Serum Progesterone Measurement Research Articles

Maternal Serum Creatine Kinase and Single Serum Progesterone Measurement as Predictors of Ectopic Pregnancy

Objective: To evaluate the role of serum creatine kinase (CK) and progesterone (p) in the prediction of ectopic pregnancy (E.P). Patients and methods: A prospective study was carried out on 60 pregnant women of comparable gestational age and they were divided into 3 groups: Group A: 20 patients with confirmed E.P. Group B: 20 patients with missed or spontaneous miscarriage. Group C: 20 women with normal intrauterine pregnancy (IUP). The following laboratory measurements were performed for the studied groups: 1- Test for serum CK level for all groups. 2- Test for serum progesterone for all groups. 3- Histopathological study to determine the depth of invasion of the trophoblast to the muscular layer of fallopian tube for group A. Results: 1- The serum CK level was significantly higher in group A than in group B and C and was significantly higher in ruptured than in unruptured and in isthmic compared to ampullary EP. 2- The serum progesterone level was significantly lower in group A and B than in group C, but there was no significant difference between group A and B. 3- The serum progesterone level was significantly higher in ruptured than in unruptured EP. 4- There was significant positive correlation between serum CK and progesterone level in group A. Conclusion: 1- An increase in serum CK levels probably antedates tubal rupture and could help in discriminating ruptured from unruptured EP. 2- Serum progesterone discriminates EP from normal IUP but it does not differentiate it from miscarriage. 3- The combination of both serum CK and progesterone in suspected EP has a higher differential accuracy than either analysis alone

Performance characteristics of the light-initiated chemiluminescent assay for quantitative determination of progesterone.

BackgroundTo study the performance of quantitative determination of progesterone by light-initiated chemiluminescent assay (LICA).MethodsClinical samples of serum were used for detection of progesterone by LICA. The precision study was performed according to Clinical and Laboratory Standards Institute (CLSI) EP15-A3, the linear range validation was performed according to CLSI EP06-A, accuracy was evaluated according to CLSI EP9-A3, and the performance of detection capability was confirmed according to CLSI EP17-A2. All data were analyzed using SPSS software. Function regression analysis was performed by OriginPro software.ResultsThe LICA-800 system exhibited low coefficients of variation (CVs) and high reproducibility, and the calculated synthetic CV was 2.16%. The access progesterone assay showed excellent linearity in the assay measuring range (0.37–40 ng/mL) using the polynomial regression method in accordance with CLSI EP06-A. Bias assessment was used to verify accuracy, and the percentage deviation met the quality requirements of the laboratory’s allowable deviation of 10.00%. In terms of the detection capability of LICA, the calculated limit of blank (LoB) was 0.046 ng/mL, limit of detection (LoD) was 0.057 ng/mL, and the limit of quantitation (LoQ) value was 0.161 ng/mL.ConclusionsThe competitive LICA provided a highly sensitive, accurate and precise method for measuring serum progesterone level.

Effect of Ubiquinol supplementation on ovulation induction in Clomiphene Citrate resistance

BackgroundClomiphene Citrate is considered the gold-standard for induction of ovulation and has been used for several years to treat PCOS related infertility. Unfortunately, 15-40% of women with PCOS are resistant to Clomiphene Citrate. The study aimed to evaluate potential benefits of adding the active form of Coenzyme Q10 (Ubiquinol) to Clomiphene Citrate compared with Human Menopausal Gonadotropins (hMG) in Clomiphene Citrate resistant PCOS patients. 148 PCOS Patients with Clomiphene Citrate resistance were randomized into two groups (A and B). In group A, controlled ovarian stimulation was done by Clomiphene Citrate 150 mg daily (from 2nd till 6th day of cycle) together with Ubiquinol starting from 2nd day till day of hCG triggering in a dose of 100 mg orally once daily. In group B, hMG was given from 2nd day of the cycle in a dose ranging from 75 to 225 IU. Serial transvaginal ultrasonography was done starting on cycle day 8 and continued till size of leading follicle reaches 18 mm or more then ovulation triggering was done. Thereafter, patients were advised for a timed intercourse (TI) after 36 hours. A blood sample was withdrawn seven days after hCG triggering, for measurement of serum progesterone. If the Patient presented with a missed period for one week, a serum sample was sent for β-hCG.ResultsThere were no statistically significant differences (P > 0.05) between studied groups regarding; number of cases reaching mature follicular size, number of stimulated cycles, endometrial thickness on the day of hCG triggering, mid-luteal serum progesterone, positive serum pregnancy test and clinical pregnancy rate.ConclusionsAddition of Ubiquinol to Clomiphene Citrate improved ovarian responsiveness in Clomiphene Citrate resistant patients with results comparable to conventional hMG stimulation protocol.

P–615 The effect of late-follicular phase progesterone rise on embryo ploidy, embryo quality and cumulative live birth rates following a freeze-only strategy

Abstract Study question Is late-follicular phase progesterone elevation (PE) associated with a deleterious effect on embryo euploidy, embryo blastulation and cumulative live birth rates (CLBRs)? Summary answer Late-follicular phase PE has no impact on impact on embryo euploidy rate, embryo blastulation rate nor on the CLBR. What is known already The effect of PE in ART outcomes has been extensively studied, yielding so far conflicting results. While some authors claim it is only detrimental to endometrial receptivity, others have suggested that it may also impair oocyte/embryo quality. Moreover, little is known regarding the potential effect PE may have on embryo ploidy and, consequently, CLBR. Study design, size, duration A multicenter retrospective cross-sectional study was performed between August 2017 and December 2019. A total of 1495 ICSI cycles coupled with preimplantation genetic diagnosis for aneuploidies (PGT-A) and deferred frozen embryo transfer (FET) were analyzed. Participants/materials, setting, methods All patients underwent ovarian stimulation with GnRH antagonist protocol and performed a serum progesterone measurement at one of the participating private fertility clinics on the day of trigger. The sample was stratified according to the progesterone levels: normal (≤1.50 ng/ml) and high (>1.50 ng/ml). The primary outcome was the embryo euploidy rate. Secondary outcomes were the number of euploid blastocysts, the blastulation rate and CLBR. Main results and the role of chance Late-follicular phase PE was associated with higher late-follicular estradiol levels (2847.56±1091.10 pg/ml vs. 2240.94± 996.37 pg/ml, p < 0.001) and more oocytes retrieved (17.67±8.86 vs. 12.70±7.00, p < 0.001). The number of euploid embryos was higher in the PE group (2.32±1.74 vs. 1.86±1.42, p < 0.001), whereas the embryo euploidy rate (48.3% [44.9%–51.7%] vs. 49.1% [47.7%–50.6%] and blastulation rate (47.1% [43.7%–50.5%] vs. 51.0% [49.7%–52.4%]) were comparable between the two groups. Likewise, no significant differences were found regarding the live birth rate (LBR) after the first FET (34.1% vs. 31.1%, p = 0.427) nor the CLBRs (38.9% vs. 37.0%, p = 0.637). Mixed-model analysis was performed in order to account for the clustering of cycles in the same patient. Adjusting for patients’ age, PE and BMI, PE failed to demonstrate any effect on the embryo euploidy rate (OR 1.03 [95% CI 0.89–1.20]). Mixed-model analysis for the number of euploid embryos was also performed. After adjusting for PE, age, BMI and ovarian response, PE did not affect the number of euploid embryos (0.02 [95%CI –0.21;0.25]. Multivariate logistic regression adjusted for PE, age, BMI and ovarian response revealed that PE was not associated with the CLBR (adjOR 0.96 [95% CI 0.66–1.38]). Limitations, reasons for caution Limitations of the study include its retrospective nature. Moreover, including only GnRH antagonist protocol and ICSI does not allow the extrapolation of these results to other populations. Wider implications of the findings: Our findings question results from previous studies claiming a detrimental effect of PE on embryo implantation potential. According to our results, PE has no impact on embryo euploidy rate, blastulation rate nor on CLBRs. Trial registration number Not applicable

P–682 Serum progesterone level as prognostic factor in frozen-thawed embryo transfer cycles: effect of selected threshold on gestational results. Systematic review, stratified meta-analysis and meta-regression

Abstract Study question Is there an optimum progesterone threshold level below which gestational results are significantly worse in frozen embryo transfer cycles (FET) with hormone replacement therapy (HRT)? Summary answer Low serum progesterone during luteal phase of HRT-FET cycles impairs substantially its gestational outcomes, regardless of threshold level, origin of oocytes and euploidy of embryos. What is known already HRT for endometrial preparation in FET or oocyte donation cycles is widely used. Oestrogen doses are usually patient-tailored varying upon endometrial thickness, whereas the optimal level of progesterone exposure has not been defined. Various studies have found a negative association between serum progesterone levels measured during luteal phase and FET results in terms of pregnancy and miscarriage rates. Most likely there is an optimal level below which results are worse but a standard threshold level is yet to be established, as in almost every study a different threshold has been found. Study design, size, duration Systematic review and stratified meta-analysis with meta-regression following PRISMA guidelines. An electronic search of MEDLINE, EMBASE, Web of Science, Cochrane Gynaecology and Fertility Specialised Register of Controlled Trials and ClinicalTrials.gov was conducted from inception to January 2021. The aim was to identify prospective or retrospective cohort studies measuring serum progesterone levels around frozen embryo transfer date in HRT cycles. A combination of the following key search terms was used: “progesterone”, “serum”, “frozen embryo”, “transfer”, “frozen-thawed”. Participants/materials, setting, methods Studies analyzing association of luteal serum progesterone with FET-HRT outcomes were included. Risk of bias within studies was assessed using the Newcastle-Ottawa Scale (NOS). Clinical/ongoing pregnancy and miscarriage rates (C/OPR,MR) were considered as primary and secondary outcomes respectively. Odds Ratios with 95% Confidence Interval (OR,95%CI) were calculated applying a random effects model meta-analysis. Heterogeneity was assessed using the I2 statistic. A meta-regression was conducted to examine the association of the effect with the threshold level. Main results and the role of chance The systematic search retrieved 792 studies, 494 after duplicates removal of which 343 were screened and 51 assessed for eligibility. 12 studies, reporting 14 threshold levels, were included in the meta-analysis involving 5009 HRT-FET cycles. Two of them were prospective cohort studies while the rest were retrospective. 10 of them have been published in peer review journals and two were conference abstracts. Quality of studies assessed with NOS varied between 5 and 9. The progesterone threshold ranged from 5.0 to 21.94 ng/ml. Low progesterone levels were associated with less C/OPR (OR: 0.52; 95% CI: 0.40 to 0.66; 11 studies, 5009 cycles). Low progesterone was also associated with high MR (OR: 2.01; 95% CI: 1.57 to 2.58; 9 studies, 2560 pregnancies). These effects showed remarkable consistency in specific sub-analyses considering separately studies with progesterone thresholds up to or above 10 mg/mL, and studies carried out in cycles using oocyte donation, autologous oocytes and embryo aneuploidies screening. Meta-regression did not identify significant association between size effect and progesterone threshold, regarding neither C/OPR (regression coefficient: 0.02; CI 95%: –0.02 to 0.06; p: 0.28) nor MR (regression coefficient: 0.11; CI 95%: –0.13 to 0.36; p: 0.32). Limitations, reasons for caution High degree of clinical and statistical heterogeneity was found due to different routes and doses of progesterone administration, date of progesterone analyses and variety of thresholds as well as high diversity of embryo origin. Despite sensibility analysis by embryo origin any of these sources of heterogeneity can preclude the results. Wider implications of the findings: Despite low progesterone levels are significantly associated to lower gestational results, and a threshold of 10 ng/ml constitutes the median value of our distribution, high quality prospective studies are needed to validate the prognostic value of progesterone levels and to establish an standardised threshold level for clinical application. Trial registration number not required

Gynaecomastia in a male neutered cat with an adrenal tumour and associated hyperprogesteronism, hypercortisolism and hyperaldosteronism.

Case summaryA 7-year-old male neutered domestic longhair cat was presented with chronic progressive gynaecomastia, polydipsia, polyphagia, weight loss and poor fur regrowth. Sexualised behavioural changes were not reported and virilisation was not present on physical examination. Pertinent haematology, biochemistry and urinalysis findings at the time of referral included mild hypokalaemia. Left adrenomegaly and mild prostatomegaly were identified on a CT scan. Evaluation of adrenal hormones with a low-dose dexamethasone suppression test, serum progesterone, testosterone, oestradiol, plasma aldosterone, renin, plasma metanephrine and normetanephrine measurement supported a diagnosis of hyperprogesteronism, hyperaldosteronism and hypercortisolism. Adrenalectomy was performed and histopathology was consistent with an adrenocortical tumour. Clinical signs and hormone elevations resolved postoperatively.Relevance and novel informationTo our knowledge, this is the second report of gynaecomastia secondary to an adrenal tumour in a male neutered cat and the first associated with hyperprogesteronism.

Comparison of a Point-of-Care Analyzer With a Chemiluminescent Immunoassay for Serum Progesterone Measurement in Breeding Management of the Bitch.

Accurate serum progesterone measurements for timing bitches during breeding management is critical for reproductive practice, especially as artificial insemination has become routine to facilitate breeding of animals that are geographically or temporally separated. To measure serum progesterone, chemiluminescent immunoassay (CLIA) has replaced radioimmunoassay as the current standard in the bitch due to its high correlation and increased practicality. In January 2019, a colorimetric point-of-care (POC) immunoassay for quantitative in-clinic canine serum progesterone measurements in <30 min was released. This study provides an independent comparison of the POC (Catalyst One, IDEXX) to the current industry standard, CLIA (Immulite-2000, Siemens). To assess inter-assay imprecision of POC and agreement of the POC and CLIA results, 100 canine serum samples were analyzed on three analyzers (POC-1, POC-2, and CLIA), of which, 74 (POC-1) and 75 (POC-2) results were within POCs' reportable range of 0.2–20 ng/mL and included in the study. To assess intra-assay imprecision, pooled canine serum samples at low (L1), intermediate (L2), and high (L3) progesterone concentrations were analyzed ten times each on POC-1 and CLIA. Relative to CLIA, POC values showed good correlation (POC-1, r2 = 0.9366; POC-2, r2 = 0.9438, P < 0.0001) and significant positive proportional bias at values >2 ng/mL. The POC inter-assay coefficients of variation (CVs) were 13.2% (0.2–2.9 ng/mL, 0.6–9.2 nmol/L, L1), 10.0% (3.0–9.9 ng/mL, 9.5–31.5 nmol/L, L2), 7.1% (10.0–20.0 ng/mL, 31.8–63.6 nmol/L, L3), and 11.2% (all samples). The intra-assay CVs for POC (L1, 15.3%; L2, 7.0%; L3, 4.7%) were higher than those for CLIA (L1, 5.89%; L2, 4.89%; L3, 3.44%). Based on the more rapid increase in serial serum progesterone concentrations in ovulating bitches and the greater imprecision of the POC, the clinical interpretations of serum progesterone measurements as they relate to canine breeding management should be made with caution.

Mifepristone (Korlym ®) Use Interferes With Accurate Serum Measurement of Sex Steroids

Introduction: Mifepristone (MFP) aka RU-486/Korlym is a synthetic steroid analog originally developed in 1980 as an abortifacient that is now used in the management of Cushing’s syndrome (CS). It is both a progesterone (PG) and glucocorticoid (GC) receptor blocker. CS is often associated with reproductive functional anomalies including hypogonadism, menstrual irregularities and infertility. Due to its mode of action, measurement of serum PG and/or cortisol in patients on MFP are inaccurate indices of systemic PG and/or GC deficiency or excess. However, the potential impact of MFP use on other serum steroid assays has not been widely studied. We report the case of a 55 yr old man on treatment with MFP for adrenal CS found to have striking artefactual changes in serum androgen and estrogen levels. Case Summary: A 55 yr old African American man was referred with poorly controlled type 2 diabetes, hypertension, hyperlipidemia, obesity and untreated hypogonadism. He had a 3.2cm left adrenal incidentaloma associated with adrenal CS and he chose medical management rather than surgery. Upon starting MFP at 300mg QD serum total estrogen (by radio-immunoassay; RIA) and estradiol (by chemiluminescence immunoassay; CIA) were markedly elevated while serum total, free, bioavailable testosterone and dihydrotestosterone were all markedly reduced. His HBA1c, weight and energy levels improved on MFP despite these findings. The serum steroid levels normalized to pre-treatment levels after stopping MFP for ~ 4weeks but the changes recurred after restarting therapy. After MFP dose escalation to 300mg BID the serum steroid levels normalized after stopping MFP for ~ 6 weeks. The artefactual low testosterone levels also occured with measurement by equilibrium dialysis but “normal accurate” results were obtained when measured by liquid chromatography-Tandem mass spectrometry (LC-TMS). He remains on MFP 300mg BID without need for androgen repletion. Discussion: With increased use of MFP for CS, indices for tracking its clinical and biochemical effects assume great importance. There are few reports of the possible effects of MFP on estrogen and testosterone serum assays despite its touted low cross reactivity with sex steroids. Our case suggests that the significance, extent and prevalence of artefactual changes on serum sex steroid assays may be underestimated and under- appreciated. Conclusions: Our case of wide disparities in serum estrogen and androgen measures in a patient on MFP indicates that caution needs to be exercised in the interpretation of such results in patients on current MFP therapy. Our clinical observations suggest depending on the dose that a wash out period of 4–6 weeks is required to ensure accurate measures. Studies to ascertain the prevalence of this artefactual effect are needed and it appears testosterone measurement by LC-TMS obviates the testosterone assay artefact.

Endometrial compaction and serum progesterone measurements at the day of embryo transfer cannot predict pregnancy outcomes in frozen-thaw embryo transfer cycles

Endometrial receptivity plays a basic role in successful embryo implantation and pregnancy outcomes and can be assessed by many of non-invasive markers. Our study evaluated the impact of two of these markers specifically serum progesterone and endometrial thickness at embryo transfer day in prediction pregnancy outcomes on (60) patients attempting medicated frozen embryo transfer (FET) cycles. All patients were received sequential estrogen &amp; progesterone medications for endometrial preparation then submitted to measurements of endometrial thickness (EMT) by transvaginal-ultrasound (TV-US) &amp; serums progesterone (P) analysis at the embryo transfer day, thereafter day 3 verified-thawed embryos grades (A±B) were transferred. Compacted (decreased) EMT was seen in 48.3% of patients with higher pregnancy rate (PR) of 58.6%t than non-compacted EMT (no change or increased) which was seen in 51.7% of patients with (PR) of 29.0%, (P value=0.021). However ongoing pregnancy rate (Ong PR) not differed significantly between both groups (44.8% in compacted vs 25.8% in non-compacted, P value=0.053), also the means of serum P not differed between pregnant and non-pregnant patients (P value=0.374). ROC curves for Ong PR prediction in relations to endometrial compaction &amp; serum progesterone at embryo transfer day were poor (AUC= 0.630, &amp; AUC=0.576, respectively). This study suggested that endometrial compaction or serum P levels measurements at embryo transfer day were poor predictors for ongoing pregnancy where any kind of EMT changes (decreased or not) seen after P administration not significantly affect pregnancy outcomes in frozen-thaw cycles of cleavage stage embryos transfer.

The effect of various administration routs of D-cloprostenol at the time of artificial insemination on conception rate in dairy cattle

The aim of the present study was to evaluate the effect of various administration routes of PGF2α analogue, D-cloprostenol, at the time of artificial insemination (AI) on conception rate in a large dairy herd. A total number of 659 lactating dairy cows (101 heifers and 558 cows) were enrolled in the study. Following heat detection, the cows were artificially inseminated according to the AM-PM rule. The animals were divided into a control (29 heifers and178 cows) and three experimental groups; IM (53 heifers and 110 cows), IV (155 cows) and IU (19 heifers and115 cows) groups that received intramuscular 150 µg, intravenous 150 µg and intrauterine 5 µg of D-cloprostenol, respectively, at the time of AI. The control group did not receive any treatment at the time of AI. To measure serum progesterone concentration, blood samples were obtained from 15 cows in each group at the time of insemination and on days 4 and 15 post-insemination. Pregnancy was diagnosed by rectal ultrasound examination between days 35-40 post-insemination. The effects of treatments, parity, milk production, and days in milk (DIM) on conception rate of adult cows were evaluated using the multivariate logistic regression model. Intramuscular administration of D-cloprostenol at the time of AI significantly increased conception rate in dairy cows compared to the control group (60.1% vs. 40.6%; p = .005). However, intravenous and intrauterine administration of D-cloprostenol did not alter the conception rate (32.3%; p = .29 and 50%; p = .11, respectively). D-cloprostenol treatment at AI could not significantly increase the conception rate of heifers followingboth (IM & IU) administration routes. In conclusion, this study showed that intramuscular administration of D-cloprostenol at the time of insemination might improve the conception rate in adult cows, while its intravenous and intrauterine administration did not induce any positive effects on reproductive efficiency of dairy cows. HIGHLIGHTS Intramuscular administration of D-cloprostenol at the time of insemination might improve the conception rate in adult cows. Intravenous administration of D-cloprostenol did not induce any positive effects on reproductive efficiency of dairy cows. Intrauterine administration of D-cloprostenol did not induce any positive effects on conception rate of dairy cows.

Can the pregnancy outcome be improved by incrementing progesterone supplementation on the day of embryo transfer in women undergoing fresh and frozen embryo transfer cycles?

Context: Luteal phase defect has been identified in all assisted reproductive technology cycles, necessitating optimal progesterone levels during the window of implantation (WOI) to improve reproductive outcomes. Aims: The aim of this study was to determine if incrementing progesterone supplementation based on the levels of serum progesterone(P) measured on the day of embryo transfer (ET) has an impact on the pregnancy outcome in fresh and frozen ET cycles. Settings and Design: A total of 220 women undergoing fresh and frozen ET were prospectively enrolled and received standard luteal phase support, an increment in P supplementation made if serum P levels were less than 15 ng/mL. Materials and Methods: Serum progesterone levels were assessed for all the patients on the day of ET. Patients undergoing fresh and frozen ETs were divided into two groups based on serum progesterone levels on the day of ET. If serum progesterone levels were found to be less than 15 ng/mL on the day of ET, the dose of progesterone supplementation was incremented. A correlation between the groups with serum progesterone less than and more than 15 ng/mL on the day of ET with the pregnancy outcome was then made. Results: No statistically significant difference was observed in the clinical pregnancy rate and biochemical pregnancies in the groups with serum progesterone levels less than or more than 15 ng/mL on the day of ET when an increment in the progesterone supplementation was performed (both fresh, P = 0.35 and frozen cycles, P = 0.056). Statistical Analysis Used: Chi-square test was used for qualitative analysis, and the Student’s t test was used for comparison of means. Conclusions: Measuring serum progesterone levels on the day of ET and incrementing P supplementation if serum levels of progesterone are less than 15 ng/mL to obtain optimal progesterone levels during the WOI in both fresh and frozen ET cycles might help in improving our pregnancy outcomes.

Neuro-pharmacological reinstatement of ovulation and associated neurobiology in a macaque model of functional hypothalamic amenorrhoea.

What is the underlying neuropathology in a cynomolgus macaque model of functional hypothalamic amenorrhoea (FHA) and can it be normalized to restore ovulation? Anovulatory monkeys exhibited increased hypothalamic norepinephrine (NE), kisspeptin and gonadotropin-releasing hormone (GnRH) in the early follicular phase, but administration of the NE reuptake inhibitor (NRI), reboxetine (REB), restored ovulation during stress and normalized NE, kisspeptin and GnRH. Female cynomolgus macaques, like women, show individual reproductive sensitivity to modest psychosocial and metabolic stress. During stress, resilient females ovulate through two menstrual cycles whereas stress-sensitive (SS) macaques immediately cease ovulation. On Day 5 of a non-stressed menstrual cycle, resilient macaques have less NE synthesizing enzyme [dopamine β-hydroxylase (DBH)], kisspeptin and GnRH innervation of the medial basal hypothalamus but more endogenous serotonin than SS macaques. Stress increased DBH/NE, kisspeptin and GnRH but did not alter serotonin. In a longitudinal design, 27 adult (7-13 years) female cynomolgus macaques (Macaca fascicularis) with three different levels of sensitivity to stress were monitored with daily vaginal swabs and frequent serum progesterone (P) measurements. Three 90-day experimental periods called 'Cycle Sets' were monitored. A Cycle Set consisted of one ovulatory menstrual cycle without stress, and two cycles, or 60 days, with modest stress. Each Cycle Set was followed by a rest period. During a Cycle Set, individuals were either untreated (placebo) or administered escitalopram (CIT) or REB. Ultimately, half of each sensitivity group was euthanized during stress with CIT or REB treatment and the hypothalamus was obtained. Neurobiological endpoints were compared between CIT and REB treatment groups in stress resilient and SS monkeys. The monkeys were housed at the University of Pittsburgh primate facility for the duration of the experiments. Upon euthanasia, their brains and serum samples were shipped to the Oregon National Primate Research Center. The hypothalamus was examined with immunohistochemistry for the expression of DBH (a marker for NE axons), kisspeptin and GnRH. P was measured in the serum samples by radioimmunoassay. Daily administration of REB restored ovulation in 9 of 10 SS animals during stress. Of note, REB significantly increased P secretion during stress in the most sensitive group (P = 0.032), which indicates ovulation. CIT lacked efficacy. REB significantly reduced DBH/NE, kisspeptin and GnRH axon density in the hypothalamus relative to CIT treatment (P = 0.003. 0.018 and 0.0001, respectively) on Day 5 of the menstrual cycle in resilient and sensitive groups. N/A. The US FDA has not approved REB for human use, although it is used in Europe for the treatment of depression/anxiety as EdronaxTR. Whether REB could be useful for the treatment of FHA in women has not been determined. The use of an NRI to treat FHA is a novel approach and the potential reinstatement of ovulation could be straightforward compared to current treatment protocols. The underlying neurobiology provides a compelling case for treating the origin of the pathology, i.e. elevated NE, rather than circumventing the hypothalamus altogether with gonadotropins, which have associated risks such as hyperstimulation syndrome or multiple births. Portions of this study were supported by NIH grant HD062864 to C.L.B., NIH grant HD62618 to J.L.C. and C.L.B. and 1P51 OD011092 for the operation of the Oregon National Primate Research Center. There were no competing interests.

Revisiting the role of serum progesterone as a test of ovulation in eumenorrheic subfertile women: a prospective diagnostic accuracy study

To estimate the prevalence of ovulatory cycles in eumenorrheic subfertile women and compare the diagnostic accuracy of a single ultrasound with serum midluteal progesterone measurement in detecting ovulatory cycles. Prospective diagnostic accuracy study. University-level hospital. A total of 208 subfertile eumenorrheic women. None. To estimate the prevalence of ovulatory cycles in eumenorrheic women and compare the diagnostic accuracy of a single, well-timed ultrasound scan (index test) with serum progesterone measurement (reference test) by calculating the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and positive likelihood ratio (LR+) and negative likelihood ratio (LR-). The prevalence of ovulatory cycles among subfertile eumenorrheic women was 92.9% and 99.5% when midluteal serum progesterone level and ultrasound scanning were used as the reference test, respectively. The sensitivity, specificity, PPV, and NPV of ultrasound in identifying ovulatory cycles were 100%, 7.1%, 93.4%, and 100%, respectively. The LR+ and LR- were 1.1 and 0, respectively. The agreement between the ultrasound and serum progesterone was almost perfect (prevalence and bias-adjusted kappa = 0.81. The prevalence of ovulatory cycles in eumenorrheic subfertile women appears to be high. A single well-timed ultrasound can be performed to identify ovulatory cycles during the infertility workup in eumenorrheic women.

Use of oxytocin to attain cervical dilation for transcervical embryo transfer in sheep.

The aim of this work was to determine whether a cervical dilation protocol (CDP) composed of only oxytocin can be used to perform transcervical (non-surgical) embryo transfer in sheep (NSET) without affecting the viability of the corpus luteum (CL). Likewise, we evaluated whether a cervical transposing test with a Hegar dilator (CT Hegar test), performed at oestrous time, could be used to screen ewes for NSET (greater or lower chances to transpose the cervix). For that, oestrous and ovulation synchronization was performed in 25 Santa Inês ewes to induce the dioestrous condition. Animals went through the following CDP in a crossover design: E+OX, oestradiol benzoate (100µg intravenously [IV]) and oxytocin (100IU IV); OX, oxytocin (100IU IV); and SAL, saline solution (IV). Using a Hegar dilator, cervical transposing attempts were performed at oestrous (D0) and dioestrous time (D8). The viability of the CL (morphology, luteal blood flow and progesterone values) was evaluated by ultrasonography (colour Doppler and B-mode) and by serum progesterone measurement from D7 to D13. The cervical transposing rate was lower for the SAL group (64%; 16/25; p<.05) and did not differ between the E+OX (88%; 22/25, p>.05) and OX (84%; 21/25, p>.05) groups. No treatment affected the CL viability. The CT Hegar test showed a high sensitivity (85.7%-93.3%), satisfactory accuracy (72%-84%), low false-negative rate (6.7%-14.6%), but high false-positive rate (46%-66.7%). In conclusion, a CDP protocol composed exclusively of oxytocin can lead to good cervical transposing rates and does not affect the viability of the CL. In addition, a screening test (CT Hegar) performed at oestrus can identify ewes for which cervical transposing will likely not occur at NSET.

The efficacy of vitamin D combined with clomiphene citrate in ovulation induction in overweight women with polycystic ovary syndrome: a double blind, randomized clinical trial.

The aim of this study was to assess the effect of vitamin D supplementation on ovulation rate in overweight subfertile women with PCOS undergoing ovulation induction. This was a single center, parallel-groups, double-blind, and placebo-controlled randomized trial involving 186 eligible women undergoing induction of ovulation with clomiphene citrate (Clomid®, Aventis) 50 mg tablet twice daily starting from the third day of menstrual cycle and for 5 days combined with either oral Vitamin D (ossofortin®, EVA PHARMA) 10,000 IU twice weekly and calcium (calciprex®, Marcyrl Pharmaceutical Industries) 1250 mg twice daily or to receive a placebo with calcium for three successive induction cycles. The vitamin D or placebo supplementation started 1 month before induction cycles (total four cycles). Cycles were monitored with ultrasound follicle tracking and mid-luteal serum progesterone measurement. The primary outcome was the ovulation rate after three induction cycles. The study was performed during the period between January 2018 and September 2018, Eighty six (92.5%) women in the treatment group and 73 (78.5%) in the control group had successful ovulation (p = 0.007). The absolute and relative risk reduction was 14% and 65% respectively. Biochemical and clinical pregnancy occurred in 61.3 and 50.5% in the treatment group, and in 49.5 and 39.8% in the control group (p = 0.105 and 0.141 respectively). In subfertile women with PCOS undergoing induction of ovulation, vitamin D supplementation significantly improved the ovulation rate; however, there was no effect on clinical or biochemical pregnancy.

Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients.

ObjectiveThe aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows.MethodsTwelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination.ResultsThe findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12).ConclusionIt may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

The Ability of a Single Serum Progesterone Measurement to Predict the Prognosis of First Trimester Pregnancy

OBJECTIVE: There are cases where ultrasonographic evaluation is inadequate in determining first-trimester healthy pregnancy. In these cases, both the patient and the clinician are involved in a worrying expectation. The single serum progesterone measurement was tried to predict pregnancy prognosis in short time period. This study evaluated the predictive value of a single serum progesterone measurement in first-trimester pregnancy loss.STUDY DESIGN: A prospective case-control study was conducted between July 2013 and July 2014 in Tertiary Hospital. The study compared 106 patients with threatened abortion (vaginal bleeding or spotting) before 13 weeks of gestation with 110 healthy control pregnancies. Samples of serum progesterone were obtained on the day each patient was admitted to the hospital. A Beckman-Coulter macro-Enzyme-linked immunoassay (ELISA) system was used to measure serum levels of progesterone (Beckman-Coulter, Ireland).RESULTS: Progesterone levels were measured when the patients first visited the hospital and they were followed until the end of the first trimester. Progesterone levels differed significantly (p=0.013) between the two groups. The spontaneous abortion rate was significantly (p=0.044) higher in the case group. Progesterone levels differed significantly (p=0.001) between ongoing pregnancy and spontaneous abortion groups. Using a cut-off level of 10.7 ng/mL, the sensitivity, specificity, and positive and negative predictive values were 92.1%, 44%, 92.6%, and 42.3%, respectively, for viable pregnancies.CONCLUSION: A single measurement of progesterone levels in serum can predict the prognosis of a viable first-trimester pregnancy, while it is less effective for identifying non-viable pregnancies.

Serum progesterone levels could predict diagnosis, completion and complications of miscarriage

BackgroundLow serum progesterone levels were strongly correlated with miscarriages in several publications and with completion of miscarriage in one paper. This study evaluated several parameters, predominantly serum progesterone, as predictors for miscarriages, their swift non-surgical completion and their complications. Basic proceduresSuspected or confirmed non-viable pregnancies with available concomitant serum progesterone measurements were retrospectively reviewed. The performance of serum progesterone, either alone or combined with other parameters, to predict viability, surgical removal and delay of non-surgical evacuation of non-viable pregnancy and complications, was analysed by logistic regression combined with Akaike and Bayesian information criteria, likelihood, receiver operated characteristic (ROC) curves, Mann-Whitney test and Fisher’s exact test. Main findingsFrom 151 included pregnancies, 104 (68.9 %) were non-viable with 91 completions of miscarriage without surgery. The probability of viability was correlated linearly and curvilinearly with serum progesterone (p < 0.001). The probability of surgical removal, and the delay before non-surgical evacuation, showed a linear relationship with progesterone. No complication occurred when progesterone levels remained below 10 μg/L, while its rates were 9.5 % of non-viable pregnancies with progesterone levels between 10 and 20 μg/L and 26.7 % of cases with progesterone levels above 20 μg/L. Combined with progesterone, either “parity” or “history of miscarriage” improved the prediction of viability, “history of supra-isthmic uterine surgery” improved the prediction of surgery and “history of miscarriage” improved the prediction of delayed non-surgical evacuations. ConclusionSerum progesterone can probably predict the odds of miscarriages, surgical removal, delayed non-surgical evacuation and complications, with potential improvements when different predictors are combined.

Can combining vaginal and rectal progesterone achieve the optimum progesterone range required for implantation in the HRT-FET model?

What is the ongoing pregnancy rate (OPR) in frozen embryo transfer (FET) cycles, using combined rectal and vaginal progesterone in hormonal replacement therapy (HRT)? A prospective cohort study (n = 277) including 239 HRT-FET cycles with serum progesterone measurements studying combined vaginal (90 mg/12 h) and rectal (90 mg/12 h) progesterone administration and single blastocyst transfer on the sixth day of progesterone administration. A total of 134 responses to questionnaires covering convenience and side-effects were collected. The median serum progesterone level was 45 nmol/l (range 2-150 nmol/l). Overall positive HCG rate, OPR at week 12 and pregnancy loss rates were 62%, 44% and 29%, respectively. A non-linear relationship between serum progesterone levels and OPR was found. Crude odds ratio for OPR in the high progesterone group (>45 nmol/l) was 0.56 (95% CI 0.32 to 0.98; P = 0.04) compared with the intermediate progesterone group (28-45 nmol/l). Discomfort after rectal progesterone administration was reported on the embryo transfer day and on the day of pregnancy scan 5 weeks later by a total of 18% (16/87) and 17% (8/47) of patients, respectively. Discomfort related to vaginal administration increased significantly over time and was reported by 18% (16/87) on the day of embryo transfer compared with 45% (21/47) on the day of pregnancy scan (P < 0.002). Combined rectal and vaginal progesterone in HRT-FET cycles resulted in higher median progesterone levels compared with vaginal administration alone. This study suggests that an upper threshold for serum progesterone exists and that above this concentration serum progesterone levels decrease the OPR. Rectally administered progesterone was well tolerated by patients.

Quantitative analysis of progesterone using isotope dilution-matrix-assisted laser desorption ionization-time of flight mass spectrometry as a reference procedure for radioimmunoassay

BackgroundProgesterone is one of the female steroid hormones and plays an important role in the menstrual cycle and during pregnancy. It is especially important in preparing the uterus for the implantation of the blastocyst and maintaining pregnancy. The concentration in human serum is measured to determine the ovarian function retroactively and the cause of abortion in early pregnancy. MethodsA quantification assay based on isotope dilution mass spectrometry to determine the concentration of progesterone in human serum is reported. Incorporated with 13C3-progesterone, serum samples were subjected to progesterone extraction and clean-up by C4 solid-phase-extraction columns and hexane-based liquid/liquid extraction, respectively. The cleaned-up serum samples were then subjected to MALDI-TOF mass spectrometry for the quantification of progesterone. ResultsProgesterone and the internal standard, 13C3-progesterone, were measured in the selected reaction monitoring mode for the transitions m/z 315.4 to 108.9 and m/z 318.4 to 111.9, respectively. We calculated the peak area ratio of progesterone to 13C3-progesterone. The progesterone concentration in human serum was calculated by substituting the peak area ratio into an isotope dilution calibration curve, and then compared with the radioimmunoassay. ConclusionsIn the study, the concentrations of serum progesterone were measured, and the recovered progesterone concentration determined by the assay showed good robustness and consistency in comparison to the conventional radioimmunologic assay. We concluded that the 13C3-progesterone-based quantification assay is a robust method for the measurement of serum progesterone.