response to the artificial long photoperiod and to verify the pattern of release from the implants. Results showed that the artificial long photoperiod induced a melatonin secretion peak of significantly (P < 0.05) shorter duration (about 2.5 h). Melatonin release from the implants resulted in higher circulating plasma melatonin levels during daytime and nighttime which persisted for more than 12 weeks following implants insertion. Treatment with melatonin implants advanced the onset of follicular growth activity by 3.5 months compared to untreated animals. Plasma estradiol-17β increased gradually from the second week after the beginning of treatment to reach significantly (P < 0.01) higher concentrations (39.2 ± 6.2 to 46.4 ± 4.5 pg/ml) between the third and the fifth week post insertion of melatonin implants. Treatment with melatonin implants also induced a moderate, but significant (P < 0.05) suppressive effect on plasma prolactin concentration on the 28th day. These results demonstrate that photoperiod appears to be involved in dromedary reproductive seasonality. Melatonin implants may be a useful tool to manipulate seasonality and to improve reproductive performance in this species. Administration of subcutaneous melatonin implants during the transition period to the breeding season following an artificial signal of long photoperiod have the potential to advance the breeding season in camels by about 2.5 months.