Number Of Antral Follicles Research Articles

Apigenin Improves Ovarian Dysfunction Induced by 4-Vinylcyclohexene Diepoxide via the AKT/FOXO3a Pathway.

Perimenopausal syndrome is a significant issue that disturbs women's metabolism, mood and quality of life. Apigenin (4',5,7-trihydroxyflavone) is a natural flavonoid that exhibits antioxidant, anti-inflammatory and anticancer effects. The present study aims to investigate the effect of apigenin on perimenopausal syndrome by combining bioinformatics analysis with in vivo experiments. The mouse model with perimenopausal syndrome was established using 4-vinylcyclohexene diepoxide (VCD) treatment. Apigenin alleviated VCD-induced disorder of estrous cycle and shrinkage of ovarian tissue. The reduction of anti-Muller hormone and the increase of follicle stimulation hormone and luteinizing hormone triggered by VCD were reversed by apigenin in a dose-dependent manner. Apigenin suppressed the VCD-induced decrease of primordial, primary, secondary and antral follicle number in ovarian tissue. Oxidative stress in ovarian tissue was activated by VCD treatment through increasing the reactive oxygen species production. High concentration of apigenin significantly reversed the alteration induced by VCD. Apigenin alleviated VCD-induced cell apoptosis through regulating Bax, Bcl-2, cleaved PARP1 and caspase-3. Furthermore, the phosphorylation of AKT and FOXO3a was inhibited by VCD and activated by apigenin in a dose-dependent manner. Collectively, apigenin effectively mitigates the ovarian dysfunction through suppressing oxidative stress and apoptosis via the AKT/FOXO3a signaling pathway.

High-altitude hypoxia exposure alters follicular metabolome and oocyte developmental potential in women.

To explore the effects of high-altitude hypoxia on the microenvironment of oocyte development and fertilization potential, we compared the metabolomic patterns of follicular fluid from women living in different altitude areas and traced their oocyte maturation and subsequent development. A total of 315 clinical cases were collected and divided into three groups according to their residence altitudes: 138 cases in low-altitude (< 2300m) group, 100 cases in middle-altitude (2300-2800m) group and 77 cases in high-altitude (> 2800m) group. The clinical outcomes were statistically estimated, including hormonal level, oocyte maturation, in vitro fertilization, and embryo development. Meanwhile, a metabolomic analysis was performed on the follicular fluid of women from different groups using ultra-high-performance liquid chromatography and high-resolution mass spectrometry and differential metabolites were analyzed through the KEGG pathway. The clinical data indicated that the physical condition and reproductive hormone secretion were similar among different groups. Although personalized gonadotropin-releasing hormone strategies were applied, the numbers of antral follicles and obtained oocytes were not impacted by the residence altitude change. In in vitro culture, the maturing rate, fertility rate and cleavage rate of high-altitude group were compared with the other groups. However, the rates of high-quality embryo, formative blastocyst, and available blastocyst were gradually decreased with the rise of residence altitude. Metabolome analysis identified 1193 metabolites in female follicular fluid. Differential analysis indicated that metabolic components in follicular fluid were remarkably changed with the elevation of residence altitude. These differential metabolites were closely related with amino acid metabolism, protein digestion and absorption, oocyte meiosis and steroid biosynthesis. The residence altitude alters the microenvironment of follicular fluid, which could damage the oocyte developmental potential. This study provides diagnostic basis and therapeutic targets for research on female oocyte and embryo development.

Ultrasound in in vitro fertilization programs

Ultrasound examination is an integral part of in vitro fertilization programs. This work shows that the antral follicle count compared to the level of antimüllerian hormone is a preferred marker for assessing the ovarian reserve, response to ovarian stimulation and the results of in vitro fertilization programs, while counting the antral follicle count can be carried out on any day of the menstrual cycle without reducing the informativeness of the study of the ovarian reserve, which is convenient for both patients and doctors. 3D transvaginal ultrasound allows automated assessment of the number of antral follicles with high accuracy and efficiency in less time than 2D transvaginal ultrasound. Using 3D transvaginal ultrasound and artificial intelligence, a threshold value of dominant follicle volume of 0.5 cm3 or greater was determined as a marker for predicting the number of mature oocytes in in vitro fertilization programs. The measurement of endometrial thickness and volume in combination with the determination of vascularization index, flow index, vascularization-flow index and systole-diastolic ratio in the uterine artery at transvaginal ultrasound allows predicting the results of in vitro fertilization programs. Modern possibilities of transabdominal oocyte aspiration using vaginal probe are presented.

Progesterone peak influences embryonic developmental morphokinetics on trigger day? A retrospective study

ObjectivePremature Progesterone Rise (PPR) is characterized by elevated serum progesterone concentrations either towards the end of the follicular phase or on the trigger day, surpassing a pre-defined threshold value. Aim of the study is to evaluate the impact of PPR exceeding 1.5 ng/ml at the time of hCG-trigger on embryo morphokinetic parameters and to identify predictive biomarkers of in IntraCytoplasmic Sperm Injection (ICSI) cycles outcomes.MethodsIt is a retrospective study including patients underwent ICSI cycles in the period 2020–2023. 58 patients were recruited in the study group showing P levels in the trigger day greater than or equal to 1.5 ng/ml. A matching control group of 58 patients with P levels below 1.5 ng/ml was after selected. The general characteristics of these patients, including age, Body Mass Index (BMI), antral follicle count (AFC), anti-Müllerian hormone (AMH) and follicle-stimulating hormone (FSH) levels, the type of infertility and smoking/non-smoking patients, were recorded on the day of their initial visit. Subsequently, data were collected regarding the number of eggs retrieved, mature eggs, successfully fertilized eggs, and embryos reaching the blastocyst stage. Additionally, the timing of embryonic development and the quality of obtained blastocysts, as assessed by the degree of expansion and the characteristics of the inner cell mass (ICM) and trophectoderm (TE), were evaluated using Time-Lapse technology.ResultsElevated P levels exceeding 1.5 ng/ml on the trigger day were directly associated with a significantly larger number of antral follicles, consequently leading to a higher count of retrieved eggs, mature eggs, successfully fertilized eggs and embryos reaching the blastocyst stage. Furthermore, the analysis of morphokinetic parameters indicated faster division times and a notably greater number of high-grade blastocysts in the study group compared to the control group.ConclusionsP levels ≥ 1.5 ng/ml on the trigger day did not negatively impact embryonic morphokinetic parameters, instead resulting in faster embryo development in the initial stages.

The impact of enhanced early postnatal nutrition on the histomorphological features of the ovaries in fat-tailed Sanjabi ewe lambs.

This study aimed to investigate whether providing additional ad libitum feeding could improve reproductive characteristics in prepubertal ewe lambs. Twelve Sanjabi ewe lambs, approximately one-month-old and with an average initial weight of 11.6 ± 1.3kg, were monitored over a 3-month period, which was divided into three distinct periods of 30 days each. The first period spanned from the lambs' age of one to two months, the second period covered their age range of two to three months, and the third period encompassed their age range of three to four months. The lambs were randomly assigned to either a control group (C) or a treatment group (T). The control group received a basal diet consisting of hay and milk, whereas the treatment group received an enhanced diet comprising hay, milk, and concentrate. Live weight (LW) and average daily gain (ADG) were measured weekly, and blood samples were collected from the jugular vein at days 30, 60, and 90 of the experiment to determine the concentrations of progesterone (P4) and estrogen (E2). Following the 90-day experiment, the lambs were humanely euthanized in accordance with established protocols. Subsequently, both ovaries from each animal were removed, weighed and subjected to a comprehensive histological analysis. The ovaries underwent fixation, embedding, and sectioning procedures, followed by staining with hematoxylin and eosin (H&E) to facilitate microscopic examination. Ovary microscopic sections from both C and T groups were examined to assess follicles at various developmental stages, including primordial follicle number (PreFN) and diameter (PriFD), primary follicle number (PriFN) and diameter (PriFD), secondary follicle number (SFN) and diameter (SFD), antral follicle number (AFN) and diameter (AFD), and granulosa layer thickness of secondary (GLTSF) and antral follicles (GLTAF). The impact of diet and three experimental periods on ADG and LW exhibited significant variations. There was a significant increase (P < 0.05) in LW in the T group compared to the T group, with noticeable differences in LW fluctuations observed during distinct periods. However, ADG was more prominent in the initial and second observation periods than in the third. The impact of diet and experimental periods on P4 and E2 concentrations was found to be negligible. The T group showed significant improvements (P < 0.05) in PreFD, PriFN, and GLTAF compared to the C group. This study suggests that ad libitum concentrate feeding may promote reproductive characteristics in prepubertal ewe lambs. Further research is needed to explore its potential impact on the age of puberty.

Yougui Pills Alleviates Diminished Ovarian Reserve Through Regulating Oxidative Stress and Apoptosis in Rats

Objective Yougui pills (YGP) have been used clinically to treat diminished ovarian reserve (DOR), but, whether YGP could have a palliative effect in the DOR rat model and the underlying mechanisms have not been clearly elucidated. This study aims to investigate the therapeutic effect and potential mechanisms of YGP in rats with cortisol-induced DOR. Methods The cortisol-induced DOR model was used to investigate the protective effects and mechanism of YGP on DOR. The DOR model was established by intramuscular injection of 25 mg/kg cortisol for 28 consecutive days. Meanwhile, the rats were weighed and administered intragastrically YGP or progynova. To evaluate the mitigating effect of YGP on DOR, pharmacodynamic indicators were assessed including body weight, estrous cycle, H&amp;E staining, and serum levels of AMH, FSH, LH, and cortisol. Oxidative stress (OS) and apoptosis pathways were further validated using Western blotting analysis. Result YGP significantly ameliorated the symptoms of DOR. Administration of YGP improved the ovarian index, estrous cycle, and body weight. After treatment with YGP, cortisol, FSH, and LH levels declined and AMH levels elevated. YGP increased the number of primary, secondary, and antral follicles, but decreased the number of atretic follicles. Mechanically, YGPtreated cortisol-induced DOR via the Keap1/Nrf2 signaling pathway which regulates OS. YGP also reversed the aberrant expression of apoptosis-related proteins Caspase-3, Bcl-2, and Bax. Conclusion YGP could reduce ovarian OS and apoptosis in cortisol-induced rats via regulating the Keap1/Nrf2 signaling pathway, thereby enhancing ovarian reserve function.

Assessment of ovarian cortex follicles in chemotherapy naïve and chemotherapy exposed patients.

Histological evaluation of ovarian tissue harvested as part of the attempt to preserve fertility might clarify the mechanism by which ovarian failure is caused. The purpose of this study was to compare the histologic appearance of ovarian tissue harvested for ovarian tissue cryopreservation (OTC) in chemotherapy naïve and chemotherapy exposed patients regarding the presence of follicles in different stages of development and to explore ovarian tissue histology in patients exposed to low- and high-cytotoxicity risk chemotherapy. A cohort of post-pubertal cancer survivors who underwent OTC between 1997 and 2018 was evaluated. An expert pathologist reviewed the pathology slides taken during OTC. The assessment included counting number of primordial, primary, secondary, and antral follicles. A comparison was made between chemotherapy naïve and chemotherapy exposed women and further between women who previously received low- versus high-risk gonadotoxic regimens. Seventy post-pubertal patients were included in the study. Seventeen (24.3%) received chemotherapy prior to OTC, and 53 (75.7%) were chemotherapy naïve at the time of OTC. A significant difference was found only in the number of secondary follicles, which was increased in chemotherapy naïve patients (2.81±4.26 vs. 0.88±1.18, respectively; P=0.005). Similar results were observed in a subgroup analysis of hematologic malignancies separately. Comparison of patients with previous exposure to chemotherapy revealed similar follicular appearance, except for the number of secondary follicles, which was higher in patients receiving low-risk compared to high-risk chemotherapy (1.40±1.28 vs. 0.14±0.35, respectively; P=0.006). The ovarian follicular pool at OTC appears comparable between chemotherapy naïve individuals and those post-exposure, as well as among patients receiving low versus high-risk gonadotoxic regimens, with the exception of secondary follicles, which are presented in increased numbers in chemotherapy naïve and those exposed to low-risk gonadotoxic chemotherapy.

The effect of norepinephrine on ovarian dysfunction by mediating ferroptosis in mice model.

Studies have shown that stress is associated with ovarian dysfunction. Norepinephrine (NE), a classic stress hormone involved in the stress response, is less recognized for its role in ovarian function. In this study, an NE-treated mouse model is induced by intraperitoneal injection of NE for 4 weeks. Compared with normal control mice, NE-treated mice show disturbances in the estrous cycle, decreased levels of anti-Mullerian hormone (AMH) and estradiol (E2), and increased level of follicle-stimulating hormone (FSH). Additionally, the numbers of primordial follicles, primary follicles, secondary follicles, and antral follicles are decreased, whereas the number of atretic follicles is increased in NE-treated mice, indicating NE-induced ovarian dysfunction. RNA sequencing further reveals that genes associated with ferroptosis are significantly enriched in NE-treated ovarian tissues. Concurrently, the levels of reactive oxygen species (ROS), ferrous ions, and malondialdehyde (MDA) are increased, whereas the expression level of glutathione peroxidase 4 (GPX4) is decreased. To elucidate the mechanism of NE-induced ferroptosis in ovaries and the potential reversal by Coenzyme Q10 (CoQ10), an antioxidant, we conduct both in vitro and in vivo experiments. In vitro, the granulosa cell line KGN, when treated with NE, shows decreased cell viability, reduced expression of GPX4, elevated levels of ferrous ion and ROS, and increased MDA level. However, these NE-induced changes are reversed by the addition of CoQ10. Compared with the NE group, the NE-treated mice supplemented with CoQ10 present increased GPX4 level and decreased iron, ROS, and MDA levels. Moreover, the differential expression of genes associated with ferroptosis induced by NE is ameliorated by CoQ10 in NE-treated mice. Additionally, CoQ10 improves ovarian function, as evidenced by increased ovarian weight, more regular estrous cycles, and an increase in follicles at various stages of growth in NE-treated mice. In conclusion, NE induces ovarian dysfunction by triggering ferroptosis in ovarian tissues, and CoQ10 represents a promising approach for protecting reproductive function by inhibiting ferroptosis.

The Effects of L-Tartaric Acid on Ovarian Histostereological and Serum Hormonal Analysis in an Animal Model of Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is the most common endocrine-related reproductive disorder in women of reproductive age, accompanied by both the impairment of female fecundity and a risk of metabolic disorders. PCOS is emphasized as a worldwide concern due to its unknown etiology and lack of specific medications. The current study aimed to evaluate the effects of L-tartaric acid, an abundantly occurring compound in fruits, on the histostereological and hormonal changes caused by PCOS. Forty adult Sprague Dawley rats were randomly divided into four groups including controls (no intervention), Tartaric acid (40mg/Kg/day from day 21 onwards for 39 days), PCOS (21 days letrozole and then normal saline orally for 39 days), and PCOS + Tartaric acid. After treatments, the ovarian histostereological analysis as well as the level of reproductive hormones including luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol, progesterone, and testosterone was measured. PCOS caused a significant decrease in the number of unilaminar, multilaminar, antral, and graafian follicles and increased follicular atresia (p-value < 0.001). Moreover, the weight and volume of ovarian tissue and related structures including cortex, medulla, and cysts increased significantly (p-value < 0.0001). However, corpus luteum volume was significantly decreased (p-value < 0.001). Although significant differences were found in some parameters with the control group (p-value < 0.05), the administration of tartaric acid restored the pathological effects of PCOS on the ovarian histostructure. Furthermore, tartaric acid improved the serum levels of LH, estradiol, progesterone, and testosterone (p-value < 0.05). The obtained findings may suggest tartaric acid as a novel strategy for PCOS management, although further studies are necessary.

Ovarian and Uterine Morphology in Minipuberty: Associations with Reproductive Hormones. A COPANA Study of 302 girls.

Female minipuberty is characterized by complex dynamics of circulating reproductive hormones, while the relationship between ovarian and uterine morphology and reproductive hormones remains to be elucidated. To investigate the association between reproductive hormones and ovarian as well as uterine morphology by transabdominal ultrasound (TAUS) at minipuberty. Secondary analysis from The Copenhagen Analgesic Study (COPANA) (ClinicalTrials.gov NCT04369222). Healthy infant girls (n=302, age 3.4 months (0.4) mean (±SD): mamma tissue diameter (mm), n=300. TAUS: uterine volume (n=230), endometrial thickness (n=255), ovarian volume, antral follicle count (total/2-4mm/≥5mm) (n=203).Blood samples (n=269/302=89%): AMH, FSH, LH, inhibin B (immunoassays), progesterone (PROG), androstenedione (Adione), testosterone (T), estrone (E1), estradiol (E2) (LC/MS-MS).Statistics: Pearson/Spearman´s correlation coefficient (parametric/non-parametric data). Total follicle count correlated positively with ovarian volume (r= 0.606, p<0.001), AMH (r=0.378, p<0.001), inhibin B (r=0.251, p<0.001), and negatively with FSH concentrations (r=-0.327, p<0.001). Larger follicles (≥5mm) correlated positively with AMH (r=0.264, p<0.001), inhibin B (0.230, p=0.002), E1 (r=0.209, p=0.004), E2 (r=0.269, p<0.001), PROG (r=0.160, p=0.031) and T (r=0.210, p=0.004) and negatively with FSH (r=-0.183, p=0.015). Circulating E1 and E2 levels correlated with the size of estrogen-responsive tissue sizes: E2 vs. uterine volume (r=0.134, p=0.054), E2 vs. endometrial thickness (r=0.155, p=0.020), E1 and E2 vs. mammary tissue diameter (r=0.213 and r=0.198, respectively, both p < 0.001). Correlations between reproductive hormones and the number of antral follicles suggest that negative feedback in the female HPG axis is established during minipuberty, with ovarian activity promoting uterine and glandular breast tissue growth We provide normative data of infant ovarian- and uterine morphology directly implementable to a clinical setting.

Stereological and biochemical effects of thymoquinone on ovarian tissue toxicity induced by silver nanoparticles in NMRI mice: An experimental study.

The toxicity of silver nanoparticles (AgNPs) has been proven in the female reproductive system. Thymoquinone (TQ) is a natural antioxidant and bioactive component of Nigella sativa. We evaluated the efficacy of TQ on ovarian tissue following toxicity induced by AgNPs in female mice. 24 female NMRI mice (5-6 wk, an average weight of 33 gr) were randomly divided into 4 groups (n = 6/each): control, AgNPs (500 mg/kg, gavage), TQ (2.5 mg/kg, intraperitoneal injection), and TQ+AgNPs. Mice were treated every day for 35 days. Serum levels of malondialdehyde (MDA), total antioxidant capacity (TAC), luteinizing hormone, and follicle-stimulating hormone were measured. The optical disector and stereological techniques were utilized to estimate the follicular count, their volume at different developmental stages, and the structure of ovarian tissue. In the AgNPs group, the serum concentrations of TAC (p = 0.01), luteinizing hormone (p 0.001), follicle-stimulating hormone, the volume of corpus luteum (p 0.001), and the number of follicles decreased significantly compared to the control group. Nevertheless, AgNPs significantly increased the MDA level. In the TQ+AgNPs group compared to the AgNPs group, a significant decrease in MDA level (p 0.001) and a significant improvement in TAC (p = 0.03), and hormonal levels, the number of primary, preantral, and antral follicles (p = 0.04), and the volume of corpus luteum (p = 0.01) were observed. TQ improved the number of follicles by reducing oxidative stress and lipid peroxidation in AgNPs-damaged ovarian tissue.

Paeoniflorin Promotes Ovarian Development in Mice by Activating Mitophagy and Preventing Oxidative Stress.

During the development of animal organs, various adverse stimuli or toxic environments can induce oxidative stress and delay ovarian development. Paeoniflorin (PF), the main active ingredient of the traditional Chinese herb Paeonia lactiflora Pall., has protective effects on various diseases by preventing oxidative stress. However, the mechanism by which PF attenuates oxidative damage in mouse ovaries remains unclear. We evaluated the protective effects of PF on ovaries in an H2O2-induced mouse oxidative stress model. The H2O2-induced mouse ovarian oxidative stress model was used to explore the protective effect of PF on ovarian development. Histology and follicular development were observed. We then detected related indicators of cell apoptosis, oxidative stress, and autophagy in mouse ovaries. We found that PF inhibited H2O2-induced ovarian cell apoptosis and ferroptosis and promoted granulosa cell proliferation. PF prevented oxidative stress by increasing nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression levels. In addition, the autophagic flux of ovarian cells was activated and was accompanied by increased lysosomal biogenesis. Moreover, PF-mediated autophagy was involved in clearing mitochondria damaged by H2O2. Importantly, PF administration significantly increased the number of primordial follicles, primary follicles, secondary follicles, and antral follicles. PF administration improved ovarian sizes compared with the H2O2 group. The present study suggested that PF administration reversed H2O2-induced ovarian developmental delay and promoted follicle development. PF-activated mitophagy is crucial for preventing oxidative stress and improving mitochondrial quality.

Risk factors for diminished ovarian reserve in women: Current state of the problem

Ovarian reserve is the basis of female fertility. The main markers of ovarian reserve are the level of anti-Mullerian hormone and the number of antral follicles. In addition to the natural age-related loss of follicles, many women experience a premature diminished ovarian reserve associated with a number of factors. This can be caused by both various diseases and environmental factors, lifestyle, and social aspects.The aim of this review was to examine the influence of external factors on the ovarian reserve and women fertility. A systematic analysis of data from modern scientific literature, domestic and foreign sources was carried out. The search involved such resources as PubMed, MEDLINE, Science Direct, eLibrary, Scopus, Cyberleninka. A detailed analysis of the influence of environmental pollution, lifestyle (sleep, nutrition, physical activity), previous surgeries, bad habits, obesity, psychological and social factors on the ovarian reserve and reproductive function of women was carried out. Significantly diminished ovarian reserve was noted with low sleep quality, excessive physical activity, and an unbalanced diet poor with animal proteins. Regular consumption of alcohol, smoking and exposure to certain chemical environmental pollutants cause premature follicle apoptosis and the onset of menopause. Circadian dysrhythmia, chronic stress and obesity can lead to the ovarian menstrual cycle disorders and the development of infertility in women. Previous parovarium surgeries are a significant risk factor for diminished ovarian reserve. Further population-based studies are needed to determine the precise mechanisms of influence of various factors on female fertility.

Study of Sonographic Indicators of Ovarian Reserves in Women With WHO-Defined Anovulatory Disorders.

Depending on the age of the wife or female partner, infertility is defined as the inability of a couple to achieve conception after 12 months if the age is ≤35 years and six months if the age is ≥35 years. About half of female infertility is attributed to ovulatory dysfunction, and the rest is either due to tubal and pelvic pathology or problems such as thyroid disease or anatomic abnormalities. Ovulatory disorders are one of the major factors for infertility problems in couples. Ultrasound monitoring of ovarian follicle growth plays a pivotal role in infertility treatment. The total number of small antral follicles in both ovaries is an important predictive parameter of the ovarian induction cycle, whereas ovarian volume is the parameter most easily assessed by ultrasound. The purpose of this study is to analyze the sonographic indicators of infertility (antral follicle count and ovarian volume) in four World Health Organization (WHO)-defined anovulatory groups and compare the same with that of healthy fertile women.

In vitro fertilization results of GNRH antagonists and medroxyprogesterone acetate used to prevent premature LH surge during ovarian hyperstimulation

The aim of this study was to evaluate the effects of medroxyprogesterone acetate (MPA) treatment in comparison to those of gonadotropin releasing hormone (GnRH) antagonists for the prevention of premature luteinizing hormone surges during controlled ovarian hyperstimulation (OS) and the impact of these effects on developing embryos and pregnancy outcomes. Data from 757 cycles of GnRH antagonist treatment and 756 cycles of MPA treatment were evaluated at the Akdeniz University Faculty of Medicine Assisted Reproductive Treatment Center between October 2018 and April 2022. Patient records were obtained from the electronic database of the centre and analysed. In our centre, GnRH antagonist protocols were used between 2018 and 2020, and MPA protocols were used between 2020 and 2022. We chose our study population by year. Our study is a comparative retrospective study. All methods in this study were performed in accordance with the relevant guidelines and regulations. Patients using MPA were significantly older (33.9 ± 5.6 vs. 32.6 ± 5.6, p < 0.001) and had a lower number of antral follicles (AFC) (10.7 ± 8.6 vs. 11.9 ± 10.8, p = 0.007) than those using GnRH antagonists. Both MPA (2.9%) and GnRH antagonists (2.2%) had similar effectiveness in preventing premature ovulation (p = 0.415). There was no significant difference between the two groups in terms of the number of total developed embryos (1.3 ± 1.3 vs. 1.2 ± 1.2, p = 0.765). There was no significant difference in the clinical pregnancy rates with the first ET (%35.4 vs. %30.1, p = 0.074), per total number of transfers (35.3% vs. 30.1%, p = 0.077). MPA was found to be effective at preventing premature ovulation during OS treatment, and the incidence of developing embryo and pregnancy outcomes in patients using MPA were similar to those in patients using GnRH antagonists. Therefore, the use of MPA instead of GnRH antagonists during OS may be a viable alternative for patients not scheduled for fresh ET.

P-705 Telomere length of white blood cells is a non-invasive marker of reproductive aging in women

Abstract Study question Can leukocyte telomere length be a marker of ovarian decline? Summary answer The significant correlation found between leukocyte and cumulus cell telomere lengths supports the use of leukocyte telomere length (TL) as a marker of ovarian aging. What is known already The telomere-based theory of reproductive aging proposes that age-related ovarian dysfunction is a consequence of progressive telomere shortening from fetal oogenesis to the adult ovary, compromising genome stability and proper chromosome segregation. Telomere length in cumulus cells (CCTL) is known to be a biomarker of oocyte and embryo quality, but its assessment is not practical. Hence, we previously proposed leukocyte telomere length (LTL) as an alternative to directly measuring telomere length in follicular cells, but so far it has only been assessed in samples of moderate size and homogeneous populations without consistent results. Study design, size, duration This prospective, non-interventional cohort study involved 300 women aged 18 to 46 years undergoing controlled ovarian stimulation over a 24-month period. The cohort included 169 egg donors and 131 IVF patients. Oocyte retrieval was performed 36 hours after maturation induction, with cumulus cells collected during oocyte stripping and blood samples taken via peripheral venous access. Participants/materials, setting, methods Genomic DNA was isolated from leukocytes and cumulus cells of all subjects for TL measurements. Relative TL was determined using a SYBR-Green quantitative real-time PCR protocol. A Taqman assay for the multicopy gene Alu was used to normalize the measurements. A Pearson’s correlation test was used to analyze the correlation between CCTL and LTL, as well as between TL and age, antral follicle count, Anti-Müllerian hormone, and number of mature oocytes retrieved. Main results and the role of chance Mean age of the subjects was 30.70±7.61 years. Anti-Müllerian hormone (AMH) mean level was 2.60±1.87 ng/ml, antral follicle count (AFC) was 17.8±7.4 and the number of mature oocytes was 14.3±9.3. A significant correlation was observed between CCTL and LTL (R = 0.6, p-value=2.2E-16). Notably, the relationship between leukocyte and cumulus cell telomere lengths was stronger in the 97 participants over 35 years (R = 0.67, p-value=1.55E-13), especially those between 39 to 46 years (R = 0.70, p-value=1.19E-12). Telomere length was inversely correlated with age in both leukocytes and cumulus cells (R=-0.23, p-value=6.98E-05; R=-0.4, p-value=2.38E-05), meaning the older the subject, the shorter the telomeres in both cells. Furthermore, longer telomeres in leukocytes and cumulus cells were significantly correlated with higher AFC (R = 0.20, p-value=0.0006; R = 0.25, p-value=1.83E-05), and more mature oocytes retrieved (R = 0.16, p-value=0.0043; R = 0.27, p-value=2.83E-06). However, we did not find any association between AMH levels and LTL or CCTL (R = 0.07, p-value=0.2666; R = 0.07, p-value=0.2747). Limitations, reasons for caution The method used to measure the average telomere length from pooled leukocyte samples may not reflect individual cell variability. Additionally, it allows for relative but not absolute telomere length comparisons. Wider implications of the findings This study is the first to demonstrate in a large and heterogeneous population that LTL could be a viable marker for ovarian aging by reflecting follicular cell telomere length. This finding may provide a valuable tool for assessing and monitoring the reproductive health of IVF patients. Trial registration number Not applicable

P-646 Prognostic statistical model of age-specific anti-Müllerian hormone (AMH) decrease based on hormone values of 23,528 women

Abstract Study question The analysis of anti-Müllerian (AMH) levels, can a useful statistical model be constructed to predict the rate of decrease of AMH levels at different ages? Summary answer A determination of AMH levels can be an objective marker of a ovarian age and a prognostic marker of the rate of ovarian function decline. What is known already The anti-Müllerian hormone is a dimeric glycoprotein produced by granulosa cells of the antral follicles in the ovaries, and AMH participates in the formation of the dominant follicle during folliculogenesis. Serum AMH levels are relatively constant and show little variability between and within the menstrual cycle. The level of serum AMH is proportional to the number of antral follicles and therefore most suitable for determining ovarian reserve. For this reason, AMH is a convenient marker for the biological age of the ovary and for the response to controlled ovarian stimulation (COS). Study design, size, duration A nationwide cross-sectional retrospective study was conducted between 2017 and 2023 and included 23,528 women the ages of 20 - 50, in which AMH levels were measured once. We performed an analysis of the different AMH levels at different ages and constructed a predictive statistical model for the rate of AMH decline. Results were analyzed in subgroups with AMH values &amp;lt; 1 ng/mL, between 1 and 3,5 ng/mL, and &amp;gt; 3,5 ng/mL, at one-year intervals. Participants/materials, setting, methods All women who visited the laboratory for AMH testing were included in the analysis. The samples were collected in 38 points throughout the country and the analysis was carried out in one centralized laboratory using the Ansh Labs (Webster, TX, USA) manual assays, including the ultra-sensitive AMH enzyme-linked immunosorbent assay (ELISA), with a management range of 0,08 - 14,2 ng/ml. The intra-assay variations were 1,97% and 4,00%. The inter-assay variations were 4,63% and 1,98%, respectively. Main results and the role of chance When analyzing all samples in each consecutive year, we found that the rate of AMH decline was approximately linear and inversely proportional to the woman’s age. However, when analyzing the results for the rate of AMH reduction in the different subgroups, we found out that: In women with AMH &amp;lt; 1 ng/ml, the rate of decline with advancing age changes as follows: until age 34, the decline in AMH is very slow, between ages 35 and 45 there is a rapid acceleration of the process.. In women with AMH between 1 and 3,5 ng/ml, the rate of decline with age was approximately constant. In women with AMH &amp;gt; 3,5 ng/ml, the rate of hormone decline is approximately constant but the year-on-year decline rate is decreasing. Based on our analysis and clinical experience, it is our opinion that the rate of AMH decline for the low AMH group has a non-linear characteristic, while ingeneral different groups exhibit different dynamics for the rate of decline. On this basis, we created a statistical model to predict the decrease in AMH levels at different ages. Limitations, reasons for caution Women of only one ethnicity were included in the study. The studied group is heterogeneous, which does not exclude the inclusion of women with a certain pathology and infertility. In some cases, a single determination of AMH could not predict individual differences in rates of hormone reduction. Wider implications of the findings Determination of reference values for AMH at different ages, based on the large volume of examined samples may facilitate the application of AMH in clinical practice. Based on our statistical analysis, creating a prognostic model for decreasing ovarian function and time to menopause appears feasible. Trial registration number NA

Slit1 inhibits ovarian follicle development and female fertility in mice†.

Previous in vitro studies have suggested that SLIT ligands could play roles in regulating ovarian granulosa cell proliferation and gene expression, as well as luteolysis. However, no in vivo study of Slit gene function has been conducted to date. Here, we investigated the potential role of Slit1 in ovarian biology using a Slit1-null mouse model. Female Slit1-null mice were found to produce larger litters than their wild-type counterparts due to increased ovulation rates. Increased ovarian weights in Slit1-null animals were found to be due to the presence of greater numbers of healthy antral follicles with similar numbers of atretic ones, suggesting both an increased rate of follicle recruitment and a decreased rate of atresia. Consistent with this, treatment of cultured granulosa cells with exogenous SLIT1 induced apoptosis in presence or absence of follicle-stimulating hormone, but had no effect on cell proliferation. Although few alterations in the messenger RNA levels of follicle-stimulating hormone-responsive genes were noted in granulosa cells of Slit1-null mice, luteinizing hormone target gene mRNA levels were greatly increased. Finally, increased phospho-AKT levels were found in granulosa cells isolated from Slit1-null mice, and SLIT1 pretreatment of cultured granulosa cells inhibited the ability of both follicle-stimulating hormone and luteinizing hormone to increase AKT phosphorylation, suggesting a mechanism whereby SLIT1 could antagonize gonadotropin signaling. These findings therefore represent the first evidence for a physiological role of a SLIT ligand in the ovary, and define Slit1 as a novel autocrine/paracrine regulator of follicle development.

Weightlessness leads to an increase granulosa cells in the growing follicle

The participation of women in space programs of increasing flight duration requires research of their reproductive system from the perspective of subsequent childbearing and healthy aging. For the first time, we present hormonal and structural data on the dynamics of recovery after a 157-day space flight in a woman of reproductive age. There were no clinically significant changes in the reproductive system, but detailed analysis shows that weightlessness leads to an increase in the proportion of early antral follicles and granulosa cells in large antral follicles. Returning to Earth’s gravity reduces the number and diameter of early antral follicles.

Sucralose triggers insulin resistance leading to follicular dysplasia in mice

Sucralose, the extensively utilized sweetener, might lead to metabolic disorders with prolonged consumption, but it remains uncertain if sucralose has any impact on female reproductive health. We incorporated sucralose into drinking water and observed food intake, body weight, estrous cycle, follicular development, serum hormones, and insulin sensitivity of mice. The mice did not experience any changes in their food intake or body weight after consuming sucralose. However, they displayed irregularities in the estrous cycle, marked by a reduced count of primordial, primary, and secondary follicles, coupled with a significant increase in the number of antral follicles. There was a decline in follicle-stimulating hormone (FSH), estradiol (E2), and progesterone (P4) levels, while testosterone (T) and luteinizing hormone (LH) levels surged, leading to a notable elevation in the LH / FSH ratio. Sucralose also induced insulin resistance, as evidenced by elevated insulin levels and impaired insulin tolerance, which responded to an increase in bacterial-derived serum endotoxin. By eliminating insulin resistance with rosiglitazone (RSG), eradicating intestinal flora-derived endotoxins with neomycin (NEO), or enhancing intestinal barrier function with indole-3-carbinol (I3C), the abnormalities in estrous cycle, disruptions in follicular development, hormonal imbalances and elevation in serum endotoxins induced by sucralose were successfully reversed. The present study indicates that sucralose-induced follicular dysplasia in mice is probably related to impaired intestinal permeability, infiltration of endotoxins, initiation of systemic inflammation, and insulin resistance.