Mean FSH Research Articles

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.

Study the effect of piroxicam treatment for intrauterine insemination using density gradient for sperm preparation

Back ground: Endometrium contraction following intra-uterine insemination(IUI) was considered one of implantation failure causes.Aim of the study: The primary purpose of the study was to ascertain the effect of piroxicam treatment on pregnancy outcome following six days of intrauterine insemination. Materials and Methods: The study was accomplished in the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University through the period from Nov. 2020 till Oct. 2021 .Forty infertile couples were selected randomly for IUI and divided into 20 couple treated with piroxicam and other 20 couples were not treated with piroxicam. The semen samples were prepared by discontinuous density gradient. Results: There was no significant differences regarding mean FSH level (p=0.769), LH level(p=0.79) prolactin level (p=0.299), E2 level (p=0.348) and TSH level (p=0.950)between the studied groups. There was no significant difference in dominant follicles counts, follicular sizes or endothelial thickness between the two groups. The results showed a significant improvement in sperm motility and morphology (p < 0.05) after in vitro activation by density gradient technique compared to before activation. In the piroxicam group ,4 out of 20 women became pregnant while in non-piroxicam group ,3 spouses were pregnant 15%when density gradient activation technique was used. 

P-634 Safety of ART cycles with extremely high estradiol levels - A retrospective cohort study

Abstract Study question Are IVF treatments with extremely high peak estradiol levels during ovarian hyperstimulation associated with higher complication rates? Summary answer Extremely high estradiol levels were not associated with higher major complication rates, but patients were more likely to seek medical care following retrieval. What is known already The freeze-all policy and the increasing popularity of social oocyte freezing, have led to an increase in gonadotropin doses and number of oocytes retrieved and, as a direct effect, higher estradiol levels. Studies reporting major ART complications (OHSS, bleeding, infection, torsion, thromboembolic events) are based on heterogeneous patient populations without stratification by oocyte number or estradiol levels, except for a single study reporting an association between peak estradiol levels and the risk of ovarian torsion. Extreme estradiol levels, are associated with larger ovaries, repeated ovarian punctures, and possibly a thrombogenic effect. Therefore, higher complication rates can be expected. Study design, size, duration A retrospective cohort study including patients from two large medical centers treated between 2019-2021. Participants/materials, setting, methods A study group (239 patients) with extremely high peak estradiol levels (>20,000 pmol/L on the day of ovulation induction, or > 15,000 pmol/L on the previous day) and a control group (208 patients) with normal range estradiol levels (3000-12000pmol/L), treated at two large units were included. Patients were surveyed about complaints and medical care related to ovum pick up (OPU), and medical files were reviewed. Complication rates and the need for medical assistance were compared. Main results and the role of chance Several differences between the study and control group were observed as a consequence of the study design: Mean age was 33.01±5.14 vs. 34.57±4.52 (p = 0.01), Mean peak estradiol levels was 26645.34±8592.56 vs.7229.750±2329.20 (p < 0.001), mean number of oocytes were 27.55±13.46 vs. 11.63±5.77 (p < 0.001) for the study and control group respectivly. More patients in the control group underwent fertility preservation. Mean FSH levels and total gonadotropin dosage were higher in the control group. Major complications [3 (1.25%) in the study group vs. 1 (0.96%) in the control group] were similar between groups (p = 0.62). In the study group Two patients were diagnosed with ovarian torsion (0.83%), and one (0.41%) with early severe OHSS as a result of a positive BhCG related to an undiagnosed pregnancy prior to OPU. All three were admitted (1.25%). In the control group one patient was diagnosed with PID requiring inpatient IV antibiotic treatment (0.48%). One more patient was admitted with severe abdominal pain following OPU, for a total of 2 admissions (0.96%) (p = 1). 33 patients (13.8%) in the study group and 10 (4.8%) in the control group sought medical care after OPU, mostly due to abdominal pain, without further workup or hospitalization (p = 0.001). Limitations, reasons for caution A retrospective study with possible recall bias. Major adverse events are rare in IVF and may not be fully captured in the study population. Wider implications of the findings Based on our results, extremely high estradiol levels during ovarian hyperstimulation were not associated with thromboembolic events, higher major complication or hospitalization rates, thus may be considered safe. Nevertheless, patients may be informed of possible higher rates of discomfort, mostly abdominal pain. Larger studies are warranted to confirm our results. Trial registration number 0090-21-ASF

May endocan be a new biomarker in the diagnosis of endometriosis?

AimAs known, inflammatory substances are considered to have a role in the onset and progression of endometriosis. In this study, we aimed to find a biomarker that can be used in the diagnosis of endometriosis by investigating the serum levels of endocan, which is a substance that we know to have an important role in angiogenesis and inflammation, in patients with endometriosis. Study Design45 patients between the ages of 18-40 with the diagnosis of stage 3-4 endometriosis and whose postoperative histopathological tissue diagnoses were endometriosis were included in the study as study group. As the control group, a total of 45 healthy women between the ages of 18-40 were included in the study. The two groups were statistically compared. ResultsThere was no statistically significant difference between the two groups in terms of age, BMI, LH, E2, and mean Hb values. It was observed in the examination of the endocan levels that the mean values in the study (endometriosis, patient group) group were statistically and significantly higher compared to the control (healthy) group (p:0.000). Also, mean FSH and Ca125 levels were determined to be statistically and significantly higher in the endometriosis group (p:0.042 and p:0.000). ConclusionIn this study, we found a statistically significant correlation between the levels of serum endocan and endometriosis. As the results, endocan can be used as a new biomarker to diagnose patients with endometriosis or in their follow up period. Much more comprehensive future studies are needed on this subject.

Acute progesterone feedback on gonadotropin secretion is not demonstrably altered in estradiol-pretreated women with polycystic ovary syndrome.

Women with polycystic ovary syndrome (PCOS) demonstrate gonadotropin‐releasing hormone (GnRH) pulse generator resistance to suppression with 7 days of progesterone and estradiol administration. It remains unknown whether such women demonstrate impairments in acute progesterone negative feedback on LH pulse frequency or progesterone positive feedback on gonadotropin release. This was a randomized, double‐blind, placebo‐controlled crossover study designed to test the hypothesis that acute, progesterone‐related suppression of LH pulse frequency and progesterone‐related augmentation of gonadotropin release are impaired in PCOS. Twelve normally cycling women and 12 women with PCOS completed study. Volunteers were pretreated with transdermal estradiol (0.2 mg/day) for 3 days and then underwent a frequent blood sampling study (20:00–20:00 h), during which they received micronized progesterone (100 mg) or placebo at 06:00 h. In a second study admission, volunteers received the intervention they did not receive during the first admission, but the protocol was otherwise identical. The primary outcome measures were LH secretory characteristics and circulating gonadotropin concentrations. Exogenous progesterone did not reduce LH pulse frequency in either group. Mean LH, pulsatile LH secretion, LH pulse mass, and mean FSH increased more with progesterone compared to placebo in both groups. Although trends toward less pronounced changes in LH pulse mass and pulsatile LH secretion were observed in the PCOS group, these differences were not statistically significant. In summary, exogenous progesterone did not suppress LH pulse frequency within 12 hours in estradiol‐pretreated women, and the positive feedback effect of progesterone on gonadotropin release was not demonstrably impaired in PCOS.New & NoteworthyThis study indicated that exogenous progesterone does not reduce LH pulse frequency within 12 h in women with PCOS, but progesterone acutely increased gonadotropin in these women. This study suggested that progesterone‐related augmentation of gonadotropin release may be impaired in PCOS compared to normally cycling women, but this finding was not statistically significant.

The Combination of High Calorie Diet and Moderate Interval Training Decreases Follicle Stimulating Hormone (FSH) Level In Rats

Background: Obesity can cause leptin and insulin resistance which will affect the production of GnRH and FSH. One of risks factor of obesity is high calorie intake chronicaly. Exercise plays a role in the regulation of blood calorie and body weight. However, the effect of exercise on subjects with a high-calorie diet on FSH is unknown. This study aims to determine the effect of a high calorie diet with moderate intensity interval training (MIIT) on FSH level in female rats. Method: Sample was 27 female rats (Rattus norvegicus) wistar strain were divided into 3 groups. The negative control group (standard diet), the positive control group (high calorie diet), and the MIIT group (high calorie diet and moderate-intensity exercise with intervals). Measurement of FSH using the ELISA method after treatment for 4 weeks. Result: Statistical analysis showed that there was a significant difference in FSH level with a value (p = 0.001). The mean FSH was 25.32 mIU/ml ? 7.89 in the negative control group, 19.71 mIU/ml ? 6.55 in the positive control group, and 12.12 mIU/ml ? 3.64 in the MIIT group. The decrease FSH in the positive control and MIIT group due to a high calorie diet caused insulin and leptin resistance which affected GnRH. Conclusion: Moderate interval training decreases the FSH level may due to the level of physical stress.

FSH levels and testicular volumes are associated with the severity of testicular histopathology in men with non-obstructive azoospermia

PurposeThe purpose of this study is to assess a potential association between FSH levels and testicular volumes with the severity of testicular histopathology on testicular biopsy in men with non-obstructive azoospermia (NOA) undergoing microdissection testicular sperm extraction (microTESE).MethodsA retrospective chart review was performed from the electronic health records of men who underwent microTESE with NOA.ResultsEighty-six men with NOA underwent microTESE with concomitant testicular biopsy for permanent section to assess the testicular cellular architecture. The histopathological patterns were categorized by severity indicating the odds of sperm retrieval into 2 categories. The unfavorable category included Sertoli cell only pattern and early maturation arrest (n = 50) and the favorable category included late maturation arrest and hypospermatogenesis patterns (n = 36). In the men with unfavorable histopathologic patterns, the mean FSH level was 22.9 ± 16.6 IU/L, and the mean testicular volume was 10.4 ± 6.0 cc. This was in comparison to men with favorable histopathologic patterns revealing a mean FSH level of FSH 13.3 ± 12.0 with a mean testicular volume of 13.3 ± 5.9 cc. There was a statistically significant higher FSH level in men with unfavorable histopathology than favorable (p = 0.004) as well as a significant smaller mean testicular volume in men with unfavorable histopathology (p = 0.029).ConclusionsHigher serum FSH levels and smaller testicular volumes are associated with more severe testicular histopathological patterns in men with NOA.

PD15-07 DOES AGE OF TESTIS LOSS IMPACT COMPENSATORY HYPERTROPHY OF THE REMAINING TESTIS? A STUDY OF NEONATAL, PREPUBERTAL AND ADULT MICE

You have accessJournal of UrologyPediatric Urology I (PD15)1 Sep 2021PD15-07 DOES AGE OF TESTIS LOSS IMPACT COMPENSATORY HYPERTROPHY OF THE REMAINING TESTIS? A STUDY OF NEONATAL, PREPUBERTAL AND ADULT MICE Daniel E. Nassau, Eliyahu Kresch, Shathiyah Kulandavelu, Oleksandr N. Kryvenko, Miguel A. Castellan, Himanshu Arora, and Ranjith Ramasamy Daniel E. Nassau Daniel E. Nassau More articles by this author , Eliyahu KreschEliyahu Kresch More articles by this author , Shathiyah KulandaveluShathiyah Kulandavelu More articles by this author , Oleksandr N. KryvenkoOleksandr N. Kryvenko More articles by this author , Miguel A. CastellanMiguel A. Castellan More articles by this author , Himanshu AroraHimanshu Arora More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001997.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The long-term sequelae of partial testicular loss on male reproductive function is unknown. Compensatory testicular hypertrophy occurs when testis loss is before puberty; however, it is unclear if the degree of growth differs by prepubertal age. Using a mice model, we hypothesized that contralateral testicular compensatory growth would be greater if unilateral orchiectomy was done at younger prepubertal age and lesser or absent if done post-puberty. METHODS: Hemicastration was performed in C57BL/6 mice at a neonatal, prepubertal or postpubertal period between days of life (DOL) 2-4, 20-22 and 42-44, respectively. These mice and a control group whom did not undergo a procedure were sacrificed after physical maturity (DOL 80) and remaining testis was removed. We evaluated FSH levels, histology, body weight (BW), testis weight (TW) and testis long-axis (calipers). The larger testis was recorded for controls. ANOVA was used to determine statistical significance (p<0.05). RESULTS: Median neonatal and prepubertal TW were significantly greater than control; however, only the median TW/BW ratio in the neonatal group was significantly higher than control, implying the greatest degree of compensatory growth happened with earlier testis loss. Neonatal TW was also significantly greater than postpubertal TW. No difference in BW or testis long axis length in any group was detected (Figure 1). H&E evaluation showed similar degree of spermatogenesis and Leydig cell concentration among all mice. Mean FSH (ng/mL) was highest in postpubertal (105.9) followed by prepubertal (98.5), neonate (93.9) and control (61.2), suggesting lower global sperm production in mice with testis loss at a later age. CONCLUSIONS: Contralateral testicular hypertrophy occurred if testis loss was during the prepubertal period and our data implies that earlier testis loss may have a greater degree of compensation and maintained reproductive function. This challenges the notion that the testis remains quiescent during childhood as there must be a signaling cascade to promote testicular hypertrophy. To our knowledge, this is the first study to successfully perform hemicastration in neonatal mice. This model will be used to study molecular mechanisms that influence testicular growth after unilateral testis loss . Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e274-e275 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Daniel E. Nassau More articles by this author Eliyahu Kresch More articles by this author Shathiyah Kulandavelu More articles by this author Oleksandr N. Kryvenko More articles by this author Miguel A. Castellan More articles by this author Himanshu Arora More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement Loading ...

P–587 Highly purified human menopausal gonadotrophin (HP-hMG, Menopur) as a ready-to-use solution for injection in pre-filled pen is bioequivalent to HP-hMG powder for reconstitution

Abstract Study question Are serum FSH levels after single subcutaneous dosing of HP-hMG in a liquid formulation and a powder formulation bioequivalent? Summary answer The 90% CIs for the geometric mean ratios of serum FSH AUCt and Cmax were both within 0.8000–1.2500, thus the two formulations are bioequivalent. What is known already For several decades, HP-hMG (Menopur) has been used for the treatment of infertility; its efficacy and safety compared to other gonadotropins have been consistently demonstrated in several prospective, randomised controlled trials and meta-analyses (Deeks et al 2018; Bordewijk et al 2019). Menopur powder for reconstitution is available in multidose and single dose formulations. Up to 3 single dose vials (each containing 75 IU) may be dissolved into 1 mL solvent for administration. Recently, and for the first time, Menopur has been successfully formulated in a stable, ready-to-use solution for injection, which may be administered by a pre-filled pen. Study design, size, duration This was a randomised, two-way crossover, single dose, bioequivalence trial comparing Menopur liquid injected by pre-filled pen, with Menopur powder injected by conventional syringe and needle. The primary endpoints were AUCt and Cmax of baseline-adjusted FSH. Pituitary-suppressed, healthy women were randomised to receive one treatment sequence including a single subcutaneous injection of 450 IU Menopur liquid (600 IU/0.96 mL) and of 450 IU Menopur powder by two subcutaneous injections of 225 IU in 1 mL. Participants/materials, setting, methods Blood samples were collected pre- and post-dose, until 9 days after each injection. The PK parameters of FSH and hCG were assessed by noncompartmental methods with adjustment for endogenous pre-dose levels. Highly sensitive and specific electrochemiluminescence immunoassays were used for quantification and the LLOQ of the FSH and hCG assays were 1.47 mIU/mL and 0.5 mIU/mL, respectively, and the total validated CV was within 5% for both assays. Main results and the role of chance In total, 76 women were randomised and 56 completed the trial. The main reason for discontinuation was insufficient pituitary suppression prior to the second administration of HP-hMG. The mean FSH and hCG serum concentration-time profiles were comparable between the two HP-hMG formulations. The geometric mean ratios and 90% confidence intervals of FSH for HP-hMG liquid versus HP-hMG powder were 1.12 [1.0562; 1.1889] for AUCt and 1.17 [1.0946; 1.2490] for Cmax, showing that the two formulations were bioequivalent. Maximal serum FSH concentrations were reached at 18.19 h for HP-HMG liquid and 15.55 h for HP-hMG powder. In addition to FSH, the PK parameters for hCG were compared between the two HP-hMG formulations. The geometric mean ratios and 90% confidence intervals for HP-hMG liquid versus HP-hMG powder were 0.93 [0.86; 1.01] for AUCt and 0.94 [0.86; 1.02] for Cmax. There was no difference between the two groups in the incidence or severity of adverse events, and both preparations were well tolerated. Mild injection site reactions were less common after administration of HP-hMG liquid by a single injection compared to HP-hMG powder by two injections and were mostly related to pain and erythema after drug administration. Limitations, reasons for caution This bioequivalence study is based on the comparison of single dose administrations in healthy female volunteers of reproductive age. Wider implications of the findings: The new HP-hMG solution for injection in a pre-filled pen will deliver the efficacy and safety of Menopur in a convenient delivery device. Trial registration number NA

P–796 Trial of Autologous Marrow derived Stem Cell Ovarian Transplantation (TAMSCOT) in young infertile women with diminished ovarian reserve for ovarian rejuvenation – HOPE still persists

Abstract Study question Does autologous bone marrow derived stem cell (BMDSC) ovarian transplantation optimize ovarian reserve parameters in young infertile women with diminished ovarian reserve (DOR) ? Summary answer The autologous stem cell ovarian transplantation (ASCOT) improves AFC and AMH by facilitating the recruitment of existing dormant follicles in young women with DOR. What is known already Oocyte donation is the practical therapeutic option when patients with premature ovarian ageing desire pregnancy. It involves significant psychological burden in terms of not able to have their own biological child. ASCOT has opened new doors in poor responders and premature ovarian insufficiency through its beneficial effects on ovarian reserve and IVF outcomes. However recent studies have shown contradictory results in terms of its efficacy. No prior study has been contemplated in DOR group Study design, size, duration An open label non randomized controlled trial was conducted at Division of Reproductive Medicine in collaboration with stem cell facility at tertiary care institute. Forty two infertile women less than 35 years age with DOR (AFC&amp;lt;5, AMH&amp;lt;1.2ng/ml and /or high FSH&amp;gt;8IU/l) were enrolled in the study during a period from January 2020 to December 2020. 20 women who did not opt for the intervention were treated as control group whereas 22 women received the intervention. Participants/materials, setting, methods Baseline hormonal profile ( Day 2 FSH, estradiol, AMH and AFC) was done in all patients. Women with abnormal uterine cavity, endometriosis, prior ovarian surgery, abnormal karyotype were excluded. Bone marrow aspiration followed by mesenchymal stem cells isolation was performed. The stem cells were transplanted in both the ovaries through transvaginal route on the same day. Follow up visits were planned at one and six months to assess ovarian reserve parameters. Main results and the role of chance The mean age, BMI and duration of infertility were comparable between the control and study group (29.5±3.34vs29.36±2.95years, 21.51±1.40vs21.87±1.93kg/m2, 6.9±1.94vs7. 04±3.67 years) . The positive response in terms of improved AMH and AFC was seen in 68% (15/22) patients. The mean number of stem cells injected in these women were 77.71±25.33 million. At first follow up, there was no significant difference between mean FSH, estradiol levels and mean right and left ovarian volume (9.23±3.95 vs 9.02±3.92mIU/l, 61.46±29.25 vs 68.12±62.52 pg/ml, 2.82±2.18 vs 2.44±1.25 cc, 2.02±1.54 vs 2.72±1.06 cc, p &amp;lt; 0.05). There was significant increase in AMH and AFC values as compared to baseline (0.79±0.43 vs 1.26±0.82ng/ml, p = 0.03; 3.47±1.30 vs 6.40±2.23, p &amp;lt; 0.001). At second follow up visit, the significant increase in ovarian reserve persisted for AMH and AFC (0.79±0.43 vs 1.22±0.76 ng/ml, p = 0.02; 3.47±1.30 vs 6.93±1.71,p&amp;lt;0.001). There was no significant difference between serum FSH , Estradiol and ovarian volume. None of the patients developed any complication and the improvement in AFC and AMH persisted during 10 month follow up period. Limitations, reasons for caution The limitation of present study is small sample size and non randomization. However, time period for which positive effect lasts has not been documented in earlier studies. This study is currently being endeavored, and women with improved ovarian reserve are followed up for any spontaneous conception or following assisted reproduction. Wider implications of the findings: The present study demonstrates beneficial role of stem cells in improving ovarian reserve parameters in women with DOR with no acquired cause. If supported by future randomized clinical studies, it could represent a paradigm shift for fertility treatment in these women providing an opportunity to have their own biological child. Trial registration number CTRI/2020/01/022726

P-796 Trial of Autologous Marrow derived Stem Cell Ovarian Transplantation (TAMSCOT) in young infertile women with diminished ovarian reserve for ovarian rejuvenation – HOPE still persists

Abstract Study question Does autologous bone marrow derived stem cell (BMDSC) ovarian transplantation optimize ovarian reserve parameters in young infertile women with diminished ovarian reserve (DOR) ? Summary answer The autologous stem cell ovarian transplantation (ASCOT) improves AFC and AMH by facilitating the recruitment of existing dormant follicles in young women with DOR. What is known already Oocyte donation is the practical therapeutic option when patients with premature ovarian ageing desire pregnancy. It involves significant psychological burden in terms of not able to have their own biological child. ASCOT has opened new doors in poor responders and premature ovarian insufficiency through its beneficial effects on ovarian reserve and IVF outcomes. However recent studies have shown contradictory results in terms of its efficacy. No prior study has been contemplated in DOR group Study design, size, duration An open label non randomized controlled trial was conducted at Division of Reproductive Medicine in collaboration with stem cell facility at tertiary care institute. Forty two infertile women less than 35 years age with DOR (AFC&amp;lt;5, AMH&amp;lt;1.2ng/ml and /or high FSH&amp;gt;8IU/l) were enrolled in the study during a period from January 2020 to December 2020. 20 women who did not opt for the intervention were treated as control group whereas 22 women received the intervention. Participants/materials, setting, methods Baseline hormonal profile ( Day 2 FSH, estradiol, AMH and AFC) was done in all patients. Women with abnormal uterine cavity, endometriosis, prior ovarian surgery, abnormal karyotype were excluded. Bone marrow aspiration followed by mesenchymal stem cells isolation was performed. The stem cells were transplanted in both the ovaries through transvaginal route on the same day. Follow up visits were planned at one and six months to assess ovarian reserve parameters. Main results and the role of chance The mean age, BMI and duration of infertility were comparable between the control and study group (29.5±3.34vs 29.36±2.95years, 21.51±1.40vs21.87±1.93kg/m2, 6.9±1.94vs7.04±3.67 years). The positive response in terms of improved AMH and AFC was seen in 68% (15/22) patients. The mean number of stem cells injected in these women were 77.71±25.33 million. At first follow up, there was no significant difference between mean FSH, estradiol levels and mean right and left ovarian volume (9.23±3.95 vs 9.02±3.92mIU/l, 61.46±29.25 vs 68.12±62.52 pg/ml, 2.82±2.18 vs 2.44±1.25 cc, 2.02±1.54 vs 2.72±1.06 cc, p &amp;lt; 0.05). There was significant increase in AMH and AFC values as compared to baseline (0.79±0.43 vs 1.26±0.82ng/ml, p = 0.03; 3.47±1.30 vs 6.40±2.23, p &amp;lt; 0.001). At second follow up visit, the significant increase in ovarian reserve persisted for AMH and AFC (0.79±0.43 vs 1.22±0.76 ng/ml, p = 0.02; 3.47±1.30 vs 6.93±1.71,p&amp;lt;0.001). There was no significant difference between serum FSH, Estradiol and ovarian volume. None of the patients developed any complication and the improvement in AFC and AMH persisted during 10 month follow up period. Limitations, reasons for caution The limitation of present study is small sample size and non randomization. However, time period for which positive effect lasts has not been documented in earlier studies. This study is currently being endeavored, and women with improved ovarian reserve are followed up for any spontaneous conception or following assisted reproduction. Wider implications of the findings The present study demonstrates beneficial role of stem cells in improving ovarian reserve parameters in women with DOR with no acquired cause. If supported by future randomized clinical studies, it could represent a paradigm shift for fertility treatment in these women providing an opportunity to have their own biological child Trial registration number CTRI/2020/01/022726

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.

INTERRELATIONSHIP OF AMH WITH CLINICAL, HORMONAL AND SONOGRAPHIC PARAMETERS IN PATIENTS WITH POLYCYSTIC OVARIAN DISEASE

INTRODUCTION: Polycystic ovary syndrome (PCOS) is a frequently encountered problem in reproductive endocrinology, affecting approximately 6% of women of reproductive age. anti-mullerian hormone (AMH) also known as Mullerian inhibiting, substance is produced by granulosa cells of the preantral and small antral ovarian follicles in women and reect the ovarian reserve. Women with PCOS will often have a high number of antral follicles and, as a result, an equally high level of AMH in their blood. it has an inhibitory inuence on the actions of FSH and positively correlated with LH. Various studies demonstrated that oligo/anovulatory women with PCOS have signicantly higher serum concentrations of AMH. MATERIAL AND METHOD: This is a retro respective study of 54 PCOS patients from rst January 2019 to 13 April 2021 at a private gynaecology clinic in district Shivpuri. Data were collected from the medical records of the patients including age, height, weight, waist circumference, BP and modied Ferriman gallway score for hirsutism, Biochemical and hormonal values like LH, FSH, LH/FSH ratio, TSH, Prolactin and AMH, and lipid prole values were also obtained from the records. Abdominal or Vaginal ultrasound was used to assess the ovarian volume antral follicular count. RESULTS: In our study, the age of patients ranges from 20-39 years and a majority of the patients were in the age group of 21-30 years 75.28%) The Mean age of PCOS patients was 23.89. In our study, the mean cycle length of patients was 54.08 and it ranged from 28-190 days. The mean 2 BMI 25.8kg/m2 and modied FG score were 8.1. On ultrasound mean antral follicle count was 26.2 and the mean ovarian volume was 9.4 cm . In hormonal studies mean LH was 11.5, mean FSH was 3.4 and AMH was 4.34 and Testosterone was 2.1. In bio-chemical parameters increased Cholesterol, triglycerides, LDL, VLDL level and decreased HDL level were found in 31.48% of patients and with increased AMH. CONCLUSION: In our study, we found increased AMH concentration in correlation with increased cycle length in cases of oligomenorrhea, clinical hyperandrogenism, increased LH, increased AFC and increased ovarian volume. Our study concluded that AMH levels as an adjunct to existing Rotterdam criteria for diagnosis of PCOS had good diagnostic potential.

CLINICAL, METABOLIC, ENDOCRINE AND SONOGRAPHIC CHARACTERISTICS OF OBESE ADOLESCENT WITH PCOS

Introduction- PCOS is a complex metabolic, endocrinopathy and reproductive disorder that results in the production of androgens and is associated with insulin resistance. Adolescents with PCOS are more obese than normal adolescents and have an increased risk of metabolic syndrome. In obese adolescent increasing abdominal adiposity, worsening insulin sensitivity and Dyslipoproteinemiadyslipoproteinemia give rise to NAFLD in adulthood. This study aimed to investigate the association between PCOS, obesity, NAFLD and Metabolic syndrome in adolescents with PCOS. MATERIALAND METHOD- Retrospective study of 24 obese adolescent PCOS patients. with mean BMI 32.89 kg/m2. Data were taken from 1st January 2015 to 31st December 2020. PCOS patients were diagnosed according to Rotterdam 2003 criteria. Data were collected from the medical records of the patients including clinical history, height, weight, blood pressure, waist circumference and Modied Ferriman-Gallway score for hirsutism. Laboratory values were obtained ultrasound nding of polycystic ovarian disease and fatty liver were obtained from the records. RESULTS:- 2 In our study, the mean age was 15.2 yr The mean body mass index was 32.89 kg/m and the mean age of menarche was 12.3 yr. Hirsutism was present in 75% by modied FG Score system, Family history of PCOS was present in 29.16%, Family history of Cardiovascular disease was present in 12.5%. Afamily history of diabetes mellitus was present in 20.83%. The majority 87.5% of the obese adolescent girl's were presented with menstrual problems. mostly 70.83% with a history of Oligomenorrhea. Waist circumference, &gt;0.80 was present in 87.5%, 50% of patients were hypertensive. 20.83% were prediabetes, 54.16% of patients were found to have dyslipidaemia and 45.83% of patients had Metabolic syndrome. In our mean LH was 10.6 IU/L, mean FSH was 5.8 IU/L, mean Serum testosterone 18.5 ng/ml, mean Serum prolactin was 21.2 ng/mL and mean Serum TSH was 3.3 µIU/mL.75 % of patients had Polycystic appearing ovarian morphology in USG.45.83 % of obese PCOS patient had NAFLD in USG and 33.33% of obese patients had abnormal liver enzymes with NAFLD in USG nding. CONCLUSION:-In our study, we found both metabolic syndrome and NAFLD were frequent in patients with obese adolescent PCOS conrming a relevant clinical association between these three conditions. This study highlights the importance of preventing obesity during the management of adolescent PCOS. The importance of obesity counselling in obese adolescent women is a must to reduce the risks associated with metabolic syndrome. The therapeutic intervention combined with lifestyle modication may provide better treatment for adolescent PCOS.

Comparison of Clinical, Metabolic and Hormonal Effects of Metformin Versus Combined Therapy of Metformin With Myoinositol Plus D-Chiro-Inositol in Women With Polycystic Ovary Syndrome (PCOS): A Randomized Controlled Trial.

ObjectiveTo compare the effects of metformin alone versus combined therapy of metformin with myoinositol (MI) plus D-chiro-inositol (DCI) in women with polycystic ovary syndrome (PCOS).Materials and methodsThis is a prospective, non-blinded randomized controlled trial conducted in newly diagnosed PCOS women aged 18 to 45 years. Group I received metformin 500 mg twice a day orally for 6 months while group II received metformin 500 mg twice a day orally along with MI 550 mg plus DCI 150 mg twice daily orally for six months. The primary outcome was a change in clinical, metabolic and hormonal parameters of the two groups from baseline to the end of six months of treatment.ResultsA total of 72 patients were randomized into two groups of 36 patients each. Statistically, a significant difference was seen in terms of mean global acne score (p=0.004) and cycle regularity (p=0.034) after six months of treatment in group II. A significant difference in values of luteinizing hormone (LH) (p=0.002), luteinizing hormone/follicle-stimulating hormone (LH/FSH) ratio (p=0.007), mean cholesterol (p=0.040), mean high-density lipoprotein (HDL) (p=0.049), mean low-density lipoprotein (LDL) (p=0.0001) and postprandial insulin (p=0.005) was also seen in group II at the end of treatment duration. No significant difference was seen between the two groups in terms of mean FSH, mean testosterone, mean dehydroepiandrosterone sulfate (DHEAS), mean triglyceride, mean fasting and postprandial blood sugar, fasting insulin and homeostatic model assessment of insulin resistance (HOMA-IR) index.ConclusionCombined therapy with metformin and MI plus DCI in women with PCOS and insulin resistance seems promising with the need for further studies with a greater sample size to evaluate the efficacy of this treatment.

Gonadotropin Response to Insulin and Lipid Infusion Reproduces the Reprometabolic Syndrome of Obesity in Lean Women

Introduction: The reprometabolic syndrome of obesity is associated with reduced gonadotropins and impaired LH and FSH response to gonadotropin releasing hormone (GnRH). We sought to reproductive the reprometabolic syndrome in normal weight, eumenorrheic women by infusing a combination of insulin and lipid. Materials and Methods: 15 women, mean age 32 (IQR 26,36) and BMI 21.9 (20.2, 22.9) were recruited with intent to perform early follicular phase, 6-hour infusions of insulin (20-40mg/mU/m2/min) and lipid (Intralipid) or saline infusion (controls); 12 women completed both intended studies and an additional 3 women completed only one of the two studies. The first 4 hours of each study assessed endogenous gonadotropins; at 4hrs, a 75 ng/kg GnRH bolus was administered and sampling continued until 6hrs. Linear mixed model analysis was used to determine differences between Intralipid versus saline on endogenous LH pulse amplitude (primary outcomes), mean FSH, and area under the curve (AUC) response to GnRH (secondary outcomes). Results: LH pulse amplitude, mean FSH, and both AUC responses to GnRH were all reduced by Intralipid/insulin; mean FSH (P=0.03) and AUC for LH (P=0.05) were at or near statistical significance. LH pulse amplitude and response to GnRH were significantly reduced (P=0.04 and 0.02) when one participant with very high LH and AMH levels was excluded. Discussion: Acute infusion of insulin/lipid to eumenorrheic, normal weight women recapitulated the reprometabolic syndrome of obesity. These findings imply that specific circulating factors in obese women contribute to their sub fertility and thus may be amenable to discovery and treatment.

Progesterone Positive Feedback on Pulsatile LH Secretion and FSH Release May Be Blunted in Estradiol-Pretreated Women With PCOS

In women pretreated with estradiol (E2), exogenous progesterone (P4) acutely augments LH and FSH release (P4 positive feedback). Women with PCOS exhibit impaired P4 negative feedback on LH pulse frequency, but it remains unclear whether such women exhibit impaired P4 positive feedback on LH/FSH release. We sought to explore the latter notion as an a priori secondary hypothesis in a study primarily designed to assess whether P4 acutely suppresses LH pulse frequency. We studied 12 women with PCOS and 12 normally-cycling, non-hyperandrogenic controls. After 3 days of transdermal E2 pretreatment (0.2 mg/day), subjects were admitted to the Clinical Research Unit (CRU) for a 24-hour frequent blood sampling protocol starting at 2000 h. (CRU admissions occurred no earlier than cycle day 7 in PCOS and between days 7 and 11 inclusive in controls.) At 0600 h, subjects received either 100 mg oral micronized P4 or placebo (PBO). In a subsequent menstrual cycle, subjects underwent an identical CRU protocol except that P4 was exchanged for PBO or vice versa. LH secretion was analyzed using Autodecon, a deconvolution program that provides estimates of LH pulse frequency, pulsatile LH secretion (amount of LH secreted as pulses), and basal (non-pulsatile) LH secretion. Results were analyzed using 2-period crossover design analysis of covariance. In both groups, neither LH pulse frequency nor basal LH secretion changed significantly with P4 (compared to changes with PBO). Mean LH increased with P4 in both groups—3.1-fold (95% CI, 2.4–4.0) in controls and 2.7-fold (95% CI, 2.1–3.5) in PCOS; in both groups, P4-related changes were significantly greater than PBO-related changes (Bonferroni-corrected p=0.012 and 0.010, respectively). In controls, pulsatile LH secretion increased 3.5-fold (95% CI, 2.3–5.2) with P4—significantly more than with PBO (p=0.029); while in PCOS, a 2.6-fold (95% CI, 1.8–3.9) increase with P4 was not significantly different from changes with PBO (p=0.911). In controls, mean FSH increased 2.0-fold (95% CI, 1.7–2.3) with P4—significantly more than with PBO (p=0.004); but in PCOS, a 1.5-fold (95% CI, 1.3–1.8) increase was not significantly different from changes with PBO (p=0.072). Despite the above, between-group (PCOS vs. controls) differences in P4-induced changes in pulsatile LH secretion and mean FSH were not formally (statistically) demonstrable. Between-group differences representing potential confounders included age (median 25.5 vs. 19.0 y; p=0.029), body mass index (29.9 vs. 21.8 kg/m2; p=0.006), and cycle day of CRU admissions (day 45.0 vs. 10.4 for P4 admissions; 30.0 vs. 10.0 for PBO admissions). In summary, these data suggest that P4-induced increases in pulsatile LH secretion and mean FSH may be blunted in PCOS compared to controls, which could contribute to ovulatory dysfunction in PCOS. However, our results do not confirm this possibility, and further study is needed.

Resolution of Abnormal Gonadotropin Secretion After Surgical Cure in Men With ACTH-Dependent Cushing Syndrome

Background: Hypothalamic-pituitary-gonadal (HPG) dysfunction is known to occur in male patients with Cushing Syndrome (CS) but the underlying mechanisms of HPG dysfunction remain unclear. We performed frequent blood sampling and GnRH stimulation tests to assess gonadotropin secretion in males before and after surgical cure of CS. Methods: We evaluated eight males age 32.5±12 years (mean ± SD) enrolled for surgical cure of ACTH-dependent CS at a tertiary care research center. Urinary free cortisol (UFC) was measured before and after surgery. Blood was sampled every 20 minutes for 24-hours with measurement of LH and FSH, before and 2 weeks to 160 weeks (median 80 weeks) after surgery. Patients received 100 μg of gonadotropin releasing hormone (GnRH) iv with measurement of LH and FSH at 0, 10 and 20 min before and 15, 30, 45, 60, 120 and 180 min after administration. Non-normative data was log-transformed before analysis using paired two-tail t-tests. A p-value of <0.05 was considered significant. Gonadotropin values are expressed in U/L as equivalents of the 2nd International Preparation of human menopausal gonadotropins with normal adult male ranges of 6-26 and 5-25 IU/L for LH and FSH, respectively. Results: Mean UFC was elevated ~ 6 times above the upper normal limit before surgery and returned to the reference range after surgery. 24-hour LH rose from 7.82 ± 1.48 preoperatively to 13.07 ± 2.96 IU/L (mean ± sem) after surgery (p=0.026) while mean FSH was unchanged (8.48 ± 1.51 vs 6.92 ± 1.29, p=0.37). LH pulse frequency, a marker of pulsatile GnRH secretion, recovered from a subnormal value of 6.88 ± 0.55 pulses/24 hr before to 12.13 ± 0.72 pulses/24 hr after surgery (p<0.0001) while LH pulse amplitude did not change (6.47 ± 1.21 vs 4.76 ± 0.88 IU/L, p=0.21). In response to GnRH there was a robust increase in LH that was not affected by curative surgery for CS (27.0 ± 7.2 vs 21.2 ± 7.6 IU/L; p=0.14) as was also the case for FSH (3.7 ± 1.2 vs 2.3 ± 0.5 IU/L; p=0.48), consistent with the absence of an effect of CS on gonadotroph responsiveness to LHRH. Conclusion: In men with Cushing Syndrome, hypogonadism is associated with inhibition of endogenous GnRH secretion but preservation of the pituitary response to GnRH. Hypothalamic suppression of the HPG axis is reversible after cure of CS.

Retrospective outcome in women aged 45 years and older undergoing natural cycle IVF treatment

Research questionIs natural cycle IVF treatment beneficial to middle-aged women with poor ovarian response? DesignRetrospective investigation of outcomes in women aged 45 years and older, who underwent natural cycle IVF treatment between 2009 and 2018 in a single assisted reproduction clinic with the aim of reporting several successful outcomes. ResultsIn total, 2408 IVF retrievals in women aged 45 years and older were included in this study. Mean serum FSH level on day 3 was 21.4 ± 12.5 (range: 0.3–93.7) IU/ml. One fresh cleavage-stage embryo was transferred in 37.4% (900/2408) of the initiated cycles. The overall clinical pregnancy rate and live birth rate per fresh embryo transfer were 2.8% (25/900) and 0.8% (7/900), respectively. Natural cycle IVF treatment led to seven successful deliveries during the period. All seven women who successfully delivered were poor ovarian responders who met the diagnostic Bologna criteria and, among them, three had elevated serum FSH levels on day 3 (range: 39.0–47.1 mIU/ml). All seven had full-term delivery, and no congenital abnormalities were observed in their infants. No significant difference was found in serum FSH level on day 3 between those with and without positive beta-HCG test results. ConclusionsThese findings suggest that natural cycle IVF treatment could be an option for older poor responders in countries that do not permit egg donation. Careful counselling is required, however, because of the low probability of live births after IVF in middle-aged women.