Plasma Estradiol Concentrations Research Articles

Comparative effects of Bisphenol S and Bisphenol A on the development of female reproductive system in rats; a neonatal exposure study

Bisphenol A (BPA) has been well documented for its endocrine disrupting potential however, very little is known about endocrine disrupting abilities of bisphenol S (BPS). The present study aimed to compare the endocrine disrupting potentials of BPS with BPA, using female rats as an experimental animal model. On postnatal day 1 (PND 1) female pups born were randomly assigned to seven different treatments. Control group received subcutaneous injection of castor oil (50 μL) from PND 1 to PND 10. Three groups of female pups were injected subcutaneously with different concentrations (0.5, 5 and 50 mg/kg in 50 μL castor oil) of BPS, while remaining three groups were treated with 0.5, 5 and 50 mg/kg BPA. Highest doses treatments of both compounds resulted in delayed puberty onset and altered estrous cyclicity. Final body weight was significantly high in the highest dose treated groups of both BPS and BPA. Gonadosomatic index, absolute and relative weight of uteri was significantly reduced in BPS (5 and 50 mg/kg) and BPA (5 and 50 mg/kg) treated groups than control. Plasma concentrations of testosterone and estradiol were significantly increased, while plasma progesterone, Luteinizing hormone (LH) and Follicle stimulating hormone (FSH) concentrations were significantly reduced in highest doses treated groups. Dose dependent increase in the number of cystic follicles in the ovaries was evident along with an increase in the number of atratic follicles. The results suggest that neonatal exposure to higher concentrations of BPS can lead to BPA like structural and endocrine alterations in female rats.

Effect of superstimulation on the expression of microRNAs and genes involved in steroidogenesis and ovulation in Nelore cows

To better understand the impact of ovarian superstimulation on bovine follicular microenvironment, Nelore cows (Bos taurus indicus) were subjected to ovarian superstimulation with follicle stimulating hormone (FSH, n = 10; P-36 protocol) or FSH combined with eCG (n = 10; P-36/eCG protocol). Follicular fluid was analyzed for cholesterol concentration. Granulosa cells were analyzed by RT-qPCR to assess the expression of genes involved in steroidogenic and ovulatory and expression of microRNAs involved in final follicular development and luteinizing hormone/choriogonadotropin receptor (LHCGR) expression. Plasma concentration of estradiol was also measured. Follicular fluid from the P-36 group showed higher concentration of cholesterol than that of control (non-superstimulated) cows. Plasma concentration of estradiol was higher in the P-36/eCG group. Abundance of STAR and FSHR mRNAs were lower in granulosa cells from the P-36/eCG group. In contrast, LHCGR mRNA abundance was higher in superstimulated granulosa cells from the P-36 group and showed a pattern opposite to that of miR-222 expression. Ovarian superstimulation did not affect the expression of other markers (mmu-miR-202-5p, has-miR-873, has-miR-144, and their target genes, CREB, TGFBR2, and ATG7) of antral follicle development. However, the mRNA expression of VEGF pathway components was modulated by P-36 treatment. Taken together, these results demonstrate that superstimulatory protocols modify steroidogenic capacity, increase plasma estradiol, and regulate the abundance of VEGF system, LHCGR mRNA and suppress the expression of miR-222 in bovine granulosa cells.

Seasonal steroid variations in relation to maturity stages in the female Pacific red snapper Lutjanus peru in the Gulf of California, Mexico

Reproductive steroid concentration and gonadal development as well as plasma cortisol were investigated during the reproductive period (April to October) of the Pacific red snapper Lutjanus peru, a commercial and important local fishery species in the surrounding areas of Bahía La Paz, Gulf of California, Mexico. Gonads in April were immature and characterized by the presence of oocytes at the chromatin nucleolar and perinucleolar stages. In the next month, more than 50% of the fish showed gonads in vitellogenesis, characterized by the presence of cortical alveoli and early vitellogenic (V1) oocytes. Late vitellogenic and mature gonads were present in July, August and September. Postovulatory follicles were observed only in gonads collected in August, indicating spawning activity of these individuals. The gonadosomatic index (GSI) increased significantly (p <0.05) from April and May to July, when it reached its maximum value. A significant increase (p <0.05) in plasma testosterone (T) was also observed in July compared to previous months, and plasma estradiol (E2) concentration was significantly higher (p <0.05) in September than in April. Cortisol concentration was significantly higher (p <0.05) in September than in May. Significant differences (p <0.05) between gonadal stages were observed for GSI and E2 but not for T. We conclude that the Pacific red snapper, in the surrounding areas of Bahía de la Paz, showed an asynchronous ovarian growth starting in May when water temperature increased and that was characterized by several spawning events.

Inflammatory status and its relationships with different patterns of postpartum luteal activity and reproductive performance in early lactating Holstein cows

This study was undertaken to determine the effect of inflammatory status on different patterns of postpartum luteal activity and reproductive performance of Holstein cows during early lactation. The cows (n = 75) averaged 3.4 ± 1.2 (mean ± SEM) in parity and 3.1 ± 0.2 (mean ± SEM) in body condition score at calving. Transrectal ultrasonography was performed twice weekly from day 10–60 postpartum to consider ovarian dynamics. Plasma concentrations of progesterone (P4) and estradiol (E2) were measured twice weekly and plasma levels of tumor necrosis factor alpha (TNF-α) and lipopolysacchride binding protein (LBP) were measured weekly (week 3–6 postpartum). Based on plasma P4 and E2 results, 34 (45.3%) cows had normal luteal activity (NLA), whereas 19 (25.3%), 12 (16.0%), 7 (9.3%) and 3 (4.0%) cows had prolonged luteal phase (PLP), delayed first ovulation (DO), anovulation (AO) and short luteal phase (SLP), respectively. Plasma TNF-α and LBP concentrations were affected by postpartum luteal activity (NLA, SLP and PLP), ovulatory status (DO and AO) and number of weeks postpartum (P < .05). These concentrations were greater in cows with PLP (P < .05) as compared to NLA cows, and in cows that had delayed ovulation or anovulation compared to ovulated cows (P < .05). Cows with PLP had greater open days and lower conception rate as compared to NLA cows (P < .05). Healthy cows had a larger CL and greater plasma estradiol and progesterone concentrations at first and second cycle postpartum compared to inflamed cows and followed it with greater fertility (P < .05). In conclusion, inflammatory statuses were different in high-producing dairy cows showing PLP, AO and DO in comparison with the postpartum normal luteal activity (NLA) cows that influenced reproduction outcomes.

Molecular changes in premenopausal oestrogen receptor-positive primary breast cancer in Vietnamese women after oophorectomy

For premenopausal women with primary ER + breast cancer, oophorectomy (OvX) is an evidence-based cost-effective option and is standard treatment in many countries. However, there is virtually no data describing the effects of OvX on breast tumour biology. We therefore, characterised the endocrine and genome-wide transcriptional impact of OvX in 56 premenopausal women with ER + breast cancer for 2 weeks prior to mastectomy. Plasma estradiol concentrations decreased from 406 ± 41 to 20.7 ± 2.6 pmol/l (mean ± sem) 24 h after OvX, and to 8.1 ± 0.8 pmol/l 2 weeks later at mastectomy. Ki67 decreased in 33/36 (91.7%) tumours. The expression of 655 genes changed significantly (FDR < 1%) with an absolute mean fold-change (FC) ≥ 1.25 (257 up, 398 down). Archetypal oestrogen-regulated genes (TFF1, GREB1, PGR and PDZK1) showed large decreases in expression (FC = 0.20–0.69; p < 1e-6-1e-7). Proliferation-associated genes (e.g. TOP2A, AURKA and UBE2C) were also strongly downregulated (FC = 0.38–0.56; p < 1e-7) along with putative progesterone-regulated genes (e.g. FKBP4, MYB; FC = 0.64–0.68; p < 1e-4-1e-7). The gene expression changes did not differ according to HER2 status and correlated strongly with the changes reported previously after aromatase inhibitor (AI) treatment in postmenopausal women (rho = 0.55, p < 1e-04). However, after OvX the mean FC was significantly higher compared to AI (p < 1e-04). In conclusion, changes in tumoural gene expression after OvX were largely similar, but of a greater magnitude to those observed after AI in postmenopausal patients; however, OvX appeared to have a greater effect on progesterone-regulated genes than AI.

Expression of two glycoprotein hormone receptors in larval, parasitic phase, and adult sea lampreys

All jawed vertebrates have three canonical glycoprotein hormones (GpHs: luteinizing hormone, LH; follicle stimulating hormone, FSH; and thyroid stimulating hormone, TSH) with three corresponding GpH receptors (GpH-Rs: LH-R, FSH-R, and TSH-R). In contrast, we propose that the jawless vertebrate, the sea lamprey (Petromyzon marinus), only has two pituitary glycoprotein hormones, lamprey (l)GpH and l-thyrostimulin, and two functional glycoprotein receptors, lGpH-R I and II. It is not known at this time whether there is a specific receptor for lGpH and l-thyrostimulin, or if both GpHs can differentially activate the lGpH-Rs. In this report, we determined the RNA expression of lGpH-R I and II in the gonads and thyroids of larval, parasitic phase, and adult lampreys. A highly sensitive dual-label fluorescent in situ hybridization technique (RNAScope™) showed lGpH-R I expression in the ovaries of larval lamprey, and co-localization and co-expression of lGpH-R I and II in the ovaries of parasitic phase and adult lampreys. Both receptors were also highly co-localized and co-expressed in the endostyle of larval lamprey and thyroid follicles of parasitic and adult lampreys. In addition, we performed in vivo studies to determine the actions of lamprey gonadotropin releasing hormones (lGnRHs) on lGpH-R I and II expression by real time PCR, and determined plasma concentrations of estradiol and thyroxine. Administration of lGnRH-III significantly (p ≤ 0.01) increased lGpHR II expression in the thyroid follicles of adult female lampreys but did not cause a significant increase in RNA expression of lGpH-R I and II in ovaries. Concomitantly, there was a significant increase (p ≤ 0.01) of plasma estradiol without any significant changes of plasma thyroxine concentrations in response to treatment to lGnRH-I, -II, or -III. In summary, our results provide supporting evidence that the lamprey pituitary glycoprotein hormones may differentially activate the lamprey GpH-Rs in regulating both thyroid and gonadal activities during each of the three life stages of the sea lamprey.

Determinants of prolactin in postmenopausal Chinese women in Singapore.

Mechanistic and observational data together support a role for prolactin in breast cancer development. Determinants of prolactin in Asian populations have not been meaningfully explored, despite the lower risk of breast cancer in Asian populations. Determinants of plasma prolactin were evaluated in 442 postmenopausal women enrolled in the Singapore Chinese Health Study, a population-based prospective cohort study. At baseline all cohort members completed an in-person interview that elicited information on diet, menstrual and reproductive history, and lifestyle factors. One year after cohort initiation we began collecting blood samples. Quantified were plasma concentrations of prolactin, estrone, estradiol, testosterone, androstenedione, and sex hormone-binding globulin (SHBG). Analysis of covariance method was used for statistical analyses with age at blood draw, time since last meal, and time at blood draw as covariates. Mean prolactin levels were 25.1% lower with older age at menarche (p value = 0.001), and 27.6% higher with greater years between menarche and menopause (p value = 0.009). Prolactin levels were also positively associated with increased sleep duration (p value = 0.005). The independent determinants of prolactin were years from menarche to menopause, hours of sleep, and the plasma hormones estrone and SHBG (all p values < 0.01). The role of prolactin in breast cancer development may involve reproductive and lifestyle factors, such as a longer duration of menstrual cycling and sleep patterns.

Uterine endometrial vascularization during ovarian follicular growth in llamas: The effect of estradiol plasma concentration

The aim of this study was to evaluated changes in endometrial vascularization area (EVA) between the left and right uterine horn: a) during the ovarian follicular growth in intact llamas, and b) after exogenous estradiol administration of estradiol benzoate in ovariectomized (OVX) llamas. In experiment 1 follicle wave emergence was synchronized (n = 5 llamas) by follicle ablation (Day 0). Females were examined every other day from Day 1 to Day 27, using B mode ultrasonography to evaluate dominant follicle growth profile. Also, EVA was evaluated in each horn using Color-Power Doppler. Blood samples were taken every other day from Day 1 to Day 27 to measure estradiol (E2) plasma concentration by RIA. In experiment 2 OVX llamas (n = 4 llamas/group) were given a single im administration of: a) 1 mg of estradiol benzoate (EB) or b) 1 mL of saline. Females were subjected to ultrasound examinations every 48 h from Day −4 until treatment (Day 0), every 12 h from Day 0 to Day 4, and again every 48 h from Day 5 to Day 11. Evaluation of EVA in both uterine horns was performed as described for experiment 1. Blood sampling for the measurement of E2 was carried out at the same time points indicated for the ultrasound examinations. Serial data were analyzed by one way ANOVA for repeated measures using the MIXED Procedure in SAS. Also, Pearson's correlation was used to determine the relationship between variables. In intact llamas there was an effect of day on the dominant follicle growing profile (P < 0.01) and estradiol plasma concentration (P < 0.05). Dominant follicle diameter positively correlated (r = 0.4; P < 0.017) with estradiol plasma concentration. Also, EVA of right and left uterine horn did not differ (P = 0.89) during the evaluation period; however, it was affected by time (P < 0.05). In ovariectomized llamas estradiol concentration was significantly (P < 0.001) affected by treatment, time and their interaction. Accordingly, treatment with EB (P < 0.0001), time (P < 0.05) and their interaction (P < 0.01) affected EVA of both uterine horns; however, this variable did not differ between horns (P = 0.98). In conclusion, circulating concentrations of estradiol determined an increase in uterine vascularization, during the phase of follicular growth in intact llamas and after the exogenous administration of EB to ovariectomized females; however, no differential effect in endometrial vascularization area between right and left uterine horn was observed.

Supplementation of dairy cows with docosahexaenoic acid did not affect ovarian activity

The effect of docosahexaenoic acid (DHA) on ovarian activity of dairy cows was determined. Experimental cows (n = 25) were fed a total mixed ration supplemented daily with 100 g/cow of an algae product All-G-Rich (Alltech, Ireland) containing 10% DHA divided into 2 doses for 52 days. Determination of DHA from milk samples taken from all cows was performed before the All-G-Rich supplementation (on Day 0, D0), and on D21 and D42 of algae supplementation. Cows were synchronized to be in oestrus on D21 and D42 of the experimental period. Monitoring of ovarian activity was performed by transrectal ultrasonography. Examinations were performed at 2–3-day intervals from D0 until D52 of the experimental period. Plasma concentrations of progesterone, oestradiol, insulin, NEFA, and cholesterol were determined. Control cows (n = 25) were examined in the same way as the experimental cows. Milk DHA concentrations on D21 and D42 were significantly higher in treated cows (D21 1.38 vs 0.28, P &amp;lt; 0.0001; D42 1.34 vs 0.20, P &amp;lt; 0.0001). There were neither important effects of DHA on ovarian structures, nor on evaluated variables in plasma. Cows in the experimental group tended to have larger corpora lutea and higher cholesterol concentrations, but differences were not significant.

Sauna exposure immediately prior to short-term heat acclimation accelerates phenotypic adaptation in females

ObjectivesInvestigate whether a sauna exposure prior to short-term heat acclimation (HA) accelerates phenotypic adaptation in females. DesignRandomised, repeated measures, cross-over trial. MethodsNine females performed two 5-d HA interventions (controlled hyperthermia Tre≥38.5°C), separated by 7-wk, during the follicular phase of the menstrual cycle confirmed by plasma concentrations of 17-β estradiol and progesterone. Prior to each 90-min HA session participants sat for 20-min in either a temperate environment (20°C, 40% RH; HAtemp) wearing shorts and sports bra or a hot environment (50°C, 30% RH) wearing a sauna suit to replicate sauna conditions (HAsauna). Participants performed a running heat tolerance test (RHTT) 24-h pre and 24-h post HA. ResultsMean heart rate (HR) (85±4 vs. 68±5 bpm, p≤0.001), sweat rate (0.4±0.2 vs. 0.0±0.0Lh−1, p≤0.001), and thermal sensation (6±0 vs. 5±1, p=0.050) were higher during the sauna compared to temperate exposure. Resting rectal temperature (Tre) (−0.28±0.16°C), peak Tre (−0.42±0.22°C), resting HR (−10±4 bpm), peak HR (−12±7 bpm), Tre at sweating onset (−0.29±0.17°C) (p≤0.001), thermal sensation (−0.5±0.5; p=0.002), and perceived exertion (−3±2; p≤0.001) reduced during the RHTT, following HAsauna; but not HAtemp. Plasma volume expansion was greater following HAsauna (HAsauna, 9±7%; HAtemp, 1±5%; p=0.013). Sweat rate (p≤0.001) increased and sweat NaCl (p=0.006) reduced during the RHTT following HAsauna and HAtemp. ConclusionsThis novel strategy initiated HA with an attenuation of thermoregulatory, cardiovascular, and perceptual strain in females due to a measurably greater strain in the sauna compared to temperate exposure when adopted prior to STHA.

Variable aromatase inhibitor plasma concentrations do not correlate with circulating estrogen concentrations in post-menopausal breast cancer patients.

The aromatase inhibitors (AI) exemestane (EXE), letrozole (LET), and anastrozole suppress estrogen biosynthesis, and are effective treatments for estrogen receptor (ER)-positive breast cancer. Prior work suggests that anastrozole blood concentrations are associated with the magnitude of estrogen suppression. The objective of this study was to determine whether the magnitude of estrogen suppression, as determined by plasma estradiol (E2) concentrations, in EXE or LET treated patients is associated with plasma AI concentrations. Five hundred post-menopausal women with ER-positive breast cancer were enrolled in the prospective Exemestane and Letrozole Pharmacogenetic (ELPh) Study conducted by the COnsortium on BReast cancer phArmacogomics (COBRA) and randomly assigned to either drug. Estrogen concentrations were measured at baseline and after 3months of AI treatment and drug concentrations were measured after 1 or 3months. EXE or LET concentrations were compared with 3-month E2 concentration or the change from baseline to 3months using several complementary statistical procedures. Four-hundred patients with on-treatment E2 and AI concentrations were evaluable (EXE n=200, LET n=200). Thirty (7.6%) patients (EXE n=13, LET n=17) had 3-month E2 concentrations above the lower limit of quantification (LLOQ) (median: 4.75; range: 1.42-63.8pg/mL). EXE and LET concentrations were not associated with on-treatment E2 concentrations or changes in E2 concentrations from baseline (all p>0.05). Steady-state plasma AI concentrations do not explain variability in E2 suppression in post-menopausal women receiving EXE or LET therapy, in contrast with prior evidence in anastrozole treated patients.

A diet supplemented with n-3 polyunsaturated fatty acids influences the metabomscic and endocrine response of rabbit does and their offspring.

The aim of the present study was to evaluate the productive, endocrine, and metabomscic responses as well as oxidative stress of rabbit does and their offspring when fed a diet supplemented with -3 PUFA during their first productive cycle. To this aim, a total of 105 rabbit does were fed ad mscibitum from d 60 to 172 of age 2 isoenergetic and isoproteic diets differing in fatty acid composition. The control diet ( = 52 does) contained 45.9 g/kg of -3 of the total fatty acids and the enriched diet ( = 53 does) contained 149.2 g/kg of -3 of the total fatty acids. Both experimental groups had similar feed intake during rearing, pregnancy, and lactation. The enrichment of diet had no effect on ultrasonographic assessment of does on d 9 and 16 of pregnancy, with an embryonic vesicle number and fetus and placenta size similar between groups ( > 0.05). Even though there were no major effects ( > 0.05) on fertimscity, duration of gestation, and number born amscive and stillborn kits at parturition, mscive kits from enriched does were longer (71.6 ± 2.42 vs. 79.5 ± 2.13 mm; < 0.05) and tended to be heavier (42.5 ± 3.94 vs. 50.8 ± 3.47 g; = 0.07) than those from control does ( < 0.05). The 2 groups had similar milk production and mortamscity values during lactation; consequently, there were no differences between diets in ADG, mscitter weight, and number of weaned kits ( > 0.05). In enriched does, higher plasma leptin and estradiol concentrations than in control does ( < 0.05) were observed. In addition, enriched females also had lower total and high-density mscipoprotein cholesterol (HDL-c) than control females during lactation ( < 0.05). Regarding offspring, the enrichment of diet with PUFA caused a hypermscipidemic status (greater values of plasma triglycerides, total cholesterol, and HDL-c; < 0.05) at 1 d postpartum (dpp), compared with the control group, that disappeared at 32 dpp. Supplemented does before parturition and their offspring at 1 dpp had greater oxidative stress than those in the control group. In conclusion, an increase of -3 PUFA concentration in the diet of rabbit does and, consequently, of their offspring during a productive cycle alters their mscipid profile and the indicators of oxidative stress, without major endocrine modifications or improvements in the productive variables.

Angiotensin II/Angiotensin (1-7) ratio and 24-h blood pressure throughout the menstrual cycle and in women using oral contraceptives.

Women using oral contraceptives have higher ambulatory blood pressures (BPs) than other women. We sought to learn whether this was associated with an alteration in the balance of angiotensin II (Ang)/Ang (1-7) and whether this ratio and BP remained constant throughout the menstrual cycle. In total, 30 (15 ovulatory, 15 taking oral contraceptives) healthy, normotensive women aged 18-30 years were studied. The ovulatory group was assessed within days 1-5 (follicular phase) and 19-23 (luteal phase) and the oral contraceptive group within days 19-23. Peripheral, central and 24-h BP, vascular wall stiffness, and body composition were measured along with plasma estradiol, progesterone, renin, aldosterone, Ang II, and Ang (1-7) concentrations. In ovulatory women plasma renin activity (P < 0.001), renin concentration (P < 0.01) and aldosterone (P < 0.05) were higher in the luteal than follicular phase, whereas BP, Ang II and the Ang II/Ang (1-7) ratio remained constant. In women taking oral contraceptives, plasma renin activity (P < 0.001) and concentration (P < 0.01) were higher than in follicular phase ovulatory women whereas 24-h BP, Ang II, Ang (1-7), and the Ang II/Ang (1-7) ratio (P < 0.01) were higher than in both phases of the ovulatory group. However, there was no significant correlation between BP and the Ang II/Ang (1-7) ratio. This study confirms that BP is constant throughout the normal menstrual cycle along with a constant balance between the vasoconstrictor (Ang II) and vasodilator [Ang (1-7)] arms of the renin-Ang-aldosterone system. Women taking oral contraceptives have a higher Ang II/Ang (1-7) ratio associated with their BP elevation although no causal relationship has been found.

A low-fat diet enriched in fish oil increased lipogenesis and fetal outcome of C57BL/6 mice.

There is clear evidence that nutritional strategy employed during pregnancy has profound influence on the offspring health outcomes. However, the effect of the quality and the quantity of maternal fat intake on maternal metabolic profile during different stages of pregnancy and its impact on pregnancy sustainability is not known. Female C57BL/6 mice (7 weeks old) were fed diets varying in the quantity of fat (5% vs 11%) for two weeks prior to mating and throughout pregnancy. The 5% fat diet was enriched with longer chain omega (n)-3 polyunsaturated fatty acids (PUFA) from fish oil. Maternal plasma and tissues were collected before mating and during pregnancy at days 6.5, 12.5 and 18.5. Plasma lipids, glucose, insulin, progesterone and estradiol levels were measured. Cholesterol efflux capacity of maternal plasma as well as the mRNA expression of placental steroidogenic acute regulatory protein and hepatic lipogenic genes (acetyl-CoA carboxylase-1, fatty acid synthase, diacylglycerol acyltransferase-2 and stearoyl-CoA desaturase-1) was determined. Feto-placental weight and fetuses sustained throughout gestation were recorded. A low-fat maternal diet enriched with n-3 PUFA increased maternal plasma triacylglycerol and the mRNA expression of rate-limiting lipogenic enzymes, along with increasing cholesterol efflux capacity (P < 0.05), likely to meet fetal lipid demand during pregnancy. Furthermore, diet enriched with longer chain n-3 PUFA increased the maternal plasma concentration of progesterone and estradiol during pregnancy (P < 0.05), which coincides with an increase in the number of fetuses sustained till day 18.5. These novel findings may be important when designing dietary strategies to optimize reproductive capability and pregnancy outcomes.

Characterization of the variability and repeatability of gonadotropin-releasing hormone-induced luteinizing hormone responses in dairy cows within a synchronized ovulation protocol.

The primary objective was to determine the variability and repeatability of GnRH-induced LH responses. The secondary objective was to evaluate the associations among plasma LH, FSH, estradiol (E2), and progesterone (P4) concentrations. One hundred lactating Holstein cows (35 primiparous, 65 multiparous) were initially subjected to a presynchronization protocol (d 0, PGF2α; d 3, GnRH) followed 7 d later by Ovsynch (d 10, GnRH; d 17, PGF2α; 56 h later, GnRH) and timed artificial insemination 16 h after the last GnRH. Blood samples were collected immediately before the GnRH injection of presynchronization and the second GnRH of Ovsynch to determine plasma concentrations of LH, FSH, and P4. A second blood sample was collected 2 h after each of the above GnRH injections to determine GnRH-induced LH and FSH concentrations. Plasma concentrations of E2 were also determined in samples collected immediately before the second GnRH of Ovsynch. Cows that (1) had higher LH concentrations at 0 h than at 2 h after GnRH, (2) showed an ongoing spontaneous LH surge, (3) did not respond to GnRH, and (4) had P4 ≥ 0.5 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch were excluded from the analysis. The variability (coefficient of variation) and repeatability [between animal variance/(within animal variance + between animal variance)] of GnRH-induced LH response were determined from samples collected 2 h after the GnRH of presynchronization and the second GnRH of Ovsynch. The associations among plasma LH, FSH, E2, and P4 were determined at the second GnRH of Ovsynch. Mean (±SEM) LH concentrations before GnRH were 0.5 ± 0.04 and 0.6 ± 0.03 ng/mL, whereas mean LH concentrations 2 h after GnRH were 9.8 ± 1.0 and 12.1 ± 0.8 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The variability of GnRH-induced LH was 76.1 and 52.1% at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The repeatability estimate for GnRH-induced LH concentration between GnRH of presynchronization and Ovsynch assessments was 0.10. Plasma concentrations of LH were positively associated with FSH and E2 (r = 0.61 and 0.30, respectively) and negatively associated with P4 (r = -0.46) at the second GnRH of Ovsynch. In summary, GnRH-induced LH responses were highly variable and unrepeatable, and LH concentrations were positively associated with FSH and E2 and negatively associated with P4.

Letrozole, an aromatase inhibitor, reduces post-peak age-related regression of rooster reproductive performance

This study was designed to evaluate orally administrated Letrozole (Lz) on reproductive performance, plasma testosterone and estradiol concentrations and relative abundance of mRNA of GnRH, FSH and LH in roosters. Ross 308 roosters (n=32) that were 40-weeks of age were individually housed and received a basal standard diet supplemented different amounts of capsulated Lz [0 (Lz-0), 0.5 (Lz-0.5), 1 (Lz-1) or 1.5 (Lz-1.5), mg Lz/bird/day] for 12 weeks. Sperm quality variables and plasma testosterone and estradiol concentrations were assessed from the first to the tenth week of the treatment period. Semen samples from the 11th to 12th week were used for artificial insemination and eggs were collected and allotted to assess fertility and hatchability rates. Relative abundance of hypothalamic and pituitary GnRH, LH and FSH mRNA was evaluated at the end of 12th week. The results indicated that total and forward sperm motility as well as egg hatchability rate were greater in the Lz-0.5 group. Greater sperm concentrations, ejaculate volume, sperm plasma membrane integrity, testis index and fertility rates were recorded for both Lz-0.5 and Lz-1 groups compared with the Lz-0 group (P<0.05). Body weight, percentage of sperm abnormalities, and sperm plasma membrane functionality were not affected by treatment. Testosterone and estradiol concentrations were negatively related with greater testosterone concentrations in the Lz-1.5 group which had lesser estradiol concentrations. Relative mRNA transcript abundance for GnRH, LH and FSH was Lz dose responsive being greater in the treated groups; however, this trend plateaued for GnRH and for the relative abundance of both LH and FSH mRNA was less in the Lz-1.5 group than the other treatment groups. It is concluded that Lz may be an effective treatment to improve age related post-peak reproductive performance of roosters.

Is incomplete estradiol suppression during aromatase inhibitor treatment in post-menopausal patients with breast cancer due to insufficient systemic drug concentrations?

1063 Background: Aromatase inhibitors (AI) suppress estrogen biosynthesis and are effective treatments for estrogen receptor (ER)-positive breast cancer. In a prospectively enrolled cohort we observed a subset of post-menopausal women who exhibit high plasma estradiol (E2) concentrations during AI treatment, which could potentially contribute to treatment failure. We tested the hypothesis that incomplete E2 suppression is due to insufficient systemic AI concentrations. Methods: Five hundred post-menopausal women with ER-positive breast cancer were randomized to daily exemestane (Exe) 25 mg or letrozole (Let) 2.5 mg. Plasma E2 was measured using GC/MS/MS (lower limit of quantification (LLOQ) = 1.25 pg/mL) at baseline and after 3 months. Let and Exe plasma concentrations measured after 1 or 3 months were compared with the magnitude of E2 depletion using four complementary statistical procedures to assess associations of drug concentrations with: 1) a binary outcome of E2 suppression below LLOQ (logistic regression), 2) 3-month E2 concentrations (linear regression), 3) absolute change from baseline in E2 concentrations (Spearman correlation), and 4) an ordinal outcome defined by E2: decreased to below LLOQ, decreased but not to LLOQ, stayed the same, or increased from baseline (cumulative logistic regression). Results: 397 patients with E2 and AI concentration measurements were evaluable (Exe n = 199, Let n = 198). Thirty (7.6%) patients (Exe n = 13, Let n = 17) had E2 concentrations above the LLOQ at 3 months (range: 1.42-63.8 pg/mL). Exe and Let concentrations were not associated with achievement of unmeasurable E2 concentrations, on-treatment E2 concentrations, E2 change from baseline, or ordinal groupings of E2 change (all p &gt; 0.05). In a parallel analysis there was no association of estrone-sulfate and drug concentrations (data not shown). Conclusions: Our results suggest that circulating drug concentrations do not explain incomplete E2 suppression in women receiving AI therapy. Additional studies are underway to determine whether age, body mass and genetic variation in the aromatase enzyme influence AI treatment response.

Effect of growth hormone on steroid concentrations and mRNA expression of their receptor, and selected egg-specific protein genes in the chicken oviduct during pause in laying induced by fasting

This study was undertaken to examine the effect of growth hormone (GH) treatment during pause in laying on (1) the concentration of steroids in blood plasma and oviduct tissues, (2) the expression of mRNA of steroid receptors, and (3) the mRNA expression of selected egg-specific proteins in the chicken oviduct. A pause in egg laying was induced by food deprivation for 5 d, followed by feeding every other day, and then feeding daily from Day 10 onward. Birds were divided into three groups: control (n = 18) fed ad libitum, subjected to pause in laying (n = 18), and subjected to pause in laying and injected every day with 200 μg/kg BW of chicken GH (chGH; n = 18). The oviduct was isolated from hens of each group on Days 6 (when the oviduct was regressed), 13 (during oviduct recrudescence), and 17 or 20 (rejuvenated oviduct) of the experiment. Fasting caused a decrease in plasma concentrations of progesterone (P4), testosterone, and estradiol on Day 6 and a reduction in tissue concentrations of these steroids on Days 6 and 13. Fasting also caused an increased relative expression of estrogen receptor α and β (ERα, ERβ) and progesterone receptor (PR) in the magnum and shell gland on Day 6, increased ERα and PR in the magnum on Days 13 and 17 or 20, and increased androgen receptor (AR) mRNA in the magnum on Days 6 and 13 and in the shell gland on Day 13. A fasting-induced elevation in ovocalyxin-36 mRNA expression on Day 6 and a decrease in avidin mRNA on Days 6 and 13 and in ovocleidin-116 on Day 13 were also observed (P < 0.05 to P < 0.001). Administration of chGH abolished the fasting-induced decrease in the concentration of steroids in plasma and tissues. Furthermore, chGH enhanced the effect of fasting on mRNA expression of PR, ERα, and avidin in the magnum on Day 6, and ERα in the shell gland on Day 13. The gene expression of ovalbumin on Days 6 and 13, ovocalyxin-36 and ovocleidin-116 on Day 6 was decreased in chGH-treated chickens. In contrast, the expression of ovalbumin on Day 17 or 20 was increased (P < 0.05 to P < 0.001). The results obtained indicate that, by alterations in the concentration of steroid hormones and their receptor expression in the chicken oviduct, GH determines the rate of regression and rejuvenation of this organ during molting. Moreover, changes in the expression of selected egg proteins indicate that GH might be the regulator of the secretory activity of the hen oviduct.

Effects of endurance training and herb supplementation on tissue nesfatin-1/nucleobindin-2 and ghrelin mRNA expression

This study investigated the effects of aerobic training and extract of Pistacia-atlantica (Baneh) as a plant with rich fatty acids, on tissue nesfatin-1/nucleobindin-2 and ghrelin gene expression, plasma high- and low-density lipoprotein, tissue/plasma total cholesterol and triglyceride concentrations in female rats. 20 Wistar rats were randomly assigned to saline-control, saline-training, Baneh-control, and Baneh-training groups. The training groups were given a specific exercise on a motor-driven treadmill for 8 weeks. Nesfatin-1/nucleobindin-2 and ghrelin mRNA expressions were detected by Real-time PCR method. According to the results, the trained rats had higher nesfatin-1 mRNA expressions in liver and visceral fat and liver estradiol concentration but their liver ghrelin mRNA expressions were lower compared to the control rats. Baneh-treated rats had lower nesfatin-1 mRNA expressions in liver and visceral fat and plasma HDL-C concentration, but their plasma estradiol concentrations were significantly higher. Moreover, liver estradiol concentration and liver ghrelin mRNA expression were negatively correlated, whereas the correlation between plasma estradiol and HDL-C concentrations and nesfatin-1 mRNA expression in visceral fat was significantly positive. Generally, exercise and a high-fat diet induced changes in liver ghrelin, nesfatin-1 mRNA expression, liver estradiol level, and nesfatin-1 in visceral fat, nesfatin-1 expression in liver and visceral fat, liver glucose, plasma HDL-C, and estradiol.

Detrimental effect of maternal and post-weaning high-fat diet on the reproductive function in the adult female offspring rat: roles of insulin-like growth factor 2 and the ovarian circadian clock

PurposeWe evaluate the impact of maternal and post-weaning high-fat (HF) diet on ovarian follicular population, steroidogenesis, and gene expression with a focus on the circadian clock system and insulin-like growth factor 2 (Igf2) in adult offspring ovaries, and to elucidate whether a maternal and post-weaning diet confers similar risks.MethodsVirgin Sprague-Dawley rats were fed with normal chow (C) diet or HF diet for 5 weeks before mating, during gestation, and lactation. Female offspring were fed with the C or HF diet from weaning to 6 months of age, resulting in four study groups (n = 6 per group): C/C, C/HF, HF/C, and HF/HF.ResultsOvaries from offspring exposed to post-weaning HF diet (i.e., the C/HF and HF/HF groups) had a decrease in small follicle numbers, but with similar numbers of antral follicles and corpora lutea. Offspring from HF-fed dams (i.e., the HF/C and HF/HF groups) had increased plasma estradiol concentrations and decreased luteinizing hormone levels at 6 months of age. In addition, Igf2 and each of the circadian rhythm core genes Clock, Per1, Per2, and Per3 were increased in the ovaries of offspring exposed to maternal HF diet (both HF/C and HF/HF groups).ConclusionsMaternal and post-weaning HF diet programs the reproductive profile of the female offspring in adult life through different manners. Post-weaning HF intake resulted in the reduction of small follicles in adulthood, whereas maternal HF diet had long-term deleterious consequences on female offspring steroidogenesis and coincided with alteration of the upregulation of the imprinted gene Igf2 and changes in ovarian circadian rhythms.