Progesterone Concentrations Research Articles

THE PROGINS VARIANT OF THE PGR GENE AND PLACENTAL ENDOCRINE FUNCTION AT RISK OF PRETERM LABOR: A PILOT STUDY

Progesterone receptors, which are encoded by the PGR gene, mediate the main physiological effects of progesterone – the hormone that is required for a successful pregnancy. The PROGINS variant is one of the most common variants of the PGR gene. Due to genetic heterogeneity, the presence of inferior forms of progesterone receptors in women can be a clinical problem. In addition to fertility problems, such women may theoretically respond differently to progesterone replacement therapy.Aim of the study to determine the effect of the PROGINS variant of the PGR gene on the hormonal status and results of progesterone deficiency treatment in pregnant women at risk of preterm labor.Material and methods. The main group included 30 pregnant women with progesterone deficiency and the threat of preterm labor, and the comparison group included 30 pregnant women with a normal course of pregnancy. The concentrations of progesterone, estradiol, and placental lactogen were measured in the pregnant women of both groups. Genotyping was assayed by PCR.Results. The genotypic frequencies of the PROGINS variant were not significantlydifferent between the main and comparison groups. Pregnant patients in the main group with the T2T2 genotype after treatment had significantly lower progesterone levels compared to patients with the T1T1 and T1T2 genotypes. Pregnant women in the main group with the T2T2 genotype delivered at a shorter gestational age.Conclusion. The results of our study showed the associations between the PROGINS variant of the PGR gene and the inefficacy of progesterone deficiency treatment in pregnant women at risk of preterm birth and shortened gestation.

Plasma progesterone profiles in ewes using different injectable progesterone formulations

Context The use of progesterone vaginal implants in sheep is frequently associated with vaginitis and adherences; and can hamper fertility in estrus synchronization programs. Aims The aim of this study was to characterize the plasma hormone profile induced by two injectable progesterone (iP4) formulations in ewes. Methods In Experiment 1, ewes received 20 or 40 mg of an iP4, with a third group as a control. In Experiment 2, 75 or 150 mg of a long-acting iP4 were administered, again with a control group. Key results In Experiment 1, progesterone concentrations remained at luteal levels for less than 24 h. The intervals from iP4 treatment to the onset of estrus and ovulation were greater in treated than in control ewes (P < 0.01). The proportion of ewes that ovulated within 8 days after treatment was lower in ewes that received 40 mg iP4 than in control ewes (P < 0.05). In Experiment 2, mean progesterone concentrations remained at luteal levels for 120 h. The interval from iP4 administration to estrus was longer in 150 mg treated ewes than in control ewes (P < 0.01). The interval from treatment to ovulation was longer in ewes treated with 150 mg than 75 mg and control ewes (P < 0.05). The proportion of ewes that ovulated was lower at 150 mg (P < 0.05). Conclusions The iP4 formula used in Experiment 1 could not maintain luteal levels for more than 24 h. The use of long-acting iP4 maintained plasma progesterone concentrations above luteal levels for at least 5 days. The long-acting treatment can therefore be tested for estrous synchronization treatments. Implications The long-acting progesterone may be an alternative to short estrous synchronization protocols.

Support of the luteal phase in programs for transferring thawed embryos into the uterine cavity in patients with overweight and obesity

Introduction. Progesterone levels are a critical factor for embryo implantation in in vitro fertilization (IVF) programs, including thawed embryo transfer programs. Overweight/obese patients often have lower blood progesterone concentrations in the luteal phase during IVF cycles, which requires an individual approach to planning luteal support in this category of patients.Aim. To compare the pregnancy ratе and miscarriage rates up to 12 weeks of pregnancy in overweight/obese patients who underwent a program of transfer of thawed embryos into the uterine cavity, depending on the type and route of administration of gestagens.Materials and methods. The study included 76 overweight or obese patients. All patients were treated for infertility using a program for transferring a thawed embryo into the uterine cavity; luteal phase support was carried out either using micronized progesterone, identical to natural, – the drug Utrogestan in a daily dose of 800 mg intravaginally (40 patients), or combination therapy was prescribed – vaginal gel with progesterone 90 mg in combination with the synthetic progestin dydrogesterone in a daily dose of 30 mg, taken orally (36 patients).Results. The rates of pregnancy and miscarriage in the first trimester were comparable in both groups and did not depend on differences in luteal support.Conclusions. The formation of the luteal phase of the cycle in patients with overweight/obesity is equally effective using various forms of gestagens. The use of micronized progesterone may be preferable in patients with overweight/obesity in terms of the ability to monitor blood progesterone levels, a more favorable metabolic profile when used vaginally, and the possibility of switching to oral administration of the drug in the event of the development of adverse local events.

Perceived Negative Menstrual Cycle Symptoms, but not Changes in Estrogen or Progesterone, are Associated with Impaired Cycling Race Performance.

Purpose: To examine the relationship between menstrual cycle (MC) phase-dependent fluctuations of estrogen and progesterone and virtual cycling race performance, with a secondary aim of correlating perceived MC-related symptoms with performance.Methods: In a novel observational study design, thirty-seven female cyclists/triathletes not using any hormonal contraception completed one virtual cycling race [19.5 km time trial (TT)] per week across a one-month period (totaling four races). Participants completed MC characterization and tracking, including urinary ovulation kits, across two complete MCs. Venous blood samples were collected within 21 h of racing to determine serum 17-β-estradiol and progesterone concentrations, as well as an assessment of self-reported, perceived race-day MC and gastrointestinal (GI) symptoms, which were all then correlated to race performance.Results: There was no relationship between race completion time and individual estradiol (r = -0.001, p = 0.992) or progesterone (r = -0.023, p = 0.833) concentrations. There was no difference between race time between MC phases (follicular/luteal, p = 0.238), whether MC bleeding or not bleeding (p = 0.619) and ovulating or not ovulating (p = 0.423). The total number of perceived MC symptoms recorded on race day was positively correlated to increased race time [r = 0.268 (95% CI 0.056 to 0.457), p = 0.014], as was the number of GI symptoms of at least "moderate" severity before the race (r = 0.233 [95% CI 0.021 to 0.425], p = 0.031), but not post-race (r = 0.022, p = 0.841).Conclusions: When implementing a novel, virtual cycling race, fluctuations in ovarian hormone concentrations across the MC do not appear to affect real-world cycling performance among trained cyclists, while perceived negative MC and GI symptoms may relate to impaired performance. Therefore, the management of negative MC and GI symptoms appears important for athletic performance enhancement or to mitigate performance decline.

Progesterone peak influences embryonic developmental morphokinetics on trigger day? A retrospective study

ObjectivePremature Progesterone Rise (PPR) is characterized by elevated serum progesterone concentrations either towards the end of the follicular phase or on the trigger day, surpassing a pre-defined threshold value. Aim of the study is to evaluate the impact of PPR exceeding 1.5 ng/ml at the time of hCG-trigger on embryo morphokinetic parameters and to identify predictive biomarkers of in IntraCytoplasmic Sperm Injection (ICSI) cycles outcomes.MethodsIt is a retrospective study including patients underwent ICSI cycles in the period 2020–2023. 58 patients were recruited in the study group showing P levels in the trigger day greater than or equal to 1.5 ng/ml. A matching control group of 58 patients with P levels below 1.5 ng/ml was after selected. The general characteristics of these patients, including age, Body Mass Index (BMI), antral follicle count (AFC), anti-Müllerian hormone (AMH) and follicle-stimulating hormone (FSH) levels, the type of infertility and smoking/non-smoking patients, were recorded on the day of their initial visit. Subsequently, data were collected regarding the number of eggs retrieved, mature eggs, successfully fertilized eggs, and embryos reaching the blastocyst stage. Additionally, the timing of embryonic development and the quality of obtained blastocysts, as assessed by the degree of expansion and the characteristics of the inner cell mass (ICM) and trophectoderm (TE), were evaluated using Time-Lapse technology.ResultsElevated P levels exceeding 1.5 ng/ml on the trigger day were directly associated with a significantly larger number of antral follicles, consequently leading to a higher count of retrieved eggs, mature eggs, successfully fertilized eggs and embryos reaching the blastocyst stage. Furthermore, the analysis of morphokinetic parameters indicated faster division times and a notably greater number of high-grade blastocysts in the study group compared to the control group.ConclusionsP levels ≥ 1.5 ng/ml on the trigger day did not negatively impact embryonic morphokinetic parameters, instead resulting in faster embryo development in the initial stages.

Behavioral, Physiological and Hormonal Changes in Primiparous and Multiparous Goats and Their Kids During Peripartum

The objective of this study was to evaluate the effect of maternal experience on behavior, progesterone and estradiol concentrations, weight and body condition in goats in the peripartum period, as well as the behavior and weight of the kids in the first hours after birth. Twenty multiparous and 10 primiparous dairy goats were enrolled. Behaviors such as walking, isolation, aggression, eating, inactivity and rumination were recorded in females during the last three days prior to the day of delivery, as well as the behaviors of the dam and her progeny in the first 2 h postpartum and the ability of the mother to discriminate between own kid and the alien one (selectivity test) at 2 h postpartum. Gestational and postpartum plasma progesterone and estradiol concentrations were determined. In the prepartum period there was a higher percentage of primiparous goats that were eating and remained less inactive compared to multiparous goats (p < 0.05). During the postpartum period, the lick latency to the kid was shorter in multiparous than in primiparous goats (p = 0.015). There was a greater proportion of kids born from multiparous (67%) than from primiparous (27%) that stood up in the first 30 min postpartum (p < 0.0001). Therefore, the proportion of multiparous kids (33%) than primiparous kids (9%) that were nursed within the first hour after birth was also higher (p < 0.0001). Progesterone and estradiol concentrations were not affected by maternal experience, neither in the prepartum nor the postpartum. In conclusion, maternal experience affected some behaviors at peripartum of the dam and vitality in the kid; however, the differences in behavior were not accompanied by differences in the variations in circulating estradiol and progesterone levels in the dam.

The effect of the menstrual cycle phases on back squat performance, jumping ability and psychological state in women according to their level of performance -a randomized three-arm crossover study

ObjectiveThe influence of the menstrual cycle on practical power performance such as barbell back squats and jumping performance in women has not yet been fully investigated. In addition, the performance level of athletes has not been considered in previous studies. This study aimed to investigate the influence of different cycle phases on acute back squat performance, jumping ability and psychological state concerning the performance level.Methods24 female strength athletes (age: 25.2 ± 3.3 years; height: 169.5 ± 3.4 cm; body weight: 67.7 ± 7.3 kg) were recruited for the study. Level of performance was classified according to Santos et al. (intermittent (n = 13), advanced (n = 6), highly advanced (n = 5)). Participants were tested for 1RM barbell back squat and jumping performance (countermovement and squat jump) as well as two questionnaires assessing their psychological states in the menses (M), late follicular phase (FP) and mid-luteal phase (LP) in three MC. Saliva estradiol and progesterone concentrations with a menstrual cycle diary were used to confirm a normal MC. A principal components analysis for power performance, well-being, relaxation and alertness was carried out and a linear mixed model was used for statistical evaluation.ResultsNo significant differences were found between the MC phases in performance scores (p > 0.05), readiness (p > 0.05) and alertness (p > 0.05). However, a high correlation between MC phases, performance level and back squat performance was detected. Correlation analyses indicate that a higher performance level results in a higher variation depending on the MC of the squat performance. For well-being, a significantly lower score was detected in M than in FP and LP.ConclusionIn general the performance score of the lower body is not influenced by the MC. If strength performance and jumping ability are considered separately, there are indications that strength capability is influenced at a higher performance level. In addition, individual variance was also observed, so this should also be considered. However, further studies are needed to confirm this assumption due to the small sample sizes of the individual performance levels.Trial registrationGerman registry for clinical studies (DRKS00034816, Date: 08/01/2024).

Evaluation of Four Different Automated Activity Monitoring Systems to Identify Anovulatory Cows in Early Lactation.

The objective of this study was to evaluate four different automated activity monitoring (AAM) systems to identify anovulatory cows in early lactation. A total of 852 lactating Holstein cows (221 primiparous and 631 multiparous cows) from four commercial dairy herds were enrolled. On each farm, cows were equipped with a respective AAM system (SB: Smartbow; HT: Heatime; DP: Delpro; and CM: CowManager). Each cow was sampled three times within the voluntary waiting period (VWP) in a two-week interval to detect the blood progesterone (P4) concentration. Cows were classified based on the concentration of P4 as follows: (1) none of the three blood P4 concentrations exceeded 1.0 ng/mL (anovulatory); (2) at least one of the three blood P4 concentrations was above 1.0 ng/mL (ovulatory). Cows were classified based on estrus alerts as follows: (1) no estrus alert was detected by an AAM system from 7 to 60 DIM (anestrus); (2) at least one estrus alert was detected by an AAM system from 7 to 60 DIM (estrus). Accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were calculated for each AAM system for anovulatory cows [(SB: 77.6%; 26.8%; 89.3%; 36.7%; and 84.1%); (HT: 79.2%; 63.6%; 83.6%; 52.8%; and 88.9%); (DP: 47.2%; 78.8%; 41.5%; 19.5%; and 91.6%); (CM: 80.5%; 23.7%; 92.7%; 39.1%; and 85.3%)].

The Impact of Gestational Diabetes Mellitus on Minipuberty in Girls.

Minipuberty is the second phase of physiological activation of the reproductive axis, playing a role in the postnatal development of sexual organs. The course of female minipuberty was found to be affected by low maternal vitamin D status and hypothyroidism during pregnancy. The aim of the current study was to assess the hormonal profile and the size of sexual organs in daughters of mothers with gestational diabetes mellitus. The study included three matched groups of infant girls: daughters of healthy women without metabolic disorders during pregnancy (group 1), daughters of women with poorly controlled gestational diabetes mellitus (group 2), and daughters of women with gestational diabetes mellitus adequately controlled during pregnancy (group 3). Urinary levels of gonadotropins, salivary levels of estradiol, testosterone, DHEA-S and progesterone, ovarian volume, uterine length and breast diameter were measured from postnatal month 1 to postnatal month 18. Concentrations of FSH, LH and estradiol were higher, while concentration of progesterone was lower in group 2 than in the remaining groups. There were no between-group differences in concentrations of testosterone and DHEA-S. Levels of LH, FSH, estradiol and progesterone correlated with maternal whole-blood levels of glycated hemoglobin. Group 2 was also characterized by the longest detection periods for LH and estradiol. Ovarian volume, uterine length and breast diameter were greater in group 1 than in the remaining two groups. Over the entire observation period, there were no differences in hormone levels and sizes of the sexual organs between groups 1 and 3. The obtained results suggest that poorly controlled, but not well controlled, gestational diabetes mellitus affects the course of female minipuberty.

A supergene affects androgen concentrations during early development in a bird with alternative reproductive morphs

Species with alternative reproductive tactics typically show pronounced phenotypic variation between and within sexes. In some species, this variation culminates in discrete reproductive morphs that are genetically determined, facilitating studies on how genetic variation translates into phenotypic variation. In ruffs (Calidris pugnax), an autosomal inversion polymorphism underlies three reproductive morphs (Independents, Satellites and Faeders), which differ in circulating steroid concentrations in adults. Yet, it remains unknown whether morph differences in steroid concentrations already arise before adulthood. We examined variation in circulating testosterone, androstenedione and progesterone concentrations between morphs and sexes in ruff chicks and juveniles and compared the differences to those in adults. Since measured hormone concentrations only provide momentary states and show high within- and between-individual variation, we took repeated measurements to compare means, variances and skewness between groups. We found clear differences between morphs but not the sexes in early life. Between morphs, androgen concentrations in young ruffs differed in variance and skewness, but not in their means. For testosterone, Independents had a higher variance than Satellites/Faeders, whereas for androstenedione, we observed the opposite pattern. For progesterone, we did not detect clear differences between groups. Skewness values mirrored differences in morph variances. Compared to adults, premature ruffs had lower androgen concentrations. In both life stages, we detected morph-specific associations between androgen concentrations: androstenedione concentrations increased with testosterone concentrations more in Satellites/Faeders than in Independents. These observed morph differences during early life are consistent with a supergene-mediated regulation of androgen variation that underlies the diversification of adult behavioural phenotypes.

Correlation between serum and saliva sex hormones in young female athletes.

Saliva sampling provides a practical noninvasive way for biological monitoring of steroid hormones but few studies have focused on saliva sex hormones in female athletes, and it is still unknown whether salivary concentrations are an accurate reflection of blood concentrations in this population. The aim of this study was therefore to evaluate the correlation between serum and saliva estradiol (E2) and progesterone (PG) in 10 young female athletes with normal menstrual cycle. Thirty blood and saliva samples were taken at rest in a fasting state in order to analyze E2 and PG concentrations, during 3 different hormonal periods: Status 1: low E2 and low PG (end luteal/early follicular); Status 2: high E2 and low PG (end follicular/peri-ovulatory); Status 3: high PG (mid-luteal). A strong correlation was found between serum and saliva PG (r=0.914, P<0.01), whereas a weaker but still significant relationship was observed between serum and saliva E2 (r=0.755, P<0.01). The most significant correlations obtained for E2 were during Status 2 (r=0.941, P<0.01) and during Status 3 for PG (r=0.817, P<0.01). Although it remains challenging to produce precise identification with only saliva samples for each menstrual cycle phase, results seem to show that saliva can provide a convenient substitute to serum for sex hormones in female athletes during longitudinal monitoring. This is of particular interest in this population, with the number of female athletes suffering from an altered menstrual cycle, sometimes even during regular menstruation.

The impact of enhanced early postnatal nutrition on the histomorphological features of the ovaries in fat-tailed Sanjabi ewe lambs.

This study aimed to investigate whether providing additional ad libitum feeding could improve reproductive characteristics in prepubertal ewe lambs. Twelve Sanjabi ewe lambs, approximately one-month-old and with an average initial weight of 11.6 ± 1.3kg, were monitored over a 3-month period, which was divided into three distinct periods of 30 days each. The first period spanned from the lambs' age of one to two months, the second period covered their age range of two to three months, and the third period encompassed their age range of three to four months. The lambs were randomly assigned to either a control group (C) or a treatment group (T). The control group received a basal diet consisting of hay and milk, whereas the treatment group received an enhanced diet comprising hay, milk, and concentrate. Live weight (LW) and average daily gain (ADG) were measured weekly, and blood samples were collected from the jugular vein at days 30, 60, and 90 of the experiment to determine the concentrations of progesterone (P4) and estrogen (E2). Following the 90-day experiment, the lambs were humanely euthanized in accordance with established protocols. Subsequently, both ovaries from each animal were removed, weighed and subjected to a comprehensive histological analysis. The ovaries underwent fixation, embedding, and sectioning procedures, followed by staining with hematoxylin and eosin (H&E) to facilitate microscopic examination. Ovary microscopic sections from both C and T groups were examined to assess follicles at various developmental stages, including primordial follicle number (PreFN) and diameter (PriFD), primary follicle number (PriFN) and diameter (PriFD), secondary follicle number (SFN) and diameter (SFD), antral follicle number (AFN) and diameter (AFD), and granulosa layer thickness of secondary (GLTSF) and antral follicles (GLTAF). The impact of diet and three experimental periods on ADG and LW exhibited significant variations. There was a significant increase (P < 0.05) in LW in the T group compared to the T group, with noticeable differences in LW fluctuations observed during distinct periods. However, ADG was more prominent in the initial and second observation periods than in the third. The impact of diet and experimental periods on P4 and E2 concentrations was found to be negligible. The T group showed significant improvements (P < 0.05) in PreFD, PriFN, and GLTAF compared to the C group. This study suggests that ad libitum concentrate feeding may promote reproductive characteristics in prepubertal ewe lambs. Further research is needed to explore its potential impact on the age of puberty.

Heterotropic Allosteric Modulation of CYP3A4 In Vitro by Progesterone: Evidence for Improvement in Prediction of Time Dependent Inhibition for Macrolides.

Predictions of drug-drug interactions resulting from time-dependent inhibition (TDI) of CYP3A4 have consistently overestimated or mis-predicted (i.e. false positives) the interaction that is observed in vivo. Recent findings demonstrated that the presence of the allosteric modulator progesterone (PGS) in the in vitro assay could alter the in vitro kinetics of CYP3A4 TDI with inhibitors that interact with the heme moiety, such as metabolic-intermediate complex (MIC) forming inhibitors. The impact of the presence of 100 µM PGS on the TDI of molecules in the class of macrolides typically associated with MIC formation was investigated. Presence of PGS resulted in varied responses across the inhibitors tested. The TDI signal was eliminated for five inhibitors, and unaltered in the case of one, fidaxomicin. The remaining molecules erythromycin, clarithromycin, and troleandomycin, were observed to have a decrease in both potency and maximum inactivation rate ranging from 1.7-fold to 6.7-fold. These changes in TDI kinetics led to a >90% decrease in inactivation efficiency. In order to determine in vitro conditions that could reproduce in vivo inhibition, varied concentrations of PGS were incubated with clarithromycin and erythromycin. Resulting in vitro TDI kinetics were incorporated into dynamic physiologically-based pharmacokinetic (PBPK) models to predict clinically observed interactions. The results suggested that a concentration of ~45 µM PGS would result in TDI kinetic values that could reproduce in vivo observations and could potentially improve predictions for CYP3A4 TDI. Significance Statement The impact of the allosteric heterotropic modulator progesterone on the CYP3A4 time-dependent inhibition kinetics was quantified for a set of metabolic-intermediate complex forming mechanism-based inhibitors. We identify the in vitro conditions that optimally predict time-dependent inhibition for in vivo drug-drug interactions through dynamic physiologically-based pharmacokinetic modeling. The optimized assay conditions improve in vitro to in vivo translation and prediction of time-dependent inhibition.

Impact of progesterone concentration on hCG trigger day on clinical outcomes with cleavage-stage embryo transfer in in vitro fertilization cycles with an antagonist protocol

Objective: To investigate the impact of the progesterone concentration on human chorionic gonadotropin (hCG) trigger day in fresh cycles versus thawed transfer cycles (the freeze-all strategy) with an antagonist protocol, and to compare the differences in clinical outcomes. Methods: This retrospective cohort study included a total of 2 165 cycles conducted at Henan Provincial People's Hospital with cleavage-stage embryo (at least one top-quality) transfer between January 2017 and December 2023, with serum progesterone levels on hCG trigger day all≤6.34 nmol/L (i.e. 2 ng/ml). Multivariate logsitic regression analysis and curve fitting were performed based on different serum progesterone levels on hCG trigger day [≤3.17 nmol/L (i.e. 1 ng/ml) or 1-2 ng/ml]. Results: Multivariate regression analysis, by using cycle type (either fresh or frozen-thawed cycle) as the exposure variable, showed that the clinical pregnancy rate (≤1 ng/ml: OR=0.93, 95%CI: 0.75-1.14; 1-2 ng/ml: OR=1.05, 95%CI: 0.58-1.87) and live birth rate (≤1 ng/ml: OR=0.90, 95%CI: 0.71-1.13; 1-2 ng/ml: OR=1.53, 95%CI: 0.79-3.00) had no statistically significant differences in group of progesterone concentration ≤1 ng/ml or in group of 1-2 ng/ml. Using serum progesterone levels on hCG trigger day as a continuous variable for curve fitting analysis, the clinical pregnancy rate in fresh or thawed cycles showed no significant changes with increasing progesterone levels. Conclusions: In the antagonist protocol with cleavage-stage embryo transfer (at least one top-quality), when the serum progesterone level on hCG day is ≤2 ng/ml, there are no significant differences in clinical outcomes between thawed cycles and fresh cycles, including clinical pregnancy rate and live birth rate. Transferred in fresh cycles or choosing the freeze-all strategy could be selected based on the actual situation of the patients.

Vaginal cytology during the estrous cycle and early pregnancy of ewes

Changes in vaginal epithelial cells and blood progesterone and estrogen concentrations were compared between estrous cycle and early pregnancy in multiparous Sanjabee ewes. Twenty non-pregnant ewes were synchronized with intravaginal insertion of a controlled internal drug release (CIDR) device which was in place for 7 days, intramuscular administration of a GnRH analog (Alarelin acetate; 12.5 µg) at the time of CIDR insertion (Day -7), a PGF2α analog (D-cloprostenol sodium; 125 µg) and 500 I.U. of human chorionic gonadotropin (hCG) at the time of CIDR removal (Day 0). At Day 0, all ewes were introduced to four fertile rams and observed for estrous signs. Ewes exhibiting estrous signs were divided into two groups: MATED (confirmed pregnant with transrectal ultrasound scanning at Day 35) and NOT-MATED (confirmed non-pregnant with transrectal ultrasound scanning at Day 35). Vaginal smears and blood samples were taken daily and every other day, respectively, from the beginning of estrus for 20 days for cytology and hormone assay. Based on the results, during days 0 and 1 of the estrous cycle, only the percentages of intermediate cells differed between the groups (P &lt; 0.05). During days 2 to 4, there was no difference in the cell populations between the groups (P ≥ 0.05). During days 5 to 16, the percentages of all cell types except the parabasal cells differed between the groups (P &lt; 0.05). During days 17 to 20, the percentages of all types of vaginal epithelial cells differed between the groups (P &lt; 0.05). Progesterone concentrations increased gradually from days 0 to 14 of the estrous cycle in both groups, however, they decreased significantly afterward in the NOT-MATED group. Estrogen concentrations changes showed an opposite pattern to that of progesterone in the study groups. Collectively, vaginal cytology can be used as a useful tool in assessing hormonal and physiological characteristics of the female reproductive system and thus provides a more accurate understanding of the physiology of estrous cycle and early pregnancy in ewes, which can be used to improve reproductive management.

Effect of a Bacillus subtilis-based direct-fed microbial, on milk yield, milk components, feed intake and plasma hormones and metabolites in lactating Holstein cows

To investigate the effects of a Bacillus-based direct-fed microbial on milk production and related factors, a study was conducted on Holstein cows (n = 28) starting at 90 ± 11 DIM. The cows were divided into two dietary groups: a control group (CON) that received a total mixed ration (TMR), and a Bacillus-fed group (DFM) that received the same TMR along with a Bacillus subtilis product containing two strains (747 and 1781) in equal amounts. The study lasted for 25 wk and included both primiparous and multiparous cows. The cows were housed in a free-stall barn and were provided with ad libitum TMR, which was fed twice a day. Their daily meals and dry matter intake (DMI) were recorded during wk 1–4 and wk 19–25 using electronic feeders. Milk samples were collected weekly during morning and evening milkings and analyzed for milk fat, protein, lactose, and milk urea nitrogen (MUN). Blood samples were also collected weekly and analyzed for plasma glucose, insulin, cholesterol, insulin-like growth factor 1 (IGF1), and progesterone concentrations. Additionally, rumen fluid samples were collected and evaluated for various species of bacteria. One of the key findings was that daily 4 % fat-corrected milk production (FCM) was influenced by the interaction between treatment (DFM vs. CON) and parity (primiparous vs. multiparous), with multiparous DFM cows producing 11 % more FCM compared to CON. The DFM cows also had a higher milk fat percentage (4.41 % vs. 4.02 %) and MUN concentrations were 1.0 mg/dL higher in the DFM group. In terms of blood metabolites and hormones, multiparous DFM cows had lower plasma cholesterol, glucose, and insulin concentrations compared to CON multiparous cows. However, there were no significant differences between the groups in terms of plasma IGF1 concentrations, insulin-to-glucose ratios, and luteal phase plasma progesterone concentrations. The analysis of rumen fluid samples revealed that the abundance of Ruminococcus albus and Fibrobacter succinogenes group I was greater in the DFM cows compared to CON cows. Furthermore, during the periods when feed intake was measured, the DFM cows had a 9 % reduction in feed intake and a 14 % improvement in feed efficiency (FCM per DMI) compared to CON cows. In conclusion, the two-strain Bacillus product used in this study (Certillus) showed potential as an effective direct-fed microbial. It was found to increase feed efficiency and milk production by altering the composition of ruminal microbiota and metabolism in the cows.

Progesterone increases metabolism via the pentose phosphate pathway in bovine uterine epithelial cells.

During early pregnancy, glucose is essential for the uterine epithelium and the developing embryo. In cows, progesterone increases the secretion of glucose into the uterine lumen. The uterine epithelium can convert glucose to fructose, but other fates of glucose in the uterine epithelium have been sparsely investigated. Therefore, our objective was to investigate how progesterone influences glucose metabolism in immortalized bovine uterine epithelial (BUTE) cells. BUTE cells were grown to 80% confluence and treated with vehicle (DMSO) or 10 µM progesterone for 24h. Cells were collected and analyzed. Immunohistochemistry was performed on endometrial samples collected from the bovine endometrium on days 1 and 11 of the reproductive cycle. Progesterone treatment increased glucose consumption of BUTE cells. RNAseq identified 3,072 genes regulated by progesterone. KEGG analysis indicated that progesterone altered genes associated with metabolic pathways and glutathione metabolism. Manually examining genes unique to specific glucose metabolic pathways identified an increase in the rate-limiting enzyme in the pentose phosphate pathway-glucose-6-phosphate dehydrogenase. Functionally, a major product of the pentose phosphate pathway is NADPH, and progesterone treatment increased NADPH levels in BUTE cells. In cows, immunohistochemistry confirmed that glucose-6-phosphate dehydrogenase levels were higher in the uterine epithelium in the luteal phase when progesterone concentrations are high. Progesterone increased glucose-6-phosphate dehydrogenase expression and metabolism via the pentose phosphate pathway in the bovine uterine epithelium. This metabolism could provide substrates for cell proliferation, molecules to be secreted into the uterine lumen, or maintain reduction/oxidation balance in the uterine epithelium.

Regulator of G-Protein Signaling 2 Knockout in CD4+ T Cells Promotes Anti-Inflammatory T Cells, Enhancing Ovulation, and Oocyte Yield.

To determine the downstream effects on ovarian function and immune cell differentiation in the ovary and uterus using a model in which RGS2 was knocked out specifically in CD4+ T cells. Laboratory based experiments with female mice. Female congenic (fully backcrossed) and non-congenic (mixed strain) mice with CD4 T cell-specific RGS2 knockout. Four-week-old female CD4 RGS2 knockout (CD4 RGS2 KO ) mice and their littermate controls (CD4 RGS2 CTL ) were subjected to superovulation using pregnant mare serum gonadotropins. Oocyte numbers, lymphocyte populations in the ovary and uterus, and serum estradiol and progesterone concentrations. In non-congenic (mixed strain) mice, CD4 RGS2 knockout (KO) promoted higher oocyte ovulation and increased uterine total leukocyte numbers. Similarly, congenic (fully backcrossed strain) mice showed higher oocyte numbers and increased uterine total leukocytes in the CD4 RGS2 KO mice compared to CD4 RGS2 CTL mice. Pro-inflammatory CD4+ T helper (T H ) 1 and T H 17 cell frequencies in the ovary and uterus were unchanged, while Treg and T H 2 cell frequencies were elevated, along with increased concentrations of estradiol and progesterone in the serum of CD4 RGS2 KO mice. Our study highlights the important role of RGS2 in CD4+ T cells within the context of reproduction. The dysregulation of immune responses due to RGS2 knockout in CD4+ T cells appears to enhance oocyte production. Further research is warranted to elucidate the precise mechanisms by which RGS2 influences reproductive outcomes, including its impact on fecundability, endometrial receptivity, and successful implantation.

The metabolomic composition of the oviductal fluid is controlled by the periovulatory hormonal context in Bos indicus cows.

In cattle, oviductal function is controlled by the ovarian sex-steroids estradiol and progesterone. Here, we tested the hypothesis that the exposure to contrasting sex-steroid milieus differentially impacts the oviductal fluid composition. Estrous cycles of non-lactating, multiparous Nelore cows were pre-synchronized and then synchronized with a protocol designed two induce ovulation of large (LF group) or small (SF group) follciles. Larger preovulatory follicle (day 0) and corpora lutea (day 4) and greater estradiol (day 0) and progesterone (day 4) concentrations were observed in the LF group. Four days after induced ovulation, oviductal fluid was collected post-mortem. Quantitative mass spectrometry was used to determine the concentration of amino acids, biogenic amines, acylcarnitines, phosphatidylcholines, lysophosphatidylcholines, sphingomyelins, hexoses, prostaglandins and related compounds. Multivariate analyses (OPLS-DA) were conducted to compare the metabolomic signatures of oviductal fluids. Correlation network analysis was conducted to measure the strength and hierarchy of associations among metabolites. Of the 205 metabolites quantified, 171 were detected in at least 50% of the samples and were included in further data analysis. After OPLS-DA analysis, samples of the LF and SF were divided clearly into two non-overlapping clusters. Twenty metabolites had different or tended to have different concentrations in the oviductal fluid when comparing groups. Seven of these 20 analytes had greater concentrations in LF cows. Moreover, total sum of biogenic amines, phosphatidylcholines, and prostaglandins were higher in the SF group. The correlation network showed that the LF group metabolites' concentrations were highly intercorrelated, which was not observed in the SF group. We concluded that the periovulatory endocrine milieu regulates the composition of the oviductal fluid.

Progesterone Concentrations during Uterine Involution in Sahelian Goats

Progesterone Concentrations during Uterine Involution in Sahelian Goats