hCG Levels Research Articles

Assessment of kisspeptin and trappin level in Iraqi women with early pregnancy loss: a cross sectional study

Background: Early pregnancy loss (EPL) is a prevalent medical condition that can have significant and long-lasting impacts on an individual's life. At present, the determination of the feasibility of initial gestation is ascertained through the measurement of serum human chorionic gonadotropin (hCG) concentrations. Notwithstanding, an elevated hCG level in women during their initial trimester who exhibit bleeding does not constantly imply a sustainable pregnancy. Kisspeptin, a well-established regulator of maturation that is widely present in the placenta, represents a valuable biomarker for the assessment of miscarriages and placental dysfunction. The objective of this investigation was to ascertain key biomarkers (namely kisspeptin, trappin, and hCG) in females who have experienced a previous miscarriage. Methods: The study included 90 participants, including 60 pregnant women having a history of abortion in the first trimester and 30 healthy individuals who attended Al Kadhimiya Teaching Hospital and Abu Ghraib Hospital Baghdad Governorate, and ranged in age from 18 to 38 years. Sera from all participants were tested to evaluate various research parameters. Results: Trappin levels were less crucial to hCG than kisspeptin levels. The risk of miscarriage decreases as kisspeptin levels increase. Conclusions: The development of a preliminary screening test aimed at identifying patients who are at risk of miscarriage could potentially offer benefits in terms of offering supplementary emotional assistance and continuation of medical care. Kisspeptin has been identified as a prospective indicator for placental performance. Kisspeptin exhibits potential as a biomarker for assessing the viability of pregnancy and holds promise for clinical utility in the development of a precise diagnostic tool for early pregnancy outcome prediction.

Efficacy of estrogen-progestogen therapy for women with vascular retained products of conception following miscarriage or abortion.

To assess the clinical characteristics and endocrinological background of women with vascular retained products of conception (RPOC) after miscarriage or abortion and evaluate the effect of estrogen-progestogen therapy (EPT) as an initial treatment on this population based on their endocrinological background. Women with vascular RPOC after miscarriage or abortion at less than 20 weeks of pregnancy who were given EPT (conjugated estrogen and norethisterone) were retrospectively reviewed. Their clinical characteristics, hormonal parameters, ultrasonographic findings, and outcomes were evaluated. Of 35 women with vascular RPOC, 30 (86%) presented with vaginal bleeding at a visit, and 6 (17%) required inpatient management due to heavy bleeding. Among women who presented with vaginal bleeding, serum progesterone levels were significantly lower (0.25 vs. 6.5ng/mL, p=0.004) than those in women who did not present with vaginal bleeding. There were no differences in serum hCG levels (10.5 vs. 3.1 mIU/mL) or serum estradiol levels (65.4 vs. 162.3pg/mL). After withdrawal bleeding following the first course of EPT, vaginal bleeding was stopped in 27 of the 30 women (90%), and 23 (66%) of all women had a thin and linear endometrium. All women could be treated by up to two courses of EPT and did not require additional interventions. The median duration to hCG normalization after the initial EPT was 24.5 (9-88) days. Women with vascular RPOC who have no bleeding had significantly higher levels of serum progesterone, indicating that administration of progestogen may have an effect on hemostasis. Endometrial bleeding can be prevented or stopped, and retained tissues can be conservatively expelled by oral administration of EPT, including norethisterone, in women with vascular RPOC.

Association of serum beta hCG levels in women with palpable malignant breast lesions

This study aims to determine whether serum Beta hCG can be used as a tumour marker in Breast malignancies. The objective of this study is to evaluate the serum Beta hCG in various stages of breast carcinoma and to correlate its level with disease severity and prognosis. Cross sectional analytical study of assessing serum Beta hCG in 200 patients with palpable breast malignancies at hospitals in urban Mangalore, India. In our study there was No increase in serum Beta hCG, in women with breast malignancies, but there was a pattern amongst the negative results. A Beta hCG of < 5mIU/mL is taken as negative, but in our study of 200 individuals, a mean value of 2mIU/mL was used as differentiation between low and high risk individuals. With our study we tried to correlate the value of Beta hCG with malignant breast lesions, and even though women with such lesions did not have a value of > 5mIU/mL, we found substantial evidence that women who had a value of > 2mIU/mL had a more advanced disease, be it in terms of staging, and comparing it with markers like ki67. A direct correlation between Beta hCG and severity of the disease in terms of staging was proved, hereby directly affecting the outcome of patients. Higher the level of Beta hCG, graver the prognosis. Even though Beta hCG cannot be used as tumour marker, it can be used to prognosticate the severity in women with palpable breast malignancies.

Exposure to a phthalate mixture disrupts ovulatory progesterone receptor signaling in human granulosa cells in vitro†.

Exposure to phthalates disrupts ovarian function. However, limited studies have investigated the effects of phthalate mixtures on ovulation, especially in women. Human granulosa cells were used to test the hypothesis that exposure to a phthalate mixture (PHTmix) disrupts progesterone (P4)/progesterone receptor (PGR) signaling, which is a crucial pathway for ovulation. In addition, progestin and cyclic adenosine 3', 5'-monophosphate (cAMP) supplementation were tested as methods to circumvent phthalate toxicity. Granulosa cells from women undergoing in vitro fertilization were acclimated in culture to regain responsiveness to human chorionic gonadotropin (hCG; clinical luteinizing hormone analogue). Granulosa cells were treated with or without hCG, and with or without PHTmix (1-500μg/ml; dimethylsulfoxide = vehicle control) for 0.5-36h. In the supplementation experiments, cells were treated with or without R5020 (stable progestin), and with or without 8-Br-cAMP (stable cAMP analogue). Exposure to hCG + PHTmix decreased P4 levels and mRNA levels of steroidogenic factors when compared to hCG. This was accompanied by decreased mRNA levels of PGR and downstream P4/PGR ovulatory mediators (ADAM metallopeptidase with thrombospondin type 1 motif 1 (ADAMTS1), C-X-C motif chemokine receptor 4 (CXCR4), pentraxin 3 (PTX3), and regulator of G protein signaling 2 (RGS2)) in the hCG + PHTmix groups compared to hCG. Exposure to hCG + PHTmix 500μg/ml decreased cAMP levels and protein kinase A activity compared to hCG. Supplementation with progestin in the hCG + PHTmix 500μg/ml group did not rescue toxicity, while supplementation with cAMP restored PGR levels and downstream P4/PGR mediator levels to hCG levels. These findings suggest that phthalate mixture exposure inhibits P4/PGR signaling in human granulosa cells via decreased steroidogenesis, cAMP levels, and protein kinase A activity. Restored P4/PGR signaling with cAMP supplementation provides a potential cellular target for intervention of phthalate-induced ovulatory dysfunction in women.

Twelve Hours Post-Injection Serum Human Chorionic Gonadotropin and Body Mass Index Predicts In Vitro Fertilization Oocyte Maturation Rate: A Cross-Sectional Study.

In vitro fertilization (IVF) remains a main treatment for infertility cases. Post-injection human chorionic gonadotropin (hCG) level is an essential factor in determining oocyte maturation rate in IVF. This study aimed to determine the relationship between 12 hours post-injection serum hCG level and oocyte maturation rate among IVF participants. A cross-sectional study on IVF participants was done at a tertiary hospital in Indonesia from January 2020 to December 2021. Subjects were injected with 250 μg of recombinant-hCG (r-hCG) subcutaneously. Twelve hours post-injection serum hCG levels and oocyte maturation rate data were retrieved and analyzed accordingly. Twenty-eight subjects were recruited into the study. Higher 12 hours post-injection serum hCG was related to oocyte maturation rate (P=0.046). The cut-off point of 12 hours post-injection serum hCG to predict better oocyte maturation rate was 90.15 mIU/mL (sensitivity 68.2%, specificity 83.3%). Oocyte maturation rate may be predicted using body mass index (BMI) and 12 hours post-injection serum hCG. Higher 12 hours post-injection serum hCG was associated with a higher oocyte maturation rate in IVF subjects.

Beta hCG, urea and creatinine levels in vaginal fluids: a reliable marker for prelabour rupture of membranes

Beta hCG, urea and creatinine levels in vaginal fluids: a reliable marker for prelabour rupture of membranes

Prediction of Treatment Option by Human Chorionic Gonadotropin (hCG) Levels in Ectopic Pregnancy using Machine Learning

Aim: The objective of this study is to construct a model using a random forest to predict the treatment option of ectopic pregnancy based on hCG levels, as well as to confirm the model’s accuracy. Methods: We selected 17 variables related to ectopic pregnancy and extracted data from our records for cases of possible ectopic pregnancy. We then divided the cases into two groups: 1) laparoscopic surgery and 2) MTX or conservative treatment. We created a model for predicting the prognosis of ectopic pregnancy. Afterward, we confirmed the model’s accuracy using the test data. Additionally, we compared the model’s accuracy with that of two specialized obstetriciangynecologists (OB/GYNs) specialists who judged the same data. This study was approved by our ethics committee. Results: One hundred and twenty-eight patients were eligible for this research, of whom 52.3% (67) underwent laparoscopic surgery and 7.0% (9) had emergent laparoscopic surgery. MTX and conservative treatment, including normal pregnancies or miscarriages, were 25.0% (32) and 25.0% (32), respectively. The model’s accuracy using a random forest was 87.3%, and the area under curve (AUC) was 0.784. The two OB/GYNs judged the same data with respective accuracies of 77.3% and 79.7%. Conclusion: In conclusion, this model using a random forest was superior to the judgment of specialists. Moreover, this research is new in the fact that it has presented a numerical model involving multiple risks that, until now, have been judged empirically by humans. In the future, it may help develop and elucidate a more extensive prediction system for ectopic pregnancy.

Comparison of HCG Levels in Reagents Temperature 25°C and 8°C Using ELISA Method

HCG hormone is a hormone produced by the placenta in early pregnancy with the aim of maintaining pregnancy. The Elisa method is a method for examining HCG levels which has the principle of specific antigen and antibody reactions. The work stage in the laboratory that has the biggest error is in the pre-analytic stage reaching 60%, one of which is the reagent temperature. The ELISA method recommends the reagent temperature to be stored at 2-8°C and when used at 25°C. Examination reagents Elisa method uses reagents that use enzymes in the examination. Enzymes are very sensitive to temperature. This study aims to determine the ratio of HCG levels at reagent temperatures of 25°C &amp; 8° using the ELISA method. This type of research is analytic supported by experiments and literature studies. Serum samples of 6 pregnant women were taken from the Kedungmundu Public Health Center using a limited population count formula. The examination was carried out by comparing the reagents at a temperature of 25°C and 8°C using 6 serums of pregnant women and in duplicate. Data analysis only used primary data and statistical tests were carried out using normality, homogeneity and T-Test tests. The average result at a temperature of 25°C was 380.851 and the average temperature of 8°C was 321.99 and the T-test statistic test showed p value = 0.469 and 0.470. Based on the results of the study, it can be concluded that there is an insignificant difference in the concentration of HCG by comparing the reagents at 25°C and 8°C.

GDF-11 downregulates placental human chorionic gonadotropin expression by activating SMAD2/3 signaling

BackgroundThe production of human chorionic gonadotropin (hCG) by the placental trophoblast cells is essential for maintaining a normal pregnancy. Aberrant hCG levels are associated with reproductive disorders. The protein of hCG is a dimer consisting of an α subunit and a β subunit. The β subunit is encoded by the CGB gene and is unique to hCG. Growth differentiation factor-11 (GDF-11), a member of the transforming growth factor-β (TGF-β) superfamily, is expressed in the human placenta and can stimulate trophoblast cell invasion. However, whether the expression of CGB and the production of hCG are regulated by GDF-11 remains undetermined.MethodsTwo human choriocarcinoma cell lines, BeWo and JEG-3, and primary cultures of human cytotrophoblast (CTB) cells were used as experimental models. The effects of GDF-11 on CGB expression and hCG production, as well as the underlying mechanisms, were explored by a series of in vitro experiments.ResultsOur results show that treatment of GDF-11 downregulates the expression of CGB and the production of hCG in both BeWo and JEG-3 cells as well as in primary CTB cells. Using a pharmacological inhibitor and siRNA-mediated approach, we reveal that both ALK4 and ALK5 are required for the GDF-11-induced downregulation of CGB expression. In addition, treatment of GDF-11 activates SMAD2/3 but not SMAD1/5/8 signaling pathways. Moreover, both SMAD2 and SMAD3 are involved in the GDF-11-downregulated CGB expression. ELISA results show that the GDF-11-suppressed hCG production requires the ALK4/5-mediated activation of SMAD2/3 signaling pathways.ConclusionsThis study not only discovers the biological function of GDF-11 in the human placenta but also provides important insights into the regulation of the expression of hCG.EYYrMgGK1oNKot-fDMBUfiVideo

SARS-CoV-2 in early pregnancy-does it affect the aneuploidy screening markers and cause pregnancy loss?

It is known that vertical transmission of various infections poses a potential risk to the fetus, especially in early pregnancy. Potential effects of SARS-CoV-2 infection on early pregnancy and placental formation and functions still remain unknown. To determine the alterations of prenatal aneuploidy screening markers in a group of pregnant women who were SARS-CoV-2 positive during the first trimester. The secondary goal was to assess pregnancy loss rates. The study group consisted of pregnant women who were diagnosed with mild forms of SARS-CoV-2 infection before the screening test at any time in early pregnancy. The control group included pregnant women who were not diagnosed with SARS-CoV-2 infection during their pregnancy. SARS-CoV-2 infection was detected by RT-PCR in the nasopharyngeal swab samples. Multivariate linear regression analysis was performed due to evaluate effect of SARS-CoV-2 infection on NT and serum aneuploidy screening parameters taking maternal age and gestational age which the COVID-19 RT-PCR test result was positive into account. We did not find any significant difference between the COVID-19-positive and COVID-negative groups in gestational age at screening, sonographic measurements of CRL, NT, and serum levels of PAPP-A, free hCG, and triple test serum markers even after accounting for maternal age and gestational age which the COVID-19 RT-PCR test result was positive. There was no statistically significant difference in pregnancy loss. We did not find any evidence for unfavorable prenatal biochemical, ultrasound markers of fetal aneuploidy screening tests, and pregnancy loss rates in our study group.

Analysis of mid-trimester maternal serum β-hCG and AFP as markers of preterm and term adverse pregnancy outcomes form a tertiary care hospital, Morang, Nepal

Background: Mid-trimester maternal serum markers have been used for prenatal aneuploidy screening for a long time. The aim of the study was to assess the mid-term serum levels of β-human chorionic gonadotropin and alpha-fetoproteins for placenta-mediated adverse pregnancy outcomes (PMAPOs) in preterm and term pregnancies.&#x0D; Material and methods: A prospective cohort study involving nulliparous women with singletons without aneuploidy or fatal fetal abnormalities was carried out a tertiary care hospital, Morang, Nepal. AFP and β-hCG levels were estimated between 13 and 17 weeks of gestation in the mother's serum. All values were in multiples of the median (MoM) and compared between women with PMAPOs.&#x0D; Results: The serum levels of AFP and β-hCG were obtained in 176 out of 300 nulliparous women. The MoM of serum β-hCG (1.3 vs 1.1) and AFP (1.4 vs 1.1) were higher in PMAPOs-affected women than in controls.&#x0D; Conclusion:Preterm PMAPOs, but not term PMAPOs, are more likely in the present study when maternal serum AFP or β -hCG levels are more significant than 2.0 MoM. If fetal growth is within the normal range at 37 weeks of gestation, it is advisable that women with increased serum β-hCG or AFP receive regular prenatal care.

Impact of trophoectoderm biopsy for preimplantation genetic testing on serum β-hCG levels, time of delivery and birthweight following frozen embryo transfer cycles

Aim: This study investigated whether trophoectoderm (TE) biopsy adversely impacts serum β-human chorionic gonadotropin (hCG) level on the 15th day of embryo transfer (ET), delivery week and birthweight, between biopsied and unbiopsied embryo groups, in a cohort of women who delivered a singleton baby, following frozen–thawed ET. Methods: All women having had a live birth after blastocyst ETs following frozen ET cycles with preimplantation genetic testing (PGT) were included. A control group was selected among women who had a live birth following single frozen blastocyst transfer without PGT-A at the same period in our clinic Results: One hundred fifteen and 173 cycles with- and without-PGT, respectively, were included. Serum β-hCG level on the 15th day after ET was comparable between the groups (p = .336). Average birthweight of the babies born following biopsied embryos were significantly lower (3200 vs. 3380; p = .027). Women who received trophectoderm biopsied embryos had a significantly higher probability of having a baby weighing ≤1500 g and 1500–2500 g (p = .022) or ≤2500 g (p = .008). Proportion of preterm delivery was significantly higher in the biopsy group (p = .023). However, after adjusting for potential covariates, trophectoderm biopsy did not seem to increase the risk of preterm birth (OR 1.525; 95% CI, 0,644–3.611; p = .338) Conclusions: TE biopsy does not seem to impact serum β-hCG level on the 15th day after ET. Average birthweight is lower when a biopsied embryo was transferred. After adjusting for potential covariates, trophectoderm biopsy does not seem to increase the risk of preterm birth

P-363 Ovarian endometriosis deteriorates embryo development in ART cycles: An embryonic time-lapse study

Abstract Study question Does ovarian endometriosis deteriorate embryo quality in terms of morphokinetic parameters determined by time-lapse images? Summary answer Embryos from ovarian endometriosis patients accelerate their early developmental events but decelerate after t5. Completion of late developmental events were significantly delayed in endometriosis group. What is known already Endometriosis may affect the oocyte quality and consequently affect the embryo quality in terms of embryo morphology and implantation ability. Morphokinetic evaluation of embryos provided more developmental details and might be utilised as a tool to distinguish the embryo developmental ability. Only scarce studies with limited case numbers have evaluated the association between endometriosis and developments of embryo in terms morphokinetic observation. Hardly any consensus has ever been drawn. Study design, size, duration The morphokinetic parameters of embryos were compared between patients with or without ovarian endometriosis. A total of 52 ovarian endometriosis cases and 208 controls without endometriosis, receiving their assisted reproductive techniques from April 2020 through June 2022, were included for analysis. Propensity Score Matching with age, BMI and AMH was performed in a ratio of 1:4 in order to eliminate the confounding factors. Participants/materials, setting, methods We analyzed a total of 737 top quality embryos obtained from 348 oocyte retrieval cycles performed at Taipei Fertility Center. 127 embryos were obtained from women affected by ovarian endometriosis and 617 were obtained from unaffected controls. All embryos were cultured in a time-lapse incubator chamber and completed blastulation. Main results and the role of chance Morphokinetic data showed that embryo development is distorted in embryos obtained from women with endometriosis. The timing of early developmental events, including tPNf (22.7 +/- 2.9 hr vs 23.5 +/- 4.1 hr, p = 0.03) and t2 (25.2 +/- 3.1 hr vs 26.1 +/- 4.4 hr, p = 0.03) were significantly shortened in the endometriosis group. However, the timing of late developmental events, including tM (89.7 +/- 9.8 hr vs 86.6 +/- 11.4 hr, p = 0.006) and tB (111.0 +/- 11.0 hr vs 108.3 +/- 12.5 hr, p = 0.03) were significantly prolonged in the endometriosis group. The timing of t5 by morphokinetic observation was found to be equal both in ovarian endometriosis group and control group (50.4 +/- 8.7 hr vs 50.4 +/- 8.4 hr, p = 0.99). The average of KID score of embryos from ovarian endometriosis group was lower than that from normal control, but it did not reached statistical significance (4.07 +/- 2.11 vs 4.49 +/- 2.70, p = 0.27). The initial serum hCG levels in pregnant women from both endometriosis and non-endometriosis participants showed that hCG levels were significantly higher in non-endometriosis cases (628.8 +/- 967.7 mIU/ml vs 1581.5 +/- 1885.1 mIU/ml, p = 0.01). Limitations, reasons for caution This is a retrospective observational study with propensity score matching method to eliminate confounding factors such as age, BMI and AMH. Wider implications of the findings The effects of prolonged morphokinetic parameters on the embryo quality need further study to clarify; however, they may serve as markers in selecting embryos with implantation potential. Trial registration number Not applicable

O-024 Exploring the optimal transfer day of day 6 (D6) vitrified blastocysts in Hormone Replacement Therapy Frozen Embryo Transfer (HRT-FET) cycles - a cohort study

Abstract Study question Should blastocysts cryopreserved D6 be transferred on the 5th or 6th day of progesterone administration in HRT-FET cycles? Summary answer No significant differences in outcome parameters were found when D6 vitrified blastocysts were transferred on either day 5 or day 6 of progesterone in HRT-FET. What is known already Several studies have shown that D6 blastocyst transfer should be avoided in fresh transfer IVF cycles; consequently, these more slowly developing embryos are transferred in a HRT-FET cycle on the 6th day of progesterone administration in line with day 5 (D5) blastocysts. For successful implantation, a receptive endometrium and optimal synchronization between the endometrium and embryo are important. However, no definitive evidence exists regarding the optimal timing of transferring D6 blastocysts. Interestingly, a few studies have reported better reproductive outcomes if D6 blastocysts are transferred on the 5th day of progesterone administration instead of the 6th day of progesterone administration. Study design, size, duration This cohort study included a total of 718 vitrified D6 single blastocyst HRT-FET cycles performed between 2013 and 2022. 576 blastocysts were transferred on the 6th day of progesterone administration (2013-2021), and 142 blastocysts transferred on the 5th day of progesterone administration (2021-2022). Main outcomes were biochemical pregnancy rate (PR), implantation rate (IR), clinical pregnancy rate (CPR), and early pregnancy loss rate (EPLR). Participants/materials, setting, methods Patients undergoing HRT-FET in a public Fertility Clinic. Endometrial preparation included oral oestradiol (6mg/24hours), followed by vaginal micronized progesterone. As mentioned above, single blastocyst transfer was performed on either the 5th or 6th day of progesterone administration. Serum β-hCG level was measured 9-11 days after embryo transfer and biochemical pregnancy was defined as an hCG level &amp;gt;10 IU/L. Clinical pregnancy was defined as visualization of a gestational sac and fetal heartbeat in gestational week 7. Main results and the role of chance In this cohort of a total of 718 HRT-FET cycles transferred on either the 5th or 6th day of progesterone administration the mean age of the patients at embryo transfer was 34.5±4.5 years vs. 33.0±4.9 years, respectively, p = 0.30. There was no significant difference between numbers of top-quality blastocyst transfers within the two groups 63% (89/142) vs. 64% (369/576), p = 0.76. The overall, biochemical pregnancy rate (PR), implantation rate (IR), clinical pregnancy rate (CPR) and early pregnancy loss (EPLR) in the group of patients transferred on the 5th day of progesterone administration were 38% (54/142), 29% (41/142), 25% (36/142) and 33% (18/54). In the group of patients transferred on the 6th day of progesterone administration 41% (235/576), 33% (191/576), 30% (170/576) and 28% (65/235). There were no significant differences between PR, IR, CPR and EPLR when transferring D6 vitrified blastocysts on either the 5th or the 6th day of progesterone administration (38% vs. 41%, p = 0.53, 29% vs. 33%, p = 0.33, 25% vs. 30%, p = 0.33 and 33% vs. 28%, p = 0.41). Limitations, reasons for caution Limitations of the study are the retrospective design and the limited number of D6 vitrified blastocysts transferred on the 5th day of progesterone administration. Furthermore, the two groups represented two different time periods. Wider implications of the findings Growing evidence shows that D5 vitrified blastocysts have better reproductive outcomes than slower developing embryos which are vitrified on day 6. Whether this is caused by intrinsic embryonic factors or poor timing of transfer in a subsequent FET cycle still needs to be unraveled. Trial registration number not applicable

P-660 No association between BMI, oocyte yield and maturation following different trigger strategies: a large observational study including 5000 cycles.

Abstract Study question Is patients’ BMI associated with oocyte yield and maturation following different trigger strategies with r-hCG, GnRH agonist (GnRHa) or dual trigger? Summary answer Patients’ BMI is not associated with oocyte yield and oocyte maturation following triggering with rhCG, GnRHa or dual trigger. What is known already Triggering final oocyte maturation is a key step of ovarian stimulation for IVF/ICSI treatment. Although hCG has traditionally been the gold standard for final oocyte maturation, GnRHa is increasingly used as optimal trigger in “freeze-all” era. Recent evidence has demonstrated excellent results with dual-trigger, a combination of hCG and GnRHa. Despite this swift in clinical practice, early studies demonstrated that hCG plasma levels depend on hCG trigger dose and BMI. Furthermore, although dose-finding studies following for GnRHa trigger demonstrated equal efficacy of triptorelin doses ranging from 0.1-0.4mg, these studies pertained only to low BMI patients. Study design, size, duration This is a retrospective observational study including 5190 consecutive cycles ovarian stimulation cycles (OS) performed between January 2019 -September 2022 in a tertiary Fertility Unit within a University Affiliated Hospital. Overall 5190 ovarian stimulation cycles were analyzed: 2691 cycles triggered with subcutaneous administration of 0.2 mg of GnRHa (triptorelin), 1110 with 250 mcg of r-hCG and 1389 with a dual trigger (combination of both 0.2 mg triptorelin+ 250 mcg of r-hCG). Participants/materials, setting, methods Ovarian stimulation was performed with recombinant FSH or HMG at a starting dose 150-300IU depending on patients’ ovarian reserve and BMI. Control of LH surge was accomplished by a flexible GnRH antagonist or PPOS protocol. The primary outcome measures were oocyte maturation rate (MII/oocytes) and FOI (oocytes/AFC); secondary outcomes were oocyte and MII yield. Multivariable regression models were performed to evaluate the interaction between BMI and type of trigger in relation to study’s outcomes. Main results and the role of chance Overall, oocyte and MII yield was significantly different between different types of trigger mainly due the fact that GnRHa was primary used in freeze-all cases, with excellent ovarian reserve. This resulted in a higher number of oocytes (15.95 ± 8.88) and MIIs (12.31 ± 7.27) as compared with cycles triggered with in rhCG (7.83 ± 5.42 and 5.86 ± 4.22 respectively) or dual trigger (8.51 ± 5.70 and 6.44 ± 4.54 respectively) was utilized. Nonetheless, when oocyte maturation rate was comparable between trigger groups (% [95% CI]): GnRHa 77.2 [76.6; 77.8] vs rhCG 74.9 [73.3; 76] vs dual 75.6 [74.5; 76.7]. Multivariable regression analysis, adjusting for confounding factors, demonstrated that BMI was not associated with oocyte maturation rate (OR:1.00 [95% CI: 0.99;1.01]) and FOI (Beta 0.52 [95% CI: -0.49;1.54]). Similarly BMI was not associated number oocytes (Beta 0.02 [95% CI: -0.08; 0.13]) or on the number of MIIs (Beta 0.01 [95% CI: -0.08; 0.10]) retrieved. Interactions between BMI and trigger groups were considered in each model. All analyses were conducted considering patients’ weight, but no association was revealed. Limitations, reasons for caution This is a retrospective study, and we cannot exclude the presence of residual bias despite the large series, the multiple regression models and the indexes like maturation rate and FOI we adopted to account for confounding factors. Wider implications of the findings Although previous small observational studies supported an association between BMI and hCG levels, our large analysis of &amp;gt; 5000 cycles clearly shows that the efficiency of r-hCG, triptorelin or dual trigger is not associated with the patient's BMI and weight, as no significant differences were found in maturation rate and FOI. Trial registration number Not applicable

P-551 Association between the length of in vitro culture and mode of ART and the initial se-hCG rise in ongoing singleton pregnancy

Abstract Study question Does the length in vitro and mode of ART (fresh and frozen cycles) impact the se-hCG rise in cycles with ongoing single implantation pregnancy? Summary answer Length in vitro and mode of ART alter the hCG. Blastocysts in FET induced higher hCG than FET clevage stage embryos and fresh blastocyst transfers. What is known already Human chorionic gonadotropin (hCG) is produced by placental trophoblasts providing the first measurable sign of pregnancy. Factors, such as multiple pregnancies, advanced maternal age and gender of the fetus are known to influence hCG levels. Other factors such as mode of ART and length in vitro have been considered to alter the hCG kinetics but with conflicting results. Some studies have shown that hCG levels are higher after fresh blastocyst transfer compared with fresh cleavage stage embryo transfer (ET). Other studies found frozen-thawed embryo transfer (FET) results in higher hCG compared with fresh ET, other studies concluded the opposite. Study design, size, duration Multicenter historical cohort study based on clinical data. The study included 5271 women undergoing in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), insemination (IUI) or FET with single ET followed by hCG measurement day 15-19 after ovulation induction (IVF/ICSI/IUI/natural cycle FET) or progesterone administration (estrogen/progesterone substituted FET cycles). All included cycles resulting in single implantation and ongoing pregnancy verified by ultrasound in week 7-8. Data was collected between January 2014 and December 2021. Participants/materials, setting, methods Data was prospectively collected from the Danish Medical Data Centre used by three public fertility clinics, Copenhagen University hospitals. The association between length in vitro (no in vitro period (IUI), day2/3, day5, day6) and mode of ART (IVF/ICSI, FET) and the initial hCG rise was tested both with linear regression and multivariable regression. The results were based on mean serum hCG and presented in percentage with the corresponding 95% confidence interval (CI). Main results and the role of chance After accounting for the pre-defined exclusion criteria (i.e. oocyte donation, pre-implantation genetic testing, more than one gestational sac), 5251 cycles were included in the study. 2122 were FET, 2521 were fresh ET and 608 were IUI. The initial hCG rise was overall lower for fresh ET compared to IUI (no in vitro period). For cleavage stage ET, hCG was 18% (95% CI: 13% - 23%, p &amp;lt; 0.001) lower, and for blastocyst transfer 23% (95% CI: 18% - 28%, p &amp;lt; 0.001) lower. In FET, hCG was 26% (95% CI: 13% - 40%, p &amp;lt; 0.001) higher for blastocyst transfer compared to cleavage stage ET. When comparing blastocyst transfer in FET vs. fresh ET cycles, hCG was 33% (95% CI: 27% - 45%, p &amp;lt; 0.001) higher in FET cycles. Stratifying FET cycles in natural and substituted cycle did not alter the result. All results remained significant after adjusting for referral diagnosis, women age and treatment center. Limitations, reasons for caution It cannot be excluded that the higher level of hCG in IUI pregnancies is due to an additional vanished implantation. Wider implications of the findings The mechanisms in the embryo and endometrial interplay are far from understood. The present data add to the knowledge regarding this, pointing towards alteration in the implantation window in fresh stimulated cycles. Studies following the more detailed hCG rise are needed for further elucidating how ART affect the early implantation. Trial registration number Journal-nr.: 21019857

P-467 Early (day 1–4) post-treatment serum hCG level changes predict single-dose methotrexate treatment success in tubal ectopic pregnancy

Abstract Study question How effective is the change between day 1 and 4 post-treatment serum hCG level at predicting single-dose methotrexate treatment success in tubal ectopic pregnancy (tEP)? Summary answer Any fall in day 1–4 serum hCG signifies an 85% (95%CI 76.8–90.6) likelihood of treatment success for tEP managed with single-dose methotrexate. What is known already For those with tEP managed by single-dose methotrexate, current guidelines advocate intervention if day 4–7 hCG fails to fall by ≥ 15%. The trajectory of hCG over day 1–4 has been proposed as an early indicator that predicts treatment success, allowing early reassurance for women. However, almost all prior studies of day 1–4 hCG change have been small or retrospective. Study design, size, duration This was a prospective cohort study of women with tEP (pre-treatment hCG of ≥ 1000 IU/L and ≤5000 IU/L) managed with single-dose methotrexate. The data were derived from a UK multicentre randomised controlled trial of methotrexate and gefitinib versus methotrexate and placebo for treatment of tEP (GEM3). For this analysis, we include data from both treatment arms. Participants/materials, setting, methods Participants were categorised by single-dose methotrexate treatment success or failure. Treatment success for this analysis was defined as complete tEP resolution to serum hCG &amp;lt;30 IU/L following single-dose methotrexate treatment only. Patient characteristics between groups were compared. Changes in day 1–4, 1–7 and 4–7 serum hCG were evaluated as predictors of treatment success through receiver operating characteristic (ROC) curve analysis. Test performance characteristics were calculated for percentage reduction thresholds including optimal thresholds. Main results and the role of chance A total of 322 women with tEP and hCG ≥1000 IU/L and ≤5000 IU/L were treated with single-dose methotrexate. The overall single-dose methotrexate treatment success rate was 59% (n = 189/322). Day 1–4, 1–7 and 4–7 serum hCG change predicted single-dose methotrexate treatment success with ROC area under curve 0.80 (95% CI 0.74–0.85), 0.86 (95% CI 0.81–0.91) and 0.89 (95% CI 0.85–0.93) respectively. Any fall between day 1 and day 4 hCG predicted single-dose methotrexate treatment success with a positive predictive value of 85%, negative predictive value 57%, sensitivity 58%, specificity 84%, positive likelihood ratio 3.6 and negative likelihood ratio 0.5. A less than 18% rise in day 1–4 serum hCG was identified as an optimal classification threshold and predicted treatment success with 82% positive predictive value, 69% negative predictive value, 79% sensitivity, 74% specificity, positive likelihood ratio 3.0 and negative likelihood ratio 0.3. Limitations, reasons for caution Our findings may be limited by intervention bias resulting from existing guidelines which influences evaluation of hCG changes reliant on day 7 serum hCG levels. Wider implications of the findings This large prospective cohort demonstrates the value of day 1–4 serum hCG changes in predicting single-dose methotrexate treatment success in tEP. We recommend clinicians provide early reassurance to women who have fall or modest (&amp;lt;18%) rise in day 1–4 serum hCG that their treatment will likely be effective. Trial registration number This study is a secondary analysis of the GEM3 trial (ISRCTN Registry ISRCTN67795930).

P-138 Protein concentration of 5 mg/mL in recovery medium for thawed blastocysts is equivalent to 10 mg/mL in post-thaw survival and pregnancy rates

Abstract Study question Are comparable survival and pregnancy rates obtained when recovery media are supplemented with 5mg/mL protein versus customary 10mg/mL protein in Frozen Embryo Transfer cycles? Summary answer Recovery medium containing 5mg/mL protein compared to 10mg/mL yielded similar outcomes in blastocyst survival and pregnancy rates. What is known already Accepted practice has been to culture thawed blastocysts in medium with the same protein concentration as the vitrification media (20% protein) to minimize osmotic stress and optimize survival and implantation rates. The potential to acquire similarly positive outcomes with supplementation of only 5 mg protein/ml has been explored and demonstrated, challenging the requirement for high protein levels in recovery media. Study design, size, duration From July through August 2022, a prospective study was conducted in a university laboratory to compare the survival and implantation rates in Frozen Embryo Transfer cycles using vitrified blastocysts recovered in Continuous Single Culture–NX Complete (CSCM-NXC) (5mg/mL Human Serum Albumin (HSA)) or global HP[LW1] (8.8mg/mL HSA and 1.2mg/mL α-and ß-globulins). 50 blastocysts were warmed per condition. Comparative analyses included survival and implantation rates, beta HCG levels, and clinical pregnancy rates by fetal cardiac activity. Participants/materials, setting, methods To evaluate the effect of protein level in recovery medium for vitrified blastocysts, CSCM-NXC and global HP, with 5mg/mL and 10mg/mL total protein, respectively, were compared. Blastocysts were vitrified in Irvine Scientific Vit Kit – Freeze and were warmed in Irvine Scientific Vit Kit – Warm. Minimum recovery time was the same for all blastocysts (30 minutes), and is defined as the time from completion of the warming procedure to the time of embryo transfer. Main results and the role of chance In both study arms, one blastocyst was removed from the data analysis due to embryo transfer cancellation that was not related to embryo survival or suitability for transfer, resulting in N = 49 for each group. Chi-square analysis was performed to assess statistical significance of all compared parameters, and a p value &amp;lt; 0.05 was considered significant. Average patient age (years) for CSCM-NXC was 36.1 and for global HP, 35.0. There was no difference between blastocysts warmed in CSCM-NXC and global HP in average recovery time, i.e., 3.8 and 3.9 hours, respectively, or survival rates, 98.4% and 98.1% respectively. Comparison of patients with blastocysts warmed in CSCM-NXC reflected a trend towards higher outcomes compared to those in global HP with initial positive beta HCG rates of 64.6% vs 56.3% and ongoing clinical pregnancy rates of 52.1% vs 45.8%, respectively. Implantation rates were 56.1% and 57.7% in CSCM-NXC and global HP, respectively. Incidence of monozygotic twinning of 0% in CSM-NXC and 3.3% in Global HP was observed. Limitations, reasons for caution As this is a challenge to accepted formulation of recovery media for vitrified blastocysts, specifically with respect to protein supplementation - a strategy for minimizing osmotic shock and potential damage to warming embryos - more expansive and robust studies must be undertaken to increase the statistical power of this study. Wider implications of the findings As frozen embryo transfers are utilized more routinely in assisted reproductive treatment, the clinical finding that a commercial recovery medium containing of 5mg/mL protein, the standard for routine embryo culture, can support the recovery of vitrified embryos and lead to successful pregnancy could increase the efficiency in the laboratory. Trial registration number na

O-141 Combination of gefitinib and methotrexate to treat tubal ectopic pregnancy (GEM3): a multicentre, randomised, double-blind, placebo-controlled trial

Abstract Study question Is a combination of parenteral methotrexate and oral gefitinib more effective than methotrexate alone in the treatment of tubal ectopic pregnancy? Summary answer In women with a tubal ectopic pregnancy, adding oral gefitinib to parental methotrexate does not offer clinical benefit over methotrexate and increases minor adverse reactions. What is known already Current treatment of tubal ectopic pregnancies is with methotrexate or surgery. Methotrexate treatment fails in ∼30% of women and they require rescue surgery. At the time of the design of the trial, it had been shown in preclinical studies that tubal implantation sites express high levels of epidermal growth factor receptor (EGFR) and that gefitinib (an EGFR antagonist) augments methotrexate-induced regression of pregnancy-like tissue. There was also evidence from uncontrolled phase I and II trials that raised the possibility that combination methotrexate and gefitinib could be a more effective medical treatment than methotrexate alone to treat stable ectopic pregnancies. Study design, size, duration Between 2nd November 2016 and 6th October 2021, we performed a multicentre, randomised, double-blind, placebo-controlled trial across 50 UK hospitals. Eligible participants were women with a tubal ectopic pregnancy deemed suitable for medical management with methotrexate. Inclusion criteria were: aged 18-50 years; pre-treatment serum hCG level of 1000–5000 IU/L; and either a definite diagnosis of tubal ectopic pregnancy or a clinical judgement of ‘probable’ tubal ectopic pregnancy. Participants/materials, setting, methods Participants were administered a single dose of intramuscular methotrexate (50 mg/m2) and randomised (1:1 ratio) to seven days of additional oral gefitinib (250mg daily) or placebo. The primary outcome, analysed by intention to treat, was surgical intervention to resolve the ectopic pregnancy. Secondary outcomes included time to resolution of ectopic pregnancy and serious adverse events. Main results and the role of chance 328 participants were allocated to methotrexate and gefitinib (n = 165) or methotrexate and placebo (n = 163). Three participants in the placebo group withdrew. Surgical intervention occurred in 30% (50/165) of the gefitinib group and in 29% (47/160) of the placebo group (adjusted risk ratio 1.15, 95% confidence interval [CI] 0.85-1.58; adjusted risk difference -0.01, 95% CI -0.10-0.09; p = 0.37). Without surgical intervention, median time to resolution was 28.0 days in the gefitinib group and 28.0 days in the placebo group (subdistribution hazard ratio 1.03, 95% CI 0.75-1.40). Serious adverse events occurred in 3% (5/165) of the gefitinib group and in 4% (6/162) of the placebo group. Diarrhoea and rash were more common in the gefitinib group. Limitations, reasons for caution Limitations of the trial include the fact that we only tested one dose regimen. It is possible gefitinib may be effective if a different protocol were used, such as a longer period of administration. Also, we did not carry out pharmacodynamic studies to determine optimal drug bioavailability. Wider implications of the findings Our results show that the addition of gefitinib to standard medical management with methotrexate to treat tubal ectopic pregnancy is not clinically effective as it does not reduce subsequent surgical intervention and is associated with higher rates of reported symptoms than placebo. Trial registration number ISRCTN67795930

What Should be the Thyroid Stimulating Hormone Cut-off Level in the First-Trimester Screening? A prospective Cohort Study and Mini Review of The Guidelines

Aim: Thyroid Stimulating Hormone (TSH) and Human Chorionic Gonadotropin (hCG) are glycoproteins that the alpha subunit common. As a consequence of this, hCG has intrinsic thyrotropic activity. Because of high levels of hCG, maternal serum thyrotropin level is seen to decrease in 80% of pregnancies. Novel investigation and several commission opinions recommend that the TSH cut-off value should be 2.5 mU/L in the first-trimester thyroid function screening.. We aimed to evaluate the contributions of the maternal serum TSH cut-off value of 2.5 mU/L to the development of maternal and fetal complications. Materials and Methods: We constructed the study with pregnancies in their first 12 weeks and planned to do only observation prospectively. We excluded pregnant women with systemic disease and any history of thyroid surgery or thyroid pathology. According to the TSH level, a case group(TSH level &amp;gt;2.5 mU/L) and a control group (TSH level&amp;lt;2.5 mU/L) were created through the pregnants with normal Thyroxine (T4) levels. The cohort group were divided into four subgroups according to whether they were anti-thyroid peroxidase (anti-TPO) positive or not. We observed the fetomaternal outcomes like pregnancy loss, hyperemesis gravidarum, hypertensive disorders, gestational diabetes, prelabour rupture of membranes, placental abruption, with routine prenatal visits until delivery; also delivery style, birth weight, shoulder dystocia, newborn intensive care needs, and postpartum hemorrhage were recorded. Results: The incidence of miscarriage in the subgroup with TSH &amp;gt;2.5 mU/L and anti-TPO (+) was significantly higher than in those with TSH &amp;lt;2.5 mU/L and anti-TPO (+) (p&amp;lt;0.05). All groups had no significant difference in other maternal or fetal/neonatal complications. Conclusion: If only the population-based nomograms are created, we may advise maternal serum TSH level as &amp;lt;2.5 mU/L for first-trimester screening. Single or multiple pregnancy status, gestational age, and the presence of thyroid peroxidase antibodies should also be taken into account when creating these nomograms.