P4 Levels Research Articles

EFFECTS OF INJECTABLE MINERAL AND VITAMIN SUPPLEMENTATION ON THE FERTILITY OF DAIRY COWS IN FTAI PROTOCOL

The study aimed to assess the impact of injectable mineral and vitamin supplementation, specifically Kit Adaptador®MIN and Adaptador®VIT (Biogénesis Bagó, Brazil), administered at the initiation of a fixed-time artificial insemination (FTAI) protocol, on the reproductive performance and fertility of Holstein cows. One hundred animals were divided into a non-supplemented control group (CG, n=50) and a treated group (TG, n=50). The TG received mineral and vitamin supplementation (1 mL/100kg live weight, IM) on the first day of the FTAI protocol (D0), along with intravaginal progesterone (P4) implantation and estradiol benzoate administration. Blood samples were collected at D0 and D17 to assess superoxide dismutase (SOD) enzyme activity and P4 levels. Ovarian and uterine ultrasound evaluations were conducted at D0 and D17, with follow-up exams performed 30 days post-FTAI for pregnancy diagnosis. Results showed a larger corpus luteum diameter in supplemented cows (CG=2.14±0.11 cm vs TG=2.64±0.11 cm, P=0.0037), along with increased SOD activity (CG=22% vs TG=25.36%, P=0.045). Moreover, there was a significant difference in pregnancy rates between groups (CG=18.18% vs TG=39.13%, P=0.0029). Injectable mineral and vitamin supplementation at the start of the FTAI protocol may impact post-ovulation corpus luteum development, enhance pregnancy rates, and stimulate essential endogenous antioxidant mechanisms in Holstein cows.

Effects of cloprostenol and oxytocin on ultrasonographic changes of uterine horns in postpartum primiparous Hu sheep

This study aimed to evaluate the effects of exogenous hormones on postpartum ultrasonographic changes of uterine horn in Hu sheep. 90 nulliparous Hu sheep were divided into three groups according to the number of lambs. Each group was further randomly subdivided into three groups (n = 10), a control group (sodium chloride), a PG group (cloprostol), and a PG+OT group (cloprostol + oxytocin). The effects of different treatments on ultrasonographic changes of uterine horn and reproductive performance of ewes were determined after the treatment. Then, 90 nulliparous ewes were divided into a control group, PG group, and PG+OT group according to hormone injections received. Each group (n = 30) consisted of 10 single, 10 twin, and 10 triple ewes. Changes in the levels of hormones P4, LH, FSH, and E2 in the serum of postpartum ewes, the time of the first estrus, and the conception rate were detected. Results showed that the higher the lambing rate, the slower the rate of ultrasonographic changes of uterine horn. The rate of ultrasonographic changes of uterine horn in the PG+OT group was faster than that in respective PG and the control groups. Caruncle diameter was not affected by the number of lambs or hormone treatment. After delivery, the concentrations of P4 and LH increased rapidly and remained at a relatively high level, while the concentration of E2 increased in all ewes. The PG+OT group had lower P4 concentrations compared to the control group, but higher LH, FSH, and E2 levels. The time to first postpartum estrus was significantly shorter in the PG+OT group, but differences in pregnancy rate were not significant. Thus, PG+OT treatment can significantly accelerate postpartum ultrasonographic changes of uterine horn, which can prepare postpartum ewes for the next reproductive cycle.

Correlation Between Luteal Phase Serum Progesterone Levels and Pregnancy Outcome in Frozen Embryo Transfer Cycles: A Prospective Cohort Study

Objectives To study the correlation between luteal phase serum progesterone (P4) levels and pregnancy outcome in frozen embryo transfer (FET) cycles and to find out the cut-off level of luteal phase serum P4 which favours successful pregnancy outcome in FET cycles. Material and Methods This prospective cohort study included 100 women undergoing hormone replacement therapy (HRT)-FET cycle at Akanksha IVF Centre from December 2023 to June 2024, fulfilling the inclusion and exclusion criteria. Serum P4 levels were measured for all the patients in the luteal phase (days 21–23) after FET with intramuscular P4 as luteal phase support. Pregnancy outcome was assessed in terms of implantation rate, biochemical pregnancy rate, clinical pregnancy rate (CPR), first-trimester miscarriage rate and ongoing pregnancy rate. Results The most favourable pregnancy outcomes were observed at serum P4 levels 25.1-35 ng/mL. A statistically significant association was seen between the luteal phase serum P4 levels and implantation rate (36.08%), biochemical pregnancy rate (4%), CPR (52%) and ongoing pregnancy rate (42%). According to the receiver operating characteristic (ROC) curve analysis, the optimal cut-off value for favourable outcomes was determined to be 22.3 ng/mL. However, the analysis also indicated that the luteal phase serum P4 levels did not reliably predict clinical or ongoing pregnancy. Conclusion Larger studies are needed to establish a threshold level of serum P4 in the luteal phase that can differentiate between successful and unsuccessful implantation. However, it is still uncertain whether the issue of unsuccessful implantation can be resolved once it is detected on days 21–23 of the HRT-FET cycle.

Exogenous progesterone rescue in patients with low mid-luteal serum progesterone levels undergoing true natural vitrified-warmed blastocyst transfer.

To explore whether a 25mg subcutaneous progesterone daily rescue daily improves the reproductive outcomes in patients with low serum progesterone (P4) levels (7-10ng/mL), measured one day before true natural cycle (t-NC) frozen embryo transfer (FET). A cohort study of 192 women undergoing t-NC warmed blastocyst transfer. Patients were stratified into three different groups based on serum P4 levels on the FET-1day: patients who had serum P4 levels of 7-10ng/mL and underwent rescue progesterone administration (rescue group), patients with serum P4 levels of 7-10ng/mL without progesterone administration (non-rescue group), and patients with serum P4 > 10ng/mL on FET-1day (control group). The primary outcome was possible differences in live birth rate (LBR) between groups. The LBRs for the serum P4 7-10ng/mL without rescue, 7-10ng/mL with rescue, and > 10ng/mL (control) groups were 41%, 46%, and 52%, respectively (p = 0.61). The estimated adjusted probability of live birth for serum P4 7-10ng/mL without rescue, 7-10ng/mL with rescue, and > 10ng/mL (control) groups were also comparable: 43.5% (95% CI, 20.0-70.4%), 49.8% (95% CI, 28.1-71.6%), and 57.4% (95% CI, 44.0-69.8%), respectively. Serum P4 levels higher than 7ng/mL seem to secure LBRs in patients undergoing t-NC FET. A rescue policy consisting of a daily subcutaneous 25mg progesterone dose in patients with serum P4 levels 7-10ng/mL does not further enhance LBRs when compared to those patients with similar serum P4 levels without rescue.

Diagnosis and hormonal treatment of cystic ovaries associated with uterine disorders in Egyptian buffaloes: ultrasonography, histopathological and serological investigations

Cystic ovarian disease (COD) with uterine abnormalities is a postpartum reproductive pathology in Egyptian buffaloes causing significant economic losses. In this study, we aimed to employ various diagnostic methods for detecting cystic ovarian disease (COD) in Egyptian buffaloes. tour study assessed the effectiveness of the GnRH/PGF2α protocol as a treatment strategy. Our goal was to determine if this protocol could effectively reduce economic losses associated with cystic ovarian disease and improve herd fertility in Egyptian buffaloes. Eighty Egyptian buffalo cows were included in this study. They were identified to have follicular cysts through rectal examination, which was confirmed by ultrasonography. These buffaloes were then divided into two main groups: the COD Control (untreated) (GA) (n = 40) and COD group (GB) (n = 40) treated with GnRH/PGF2α. According to our immunological studies, buffaloes in the COD-treated group (GB) exhibited significantly lower serum levels of pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) compared to the control group. This observation was consistent with the decline in E2 levels and the increase in P4 levels (p < 0.01–0.001) observed in the treated animals compared to the untreated group. Furthermore, serum cortisol and glucose concentrations decreased in COD-treated buffaloes. Histopathological examination of ovaries and uterine tissue from slaughtered COD buffaloes has revealed significant structural alterations. These include the presence of ovarian cysts of varying sizes with vacuolar degeneration. Additionally, lymphoplasmacytic endometritis was observed in the uterine tissue of affected animals, featuring degeneration and desquamation of the endometrial lining accompanied by infiltration of mononuclear inflammatory cells. Severe and prolonged cases of COD, which did not respond to treatment, exhibited marked adverse pathological changes upon histopathological assessment of the genital tract. In conclusion, hormonal treatment with GnRH/PGF2α appears to be effective in treating COD-affected animals. The study provides valuable insights into the immunological, biochemical, and histopathological aspects of cystic ovaries associated with uterine disorders in Egyptian buffaloes, while also evaluating hormonal treatment for cystic ovarian disease as a means to minimize economic losses and improve herd fertility in this species.

Dan’e fukang decoction reduces hemorrhage in a rat model of mifepristone induced incomplete abortion and may correlate with cell adhesion molecule signaling interference

Ethnopharmacological relevanceDan’e fukang decoction (DFD) is a traditional Chinese medicine formula. DFD obtains 10 herbs, including Salvia yunnanensis C.H.Wright (Zidanshen), Curcuma zedoaria (Christm.) Roscoe (Ezhu), Angelica sinensis (Oliv.) Diels (Danggui), Cyperus rotundus L. (Xiangfuzi), Corydalis yanhusuo (Y.H.Chou & Chun C. Hsu) W.T.Wang ex Z.Y.Su & C.Y.Wu, Bupleurum marginatum Wall. ex DC. (Yanhusuo), Sparganium stoloniferum (Buch.-Ham. ex Graebn.) Buch.-Ham. ex Juz. (Sanleng), Panax notoginseng (Burkill) F.H.Chen (Sanqi), Paeonia lactiflora Pall. (Shaoyao) and Glycyrrhiza uralensis Fisch. (Gancao). DFD is now clinically used for the treatment of menstrual irregularities, dysmenorrhea and menstrual discomfort caused by blood stasis and easing of endometriosis. Based on this, it is reasonable to presume that DFD may be effective in treating incomplete abortion and reducing postpartum bleeding, but no specific studies have been reported so far. Aim of the studyTo investigate the efficacy of Dan’e fukang decoction (DFD) in reducing prolonged vaginal bleeding followed by mifepristone induced incomplete abortion and explore the mechanisms of action of DFD in treating incomplete abortion. MethodsAn incomplete abortion model of rat was established by single intragastrically administered 8.5 mg/kg mifepristone on the 7th day of pregnancy. From the 8th day of pregnancy, the abortive rats were administered solvent, a positive control drug, or different doses of DFD, respectively for seven consecutive days. The efficacy of DFD was assessed by measuring the vaginal bleeding volume of the rats. Four coagulation parameters and platelet counts were measured. Hematoxylin and eosin (HE) staining was performed to evaluate pathological changes in the uterine embryos. Serum levels of progesterone and estrogen were measured using ELISA. Network pharmacology and transcriptomics were used to predict potential targets and pathways for DFD to reduce hemorrhage. The levels of mRNA related to cell adhesion molecules (CAMs) were detected by RT-qPCR. The levels of progesterone and estrogen receptors and the proteins associated with CAMs pathway in uterine tissues were detected by Western Blot. ResultsDFD significantly reduced the volume of vaginal bleeding of the abortive rats and significantly downgraded the pathological scores of uterine embryos. DFD significantly increased serum levels of E2, and had no impact on serum levels of P4 and the protein expression of ER and PR in the uteri of the abortive rats. Pathways in cancer, lipid, focal adhesion and immune-related signaling were predicted to be influenced by DFD via the analysis of network pharmacology. The CAMs signaling was found the most critical pathway regulated by both mifepristone and DFD via RNA-seq assay, followed by axon guidance, basal cell carcinoma, hippo signaling pathway and neuroactive ligand-receptor interaction. Combining the two analytical methods, ICAM-1 was predicted likely the key targeted gene by DFD. Finally, DFD was validated to decrease the protein expression of ICAM-1, ITGB2, ITGB7 and RASSF5 in the uterine tissues, which correlated to suppress the CAMs signaling pathway. ConclusionDFD significantly reduced hemorrhage. DFD significantly increased the serum levels of E2 and inhibited CAMs signaling pathway, which was likely to be involved in the mechanism of action of DFD facilitating residual uterine embryo expulsion in the rat model of incomplete abortion.

Effects of exogenous progesterone on the ovine serum progesterone profile during the non-breeding season

Exogenous progesterone (PRG) sources are used to induce ovine estrus during the non-breeding season. Intramuscular injection (IM) of PRG is an uncommon route for this purpose. This study investigated serum progesterone (P4) levels after IM administration of 50 mg of two different PRG products. Ten ewes were selected and received two consecutive PGF2α injections at 24-h intervals. Serum samples were collected (Time 0), followed by the injection of single doses of Fertigest® or Vetagesterone®. Serum P4 levels were assayed at times (T) 0, 30 min, 3,6,12,24,48,72,96, and 120 h after injection. A large variation in the serum P4 levels at T0 was detected. The serum P4 level at T0 was 4.3 ng/mL, which is considerable for anestrus ewes, following PGF2α administration. The mean time to reach maximum serum P4 concentration was 5.8 h, with a mean concentration of 22.5 ng/mL. Serum P4 levels returned to basal values (T0) after 48 h. An increased P4 production rate (PR) was detected following PRG administration, which continued for 24 h. A single injection of PRG reduced the variety of serum P4 levels from 24 h onward. The two different PRG generated different serum P4 concentrations. This study indicates an unusual change in serum P4 levels in anestrus ewes following PGF2α administration. 48 h after 50 mg PRG injection, the plasma levels of P4 reached basal levels in anestrus ewes.

Higher serum progesterone level has no negative impact on live birth rate in frozen embryo transfer

Optimal synchronisation between the endometrium and the embryo is key to the success of frozen embryo transfers (FET). It is achieved by administering different doses of exogenous oestrogen and progesterone (P4). The negative impact of low levels of P4 on FET has been reported, but there is a lack of knowledge about which levels are most appropriate. We aimed to evaluate whether high serum P4 levels measured on FET day affect the probability of having a baby at home. This retrospective cohort study includes 7546 FET cycles performed with hormone replacement therapy (HRT) for artificial endometrial preparation. Patients were divided into three groups according to P level on FET day (ng/mL): P4 < 20(n = 6623), P4 ≥ 20–40 (n = 770) and P > 40 (n = 146). Female age was per group P4 < 20 = 38.7 ± 3.1, P ≥ 20–40 = 39.0 ± 3.0, and P4 > 40 = 38.1 ± 2.9 years old (p = 0.53). The group with the highest progesterone levels found lower ongoing pregnancy rates without statistical significance (56.3 % (P4 < 20) vs 56.8 % (P4 ≥ 20-<40) vs 51.4 % (P4 > 40); p = 0.5). Miscarriage rate was also higher, although not significantly: 16.2 % (P4 < 20) vs 15.0 % (P4 ≥ 20-<40) vs 18.0 % (P4 > 40) (p = 0.6). These findings were explored with an adjusted analysis. A higher, but not significant, odds of miscarriage was observed when P4 > 40 ng/ml, aOR = 1.14 (0.75–1.74) (p = 0.55) whereas no such association was found with P4 ≥ 20–40 ng/ml (aOR = 0.92 (0.74–1.15) (p = 0.46)). The probability of livebirth is similar when the patient was P4 > 40 (aOR = 0.77 (0.51–1.15) (p = 0.20)) than when P4 ≥ 20-<40 (aOR = 0.94 (0.79–1.11) (p = 0.47). In conclusion, women with elevated serum P4 levels on the day of FET after HRT do not find their probability of live birth impaired.

Vascular Alterations in Uterine and Ovarian Hemodynamics and Hormonal Analysis throughout Pregnancy Loss in Cows under Heat Stress.

This current study examined the ovarian (OA) and middle uterine arteries (MUA) blood flow under heat stress conditions regarding hormonal status (progesterone (P4), estradiol (E2), and nitric oxide metabolites (NOMs) assays). Eighteen pluriparous cows were examined, with twelve only being subjected to the natural mating as the other six animals were not bred. Pregnancy diagnosis was confirmed at day 30 by embryonic heartbeat and CL graviditatis (n = 6; pregnant), but some animals (n = 6) showed early embryonic death (EED), with the presence of control cows (n = 6). In the pregnant group, luteal diameter (cm) increased after mating, while in the pregnancy loss group it increased (p < 0.05) until reaching day 38 (1.41 ± 0.03), then decreased again. Luteal vascularity was declined in cows with EED after day 36 (p < 0.05) and reached its lowest level at day 50. P4 levels elevated in cows with EED until day 36 (13.64 ± 0.11) then declined. Both ipsilateral OA and MUA Doppler indices were declined in both groups except in cows who suffered from EED; both were elevated from day 38 until day 50 after mating. Ipsilateral peak systolic point (PSV cm/sec) of OA and MUA was elevated in both groups (p < 0.05), but in cows with EED this parameter was declined. E2 and NOMs levels were declined in cows with EED from day 40 and day 38 after mating (p < 0.05), respectively. This study provided novel data on the relations among the luteal diameter, E2, P4, and NOM levels, and luteal hemodynamics that predicts the amount of blood supply, which acts as a sensitive parameter to detect the alterations in luteal function during the first 50 days after mating.

Differences in perinatal complications and serum hormone levels due to uterine endometrial preparation methods in frozen-thawed embryo transfer.

In frozen-thawed embryo transfer (FET), differences in endometrial preparation methods affect the incidence of perinatal complications. However, the underlying causes are unclear. We aimed to investigate whether serum E2, P4 levels are associated with perinatal complications. This is a retrospective cohort study, involving 306 successful FET pregnancies from 2017 to 2022. Participants were divided into Natural Cycle (NC) and Hormone Replacement Cycle (HRC) group. We compared serum hormone levels, maternal backgrounds, and perinatal outcomes and complications. Furthermore, within the HRC group, serum hormone levels were compared for perinatal complications previously reported to show differences in incidence rates depending on the method of endometrial preparation. HRC exhibited significantly higher serum E2 levels during the implantation period, but lower P4 levels during ovulation, implantation, and pregnancy test period compared with NC. HRC also had significantly higher rates of postpartum hemorrhage (PPH) and placenta accreta spectrum (PAS). There was no association found between perinatal complications more likely to occur in HRC and serum E2, P4 levels. In HRC, there were more occurrences of PPH and PAS. Although serum E2, P4 levels during FET did not correlate with perinatal complications.

Serum Progesterone Level on the Day of Embryo Transfer and Clinical Pregnancy Rate in Patients Undergoing Down-Regulated Frozen Embryo Transfer Cycles; Prospective Observational Study

Abstract Background Serum Progesterone concentration &amp;lt;5 ng/ml on the day of frozen embryo transfer (FET) is associated with significantly decreased pregnancy and live birth rates. Aim of the Work This study aims at finding the critical cutoff value of serum progesterone on day of FET that significantly affects pregnancy rates. Patients and Methods The current study was conducted at Ain Shams University Maternity Hospital- IVF Unit during the period between April 2022 and December 2022. It included a total number of 108 patients who either were assessed for eligibility of the study, of which, 14 were excluded due to non-matching inclusion criteria or had exclusion criteria. 4 patients did not consent to participate in the study and 86 patients matched. They were preaped for frozen emberyo transfer according to IVF unit protocol at Ain Shams university and serum progesterone level on day of emberyo transfer was assessed and matched to clinical pregnancy rate. Results Progesterone on day of embryo transfer was significantly lower in cases with positive chemical pregnancy with a cut off value ≤39.2 ng/mL. Conclusion Progesterone is essential for establishing and maintaining embryo implantation and pregnancy; also, P4 is used for luteal phase deficiency in infertility treatment. Yet, progesterone therapy’s optimal timing and dose can influence the impact of P4 on pregnancy. Also, low or elevated P4 level can negatively impact embryo transfer timing and pregnancy rate. This study demonstrated that age, BMI and progesterone on day of embryo transfer had significant low diagnostic performance in predicting positive chemical pregnancy This study illustrated that no statistical significant difference according to chemical pregnancy regarding hormonal profile Before the start of estradiol valerate. Progesterone on day of embryo transfer was significantly lower in cases with positive chemical pregnancy.

Comparative analysis of reproductive hormones, serum biochemical indexes and ovarian metabolites in Muscovy breeder duck at different laying stages

The hypothalamic-pituitary-gonadal (HPG) axis regulates egg laying through control hormones secretion in poultry. In this study, the serum hormones (12 samples per stage), serum biochemical indexes (12 samples per stage), and ovarian metabolites (8 samples per stage) of Muscovy breeder ducks were detected at prelaying stage (PT), start of laying stage (ST), high laying stage (HT), and the end of laying stage (ET). The serum hormones of Muscovy ducks were measured at 8:00, 13:00, 18:00, 23:00, and 4:00 within 1 d. The TG, TP, ALB, and GLB were significantly increased, while HDL-C was significantly decreased at ST as compared to PT (P < 0.05). Serum Na, Cl, Ca, P, and K showed significant rise at ST as compared to PT. Serum Na, Cl, Ca, and K were significantly declined, while P was significantly increased at ET as compared to HT (P < 0.05). Serum FSH, LH, PRL, E2, P4 levels peaked at ST (P < 0.05) with only FSH and LH fluctuated significantly within 1 ovulation cycle at ST (P < 0.05). Differential metabolites showed continued ovarian aging. The decline of nucleic acid metabolism occured in ST, the decline of sugar metabolism occurred in ET, and the decline of amino acid metabolism continued at all stages. Temporal expression patterns and correlation analyses indicated a high correlation between ovarian cAMP and serum reproductive hormone levels across different reproductive stages. In conclusion, this study revealed the changes in serum hormones, serum biochemical indicators, and ovarian metabolites, as well as the relationship between serum hormones and ovarian metabolites.

Effects of progesterone administered to maternal rats during the teratogenesis-sensitive period through different routes on neurobehavior and gut microbiota in offspring

Objective: Progesterone (P4) is the optimal agent for luteal support in assisted reproductive technologies. Despite the availability of various formulations, the impacts of P4 treatment administered through different routes on offspring growth remain unevaluated. This study aimed to investigate and compare the effects of P4 administration through three different routes in pregnant rats during the teratogenesis-sensitive period on fertility outcomes and the behavior, plasma P4 and pregnenolone levels, and alterations in gut microbiota in the filial generation (F1) of offspring. Methods: Female rats that mated successfully were randomly classified into four groups: Group 1, the normal control group; Group 2, the IG group, in which utrogestan was administered via intragastric gavage; Group 3, the IM group, in which P4 was administered via intramuscular injection; and Group 4, the VAGIN group, in which P4 sustained-release vaginal gel was administered through the vagina. Except for the control group, P4 was administered in other groups through their respective routes for 13 consecutive days daily during the teratogenic-sensitive gestation period (days 6–18). The fertility outcomes of the maternal rats were observed, and open-field and three-chamber social preference tests were conducted to assess anxiety and social behavior, along with an exhaustive swimming experiment to evaluate the motor endurance of the offspring. Plasma P4 and pregnenolone levels in the offspring were measured, and fecal samples were assessed using gut microbiota sequencing and subsequent bioinformatics analysis. Results: No significant alterations in fertility outcomes were observed through three P4 administration routes—intragastric gavage, intramuscular injection, and vaginal administration—in rats. However, in behavioral experiments in offspring, a significant increase was observed in swimming time in the IG group than in the control group (P = 0.013). Furthermore, the offspring in the IG group exhibited a marked decrease in non-social behaviors compared with the control group (P = 0.030). No significant behavioral changes were observed in the F1 offspring in the open-field test. Additionally, the offspring in the IG group exhibited a significant elevation in plasma pregnenolone levels compared with the control group (P = 0.019) 30 days after birth. No notable differences in the α-diversity of gut microbiota were observed among the groups. However, significant differences in β-diversity were observed in both the IG and IM groups compared with the control group, indicating that P4 treatment via the two routes exhibited a significant impact on the gut microbiota composition of F1 offspring. Distinct differences were observed in the gut microbiota of the offspring among the three P4 treatment groups, with 22, 28, and 33 differential bacterial taxa identified in the IG, IM, and VAGIN groups, respectively, using linear discriminant effect size analysis, whereas Olivella appeared exclusively in the control group rather than in the P4 treatment groups. Conclusion: Oral administration of P4 to maternal rats during the teratogenesis-sensitive period enhanced motor endurance and plasma pregnenolone levels and altered the composition of the gut microbiota in the offspring. Our findings demonstrated different effects on offspring growth when P4 is administered orally, intramuscularly, or vaginally.

Electro-responsive Molecularly Imprinted Polymers (E-MIPs) for Rapid Detection of Progesterone in Human Serum Samples.

Rapid and reagent-free detection of progesterone (P4) is crucial in point-of-care (POC) measurement due to its important role in the human endocrine and central nervous system. Currently available technologies for P4 detection are often not rapid or require reagents, limiting their use in POC settings. In this work, a self-signaling electrochemical sensing platform for rapid detection of P4 is developed by using electroactive molecularly imprinted polymers (E-MIPs). E-MIPs possess the ability to transduce the molecular recognition event into a measurable electrochemical signal. The E-MIPs are prepared by bulk copolymerization of acrylic acid (AA) and ferrocenylmethyl methacrylate (FcMA) in the presence of P4 followed by its removal from the polymer matrix. By incorporation of ferrocene moieties, the MIP gains intrinsic electroactivity, enabling label-free and direct electrochemical detection of P4 levels. Electrochemical techniques, including cyclic voltammetry and electrochemical impedance spectroscopy, were employed to elucidate the electron transport reaction responsible for the MIP's intrinsic electroactivity. Additionally, the predetermined complementary binding sites confined to the small working area of the screen-printed carbon electrode (SPCE) within the E-MIP matrix facilitate rapid P4 binding, achieving completion within 120 s. The developed E-MIP sensing platform was used for the rapid detection of P4 levels in human serum samples. The short incubation times reported for the E-MIP sensing platform would be beneficial in minimizing the nonspecific binding in the real-world samples. We believe that the proposed E-MIP sensing platform can be applied to clinical samples for detecting fluctuations in P4 levels.

SERUM DYNAMICS OF SOME REPRODUCTIVE HORMONES IN POSTPARTUM ANOESTRUS FRIESIAN X BUNAJI COWS TREATED WITH TWO DOSAGE REGIMENS OF OVSYNCH PROTOCOL

Fertility of the Post-partum period in dairy cattle is negatively influenced by the incidence of anoestrus. This condition is characterized by the absence of eostrus behaviour which may be due tophysiologic or pathologic factors. This study therefore, was aimed to compare the serum profiles of follicle stimulating hormorne (FSH), luteinising hormone (LH) and progesterone (P4) in response to the administration of two doses of gonadotrophin releasing hormone (GnRH, Lecirelin®) in combination with prostaglandin F2 alpha (PGF2α, Synchromate®) ovsynch protocol in apparently healthy, post-partum anestrous Friesian x Bunaji cows. Fifteen (n=15) animals were randomly divided into three groups and treated as follows: Group I (n=5) animals (control); Group II (n=5) animals received ovsynch 50 µg GnRH treatment; Group III (n=5) animals received 100 µg GnRH ovsynch treatment. Blood samples were collected at 15 to 30 minutes intervals for 6 hours to determine serum concentration of FSH and LH and once daily for 12 days after the injection of the first GnRH to determinate the concentration of plasma P4 using Enzyme linked immunosorbent assay kits. Peak values of FSH and LH were significantly (p&lt;0.05) different on days 0 and 9.The plasma P4 levels increased among treated cows following injection of GnRH reaching a peak on day 7 compared to the control group. It was therefore concluded that the ovsynch protocol at the dosages indicated enhanced FSH, LH and P4 profiles compared to control cows and could initiate the onset of ovarian activity in the post-partum anoestrus Friesian x Bunaji cows.

Unraveling the impact of hyperleptinemia on female reproduction: insights from transgenic pig model

BackgroundInfertility is a growing global health concern affecting millions of couples worldwide. Among several factors, an extreme body weight adversely affects reproductive functions. Leptin is a well-known adipokine that serves as an endocrine signal between adiposity and fertility. However, the exact mechanisms underlying the effects of high leptin level on female reproduction remain unclear.MethodsTransgenic pigs overexpressing leptin (♀) were produced by backcrossing and screened for leptin overexpression. The growth curve, fat deposition, reproductive performance, apoptosis, serum hormones and cholesterol production, RNA sequencing, and single-nucleus RNA sequencing (snRNA-seq) of the leptin-overexpressing pigs and wild-type group were evaluated.ResultsTransgenic pigs overexpressing leptin (♀) were obtained, which exhibited significantly reduced body weight, body size, and back fat thickness. These pigs manifested a late onset of puberty (330 ± 54.3 vs. 155 ± 14.7 days), irregular estrous behavior characterized by increased inter-estrous interval (29.2 ± 0 vs. 21.3 ± 0.7 days), and more number of matings until pregnancy (at least 3 times). This reproductive impairment in leptin pigs was related to hormonal imbalances characterized by increased levels of FSH, LH, prolactin, E2, P4, and TSH, altered steroidogenesis such as increased levels of serum cholesterol esters along with steroidogenic markers (StAR, CYP19A), and ovarian dysfunctions manifested by neutrophilic infiltration and low expression of caspase-3 positive cells in the ovaries. Moreover, bulk RNA sequencing of the ovaries also revealed neutrophilic infiltration followed by upregulation of inflammation-related genes. Furthermore, snRNA-seq reflected that leptin overexpression triggered immune response, suppressed follicle development and luteinization, resulting in metabolic dysfunction and hormone imbalance in the ovary.ConclusionsLow body weight in leptin overexpressing pigs adversely affects the reproductive performance, causing delayed puberty, irregular estrous cycles, and reduced breeding efficiency. This is linked to metabolic imbalances, an increased immune response, and altered ovarian functions. This study provides a theoretical basis for the complex mechanisms underlying leptin, and infertility by employing leptin-overexpressing female pigs.

Association between serum hormone levels in early pregnancy and risk of hypertensive diseases of pregnancy in women undergoing assisted reproduction.

We examined the association between progesterone (P4), estradiol (E2), and human chorionic gonadotropin (hCG) levels in early pregnancy and the development of hypertensive diseases of pregnancy among women undergoing assisted reproduction. Retrospective study including patients who underwent frozen embryo transfer (FET), ovarian stimulation (OS), or unassisted conception (UC) and had a live singleton birth. The primary outcome was the development of hypertensive diseases of pregnancy (gestational hypertension, preeclampsia, HELLP, or eclampsia). Secondary outcomes were the development of fetal intrauterine growth restriction (IUGR), gestational diabetes mellitus, birth weight, and pre-term birth. Hormone levels and the development of the outcomes were correlated. A total of 681 patients were included; 189 had FET, 193 had OS, and 299 had UC. Patients undergoing FET or OS were not more likely to develop hypertensive diseases of pregnancy compared with UC patients. While median levels of E2 and P4 were significantly different between P-FET and NC-FET patients (E2: 252 vs 317pg/mL, P4: 64 vs 29ng/mL, respectively; both p < 0.01), rates of hypertensive diseases of pregnancy did not significantly differ between those two groups. In the multivariate analyses, P4, E2, and hCG were not associated with the development of hypertensive diseases of pregnancy, but progesterone levels were significantly higher among those with IUGR. This remained consistent when the analysis was limited to FET patients. P4, E2, and hCG levels did not correlate with the development of hypertensive diseases of pregnancy but elevated progesterone levels did correlate with the development of IUGR.

Preovulatory progesterone levels are the top indicator for ovulation prediction based on machine learning model evaluation: a retrospective study

BackgroundAccurately predicting ovulation timing is critical for women undergoing natural cycle-frozen embryo transfer. However, the precise predicting of the ovulation timing remains challenging due to the lack of consensus among different clinics regarding the definition of this significant event.ObjectiveTo compare the effectiveness of preovulatory serum progesterone levels (P4) versus luteinizing hormone levels (LH) in predicting ovulation time using two machine learning models.Methods771 patients who underwent autologous natural cycle-frozen embryo transfer between January 2015 and February 2022 were recruited. Utilizing variables including follicle diameters, preovulatory serum levels of LH, E2, and P4, two machine learning models were constructed to predict the ovulation time, the importance of the variables in predicting ovulation timing was further ranked.ResultsTwo machine learning models have the capability to accurately predict the timing of ovulation, specifically within 72, 48, or 24 h. The overall accuracy rates of the validation dataset, as determined by the classification trees and random forest models, were found to be 78.83% and 85.28% respectively. Notably, when predicting ovulation within 24 h, the accuracy rate of P4 ≥ 0.65ng/ml exceeded 92%. Furthermore, it was important to consider LH or E2 levels in conjunction with P4 when assessing ovulation timing in cases where P4<0.65ng/ml.ConclusionsPreovulatory serum P4 levels are better predictors of ovulation timing than LH levels and could be used as an alternative in clinical settings, and the model we developed can be used to pinpoint the day of ovulation. Ongoing research and advancements in technology are anticipated to enhance and refine the ovulation method.

The annual ovarian cycle in Northern pit vipers (Bothrops atrox) using serum steroids and ultrasound of the female reproductive tract

Knowledge on hormonal regulation of reproductive cycles in viperid snakes is still incipient, especially when it comes to females and tropical species. There is an urgent need to understand the reproduction of venomous snakes to improve assisted reproduction techniques and optimize the maintenance of these animals in captivity. With this in mind, we monitored Northern pit viper females year-round throughout different seasons via serum levels of progesterone (P4) and estradiol (E2) in conjunction with ultrasound examinations. Ovarian follicles were classified according to their size and stage of vitellogenesis in F-I and F-II (non-vitellogenic phase) or in F-III and F-IV (vitellogenic phase). During autumn and winter, five adult males were rotated among these females for reproductive pairing, which resulted in 17 copulations and 2 pregnancies in the first year and 12 copulations and 5 pregnancies in the second year. Then, we assessed changes in P4 and E2 levels according to seasons, predominant ovarian structures and the presence of embryos or eggs in the oviduct. Our findings showed high levels of E2 when a greater number of vitellogenic follicles were detected, indicating a possible influence of E2 on vitellogenesis and higher levels of P4 whenever eggs and embryos were visualized in the oviduct, implying its role in maintaining pregnancy. Descriptive analysis of the vipers’ ovarian cycles revealed a greater number of vitellogenic follicles during winter, probably as a result of increases in E2; whereas pregnancies occurred predominantly in spring, under the influence of P4. The use of ultrasound images, as a minimally invasive methodology, associated with serum steroid levels has proven to be an efficient approach in the reproductive monitoring of Northern pit vipers in vivo. In addition, these data suggest that female pit vipers under human care display a seasonal reproductive cycle, despite earlier studies involving captive males of the species indicating a lack of seasonality in sperm production and quality.

Integrated Transcriptomic-Metabolomic Analysis Reveals the Effect of Different Light Intensities on Ovarian Development in Chickens.

Light is a key environmental factor regulating reproduction in avians. However, the mechanism of light intensity regulating ovarian development is still unclear. In this study, 5-week-old (5 wk) partridge broiler breeders were randomly divided into a low-light-intensity group (LL group) and a natural-light-intensity group (NL group) (n = 100). In the rearing period (5 wk to 22 wk), the light intensity of the LL group and NL group were 0.41 ± 0.05 lux and 45.39 ± 1.09 lux, and in the laying period (23 wk to 32 wk) they were 23.92 ± 0.06 lux and 66.93 ± 0.76 lux, respectively. Samples were collected on 22 wk and 32 wk. The results showed that the LL group had a later age at first egg and a longer laying period than the NL group. Serum P4 and LH levels in the LL group were higher than in the NL group on 22 wk (p < 0.05). On 32 wk, P4, E2, LH and FSH levels in the LL group were lower than in the NL group (p < 0.05). Ovarian transcriptomics and metabolomics identified 128 differentially expressed genes (DEGs) and 467 differential metabolites (DMs) on 22 wk; 155 DEGs and 531 DMs on 32 wk between two groups. An enrichment analysis of these DEGs and DMs identified key signaling pathways, including steroid hormone biosynthesis, neuroactive ligand-receptor interaction. In these pathways, genes such as CYP21A1, SSTR2, and NPY may regulate the synthesis of metabolites, including tryptamine, triglycerides, and phenylalanine. These genes and metabolites may play a dominant role in the light-intensity regulation of ovarian development and laying performance in broiler breeders.