Mean Luteinizing Hormone Research Articles

INCIDENCE, SOCIODEMOGRAPHIC, AND CLINICAL CHARACTERISTICS OF ANOVULATORY INFERTILITY AMONG WOMEN PRESENTING TO A SECONDARY CARE HOSPITAL OF NORTH INDIA

Objectives: The objective of the study was to assess the incidence and factors associated with anovulatory infertility among reproductive-age women. Methods: The study was an observational cross-sectional study conducted in the Department of Obstetrics and Gynecology of a secondary care hospital of Prayagraj, Uttar Pradesh over a period of 20 months, from October 2020 to May 2022. A consecutive sampling method was used to recruit a total of 100 women presenting with a complaint of inability to conceive even after regular intercourse over the past 1 year into the present research. Based on the detailed physical and clinical examination as well as laboratory findings, the final diagnosis of the cause and the nature of the infertility (primary/secondary) were established. Results: Among the 100 patients, 22 were found to have anovulatory infertility. The mean age was 28.68±3.71 years. Majority (50%) belonged to the upper socioeconomic class the mean weight of patients was 55.91±5.66 kg and the mean body mass index was 22.85±2.21 kg/m2. Majority presented with normal menstruation, followed by oligomenorrhoea (45.5%). 81.8% of the participants had hirsutism. The mean follicle-stimulating hormone of patients was 6.30±1.87 IU/mL, the mean luteinizing hormone was 6.04±1.83 IU/mL, mean estradiol was 45.00±6.50 pg/mL, mean anti-mullerian hormone was 6.94±2.13 ng/mL, mean total cholesterol was 76.21±16.86 ng/dL, mean thyroid stimulating hormone was 2.63±0.98 mIU/L and mean prolactin was 15.94±4.94 ng/mL. About 86.4% had primary infertility, the most common cause being polycystic ovarian syndrome (PCOS, 63.6%). This was followed by hyperprolactinemia (13.6%) and hypothyroidism (9.2%). Secondary anovulation associated with obesity was identified in 13.6% of the patients. Conclusion: The incidence of anovulatory infertility was 22%, with the majority being primary infertility due to PCOS, followed by hyperprolactinemia, and hypothyroidism.

Central δ/κ opioid receptor signaling pathways mediate chronic and/or acute suckling-induced LH suppression in rats during late lactation.

In mammals, secretion of tonic (pulsatile) gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) is often suppressed during lactation. Suppression of GnRH/LH pulses in lactating dams is assumed to be caused by suckling stimuli and a chronic negative energy balance due to milk production. The present study aimed to investigate whether the central enkephalin-δ opioid receptor (DOR) signaling mediated the suppression of LH secretion by acute suckling stimuli and/or chronic negative energy balance due to milk production in rats during late lactation when dams were under a heavy energy demand. On postpartum day 16, the number of Penk (enkephalin mRNA)-expressing cells in the arcuate nucleus was significantly higher in lactating rats than in non-lactating control rats. Pulsatile LH secretion was suppressed in rats with chronic suckling or acute 1-h suckling stimuli 6 h after pup removal on day 16 of lactation. Central DOR antagonism significantly increased the mean LH concentrations and the baseline of LH pulses in rats with chronic suckling but not with acute suckling stimuli on day 16 of lactation. Besides, central κ opioid receptor (KOR) antagonism increased the amplitude of LH pulses in rats with the acute suckling stimuli on day 16 of lactation. These results suggest that central DOR signaling mediates the suppression of LH secretion caused by a negative energy balance in rats receiving chronic suckling during late lactation. On the other hand, central KOR signaling likely mediates acute suckling stimuli-induced suppression of LH secretion in rats during late lactation.

Chemogenetic Activation of RFRP Neurons Reduces LH Pulse Frequency in Female but not Male Mice.

The neuropeptide RFRP-3 (RFamide-related peptide-3) is thought to play a role in the negative regulation of fertility. However, the exogenous administration of RFRP-3 yields varying results depending on the dose and route of administration, sex of the subject, and many other variables. Manipulation of in vivo neuronal activity using DREADDs (designer receptor exclusively activated by designer drugs) technology enables investigation of cell type-specific neuronal activation in a manner that better reflects endogenous neuronal activity. To test the effects of RFRP neuronal activation on pulsatile luteinizing hormone (LH) secretion. We generated mice expressing the stimulatory hM3Dq designer receptor exclusively in RFRP cells using 2 different Cre-loxP-mediated approaches: (1) we bred mice to express hM3Dq in all Rfrp-Cre-expressing cells, including some that transiently expressed Rfrp-Cre neonatally (RFRP × hM3Dq mice), and (2) we stereotaxically injected Cre-dependent hM3Dq into the dorsomedial nucleus of RFRP-Cre mice to drive hM3Dq expression exclusively in a subpopulation of adult Rfrp-Cre neurons (RFRP-AAV-hM3Dq mice). We then investigated the effects of acute hM3Dq activation on LH pulse frequency in RFRP × hM3Dq mice, RFRP-AAV-hM3Dq mice, and their respective controls. In both female RFRP × hM3Dq and RFRP-AAV-hM3Dq mice, chemogenetic activation of Cre-driven hM3Dq led to a significant 35% to 50% reduction in LH pulse frequency compared with controls, while no differences in pulse amplitude or mean LH concentration were observed. In marked contrast, RFRP activation did not cause any changes to LH pulse dynamics in male mice. These data show for the first time that activation of neurons that have expressed Rfrp, or of a subset of adult RFRP neurons, can independently suppress LH pulsatility in female, but not male mice.

Peripheral administration of a κ-opioid receptor agonist nalfurafine inactivates gonadotropin-releasing hormone pulse generator activity in goats

Neurons co-expressing kisspeptin, neurokinin B, and dynorphin A (KNDy neurons), located in the arcuate nucleus (ARC) of the hypothalamus, are indicated to be the gonadotropin-releasing hormone (GnRH) pulse generator. Dynorphin A is reported to suppress GnRH pulse generator activity. Nalfurafine is a selective agonist of the κ-opioid receptor (KOR), a receptor for dynorphin A, clinically used as an anti-pruritic drug. This study aimed to evaluate the effects of nalfurafine on GnRH pulse generator activity and luteinizing hormone (LH) pulses using female goats. Nalfurafine (0, 2, 4, 8, or 16 μg/head) was intravenously injected into ovariectomized Shiba goats. The multiple unit activity (MUA) in the ARC area was recorded, and plasma LH concentrations were measured 2 and 48 h before and after injection, respectively. The MUA volley interval during 0–2 h after injection was significantly increased in the nalfurafine 8 and 16 μg groups compared with the vehicle group. In 0–2 h after injection, the number of LH pulses was significantly decreased in the nalfurafine 8 and 16 μg groups, and the mean and baseline LH were significantly decreased in all nalfurafine-treated groups (2, 4, 8, and 16 μg) compared with the vehicle group. These results suggest that nalfurafine inhibits the activity of the GnRH pulse generator in the ARC, thus suppressing pulsatile LH secretion. Therefore, nalfurafine could be used as a reproductive inhibitor in mammals.

Serum anti-mullerian hormone, sex hormone, and nutrient levels in reproductive age women with celiac disease.

This study aimed to investigate the changes in serum Anti-Müllerian Hormone (AMH) levels, sex hormone levels, follicle-stimulating hormone (FSH)/luteinizing hormone (LH) ratio in patients with celiac disease (CeD), and their correlation with clinical characteristics and nutrient levels. This cross-sectional study collected clinical and biochemical data from a total of 67 females diagnosed with CeD and 67 healthy females within the reproductive age range of 18-44 years. The study was conducted at a tertiary hospital between September 2016 and January 2024. Both groups underwent comprehensive clinical and laboratory assessments. Serum levels of AMH and sex hormones were quantified using chemiluminescence immunoassay, and their associations with CeD clinical features and nutrient levels were thoroughly analyzed. The study included 67 patients and 67 controls with a mean age of 36.7±7.6 years. No statistically significant differences were found between the two groups in mean age, BMI, FSH, LH, E2, P levels, FSH/LH, menstrual irregularities, abortions history, parity, and gravidity (all P>0.05). However, AMH, T, FER, FA, Zn, and Se levels were significantly lower, and PRL levels were higher in the CeD group (all P<0.05). Spearman's correlation analysis showed that AMH levels were negatively correlated with age, tTG level, disease duration, and Marsh grading (P<0.05). This study highlights the association between impaired ovarian function in CeD patients and disease severity and nutrient levels. Early detection and intervention for ovarian function abnormalities are imperative to enhance fertility potential in CeD patients.

ASSOCIATION OF POLYCYSTIC OVARY SYNDROME AND THYROID DYSFUNCTION AMONG REPRODUCTIVE WOMEN: A CROSS-SECTIONAL STUDY

Objective: Polycystic ovarian syndrome (PCOS) is became most common problem in the reproductive age group women who seem to be adversely affected by associated thyroid dysfunction. Ovarian failure and pregnancy-related complications leaded by both factors. The study helps to explore the incidence and etiology of diverse thyroid conditions among PCOS individuals. Methods: This prospective single‑center study with 40 female patients with hyperandrogenism, hirsutism, oligo‐anovulation, and polycystic ovaries patients were defined as having PCOS according to the revised 2003 Rotterdam criteria and Ferriman–Gallwey score with comprised the study population. Normal female subjects were studied as the control population who had normal thyroid function and without PCOS. By measuring of serum thyroid-stimulating hormone (TSH), free thyroxine levels (free T3 and free T4), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen, progesterone, and testosterone can evaluate thyroid function. Along with ultrasound was performed to confirm the PCOS. By SPSS version 19 Chi-square test and t-test were determined. Results: This case–control study revealed that a statistically significant higher prevalence of PCOS patients was found to have higher mean TSH values that were 5.33±2.51 μIU/mL, the mean for T3 and T4 was 154±138 μg/dL and 11.27±8.47 μg/dL, respectively. The mean FSH values were 07.27±4.49 μIU/mL, the mean LH was 84.27±38.49 μIU/mL, the mean estrogen 342±202 pg/mL, the mean progesterone values were 14.2±8.9 ng/mL and the mean testosterone levels were 69.73±9.28 ng/mL. Conclusion: Thyroid dysfunction shows a high prevalence in PCOS patients. Dysfunction can be treated which helps to manage infertility associated with PCOS.

Pubertal Development in Girls with Beta Thalassaemia and Assessment of the Adequacy of Chelation Therapy: A Quasi-experimental Study

Introduction: Beta thalassaemia is the most prevalent hereditary autosomal disorder, significantly impacting endocrine function during pubertal development. The pathology is rooted in the excessive deposition of iron in vital organs. If left untreated, this condition leads to serious morbidity and mortality. Hypogonadism stands as the most common endocrine complication. Aim: To observe pubertal development in girls with thalassaemia and evaluate the adequacy of chelation therapy in such patients by measuring levels of Follicle Stimulating Hormone (FSH), Luteinising Hormone (LH), oestrogen, and serum ferritin. Materials and Methods: A quasi-experimental study was conducted at the Thalassaemia Clinic at the Department of Obstetrics and Gynaecology, Burdwan Medical College, Purba Bardhaman, West Bengal, India involving 300 diagnosed Beta Thalassaemia Major (BTM) (case) patients aged 13 to 17 years over a period of one and a half years (January 2020 to June 2021). The girls in the study groups were sequentially enrolled from the Outpatient Department of the Thalassaemia Clinic after meeting inclusion and exclusion criteria. Demographic data, anthropometric measurements, and Sexual Maturity Rating (SMR) were recorded. Hormonal assays of serum FSH, LH, oestrogen, and serum ferritin levels were conducted. Thalassaemic girls with serum ferritin levels exceeding 500 ng/mL received chelation therapy. Serum ferritin levels were measured at two-month intervals for up to six months, with reassessment of serum FSH, LH, oestrogen, and ferritin levels at the end of the six-month period. Statistical analysis was conducted using Statistical Package for Social Sciences (SPSS) version 27.0, Microsoft excel spreadsheet, and Epi Info 7. Results: A total of 190 (63.3%) of the thalassaemic girls were aged 13 to 15 years, with a mean Body Mass Index (BMI) of 22.5000±2.6100 kg/m2 , and 74 of them (38.94%) experienced menarche in this age group. Out of 300 cases, 269 thalassaemic girls (89.6%) received chelation therapy, and 89 cases (33.09%) reached menarche after chelation therapy. A significant (p&lt;0.0001) increase in mean LH and FSH levels was observed after chelation therapy. The mean ferritin level decreased from 3168.85 ng/mL to 2227.24 ng/mL following chelation therapy. Conclusion: Pubertal development failure is common in beta thalassaemic girls. Intervention in the form of adequate chelation therapy in girls with high serum ferritin levels yielded favourable outcomes, as evident from serum gonadotropin and oestrogen levels.

A high-fat eucaloric diet induces reprometabolic syndrome of obesity in normal weight women.

We examined the effects of 1 month of a eucaloric, high-fat (48% of calories) diet (HFD) on gonadotropin secretion in normal-weight women to interrogate the role of free fatty acids and insulin in mediating the relative hypogonadotropic hypogonadism of obesity. Eighteen eumenorrheic women (body mass index [BMI] 18-25 kg/m2) were studied in the early follicular phase of the menstrual cycle before and after exposure to an HFD with frequent blood sampling for luteinizing hormone (LH) and follicle-stimulating hormone (FSH), followed by an assessment of pituitary sensitivity to gonadotropin-releasing hormone (GnRH). Mass spectrometry-based plasma metabolomic analysis was also performed. Paired testing and time-series analysis were performed as appropriate. Mean endogenous LH (unstimulated) was significantly decreased after the HFD (4.3 ± 1.0 vs. 3.8 ± 1.0, P < 0.01); mean unstimulated FSH was not changed. Both LH (10.1 ± 1.0 vs. 7.2 ± 1.0, P < 0.01) and FSH (9.5 ± 1.0 vs. 8.8 ± 1.0, P < 0.01) responses to 75 ng/kg of GnRH were reduced after the HFD. Mean LH pulse amplitude and LH interpulse interval were unaffected by the dietary exposure. Eucaloric HFD exposure did not cause weight change. Plasma metabolomics confirmed adherence with elevation of fasting free fatty acids (especially long-chain mono-, poly-, and highly unsaturated fatty acids) by the last day of the HFD. One-month exposure to an HFD successfully induced key reproductive and metabolic features of reprometabolic syndrome in normal-weight women. These data suggest that dietary factors may underlie the gonadotrope compromise seen in obesity-related subfertility and therapeutic dietary interventions, independent of weight loss, may be possible.

PSIV-17 Postnatal Obesity Amplifies Gnrh-Induced Lh Release in Prenatal Testosterone-Treated Female Sheep

Abstract Polycystic ovary syndrome (PCOS) is the most common hormonal disorder in females of reproductive age. Hypersecretion of luteinizing hormone (LH), partly due to increased pituitary sensitivity to gonadotrophin-releasing hormone (GnRH), is a significant cause of infertility in women with PCOS. Moreover, obesity amplifies the severity of reproductive dysfunction in PCOS. In sheep, testosterone (T) excess during gestational days (GD) 30 to 90 results in GnRH-mediated LH hypersecretion and recapitulates the PCOS phenotype. This study investigated 1) if mid-gestation (GD 60 to 90) is the susceptibility window for programming pituitary hypersensitivity to GnRH, and 2) the contribution of postnatal obesity in modulating LH responsiveness to GnRH. Suffolk ewes received T propionate (T; 100 mg i.m.) or corn oil (C; vehicle) twice weekly from GD 60 to 90 (term = 147 d). At 5 mo of age, T lambs were assigned randomly to either a maintenance (100% of NRC requirements) or overfed diet (130% of NRC requirements), and Control lambs were fed the maintenance diet. During the anestrous period (late summer), ewes (n = 8/group; ~17 mo of age) received a GnRH bolus injection (2 ng/kg) intravenously every 1.5 h and blood samples were collected at 15-min intervals for a 6-h period. Despite animals being in seasonal anestrous, few endogenous LH pulses were evident during the 6 h-sampling period. To avoid confounding, only clear responses to GnRH challenge that occurred within 15 to 45 min after each GnRH injection were considered for further analyses. Mean number of exogenous GnRH-induced LH responses that met these criteria did not differ between groups and averaged 1.81 ± 0.27 pulses/animal. The parameters of these GnRH-induced responses were analyzed as one-way ANOVA with Tukey’s HSD post-hoc analysis. Mean LH pulse peak was greater (P &amp;lt; 0.05) in T overfed ewes (4.59 ± 0.49 ng/mL) compared with T maintenance (2.19 ± 0.61 ng/mL) and Control ewes (2.58 ± 0.53 ng/mL). Additionally, amplitude of LH pulses was greater (P &amp;lt; 0.05) in T overfed ewes (2.66 ± 0.39 ng/mL) compared with T maintenance (1.03 ± 0.48 ng/mL) and tended (P = 0.089) to be greater in T overfed compared with Control ewes (1.41 ± 0.42 ng/mL). LH pulse peak and amplitude in T maintenance did not differ from Control ewes. In conclusion, failure of GD 60 to 90 T treatment to increase GnRH responsiveness as opposed to our earlier finding of increased responsiveness in GD 30 to 90 T-treaded sheep indicates that the susceptibility window for programming pituitary hypersensitivity to GnRH lies between GD 30 to 60. Postnatal obesity may act as an independent activational signal in amplifying the pituitary sensitivity to GnRH in female sheep. This premise is supported by observations that obesity exacerbates pituitary-mediated LH hypersecretion in women with PCOS. Research support: NIH-NICHD (R01HD099096).

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

SAT376 Luteinizing Hormone Secretion in Hyperandrogenic Girls Taking Spironolactone or Metformin

Abstract Disclosure: C.M. Burt Solorzano: None. M. Gilrain: None. S.H. Kim: None. C.R. McCartney: None. Peri-pubertal hyperandrogenism (HA) typically relates to excess adrenal and/or ovarian androgen production in response to adrenocorticotropin (ACTH) and/or luteinizing hormone (LH), respectively. Spironolactone and metformin are commonly used medications to treat adolescent androgen excess. We examined how end-organ responsiveness and LH pulse patterns are affected by these medications in adolescents with HA. We studied free testosterone (T) levels (including baseline and peaks after ACTH and recombinant human chorionic gonadotropin [r-hCG] stimulation), oral glucose tolerance testing (OGTT), and 12-hr overnight LH secretion (sampled q 10 min) in post-menarcheal girls with HA before and after 3-months of either spironolactone (100 mg BID) or metformin (500 mg BID). Free T was calculated using total T (LC-MS/MS) and sex hormone binding globulin (SHBG).To date, 3 girls (aged 12-18) have been studied using spironolactone and 4 girls (aged 14-17) using metformin. (1 girl was studied twice, once with each medication.) After spironolactone treatment, free T changed by -19% (-58 to +37) (median [range]) unstimulated, by -53% (-62 to -15) after ACTH, and by -54% (-66 to +12) after r-hCG. Peak insulin level during OGTT changed by +28% (+16 to +98). Overnight LH pulse frequency changed by -64% (-89 to -10) and LH mean by -74% (-89 to -17). Two subjects in the spironolactone group demonstrated luteal LH patterns despite serum progesterone concentrations &amp;lt; 0.7 ng/mL. In the metformin group, free T changed by +35% (-32 to +125) unstimulated, by 36% (-54 to +66) after ACTH, and by 21% (-41 to +105) after r-hCG. Peak insulin level during OGTT changed by -47% (-74 to +102). LH pulse frequency changed by +10% (-29 to +22) and LH mean by +1% (-2 to +74). To summarize findings to date, mean LH and LH pulse frequency decreased in all three subjects taking spironolactone, including marked reductions in two subjects who demonstrated luteal LH patterns. While free T levels decreased in some of these girls, this seems unlikely to account for such marked changes in LH secretion. And despite similar decreases in T and also decreases in peak insulin in some girls taking metformin, LH pulse frequency did not change markedly. LH pulse patterns in two girls taking spironolactone suggest a progesterone-like effect on LH pulsatility. Spironolactone can cause irregular uterine bleeding, and canrenone (the major metabolite of spironolactone) binds to uterine progesterone receptors competitively (Fernandez MD, Br J Clin Pharm 1983). However, to our knowledge, this is the first report of LH pulse changes in HA girls using spironolactone. Further studies are needed to confirm this finding. Presentation Date: Saturday, June 17, 2023

OR25-04 Factors Affecting Testosterone Concentrations In Men: Individual Participant Data Meta-analyses From The Androgens In Men Study

Abstract Disclosure: R.J. Marriott: None. K. Murray: None. L. Antonio: None. S. Bhasin: None. A.S. Dobs: None. D.J. Handelsman: None. G.J. Hankey: None. R. Haring: None. A.M. Matsumoto: None. C. Ohlsson: None. E.S. Orwoll: None. D.M. Vanderschueren: None. G.A. Wittert: None. F.C. Wu: None. B.B. Yeap: None. Different factors modulate circulating testosterone in men, impacting interpretation of reference ranges. We aimed to clarify factors associated with variations in sex hormone concentrations by conducting individual participant data (IPD) meta-analyses of prospective cohort studies of adult men with total testosterone measured using mass spectrometry. The Androgens In Men Study protocol was submitted to PROSPERO (23 July 2019), registered (20 November 2019; CRD42019139668) and published (BMJ Open 2020; 10: e034777). A systematic review (14 June-31 December 2019) identified prospective cohort studies (BMJ Open 2021; 11: e048013). IPD data were requested. Cross-sectional analyses related total testosterone (measured using mass spectrometry), sex hormone binding globulin (SHBG), luteinising hormone (LH), dihydrotestosterone (DHT) and estradiol (also measured using mass spectrometry) concentrations to sociodemographic, lifestyle, and health factors. Summary curves and effect estimates with 95% confidence intervals (CIs) were obtained using two-stage random-effects IPD meta-analyses. Summary estimates were obtained from 11 studies (25,149 men). There was a non-linear association of testosterone with age, with negligible change among men aged 17-70 years (1SD increase about age 43.5, from 35.7 to 51.3 years -0.27 nmol/L, CI -0.71 to 0.18) and decreasing testosterone with age for men &amp;gt;70 years (1SD increase about age 84.5, from 76.7 to 92.3 years -1.55 nmol/L, CI -2.05 to -1.06). Testosterone was inversely associated with BMI (1SD increase about 27.5 kg/m2, from 25.5 to 29.6 kg/m2 -2.42 nmol/L, CI -2.70 to -2.13). Testosterone concentrations were lower for men who: were married/de facto (-0.57 nmol/L); undertook ≤75 minutes vigorous physical activity/week (-0.51 nmol/L); former smokers (-0.34 nmol/L); had hypertension (-0.53 nmol/L), cardiovascular disease (-0.35 nmol/L), cancer (-1.39 nmol/L), or diabetes (-1.43 nmol/L); all CIs excluded zero. SHBG increased with age (per SD increase in age 11.3 nmol/L, CI 9.0 to 13.6) and decreased with BMI (per SD increase in BMI -5.9 nmol/L, CI -6.9 to -5.0). There was little change in mean LH with age in men &amp;lt;70 years (1SD increase in age 0.10 IU/L, CI -0.08 to 0.28), but an increase in LH with age in men ≥70 years (1SD increase in age 4.14 IU/L, CI 3.71 to 4.56). DHT and estradiol were less prominently associated with these factors. Multiple factors are associated with variation in male testosterone and SHBG concentrations, with evidence of primary impairment of testicular hormone production after age 70 years. Interpretation of testosterone results in individuals needs to account for these factors, especially age &amp;gt;70 years, BMI, diabetes and cancer. Further research is needed to determine health impacts of declining testosterone in older men. Presentation Date: Saturday, June 17, 2023

SAT606 Comparison Of Psychosocial And Immune/Inflammatory Stress On Luteinizing Hormone Pulsatile Secretion

Abstract Disclosure: J. Jang: None. M.J. Kreisman: None. K.M. Breen Church: None. Stress induces a physiological response in the body to return to the homeostatic state. It is recognized that many stress types impair reproduction, but the mechanisms conveying inhibition may be distinct and stressor-dependent. Our research aims to understand how stress inhibits the neuroendocrine mechanisms controlling the pulsatile release of luteinizing hormone (LH). This is especially important because pulsatile LH is pivotal to reproductive health in males and females. Immune/inflammatory stressors such as lipopolysaccharides (LPS) cause an inflammatory response, producing peripheral cytokines that can signal to the brain in distinct ways compared to the neural mechanisms employed in response to psychosocial stress. However, how LPS disrupts the neural mechanisms involved in gonadotropin pulsatility is still unknown. Here, we compared the alteration in LH pulses in response to psychosocial restraint stress or immune/inflammatory LPS stress and sought to understand stress-specific regulation of the upstream targets controlling LH synthesis and secretion. Our initial observations demonstrated, in ovariectomized (OVX) female mice, that restraint stress elicited suppression rapidly, within 30 minutes; yet, the response to LPS was delayed, occurring approximately 90 minutes from injection. Therefore, we investigated the prolonged response of the stressors on LH pulses, observing LH pulse profiles 90 minutes to 180 minutes after the initiation of stress. Restraint stress significantly suppressed mean LH, resulting from a suppression of pulse frequency. LPS injected intraperitoneal (IP, 1.5 ug/g) also significantly suppressed mean LH. In contrast to restraint stress, LPS induced a reduction in both frequency as well as LH pulse amplitude. We conducted two follow-up experiments to investigate the lowered LH pulse amplitude during immune/inflammatory stress. The first study assessed hypothalamic kisspeptin gene expression and pituitary LHβ and GnRH receptor expression, in LPS versus restraint animals, to test whether altered gene transcription in LPS animals resulted in lowered LH secretion. The results of this experiment are still pending. The second study tested the LH response to a single bolus of GnRH or kisspeptin. Interestingly, we found equivalent LH increases, to either agonist, in saline versus LPS-treated animals. Collectively, these data indicate that both restraint and immune/inflammatory stress inhibit LH pulse frequency. However, the endogenous LH pulse profiles suggest an additional stressor-specific mechanism that lowers pulsatile LH during immune/inflammatory stress via LPS. Identification of stress pathways that alter endocrine hormones is critical as it allows for the development of therapeutics and medications to treat endocrine disorders. Presentation: Saturday, June 17, 2023

FRI279 Submaximal Loss Of KNDy Neurons Accelerates And Amplifies Pulsatile LH Secretion In Female Rats

Abstract Disclosure: A.C. Campideli-Santana: None. K.S. da Costa Silva: None. R. Araújo-Lopes: None. L.M. Antunes: None. R.E. Szawka: None. Kisspeptin is a powerful stimulator neuropeptide in the hypothalamus-pituitary-gonadal axis and is essential for fertility. Neurons in the arcuate nucleus (ARC) of the hypothalamus that coexpress kisspeptin, neurokinin B, and dynorphin (KNDy) play a crucial role in the control of luteinizing hormone (LH) secretion, but the neuroendocrine mechanisms involved in this regulation are not fully understood. Our previous study demonstrated that the moderate loss of KNDy neurons amplifies the LH surge induced by estradiol in ovariectomized rats (1). The current study aimed to perform a longitudinal evaluation of female rats in which KNDy neurons were ablated to determine the integrated role of this neuronal population in the control of pulsatile LH secretion. Adult female rats underwent a neurochemical ablation of KNDy neurons via intra-ARC stereotaxic injections of neurokinin-3 receptor agonist conjugated with saporin (NK3-SAP), while control animals received blank-saporin (Control). Estrous cyclicity was monitored over 21 days before and after the stereotaxic surgery. Three weeks after surgery, blood samples (10 µL) were withdrawn from the tail tip at 6-minute intervals for 180 minutes on the day of diestrus. Rats were then ovariectomized and the ovaries were processed for histological analysis. Whole blood LH levels were measured by ELISA and brains were immunohistochemically labeled for neurokinin B in the ARC. NK3-SAP rats bearing a submaximal loss of KNDy neurons (between 55-95%; n = 8) displayed irregular estrous cycles. The ovarian weight and the number of corpora lutea and atretic follicles were unaltered, whereas the number of healthy early antral follicles decreased in NK3-SAP rats. The frequency of LH pulses was increased and both pulse amplitude and mean LH levels were higher in NK3-SAP compared with Control rats. Thus, the submaximal loss of KNDy neurons affects the regularity of estrous cyclicity but does not preclude ovulation, while seems to stimulate the maturation of early antral follicles. Notably, KNDy neuron loss accelerates and amplifies LH pulsatile secretion during the rat estrous cycle. These results suggest a new role for KNDy neurons in restraining the frequency and amplitude of LH pulses in ovary-intact animals, with possible implications in the rate of recruitment of early antral follicles. Reference: Campideli-Santana et al., J Neuroendocrinol. 2022 Nov;34(11):e13204. Support: FAPEMIG, CNPq, CAPES, and Pro-reitoria de Pesquisa-UFMG. Presentation: Friday, June 16, 2023

Impact of baseline luteinizing hormone (LH) level and anti-Müllerian hormone (AMH) on pregnancy outcomes in polycystic ovary syndrome (PCOS) patients undergoing IVF/ICSI cycle in antagonist protocol

Introduction: Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder often associated with infertility. There are several factors that could impair the pregnancy outcomes of PCOS patients. The latter endocrine profile along with high baseline luteinizing hormone (LH), high LH/follicle-stimulating hormone ratio, as well as high anti-Müllerian hormone (AMH) levels appear detrimental for patients undergoing In vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). The objective of this study is to evaluate the impact of baseline LH and AMH levels on pregnancy outcomes in PCOS patients performing IVF/ICSI cycles in an antagonist protocol. Materials and methods: The current investigation is a retrospective cohort study conducted at IVF Tiziri Center in Algiers on PCOS patients undergoing IVF/ICSI from January 2017 to March 2021. According to the Rotterdam criteria, patients aged between 23 and 42 years were divided into 2 groups: group 1 (G1) =91 patients; group 2 (G2) =100 all patients had an LH level &gt;10 IU/L. Both groups had AMH levels &gt;5 ng/mL. Controlled ovarian stimulation was done with a fixed antagonist protocol, as for the ovulation it was triggered with human chorionic gonadotropin or gonadotropin-releasing hormone agonist. Fresh and frozen embryo transfers were, thereafter, evaluated. Concerning the statistical analysis, it was done with SPSS software. Results: The mean LH level was 15.16±7.08 for G1 versus 5.7±1.86 for G2 (P=10−6), mean LH/follicle-stimulating hormone ratio 2.71±1.36 in G1 versus 1.05±0.44 (P=10−6) and mean AMH level was 9.27±3.90 for G1 versus 7.53±2.76 for G2 with (P&lt;10−3). The mean follicles over 11 mm in diameter, estradiol (E2) level, and progesterone level on the trigger day did not differ significantly between G1 and G2. Also, the mean of oocytes and metaphase II oocytes retrieved fertilization, and blastulation rate were similar for both groups. The pregnancy rate for G1 was 40.6% versus 54% (P&gt;0.05); the ongoing pregnancy rate (OPR) was 28.6% in G1 versus 47% in G2 (P=0.078); and the miscarriage rate was 32.4% for G1 versus 12.9 for G2 (P=0.072). A univariate analysis was performed with all confounder factors supposed to impact pregnancy outcomes: AMH level did not impact pregnancy outcomes, whereas LH level &lt;11 IU/L has been determined as a threshold impacting positively on OPR with odds ratio=2.01 (95% CI: 1.05–3.87). After a multivariate logistic regression including con-founders, 3 factors significantly influence OPR: LH, E2, and infertility with known etiology. Conclusion: The high baseline LH level impairs pregnancy outcomes in PCOs patients undergoing an antagonist protocol IVF/ICSI cycle. The identification of PCOS patients by their endocrine profile enables the establishment of a prognosis for council patients and a plan for an individualized therapeutic strategy.

Assessment of pubertal onset and disorders of puberty in Indian children and youth with type-1 diabetes.

Disorders of pubertal development are enlisted as associated conditions in children and adolescents with type-1 diabetes (T1D). We conducted this study with objective (1) To estimate the median age at onset of puberty and luteinizing hormone (LH) and sex-steroid concentrations in Indian adolescents with T1D and (2) To assess the impact of puberty on glycemic control and insulin resistance (IR). This cross-sectional study included 399 children and youth aged 6-23 years with T1D. Demographic, anthropometric, biochemical and pelvic ultrasound data were collected using standard protocols. IR was calculated using estimated glucose disposal rate and puberty was assessed using Tanner staging. Median age at onset of thelarche, pubarche and menarche were 11.3, 11.4 and 12.8 years in girls and that of gonadarche and pubarche were 10.6 and 12.7 years for boys. The mean LH and sex-steroid concentrations of subjects with T1D were similar to healthy subjects at each stage of puberty. The cut-offs of LH and sex-steroids derived from healthy Indian children yielded high sensitivity and specificity in determining pubertal onset. The prevalence of precocity, delayed puberty, ovarian cysts and polycystic ovaries was 0.9 , 5.1, 5.1 and 8.6 %, respectively. Glycaemic control and insulin sensitivity was poor in pubertal subjects. The age at onset of puberty, LH, and sex-steroid concentrations in subjects with T1D were like otherwise healthy Indian children with poor glycemic control and IR in pubertal subjects. Although most complications of T1D are associated with poor glycemic control, pubertal disorders were significantly low despite the less-than-optimal glycemic control.

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

Corticotropin releasing factor-1 receptor antagonism associated with favorable outcomes of male reproductive health biochemical parameters.

Disruption in androgen profiles and testicular adrenal rest tumors in males with congenital adrenal hyperplasia (CAH) can negatively affect sexual activity and fertility. Adrenal hyperandrogenism suppresses gonadotropin secretion and testicular adrenal rest tumors (TARTS), despite being noncancerous lesions, cause obstructive azoospermia and impaired testosterone (T) production. Circulating T in men with uncontrolled CAH is often predominantly adrenal in origin, which is reflected in high androstenedione/testosterone ratios (A4/T). Therefore, decreased luteinizing hormone (LH) levels and an increased A4/T are markers of impaired fertility in these individuals. Oral tildacerfont 200 to 1000 mg once daily (QD) (n=10) or 100 to 200 mg twice daily (n=9 and 7) for 2 weeks (Study 201), and 400 mg QD (n=11) for 12 weeks (Study 202). Outcomes measured changes from baseline in A4, T, A4/T, and LH. Mean T levels increased in Study 201 from 375.5 ng/dL to 390.5 ng/dL at week 2 (n=9), 485.4 ng/dL at week 4 (n=4) and 420.7 ng/dL at week 6 (n=4). In Study 202, T levels fluctuated in the normal range from 448.4 ng/dL at baseline to 412.0 ng/dL at week 12. Mean LH levels increased in Study 201 from 0.68 IU/L to 1.59 IU/L at week 2 (n=10), 1.62 IU/L at week 4 (n=5) and 0.85 IU/L at week 6 (n=4). In Study 202, mean LH levels increased from 0.44 IU/L at baseline to 0.87 IU/L at week 12. Mean A4/T decreased across both studies. In Study 201, the mean A4/T changed from a baseline of 1.28 to 0.59 at week 2 (n=9), 0.87 at week 4 (n=4), and 1.03 at week 6 (n=4). In Study 202, the A4/T decreased from baseline of 2.44 to 0.68 at week 12. Four men were hypogonadal at baseline; all experienced improved A4/T and 3/4 (75%) reached levels <1. Tildacerfont treatment demonstrated clinically meaningful reductions in A4 levels, and A4/T with concomitant increased LH levels indicating increased testicular T production. The data suggests improvement in hypothalamic-pituitary-gonadal axis function, but more data is required to confirm favorable male reproductive health outcomes.

Abstract #1375340: Correlation of Follicle Stimulating Hormone and Luteinizing Hormone Levels with HbA1c in Post-Menopausal Females

Menopause in female is a normal physiological process however, it is also a well-established risk factor for CVD and osteoporosis. Few studies suggest that low FSH is associated with adiposity and diabetes in post-menopausal women. Aim of this study was to explore correlations of follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels with HbA1c level in post-menopausal females. The study was conducted at of Dr. S.N. Medical College, Jodhpur, India. Otherwise healthy postmenopausal women attending hospital for preventive health checkup were included in study. Postmenopausal status was confirmed by the absence of menses for at least 12 months. Known case of T2D/T1D, subjects on hormone replacement therapy, acutely sick were excluded from study. The baseline fasting blood samples were collected. Samples was assayed for FSH, LH, HbA1c and standard biochemical profile (glucose, lipid profile, Live function tests and renal function tests) using standard methods. The participants were classified as having diabetes or prediabetes using ADA criterias. Total 229 postmenopausal women were enrolled. Mean age of subjects was 63.2±9.9 years. Mean Hba1c was 6.2±1.4 gm% and subjects were further divided in to non-diabetes (68/229), prediabetes (104/229) and diabetes group (57/229) according to Hba1c levels. Mean LH was 25.8±15.6, FSH was 61.1±30.1 and LH/FSH ratio was 0.47±0.35. Mean LH was 31.4±15.8, 26.0±16.3, 19.5±12.6 in non-diabetes, prediabetes and diabetes group respectively. Mean FSH was 75.9±29.3, 59.6±27.6, 46.9±29.0 in non-diabetes, prediabetes and diabetes group respectively. The LH and FSH values in non-diabetes and diabetes group were statistically significant (p<0.01). In this study low FSH and LH levels were associated with higher level of HbA1c and increased risk of T2D.

Clomiphene citrate: A potential alternative for testosterone therapy in hypogonadal males.

Hypogonadism is a worldwide problem among men causing sexual, physical and mental problems. Testosterone therapy is the first-choice treatment for male hypogonadism, with several side effects, that is, subfertility. Clomiphene citrate (CC) is an alternative off-label therapy for a certain group of hypogonadal males, especially for those with an active or future child wish. There is scarce literature in usage of CC for men with hypogonadism. The aim of this retrospective study was to evaluate the effectiveness and safety of CC for hypogonadal males. In this single-centre study, men treated with CC for hypogonadism were evaluated retrospectively. Primary outcome was hormonal evaluation including total testosterone (TT), free testosterone (FT), luteinizing hormone (LH) and follicle stimulating hormone (FSH). Secondary outcomes were hypogonadal symptoms, metabolic and lipid parameters, haemoglobin (Hb), haematocrit (Ht), prostate specific antigen (PSA), side effects, the effect of a trial without medication and potential predictors for biochemical and clinical response. In total, 153 hypogonadal men were treated with CC. Mean TT, FT, LH and FSH increased during treatment. TT increased from 9 to 16 nmol/L, with a biochemical increase in 89% of the patients. In patients who continued CC treatment, an increased level of TT persisted after 8 years of treatment. With CC treatment, 74% of the patients experienced hypogonadal symptom improvement. LH at the lower normal range before CC treatment was predictive for better TT response. During CC therapy, few side effects were reported and no clinical important changes in PSA, Hb and Ht were found. Clomiphene citrate is an effective therapy on short and long term, improving both clinical symptoms and biochemical markers of male hypogonadism with few side effects and good safety aspects.