Mean LH Levels Research Articles

P-666 Comparison of IVF outcomes in women undergoing IVF stimulation using progestin primed ovarian stimulation (PPOS) Vs GnRH antagonist (GnRH-Antag) protocol

Abstract Study question Can PPOS achieve improved patient & provider satisfaction while simultaneously blocking LH-surge, maintaining similar IVF outcomes like oocyte/embryo yield/clinical pregnancy rates(CPR) when compared to GnRH-Antag? Summary answer PPOS can achieve similar IVF outcomes as compared to GnRH-Antag protocol and is a patient friendly, cost-effective choice for ovarian stimulation without affecting oocyte/embryo yield/CPR. What is known already GnRH-Antag protocol is among the most widely used methods, as it blocks LH surge & significantly lowers the risk of OHSS [Lambalk CB,2017]. However, this protocol has disadvantages like frequent monitoring, dose adjustment, high therapeutic cost and multiple injections[Ata B,2015]. PPOS successfully prevents ovulation and provides comparable IVFoutcomes (oocyte yield, blastocyst/euploid embryo rates, CPR) along with patient cost savings and increased convenience across all infertility diagnoses [Annalyn M.Welp, 2023]. It should be noted that most studies are non-randomized and comprise low to moderate-quality evidence. However, the consistency of the findings across different studies from different centres provides assurance in effect estimates. Study design, size, duration A retrospective cohort study conducted at a tertiary care fertility unit. Data for 449 women aged 21 to 40 years undergoing IVF stimulation using PPOS (n = 226) and GnRH-Antag (n = 223) protocol was retrieved from the hospital’s electronic database and analysed for a period between September 2022 to October 2023. Study population consisted of women undergoing freeze-all strategy either for lowering the risk of OHSS (hyper-responders) or for embryo pooling in view of very low ovarian reserve. Participants/materials, setting, methods For both groups either rFSH/rLH/HMG were used & doses were indivisualised based on patient’s age, BMI & ovarian reserve. PPOS group received twice daily dose of medroxyprogesterone acetate tablet(10 mg)/dydrogesterone(10 mg) from Day2 of cycle. GnRH-Antag group received Inj.Cetrorelix acetate (0.25 mg) daily as flexible protocol. Final oocyte maturation was induced with Triptorelin(0.2mg)/Choriogonadotropin alfa(250mcg)/both, once 2-3follicles reached 18-20mm. Transvaginal oocyte pick-up was performed 35-36h later. Women in PPOS group underwent freeze-all strategy followed by Frozen-Thawed Embryo-transfer. Main results and the role of chance Normally distributed continuous variables were compared using independent t-test, and categorical variables were compared by χ2 and Fisher’s exact test, where appropriate. Patient demographics, clinical/ovarian stimulation characteristics, embryological and IVF outcomes were compared for two groups. Patient demographics {Age(p-value:0.08), BMI(p-value:0.06), duration(p-value:0.09)/type(p-value:0.76) of infertlity}, total gonadotropin dose (p-value: 0.39) and duration of ovarian stimulation (p-value: 0.81) were comparable between the two groups. Though the mean LH level on trigger day was slightly higher in PPOS group (5.53 IU/L) than GnRH-Antag group (3.24 IU/L) (p-value: 0.02), statistically significant difference was not seen in the oocyte yield (p-value: 0.18), M-II oocytes (p-value: 0.17) & fertilization rate (p-value: 0.23) in the two groups. The number of Day3 (p-value: 0.65), Day5 (p-value: 0.07) & Day6 (p-value: 0.08) embryos available in the two groups were also comparable. IVF outcomes in terms of positive B-HCG result (p-value: 0.68), clinical pregnancy rate (p-value: 0.61) & miscarriage rate (p-value: 0.27) showed no statistically significant difference in the two groups. Thus, we conclude that PPOS can achieve similar IVF outcomes as compared to the GnRH-Antag protocol, while at the same time proving to be patient friendly by reducing the need of multiple injection pricks, injection cost and the frequency of hospital visits. Limitations, reasons for caution This is a retrospective study and hence randomized comparison was not possible. PPOS can be used only when freeze-all strategy is planned, fresh embryo-transfer cannot be considered if need arises depending upon embryo quality. IVF service providers have to depend on patient compliance for using oral progestins to block LH-surge. Wider implications of the findings Study findings suggest that PPOS is a patient friendly and equally effective and safe IVF stimulation protocol when compared to GnRH-Antagonist. More RCT will be useful for investigating different aspects of PPOS (it’s utility for women with different ovarian reserve functions, conventional versus flexible approach, optimal dose of each progestin). Trial registration number Not applicable

P-755 Effect of female age and specific cycle characteristics on success in modified natural frozen-thawed single euploid embryo transfer cycles (mNC/FET)

Abstract Study question Does the age of woman and specific cycle characteristics impact the success of modified natural frozen-thawed single euploid embryo transfer cycles (mNC/FET)? Summary answer The success rate of mNC/FET cycles is similar in all age groups. As the LH level increased ( above 23.5ng/dL) the success was significantly increased. What is known already The success of frozen embryo transfer can depend on various factors such as the patient’s age, the number of transferred embryos, the duration of embryo culture, and the method used to prepare the endometrium (Holschbach et al. 2023). However, there is currently insufficient evidence in the literature about the effect of age and cycle characteristics on the effectiveness of mNC/FET cycles in patients who have received single top and good quality euploid embryos. Study design, size, duration This study, conducted at Istanbul Memorial Hospital, ART and Reproductive Genetics Center, is a retrospective, single-centre analysis of 583 cycles of euploid mNC/FET in women aged 22-44 years. The study period was from July 2017 to January 2024. The cycles were divided into two groups based on the age of the women. Group A (n = 408) included women under the age of 38, while Group B (n = 175) included women aged 38 or above. Participants/materials, setting, methods The study investigated success predictors in mNC/FET with top and good-quality embryos. Excluded were women with recurrent pregnancy losses, recurrent implantation failure, high BMI (>35kg/m2), uterine/endometrial problems, severe male infertility, and autoimmune diseases. The mNC/FET success was a pregnancy lasting 12 weeks or more. The analysis included patient demographics and cycle outcomes. Chi-square, t-tests, and Mann-Whitney U tests assessed variables with a p-value < 0.005, indicating statistical significance. Main results and the role of chance No significant demographic differences were observed between the two groups, except for AMH levels. AMH levels were significantly higher in younger group than older group (3.02 ng/ml vs. 1.96 ng/ml, p = 0.000). The mNC/FET success rates were similar, with 76% in the younger group and 73% in the older group. Estradiol levels the day before trigger day were significantly higher in older group (278.02 pg/ml vs. 237.84pg/ml, p = 0.028). Both groups had similar mean LH levels (32.50 IU/L and 32.41 IU/L, respectively), estradiol levels and endometrial thickness on the trigger day. In the younger group, LH rise was observed later, follicular phase was longer, and embryo transfer day was later than the older group (p = 0.001). Univariate regression analysis revealed that increased LH level was an important independent variable in achieving mNC/FET success, with a 1.001-fold increase (p = 0.043). The LH levels of 23.5 IU/L or higher predicted high mNC/FET success with 60% sensitivity and 43% specificity based on the Receiver Operating Characteristic curve. (AUC: 0.606, %95 CI 0.540-0.671, p = 0.002). Prolonged infertility duration and previous ET failures reduced mNC/FET success by 0.889 and 0.544-fold, respectively (p = 0.002, p < 0.001). Higher β-hCG levels on days 14 and 16 increased mNC/FET success by 1.038-fold and 1.008-fold, respectively (p < 0.001). Limitations, reasons for caution Our study has limitations as it was retrospective. Prospective randomized studies are needed to evaluate the differences between the young (age <38) and old patient groups (age ≥38) based on mNC/FET success. The association between LH rise and mNC/FET success needs verification with a larger sample. Wider implications of the findings The mNC/FET success rates are similar in young (age <38) and old patient groups (age ≥38). However, prolonged infertility duration and previous failed ET cycles decreased the success of mNC/FET. Additionally, It is important to schedule trigger time after the optimum LH rise, which is 23.5 IU/L or higher. Trial registration number not applicable

Prevalence Of Skin Manifestations In Patients With Polycystic Ovarian Syndrome

Objective: This study aimed to determine the prevalence of cutaneous manifestations in PCOS patients. Method: This cross-sectional study was conducted in the dermatology department, Combined Military Hospital, Peshawar, from 1st January 2021 to 30th June 2021. After obtaining ethical approval from the hospital's ethical and research committee, a total of 50 patients aged 15 to 45 years were included in the study fulfilling the inclusion criteria. Written informed consent was taken from each patient. All patients underwent general physical, systemic and dermatological examination. Venous blood sampling was done after overnight fasting for hormonal assays (TSH, LH, FSH, prolactin and testosterone), glucose, and lipid levels. An ultrasound scan was done for the detection of ovarian cysts. Results: Mean age was 26.96 years ± 5.42 SD. The mean BMI was 26.04 Kg/m2 ± 5.10 SD. Mean FSH and LH levels were 6.86 IU/L ± 5.41 SD and 12.11 IU/L ± 8.75 SD respectively. Among them, 88% were married, 36% had menstrual irregularities and 72% had infertility. Hirsutism, acne, acanthosis nigricans, and androgenic alopecia were noted in 72%, 54%, 42%, and 20% of the patients respectively. Conclusion: Cutaneous manifestations can play an important role in the diagnosis of PCOS and also contribute an important portion of the symptoms experienced by patients suffering from PCOS. Keywords: PCOS, Skin manifestations, Hyperandrogenism, Hirsutism, Acne.

Effects of rankl inhibitor on hormonal parameters in letrozole induced rat model of polycystic ovary syndrome.

Objective: To study the effect of RANKL inhibitor on serum FSH, LH and testosterone level in Letrozole induced rat model of PCOS. Study Design: Experimental Animal study. Setting: Thirty adult female Wistar Albino rats of 6-8 weeks of age with a weight range of 120–150 gm were recruited from the animal house of Baqai Medical University. Period: February 2022 till August 2022. Material & Methods: The rats were randomly divided into A, B, and C groups and each contained 10 rats. Group A was considered ass control while Group B served as Letrozole induced PCOS group and Group C served as RANKL inhibitor group. Letrozole dissolved in 0.1% CMC and was given at a dose of 1 mg/kg orally for 21 days to induce PCOS. RANKL inhibitor (Denosumab) was given 0.25 mg/kg body weight subcutaneously for 4 weeks after the induction of PCOS i.e from 21 days to 50th days. At the end of the experiment, blood was taken from intra cardiac puncture and analyzed through ELISA. Results: Mean serum level of LH and testosterone level were significantly higher (P<0.001) in Letrozole-treated PCOs group which was significantly decreased (P<0.001) in Denosumab treated animals but significantly higher (P<0.05) than control group. Whereas, mean serum FSH level was significantly (P<0.01) lower in the Letrozole-treated group which was significantly increased in Denosumab treated animals but significantly lower (P<0.05) than the control group. Conclusion: RANKL inhibitor could prevent the hormonal imbalance in letrozole treated PCOs in albino rats.

SAT286 Effect of Tildacerfont On Gonadotropins And Testicular Steroidogenesis In Men With Classic 21-hydroxylase Deficiency

Abstract Disclosure: R. Charlton: Employee; Self; Spruce Biosciences, Inc. A. Khattab: Consulting Fee; Self; Antares Pharma, Inc. R.J. Auchus: Grant Recipient; Self; Spruce Biosciences, Inc. Research Investigator; Self; Spruce Biosciences, Inc. Impaired testicular function is a common comorbidity of classic 21-hydroxylase deficiency (21OHD). Testicular adrenal rest tumors (TART) can disrupt testicular architecture and cause primary hypogonadism. Both supraphysiologic glucocorticoid doses and adrenal-derived androgens and estrogens can suppress gonadotropins and cause secondary hypogonadism. We conducted a post-hoc analysis of gonadal axis function in men with 21OHD participating in SPR001-201 and SPR001-202, two phase 2 studies of tildacerfont, a once-daily oral corticotrophin-releasing factor receptor 1 (CRFR1) antagonist, designed to reduce ACTH and adrenal steroid production, including androstenedione (A4). Study 201 included 3 cohorts: Cohort A, 5 men who received tildacerfont in a 6-week dose escalation - 2 weeks at 200 mg QD, 2 weeks at 600 mg QD, and 2 weeks at 1000 mg QD without washout between doses; Cohort B, 1 man who received tildacerfont for 2 weeks at 200 mg BID; and Cohort C, 4 men who received tildacerfont for 2 weeks at 100 mg BID. Study 202 included 4 men who received tildacerfont 400 mg QD for 12 weeks. Participants continued their previously prescribed glucocorticoid ± mineralocorticoid replacement without dose adjustments. Geometric means were used to describe hormone levels due to the small participant numbers. Mean and median ages were 46/32y in Study 201 and 32/35y in Study 202 (range 18-54 in both studies). All participants reported race as white except one who identified as white and Asian. Four reported ethnicity as Hispanic or Latino. At baseline, testosterone (T) levels were normal except for four 201 participants that had T <300 ng/dL. At baseline, 6/11 (55%) had suppressed LH (including the four with low T), and 11/14 (79%) had A4/T ratios >1, indicating a predominantly adrenal origin of T. In Study 201, 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, mean T levels were 448.4 ng/dL at baseline and 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) (normal 1.8-8.6IU/L). In Study 202, mean LH levels increased from 0.44 IU/L at baseline to 0.87 IU/L at week 12. Of the four men who had low T and suppressed LH (hypogonadotropic hypogonadism), LH rose in all 4 and normalized in 3/4 (75%). Mean A4/T ratios decreased across both studies. In Study 201, the mean declined 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 ratio decreased from baseline of 2.44 to 0.68 at week 12. All the hypogonadal men experienced reductions in A4/T ratios, and 3/4 (75%) achieved A4/T <1. This post-hoc analysis highlights that gonadal axis disruption is common in men with 21OHD and suggests that modulation of adrenal androgen production via CRFR1 antagonism may improve testicular function. Presentation: Saturday, June 17, 2023

The role of anti-Mullerian hormone and other correlates in patients with polycystic ovary syndrome

ABSTRACT Background Anti-Müllerian hormone (AMH) has recently emerged as a promising biomarker for the detection of polycystic ovarian morphology. In polycystic ovary syndrome (PCOS), an elevated level of AMH has been suggested to add value to the Rotterdam criteria in cases of diagnostic uncertainty. In this study, we evaluated the correlation between AMH and PCOS, and the potential role of AMH in PCOS diagnosis. Methods A case-control study was performed on a total of 200 females, 100 of which were diagnosed with PCOS as per Rotterdam revised criteria (2003) and 100 as the control (non-PCOS group). Patient medical records were therefore retrieved for clinical, biochemical and ultrasound markers for PCOS diagnosis. Sensitivity, specificity, area under receiver operating characteristic (AUROC) curve, and multivariate linear regression models were applied to analyze our data. Results Mean serum levels of LH and AMH, and LH/FSH ratio were significantly different between compared groups. In the PCOS group, the mean serum AMH level was 6.78 ng/mL and LH/FSH ratio was 1.53 while those of controls were 2.73 ng/mL and 0.53, respectively (p < .001). The most suitable compromise between 81% specificity and 79% sensitivity was obtained with a cutoff value of 3.75 ng/mL (26.78 pmol/L) serum AMH concentration for PCOS prediction, with an AUROC curve of 0.9691. Conclusion Serum AMH cutoff level of 3.75 ng/mL was identified as a convenient gauge for the prediction of PCOS and an adjuvant to the Rotterdam criteria.

Gonadotropin-Releasing Hormone Studies to Assess Pubertal Deficiencies in Prader-Willi Syndrome

Patients with Prader-Willi Syndrome (PWS) are known to have variable pubertal development with most having delayed or incomplete puberty. This study aims to evaluate the activity of the Hypothalamic-Pituitary-Gonadal (HPG) axis in individuals with PWS. Thirty-five patients (18 males, 17 females) ages ranging from 3 to 38 years with genetically verified PWS underwent GnRH stimulation testing using gonadorelin along with clinical evaluations. Of patients in the typical age range for initiation of puberty, six had LH peak &gt;5IU/L, indicating activation, but the remaining six had variable peaks, indicating lack of, or indeterminate activation. In some females, physical examination noted breast development to Tanner stage II to III, despite apparent lack of HPG axis activation, while in some males testicular volumes were smaller than expected despite apparent activation of the HPG axis. We studied genotype differences (deletion vs UPD) with respect to HPG axis activation. Mean LH levels were nearly double for PWS females with UPD compared to PWS females with deletion. Both peak LH and peak FSH levels appear to be higher in males with deletion compared to UPD. Inconsistency between physical examination and stimulation testing results and the high variability in the age at activation of the HPG axis accentuates the need for the evaluation of pubertal staging in patients with PWS, and consideration of sex-steroid supplementation.

PSAT072 Clinical Features of Patients with 21-Hydroxylase Deficiency and Testicular Adrenal Rest Tumors

Abstract Testicular adrenal rest tumors (TARTs) are one of the most frequent comorbidities in males with 21-hydroxylase deficiency (CAH) and different prevalence rates have been reported. TART still present difficult control and frequently lead to infertility. Since it is also observed in patients with adequate hormonal control, its predictors factors are not yet well-known. Objective To evaluate TARTs prevalence in a significant CAH male cohort and to correlate with the long-term predictors and their impact on patients’ gonadal function. Patients and methods: Data of 53 males (24 salt wasters, 10 non-classical) were retrospectively evaluated. Hormonal control quality, assessed through long-term (last 5 years) mean ACTH, 17OHP, androstenedione (D4), testosterone (T), FSH, LH, inhibin B and renin levels were analyzed as well as sperm analysis. Mean glucocorticoid doses were evaluated and converted into cortisol equivalent. Genotypes, determined through entire CYP21A2 sequencing, were classified according to predicted impairment on residual enzymatic activity: null and severe groups. Data of patients with/without TARTs by scrotal ultrasound (US) were compared. Student's t and Chi-square tests were used. Results TARTs were observed in 18 (42%) of classical patients, all of them presented with bilateral lesions. None of the non-classical males presented with TART. The mean age of TART diagnosis was 21.32±5.2 years, the youngest patient with TART was 12y6mo old. Ten patients with TART underwent sperm analysis and all of them presented with altered results, while 3/10 patients without TART presented with abnormal sperm count. On the other hand, only 4 (22%) TART patients developed hypergonadotropic hypogonadism. Despite late diagnosis of the SV form in our cohort, the frequency of TARTs was significantly higher in SW patients (54% vs 37%); interestingly, similar frequencies of genotype groups were observed in patient with/without TARTs. Mean glucocorticoid doses did not differ in patients with and without TARTs as well as fludrocortisone doses in SW patients with and without TARTs (p&amp;gt;0.05 for both). Mean 17OHP, D4 and T levels did not differ in both patient groups; mean ACTH levels were 305.3+/-282 vs. 103.2+/-81.9pg/ml in patients with and without TART, respectively (p&amp;gt;0.05). Mean renin levels were significantly higher in patients with TARTs than in those without (152.4+/-116.8 vs. 93.8+/-96.4mIU/ml). Testicular function parameters (mean inhibin B, FSH, LH levels and frequency of abnormal sperm analysis) were worse in the TART group vs without TART (p&amp;lt;0.05). Conclusion in our series, a high frequency of TARTs was observed, leading to male fertility impairment. An additive renin stimulatory effect, instead of long-term hormonal control or genotype, seemed to be the major role in TARTs development. Presentation: Saturday, June 11, 2022 1:00 p.m. - 3:00 p.m.

Reproductive Effects of Medicinal Plant (Azadirachta indica) Used as Forage and for Ethnoveterinary Practices: New Insights from Animal Models

In some African and Asian countries, Azadirachta indica (AI) has been fed to livestock for decades and traditionally used to treat certain animal and human diseases. Recently, there are suspicions that the plant may possess anti-reproductive properties and concerns that the continued use of AI as forage or for folkloric medicine may detrimentally affect reproduction in the subjects. To address these challenges, this work determined the reproductive and fertility effects of a methanolic seed extract of AI (MSEAI) using adult female albino rats (AFARs) as an experimental model. Sixty-four AFARs were randomly assigned into four groups (A–D) of sixteen rats each. Group A was the control while groups B, C and D were treated daily with 50, 100 and 200 mg/kg of MSEAI respectively, for 28 consecutive days via oral gavage. Blood samples were collected for hormonal and biochemistry assays. Ovarian samples from the experimental rats were harvested for histopathological studies. Thereafter, the remaining experimental rats were bred, and certain fertility indices determined. The mean serum FSH and LH levels were significantly decreased (p ≤ 0.05) in the 100 and 200 mg/kg groups. The histopathological studies revealed massive follicular degeneration in the 100 and 200 mg/kg treatment groups. The fertility indices indicated that the post-implantation survival index was 100% in the control and 0% in the 200 mg/kg treatment group. No abortion occurred in the control and 50 mg/kg groups, but 25% and 100% of the pregnant does aborted in the 100 and 200 mg/kg treatment groups, respectively. Considering that high doses (100 mg/kg and 200 mg/kg) of MSEAI had significant anti-reproductive and antifertility properties, the use of AI as forage or for ethnoveterinary medicine in breeding females may adversely affect their reproductive potentials. However, the anti-reproductive and antifertility effects could be utilized in rodent depopulation programs in animal agriculture and as a contraceptive to limit the proliferation of stray dogs, known to be reservoirs of the rabies virus in developing countries. Moreover, the MSEAI could be further refined for human use as an effective, cheap, eco-friendly and acceptable alternative to synthetic/modern contraceptives, the use of which is limited in developing nations due to superstitious beliefs and their multiple side effects.

A Case-Control Study of the Luteinizing Hormone Level in Luteinizing Hormone Receptor Gene (rs2293275) Polymorphism in Polycystic Ovarian Syndrome Females

Background: Polycystic ovary syndrome (PCOS) is a common heterogeneous disorder characterized by chronic anovulation, infertility, polycystic ovaries, and hyperandrogenic signs. Objective: The aim of this study was to determine the association of luteinizing hormone/chorionic gonadotropin hormone receptor LHCGR polymorphism (rs2293275) with oligomenorrhea, amenorrhea, hirsutism, acne, infertility, LH, LH/FSH ratio, and body mass index (BMI) among PCOS females. Methods: This genetic case-control study recruited 55 PCOS and 55 control females, diagnosed based on the Rotterdam criteria. LH and FSH were measured by the Roche cobas c 502 automated analyzer. Genotypic analysis was carried out using the polymerase chain reaction-restriction fragment length polymorphism and restriction endonuclease digestion. Results: BMI was higher for PCOS patients (28.5 ± 6.59) compared to controls (25.1 ± 5.77), and ovulatory dysfunction was seen among 90% of PCOS females. Oligomenorrhea was common in PCOS (73%), and hirsutism and acne were detected in PCOS (80% and 40%; respectively). LH ≥10 were recoded among 51%, while LH/FSH ≥1.5 was recorded among 33% PCOS females. There is a statistical difference between rs2293275 polymorphism in the AG genotype between PCOS patients and controls. PCOS patients have a significantly higher mean LH level compared to controls (8.36 ± 4.86 and 5.67 ± 2.51, respectively) and showed higher LH/FSH value (1.46 ± 0.81) compared to (0.87 ± 0.30) controls. GG and AG genotypes of LHCGR showed statistically significant higher LH (8.22 ± 4.11; 9.02 ± 3.87) and LH/FSH values (1.57 ± 0.56; 1.64 ± 0.89) compared to controls. Conclusion: LHCGR (rs2293275) GA and GG genetic variants could modulate the hormonal levels of PCOS LH levels and the LH/FSH ratio and associated with hirsutism, oligomenorrhea, BMI, and LH/FSH ratio as risk factors.

Comparison of the sex hormones\u2019 serum level in women with recurrent aphthous stomatitis and healthy population: a cross-sectional study

BackgroundIn this study, we aimed to evaluate the sex hormonal serum level in patients with recurrent aphthous stomatitis and compare them with healthy participants.MethodsThis cross-sectional study was done on patients with recurrent aphthous stomatitis who had referred to Shiraz Dental Faculty, Oral and Maxillofacial Medicine Department during 2018–2019. The non -menopause women with recurrence of at least 3 lesions per year were enrolled in this study. The mean serum level of FSH, LH, PRL (prolactin), testosterone, DHT (Dihydrotestosterone), DHEA-S (Dehydroepiandrosterone sulfate), estradiol and progesterone of 30 participants in each group of case and control were measured and compared. The data were analyzed by SPSS version 18 and independent T-test, Mann–Whitney U test, Spearman’s correlation coefficient test, Chi-square test and Fisher’s test.ResultsThe mean serum level of DHEA-S in patients with recurrent aphthous stomatitis (RAS) was significantly lower than the control group (p value = 0.02). In addition to DHEA-S, the mean serum level of testosterone was lower in the evaluation group although this difference was not significant (p value = 0.057). Considering the effect of age on the mean serum level of sex hormones, our results revealed that only DHEA-S mean serum level was decreased by increasing the age of participants in patients with RAS (p value = 0.018). The number of participants with abnormal range of testosterone (p value < 0.0001) and progesterone (p value = 0.037) serum level was significantly more in patients with RAS. The frequency of RAS in a year did not show a significant relationship with the serum level of the evaluated hormones.ConclusionThe patients with RAS had a lower serum level of DHEA-S. The mean serum level of testosterone and progesterone was significantly abnormal in RAS patients.

Follicular-Stimulating Hormone, Luteinizing Hormone, and Prolactin Serum Level in Patients with Oral Lichen Planus in Comparison to Healthy Population.

Materials and Methods In this cross-sectional case control study, the serum level of LH, FSH, and prolactin of 40 women with lichen planus who have been referred to Shiraz Dental Faculty, Oral and Maxillofacial Disease Department during 2018-2019 has been evaluated in comparison to 40 healthy controls. Data were analyzed by SPSS version 18. Two-way ANOVA and Mann–Whitney test were used for data analysis. Results The mean serum level of FSH and LH was significantly higher in OLP patients while this difference was not reported for prolactin. Only FSH mean serum level was significantly higher in nonmenopausal OLP patients. The distribution of prolactin and FSH hormones' serum level was in normal range. Conclusions The high serum level of FSH and LH can affect OLP pathogenesis by estrogen and progesterone modulation.

P–710 The possible relation between kisspeptin and apoptosis on granulosa cells of PCOS patients

Abstract Study question Does kisspeptin administration affect the apoptotic factors of granulosa cells of Polycystic Ovary Syndrome (PCOS) patients? Summary answer Kisspeptin administration significantly increased apoptosis related factors. What is known already Kisspeptin plays a critical role in central gonadotropin secretion by acting on GnRH neurons in the hypothalamus. Kisspeptin receptors are also expressed on the ovaries. Kisspeptin–54 has recently been known as a prominent agent in oocyte maturation. Although there are many studies on the possible roles of kisspeptin administration in vivo on hypothalamic gonadotropin secretion and ovarian function and the possible roles of these effects in the pathogenesis of PCOS, there is not enough information about the effect of in vitro application of kisspeptin on ovarian function and conditions that lead to the development of PCOS. Study design, size, duration This basic research study was an in vitro experimental approach involving the use of granulosa cell from a PCOS and normal cases between July to December in 2020. 32 women were included in both control and PCOS groups. Participants/materials, setting, methods Women with (n = 16) and without (n = 16) PCOS were included in the study. Granulosa cells, follicular fluids and blood samples were collected on oocyte pick-up day. Cells were isolated from follicular fluids. Kisspeptin–54 levels were determined by ELISA in serum and follicle fluids. Also, FSH and LH levels were determined in serum. Kisspeptin–54 administrated in vitro to the half of the samples. Fluorescence spectrophotometer and laser scanning confocal microscope were used to evaluate calcium concentrations Main results and the role of chance The serum kisspeptin level was found significantly higher in PCOS patients compared to the control group (p &amp;lt; 0.05). Also, the level of kisspeptin in follicular fluid was statistically significantly higher in PCOS patients compared to the control group (p &amp;lt; 0.05). The mean serum FSH level of PCOS patients was found to be significantly lower than the control group (p &amp;lt; 0.05). Furthermore, the mean serum LH level of PCOS patients was significantly higher than control groups (p &amp;lt; 0.05). The intracellular calcium concentration in granulosa cells obtained from PCOS patients was significantly lower than the control group (p &amp;lt; 0.05). Limitations, reasons for caution The KISS1, KISS1R, BCL–2, CASPASE–3, FSHR, LHCGR, STAR, ESR1, ESR2, INHBA gene expression levels of apoptotic process is still under analyses. The protein expression analyses of these genes may strength the results. Wider implications of the findings: Using kisspeptin antagonists may be supposed as an additional treatment for patients with PCOS. Trial registration number OMU KAEK 2019 /724

The circulating alarin level was elevated in infertile women with poor ovarian reserve

Objective We aimed to reveal the association of serum alarin level with POR status of the infertile women in the present study. Methods The eligibility criteria for this prospective cross-sectional study included a total of 92 infertile women attending the Hitit University Hospital, and all participant women were categorized into two main groups of ovarian reserve: (i) Poor ovarian reserve (POR) group (n = 40) based on ESHRE consensus and (ii) Control group (NOR) (n = 52). Results The mean adjusted-ages and BMI values of the NOR and POR groups were statistically comparable (p = .057 and p = .600, respectively). The mean E2, FSH, and LH levels were elevated in the POR group (p < .001, for all). The mean AFC and AMH concentration were significantly reduced in the POR group (p < .001, for both). In addition, there was a significant increase in the serum alarin level in the POR group (p < .001). Pearson’s analysis revealed that the mean BMI value of the POR group had a weak and negative correlation (r = 0.318, p = .046). Also, there was no correlation between serum alarin with E2 and FSH levels in both study groups. A weak and positive correlation was found between serum alarin and LH concentration only in the POR group (r = 0.318, p = .045). The mean AMH and AFC values were not significantly correlated with serum alarin levels. Conclusion The circulating alarin level was significantly elevated in infertile women with POR patterns. In addition, the alarin level was significantly correlated with the serum LH concentration in the POR pattern.

P0521 - Effects of platelet-rich plasma on spermatogenesis and hormone production in an experimental testicular torsion model

Background Platelet-rich plasma is a biological instrument rich in growth factors and cytokines. Objectives The aim of this study was to investigate the effect of platelet-rich plasma on spermatogenesis and hormone production in an experimental testicular torsion model. Materials and methods The rats were randomly divided into three groups, including six rats in each group as follows: the first group as the sham group; the second group as the ischemia/reperfusion + Saline group and the third group as the ischemia/reperfusion + platelet-rich plasma group. The left testicles of the ischemia/reperfusion + Saline and ischemia/reperfusion + platelet-rich plasma group were kept in four-hour torsion. Then, the left testicles of ischemia/reperfusion + Saline and ischemia/reperfusion + platelet-rich plasma groups were detorsioned, and intra-testicular 1 cc saline (ischemia/reperfusion + Saline) and 1 cc platelet-rich plasma (ischemia/reperfusion + platelet-rich plasma) were injected. At one month, blood samples were taken from all groups for hormonal evaluation and left orchiectomy was performed. Results The mean follicle-stimulating hormone level of ischemia/reperfusion + Saline group was significantly higher than ischemia/reperfusion + platelet-rich plasma group (7.78 ± 0.23 vs 6.18 ± 0.28 nmol/l, respectively, P = .004). The mean LH level of ischemia/reperfusion + platelet-rich plasma group was significantly lower than ischemia/reperfusion + Saline group (3.63 ± 0.28 vs 5.68 ± 0.21 nmol/l, respectively, P = .004). The mean total testosterone level of ischemia/reperfusion + platelet-rich plasma group was significantly higher than ischemia/reperfusion + Saline group (8.05 ± 0.24 vs 5.78 ± 0.23 nmol/l, respectively, P = .004). The mean Johnsen scores of ischemia/reperfusion + platelet-rich plasma group were significantly higher than ischemia/reperfusion + Saline group (5.85 ± 0.58 vs 3.93 ± 0.65, respectively, P = .004). The mean Johnsen score of the sham group was significantly higher than ischemia/reperfusion + platelet-rich plasma and ischemia/reperfusion + Saline groups (P = .003 and P = .003, respectively). Discussion and conclusion The platelet-rich plasma has beneficial effects on spermatogenesis and reproductive hormone production in testicular torsion. It is easily accessible and applicable. In the future, intra-testicular platelet-rich plasma injection may be used in testicular torsion after detorsion. However, further experimental and large-scale prospective clinical studies are needed to establish a definitive conclusion on this topic.

Relationship Between 24-Hour Serum LH and Testosterone Concentrations and Their Interrelationships With Other Pituitary Hormones in Healthy Older Men

Background: With ageing, LH levels rise while T levels decline in men, although this decline in T levels could also be caused by a change in health status, including body composition, inflammation, and comorbidities. Not only levels of LH and T change with age, but levels of other pituitary hormones also change concomitantly with age. It could be hypothesized that these hormonal changes are synchronized with each other.Objective: In this study, we aimed to determine the relationship between 24-h serum LH and T concentrations in healthy older men. Besides, we aimed to determine which health factors, including body composition, metabolic and inflammatory markers, and LH-T related markers are associated with the strength of this LH-T relationship. Furthermore, we explored the interrelationships between LH and T with 24-h serum concentrations of GH, TSH, cortisol, and ACTH.Design: Hormones were measured in serum samples collected every 10 min during 24 h from 20 healthy men, comprising 10 offspring of long-lived families and 10 control subjects, with a mean (SD) age of 65.6 (5.3) years. We performed cross-correlation analyses to assess the relative strength between two 24-h hormone concentration series for all possible time shifts.Results: A mean (95% confidence interval) maximal correlation coefficient of 0.21 (0.10 – 0.31) at lag time 60 min was found between LH and total T concentrations. Results were comparable for calculated free, bioavailable, or secretion rates of T. Men with strong LH-T cross-correlations had, compared to men with no LH-T relationship, lower fat mass (18.5 (14.9 – 19.7) vs. 22.3 (18.4 – 29.4) kg), waist circumference (93.6 (5.7) vs. 103.1 (12.0) cm), hsCRP levels (0.7 (0.4 – 1.3) vs. 1.8 (0.8 – 12.3) mg/L), IL-6 levels (0.8 (0.6 – 1.0) vs. 1.2 (0.9 – 3.0) pg/mL), and 24-h mean LH levels (4.3 (2.0) vs. 6.1 (1.5) U/L), and stronger LH-T feedforward synchrony (1.5 (0.3) vs. 1.9 (0.2)). Furthermore, T was positively cross-correlated with TSH (0.32 (0.21 – 0.43)), cortisol (0.26 (0.19 – 0.33)), and ACTH concentrations (0.26 (0.19 – 0.32)).Conclusions: LH concentrations were followed by T concentrations/secretion with a delay of 60 min in healthy older men, which is in line with literature. Men with a strong LH-T relationship had more favorable body composition, inflammatory markers, 24-h mean LH levels, and LH-T feedforward synchrony. In contrast, chronological age and 24-h mean T levels were not associated with the strength of the LH-T relationship. This observation could indicate that LH and health markers play a bigger role in determining the strength of LH-T cross-correlations than T and chronological age. Furthermore, we observed positive correlations between T and TSH, cortisol, and ACTH concentrations. These exploratory analyses could indicate that T and other hormones are driven by a common regulator or that there is crosstalk between these hormones.

The Assessment of 5 Polymorphisms in the USP9Y Gene in Infertile Patients with Non-obstructive Azoospermia

Background: Male factor infertility that is the cause of about half of the infertility cases, may occur due to azoospermia. Because spermatogenesis defects may lead to non-obstructive azoospermia (NOA), investigating the factors involved in spermatogenesis, including hormones and genes, is one of the most important aspects in understanding the mechanism of NOA in men. Objectives: Male infertility is a complex disorder that affects a large proportion of men, yet many of its causes are unknown. Clarifying its genetic basis may help to identify the causes of infertility and provide effective treatment for patients. Methods: In this case-control study, single nucleotide polymorphisms (SNPs) located in the USP9Y gene were investigated. Accordingly, 100 healthy men and 100 NOA patients were regarded as control and case groups, respectively. Testis tissue samples were taken during the testicular sperm extraction (TESE) procedure, DNA extraction was done, and ARMS PCR was designed and performed. The rs3212292, rs2032597, rs2032604, rs2032598, and rs717268 polymorphisms in the USP9Y gene were analyzed by Tetra-ARMS PCR. Furthermore, the serum levels of sex hormones were assessed by the ELISA technique. Finally, the obtained data were analyzed by SPSS software. Results: According to the obtained results, there was no significant difference in case of genotype frequency of investigated SNPs between the case and control groups (P ≥ 0.05). The mean FSH and LH levels in the control group were relatively lower than the case group (P ≤ 0.05), whereas no significant difference in the mean testosterone level was observed between the two groups (P ≥ 0.05). Conclusions: Polymorphisms of the assessed SNPs showed no effect on the frequency of azoospermia; Thus, polymorphisms did not increase the risk of NOA and also did not have a protective effect against the disease. Also, the results showed that the serum levels of FSH and LH increased in NOA patients.

The Hypothalamo-Pituitary-Gonadal System in Men with Ischemic Heart Disease

Purpose: This Research was to study the functional state of the hypothalamic-pituitary gland-gonadal system in males with Ischemic Heart Disease (IHD), as well as the effect of hormonal correction of the revealed disorders on factors of pathogenesis and the course of IHD.&#x0D; Materials and Methods: We examined 120 males with IHD of different severity of disease, aged 20-50 years and 34 healthy males of the same age with normal body weight and with uncompromised history for diseases of the endocrine system. Among those surveyed were not included alcoholou anabolizant steroids abusers with severe somatic and inflammatory diseases of the genital organs (prostatitis, urethritis, orchitis). The body weight of patients fluctuated within ± 10% of the ideal. At the time of the examination, all the patients were married, 2 were divorced, and all had children.&#x0D; Depending on the age, the examinees were divided into 2 groups: the first - 44 patients in the age of 20-35 years, the second - 76 patients 36-50 years old.&#x0D; The patient was examined in Ashgabat city, Hospital with the Scientific- clinical Center of Cardiology (Turkmenistan).&#x0D; Results: In the I age group, 8 patients had post infarction cardiosclerosis, in which there was a slight decrease in sexual activity, in others - angina pectoris of different functional classes (there was no violation of sexual activity).&#x0D; The average content of Т in the fasting serum of males with IHD of I group was significantly lowered, were noted a statistically unreliable tendency to increase of FSH and a statistically significant increase in the LH/T ratio. The mean levels of LH, PRL and E2 did not differ from the corresponding indices in healthy individuals.&#x0D; Conclusion: Patients with IHD have decreased testosterone secretion, especially at a young age.

Prognosis of prolonged disorders of consciousness outcome based on the determination of certain hormones and natriuretic peptide

Introduction: Changes in hormonal status in patients with prolonged disorders of consciousness (PDC) remain poorly understood. There are no studies devoted to the investigation of prognostic value of hormonal changes to predict the outcome which is primarily due to a relatively small number of patients, different variants of structural brain disorders in vegetative state (VS) patients, concomitant somatic pathology including typical complications.The objective: to study the correlation between outcomes in VS patients with hormonal status and the level of natriuretic peptide. Subjects and methods. 275 patients in VS were examined for the time period from 2007 to 2017. 152 patients had sequela of traumatic brain injury (TBI) and 123 patients suffered from hypoxic brain damage. All patients matched the international criteria of VS diagnosis. In all patients, blood samples were collected during the first week after admission to ICU to test hormones and natriuretic peptide levels. ACTH, cortisol, TSH, free T3 and T4 , STH, prolactin and natriuretic peptide were tested in the period from 2 to 4 months of staying in VS. In men, the level of total testosterone, LH and FSH was additionally tested. The obtained data were compared with VS outcome.Results. The tested hormones were stably insufficient only in few VS patients. None of the tested hormones of the hypothalamic-pituitary-adrenal axis made a reliable criterion for predicting VS outcome. The tendency of disrupted rhythm of cortisol secretion was found to be most frequent and consistent, with higher rates in the evening hours. The average value of STH was higher in men with sequela of traumatic brain injury who had recovered consciousness versus those who remained in VS. A significant decrease in testosterone levels, regardless of age, was found in the patients with TBI sequela. Mean levels of LH were higher in patients with TBI sequela and hypoxia who remained unconscious versus patients who later restored consciousness. The average level of FSH was higher in patients who had recovered consciousness compared to those who remained in chronic VS. The increased level of natriuretic peptide was observed both in patients who remained in chronic VS and in those who restored consciousness.Conclusions. When investigating levels of certain hormones, no specific endocrine background characterizing this category of patients was found. Abnormal rhythms of some hormones secretion, in particular cortisol, can be considered typical of VS patients especially patients with TBI sequela.

Voice break in boys-temporal relations with other pubertal milestones and likely causal effects of BMI.

STUDY QUESTIONHow is timing of voice break related to other male pubertal milestones as well as to BMI?SUMMARY ANSWERWe provide a comprehensive temporal analysis of male pubertal milestones, including reproductive hormone dynamics, confirm voice break as a late milestone of male puberty and report a likely causal relationship between higher BMI and earlier age at voice break in men.WHAT IS KNOWN ALREADYVoice break represents a late pubertal milestone and recalled age at voice break is frequently used in epidemiological studies as a measure of puberty. In contrast, clinical studies use mainly testicular enlargement and/or genital tanner stage as the marker of pubertal onset. However, neither correlation of pubertal milestones nor reproductive hormone dynamics have been assessed in detail previously. Further, although BMI and puberty timing are known to be closely linked, cause and effect between these traits are not known.STUDY DESIGN, SIZE, DURATIONThe study included a population-based mixed cross-sectional and longitudinal cohort (2006–2014, COPENHAGEN Puberty Study) of 730 healthy Danish boys. Data for 55 871 male research participants from the 23andMe study were obtained, including genome-wide single nucleotide polymorphism data and age at voice break.PARTICIPANTS/MATERIALS, SETTING, METHODSWe performed a detailed evaluation of pubertal milestones and reproductive hormone levels (study population 1). A Mendelian randomization (MR) approach was used to determine the likely causal link between BMI and timing of voice break (study population 2).MAIN RESULTS AND THE ROLE OF CHANCEVoice break occurred at mean age 13.6 (95% CI: 13.5–13.8) years. At voice break, mean (95% CI) testosterone levels, LH levels and bi-testicular volume were 10.9 (10.0–11.7) nmol/L, 2.4 (2.2–2.5) IU/L and 24 (23–25) mL, respectively. Voice break correlated moderately strongly with timing of male pubertal milestones, including testicular enlargement, gonadarche, pubarche, sweat odor, axillary hair growth and testosterone above limit of detection (r2 range: 0.43–0.61). Timing of all milestones was negatively associated with age-specific BMI (all P ≤ 0.001). MR analyses inferred likely causal effects of higher BMI on earlier voice break in males (−0.35 years/approximate SD, P < 0.001).LIMITATIONS, REASONS FOR CAUTIONParticipation rate of the population-based cohort was 25%. Further, boys that were followed longitudinally were examined approximately every 6 months limiting the time resolution of pubertal milestones. Using adult BMI as exposure instead of prepubertal BMI in the MR analysis and the known inaccuracies of the testosterone immunoassay at low testosterone levels may be further limitations.WIDER IMPLICATIONS OF THE FINDINGSWe provide valuable normative data on the temporal relation of male pubertal milestones. Further, the likely causal relationship between BMI and puberty timing highlights the importance of preventing obesity in childhood.STUDY FUNDING/COMPETING INTEREST(S)This work was supported by Danish Agency for Science, Technology and Innovation (09-067 180); Danish Ministry of the Environment, CeHoS (MST-621-00 065); Capital Region of Denmark (R129-A3966); Ministry of Higher Education and Science (DFF-1331-00 113); Innovation Fund Denmark (InnovationsFonden, 14-2013-4); The International Center for Research and Research Training in Endocrine Disrupting Effects of Male Reproduction and Child Health. B.H., F.R.D., J.R.B.P. and K.K.O. are supported by the Medical Research Council (MC_UU_12015/2). The 23andMe study is supported by the National Human Genome Research Institute of the National Institutes of Health (R44HG006981). Members of the 23andMe Research Team are employees of 23andMe, Inc. and hold stock or stock options in 23andMe.TRIAL REGISTRATION NUMBERNCT01411527