Early Luteal Phase Research Articles

P-305 Uterine contractile function across the menstrual cycle in healthy women: an exploration of objective reference values of sub-endometrial motion using speckle tracking

Abstract Study question To explore the characteristics of normal uterine contractile function across the menstrual cycle in healthy women using a novel quantitative ultrasound method. Summary answer We confirm reproducible differences in uterine contractility across the menstrual cycle; highest activity seen in the periovulatory phase, and lowest in the late luteal phase. What is known already Uterine peristalsis is the rhythmic, wave-like motion of the subendometrial layer of the uterus. Various subjective methods using visual interpretation suggest that uterine peristalsis features are different in the various stages of the menstrual cycle, and they are thought to be important for fertility. However, no method has been developed which can assess all uterine contraction features simultaneously. Recently, a new automated quantitative method to measure uterine contractility was validated in IVF patients to analyse uterine peristalsis on transvaginal ultrasound recordings with speckle-tracking. With this method a new contraction feature – coordination – can be assessed alongside frequency, direction, amplitude. Study design, size, duration Multi-centre observational prospective cohort study carried out in the outpatient gynaecology department of the Catharina Hospital in Eindhoven, the Netherlands, the University of Naples, Federico II, Naples, Italy and Embryolab Fertility centre in Thessaloniki, Greece. Patients were included from September 2020 up to January 2022. Primary outcomes were the contraction frequency (contractions/minute), amplitude, direction (Cervix-to-fundus, Fundus-to-cervix), and coordination. Participants/materials, setting, methods 63 women were included from the gynaecological department of participating centres. Women were included if they were ≥18 years of age, premenopausal and had a normal, natural menstrual cycle. A normal cycle was defined as: regular (duration ± 28 days), no dysmenorrhea, no menometrorrhagia. Transvaginal ultrasounds were performed during the menstrual phase (M), late follicular (periovulatory) phase (LF), early luteal phase (EL, ovulation + 3 days) and/or late luteal phase (LL, ovulation + 7 days). Main results and the role of chance Uterine contractility features were extracted from the gathered ultrasound recordings using a quantitative dedicated speckle tracking algorithm previously developed by our group. Each patient underwent a four-minute ultrasound of the uterus in mid-sagittal section. Patients had a mean age of 32 years, cycle duration of 27.8, and BMI of 23.0. The majority of women was nulliparous. The majority of uterine contraction features differed significantly between menstrual cycle phases. Contraction frequency was highest in the LF phase and lowest in the M and LL phases (1.55 vs. 1.28/min, p = 0.001). A trend was found for contractions during the periovulatory and early luteal phases contractions to travel mainly from cervix-to-fundus direction, whilst the menstrual phase showed contractions mainly from the fundus to cervix (p > 0.05). No significant difference between phases was found for contraction amplitude. Contraction coordination (simultaneous contraction of the anterior and posterior walls in the same direction) was highest in the periovulatory phase (p = 0.002). Our results are in line with with previous published studies using subjective visual inspection. Limitations, reasons for caution This is the first study investigating contraction coordination as a specific feature of uterine peristalsis. Not all patients were available for multiple ultrasounds during the same cycle, thereby a within-subjects comparison of the uterine contractions was not possible. Wider implications of the findings We explored further implementation of a quantitative method of uterine peristalsis measurement. Our investigation of a novel contraction characteristic - coordination - opens up possibilities for research and a potential etiological/treatment target. Further research using this method and suggested reference values will provide insight into contraction characteristics in abnormal uteri. Trial registration number NCT02310802

P-366 Tumor necrosis factor-alpha and milk fat globule-epidermal growth factor 8: novel biomarkers to predict implantation failure and pregnancy loss

Abstract Study question Can implantation failure (IF) and pregnancy loss be predicted in serum prior to in vitro fertilization (IVF)? Summary answer Tumor necrosis factor (TNFα) and milk fat globule-epidermal growth factor 8 (MFG-E8) levels may serve as serum markers to predict IF and pregnancy loss. What is known already In a normal pregnancy, mediators such as TNFα are released creating a physiological inflammatory response. However, an exaggerated release of TNFα has been associated with IF and recurrent pregnancy loss (RPL). Recent studies demonstrated that TNFα up-regulates the expression of inflammatory factors such as MFG-E8. MFG-E8 is known to modulate implantation by acting at various levels of the trophoblast and endometrial compartments. Hence an overexpression of this protein may result in apoptosis, endometrial damage, and impaired implantation. Study design, size, duration This multicentric prospective controlled pilot clinical study was conducted from January 2016 to January 2020 and included 30 women in their natural cycle in which serum MFG-E8, TNFα, estradiol (E2), and progesterone (P4) levels were quantified in the early proliferative (cycle day 2) and secretory phases (urinary LH + 7 days). Additionally, an endometrial biopsy was obtained on urinary LH + 7 days for MFG-E8 and TNF α protein and gene expression analysis. Participants/materials, setting, methods Women ages 21-35y were recruited from 3 groups: fertile controls (C), unexplained IF (following 3 failed good quality embryo transfers), and RPL (at least 2 unexplained first trimester miscarriages). Patients with history of uterine surgery, abnormal uterine cavity (fibroids, endometrial polyps, adhesions, adenomyosis, and congenital uterine abnormalities), hydrosalpinx, diminished ovarian reserve, harboring chromosomal rearrangements, thrombophilia, or autoimmune diseases were excluded. Main results and the role of chance Ten women were included in each group. No statistical differences were found in age, BMI, AMH, baseline FSH, and baseline antral follicle count among cohorts. Mean serum E2 and P4 levels were similar among all groups in both the proliferative and secretory phases: E2 proliferative (C 69.19±26.64 pg/ml, IF 64.19±32.56 pg/ml, RPL proliferative 57.44±38.51; p = 0.55), E2 secretory (C 164.10±52.57 pg/ml, IF 172.57±121, RPL 173.81±.97.35; p = 0.25), P4 proliferative (C 0.45±0.15 ng/ml, IF 0.45±0.19 ng/ml, RPL 0.53±0.18 ng/ml; p = 0.85), P4 secretory (C 7.42±4.06 ng/ml, IF 7.8±4.56 ng/ml, RPL 8.05±4.38 ng/ml; p = 0.74). Mean serum TNFα levels were significantly higher in both, the proliferative and secretory phases for the RPL group (proliferative RPL 9.98±4.47 pg/ml, IF 4.73±2.56 pg/ml, C 3.42±1.01 pg/ml; p = 0.001 vs secretory RPL 8.67±4.45 pg/ml, C 3.35±0.94 pg/ml, IF 3.85±1.01 pg/ml; p = 0.03). Mean serum MFG-E8 levels were significantly higher in the IF group during the proliferative phase (IF 373±201 pg/ml, RPL 201±115 pg/ml, C 225.58±109.73pg/ml; p = 0.03), but not in the secretory phase (IF 237±101 pg/ml, RPL 189±116 pg/ml, C 199.41±112.43 pg/ml; p = 0.15). Endometrial MFG-E8 mRNA levels were significantly lower in the IF and RPL group compared to C (p = 0.03). TNFα mRNA levels were not statistically significant among groups (p = 0.12). Limitations, reasons for caution This is a pilot study to assess feasibility. Due to the small sample size, the effects of more subtle covariates would not have been detected. Future larger studies are warranted. Wider implications of the findings These novels differentially expressed serum and endometrial markers may provide information on the physiology of implantation and could generate the basis for non-invasive diagnostic tools and therapeutic use of MFG-E8/TNFα antagonists in women with IF and RPL. Trial registration number IIT-2014-100366

Effects of the menstrual cycle on cytological false-negatives in women with persistent cervical intraepithelial neoplasia.

False-negatives on cytology may be observed during follow-ups for patients with persistent cervical intraepithelial neoplasia (CIN); however, the underlying reasons are unknown, and the relationship between the intra-individual variability of false-negatives and the menstrual cycle phase has not been elucidated. Therefore, this study aimed to determine whether the menstrual cycle influences cytological results to maximise the accuracy of such tests. A total of 154 liquid-based cytological (LBC) samples were obtained during follow-ups for 26 patients with CIN, and the relationship between cytological results and the menstrual cycle, which was classified into six phases, was analysed. All LBC smears were satisfactory, and 20 of 154 (13.0%) specimens were negative for intraepithelial lesions or malignancy (NILM). A statistically significant association was observed between the cytological results and the phase of the menstrual cycle, χ2 (2)=19.322, P < 0.01. The association was moderately strong (Cramer's V=0.354). The value of adjusted residuals showed a statistically significant increase in the NILM percentage as a cytological result during the early secretory phase (P < 0.01) and a statistically significant decrease in NILM during the menstrual and proliferative phases (P < 0.01). The present study revealed that false-negative cytological results were found to more likely to occur during the early secretory phase. More careful and precise microscopic observation of Pap smears collected at the early secretory phase may contribute to a reduction in the occurrence of false-negatives and improve cytological sensitivity.

Early and Late Luteal Executive Function, Cognitive and Somatic Symptoms, and Emotional Regulation of Women with Premenstrual Dysphoric Disorder.

Objective: Cognitive and somatic symptoms were vital factors in developing personalized treatment of depressive disorder. The study aimed to evaluate the following: (1) the cognitive and somatic symptoms of premenstrual dysphoric disorder (PMDD) in the early luteal (EL) and later luteal (LL) phase; and (2) their association with depression and functional impairment of PMDD. Methods: We prospectively evaluated executive function, emotion regulation, cognitive and somatic symptoms, and depression in the EL and LL phases in women with PMDD. Sixty-three women with PMDD and 53 healthy controls completed Simon’s task and questionnaire to assess emotion regulation, inattention, fatigue, insomnia, and depression. Results: Women with PMDD had a poor performance in Simon’s task during the LL phase. They were less likely to exercise cognitive reappraisal during EL and LL phases. Their cognitive reappraisal positively correlated with executive function and negatively associated with depression. In the LL phase, they also experience higher inattention, insomnia, and fatigue, which correlate with the depression and functional impairment of PMDD. Inattention is the most associated factor of PMDD and functional impairment in controlling depression. Conclusion: Executive function was impaired in women with PMDD during the LL phase. Its performance correlated positively with emotion regulation and negatively with depression. The association between inattention and PMDD functional impairment indicates that evaluation and intervention for cognitive impairment were essential in treating women with PMDD. Further studies were required to elucidate the possible etiology underlying these associations.

Evolution of serum progesterone levels in the very early luteal phase of stimulated IVF/ICSI cycles post hCG trigger: a proof of concept study.

Studies have suggested that controlled ovarian hyperstimulation adversely affects endometrial receptivity due to advanced endometrial maturation. This adverse effect is mainly attributed to supraphysiological levels of both estrogen and progesterone identified in stimulated cycles. There is a paucity of published data investigating the very early luteal steroid profile following hCG trigger. This prospective, observational study was undertaken to determine the increase in serum progesterone levels after human chorionic gonadotrophin (hCG) trigger in stimulated IVF/ICSI cycles. This proof-of-concept study included 11 patients requiring ovarian stimulation for IVF/ICSI and who planned to avail of pre-implantation genetic screening with embryo vitrification of their biopsied embryos at blastocyst stage. For each study participant, five additional blood samples were drawn at the following specific times in the stimulation cycle, on the morning (10.00-12.00) of the assigned day to induce final oocyte maturation with hCG trigger, immediately prior to administration of hCG for final oocyte maturation, 1h, 2h, and 36h post hCG trigger. A prediction model, the Gompertz curve, was used to determine serum progesterone levels at intervals between the 2h post hCG trigger sample and the day of oocyte retrieval. Statistically significant increases in serum progesterone levels were identified following hCG administration as early as 1h following trigger (P4 0.57ng/ml, p < 0.05), 2h following trigger (P4 0.88ng/ml, p < 0.001) and on the day of oocyte retrieval (P4 9.68ng/ml, p < 0.001). According to our prediction model, the Gompertz curve, the projected serum progesterone level at 4h post trigger would have achieved a level of 1.45ng/ml, 8h post trigger of 3.04ng/ml, and 12h post trigger of 4.8ng/ml. The very early and significant increases in serum progesterone following hCG trigger are clearly demonstrated in this study. The endometrium is undoubtedly exposed to rapidly increasing serum progesterone levels post hCG trigger that would not be identified until much later in natural menstrual cycles. This study is registered with clinicaltrials.gov under the identifier NCT04417569.

Luteal haemodynamics during spontaneous and induced luteolysis in crossbred cattle

The haemodynamics of CL during natural and induced luteolysis was studied in crossbred cattle using colour Doppler ultrasonography. In natural cycle, penetration of Doppler signals deep into the CL was observed during Day 15 – 17 coinciding with the initiation of luteolysis. When PG was administered in the early luteal phase (Day 4), there was an increase in blood flow intensity in the periphery of CL, but when PG was administered in the mid luteal phase (Day 10), the intensified Doppler signals penetrated deep into the luteal parenchyma within two hours. There were no significant differences in velocity parameters and RI values between the treatment groups. Thus it was concluded that the characteristic difference in the intensity and nature of vascular perfusion was the determining factor for the initiation of luteolytic process in cattle.

Is I-shaped uterus more common in patients with hyperandrogenism?

ObjectivesSubtypes of T-shaped uterus are rare uterine cavity anomalies and there are no morphometric criteria for the diagnosis. Earlier we established a high frequency of I-shaped uterus in patients with adrenalhyperandrogenism, which is more common in Armenian populations. The aim of the study was to determine the frequency of I-shaped uterus as a subtype of T-shaped uterus in patients with ovarian and adrenal hyperandrogenism, accompanied by infertility and miscarriage, as well as the development of it's ultrasonic morphometric criteria. Study designWe conducted an ultrasound of 486 patients aged 19–40 years (mean 30.1 ± 5.5) who applied for infertility or habitual pregnancy loss.74 of them were diagnosed with the PCOS (Polycystic ovary syndrome) and 43-CAH (congenital adrenal hyperplasia). Ultrasound was performed in early luteal phase. The classification of uterine cavities was carried out according ESHRE/ESGE. Results299 had normal ultrasound morphology of the uterine cavity, 20.7% various uterine cavity abnormalities. T-shaped uterus was observed in 3.7%, I-shaped uterus exclusively in patients with hyperandrogenism, 24.3% with PCOS and 39.5% with CAH. To determine the relevant morphometric features as diagnostic criteria for the I-shaped uterus some measurements were performed. The values of dist1-dist2 and dist1-dist3 in the normal cavity had a significant difference (P1-2 0.3), also the cavity width in the middle third and the isthmic section did not have a significant difference (P > 0.05), while in the normal cavity shape these values were significantly different (P < 0.05). ConclusionFrequency of occurrence of the T-shaped uterus did not exceed that in comparison with a group of women with other causes of infertility, while I-shaped congenital anomaly of the uterine cavity was found in 24.3–39.5% patients with hyperandrogenism. The difference between the interostial and corporal distances and the interostial and isthmic distances was the most relevant morphometric attribute of I-shaped uterus.

Good things come to those who mate: analysis of the mating behaviour in the menstruating rodent, Acomys cahirinus

BackgroundThe Egyptian spiny mouse (Acomys cahirinus) is the only known rodent to exhibit true, human-like menstruation and postpartum ovulation, and is an important new model for reproductive studies. Spiny mice do not produce a visible copulatory plug, and calculation of gestational age is therefore restricted by the need to use mated postpartum dams. The current inefficient method of monitoring until parturition to provide a subsequent estimate of gestational age increases study duration and costs. This study addressed this issue by comparing the mating behaviour of spiny mice across the menstrual cycle and proposes a more accurate method for staging and pairing animals that provides reliable estimates of gestational age. In experiment 1, mating behaviour was recorded overnight to collect data on mounting, intromission, and ejaculation (n = 5 pairs per stage) in spiny mice paired at menses and at early and late follicular and luteal phases of the menstrual cycle. In experiment 2, female spiny mice were paired at the follicular or luteal phases of the menstrual cycle to determine any effect on the pairing-birth interval (n = 10 pairs).ResultsWe report a broad mating window of ~ 3 days during the follicular phase and early luteal phase of spiny mice. Males displayed a discrete ‘foot twitch’ behaviour during intromission and a brief copulatory lock during ejaculation. Litters were delivered after 40–43 days if pairing occurred during the mating window, compared with 46–48 days for spiny mice paired in the late luteal phase. When pairing occurred during the late luteal phase or menses no mating activity was observed during the recording period.ConclusionThis study clearly demonstrates an effect of the menstrual cycle on mating behaviour and pregnancy in the spiny mouse and provides a reliable and more effective protocol for estimating gestational age without the need for postpartum dams.

Platelet-derived growth factor BB is reduced in endometrial endothelial cells of women with abnormal uterine bleeding–endometrial disorder

Research questionWhat is the expression pattern of platelet-derived growth factor BB (PDGF-BB), and its receptors, across the menstrual cycle in healthy control women and those with abnormal uterine bleeding–endometrial disorder (AUB-E)? DesignImmunohistochemical staining for PDGF-BB, platelet-derived growth factor receptor alpha (PDGFRα) and platelet-derived growth factor beta (PDGFRβ) was performed in control and AUB-E endometrium from the proliferative, early, mid- and late secretory phases of the menstrual cycle (n = 5 each group). Control proliferative phase endometrium was cultured in PDGF-BB (0, 10 ng/ml) and vascular maturation assessed (n = 3). Endothelial cell to vascular smooth muscle cell (VSMC) association was assessed after treatment with PDGF-BB (0, 1, 10 ng/ml). Secretion of angiogenic growth factors by endothelial cells or VSMC was determined. ResultsEndothelial cell immunoreactivity for PDGF-BB was reduced in the mid and late secretory phases in AUB-E (P = 0.008). PDGFRα was also reduced in mid secretory phase endothelial cells, proliferative and early secretory phase glandular epithelium in AUB-E (P = 0.008). PDGFRβ expression was not altered. Treatment of proliferative phase endometrium with PDGF-BB (10 ng/ml) reduced the percentage of vessels expressing contractile VSMC markers. PDGF-BB had no effect on angiogenic growth factor secretion by endothelial cells or VSMC in vitro and did not affect their association in an in-vitro endothelial cell–VSMC association assay. ConclusionsReduced endothelial cell expression of PDGF-BB in the AUB-E endometrium may contribute to the reduced vascular maturation previously observed in these women.

Form of Supplemental Selenium Affects the Expression of mRNA Transcripts Encoding Selenoproteins, and Proteins Regulating Cholesterol Uptake, in the Corpus Luteum of Grazing Beef Cows

Simple SummaryIn regions with selenium (Se) deficient soils, producers should supplement this mineral to the diet of forage-grazing cattle. Commercial supplements are typically formulated using inorganic forms of Se, but the organic forms are those naturally available in forages. We previously reported that circulating concentrations of progesterone are affected by the form of Se supplemented to cows. In this report, we aimed to determine (1) how the form of Se affects the expression of mRNA encoding selenoproteins in the corpus luteum, and (2) whether form of Se-induced increases in progesterone are the result of direct increases in the expression of steroidogenic transcripts. Cows were supplemented with the industry standard, an inorganic form of Se, or a 1:1 mix of organic and inorganic forms (MIX), with corpora lutea recovered on Day 7 of the estrous cycle. Transcripts encoding selenoproteins, as well as those regulating the uptake of cholesterol were affected by form of Se, but not those directly regulating steroidogenesis. We demonstrated that the expression of multiple selenoprotein mRNAs are affected by the form of Se, some of which are associated with intracellular antioxidant activity, and that the previously reported form of Se-induced increase in progesterone appears to be due to increased cholesterol availability by the corpus luteum, not by an increase in the expression of key steroidogenic enzymes.Selenium (Se)-deficient soils necessitate supplementation of this mineral to the diet of forage-grazing cattle. Functionally, Se is incorporated into selenoproteins, some of which function as important antioxidants. We have previously shown that the source of supplemental Se; inorganic (sodium selenite or sodium selenate; ISe), organic (selenomethionine or selenocysteine; OSe) or 1:1 mix of ISe and OSe (MIX), provided to Angus-cross cows affects concentrations of progesterone (P4) during the early luteal phase of the estrous cycle. In this study, we sought to investigate (1) the effect of form of Se on the expression of mRNA encoding selenoproteins in the corpus luteum (CL), and (2) whether this previously reported MIX-induced increase in P4 is the result of increased luteal expression of key steroidogenic transcripts. Following a Se depletion and repletion regimen, 3-year-old, non-lactating, Angus- cross cows were supplemented with either ISe as the industry standard, or MIX for at least 90 days, with the CL then retrieved on Day 7 post-estrus. Half of each CL was used for analysis of targeted mRNA transcripts and the remainder was dissociated for culture with select agonists. The expression of three selenoprotein transcripts and one selenoprotein P receptor was increased (p < 0.05), with an additional five transcripts tending to be increased (p < 0.10), in cows supplemented with MIX versus ISe. In cultures of luteal cells, hCG-induced increases in P4 (p < 0.05) were observed in CL obtained from ISe-supplemented cows. The abundance of steroidogenic transcripts in the CL was not affected by the form of Se, however, the abundance of mRNA encoding 2 key transcripts regulating cholesterol availability (Ldlr and Hsl) was increased (p < 0.05) in MIX-supplemented cows. Overall, the form of Se provided to cows is reported to affect the expression of mRNA encoding several selenoproteins in the CL, and that the form of Se-induced effects on luteal production of P4 appears to be the result of changes in cholesterol availability rather than a direct effect on the expression of steroidogenic enzymes within the CL.

Joint effect of particulate matter and cigarette smoke on women\u2019s sex hormones

BackgroundAlthough relationships between exposure to air pollution and reproductive health are broadly studied, mechanisms behind these phenomena are still unknown. The aim of the study was to assess whether exposure to particulate matter (PM10) and tobacco smoking have an impact on menstrual profiles of 17β-estradiol (E2) and progesterone (P) and the E2/P ratio.MethodsLevels of sex hormones were measured daily in saliva during the entire menstrual cycle among 132 healthy, urban women. Exposure to smoking (active or passive) was assessed by questionnaire, whilst exposure to PM10 with municipal monitoring data.ResultsDuring the early luteal phase, profiles of E2 were elevated among women with higher versus lower exposure to PM10 (p = 0.02, post-hoc tests). Among those who were exposed versus unexposed to tobacco smoking, the levels of mean E2 measured during the entire cycle were higher (p = 0.02). The difference in mean E2 levels between the group of joint exposure (i.e. to high PM10 and passive or active smoking) versus the reference group (low PM10, no smoking) was statistically significant at p = 0.03 (18.4 vs. 12.4 pmol/l, respectively). The E2/P ratios were higher among women with higher versus lower exposure to PM10 and this difference was seen only in the early luteal phase (p = 0.01, exploratory post-hoc tests).ConclusionsWe found that PM10 and tobacco smoking affect ovarian hormones independently and do not interact with each other. Both exposures appear to have estrogenic effects even though women's susceptibility to these effects differs across the menstrual cycle. We propose that the hormonal mechanisms are involved in observed relationships between air pollution and smoking with women’s reproductive health.

Role Of Ultrasound In Assessment Of Endometrial Thickness In Secretory Phase Of Infertile Females

Infertility in women is caused by a variety of hereditary and acquired uterine problems. Ultrasound has been found to be a good, quick, safe, and cost-effective way to assess the size, quantity, and location of abnormalities and to measure endometrial thickness in different phases of menstrual cycle.Objective: The main objective of this study wasto evaluate roleof ultrasound in assessment of endometrial thickness in infertile females of secretory phase. Methods: This is a descriptive study, including eighty two females age 24 to 46 years from University ultrasound clinic, Lahore after the approval of ethical committee of Allied health sciences, University of Lahore. Informed consent was taken before including the subjects in the study. Data was collected according to data collection sheets. Results: The results showed that primary infertility (54.9%) is more in females than secondary infertility (45.1%). There were 10(47.6%) females in primary infertility with 9-12mm endometrial thickness and 11(52.4%) in secondary infertility. 24(57.1%) females in primary infertility with 13-16mm endometrial thickness and 18(42.9%) in secondary infertility. 11(68.8%) females in primary infertility with 17-20mm endometrial thickness and 5(31.3%) in secondary infertility. 3(100%) females in secondary infertility with 21-24mm endometrial thickness.Conclusion: Female patients who initially taking infertility treatment had less endometrial thickness in proliferative phase; their endometrial thickness maybe increased in early secretory phase or at 21st day of their menstrual cycle but they had lost their chance to conceive the pregnancy in proliferative phase due to less endometrial thickness. The females should know the normal endometrial thickness in different phases of their menstrual cycle to conceive the pregnancy. This study shows ultrasound plays a vital role in detecting endometrial thickness in infertility treatment.

Menstrual cycle impacts lung structure measures derived from quantitative computed tomography.

Quantitative computed tomography (qCT) is being increasingly incorporated in research studies and clinical trials aimed at understanding lung disease risk, progression, exacerbations, and intervention response. Menstrual cycle-based changes in lung function are recognized; however, the impact on qCT measures is currently unknown. We hypothesize that the menstrual cycle impacts qCT-derived measures of lung structure in healthy women and that the degree of measurement change may be mitigated in subjects on cyclic hormonal birth control. Thirty-one non-smoking, healthy women with regular menstrual cycles (16 of which were on cyclic hormonal birth control) underwent pulmonary function testing and qCT imaging at both menses and early luteal phase time points. Data were evaluated to identify lung measurements which changed significantly across the two key time points and to compare degree of change across metrics for the sub-cohort with versus without birth control. The segmental airway measurements were larger and mean lung density was higher at menses compared to the early luteal phase. The sub-cohort with cyclic hormonal birth control did not have less evidence of measurement difference over the menstrual cycle compared to the sub-cohort without hormonal birth control. This study provides evidence that qCT-derived measures from the lung are impacted by the female menstrual cycle. This indicates studies seeking to use qCT as a more sensitive measure of cross-sectional differences or longitudinal changes in these derived lung measurements should consider acquiring data at a consistent time in the menstrual cycle for pre-menopausal women and warrants further exploration. • Lung measurements from chest computed tomography are used in multicenter studies exploring lung disease progression and treatment response. • The menstrual cycle impacts lung structure measurements. • Cyclic variability should be considered when evaluating longitudinal change with CT in menstruating women.

Cyclic activity of signal transduction pathways in fimbrial epithelium of the human fallopian tube.

The local environment of the fallopian tube represents the optimal conditions for reproductive processes. To maintain tissue homeostasis, signal transduction pathways are thought to play a pivotal role. Enhancing our understanding of functional signal transduction pathway activity is important to be able to clarify the role of aberrant signal transduction pathway activity leading to female subfertility and other tubal diseases. Therefore, in this study we investigate the influence of the hormonal cycle on the activity of key signal transduction pathways in the fimbrial epithelium of morphologically normal fallopian tubes. We included healthy pre- (n=17) and postmenopausal (n=8) patients who had surgical interventions for benign gynecologic conditions. Histologic sections of the fallopian tubes were reviewed by two pathologists and, for the premenopausal patients, hormone serum levels and sections of the endometrium were examined to determine the hormonal phase (early follicular [n=4], late follicular [n=3], early luteal [n=5], late luteal [n=5]). After laser capture microdissection, total mRNA was extracted from the fimbrial epithelium and real-time quantitative reverse transcription-PCR was performed to determine functional signal transduction pathway activity of the androgen receptor (AR), estrogen receptor (ER), phosphoinositide-3-kinase (PI3K), Hedgehog (HH), transforming growth factor-beta (TGF-β) and canonical wingless-type MMTV integration site (Wnt) pathways. The early luteal phase demonstrated high AR and ER pathway activity in comparison with the late luteal phase (p=0.016 and p=0.032, respectively) and low PI3K activity compared with the late follicular phase (p=0.036), whereas the late luteal phase showed low activity of HH and Wnt compared with the early follicular phase (both p=0.016). Signal transduction pathway activity in fimbrial epithelium from postmenopausal patients was most similar to the early follicular and/or late luteal phase with regard to the AR, ER and PI3K pathways. Wnt pathway activity in postmenopausal patients was comparable to the late follicular and early luteal phase. We observed no differences in HH and TGF-β pathway activity between pre- and postmenopausal samples. The cyclic changes in signal transduction pathway activity suggest a stage-specific function which may affect the morphology and physiology of the human fallopian tube. We demonstrated cyclic changes in activity of the AR, ER, PI3K, HH and Wnt pathways throughout the hormonal cycle.

Expression and role of resistin on steroid secretion in the porcine corpus luteum

Resistin plays an important role in adipogenesis, obesity, insulin resistance, and reproduction. Previous studies showed resistin action on ovarian follicular cells; however, whether resistin regulates steroid secretion in luteal cells is still unknown. Our aim was first to determine the expression of resistin and its potential receptors (tyrosine kinase-like orphan receptor 1 (ROR1) and toll-like receptor 4 (TLR4)) in the porcine corpus luteum (CL), regulation of its expression, effect on kinases phosphorylation, and luteal steroidogenesis. Our results showed that the expression of resistin and its receptors was dependent on the luteal phase and this was higher at the mRNA level in the late compared with the early and middle luteal phase. At the opposite, resistin protein expression was higher in the middle and late compared with the early luteal phase, while ROR1 and TLR4 expression was highest in the early luteal phase. Additionally, we observed cytoplasmic localisation of resistin, ROR1, and TLR4 in small and large luteal cells. We found that luteinising hormone, progesterone (P4), insulin, and insulin-like growth factor 1 regulated the protein level of resistin, ROR1, and TLR4. Resistin decreased P4 and increased oestradiol (E2) secretion via changes in steroidogenic enzymes expression and via the activation of protein kinase A (PKA) and mitogen-activated protein kinase (MAP3/1), increased the expression of receptors LHCGR and ESR2 and decreased the expression of PGR. Moreover, resistin decreased PKA phosphorylation and enhanced MAP3/1 phosphorylation. Taken together, resistin could act directly on steroid synthesis and serve as an important factor in in vivo luteal cell function.

Steroid stress hormone changes throughout the menstrual cycle: A rise in evening aldosterone concentration in early luteal phase precedes the symptoms of premenstrual syndrome.

There is still a lack of consistent evidence on dysregulation of steroid stress hormones cortisol and aldosterone in premenstrual syndrome (PMS). We aimed to test the hypothesis that, in healthy women, salivary aldosterone concentrations are higher in those with PMS compared to controls, particularly during the luteal phase of the menstrual cycle. In total, 99 female subjects (49 women with and 50 women without PMS) participated in a prospective non-interventional case-control study. Saliva sampling was performed in the follicular (day 8), early luteal (day 20) and late luteal phase (2days before expected onset of bleeding) of the menstrual cycle in the morning and the evening. The results confirmed the hypothesis that salivary aldosterone concentrations are higher in women with PMS during the early luteal phase compared to controls (p<.01) in the evening. Early luteal phase aldosterone concentrations positively correlated with the frequency of premenstrual symptoms. Women with PMS exhibited a flatter morning to evening aldosterone slope compared to controls (p<.05). Morning and evening salivary cortisol concentrations were unchanged throughout the menstrual cycle in both groups of women. In conclusion, evening salivary aldosterone, but not cortisol concentrations, are increased in women with PMS during the early luteal phase compared to controls. Cortisol does not appear to be involved in the mechanisms contributing to the course of PMS. High evening salivary aldosterone in the early luteal phase may represent an important risk factor and could be of predictive value for the occurrence of premenstrual symptoms.

Effects of cortisol on prostaglandin F2α secretion and expression of genes involved in the arachidonic acid metabolic pathway in equine endometrium - In vitro study

Glucocorticoids (GCs) are known to play an important role in maintaining basal and stress-related homeostasis by interacting with endocrine mediators and prostaglandins (PGs). Although a growing body of evidence shows that GCs exert their regulatory action at a multitude of sites in the reproductive axis through corticosteroid receptors, little is known about the direct role of cortisol, an active form of GCs, in the equine endometrium. Thus, the study aimed to determine the effect of cortisol on PGF2α synthesis in the endometrial tissue and cells in vitro. In Exp.1, the immunolocalization and the expression of the glucocorticoid receptor (GCR) in the endometrium throughout the estrous cycle were established. In Exp. 2 and 3, the effects of cortisol on PGF2α secretion and transcripts associated with the arachidonic acid (AA) cascade in endometrial tissues, and cells were defined. Endometrial tissues obtained from the early, mid, and late luteal phases and the follicular phase of the estrous cycle were exposed to cortisol (100, 200, and 400 nM) for 24 h. Endometrial epithelial and stromal cells (early phase of estrous cycle) were exposed to cortisol (100 nM) for 24 h. Then, PGF2α secretion and transcripts associated with the AA cascade (PLA2G2A, PLA2G4A, PTGS2, and PGFS) were assessed. GCR was expressed in the cytoplasm and the nucleus in the luminal and glandular epithelium as well as in the stroma. Endometrial GCR protein abundance was up-regulated at the late luteal phase compared to the mid-luteal phase of the estrous cycle. Cortisol dose-dependently decreased PGF2α secretion, PLA2G2A and PLA2G4A transcripts in endometrial tissues. Additionally, cortisol treatment decreased PGF2α secretion from endometrial epithelial and stromal cells. Moreover, it affected PLA2G2A, PLA2G4A, and PTGS2 transcripts in endometrial stromal cells. These findings suggest that cortisol suppresses the synthesis of PGF2α by affecting the AA cascade in the equine endometrium during the estrous cycle.

P–681 Will the hCG trigger dose used for final oocyte maturation in IVF impact endogenous progesterone during the luteal phase? - A randomized controlled trial

Abstract Study question Is there an association between the hCG dose used for ovulation trigger and the endogenous progesterone production during the luteal phase? Summary answer Increased hCG dosing significantly increased the endogenous progesterone level during the luteal phase. What is known already During the luteal phase of an IVF treatment, the endogenous progesterone (P4) production is negatively impacted due to reduced circulating endogenous LH, caused by negative feed-back of elevated steroids; thus, luteal phase support (LPS) with exogenous P4 remains mandatory in IVF. Apart from inducing final oocyte maturation, the gold standard HCG trigger also functions as an early LPS, boosting P4 production by the corpora lutea (CL). P4 plays a pivotal role for embryo implantation and pregnancy, and an optimal P4 level around peri-implantation seems to be essential for the reproductive outcomes of fresh and frozen/thaw embryo transfer cycles. Study design, size, duration A randomized controlled 4-arm study, including a total of 127 IVF patients, enrolled from January 2015 until September 2019 at the Fertility Clinic, Odense University Hospital, Denmark. Participants/materials, setting, methods IVF patients with ≤ 11 follicles ≥ 12 mm were randomized to four groups. Groups 1–3 were triggered with: 5.000 IU, 6.500 IU or 10.000 IU, hCG, respectively, receiving a LPS consisting of 17-α-hydroxy-progesterone (17α OH P4) to distinguish the endogenous P4 from the exogenous supplementation. Group 4 (control) was randomized to a 6.500 IU hCG trigger and standard LPS. A total of eight blood samples were drawn during the early luteal phase. Main results and the role of chance A total of 94 patients completed the study: 21, 22, 25 and 26 patients in each group, respectively. Baseline characteristics were similar, except for the endogenous LH level and cycle lengths. There were no significant differences between groups regarding ovarian stimulation, number of oocytes and embryos. The median number of follicles ≥ 12mm on the day of trigger was 8.5, resulting in 6.6 oocytes being retrieved. Significant differences in P4 levels were seen at OPU+8 (p &amp;lt; 0.001), OPU+10 (p &amp;lt; 0.001) and OPU+14 (p &amp;lt; 0.001), with positive correlations between P4 level and hCG dose. Groups compared individually showed significant difference in P4 between low and high trigger dose at OPU+4 group 1 and 3 (p = 0.037) and OPU+8 group 1 and 3 (p = 0.007) and between all the three groups around implantation at OPU+6 group 1 and 2 (p = 0.011), group 2 and 3 (p = 0.042) and group 1 and 3 (p &amp;lt; 0.001). Higher P4 levels around implantation were related to follicle count and to pregnancy. After logistic regression analyses there were still significant individual differences between the groups. Limitations, reasons for caution Although patients were randomized and strict inclusion and exclusion criteria were used, the RCT was un-blinded, including a relatively small number of patients. Moreover, for dosing purposes urinary hCG as well as recombinant hCG was used and pharmacokinetics differ. Finally, the P4 level could be influenced by circadian fluctuations. Wider implications of the findings: This is the first study to explore dose-responses in circulating P4 after hCG trigger in IVF patients. Increasing the hCG trigger dose increased the endogenous P4 around peri-implantation. Personalizing the hCG trigger dose could be a key point to secure the most optimal P4 mid-luteal phase P4 level. Trial registration number Eudract 2013–003304–39

P-681 Will the hCG trigger dose used for final oocyte maturation in IVF impact endogenous progesterone during the luteal phase? - A randomized controlled trial

Abstract Study question Is there an association between the hCG dose used for ovulation trigger and the endogenous progesterone production during the luteal phase? Summary answer Increased hCG dosing significantly increased the endogenous progesterone level during the luteal phase. What is known already During the luteal phase of an IVF treatment, the endogenous progesterone (P4) production is negatively impacted due to reduced circulating endogenous LH, caused by negative feed-back of elevated steroids; thus, luteal phase support (LPS) with exogenous P4 remains mandatory in IVF. Apart from inducing final oocyte maturation, the gold standard HCG trigger also functions as an early LPS, boosting P4 production by the corpora lutea (CL). P4 plays a pivotal role for embryo implantation and pregnancy, and an optimal P4 level around peri-implantation seems to be essential for the reproductive outcomes of fresh and frozen/thaw embryo transfer cycles. Study design, size, duration A randomized controlled 4-arm study, including a total of 127 IVF patients, enrolled from January 2015 until September 2019 at the Fertility Clinic, Odense University Hospital, Denmark. Participants/materials, setting, methods IVF patients with ≤ 11 follicles ≥ 12 mm were randomized to four groups. Groups 1-3 were triggered with: 5.000 IU, 6.500 IU or 10.000 IU, hCG, respectively, receiving a LPS consisting of 17-α-hydroxy-progesterone (17α OH P4) to distinguish the endogenous P4 from the exogenous supplementation. Group 4 (control) was randomized to a 6.500 IU hCG trigger and standard LPS. A total of eight blood samples were drawn during the early luteal phase. Main results and the role of chance A total of 94 patients completed the study: 21, 22, 25 and 26 patients in each group, respectively. Baseline characteristics were similar, except for the endogenous LH level and cycle lengths. There were no significant differences between groups regarding ovarian stimulation, number of oocytes and embryos. The median number of follicles ≥ 12mm on the day of trigger was 8.5, resulting in 6.6 oocytes being retrieved. Significant differences in P4 levels were seen at OPU+8 (p &amp;lt; 0.001), OPU+10 (p &amp;lt; 0.001) and OPU+14 (p &amp;lt; 0.001), with positive correlations between P4 level and hCG dose. Groups compared individually showed significant difference in P4 between low and high trigger dose at OPU+4 group 1 and 3 (p = 0.037) and OPU+8 group 1 and 3 (p = 0.007) and between all the three groups around implantation at OPU+6 group 1 and 2 (p = 0.011), group 2 and 3 (p = 0.042) and group 1 and 3 (p &amp;lt; 0.001). Higher P4 levels around implantation were related to follicle count and to pregnancy. After logistic regression analyses there were still significant individual differences between the groups. Limitations, reasons for caution Although patients were randomized and strict inclusion and exclusion criteria were used, the RCT was un-blinded, including a relatively small number of patients. Moreover, for dosing purposes urinary hCG as well as recombinant hCG was used and pharmacokinetics differ. Finally, the P4 level could be influenced by circadian fluctuations. Wider implications of the findings This is the first study to explore dose-responses in circulating P4 after hCG trigger in IVF patients. Increasing the hCG trigger dose increased the endogenous P4 around peri-implantation. Personalizing the hCG trigger dose could be a key point to secure the most optimal P4 mid-luteal phase P4 level. Trial registration number Eudract 2013-003304-39

P-631 Embryo euploidy rates following follicular or luteal start ovarian stimulation. A prospective study with repeated ovarian stimulation ovarian stimulation cycles

Abstract Study question Is there any difference in embryo euploidy rates following luteal phase phase (LS) and follicular phase (FS) start ovarian stimulation. Summary answer The number of euploid blastocysts and embryo euploidy rate are comparable when comparing FS and LS. What is known already Random start ovarian stimulation (starting at any time of the cycle) has been traditionally used in women undergoing urgent fertility preservation for medical reason. Although there is accumulating evidence that in infertile women, LS can result in equivalent number of oocytes and embryos as compared with FS, no study has evaluated the effect of luteal phase start ovarian stimulation on embryo euploidy rates. The current study is the first prospective study designed to evaluate embryo euploidy rates in donors undergoing two identical consecutive ovarian stimulation protocols within a period of 6 months starting either in the (FS), or (LS). Study design, size, duration In a prospective study, conducted between May 2018 and January 2020, 40 oocyte donors underwent two consecutive ovarian stimulation protocols within a period of 6 months with an identical fixed GnRH antagonist protocol starting either in the early follicular (FS), or and luteal menstrual cycle phase (LS). Participants/materials, setting, methods All participants underwent two identical consecutive ovarian stimulation cycles with 150μg corifollitropin alfa followed by 200 IU rFSH in a fixed GnRH antagonist protocol either in the FS or LS. Six MII oocytes from the same oocyte donor, from each stimulation cycle, were allocated to the recipients and were inseminated with the same sperm sample (recipients partner sperm or donor sperm). Embryos were cultivated to blastocyst stage followed by preimplantation genetic testing for aneuploidies (PGT-A). Main results and the role of chance When comparing FP with LP, the duration of ovarian stimulation was significantly shorter (9.68± 2.09 vs 10.93± 1.55 days), 95% CI [-1.95; -0.55] and a higher total additional dose of daily recFSH was significantly lower (526.14± 338.94 IU vs 726.14± 366.27), 95% CI [-315,12; -84,88] when CPT was administered in the luteal phase. . There were no differences in the hormone values on the triggering day (Estradiol 2137.61±1198.25 pg/ml vs 2362.96±1472.89); 95% CI [-1160.45;709.76]. Overall no differences were observed in the number of oocytes (24.84± 11.200 vs 24.27± 9.08); 95% CI[-2,61; 3.75] and MII oocytes (21.41±10.19 vs 21.59± 8.81), 95%CI [-2.72; 2.35] retrieved between FP and LP cycles in the oocytes donors. Following oocyte allocation and fertilization to the recipients, a total of 245 blastocysts were biopsied (blastocyst formation rate 245/408, 60.05%), 117 in FP group and 128 in LP group. The overall blastocyst euploidy rate was 59.18% . There were no differences in the number of euploid embryos between FS (1.59±1.32) and LS (1.70±1.29), mean difference 0.11, 95%CI [-0.65; 0.46]. Finally, there were no differences in the percentage of euploid embryos per oocytes inseminated between FS [70/287 (24.4%)] and LP [75/278 (24.7%), mean difference -0.027, 95%CI [-0.11; 0.06]. Limitations, reasons for caution The study was performed in oocyte derived from potentially fertile young oocyte donors thus caution is needed when extrapolating the results in oocytes derived from infertile women of older age. Wider implications of the findings Luteal phase stimulation does not alter embryo euploidy status as compared with follicular phase stimulation and thus it appears that it can be safely used not only in cases of urgent medical fertility preservation but also in patients undergoing ovarian stimulation for IVF/ICSI. Trial registration number Clinical Trials Gov (NCT03555942).