Basal LH Levels Research Articles

Revising LH cut-off for the diagnosis of central precocious puberty via triptorelin stimulation assay.

Precocious puberty (PP) in girls is defined by thelarche before age 8. The diagnostic gold standard is an increased LH level following gonadotropin-releasing hormone (GnRH) stimulation. Alternatively, GnRH analogues like triptorelin can be used, though their interpretation varies. Since 2000, we have used a triptorelin-induced LH cut-off of 15 IU/L, 4 h post-stimulus. However, many girls showed LH values below this threshold despite evident pubertal progression. To establish a new LH threshold post-triptorelin stimulation for earlier diagnosis of central precocious puberty (CPP) in girls showing pubertal progression and to evaluate additional parameters for diagnostic accuracy. We enrolled 186 girls with thelarche onset between ages 1-8 and a GnRH analogue assay performed between 2015-2019 without signs of axis activation. Within this cohort, 62 patients repeated the triptorelin test due to rapid pubertal progression. The assay involved administering 100 mcg/m² of triptorelin and measuring LH, FSH, and estradiol levels before and four hours post-injection. Patients with axis activation at the second test had significantly higher post-stimulus LH levels at the first test compared to those below 15 IU/L. They also had higher basal LH levels, elevated LH/FSH ratio, and increased growth velocity. Statistical analysis identified a new post-stimulus LH threshold of 5 IU/L. We propose a LH value of 5 IU/L after triptorelin administration as a new threshold for early CPP diagnosis. While the LH/FSH ratio and growth velocity are associated with axis activation, they did not significantly enhance diagnostic accuracy when combined with the LH value.

Selective depletion of kisspeptin neurons in the hypothalamic arcuate nucleus in early juvenile life reduces pubertal LH secretion and delays puberty onset in mice.

Puberty is the critical developmental transition to reproductive capability driven by the activation of gonadotropin-releasing hormone (GnRH) neurons. The complex neural mechanisms underlying pubertal activation of GnRH secretion still remain unknown, yet likely include kisspeptin neurons. However, kisspeptin neurons reside in several hypothalamic areas and the specific kisspeptin population timing pubertal onset remains undetermined. To investigate this, we strategically capitalized on the differential ontological expression of the Kiss1 gene in different hypothalamic nuclei to selectively ablate just arcuate kisspeptin neurons (aka KNDy neurons) during the early juvenile period, well before puberty, while sparing RP3V kisspeptin neurons. Both male and female transgenic mice with a majority of their KNDy neurons ablated (KNDyABL) by diphtheria toxin treatment in juvenile life demonstrated significantly delayed puberty onset and lower peripubertal LH secretion than controls. In adulthood, KNDyABL mice demonstrated normal invivo LH pulse frequency with lower basal and peak LH levels, suggesting that only a small subset of KNDy neurons is sufficient for normal GnRH pulse timing but more KNDy cells are needed to secrete normal LH concentrations. Unlike prior KNDy ablation studies in rats, there was no alteration in the occurrence or magnitude of estradiol-induced LH surges in KNDyABL female mice, indicating that a complete KNDy neuronal population is not essential for normal LH surge generation. This study teases apart the contributions of different kisspeptin neural populations to the control of puberty onset, demonstrating that a majority of KNDy neurons in the arcuate nucleus are necessary for the proper timing of puberty in both sexes.

8428 Rare Variants in the MECP2 Gene in Two Boys with Central Precocious Puberty

Abstract Disclosure: A.P. Canton: None. J.B. Mebarak: None. M. Magnuson: None. S. Roberts: None. N. Mauras: None. M. Benson: None. S. Witchel: None. R.S. Carroll: None. A. Latronico: None. U.B. Kaiser: None. A. Abreu: None. Introduction: Central precocious puberty (CPP) occurs more frequently in girls and is usually labelled as idiopathic; however, in boys organic causes are more frequent. Identification of imprinted genes causing CPP have revealed epigenetic mechanisms underlying puberty. MECP2, an X-linked gene, encodes a methylated DNA reader protein with a role in gene transcription. MECP2 loss-of-function mutations usually cause Rett syndrome, a severe neurodevelopmental disorder that may be associated with early pubertal development. Recently, rare variants in MECP2 have been recognized in girls with sporadic CPP with or without mild neurodevelopmental disorders. In our cohort of 78 patients with idiopathic CPP, no MECP2 mutations were identified in 73 girls. In this study, five boys with CPP were evaluated for potential MECP2 sequence variants. Methods and Results: Five boys with CPP were screened for MECP2 sequence variants using Sanger sequencing. At the time of CPP diagnosis, they had median (interquartile range) chronological age 9.2 yr (4.3), bone age advancement 2.1 yr (3.5), height SDS 2.3 (1.5), basal LH levels 1.6 IU/L (0.9), and testosterone levels 370 ng/dL (570). Organic causes of CPP were excluded. No MKRN3 or DLK1 mutations were identified. Familial segregation analysis was performed when appropriate. We identified a hemizygous MECP2 variant in a boy (Patient 1) who presented at age 2.7 yr with sporadic CPP. In addition, he had speech delay and behavioral changes, defined as autism spectrum disorder by neuropsychological assessment. He harbored an extremely rare (gnomAD AF=0.000004956) missense variant (p.Val312Ile) in exon 3 of MECP2, encoding the transcriptional repression domain, critical for protein function. The p.Val312Ile mutation was classified as likely pathogenic (ACMG criteria) with a potential association with the phenotype. Another hemizygous MECP2 variant was identified in a boy (Patient 2) who presented at age 8.0 yr with sporadic CPP; he had no neurodevelopmental conditions. He harbored a rare (gnomAD AF=0.00008) missense variant (p.Arg366Cys) in exon 3, encoding the C-terminal domain, classified as a variant of uncertain significance (ACMG criteria). Familial segregation analysis revealed that both boys inherited the MECP2 variants from their unaffected mothers, consistent with a pattern of clinical variability described in women with defects in X-linked genes.Conclusions: We identified rare MECP2 variants in two boys with sporadic CPP without classic features of Rett syndrome, expanding the phenotype of patients with MECP2 mutations, as described in girls. The MECP2 p.Val312Ile variant was likely pathogenic, whereas the functional significance of the p.Arg366Cys variant in still indeterminate. These findings provide additional evidence for a role of MECP2, an epigenetic factor, in the hypothalamic control of pubertal timing. Presentation: 6/1/2024

7381 Overexpression of Makorin Ring Finger Protein 3 in the Arcuate Nucleus of Postpubertal Female Mice Impacts Neurokinin B and Kisspeptin Co-Expressing Neurons

Abstract Disclosure: S.A. Roberts: None. A. Fontes: None. S. Blau: None. M. Magnuson: None. K.M. McKnight: None. M. Sena-Esteves: None. R.S. Carroll: None. U.B. Kaiser: None. Background: Makorin ring finger protein 3 (MKRN3), an upstream inhibitor of GnRH release, is an important regulator of the age of menarche. Loss-of-function mutations in MKRN3 are the most common genetic etiology of central precocious puberty. However, its potential role in delayed puberty or hypogonadism is not well defined. In mice, expression of Mkrn3 in the hypothalamic arcuate nucleus (ARC), where kisspeptin, neurokinin B and dynorphin (KNDy) co-expressing neurons are located, is high early in life and declines before pubertal onset. Neonatal hypothalamic overexpression of Mkrn3 leads to delayed puberty in female mice and decreased neurokinin B (NKB) and kisspeptin protein levels in the ARC peripubertally. Conversely, Mkrn3-deficient peripubertal mice exhibit increased NKB protein levels in the ARC. We hypothesized that Mkrn3 overexpression in the ARC of postpubertal female mice would result in suppression of the HPG axis, manifested as hypogonadotropic hypogonadism. Methods and Results: Twelve week old adult wild type female mice were injected stereotaxically into the ARC with a novel adeno-associated virus overexpressing Mkrn3 (AAV-Mkrn3-IRES-EGFP; denoted AAV-Mkrn3) or a control virus (AAV-EGFP). Mean body weight did not differ between groups post-injection. AAV-Mkrn3 injected mice spent significantly more time (62.7%) in diestrus three to six weeks post-injection compared to controls (41.3%; P<0.01). Eight weeks post-injection, gonad-intact females were injected with an NKB receptor (NK3R) agonist, senktide (5 nmol or 10 nmol ip). AAV-Mkrn3 mice had a decreased peak LH response twenty minutes post-injection, compared to controls (P<0.001) at both doses. Peak LH levels were also reduced in response to senktide (5 nmol ip) in ovariectomized and estradiol capsule replaced (OVX+E2) AAV-Mkrn3 injected females, compared to controls (P< 0.001). In contrast, in both gonad-intact and OVX+E2 female mice, kisspeptin-10 (1 nmol ip) resulted in similar peak serum LH levels in AAV-Mkrn3 injected mice compared to controls. Immunohistochemistry in the ARC of intact and OVX+E2 AAV-Mkrn3 injected females showed significantly decreased NKB protein expression in AAV-Mkrn3 mice compared to controls. In response to OVX, serum LH levels increased similarly between AAV-Mkrn3 and control-injected mice. LH pulsatility in OVX females, assessed over 3 hours, showed that mean basal LH levels, number of pulses, pulse amplitude and area under the curve did not differ significantly between groups. Conclusions: These findings indicate that MKRN3 can exert inhibitory effects on the reproductive axis in female mice beyond the juvenile period, which has implications as a potential therapeutic target. The blunted response to senktide, and decreased NKB protein levels in the setting of Mkrn3 overexpression, further supports the impact of Mkrn3 on the neuropeptides and/or receptors of KNDy neurons. Presentation: 6/2/2024

Pregnancy and perinatal outcomes in pregnancies following frozen embryo transfer (FET) after transcervical resection of adhesions (TCRA): A retrospective cohort study with propensity score matching analysis.

To compare the pregnancy and perinatal outcomes of frozen-thawed embryo transfer (FET) in patients following transcervical resection of adhesions (TCRA) versus patients with normal uterine morphology, and to investigate the factors influencing pregnancy outcomes in patients undergoing FET after TCRA. We retrospectively analyzed FET cycles from September 2014 to September 2023, comparing patients with normal uterine morphology to those with intrauterine adhesions (IUAs) treated with TCRA. Propensity score matching (PSM) adjusted for confounding factors. LASSO regression and multivariate logistic regression identified predictors of outcomes, which were visually represented in nomograms. Model performance was assessed using calibration curves, ROC curves, and DCA, with bootstrap method for internal validation. Post-PSM analysis showed higher live birth rates in patients with normal uterine morphology after clinical pregnancy (75.1% vs. 61.7%, P < 0.001). No significant differences were noted in clinical pregnancy rates and perinatal outcomes between the groups. Factors influencing clinical pregnancy in FET after TCRA included basal progesterone levels, endometrial thickness, parity, infertility cause, embryo stage at transfer, number and quality of embryos transferred, IUA severity, and TCRA surgical procedures. Body mass index, basal LH levels, and day 14 HCG levels post-embryo transfer were determinants of live birth outcome. FET cycles following TCRA showed a lower rate of successful live births, but TCRA did not increase adverse perinatal outcome risks. Our study introduces an innovative predictive model for clinical pregnancy and live birth outcomes in patients undergoing FET following TCRA, addressing a significant void in existing research.

Should non-invasive prenatal testing be recommended for patients who achieve pregnancy with PGT?

ObjectiveTo determine whether non-invasive prenatal testing is an alternative testing option to preimplantation genetic testing (PGT) in pregnant patients.MethodsThis was a retrospective study of the clinical outcomes of patients who underwent PGT and invasive or non-invasive pregnancy testing after euploid blastocyst transfer at our IVF centre between January 2017 and December 2022.ResultsIn total, 321 patients were enrolled in this study, 138 (43.0%) received invasive pregnancy testing, and 183 (57.0%) patients underwent non-invasive testing. The mean age of the patients in Group 2 was higher than that of the patients in Group 1 (35.64 ± 4.74 vs. 31.04 ± 4.15 years, P < 0.001). The basal LH and AMH levels were higher in Group 1 than in Group 2 (4.30 ± 2.68 vs. 3.40 ± 1.88, P = 0.003; 5.55 ± 11.22 vs. 4.09 ± 3.55, P = 0.012), but the clinical outcomes were not significantly different. Furthermore, the clinical outcomes of patients undergoing invasive testing were similar to those of patients undergoing non-invasive testing with the same PGT indication.ConclusionOur results suggest that non-invasive pregnancy testing is a suitable alternative option for detecting the foetal chromosomal status in a PGT cycle. However, the usefulness of non-invasive testing in PGT-M patients is still limited.

Treatment of D-galactose-induced rat polycystic ovarian condition using Lepidium sativum and secondary antibodies.

There is still much to be discovered regarding the etiopathogenesis and management of polycystic ovarian syndrome (PCOS). Four groups of female Wister-Albino rats were established, each with a normal estrous cycle: control, D ( + ) galactose (D-galactose), Lepidium sativum (L. sativum), and prepared secondary antibody (Ab2). Serum samples were collected, and histopathological examination was performed on ovaries and spleen tissues. Immunoreactive anti-ovarian antibody (AOA) quantities were determined using a modified antigen-based ELISA procedure. ELISA assay kits were used to quantify FSH, LH, and estradiol 17 β concentrations. The study found that AOA concentration in undiluted samples was significantly higher in the second and fourth weeks after PCOS induction by D-galactose (p < 0.001). However, antibody index% and titer elevated in the D-galactose group. L. sativum's late efficacy was observed in the fourth week, while the concentration of undiluted samples in the D-galactose + Ab2 group lowered (p < 0.001). Higher basal FSH and LH levels and lower estrogen levels are associated with PCOS development. L. sativum's immunomodulatory properties may contribute to this association. Estradiol-17ß concentrations increased in D-galactose + L. sativum and D-galactose + Ab2 groups, respectively. Careful extrapolation of experimental models is crucial for clinical applications, as technical advancements make Ab2 production easier. Further study is needed to fully understand its potential in immunotherapy.

SAT608 Gonadotrope-specific Deletion Of Icer Results In Increased Lh Pulsatility But Reduced Fsh Levels In Mice

Abstract Disclosure: N.P. Boris: None. S.A. Pereira: None. G.A. Stamatiades: None. K.N. Hausken: None. H. Kim: None. R.S. Carroll: None. U.B. Kaiser: None. Background: The hypothalamic-pituitary-gonadal (HPG) axis is regulated by pulsatile secretion of hypothalamic GnRH, which acts on anterior pituitary gonadotropes to control synthesis and secretion of LH and FSH. High and low GnRH pulse frequencies preferentially stimulate Lhb and Fshb expression, respectively, through mechanisms that are not completely understood. The mouse Fshb gene promoter contains a half-AP-1/half-cAMP response element (CRE) motif; we have shown previously that this element is bound by CRE-binding protein (Creb) in vitro to increase expression in response to low GnRH pulse frequencies, but is antagonized by inducible cAMP early repressor (Icer) at higher GnRH pulse frequencies, resulting in a lower level of Fshb transcription. In this study, we aimed to identify the role of Icer in gonadotropin regulation in vivo in mice. We hypothesized that gonadotrope-specific deletion of Icer may influence pubertal timing and fertility due to an increase in Fshb expression. Methods and Results: Gonadotrope-specific deletion of Icer (GnrhrGRIC/+/Icerfl/fl; Icer KO) and control (Icerfl/fl) mice were generated, and pubertal and reproductive phenotypes were evaluated. There was no difference in body weight or in pubertal timing, evaluated by age of preputial separation in males and by vaginal opening and first estrus in females. In post-pubertal male mice, neither basal LH and FSH levels nor the LH or FSH response to acute GnRH stimulation were significantly different in Icer KO mice, compared to littermate Icerfl/fl controls. Male Icer KO mice 14 days post-gonadectomy (GDX) had significantly more LH pulses within a three-hour period than controls (Icer KO 5.83 ± 0.4 pulses vs Icerfl/fl 4.66 ± 0.3 pulses; p=0.049), but no differences in basal LH or LH pulse amplitude. Interestingly, serum FSH levels were significantly lower in Icer KO mice compared to Icerfl/fl controls 2 days post-GDX (Icer KO 20.0 ± 1.0 vs Icerfl/fl 26.0 ± 0.8; p=0.003) and 7 days post-GDX (Icer KO 18.0 ± 0.4 vs Icerfl/fl 22.5 ± 0.8, p=0.002). There were no significant differences in pituitary Fshb, Lhb, or Gnrhr mRNA levels between Icer KO and Icerfl/fl control mice 14 days post-GDX. Conclusions: Deletion of Icer from pituitary gonadotropes in mice did not affect body weight or timing of pubertal onset. However, in GDX male mice, LH pulse frequency was increased, whereas the post-GDX increase in serum FSH levels was blunted, in Icer KO mice compared to controls. These findings support a role for Icer in both LH and FSH regulation in vivo and highlight the importance of extending in vitro observations to in vivo models. Presentation: Saturday, June 17, 2023

LH level on the antagonist administration day as a predictor of the reproductive outcomes in women with normal ovarian function.

The addition of antagonists is mainly based on estrogen level and follicle size, while LH level has not received sufficient attention.In this study, LH Level on the antagonist administration day was used as the main research objective to explore its relationship with laboratory indicators and pregnancy outcomes. We enrolled 854 patients with normal ovarian function undergoing in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) between May 2021 to May 2022 at the Reproductive Center of Shandong University of Traditional Chinese Medicine.We used the quartile method to group LH levels on the antagonist administration day. There were four groups: Q1 (0.53IU/L≤LH ≤ 1.89IU/L); Q2 (1.89IU/L<LH ≤ 3.01IU/L); Q3 (3.01IU/L<LH≤ 5.29 IU/L); Q4 (5.29IU/L<LH ≤ 8.72IU/L). A total of 452 fresh embryo transplantation cycles and 1726 Frozen embryo transplantation cycles were carried out. There were significant differences among the four groups in terms of total Gn dosage, E2, P and LH on trigger day, number of retrieved oocytes, number of 2PN embryos, number of blastocysts, Number of ET and fresh ETR.There is a significant correlation between LH on antagonist administration day and Basal LH Level,LH on trigger day,number of oocytes retrieved,number of 2PN embryos,number of blastocysts, number of ET.Using Fresh ETR,Fresh CPR,OHSS and Cumulative CPR as the criterion respectively, the optimal cut-off value for evaluating LH on antagonist administration day was 4.18IU/L,3.99IU/L,4.63IU/L,4.66IU/L. There was a significant positive correlation between LH on the antagonist administration day and number of oocytes retrieved,number of 2PN embryos,number of blastocysts.LH on the antagonist administration day could predict Fresh CPR,OHSS and Cumulative CPR to some extent.

Effect of basal luteinizing hormone (bLH) level on in vitro fertilization/intra-cytoplasmic injections (IVF/ICSI) outcomes in polycystic ovarian syndrome (PCOS) patients

ObjectiveTo evaluate the effect of basal luteinizing hormone (bLH) levels on In Vitro Fertilization/Intra-Cytoplasmic Injections (IVF/ICSI) outcomes in polycystic ovary syndrome (PCOS).MethodsA total of 256 PCOS patients who underwent IVF/ICSI treatment in our center from January 2018 to January 2022 were analyzed retrospectively. The patients were based on the third quartile (12.455) of the basal LH value was taken as the cut-off value and was divided into high and low LH group: high LH group (LH ≥ 12.455 IU / L) and low LH group (LH < 12.455 IU / L) and the OC group was pretreated with oral contraceptives. The outcomes in ovulation induction and embryo transfer cycles of the three groups were then compared. In addition, factors influencing the number of good quality embryos and the early onset LH peak were analyzed.ResultsAges, infertility duration, body mass index (BMI), and basal follicle-stimulating hormone (FSH), and progesterone (P), testosterone (T) levels were not significantly different among the three groups (p > 0.05). However,there were significant differences in basal LH and basal E2 between low LH group and high LH group, and there were significant differences in basal LH between high LH group and OC group (p < 0.05). LH on the antagonist day was significantly different between low LH group and high LH group and between high LH group and OC group (p < 0.05). LH on the hCG (human Chorionic Gonadotropin) day there were significant differences between low LH group and OC group, high LH group and OC group (p < 0.05). The Mode of triggering between the three groups had significant differences between the two groups (p < 0.05). In addition, the number of days from gonadotropin (Gn) initiation to antagonist addition were significantly different among the three groups (p < 0.05). In addition, total Gn doses,the number of oocytes retrieved, the number of Gn days, 2pronucleus (2PN) numbers, number of good quality embryos, and number of high risk OHSS (Ovarian Hyper-stimulation Syndrome), cases with OHSS occurrences were not significantly different among the three groups (p > 0.05). Moreover, the cycle and clinical pregnancy outcomes and the cumulative clinical pregnancy rate and the cumulative live birth rate were not significantly different among the three groups (p > 0.05). LH levels on the day of antagonist addition affected the number of good-quality embryos (B < 0, p < 0.05). However, LH levels on the day antagonist was added were not significantly correlated with basal LH levels (Pearson correlation coefficient = 0.259), the ROC curve was constructed for the logistic prediction model of the early onset LH peak, and the AUC value was 0.747, indicating that the logistic combined model we constructed had a good ability to predict the early onset LH peak.ConclusionBasal LH levels do not affect the pregnancy outcomes in PCOS patients after antagonist protocols. Besides, LH levels on the day of antagonist addition affect the number of good quality embryos for PCOS patients undergoing IVF /ICSI.

Analysis of serum insulin-like growth factor-1, fibroblast growth factor 23, and Klotho levels in girls with rapidly progressive central precocious puberty

To study the levels of serum insulin-like growth factor 1 (IGF-1), fibroblast growth factor 23 (FGF23), and Klotho, and to study their relationship with girls with rapidly progressive central precocious puberty (RP-CPP). This is a cross-sectional study on the progression rate of central precocious puberty in girls, who complained of breast development before the age of 8 years and were followed between June 2021 and June 2022. At the same time, 28 healthy girls less than 8 years old who had not started puberty were recruited as the control group. The physical examination and laboratory evaluation of each group was completed. Only patients with CPP received pelvic ultrasound examination and bone age test. Bone age index (BAI), basal LH levels (BLH), basal LH levels/basal FSH levels (BFSH), peak LH (PLH)/peak FSH (PFSH), IGF-1, Klotho, FGF23, and ovarian volume in the RP-CPP group were higher than those in slowly progressive CPP (SP-CPP) group. In the RP-CPP group, IGF-1 was correlated with Klotho, FGF23, and BLH; Klotho was correlated with FGF23 and BLH; FGF23 was correlated with BLH.Conclusion: The BLH, FGF23, Klotho, and IGF-1 have a certain correlation with RP-CPP, which may play an important role in the speed of girls’ sexual development.What is Known:• The association between IGF-1 and RP-CPP.What is New:• We found the association between FGF23, Klotho and RP-CPP.

Role of luteinizing hormone urinary levels in the diagnostic and therapeutic management of female central precocious puberty

BackgroundDiagnosing central precocious puberty (CPP) requires an integrated approach based on clinical, biochemical and instrumental data. The diagnostic gold standard is represented by GnRH (gonadotropin-releasing hormone) stimulation test. Some undoubted limitations of this procedure led the international scientific community to look for cheaper and less invasive alternative diagnostic methods, such as luteinizing hormone urinary levels (uLH) measurement. This study aims to define the reliability of urinary LH levels as a biomarker of pubertal development, both concerning the initial diagnostic management and the monitoring of patients with central precocious puberty undergoing therapy with GnRH analogues. Furthermore, the study plans to detect the potential association between LH peak serum (pLH) and urinary LH in patients undergoing diagnostic tests with GnRH and to identify a possible cut-off of uLH that may be suggestive of ensued successful hormonal stimulation.MethodsThe study includes 130 female patients with suspected precocious puberty or in follow-up during suppressive therapy. After the collection of the informed consent, the patients underwent clinical evaluation, auxological assessment, and hormone assays (basal levels of LH, FSH, and oestradiol; GnRH stimulating test in patients with suspected precocious puberty; urinary LH assay on the first-morning urine sample, collected after waking up).ResultsTwo uLH cut-off values have been identified: the first of 0.25 UI/L [C.I. 95% 0.23–0.27], able to distinguish between pubertal and pre-pubertal patients, the second of 0.45 UI/L [C.I. 95% 0,20 − 0,70] suggestive of occurred hormonal stimulation in patients with diagnosis of CPP at GnRH test. All 30 patients with CPP in follow-up during suppressive therapy presented uLH values ≤ 0.45 IU/L (pU < 0.05), and uLH collected in prepubertal group control.ConclusionsuLH assays on the first morning urine specimen could be considered a low-cost and minimally invasive tool for precocious puberty diagnosing and monitoring, making possible to be easily performed even by a general pediatrician. Thus, this could help referring only selected patients to pediatric endocrinologists. After an appropriate validation, this approach could reasonably reduce hospital attendance and costs of performing more invasive procedures, with a more significant emotional impact on the pediatric patient.

An Algorithm to Predict the Lack of Pregnancy after Intrauterine Insemination in Infertile Patients.

Increasing intrauterine insemination (IUI) success rates is essential to improve the quality of care for infertile couples. Additionally, straight referral of couples with less probability of achieving a pregnancy through IUI to more complex methods such as in vitro fertilization is important to reduce costs and the time to pregnancy. The aim of the present study is to prospectively evaluate the threshold values for different parameters related to success in intrauterine insemination in order to provide better reproductive counseling to infertile couples, moreover, to generate an algorithm based on male and female parameters to predict whether the couple is suitable for achieving pregnancy using IUI. For that, one hundred ninety-seven infertile couples undergoing 409 consecutive cycles of intrauterine insemination during a two-year period were included. The first year served as a definition of the parameters and thresholds related to pregnancy achievement, while the second year was used to validate the consistency of these parameters. Subsequently, those parameters that remained consistent throughout two years were included in a generalized estimating equation model (GEE) to determine their significance in predicting pregnancy achievement. Parameters significantly associated with the lack of pregnancy through IUI and included in the GEE were (p < 0.05): (i) male age > 41 years; (ii) ejaculate sperm count < 51.79 x 106 sperm; (iii) swim-up alkaline Comet > 59%; (iv) female body mass index > 45 kg/m2; (v) duration of infertility (>84 months), and (vi) basal LH levels > 27.28 mUI/mL. The application of these limits could provide a pregnancy prognosis to couples before undergoing intrauterine insemination, therefore avoiding it in couples with low chances of success. The retrospective application of these parameters to the same cohort of patients would have increased the pregnancy rate by up to 30%.

Does Baseline Serum Androgen Levels Have an Impact on Ovulation Induction Cycle Outcomes by Using Clomifene Citrate Among Infertile Women with Polycystic Ovary Syndrome? A Retrospective Cohort Study

OBJECTIVE: Hyperandrogenism is one of the diagnostic criteria for polycystic ovary syndrome. Women with polycystic ovary syndrome suffer from infertility due to anovulation. Hyperandrogenism results in follicular arrest at the antral stage during folliculogenesis. baseline intrinsic hyperandrogenism affects the success of ovulation induction or not. The relationship between baseline serum androgen levels and ovulation induction cycle outcomes by using clomifene citrate among infertile women with polycystic ovary syndrome has not been investigated thoroughly. STUDY DESIGN: Ovulation induction cycle outcomes of 35 infertile women diagnosed with polycystic ovary syndrome according to Rotterdam criteria who have received 50-100 mg/day clomifene citrate have been evaluated retrospectively. Menstrual cycle day 2-5 serum levels for gonadotropins, androgens, metabolic parameters, and ovulation induction cycle outcomes have been compared between women who have and have not achieved clinical pregnancy following treatment. RESULTS: Serum basal follicular stimulating hormone, LH, E2, fasting cholesterol, glucose, and HOMA-IR levels were comparable between these two groups of patients. Unlike other serum androgens, baseline serum-free testosterone level is significantly lower for patients who have achieved clinical pregnancy following ovulation induction with clomifene citrate. The baseline serum cut-off level for free testosterone to predict clinical pregnancy was 1.94 pg/ml with 75% sensitivity and 67% specificity rates. CONCLUSION: Lower or higher levels of androgenic milieu within the ovaries result in defective folliculogenesis and ovulation failure. Increased serum levels of free testosterone which is a proxy for ovarian androgen production might be a detrimental factor for clinical pregnancy rates of women with polycystic ovary syndrome by impairing proper folliculogenesis.

State of sexual development in children with congenital epidermolysis bullosa

Purpose of the study. Assess the state of sexual development in children with CEB.Methods and materials. The study included 50 children aged 8.11 to 17.80 years with CEB who were treated at the National Medical Research Centre for Children’s Health (Moscow, Russia) from December 2020 to April 2022. Anthropometric parameters, the stage of sexual development on the Tanner scale, bone age, the level of pituitary (FSH, LH) and sex (estradiol, testosterone) hormones, additional hormonal parameters (cortisol, DHEA-sulfate, 17OHP, ACTH, prolactin, TSH, T3, T4, insulin), ultrasound of the pelvic organs for girls and ultrasound of the scrotum organs in boys, the results of a psychological and pedagogical examination (questionnaire according to S. Bem, ‘drawing of a person’ by K. Machover, D. Wechsler’s test) were assessed in all patients. The data are presented using median (Me), quartiles of 25 and 75% [Q25; Q75] and standard error (SE). Due to the small number of observations, nonparametric statistical criteria were used to assess the significance of the differences in the obtained indicators.Results. According to the results of the study, a statistically significant relationship was established between the occurrence of deviations in sexual development in children with CEB and the clinical form of the disease (p &lt; 0,001). The severity of clinical manifestations of CEB according to the EBDASI scale was significantly higher in children in the group with deviations in sexual development (p = 0,000) compared with the group of children with normal sexual development. Anthropometric indicators of BAZ and HAZ in the group of children with deviations in sexual development were statistically significantly reduced (p = 0.000, p = 0.000, respectively) compared with children with normal sexual development, which means a significantly higher incidence of malnutrition in this cohort of patients. Basal levels of FSH and LH (p = 0,000, p = 0,001, respectively), estradiol and testosterone (p = 0,002, p = 0,000, respectively) were significantly lower in children with abnormalities in sexual development compared with children with normal sexual development. A statistically significant relationship was revealed in the studied groups with the stage of sexual development according to Tanner scale (p = 0,032). Among all patients with CEB (n = 50), delay puberty was diagnosed in 7 patients (14%), while all patients suffered from RDEB and had low basal LH levels, reduced estradiol and testosterone values, which allowed them to be diagnosed with hypogonadotropic hypogonadism (HH).Conclusions. In patients with CEB with the most severe degree of clinical manifestations, accompanied by decompensated malnutrition of multifactorial genesis, there are deviations in sexual development and upon reaching the age of 13 years, girls and boys develop transient (symptomatic).

OR18-2 Clinical, Hormonal and Genetic Characterization of Familial Central Precocious Puberty

Abstract Context: Familial central precocious puberty (CPP) is a prevalent form (about 27.5%) of precocious puberty. Loss-of-function mutations in two maternally imprinted genes, MKRN3 and DLK1, were identified in families with CPP showing dominant autosomal inheritance with paternal transmission. Maternally transmitted CPP was previously demonstrated in up to 60% of families using pedigree analysis, however, no definitive genetic abnormality has been identified so far. Objectives To estimate the prevalence of familial cases in a multiethnic cohort with CPP. To characterize the genetic basis and the mode of inheritance of the affected families. To compare clinical and hormonal features of patients with familial CPP due to different modes of transmission. Patients and Methods Clinical and hormonal data were obtained from medical registries of 495 patients with CPP and no brain MRI alterations. Familial CPP was defined as the presence of one or more close relatives with CPP or precocious menarche (≤9 yr). Sanger sequencing of MKRN3 and DLK1 was performed in 427 index cases. Targeted gene panel sequencing was performed in 79 cases, while whole exome sequencing was performed in 101 cases from 36 families Results Among 495 index cases, 159 had familial CPP (31%). The mode of transmission of CPP was identified as paternal in 58 (35%), maternal in 59 (38%), indeterminate in 34 (22%), and both maternal and paternal transmission in 8 (5%). Most families with paternal or maternal transmission had 2 generations known to be affected (51 and 77%, respectively). Notably, 67% of index cases with maternally transmitted CPP had their mother affected. In girls with CPP, the median age of thelarche was 6.5 yr in the paternally transmitted group, 6.9 yr in the maternally transmitted group, and 7.3 yr in the indeterminate group (p= 0.547). Median bone age advancement was 2.1, 2.3 and 1.2 yr, respectively (p= 0.013). Basal LH levels were higher in girls with paternally transmitted CPP (p= 0.047). Among those with paternally transmitted CPP, MKRN3 and DLK1 mutations were identified in, respectively, 63.8% and 10.3% of the families. Conclusions A significant prevalence (31%) of familial CPP was demonstrated in a multiethnic cohort. Maternally transmitted CPP represented the most frequent form of familial CPP (38%). MKRN3 loss-of-function mutations were responsible for most paternally transmitted CPP cases (63.8%), followed by DLK1 loss-of-function mutations (10.3%). Presentation: Monday, June 13, 2022 11:15 a.m. - 11:30 a.m.

Predictive Value of Basal Serum Progesterone for Successful IVF in Endometriosis Patients: The Need for a Personalized Approach.

The data regarding the role of progesterone (P4) in reproductive events of endometriosis patients are limited. This prospective study aimed to examine the predictive value of basal P4 serum levels for successful in vitro fertilization (IVF) in patients with primary infertility and endometriosis. The study included 73 patients divided according to endometriosis treatment (surgery vs. control—no treatment). The general data, basal hormonal status, and pregnancy rates were determined for every patient. Clinical pregnancy was achieved in 40.3% of patients, and more often in patients treated for endometriosis before IVF. The regression analysis showed that higher basal P4 serum levels were associated with achieving pregnancy through IVF. When regression was adjusted for the patient and IVF characteristics, higher basal P4 serum levels were associated with pregnancy achievement in both groups of women, along with the basal serum levels of FSH, LH, and AMH; EFI score; and stimulation protocol. The ROC analysis showed that the basal P4 serum level for successful IVF should be ≥0.7ng/mL. The basal P4 serum level cut-off for IVF success in endometriosis patients was determined for the first time. Constructed models for IVF success prediction emphasize the importance of determining the basal P4 serum levels for the personalized treatment of endometriosis-related infertility.

Does Serum LH Level Influence IVF Outcomes in Women with PCOS Undergoing GnRH-Antagonist Stimulation: A Novel Indicator.

Objective: To explore the influence of LH levels on the IVF/ICSI outcomes in women with PCOSundergoing GnRH-antagonist stimulation protocol. Methods: A total of 142 IVF/ICSI patients in which the females were diagnosed with PCOS and underwent GnRH-antagonist protocol for ovarian stimulation were enrolled. Patients were divided into three groups based on basal LH (bLH) level, LH level on trigger day (hLH), and the ratio of hLH/bLH. The LH levels detected on different days in the stimulation cycle as well as their relationships with the IVF/ICSI outcomes were investigated. The main outcomes we observed were the number of oocytes retrieved, the cumulative chemical pregnancy rate, clinical pregnancy rate, and live birth rate. Other factors included the number of normally fertilized oocytes (2PN), top-quality embryo rate, and total Gn dose. Results: There was no significant difference in the included outcomes and baseline characteristics among different groups based on bLH levels. When patients were grouped according to hLH levels (≤2 mIU/mL, 2–5 mIU/mL and ≥5 mIU/mL), we found decreased levels of basal FSH and LH in the group of hLH ≤ 2 mIU/mL than the other two groups. Then the ratio of hLH/bLH was calculated for each patient. Patients with hLH/bLH ≥ 1 had a higher top-quality embryo rate than those with hLH/bLH between 0.5 and 1.0. Nevertheless, the cumulative clinical pregnancy rate was significantly higher in the hLH/bLH ≤ 0.5 group than in the other two groups. Conclusions: The study proposed the hLH/bLH ratio as a potential in predicting the influence of LH level on the embryo development potential as well as pregnancy outcomes in women with PCOS undergoing GnRH-antagonist stimulation cycles.

Correlation Between BMI And LH Level and LH/FSH Ratio-A Cross-Sectional Study

Background: Polycystic ovarian syndrome is the most common reproductive endocrinopathy of women during their childbearing age. Elevated LH/FSH ratio, abnormally high androgen level and relatively high endogenous estrogen production are indicative of PCOS. The diagnosis of polycystic ovarian syndrome remains controversial with some considering transvaginal sonography with the combination of clinical criteria for diagnosis. In this study, we tried to determine whether there is any impact of BMI on LH or LH/FSH ratio in PCOS for better management in the future.&#x0D; Materials and Methods: This is a cross-sectional study. Those clinically presented as obese, hirsute, and with menstrual disturbances were non-smokers and had not been on any medications for the last three months before the study. Age groups ˂20 and ˃40 years, non-PCOS with infertility, and nonobese patients were excluded from the study.&#x0D; Results: Among the total 58 patients, BMI in the range of 25-28 kg/m 2 was found in the highest frequency in 44 cases (75.9%), high LH level group in 33 cases (56.9%). LH/FSH ratio ˃2 found in 27 cases (46.66%). The correlation between BMI and LH/FSH ratio showed the highest frequency in 21 cases in the BMI group (25-28kg/m 2 ) (p ˂0.009). Random LH level amplitude was found higher in low BMI (25-28kg/m 2 ) than lower in higher BMI (28-34kg/m 2 ) (p ˂0.003).&#x0D; Conclusion:Assessments of basal LH levels and the LH/FSH ratio in hyperandrogenic anovulatory women would be clinically meaningful when BMI is taken into account.&#x0D; TAJ 2022; 35: No-1: 71-76

P-566 Clinical parameters that predict a premature LH rise in patients undergoing ovarian stimulation for IVF

Abstract Study question What are the clinical parameters that can predict a premature LH rise or LH surge in patients undergoing ovarian stimulation for IVF? Summary answer The parameters predicting an early LH rise are: Age, BMI, day 3 LH levels, LH on day of antagonist administration, and number of stimulation days. What is known already Normal reproductive function and fertility require precise regulation of FSH and LH secretion. In IVF stimulation cycles some patients will have an early LH rise even while being treated with a GnRH antagonist. Previous studies have shown a possible link of LH rise with ovarian response to gonadotropins. Nevertheless, little is known of which clinical parameters can predict this LH rise; an occurrence that may necessitate cycle cancellation. Early prediction of premature LH rise will enable us to foresee this unwanted hypophyseal event, and in turn may lead to early antagonist administration and/or early triggering for oocyte retrieval. Study design, size, duration A retrospective study enrolling 382 patients who underwent IVF treatment at Rambam Medical Center (2015 - 2021): age 34.1±6.7 years; BMI 25.0±5.1 kg/m²; AFC 11.2±7.1; day 3 FSH 7.2±2.9U/L; day 3 LH 6.4±4.3U/L; eggs retrieved 8.0±6.1; fertilizations 4.7±4.2. Patients were stimulated with rFSH (Gonal F)/rFSH+LH (Pergoveris)/ highly purified urinary FSH+hCG(Menopur); GnRH antagonist (Cetrotide/ Orgalutran) was administered using a flexible protocol; triggering of ovulation was performed using GnRHagonist and/or hCG, based on estradiol levels. Participants/materials, setting, methods Clinical parameters included from chart data were: age; BMI; estradiol, progesterone, LH and FSH levels at day3/ day of antagonist administration/ and last day before triggering; total stimulation days and total FSH dose. In order to establish a model to predict premature LH rise we performed multiple linear regression using ANOVA with a confidence interval of 0.95 considered significant. A machine learning based model was used to test the accuracy of our model using Python. Main results and the role of chance An LH rise was calculated as the difference between pre-trigger- and basal LH levels. For the total group of patients this LH rise was significantly predicted by patient age; BMI; LH levels at day 3 and LH at the day of antagonist administration; and total stimulation days (R = 0.742, R²=0.55, p &amp;lt; 0.001). Importantly, when analyzing the data of specific age groups, the prediction of the model was strongest in the young patients (age 25-30: R = 0.94, R²=0.88, p &amp;lt; 0.001) and weakest in the older patients (age &amp;gt; 41: R = 0.48,R²=0.23, p = 0.003). Accordingly, using a machine learning model in which a Python based algorithm is trained by analyzing 80% of the data, we were able to successfully predict the remaining 20%: mean squared error of 4.32 and 82.3 in the young- and old patient groups respectively. Interestingly, 21 patients demonstrated a “full LH surge” as defined by a more than two fold increase in LH levels. These patients were also predicted by a similar model (LH levels at day 3 and at the day of antagonist administration, BMI and total FSH dose; R = 0.89, R²=0.80, p = 0.015). Limitations, reasons for caution This is a retrospective study and its conclusions need to be validated by a prospective randomized trial. Wider implications of the findings Using machine-learning based analysis of patient data from IVF cycles, we were able to predict patients at risk for premature LH-rise and/or LH-surge. Interestingly, the prediction of this model was strongest in young patients. Utilizing this model will help prevent IVF cycle cancellation and better timing of ovulation triggering. Trial registration number NA