Incidence Of Early Pregnancy Loss Research Articles

A randomized, controlled clinical study of low-molecular-weight heparin improving pregnancy outcomes in PCOS women undergoing IVF: study protocol

BackgroundPolycystic ovary syndrome (PCOS), an incidence of 10–15% in women of reproductive age, shows sex hormone disorders, luteal insufficiency, and the tendency of placental villus space thrombus. The incidence of early pregnancy loss in women with PCOS is three to eight times higher than that in non-PCOS women. PCOS women were reported in a pre-thrombotic state, which was manifested by accelerated thrombin production, increased PAI-1 activity, and fibrinogen. Other research also found an over-activated state of women with PCOS in immune system. Therefore, changing the prethrombotic state of PCOS through anticoagulation may be a new way to improve the adverse pregnancy outcome of PCOS. Low-molecular-weight heparin (LMWH) is the most common used anticoagulant drug in pregnancy, and it also was proposed for the prevention of recurrent abortion, although the application of LMWH in PCOS population during early pregnancy has not been reported. The objective of this study is to investigate the effect of LMWH on pregnancy outcomes after invitro fertilization-frozen embryo transfer (IVF-FET) in patients with polycystic ovary syndrome.MethodsA total of 356 PCOS women aged between 20 and 38 years which prepared for IVF followed with FET will be enrolled in the study. The patients, from four different hospitals stratified by age and body mass index (BMI), will be randomly divided into the study group who will be treated with LMWH started on the day of progesterone transformation (hormone therapy) during FET cycle and the control group without additional medicine. Serum or urine hCG test will be given 14 days after embryo transfer to confirm biochemical pregnancy. If pregnancy is positive, LMWH+ hormone therapy/hormone therapy will be continued for another 2 weeks. Transvaginal ultrasonography will be performed 14 days later to confirm intrauterine pregnancy. The primary outcome is the ongoing pregnancy, which is defined as intrauterine live fetus with ultrasound after 12 weeks of gestation.DiscussionThis is the first study protocol to investigate the efficacy of LMWH as an adjuvant drug for IVF-FET outcomes in PCOS women, by comparing differences in ongoing pregnancy rate, clinical pregnancy rate, live birth rate, and early pregnancy loss rate between LMWH group and the control group.Trial registration ClinicalTrials.gov ChiCTR2000036527. Registered on August 24, 2020

Genomic analyses of wild argali, domestic sheep, and their hybrids provide insights into chromosome evolution, phenotypic variation, and germplasm innovation.

Understanding the genetic mechanisms of phenotypic variation in hybrids between domestic animals and their wild relatives may aid germplasm innovation. Here, we report the high-quality genome assemblies of a male Pamir argali (O. ammon polii, 2n = 56), a female Tibetan sheep (O. aries, 2n = 54), and a male hybrid of Pamir argali and domestic sheep, and the high-throughput sequencing of 425 ovine animals, including the hybrids of argali and domestic sheep. We detected genomic synteny between Chromosome 2 of sheep and two acrocentric chromosomes of argali. We revealed consistent satellite repeats around the chromosome breakpoints, which could have resulted in chromosome fusion. We observed many more hybrids with karyotype 2n = 54 than with 2n = 55, which could be explained by the selfish centromeres, the possible decreased rate of normal/balanced sperm, and the increased incidence of early pregnancy loss in the aneuploid ewes or rams. We identified genes and variants associated with important morphological and production traits (e.g., body weight, cannon circumference, hip height, and tail length) that show significant variations. We revealed a strong selective signature at the mutation (c.334C > A, p.G112W) in TBXT and confirmed its association with tail length among sheep populations of wide geographic and genetic origins. We produced an intercross population of 110 F2 offspring with varied number of vertebrae and validated the causal mutation by whole-genome association analysis. We verified its function using CRISPR-Cas9 genome editing. Our results provide insights into chromosomal speciation and phenotypic evolution and a foundation of genetic variants for the breeding of sheep and other animals.

O-272 Letrozole-induced endometrial preparation improved pregnancy outcomes after frozen blastocyst transfer compared to the natural cycle

Abstract Study question Does letrozole-induced endometrial preparation affect pregnancy outcomes, perinatal outcomes, and congenital anomalies after single vitrified-warmed blastocyst transfers (SVBTs) when compared with the natural cycle? Summary answer Letrozole-induced endometrial preparation improved live birth rate without any adverse effects on perinatal outcomes and congenital anomalies after SVBTs. What is known already Letrozole treatment is considered an effective option in endometrial preparation for frozen embryo transfers in patients with ovulation disorders or irregular menstruation; however, the effectiveness of letrozole-induced endometrial preparation is still unclear in ovulatory patients. Furthermore, there is no comparative study reporting on pregnancy complications and congenital anomalies after frozen embryo transfers comparing natural and letrozole-assisted cycles. Study design, size, duration This retrospective study, at a major academic fertility centre, analysed a total of 14,611 clinical records of women who underwent SVBTs comprising both natural and letrozole-assisted cycles between July 2015 and June 2020. The cycle characteristics, pregnancy outcomes (clinical pregnancy, ongoing pregnancy, and live birth), and the incidence of pregnancy complications and congenital anomalies were statistically compared between the natural and letrozole groups. Participants/materials, setting, methods The study reviewed ovulatory patients who underwent their first SVBT during the study period. Some patients took letrozole during the early proliferative phase to promote follicular development and maturation (letrozole group). Ovulation was triggered by GnRH agonist and SVBTs were performed on day five after ovulation. Propensity score matching was performed to reduce any bias from patient characteristics. Multivariate logistic analysis was performed to evaluate the effects of letrozole administration on pregnancy and perinatal outcomes. Main results and the role of chance After propensity score matching, the characteristics of patients and transferred blastocysts were comparable between groups. The serum progesterone level was also significantly increased in the letrozole group (P < 0.0001). Although no difference was observed regarding implantation rate between groups, the rates of clinical pregnancy, ongoing pregnancy, and delivery in the letrozole group were all significantly higher than that in the natural group (P = 0.0273, P = 0.0162, P = 0.0479, respectively). The incidence of early pregnancy loss, miscarriage, and stillbirth were comparable between groups. Multivariate logistic regression analysis also demonstrated that the administration of letrozole during an SVBT cycle significantly improved the live birth rate (AOR, 1.160; P = 0.0355). The incidence of pregnancy complications was comparable between groups. The caesarean section rate was significantly lower in the letrozole group than that in the natural group (P = 0.0464). Gestational age, birth length, birth weight, and infant sex, as well as the incidence of pregnancy complications and birth defects, were statistically comparable between groups. Furthermore, multivariate logistic regression analysis showed that perinatal outcomes were not affected by letrozole-induced endometrial preparation, although the incidence of caesarean section was decreased in the letrozole group (AOR, 0.788; P = 0.0355). Limitations, reasons for caution Our findings are not compared with reported incidences of pregnancy complications and congenital anomalies in natural pregnancy. Furthermore, the study was retrospective in nature, and further multicentre studies are required to ascertain the generalisability of these findings for other clinics with different protocols and/or patient demographics. Wider implications of the findings Letrozole administration both extended the proliferative phase and increased luteal function, resulting in an improvement of live birth rates without any adverse effects. Therefore, letrozole-induced endometrial preparation might be a safe and more effective strategy for patients with shortened proliferative phase or insufficient luteal function. Trial registration number not applicable

Early pregnancy loss incidence in high-income settings: a protocol for a systematic review and meta-analysis

BackgroundEarly pregnancy loss (unintended pregnancy loss before 20 completed weeks of gestation) is a common adverse pregnancy outcome, with previous evidence reporting incidence ranging from 10 to 30% of detected pregnancies. The objective of this systematic review and meta-analysis is to determine the incidence and range of early pregnancy loss in contemporary pregnant populations based on studies with good internal and external validity. Findings may be useful for clinical counseling in pre-conception and family planning settings and for people who experience early pregnancy loss.MethodsWe will search MEDLINE, EMBASE, and CINAHL databases using combinations of medical subject headings and keywords. Peer-reviewed, full-text original research articles that meet the following criteria will be included: (1) human study; (2) study designs: controlled clinical trials or observational studies with at least 100 pregnancies in the denominator, or systematic reviews of studies using these designs; (3) conducted in high-income countries; (4) reporting early pregnancy loss incidence, defined as unintended early pregnancy loss occurring prior to 20 weeks’ gestation expressed as the number of losses among all pregnancies in the study period; (5) among a contemporary (1990 or later) general population of pregnancies; and (6) published between January 1, 1990, and August 31, 2021. We will assess the quality of included studies according to the United States Preventive Services Task Force Criteria for Assessing Internal and External Validity of Individual Studies. If appropriate, based on methodological comparability across included studies, we will conduct meta-analyses using random effects models to estimate the pooled incidence of early pregnancy loss among all studies with both good internal and external validity, with meta-analyses stratified by study design type (survey-based or self-reported and medical record-based), by induced abortion restrictions (restricted vs. unrestricted), and by gestational age (first trimester only vs. all gestational ages before 20 weeks).DiscussionThis systematic review will synthesize existing evidence to calculate a current estimate of early pregnancy loss incidence and variability in reported incidence estimates in high-income settings. The findings of this review may inform updates to clinical counseling in pre-conception and family planning settings, as well as for patients experiencing early pregnancy loss.Systematic review registrationWe have registered this review with the International Prospective Register of Systematic Reviews (PROSPERO #226267).

Excessive endoplasmic reticulum stress drives aberrant mouse trophoblast differentiation and placental development leading to pregnancy loss.

Endoplasmic reticulum (ER) stress promotes placental dysmorphogenesis and is associated with poor pregnancy outcomes. We show that unfolded protein response signalling pathways located in the ER drive differentiation of mouse trophoblast stem cells into trophoblast subtypes involved in development of the placental labyrinth zone and trophoblast invasion. In a mouse model of chronic ER stress (Eif2s1tm1RjK ), higher ER stress in homozygous blastocysts is accompanied by reduced trophectoderm cell number and developmental delay and also is associated with an increased incidence of early pregnancy loss. Administration of the chemical chaperone, tauroursodeoxycholic acid, to Eif2s1+/tm1RjK heterozygous females during pregnancy alleviated ER stress in the mutant placenta, restored normal trophoblast populations and reduced the frequency of early pregnancy loss. Our results suggest that alleviation of intrauterine ER stress could provide a potential therapeutic target to improve pregnancy outcome in women with pre-gestational metabolic or gynaecological conditions. Women with pre-gestational health conditions (e.g. obesity, diabetes) or gynaecological problems (e.g. endometriosis) are at increased risk of adverse pregnancy outcomes including miscarriage, pre-eclampsia and fetal growth restriction. Increasing evidence suggests that unfavourable intrauterine conditions leading to poor implantation and/or defective placentation are a possible causative factor. The endoplasmic reticulum (ER) unfolded protein response (UPRER ) signalling pathways are a convergence point of various physiological stress stimuli that can be triggered by an unfavourable intrauterine environment. Therefore, we explored the impact of ER stress on mouse trophoblast differentiation in vitro, mouse blastocyst formation and early placenta development in the Eif2s1tm1RjK mutant mouse model of chronic ER stress. Chemically-manipulated ER stress or activation of UPRER pathways in a mouse trophoblast stem cell line promoted lineage-specific differentiation. Co-treatment with specific UPRER pathway inhibitors rescued this effect. Although the inner cell mass was unaffected, the trophectoderm of homozygous Eif2s1tm1RjK blastocysts exhibited ER stress associated with a reduced cell number. Furthermore, one-third of Eif2s1tm1RjK homozygous blastocysts exhibited severe developmental defects. We have previously reported a reduced trophoblast population and premature trophoblast differentiation in Eif2s1tm1RjK homozygous placentas at mid-gestation. Here, we demonstrate that treatment of Eif2s1+/tm1RjK heterozygous pregnant females with the chemical chaperone tauroursodeoxycholic acid alleviated ER stress, restored the trophoblast population and reduced the frequency of embryonic lethality. Our data suggest that therapeutic targeting of ER stress may improve pregnancy outcome in women with pre-gestational metabolic or gynaecological conditions.

Pregnancy and parturition in dromedary camels II. Incidence, timing and factors affecting early pregnancy loss (EPL) and the outcome of twin pregnancies

In this paper, we described the incidence of early pregnancy loss (EPL) both after natural mating and embryo transfer, evaluated risk factors, and summarized the outcome of twin pregnancies throughout gestation in dromedaries under reproductive care. Data were collected over seven breeding seasons at the world's largest camel dairy farm (study 1). In addition, we determined the timing of EPL and monitored serum progesterone (P4) concentration between Days 13–70 of gestation during one breeding season (study 2). In the first study, out of 2970 pregnancies, 507 cases (17.1%) of EPL were diagnosed with transrectal ultrasonography. The rate of EPL after natural mating and embryo transfer was 16.1% (n = 422 out of 2616) and 24.0% (n = 85 out of 354), respectively. Twin pregnancies were detected in 215 cases (7.2% of all gestations), and 57 of those (26.5%) underwent complete EPL. Almost half of the early losses (n = 243; 47.9%) occurred before 30 d of gestation. Another 43.2% (n = 219) of EPL was diagnosed during the next month, and 8.9% (n = 45) occurred after 60 d of gestation. Multivariable mixed effects logistic regression models revealed that the breeding season (year) and twin pregnancy were the most important exposure variables affecting the rate of EPL (P < 0.001). The effect of some male camels was also demonstrated while other factors, such as type of breeding, age category, month of mating, breed/ecotype and reproductive history did not prove to have a significant influence. In the second study, the overall rate of EPL was 24.5% (n = 34 of 139). There was no difference in the incidence of EPL between ET recipient (24.2%, n = 23 of 95) and mated (25%, n = 11 of 44) camels. Weekly rate of EPL ranged from 0.9% to 4.8% with a decreasing tendency, and approx. 41% of the animals (n = 14 of 34) had some ultrasonographic signs of impending EPL 1 week before the final diagnosis. Mean serum P4 concentration in camels with subsequent EPL was 5.3 ± 0.1 ng/ml compared to 5.6 ± 0.04 ng/ml in normal pregnant dromedaries. Day of gestation and future EPL influenced serum P4 levels (P < 0.001) with an interaction between the two fixed factors (P < 0.05). At the time of the final diagnosis of EPL, mean serum P4 concentration was 2.8 ± 0.44 ng/ml. Although twinning had an unfavorable prognosis with a total pregnancy loss of 36.7%, it was not entirely detrimental for the final outcome of gestation as two-thirds of twin pregnancies (n = 136 out of 212) resulted in the birth of a live calf.

SCM-198 ameliorates endometrial inflammation via suppressing the LPS-JNK-cJUN/cFOS-TLR4-NF-κB pathway.

Endometritis is an inflammatory disease of the endometrium, which is responsible for endometrial dysfunction, decidualization failure, and increased incidence of early pregnancy loss. SCM-198, a synthetic form of leonurine, is well known to possess anti-inflammatory effects. SCM-198 has been reported to display beneficial effects on endometritis. However, the specific mechanisms of SCM-198 in preventing endometritis remain unknown. In this study, we focused on the molecular mechanism of SCM-198 in inhibiting endometritis. The anti-inflammatory effects and the related signaling pathways of SCM-198 were studied in vitro using human endometrial stromal cells (hESCs). Reverse transcriptase-polymerase chain reaction and western blot analysis results demonstrated that SCM-198 markedly inhibited lipopolysaccharide (LPS)-induced endometrial inflammatory response by suppressing the LPS-JNK-cJUN/cFOS-TLR4-NF-κB pathway. The preventive and therapeutic effects of SCM-198 on endometrial inflammation were explored by using a mouse model of LPS-induced endometritis. SCM-198 produced essentially the same effects when administered either post-treatment (after LPS) or pre-treatment (before LPS) via vaginal or intraperitoneal administration. In vivo results indicated that SCM-198 is a potential effective drug for the treatment of endometritis.

Pathological Examination of the Late Embryonic Heart Using the Same 4-chamber and 3-vessel Planes Used in Fetal Echocardiography.

The incidence of early pregnancy loss widely varies according to age, being considerably higher in older women. Severe congenital malformations play an important role in pregnancy loss, having a high risk of recurrence. Congenital heart defects are the most common congenital abnormalities, thus the diagnosis of such malformations in aborted embryos is important for establishing both a possible cause for pregnancy loss and for correctly counseling the parents. Pathologic examination of the heart that is only a few millimeters in size, is very challenging. A pathologic examination protocol using transverse microscopic sections at the level of the 4-chamber and 3-vessel planes is proposed for heart evaluation. Two 9-10 gestational weeks embryos were microscopically examined using transverse slides of the thorax. The 4-chamber and 3-vessel slides were analyzed and compared to 11-13 weeks ultrasound images of the 4-chamber and 3-vessel views from 10 cases. The pathologic examination provided a detailed view of the ventricles, atria and great vessels, sometime surpassing even the ultrasound examination that was performed at a later gestational age. We consider our proposed pathologic examination protocol feasible for evaluating normal heart structures and ruling out severe congenital heart disease.

Maternal Vitamin D level and Early Pregnancy Loss A Nested Case Control Study

Objective: To assess the role of vitamin D deficiency and early pregnancy loss.Patient and Method: A nested case control study conducted in outpatient antenatal care clinic under supervision of Ain Shams University Maternity hospital from the period of March 2020 to June2020. Pregnant ladies in the first trimester were screened for eligibility criteria.Blood samples were taken from the participants at thetime of presentation. All participants were followed till the end of thefirst trimester to report cases of miscarriage. Vitamin d was assessedfor the 40 women who suffered from miscarriage (cases) and for 40selected controls.The primary outcome was the relation between vitamin d level and early pregnancy loss. Secondary outcome was therelation between vitamin d deficiency and obesity.Results: Our findings showed that the miscarriage groupwas significantly olderthan control group as p (42.5%) had 25(OH)D deficiency while (40.0%)&(17.5%) of caseshad either 25(OH)D insufficiencyor sufficiency which significantlydifferent than control group (p=0.049).25(OH)D ≤24.5 (ng/mL) wasa significant factor that increased the likelihood of first-trimestricmiscarriage with sensitivity 80%.No significant differences according to BMI grades regarding 25(OH)D grades.Conclusion: Vitamin d deficiency is one of modifiable risk factors forfirst trimestric abortion. . Preconceptional vitamin D supplementationis an easy method for decreasing incidence of early pregnancy loss.

Embryo-maternal communication during the establishment of equine pregnancy

Over the last 30 years, improvements in veterinary management have resulted in higher per cycle pregnancy rates in commercial horse breeding programmes. However, the incidence of early pregnancy loss has not diminished and ~ 10% of pregnancies detected on day 15 fail to survive beyond day 42 of pregnancy. To develop strategies to combat pregnancy loss, it is first necessary to understand why these pregnancies fail. In this respect, a sizeable proportion probably results from intrinsic embryonic abnormalities that cannot be remedied; another significant subset is, however, likely to result from an ‘inadequately receptive’ uterus or failure of the uterus to physiologically adapt to the developing conceptus’ changing needs, as a result of inadequate embryo-maternal communication. Communication between embryo and dam starts during the oviductal period, as evidenced by selective transport of viable embryos into the uterus on day 6 postovulation, whereas unfertilized oocytes remain in the oviduct. Conversely, retarded development exhibited by embryos produced in vitro demonstrates the importance of the oviductal environment to the early embryo. Once in the uterus, the embryo must steer maternal physiology to ensure adequate nutrient provision and prepare the endometrium for implantation. This communication includes ‘maternal recognition of pregnancy’ signalling to extend the lifespan of the primary corpus luteum and ensure continued secretion of progesterone. Although the identity of the embryonic pregnancy recognition signal remains unknown, our understanding of how endometrial function is altered to avert luteolytic prostaglandin F2α release and become receptive to implantation has improved.

Early pregnancy loss in patients with polycystic ovary syndrome after IVM versus standard ovarian stimulation for IVF/ICSI.

Is the incidence of early pregnancy loss (EPL) in patients with polycystic ovary syndrome (PCOS) higher after IVM of oocytes than after ovarian stimulation (OS) for IVF/ICSI? Women with PCOS who are pregnant after fresh embryo transfer have a higher probability of EPL following IVM, but after frozen embryo transfer (FET), no significant difference in the incidence of EPL was observed following IVM compared to OS. There is conflicting evidence in the current literature with regard to the risk of EPL after IVM of oocytes when compared with OS. Because of the limited sample size in previous studies, the use of different IVM systems and the possible bias introduced by patient characteristics and treatment type, firm conclusions cannot be drawn. This was a retrospective cohort study evaluating 800 women, with a diagnosis of infertility and PCOS as defined by Rotterdam criteria, who had a first positive pregnancy test after fresh or FET following IVM or OS between January 2010 and December 2017 in a tertiary care academic medical centre. Pregnancies after non-hCG triggered IVM following a short course of highly purified human menopausal gonadotropin were compared with those after conventional OS. The primary outcome was EPL, defined as a spontaneous pregnancy loss before 10 weeks of gestation. In total, 329 patients with a positive pregnancy test after IVM and 471 patients with a positive pregnancy test after OS were included. Women who were pregnant after IVM were younger (28.6 ± 3.4 years vs 29.3 ± 3.6 years, P = 0.005) and had higher serum anti-Mullerian hormone levels (11.5 ± 8.1 ng/ml vs 7.2 ± 4.1 ng/ml, P < 0.001) compared to those who were pregnant after OS. The distribution of PCOS phenotypes was significantly different among women in the IVM group compared to those in the OS group and women who were pregnant after OS had previously suffered EPL more often (28% vs 17.6%, P = 0.003). EPL was significantly higher after fresh embryo transfer following IVM compared to OS (57/122 (46.7%) vs 53/305 (17.4%), P < 0.001), while the results were comparable after FET (63/207 (30.4%) vs 60/166 (36.1%), respectively, P = 0.24). In the multivariate logistic regression analysis evaluating fresh embryo transfer cycles, IVM was the only independent factor (adjusted odds ratio (aOR) 4.24, 95% CI 2.44-7.37, P < 0.001)) significantly associated with increased odds of EPL. On the other hand, when the same model was applied to FET cycles, the type of treatment (IVM vs OS) was not significantly associated with EPL (aOR 0.73, 95% CI 0.43-1.25, P = 0.25). The current data are limited by the retrospective nature of the study and the potential of bias due to unmeasured confounders. The increased risk of EPL after fresh embryo transfer following IVM may point towards inadequate endometrial development in IVM cycles. Adopting a freeze-all strategy after IVM seems more appropriate. Future studies are needed to ascertain the underlying cause of this observation. The Clinical IVM research has been supported by research grants from Cook Medical and Besins Healthcare. All authors declared no conflict of interest. N/A.

Outcome of in-vitro oocyte maturation in patients with PCOS: does phenotype have an impact?

Outcome of in-vitro oocyte maturation in patients with PCOS: does phenotype have an impact?

The Efficacy of Traditional Chinese Medicine Shoutai Pill Combined with Western Medicine in the First Trimester of Pregnancy in Women with Unexplained Recurrent Spontaneous Abortion: A Systematic Review and Meta-Analysis.

Background Shoutai Pill (STP), a famous classic herbal formula documented in traditional Chinese medicine (TCM), is widely available in China for treating unexplained recurrent spontaneous abortion (URSA). This systematic review and meta-analysis aims at evaluating the efficacy and safety of STP in the first trimester of pregnancy in women with a history of unexplained recurrent spontaneous abortion. Methods The following eight databases were searched from their establishment to Dec 31, 2019, for randomized controlled trials (RCTs): PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), the Chinese BioMedical database (CBM), Chinese Scientific Journal Database (VIP), and the Wanfang database. The quality of evidence was estimated by the Grading of Recommendations Assessment, Development, and Evaluation (GRADE). Results A total of 12 studies (916 patients) with URSA were contained in this meta-analysis. The forest plot showed that patients treated with Shoutai Pill and western medicine had a significantly lower incidence of early pregnancy loss (RR: 0.42; 95% CI: 0.34–0.52; P < 0.01, I2 = 0%). Subgroup analysis revealed that different types of TCM syndrome differentiation had the similar results. Also, in the combined group, patients had a lower TCM syndromes and symptoms and serum D-dimer level, while higher incidence of live birth. Conclusions Our findings suggest that cotreatment with STP and western medicine might be superior to western medicine alone in the first trimester of pregnancy to prevent miscarriage in women with unexplained recurrent spontaneous abortion, and there was no adverse event in the experimental group reported. However, the methodological quality of included RCTs was unsatisfactory; it is necessary to verify its effectiveness with further more standardized researches of rigorous design.

Evaluating the effect of ovine placental extract on early embryonic development and incidence of early pregnancy loss after embryo transfer in recipient mares

Advanced Glycation End-products (AGEs) are elevated in the uterine environment of obese infertile women. Can their detrimental effects on endometrial epithelial cells be mitigated with therapeutics, and recapitulated in a more physiologically relevant primary model (organoids)?Human endometrial epithelial cells (ECC-1) were exposed to AGEs at concentrations physiologically representative of the lean and obese uterine fluid, and three potential therapeutics: 25 nM RAGE antagonist FPS-ZM1, 100 μM metformin, and a combination of antioxidants (10 μM N-acetyl-L-cysteine, 10 μM N-acetyl-L-carnitine, and 5 μM α-lipoic acid). Real time cell analysis (xCelligence) determined the rate of adhesion and proliferation. Proliferation of organoid derived cells, and secretion of cytokines from organoids was characterized in the presence of AGEs (n=5). Uterine fluid of women undergoing assisted reproduction was profiled for AGEs-associated inflammatory markers (n ≥ 67 for each correlation).ECC-1 proliferation was reduced by obese versus lean AGEs and vehicle control (P = 0.04), and restored to lean conditions by antioxidants. AGEs influenced organoid derived primary endometrial epithelial cell proliferation in a donor dependent manner. AGEs increased organoid secretion of proinflammatory cytokines, including CXCL16 (P < 0.01). Clinically, CXCL16 correlated positively to maternal BMI (R = 0.26, P = 0.02) and intrauterine glucose (R = 0.74, P < 0.0001).Physiologically relevant concentrations of AGEs alter endometrial epithelial cell function. Antioxidants restore the rate of proliferation of AGEs-treated endometrial epithelial cells (ECC-1). Primary endometrial epithelial cells, cultured as organoids, demonstrate altered proliferation and cytokine secretion in the presence of AGEs equimolar with the obese uterine fluid.

The Role of Metformin for Prevention of First Trimesteric Miscarriage in Women With Polycystic Ovary Syndrome: A Randomized Controlled Trial

Background: This study aimed to evaluate the effect of metformin on early pregnancy loss (EPL) in pregnant women with olycystic ovary syndrome (PCOS).Patients and Methods: A randomized clinical trial (Clinicaltrials.gov NCT 02498522) was conducted in Ain-Shams Maternity Hospital in the period between August 2018 and July 2019. One hundred and sixty six women previously diagnosed with PCOS and got pregnant with induction of ovulation in concomitance with Metformin were randomized either Metformin 500 mg orally every 8 hours until the end of the 1st trimester or not receiving Metformin. The primary outcome is to observe any significant decline in the incidence of EPL in PCOS patients after administration of Metformin through out 1st trimester.Results: There was a significant reduction in miscarriage rate after administration of Metformin during 1st trimester. Introduction of Metformin helped pregnancy loss rate to drop to 10.8% while the rate in the group discontinued Metformin was 42.2% (p < 0.05). The side effects rate as nausea, vomiting, gastric irritation and flatulence was significantly higher in the Metformin group.Conclusion: The use of Metformin in patients with PCOS during 1st trimester reduces the incidence of early pregnancy loss (EPL).

EP16.06: Septate and dysmorphic uterus: association with infertility and early pregnancy loss

To diagnose septate or dysmorphic uterus using three dimensional ultrasound (3DUS), following the European Society of Human Reproduction and Embryology European Society for Gynecological Endoscopy (ESHRE ESGE) consensus on the classification of congenital anomalies of the female genital tract and study their association with pregnancy complications and infertility. Thirty women with uterine anomalies suspected after examination by conventional two dimensional ultrasound were evaluated with 3D US. These data were analysed retrospectively to confirm the presence and type of uterine malformation in accordance with the ESHRE ESGE consensus. Detailed reproductive history was taken and follow up for reproductive outcome. Out of 30 women 25 patients had septate uterus (83.33%) and 5 patients had dysmorphic uterus (16.66%). Early pregnancy loss was seen in 20 patients (66.66%), 10 patients had endometriosis with infertility (33.33%). Our study shows increased incidence of early pregnancy loss and infertility in patients with septate or dysmorphic uterus. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Advanced mare age impairs the ability of in vitro‐matured oocytes to correctly align chromosomes on the metaphase plate

SummaryBackgroundAdvanced mare age is associated with declining fertility and an increased risk of early pregnancy loss. Compromised oocyte quality is probably the primary reason for reduced fertility, but the defects predisposing to embryonic death are unknown. In women, advanced age predisposes to chromosome segregation errors during meiosis, which lead to embryonic aneuploidy and a heightened risk of miscarriage.ObjectivesTo evaluate the effect of advanced mare age on chromosome alignment and meiotic spindle morphology in in vitro‐matured (IVM) oocytes.Study designMorphometric and morphological analysis.MethodsTo investigate differences in spindle organisation and chromosome alignment between young and old mares, oocytes collected from slaughtered mares were divided into two groups depending on mare age (young, ≤14 years and old, ≥16 years), IVM and stained to visualise chromatin and alpha‐tubulin. Spindle morphology, morphometry and chromosome (mis)alignment were evaluated by confocal microscopy and 3D image analysis.ResultsOocytes from old mares showed a higher incidence of chromosome misalignment (47.4% vs. 4.5%; P<0.001) and a thicker metaphase plate (mean ± s.d.: 5.8 ± 1.0 μm vs. 4.9 ± 0.9 μm; P = 0.04) than oocytes from young mares. Although no differences in spindle morphometry were detected between old and young mares, an increased major spindle axis length was associated with chromosome misalignment (mean ± s.d.: 25.3 ± 6.1 μm vs. 20.8 ± 3.3 μm; P = 0.01) irrespective of age.Main limitationsThe oocytes were IVM and may not exactly reflect chromosome misalignment in vivo.ConclusionsAdvanced mare age predisposes to chromosome misalignment on the metaphase II spindle of IVM oocytes. The compromised ability to correctly align chromosomes presumably predisposes to aneuploidy in resulting embryos and thereby contributes to the age‐related decline in fertility and increased incidence of early pregnancy loss. The Summary is available in Portuguese – see Supporting Information

Biomarkers of preconception stress and the incidence of pregnancy loss.

Are biomarkers of preconception stress associated with pregnancy loss? Preconception stress, as measured by basal salivary cortisol and alpha-amylase concentrations, is not associated with pregnancy loss. Many studies, most of which have been retrospective, have identified an association between stressful life events and perceived stress and miscarriage. A prospective pregnancy study with preconception enrollment was conducted between 2005 and 2009. Among the 344 women who became pregnant during the Longitudinal Investigation of Fertility and the Environment (LIFE) study, 337 (98%) had salivary biomarker data for analysis. Couples planning pregnancy were followed for up to 12 months as they tried to become pregnant and through pregnancy if it occurred. Participating women collected a basal saliva sample on the morning following enrollment and a second on the morning following their next menses to measure cortisol and alpha-amylase, biomarkers of stress. Women used home pregnancy tests on the day of expected menses. A pregnancy loss was defined as a negative pregnancy test following a positive pregnancy test, the onset of menses, or for pregnancies that survived to clinical recognition, recognition of the loss by a healthcare provider. Among the 337 couples, the median age of female and male partners was 29 and 31 years, respectively. Most of the women were non-Hispanic white (83%) and highly educated. There were 97 pregnancy losses reported among the 337 pregnancies. The median gestational age at loss was 6 weeks 5 days with only two losses occurring in the second trimester. Using Cox proportional hazards models, we found no clear pattern of association between two preconceptional biomarkers of stress (salivary cortisol and alpha-amylase concentrations) modeled both continuously or in tertiles and incident pregnancy loss after adjustment for confounders. Our prior work suggests that women enrolled in the LIFE Study had lower stress levels than women in the general population. Owing to concerns regarding participant burden, we were unable to collect serial saliva measurements, which would have allowed us to examine the association between stress in early pregnancy and pregnancy loss. Further, with regard to the measurement of perceived stress, the Cohen's Perceived Stress Scale was only administered at baseline. While every attempt was made to ensure diversity in the cohort, non-Hispanic white women were over-represented, therefore it is possible that the results might not be generalizable to all women. In one of the largest studies in the USA to prospectively capture data on the incidence of early pregnancy loss, we found no clear association between two biomarkers of preconception stress (measured in saliva) and pregnancy loss. This study was supported by the Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development (contracts #N01-HD-3-3355, N01-HD-3-3356, N01-HD-3358). There are no conflicts of interest to declare. Not applicable.

Signs and symptoms associated with early pregnancy loss: findings from a population-based preconception cohort.

What is the relationship between signs and symptoms of early pregnancy and pregnancy loss <20 weeks' gestation? Vaginal bleeding is associated with increased incidence of early pregnancy loss, with more severe bleeding and bleeding accompanied by lower abdominal cramping associated with greater incidence of loss; conversely, vomiting is associated with decreased incidence of early pregnancy loss, even in the setting of vaginal bleeding, while nausea alone is not. Two previous cohort studies with preconception enrollment suggested that bleeding is associated with loss while nausea is inversely associated with loss though these studies were limited by small study size and reporting after loss ascertainment. No prior preconception cohort study has examined multiple signs and symptoms in relation to pregnancy loss. Population-based preconception cohort of 501 couples discontinuing contraception to try for pregnancy in 16 counties in Michigan and Texas, USA. Participants were followed daily until positive home pregnancy test or 12 months of trying without an hCG pregnancy; women who became pregnant were followed daily from 2 to 7 weeks post-conception. Three hundred and forty-seven women had a positive home pregnancy test denoting hCG pregnancy. Three hundred and forty-one women remained after excluding ineligible pregnancies. Women recorded daily from 2 to 7 weeks post-conception their signs and symptoms, including vaginal bleeding (none, spotting, light, moderate and heavy), lower abdominal cramping, nausea and vomiting. Pregnancy losses were ascertained by a subsequent negative home pregnancy test, clinical confirmation or onset of menses, depending on gestational age at loss; time-to-loss was measured in days post-conception. Cumulative incidence functions and 95% confidence intervals (CIs) were constructed for each sign or symptom, and hazard ratios (HRs) and 95% CIs for presence compared with absence of signs or symptoms were estimated using Cox proportional hazard models. Women experienced lower abdominal cramping (85%), nausea (48%), vomiting (46%) and light/moderate/heavy vaginal bleeding (24%) during early pregnancy. Ninety-five (28%) women experienced a loss. Cumulative incidence of pregnancy loss varied by symptomatology: 19% for vomiting, 27% for lower abdominal cramping, 35% for nausea only, 52% for vaginal bleeding, 81% for vaginal bleeding with lower abdominal cramping. Incidence of pregnancy loss was increased among women with vaginal bleeding (HR: 3.62, 95% CI: 2.29-5.74) and among women with vaginal bleeding and lower abdominal cramping (HR: 5.03, 95% CI: 2.07-12.20). Incidence of pregnancy loss was decreased for women with vomiting (HR: 0.51, 95% CI: 0.30-0.86). In the setting of vaginal bleeding with lower abdominal cramping, vomiting reduced the incidence of pregnancy loss (HR: 0.24, 95% CI: 0.11-0.56). There were few losses beyond 14 weeks gestation; thus, the precision of our findings related to losses occurring after the first trimester is limited. By using sensitive home pregnancy tests, we are able to document and characterize the cumulative incidence of the earliest pregnancy losses, which constitute the majority of losses. The use of daily, prospective capture of signs and symptoms relative to ascertainment of pregnancy loss minimizes potential biases associated with reporting after rather than before a loss, which could potentially distort the relationship between signs and symptoms and pregnancy loss. The findings of our study suggest that it may be useful to develop prognostic models for pregnancy loss based on signs and symptoms. This study was supported by the Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health (contract numbers N01-HD-3-3355; N01-HD-3-3356; N01-HD-3-3358). The authors have no conflict of interest to declare.

Deutsche Erstzucht des Halsbandmakis (Eulemur collaris)

In this paper, we described the incidence of early pregnancy loss (EPL) both after natural mating and embryo transfer, evaluated risk factors, and summarized the outcome of twin pregnancies throughout gestation in dromedaries under reproductive care. Data were collected over seven breeding seasons at the world's largest camel dairy farm (study 1). In addition, we determined the timing of EPL and monitored serum progesterone (P4) concentration between Days 13–70 of gestation during one breeding season (study 2). In the first study, out of 2970 pregnancies, 507 cases (17.1%) of EPL were diagnosed with transrectal ultrasonography. The rate of EPL after natural mating and embryo transfer was 16.1% (n = 422 out of 2616) and 24.0% (n = 85 out of 354), respectively. Twin pregnancies were detected in 215 cases (7.2% of all gestations), and 57 of those (26.5%) underwent complete EPL. Almost half of the early losses (n = 243; 47.9%) occurred before 30 d of gestation. Another 43.2% (n = 219) of EPL was diagnosed during the next month, and 8.9% (n = 45) occurred after 60 d of gestation. Multivariable mixed effects logistic regression models revealed that the breeding season (year) and twin pregnancy were the most important exposure variables affecting the rate of EPL (P < 0.001). The effect of some male camels was also demonstrated while other factors, such as type of breeding, age category, month of mating, breed/ecotype and reproductive history did not prove to have a significant influence. In the second study, the overall rate of EPL was 24.5% (n = 34 of 139). There was no difference in the incidence of EPL between ET recipient (24.2%, n = 23 of 95) and mated (25%, n = 11 of 44) camels. Weekly rate of EPL ranged from 0.9% to 4.8% with a decreasing tendency, and approx. 41% of the animals (n = 14 of 34) had some ultrasonographic signs of impending EPL 1 week before the final diagnosis. Mean serum P4 concentration in camels with subsequent EPL was 5.3 ± 0.1 ng/ml compared to 5.6 ± 0.04 ng/ml in normal pregnant dromedaries. Day of gestation and future EPL influenced serum P4 levels (P < 0.001) with an interaction between the two fixed factors (P < 0.05). At the time of the final diagnosis of EPL, mean serum P4 concentration was 2.8 ± 0.44 ng/ml. Although twinning had an unfavorable prognosis with a total pregnancy loss of 36.7%, it was not entirely detrimental for the final outcome of gestation as two-thirds of twin pregnancies (n = 136 out of 212) resulted in the birth of a live calf.