Measurement Of Luteinizing Hormone Research Articles

Correlation of serum anti-Müllerian hormone (AMH) levels on identification of premenopausal patients (pts) with hormone receptor positive (HR+), HER2-negative, node-positive breast cancer most likely to benefit from adjuvant chemotherapy in SWOG S1007 (RxPONDER).

505 Background: SWOG S1007 was a phase 3 randomized trial to evaluate the benefit of endocrine therapy alone (ET) or with chemotherapy (CET) in pts with HR+/HER2- invasive breast cancer with recurrence score (RS) ≤ 25. There was no invasive disease-free survival (IDFS) benefit for CET compared to ET in postmenopausal women However, in premenopausal women CET had an IDFS improvement compared to ET (HR=0.60; 95% 0.43-0.83). The objective was to further refine patient subgroups who benefit from CET, we measured baseline hormone levels in S1007 pts to quantify their menstrual status by serum hormone levels and associated clinical outcomes. Methods: Serum estradiol (E), progesterone (P), follicular stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH), and inhibin B (IB) were assessed on baseline serum samples from 1,015 pts who self-reported being premenopausal and age < 55. Magnetic fluorescent bead-based immunoassays ( i.e., Luminex assays) or colorimetric ELISAs were performed at the Biomarker Discovery Laboratory at the University of Kansas Medical Center to measure hormone levels. Associations of the markers (both continuous and dichotomized) with IDFS and distant relapse-free survival (DRFS) were tested for prognosis and prediction of CET benefit using Cox regression. Results: Single baseline measurement of IB, P, E, LH, FSH, or combination of E/LH/FSH, was not prognostic for IDFS or predictive for chemotherapy benefit. However, AMH levels (≥10 pg/mL) showed significant interaction with chemotherapy benefit (p=0.019), and risk increased with continuous AMH with ET alone, but not CET (p=0.03). Compared to ET alone, CET was beneficial in the 79% of premenopausal patients with AMH levels ≥10 pg/mL, a standard cutoff to define normal ovarian reserve (HR=0.48; 95% 0.33-0.69, adjusting for RS). CET was not beneficial in the 21% with AMH < 10 (HR=1.21; 95% 0.60-2.43). Absolute 5-year IDFS benefit of CET for RS ≤ 25 was 7.8% (95.2%-87.4%) for AMH ≥ 10 vs. -1.7% (89.1%-90.8%) for AMH < 10. DRFS showed a similar pattern with CET benefit in AMH ≥10 pg/mL (5-year DRFS benefit: 4.4%; HR=0.41; 95% 0.25-0.68, adjusting for RS) and no benefit in AMH < 10 (5-year DRFS benefit: -0.9%, HR=1.50; 95% 0.62-3.63). Conclusions: Among self-reported premenopausal patients < age 55, pts with baseline serum AMH levels ≥10 pg/mL showed significant benefit from CET compared to patients with AMH levels <10 pg/mL. AMH levels were a better indicator of CET benefit than self-reported menopause status, age, or E/LH/FSH levels. Use of AMH-based selection of patients may inform who most benefits from the addition of adjuvant chemotherapy to ET. Clinical trial information: NCT01272037 .

HormoneBayes: A novel Bayesian framework for the analysis of pulsatile hormone dynamics.

The hypothalamus is the central regulator of reproductive hormone secretion. Pulsatile secretion of gonadotropin releasing hormone (GnRH) is fundamental to physiological stimulation of the pituitary gland to release luteinizing hormone (LH) and follicle stimulating hormone (FSH). Furthermore, GnRH pulsatility is altered in common reproductive disorders such as polycystic ovary syndrome (PCOS) and hypothalamic amenorrhea (HA). LH is measured routinely in clinical practice using an automated chemiluminescent immunoassay method and is the gold standard surrogate marker of GnRH. LH can be measured at frequent intervals (e.g., 10 minutely) to assess GnRH/LH pulsatility. However, this is rarely done in clinical practice because it is resource intensive, and there is no open-access, graphical interface software for computational analysis of the LH data available to clinicians. Here we present hormoneBayes, a novel open-access Bayesian framework that can be easily applied to reliably analyze serial LH measurements to assess LH pulsatility. The framework utilizes parsimonious models to simulate hypothalamic signals that drive LH dynamics, together with state-of-the-art (sequential) Monte-Carlo methods to infer key parameters and latent hypothalamic dynamics. We show that this method provides estimates for key pulse parameters including inter-pulse interval, secretion and clearance rates and identifies LH pulses in line with the widely used deconvolution method. We show that these parameters can distinguish LH pulsatility in different clinical contexts including in reproductive health and disease in men and women (e.g., healthy men, healthy women before and after menopause, women with HA or PCOS). A further advantage of hormoneBayes is that our mathematical approach provides a quantified estimation of uncertainty. Our framework will complement methods enabling real-time in-vivo hormone monitoring and therefore has the potential to assist translation of personalized, data-driven, clinical care of patients presenting with conditions of reproductive hormone dysfunction.

Point-of-care impedimetric aptasensor to detect the luteinizing hormone.

Luteinizing hormone (LH) is a useful biomarker for identifying ovulation events in the cows to predict the time of ovulation to achieve a high success rate of conception following artificial insemination. Although antibody-based radioimmunoassay and enzyme-linked immunosorbent assay are being used for LH measurement, these techniques are expensive, time-consuming, and require expertise and sophisticated laboratory facilities. So, there is a need for a field-applicable, affordable, easy-to-use method for LH detection. For developing such a specific, quantitative, and inexpensive system, an aptamer-based smartphone-enabled aptasensor has been investigated. Theaptamer was used instead of the antibody as a biorecognition element due to its comparative stability at ambient temperature, ease of synthesis, and cost-effectiveness. Electrochemical impedance spectroscopy has been used to obtain label-free detection of LH within 20 min in ~ 20 μL sample volume. The screen-printed gold electrode is compatible with a smartphone-enabled miniaturized device (Sensit Smart; Palmsens BV, The Netherlands) and was fabricated with the aptamer to detect LH in biological fluids (limit of detection 0.80 and 0.61 ng/mL in buffer and undiluted/unprocessed serum, respectively, with the dynamic range of detection of 0.01 to 50 ng/mL). All the data were obtained in the 10 kHz to 0.10 Hz frequency range at a bias potential of 0.30 V with an alternating potential of 10 mV. The clinical relevance of the sensor was evaluated in 10 serum samples collected from dairy animals which established a high correlation with standard LH-ELISA (κ > 0.87). The aptasensor can be stored at room temperature for 30 days without any significant loss in electrochemical sensing ability.

Development of external genitalia during mini-puberty: is it related to somatic growth or reproductive hormones?

Although hypothalamo-pituitary-gonadal axis is active during mini-puberty, its relationship with somatic growth and the role on the development of external genitalia has not been fully elucidated. We aimed to evaluate the effects of somatic growth and reproductive hormones on the development of external genitalia during mini-puberty. Anthropometric data, pubertal assesment, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), sex-hormone binding globulin (SHBG), estradiol (E2) and inhibin-B, testosterone (T), and anti-Mullerian Hormone (AMH) of healthy infants aged 1-4months were evaluated. Free sex hormone index was calculated as T/SHBG for boys and E2/SHBG for girls. The mean age of 148 (74 female) infants included in the study was 2.31 ± 0.76months. Tanner stage 2-3 sex steroid and gonadotropin levels were observed. A statistically significant difference was found between the weight, height, BMI, weight gain and serum FSH, LH, and A4 measurements of girls and boys (p < 0.05). Penile length was associated with weight (r = 0.24, p = 0.03), height (r = 0.25, p = 0.02), and AMH (r = 0.3, p = 0.01), but not with testosterone (p = 0.56 respectively). A negative correlation was found between weight and serum LH (r = - 0.26, p = 0.2) and T/SHBG levels in males (r = - 0.38, p = 0.015 respectively). Weight-SDS was negatively correlated with testosterone in males (r = - 0.25, p = 0.02). Testicular size and breast stage did not correlate with any of the hormonal and anthropometric parameters. Conclusions: External genitalia in males during mini-puberty is related more to somatic growth rather than reproductive hormones. Similar to pubertal developmental stages, both total and free testosterone are negatively associated with higher weight during mini-puberty. What is Known: • Mini-puberty allows early assessment of HPG axis function in infancy. • There is an inverse relationship between the amount of adipose tissue and circulating testosterone levels in males during puberty and adulthood. • The potential effect of somatic growth and reproductive hormones on external genital development during mini-puberty remains unclear. What is New: • During mini-puberty, males' external genitalia is more related to somatic growth than to reproductive hormones, but this relationship is not observed in girls. • Both total and free testosterone are negatively associated with higher weight during mini-puberty, similar to the pubertal developmental stages.

Concordance of hemoglobin A1c and reproductive hormone levels in menstrual and venous blood

ObjectiveTo explore whether menstrual blood collected via a modified menstrual pad is a surrogate for venous blood draw in analyzing HbA1c and fertility-associated hormones DesignCross-sectional study SubjectsStudy included 152 female participants who have regular menses, aged 19-50 years old. InterventionsParticipants collected menstrual effluent using a menstrual pad, modified with a removable dried blood spot (DBS) strip. Peripheral blood samples were collected by venipuncture within 60 hours of menstrual pad use. Main Outcome MeasuresMenstrual pad and venous blood draw samples were analyzed for levels of HbA1c, thyroid stimulating hormone (TSH), Follicle Stimulating Hormone (FSH), anti-Mullerian hormone (AMH), and Luteinizing hormone (LH). Correlation between menstrual pad and venipuncture samples was performed by Deming linear regression, and r coefficients were measured using Pearson correlation. ResultsInter-assay variability of menstrual pad DBS sample measurements was <6%. Menstrual HbA1c values were stabilized in the DBS strips through 53 days, and menstrual hormone levels remained stable through 15 days. Menstrual HbA1c levels were highly correlated with venipuncture samples (r=0.96). TSH (r=0.94), AMH (r=0.94), FSH (r=0.91), and LH (r=0.91) also showed high correlation between menstrual strip and venipuncture samples. ConclusionsHbA1c, TSH, AMH, FSH, and LH measurements in menstrual effluent showed high correlation to venous blood samples, supporting use of menstrual effluent as a surrogate sample for hormone testing.

FRI280 Partial Loss Of KNDy Neurons In Prenatally Androgen Treated Female Rats Alters The LH Secretion And Ovarian Morphology In A Model Of Polycystic Ovary Syndrome

Abstract Disclosure: N.S. Aquino: None. A. Campideli-Santana: None. L.M. Antunes: None. R. Araújo-Lopes: None. K.S. Silva: None. P.C. Henriques: None. D.D. Gusmão: None. M.P. Bernuci: None. R.E. Szawka: None. A.M. dos Reis: None. Polycystic Ovary Syndrome (PCOS) is a high-prevalent endocrine dysfunction, causing metabolic disorders and infertility. Women with PCOS show a disrupted hypothalamus-pituitary-gonadal (HPG) axis, which results in increased luteinizing hormone (LH) secretion, LH pulse frequency, and LH/follicle stimulating hormone (FSH) ratio. These changes are related to an impaired negative-feedback effect of ovarian steroids, estradiol and progesterone, on the gonadotrophin releasing-hormone (GnRH) neurons. Because the kisspeptin/neurokinin-B/dynorphin (KNDy) neurons of the arcuate (ARC) nucleus are responsible for mediating the ovarian steroid negative feedback, we investigated whether the ablation of KNDy neurons would change the LH secretion and ovarian morphology in a rat model of PCOS. Pregnant dams were treated with dihydrotestosterone (DHT, 3 mg/day; s.c) from days 16 to 19 of pregnancy. Three months after birth, the prenatally androgen-treated (PNA) offspring received intra-ARC stereotaxic injections of the neurokinin-3 receptor agonist conjugated with saporin (NK3-SAP) to induce the lesion of KNDy neurons and were submitted to serial blood sampling for LH measurement. PNA NK3-SAP rats had a moderate loss of ARC neurokinin-B-immunoreactive neurons (between 50-70%) compared with the Blank-SAP-injected PNA group. The PNA Blank-SAP rats displayed LH pulse frequency, pulse amplitude, and mean LH levels similar to control rats on diestrus. Remarkably, the partial loss of KNDy neurons increased the LH pulse amplitude and mean LH without changing the pulse frequency. Histological analysis was performed to investigate the outcome of the partial loss of KNDy neurons on the ovarian morphology in PNA rats. The numbers of primordial, primary, and secondary health follicles were decreased in PNA Blank-SAP compared with diestrus rats. Notably, the number of primordial follicles was partially restored in NK3-SAP rats. On the other hand, neither the number of atretic follicles nor the number of corpora lutea was changed in PNA Blank or NK3-SAP rats, indicating no change in the ovulatory function. Therefore, we provide evidence that KNDy neurons negatively modulate LH release and LH pulse amplitude in PNA rats. Moreover, the results suggest that KNDy neurons are involved in the reduction of the primordial follicle pool and ovarian reserve in PNA rats. Financial Support: Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Brazilian National Council for Scientific and Technological Development (CNPq), British Society for Neuroendocrinology (BSN). Presentation: Friday, June 16, 2023

P-607 Interindividual variation of progesterone elevation post LH rise: Implications for natural cycle frozen embryo transfers in the individualized medicine era.

Abstract Study question To what extent does the period between LH-rise and P4-rise vary in ovulatory natural menstrual cycles? Summary answer Three distinct ovulatory patterns were noted defined as the period between LH-rise and P4-rise and varied by up to two days. What is known already The key to success of FET in a NC is the accurate diagnosis of ovulation to determine the optimal timing of embryo transfer. The most common method used to determine time of ovulation in NC is the detection of luteinizing hormone (LH) surge. It is well recognized that the endocrine profile of menstrual cycles varies not only amongst women but also from cycle to cycle for any given female. Most hormone trajectories in individual women differ considerably from the mean hormonal curves and LH surges culminating in ovulation appear to be highly variable in timing, amplitude and duration. Study design, size, duration Retrospective, observational study including 102 women who underwent endocrine monitoring during a frozen embryo transfer in a natural cycle between 1st January 2017 and 31st August 2021 in a tertiary IVF centre. Participants/materials, setting, methods Ultrasound scans were performed to monitor follicular growth. Serial measurements of serum LH, FSH, estradiol and P4 were commenced once a dominant follicle measuring ≥ 14mm was identified and on three consecutive days until (including) the day of ovulation. The LH surge was considered to have begun when the concentration rose by 180% above the most recent serum value. Progesterone concentrations of 1.0ng/ml and above were regarded as confirmation of ovulation to schedule FET. Main results and the role of chance Patients’ characteristics presented as median and (range): age 35 years (23-43), BMI 25.24kg/m2 (17.32-38.21), AMH 2.07 ng/ml (0.06-8.12). Twenty-one (20.6%) women had the LH-rise 2 days prior to P4 -rise, 71 (69.6%) had on the day immediately preceding and 10 (9.8%) on the same day of P4-rise. If FET was scheduled based on LH-rise, 30.4% of the patients would have their FET scheduled on a different day than FET scheduled according to P4-rise. The period between LH-rise and P4-rise would be approximately one day longer in 20.6% of the participants and one day shorter in 9.8% than the anticipated 24 hour period. There was a significant difference in body mass index between those women who had LH-rise 2 days prior to P4-rise (BMI 26.91 kg/m2 (range 23.74-30.75) and the women who had LH-rise on the same day (BMI 22.39 kg/m2 (range 20.91-24.96) (p = 0.03). The AMH levels were also found to be significantly lower in those women who had LH rise 2 days prior to P4-rise ( AMH 1.59 ng/ml (range: 0.82-2.09) as compared to those who had LH-rise on the same day (AMH 2.67ng/ml (range: 1.83—6.63) (p = 0.04). LH may not serve as the best benchmark to schedule FET. Limitations, reasons for caution Frozen embryo transfers were scheduled according to serum progesterone levels therefore our study does not provide a direct answer to the question whether FET scheduled according to serum progesterone levels provides higher live birth rates than FET scheduled based on urinary or serum LH levels in a natural cycle. Wider implications of the findings Variation in the period between LH rise and progesterone rise in ovulatory cycles has implications for the choice of marker for the start of secretory transformation in frozen embryo transfer cycles. The study participants are representative of the relevant population of women undergoing frozen embryo transfer in a natural cycle. Trial registration number Not Applicable

Validation of urinary reproductive hormone measurements using a novel smartphone connected reader

Home use tests to monitor hormone trends during the menstrual cycle have been available over-the-counter for a long time. However, these tests often depend upon manual readouts and hence may lead to false analysis. Furthermore, a lot of these tests are also not quantitative. The aim of this study was to evaluate the accuracy of the quantitative home-based fertility monitor, Inito Fertility Monitor (IFM) and to use it to identify novel hormone trends in natural menstrual cycles. There were two aspects to our analysis: (i) Evaluating the efficacy of Inito Fertility Monitor in the measurement of urinary Estrone-3-glucuronide (E3G), Pregnanediol glucuronide (PdG) and Luteinizing hormone (LH), and (ii) A retrospective study of patients' hormone profiles using IFM. To evaluate the efficacy, the recovery percentage of the three hormones from IFM was evaluated using standard spiked solutions, the accuracy of measurement was calculated and the correlation between reproducible values from IFM and ELISA was established. During the validation of IFM, novel hormone trends were also observed. In order to reinforce the observations, a second group of 52 women was recruited. Assessment of the accuracy of IFM and evaluation of the volunteer urine samples was performed in a laboratory. Home assessment of hormone analysis was carried out using IFM. For the validation study, 100 women aged 21–45 years with cycle lengths ranging from 21 to 42 days were recruited. The participants had no previously diagnosed infertility conditions and their cycle lengths did not vary for more than 3 days from the expected cycle length. Daily first morning urine samples were collected from these 100 women. For the second group, 52 women were selected meeting the same criteria set for the validation study and IFM was provided to these women for testing at home. Coefficient of variation and recovery percentage of IFM with respect to laboratory based ELISA. Percentage occurrence of novel hormone trends and AUC analysis of a novel criteria identified for confirming ovulation. We observed that with all three hormones, IFM had an accurate recovery percentage. We found that the assay has an average CV of 5.05% in PdG measurement, 4.95% in E3G measurement and 5.57% in LH measurement. Furthermore, in predicting the concentration of E3G, PdG and LH in urine samples, we show that IFM has a high correlation with ELISA. In this study, we could also reproduce hormones trends across the menstrual cycle that have been observed by previous studies. We also identified a novel criterion for earlier confirmation of ovulation which could accurately distinguish ovulatory from anovulatory cycles with 100% specificity and had an area under the ROC curve of 0.98. In addition, we identified a new hormone trend which could be observed in 94.5% of the ovulatory cycles. The Inito Fertility Monitor is an effective tool for calculating the urinary concentrations of E3G, PdG and LH and can also be used to provide accurate fertility scores and confirm ovulation. We show that certain hormone trends associated with urinary E3G, PdG and LH could be accurately captured using IFM. In addition, we report a novel criterion for earlier confirmation of ovulation compared to existing criteria. Finally, we present a novel hormone pattern associated with most of the menstrual cycles by examining hormone profiles from the volunteers recruited for the clinical trial.Trial registration: The trial is registered at the current controlled trials ISRCTN registry #ISRCTN15534557.

LBMON233 Hormonebayes: A Novel Bayesian Toolbox For Analysis Of Pulsatile Hormone Dynamics

Abstract The hypothalamus is the central regulator of reproductive hormone secretion. Pulsatile secretion of gonadotropin releasing hormone (GnRH) is fundamental to physiological stimulation of the pituitary gland to release luteinizing hormone (LH) and follicle stimulating hormone (FSH). Furthermore, GnRH pulsatility is altered in common reproductive disorders such as polycystic ovary syndrome (PCOS) and hypothalamic amenorrhea (HA). LH is measured routinely in clinical practice using an automated chemiluminescent immunoassay method and is the gold standard surrogate marker of GnRH. LH can be measured at frequent intervals (e. g., 10 minutely) to assess GnRH/LH pulsatility. However, this is rarely done in clinical practice because it is resource intensive, and there is no user-friendly and accessible method for computational analysis of the LH data available to clinicians. Here we present hormoneBayes, a novel open-access Bayesian framework that can be easily applied to reliably analyze serial LH measurements to assess LH pulsatility. The framework utilizes parsimonious models to simulate hypothalamic signals that drive LH dynamics, together with state-of-the-art (sequential) Monte-Carlo methods to infer key parameters and latent hypothalamic dynamics. We show that this method provides estimates for key pulse parameters including inter-pulse interval, secretion and clearance rates and identifies LH pulses in line with the current gold-standard deconvolution method. We show that these parameters can distinguish LH pulsatility in different clinical contexts including in reproductive health and disease in men and women (e. g., healthy men, healthy women before and after menopause, women with HA or PCOS). A further advantage of hormoneBayes is that our mathematical approach provides a quantified estimation of uncertainty. Our framework will complement methods enabling real-time in-vivo hormone monitoring and therefore has the potential to assist translation of personalized, data-driven, clinical care of patients presenting with conditions of reproductive hormone dysfunction. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

A Modified Ultra-Sensitive ELISA for Measurement of LH in Mice.

A major obstacle to monitoring pulsatile luteinizing hormone (LH) secretion in mice has been an assay with sufficient sensitivity in small blood volumes. In 2013, Steyn and colleagues published a highly sensitive enzyme-linked immunosorbent assay (ELISA) that overcame this barrier by coupling a duo of LH antibodies effective in accurately measuring LH in 4-µL whole-blood aliquots. To address the unavailability of the original detection antibody, AFP240580Rb, we validated a replacement detection antibody, biotinylated-5303 SPRN-5, to be used within the established ELISA. This modified LH ELISA demonstrated a minimum detection limit of 0.0028 ng/mL and a limit of quantification of 0.0333 ng/mL or 0.0666 ng/mL in diluted whole-blood samples of volume 6.4 µL (1:10) or 3.2 µL (1:20), respectively. Detection antibody 5303 SPRN-5 demonstrated parallelism, high precision, and accuracy across the standard curve. LH concentrations in comparison assays, using either 5303 SPRN-5 or AFP240580Rb, were highly correlated (R2 = 0.9829) and demonstrated LH pulse profiles from gonadectomized mice that were nearly superimposable. Pulsatile LH secretion was demonstrated in gonad-intact males and diestrous females and basal LH levels measured with 5303 SPRN-5 were approximately 5-fold higher than the limit of quantification. In addition, we document utility of this new LH ELISA to accurately measure LH in whole blood or serum across multiple sampling sites, as well as in pituitary extracts, LβT2 cells, or media. In summary, the modified LH ELISA described here is highly effective in measuring LH across a range of sample types and small volumes in mice.

P-406 Ongoing pregnancy rates (OPRs) after warmed blastocyst transfer (WBT) in a true-natural cycle (t-NC) are similar using six different luteinizing hormone (LH) surge criteria

Abstract Study question Does timing of WBT in t-NC differ according to six different commonly definitions for LH surge, and if so, do differences in timing impact OPRs? Summary answer Performing WBT on follicular collapse+5 days is equivalent to LH surge +7/+8 /+9 days in terms of OPRs, using six different definitions of LH surge. What is known already Pinpointing the day of ovulation, usually by documentation of the LH surge, and less commonly by transvaginal-ultrasonography is crucial for timing WBT in t-NC to maximize reproductive success. However, there is no consensus on the definition of the LH surge, and the most commonly used six LH-surge definitions are LH ≥ 10, ≥15, ≥17, ≥20 IU/L, ≥1.8-fold, and ≥2-fold increase from baseline. The usual practice is to schedule warmed blastocyst transfer on LH-surge +6 days. Study design, size, duration Prospective monitoring of 115 WBT cycles performed during January 2017-October 2021. The goals of the study were i)to assess how frequently and to what extent there would be a change in WBT related to the day of the LH surge, using the six different definitions of LH surge, compared to follicular collapse +5 days; ii)for each definition of the LH surge to compare OPRs of different WBT timings related to the day of LH surge. Participants/materials, setting, methods Staying locally and having regular menstrual cycles were the main criteria to perform t-NC. For t-NC, serial serum endocrine (LH, estradiol, and progesterone) and transvaginal ultrasonographic monitoring started on cycle days 8-10. Following precise documentation of follicular collapse by ultrasound, WBT was performed on follicular collapse +5 days. All included cycles were t-NC without human chorionic gonadotropin trigger or luteal phase support administration. Main results and the role of chance A total of 115 t-NC cycles were included for the first part of the study, determining the impact of different definitions of the LH-surge for the day of WBT. Our reference timing of follicular collapse +5 days would be equivalent to LH-surge +6 days in only 5.2%-41.2% of the cycles employing the six different LH-surge definitions. In contrast, the reference timing was comparable to LH surge +7 days in the majority of cycles (46.1%-70.8%) and less commonly to LH-surge +8 days (1.8%-38.3%) and +9 days (0%-10.4%). For the second part of the study, a total of 94 cycles were analyzed; 15 cycles were excluded as these cycles constituted 2nd or 3rd t-NC cycles; four cycles due to low serum progesterone (&amp;lt;7 ng/ml) on WBT-1 day and two cycles due to failure of survival after warming. For each LH-surge definition, OPRs were comparable among the different WBT timings related to the LH-surge (+6/+7/+8/+9 days). When logistic regression analysis was performed, taking LH-surge + 6 days as the reference, a change in timing was not an independent predictor of OPR for all six different definitions of the LH-surge. Limitations, reasons for caution Assignment of WBT timings related to LH-surge by our standard policy (follicular collapse +5 days), rather than by randomization, is a limitation. Other limitations include single daily measurements of serum LH and limited sample size. Wider implications of the findings Differences in warmed blastocyst timing related to the LH surge (LH surge +6/+7/+8/+9) are associated with comparable reproductive outcomes in t-NC, reflecting the flexibility of the window of implantation. Further, trials are warranted to delineate the best tool and timing of FET for warmed blastocyst transfer in t-NC. Trial registration number Not applicable

Gonadotropin-releasing hormone (GnRH) measurements in pituitary portal blood: A history.

Much about the neuroendocrine control of reproduction is inferred from changes in the episodic release of luteinizing hormone (LH), as measured in samples of peripheral blood. This, however, assumes that LH precisely mirrors gonadotropin-releasing hormone (GnRH) release from the hypothalamus. Because GnRH is not measurable in peripheral blood, characterization of the relationship between these two hormones required the simultaneous measurement of GnRH and LH in pituitary portal and peripheral blood, respectively. Here, we review the history of why and how portal blood collection was developed, the aspects of the true output of the central component of the hypothalamic-pituitary-gonadal axis that this methodology helped clarify, and conditions under which the pituitary fails to serve as an adequate bioassay for the release pattern of GnRH.

Socioeconomic status is related to pubertal development in a German cohort

INTRODUCTION Current health literature suggests that there has been a decline in the age of pubertal onset and that pubertal onset/duration of puberty may, besides weight status, be influenced by socioeconomic context. OBJECTIVE The goal of this study was to determine whether pubertal onset/duration and puberty-triggering hormones luteinizing hormone (LH) and follicle-stimulating hormone (FSH) vary according to socioeconomic status (SES). Moreover, we aimed to propose cutoff values of serum LH and FSH for predicting gonadarche in boys. METHODS 2,657 apparently healthy children and adolescents between 5.5 and 18 years from the area of Leipzig were recruited from the LIFE Child study. Age at pubertal onset/end of puberty was given in 738/573 children, respectively. Anthropometric parameters of puberty, blood measurements of LH and FSH, and questionnaires assessing SES were evaluated. RESULTS Lower SES was associated with earlier thelarche and longer duration of puberty in overweight/obese girls, whereas age of menarche was not affected. In boys with low SES, a trend versus earlier puberty onset can be seen. Lower SES was significantly associated with boys' age at mutation. No significant differences in boys' and girls' serum levels of LH and FSH during puberty according to SES were observed. Serum LH levels of 0.56 IU/L and serum FSH levels of 1.74 IU/L showed the best prediction of gonadarche in boys. CONCLUSION Puberty onset/duration and boys' age at mutation is affected by SES. The proposed cutoff levels for serum LH and FSH could provide a serological tool to determine gonadarche in boys.

Comparative characteristics of endocrine profile in males with infertility as well as in adolescents with varicocele

Introduction. Male infertility poses a pressing issue due to escalating prevalence of males with reproductive system diseases, the most common among which is varicocele being diagnosed in childhood and adolescence. Measuring level of hormones such as follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and estradiol is the key element while assessing the reproductive male potential.Aim: to compare the parameters of hormonal status in adolescents with varicocele as well as males with diagnosed infertility and identify predictors of hypogonadism in pubertal period.Materials and Methods. Males with diagnosed infertility as well fertile males underwent a single measurement of serum FSH, LH, testosterone and estradiol levels. Adolescents with/without varicocele were measured the serum hormones noted above in dynamics annually between studies for the period within the 14 to 17 years of age. The data obtained were compared in infertile and fertile males, adolescents with/without varicocele, as well as assessed hormone status between different age groups in both cohorts and adolescents aged 17 years.Results. Our study allowed to identify significant differences in hormone level in male patients with infertility and varicocele lacking reproductive pathology. Infertile males had significantly higher levels of LH and estradiol (for both: p &lt; 0.001). The LH level in infertile vs. fertile males was 5.01 ± 2.69 IU/L vs. 3.40 ± 1.17 IU/L, respectively, whereas estradiol level was 136.51 ± 92.79 pmol/l and 82.49 ± 48.33 pmol/l, respectively that might indicate at lowered testosterone level. Fifteenand sixteen-year-old-adolescents with varicocele had significantly lower LH level: 15 years old – 3.72 ± 1.92 IU/L vs. 2.71 ± 1.65 IU/L (p &lt; 0.0123) in subjects without vs. with varicocele; 16 years old – 3.42 ± 1.16 IU/L vs. 2.81 ± 1.66 IU/L (p &lt; 0.0381) in subjects without vs. with varicocele. It may account for decreased testosterone level. Active puberty in adolescents is accompanied by dynamically increased testosterone levels. Starting from the age of 15 years, adolescents with varicocele had significantly increased testosterone level in each subsequent age group. Thus, at 14 and 15 years of age, testosterone level in adolescents with varicocele was 10.61 ± 4.70 nmol/l and 13.60 ± 5.64 nmol/l (p &lt; 0.0001), respectively, whereas at 16 and 17 years of age it continued to rise reaching 16.65 ± 6.44 nmol/l (p &lt; 0.001) and 19.22 ± 7.36 nmol/l (p &lt; 0.0160), respectively. In contrast, age-matched adolescents without varicocele had significantly elevated testosterone level solely at age of 15 vs. 14 years. Whereas at 14 years of age testosterone level in comparison group was 15.73 ± 7.2 nmol/l, at 15 years of age it was significantly increased up to 21.45 ± 9.51 nmol/l (p &lt; 0.0113). In the subsequent age categories of this group, no significant difference in testosterone level was found. While comparing such parameter between the main and control adolescent groups, testosterone level was significantly higher in adolescents lacking varicocele at the age of 14, 15 and 16 years compared with age-matched subjects with varicocele. At 14 years of age, testosterone level in adolescents without/with varicocele was 15.73 ± 7.2 nmol/l vs. 10.61 ± 4.7 nmol/l (p &lt; 0.0007), respectively, at 15 years of age – 21.45 ± 9.57 nmol/l vs. 13.60 ± 5.64 nmol/l (p &lt; 0.0001), respectively, at 16 years of age – 20.02 ± 5.84 nmol/l vs. 16.65 ± 6.44 nmol/l (p &lt; 0.0268), respectively. The FSH level in infertile males and adolescents with varicocele was 5.16 ± 2.67 IU/L and 4.1 ±2.63 IU/L (p &lt; 0.0081), whereas LH level was 5.01 ± 2.69 IU/L and 2.76 ± 1.65 IU/L (p &lt; 0.04), respectively. In adolescents with varicocele and infertile males, estradiol level was 177.45 ± 70.63 pmol/l and 136.51 ± 92.79 pmol/l (p &lt; 0.001), respectively. While comparing hormone levels in fertile males and adolescents lacking varicocele, a significantly higher estradiol level was found in adolescents 181,87 ± 27.14 pmol/l vs. 82.49 ± 48.33 pmol/l (p &lt; 0.001).Conclusion. A study of the hormonal status in infertile vs. fertile males revealed decreased testosterone production accompanied with higher levels of LH and estradiol. Due to profound changes in LH and testosterone levels detected in adolescents with varicocele as well as in infertile males, it is plausible that such hormones may serve as predictors of hypogonadism in the pubertal period. Adolescents with varicocele taking into consideration progressive course of the disease and verified lower testosterone level require further monitoring of hormone level to prevent endocrine infertility.

Socioeconomic Status Is Related to Pubertal Development in a German Cohort

Introduction: Current health literature suggests that there has been a decline in the age of pubertal onset and that pubertal onset/duration of puberty may, besides weight status, be influenced by socioeconomic context. Objective: The goal of this study was to determine whether pubertal onset/duration and puberty-triggering hormones luteinizing hormone (LH) and follicle-stimulating hormone (FSH) vary according to socioeconomic status (SES). Moreover, we aimed to propose cutoff values of serum LH and FSH for predicting gonadarche in boys. Methods: 2,657 apparently healthy children and adolescents between 5.5 and 18 years from the area of Leipzig were recruited from the LIFE Child study. Age at pubertal onset/end of puberty was given in 738/573 children, respectively. Anthropometric parameters of puberty, blood measurements of LH and FSH, and questionnaires assessing SES were evaluated. Results: Lower SES was associated with earlier thelarche and longer duration of puberty in overweight/obese girls, whereas age of menarche was not affected. In boys with low SES, a trend versus earlier puberty onset can be seen. Lower SES was significantly associated with boys’ age at mutation. No significant differences in boys’ and girls’ serum levels of LH and FSH during puberty according to SES were observed. Serum LH levels of 0.56 IU/L and serum FSH levels of 1.74 IU/L showed the best prediction of gonadarche in boys. Conclusion: Puberty onset/duration and boys’ age at mutation is affected by SES. The proposed cutoff levels for serum LH and FSH could provide a serological tool to determine gonadarche in boys.

Does the repeat dose of gonadotropin-releasing hormone agonist trigger in polycystic ovarian syndrome improve in vitro fertilization cycles outcome? A clinical trial study.

Background A repeat dose of Gonadotropin-releasing Hormone (GnRH) agonist could provide long duration of luteinizing hormone (LH) surge and amplitude appropriately.Objective Improvement in oocyte maturity could be obtained by a repeat dose of GnRH agonist.Materials and MethodsIn this randomized double-blinded study, 120 women with polycystic ovarian syndrome and serum estradiol level (E2) 3000 who were candidate for in vitro fertilization with Antagonist protocol were enrolled between July 2018 and July 2019. Participants were randomized in two groups - and final oocyte maturation was triggered with two doses: In group A, a repeat dose of 0.1 mg, 12 hr. after the first dose and in group B, 0.2 mg SC triptorelin (decapeptyl) 35 hr. prior to oocyte retrieval. Serum Estradiol, LH, and progesterone concentration were measured on the trigger day. Serum LH measurement was done three times in both groups. The outcomes were oocyte yield, meiosis (M) I, MII, Maturity rate, germinal vesicle (GV) rate, 2 pronuclear, embryo yield, ovarian hyper stimulation syndrome rates.ResultsMaturity rate (p = 0.89), MI (p = 0.38), MII (p = 0.89), and GV oocytes (p = 0.38) were not statistically different between the two study groups. LH levels measured at 12 hr post-trigger did not relate statistically significant with maturity rate in our participants (p = 0.96). No empty follicular syndrome was reported.ConclusionAlthough, the second dose of GnRH agonist after 12 hr since the first dose could provide duration of LH surge and amplitude and as a result no empty follicular syndrome was seen, the maturity rate, MI, MII, and GV oocytes were not different between the two study groups.

Estradiol Potentiates But Is Not Essential for Prolactin-Induced Suppression of Luteinizing Hormone Pulses in Female Rats.

Hyperprolactinemia causes infertility by suppressing gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion. Because effects of prolactin (PRL) on the hypothalamus usually require estradiol (E2), we investigated the role of E2 in PRL-induced suppression of LH pulses. Ovariectomized (OVX) rats treated with oil or E2 (OVX + E2) received a subcutaneous injection of ovine PRL (oPRL) 30 minutes before serial measurement of LH in the tail blood by enzyme-linked immunosorbent assay. E2 reduced pulsatile LH secretion. oPRL at 1.5 mg/kg further reduced LH pulse frequency in OVX + E2 but had no effect in OVX rats. The higher dose of 6-mg/kg oPRL decreased LH pulse frequency in both OVX and OVX + E2 rats, whereas pulse amplitude and mean LH levels were lowered only in OVX + E2 rats. Kisspeptin immunoreactivity and Kiss1 messenger ribonucleic acid (mRNA) levels were decreased in the arcuate nucleus (ARC) of OVX + E2 rats. oPRL decreased both kisspeptin peptide and gene expression in the ARC of OVX rats but did not alter the already low levels in OVX + E2 rats. In the anteroventral periventricular nucleus, oPRL did not change kisspeptin immunoreactivity and, paradoxically, increased Kiss1 mRNA only in OVX + E2 rats. Moreover, oPRL effectively reduced Gnrh expression regardless of E2 treatment. In this study we used tail-tip blood sampling to determine the acute effect of PRL on LH pulsatility in female rats. Our findings characterize the role of E2 in the PRL modulation of hypothalamic components of the gonadal axis and LH release, demonstrating that E2 potentiates but is not essential for the suppression of pulsatile LH secretion caused by hyperprolactinemia.

Analysis of luteinizing hormone (LH): Validation of a commercial ELISA kit for LH analysis and quantification in doping control samples.

Luteinizing hormone (LH) is a dimeric glycoprotein produced and secreted by the pituitary gland, with a molecular weight of approximately 30 000 Da. The main clinical use for exogenous LH-administration is typically linked to the treatment of infertility, in both men and women. The desired effect of LH misuse in sport is due to the enhancement of testicular production of testosterone. Elevated LH levels may also indicate the usage of gonadotropin-releasing factors or estrogen blockers. Therefore, LH is listed by the World Anti-Doping Agency (WADA) as a prohibited substance in male athletes, and according to the WADA technical document, laboratories should determine the the total LH concentrations in urine. The TD lists two different assays that are suitable for measuring total LH in urine, Delfia and Siemens Immulite. Other fit-for-purpose assays can be used, as long as they are capable of detecting total LH in urine. In case an assay not listed in the TD will be used, population-based reference values have to be determined in the validation procedure. In this study a new immunoassay was validated for the measurement of LH in urine. The assay (AccuBind ELISA Microwells, Luteinizing Hormone, Monobind Inc.), originally intended for serum, showed adequate sensitivity and was proven fit-for-purpose in routine doping control. Population-based distribution of the assay was in good agreement with the results of Delfia and Immulite assays, for which the method-specific cut-off-values are 40 IU/L (Delfia) and 60 IU/L (Siemens Immulite).

Activation of a Classic Hunger Circuit Slows Luteinizing Hormone Pulsatility

Introduction: The central regulation of fertility is carefully coordinated with energy homeostasis, and infertility is frequently the outcome of energy imbalance. Neurons in the hypothalamus expressing neuropeptide Y and agouti-related peptide (NPY/AgRP neurons) are strongly implicated in linking metabolic cues with fertility regulation. Objective: We aimed here to determine the impact of selectively activating NPY/AgRP neurons, critical regulators of metabolism, on the activity of luteinizing hormone (LH) pulse generation. Methods: We employed a suite of in vivo optogenetic and chemogenetic approaches with serial measurements of LH to determine the impact of selectively activating NPY/AgRP neurons on dynamic LH secretion. In addition, electrophysiological studies in ex vivo brain slices were employed to ascertain the functional impact of activating NPY/AgRP neurons on gonadotropin-releasing hormone (GnRH) neurons. Results: Selective activation of NPY/AgRP neurons significantly decreased post-castration LH secretion. This was observed in males and females, as well as in prenatally androgenized females that recapitulate the persistently elevated LH pulse frequency characteristic of polycystic ovary syndrome (PCOS). Reduced LH pulse frequency was also observed when optogenetic stimulation was restricted to NPY/AgRP fiber projections surrounding GnRH neuron cell bodies in the rostral preoptic area. However, electrophysiological studies in ex vivo brain slices indicated these effects were likely to be indirect. Conclusions: These data demonstrate the ability of NPY/AgRP neuronal signaling to modulate and, specifically, reduce GnRH/LH pulse generation. The findings suggest a mechanism by which increased activity of this hunger circuit, in response to negative energy balance, mediates impaired fertility in otherwise reproductively fit states, and highlight a potential mechanism to slow LH pulsatility in female infertility disorders, such as PCOS, that are associated with hyperactive LH secretion.

Elevated Random Luteinizing Hormone is an Unreliable Indicator for Pubertal Suppression in Girls Treated with Monthly Leuprolide for Idiopathic Central Precocious Puberty

Objective:Longitudinal data regarding random luteinizing hormone (LH) concentrations in patients with idiopathic central precocious puberty (ICPP) during treatment are limited. Therefore, we sought to evaluate random LH and estradiol concentrations during monthly leuprolide injection and their associations with pubertal progression and final adult height (FAH) in girls with ICPP.Methods:Medical records of 27 girls with ICPP who had attained FAH were reviewed. Patients’ height, weight, Tanner stage, growth rate (GR), bone age, random LH measured by both immunoradiometric and immunochemiluminescent methods, follicular-stimulating hormone (FSH) and estradiol levels were monitored until FAH.Results:Treatment was started at a mean (±standard deviation) age of 8.1±0.6 years with mean duration of 3.9±0.2 years. At six months of follow-up, random LH (p=0.048), FSH (p<0.001) and estradiol (p=0.023) concentrations were decreased compared with baseline. Thereafter, random LHs were well suppressed. GRs gradually decreased to prepubertal norm by month 12. Seventeen patients (63%) exhibited pubertal LH concentrations at least once during treatment visits. Furthermore, 43 of a total 116 (37%) LH measurements were found elevated. However, those patients with elevated random LH did not show signs of pubertal progression. After treatment, mean FAH was greater than predicted adult height (p<0.0001) and target height (p=0.03). At no time points of treatment did random LH, FSH and estradiol correlate with GRs or FAH.Conclusion:Elevated random LH is commonly found in ICPP girls during monthly leuprolide treatment. However, these elevations were not associated with clinical progression of puberty or decreased FAH, suggesting that it is not a reliable method for CPP monitoring.