Ovarian Status Research Articles

Phenology and social status of the endemic Sable Island Sweat Bee, Lasioglossum sablense

Lasioglossum (Dialictus) sablense Gibbs, 2010 (Hymenoptera: Halictidae) is endemic to Sable Island, an isolated sandbar located about 160 km east of Nova Scotia, Canada. Lasioglossum sablense is currently listed as Threatened due to its restricted geographic distribution, and promoting its conservation requires detailed information about nesting phenology, behaviour, and other life history details. We combined measurements of body size, wear, and ovarian status from adult females collected from flowers in 2016 and 2017 with measurements and behavioural observations of foragers collected at the Sable Island Station in 2019 and 2022, to compile a preliminary description of flight phenology and social behaviour. Like many members of its genus, L. sablense exhibited a diphasic life history. Phase 1 began when large, fertilized adult females emerged from hibernation, began nest and brood cell construction, and foraged to provision Brood 1, which comprised both daughters and sons. Most nests initiated during Phase 1 reactivated during Phase 2, with adult Brood 1 daughters foraging to provision Brood 2. Observations of bees at nest entrances demonstrated that some reactivated nests contained multiple adult females. Phase 1 foragers were larger, less worn, and had marginally higher levels of ovarian development than Phase 2 foragers. Five large females collected during Phase 2 had high levels of wear, suggesting they were long-lived Phase 1 foundresses that resumed foraging during Phase 2. Taken together, these observations suggest a mix of univoltine (single generation) and bivoltine (two generation) life histories, as well as a mix of solitary and social behaviour in L. sablense.

6962 Natural History of Menstrual Pain and Associated Risk Factors in Early Adolescence

Abstract Disclosure: M. Sofia: None. R. Modarelli: None. S. Molsberry: None. S. Denslow: None. D. Zaccaro: None. C. Kamoun: None. N.D. Shaw: None. Background: Primary dysmenorrhea (PD), or menstrual pain of uterine origin without pelvic pathology, affects up to 93% of adolescents and is a leading cause of school and work absenteeism. The theory that PD only occurs when progesterone levels fall in the late-luteal phase, triggering production of uterine prostaglandins, has dominated the literature. However, recent studies identifying PD among girls with anovulatory cycles cast doubt on this theory. We took advantage of an ongoing longitudinal study of healthy early post-menarchal girls to test the hypothesis that ovulation is associated with increased PD and to explore other participant factors associated with PD. Methods: Participants contributed daily urine samples to measure Cr-corrected LH, estrone conjugates, and pregnanediol-3-glucuronide [PdG], completed menstrual diaries, and reported dysmenorrhea symptoms using the 24-item Menstrual Symptom Questionnaire (MSQ) approximately biannually. A cycle was defined as ovulatory if peak PdG >2500 ng/mg Cr. Scores were natural log-transformed and modeled to assess associations with gynecologic age, ovulation, anxiety, depression, and physical activity exposures using generalized estimating equations with a normal distribution and autoregressive covariance structure. Models were adjusted for age at menarche, baseline BMI Z-score, race, ethnicity, maternal age at menarche, parental education and employment, and gynecologic age (ovulation models only). Results: The 43 participants were 12.38 ± 1.06 years old (mean ± SD) with a gynecologic age of 2.99 ± 2.22 months. The majority were non-Hispanic White and of normal weight. Average maternal age at menarche was 12.64 ± 1.09 years. Among parents of participants, most were both at least college-educated and nearly 50% were both employed. Participants completed 155 MSQs (1-9/participant). Dysmenorrhea scores increased over time (scores 1.14x higher for every 1-year increase in gynecologic age, p<0.001). Dysmenorrhea scores were 1.12x higher (p=0.03) if the preceding cycle was ovulatory vs. anovulatory and 1.11x higher if all cycles in the preceding 6 months were ovulatory vs all anovulatory (p=0.11). Gynecologic age remained significant in models with ovulation-based exposures, and maternal age at menarche was inversely associated with dysmenorrhea score (beta -0.08, p=0.04) in the model with gynecologic age as the exposure. Conclusion: In early-postmenarchal girls, dysmenorrhea scores were higher when associated with ovulatory cycles but also increased over time independent of ovulatory status, suggesting a pathophysiologic role for progesterone and other unknown factors. There was no association between age at menarche, BMI, race/ethnicity, and dysmenorrhea scores whereas maternal age at menarche was inversely associated with dysmenorrhea scores. Presentation: 6/1/2024

Ovum Pick-Up and Ovaries Characterization of Black Bengal Goat from Slaughterhouse

Black Bengal goat is the only indigenous breed in Bangladesh. These are small-sized goats renowned for their high-quality meat, skin, and adaptability to adverse environmental conditions. To conserve the originality of this species, it is mandatory to develop an effective methodology for their germ cells to be harvested after death. Therefore, the study was conducted to assess the ovarian status and evaluate the most suitable method of ovum pick-up as well as the characterization of ovum from slaughterhouse specimens. A total of 182 ovaries were collected from local slaughterhouses from June 2022 to July 2023 and grossly examined to assess their status. Two methods were used to retrieve oocytes from the ovaries: aspiration and slicing. The grading of the ovum was done based on the germ layer present surrounding the oocytes. From 182 ovaries, the rates of corpus luteum (CL), tumor, and cyst were found to be 14.29%, 6.59%, and 27.47%, respectively. In the aspiration method, the significantly highest percentage of recovery rate (38.89%) and grade A oocytes (51.06%) were found within 1–3 hours of slaughtering. In the slicing method, the significantly highest percentage of recovery rate (19.57%) and grade A (60.61%) oocytes were found within 1–3 hours of slaughtering. The recovery rate was significantly higher (34.1%) in the aspiration method than in the slicing method (17.1%). There was no significant relationship in oocyte quality between the two methods. The recovery rate was not significantly dependent on the presence of CL. In conclusion, the findings from this study may be useful for assessing the quality of oocytes to preserve the germ cells and utilize them for in vitro fertilization for conserving Black Bengal goat.

183 Comparison of presynchronization and OvSynch protocols following fixed-time breeding in lactating dairy cattle

Abstract Current study was carried out to compare two presynch-OvSynch protocols with standard OvSynch protocol in postpartum Holstein dairy cattle. Postpartum cows (n = 473) were randomly divided into one of the three protocols: I) G7G-OvSynch (n = 159), cows were subjected to PGF2α followed by GnRH 48 h later. After 7 d post-GnRH injection, standard OvSynch protocol was introduced, II) G7GM-OvSynch (n = 162), cows were subjected to the same protocol as in group I) except one extra PGF2α at 12 h interval in Ovsynch part, and III) standard OvSynch protocol (n = 152). The progesterone (P4) profile was subjected to d 30 and d 60 post FTAI. Ovarian status was monitored at the 1st GnRH, PG, and 2nd GnRH injection of the breeding part. Pregnancy/ AI was diagnosed on d 30,60, and 90 post-FTAI, and pregnancy loss was also on d 60 and 90 post-FTAI. Pregnancy data were analyzed using the Chi-square test and plasma P4 profile by ANOVA using SAS. Ovulatory follicle diameter (mean ± SD) was 15.44 ± 1.24 in G7G-Ovsynch, 15.21 ± 1.36 in G7GM-OvSynch, and 14.90 ± 1.02 in OvSynch, respectively (P > 0.05). Plasma P4 profile (ng/mL; Mean ± SD) on d 30 post TAI was 6.50 ± 1.39,7.01 ± 1.13 and in 6.49 ± 1.32 G7G-OvSynch, G7GM-Ovsynch and OvSynch groups, respectively (P = 0.09). On d 60 post TAI, P4 profile was recorded to be 7.75 ± 1.20, 7.58 ± 1.36 and 6.80 ± 0.96 in G7G-OvSynch, G7GM-OvSynch and OvSynch, respectively (P < 0.05). P/AI on d 30 was 51.57% (83/159), 57.41% (93/162) and 42.76% (65/152) G7G-Ovsynch, G7GM-OvSynch, and OvSynch, respectively (P = 0.033). Similarly, AI on d 60 and d 90 post FTAI was 44.65% (1/159):51.85% (84/162), 38.16% (58/152): 43.40% (69/159) and 51.23% (83/162):36.18% (55/152) in G7G-Ovsynch, G7GM-OvSynch, and OvSynch protocols (P = 0.051: 0.027), respectively. Pregnancy loss on d 60 and d 90 post FTAI was 13.41%: 2.90%, 9.68%: 1.20% and 10.77%: 5.20% in G7G-Ovsynch, G7GM-OvSynch, and OvSynch, respectively (P = 0.73: 0.35). Although statistically non-significant (P = 0.56), overall pregnancy loss on d 90 post-FTAI was also numerically greater in the G7G-OvSynch (15.85%) protocol as compared with G7GM-OvSynch (10.75%) and OvSynch (15.38%). We concluded that the G7GM-OvSynch protocol has resulted in an increased P/AI, P4 profile in comparison with G7G-OvSynch and OvSynch. Similarly, pregnancy loss was also less in the G7GM-Ovsynch protocol, which makes it a protocol of choice for postpartum dairy cows.

A retrospective analysis of sheep generated by somatic cell nuclear transfer

Somatic cell nuclear transfer (SCNT) is one of the primary methods for production of genetically engineered sheep, which allows for gene editing or transgene introduction in somatic cells. The use of SCNT eliminates the risk of genetic mosaicism in embryos and animals that is commonly observed after zygote micromanipulations. This retrospective analysis of SCNT in sheep performed at Utah State University, spanning from 2016 to 2021, examined parameters that may impact pregnancy and full-term development, including donor oocytes (donor age), donor cell lines, SCNT parameters (time of oocyte activation following SCNT, number of transferred embryos, in vitro maturation and culture conditions), and recipients (surgical number and ovulatory status), as well as factors that may correlate with large offspring syndrome or abnormal offspring syndrome (LOS/AOS) in the fetuses and lambs. Our findings indicated that compared to prepubertal oocytes, the SCNT embryos produced from adult sheep oocytes had comparable in vitro maturation rates, pregnancy and full-term development rates, as well as SCNT efficiency. In addition, earlier activation time of SCNT embryos (e.g. 24–26 h post maturation) was correlated to the early pregnancy loss rate, full-term rate, and SCNT efficiency. Compared to our standard serum-containing medium, commercial serum-free culture medium showed a positive correlation with the full-term development of sheep SCNT embryos. Transferring 15–30 embryos per recipient resulted in consistently good pregnancy rates. Surgical numbers and ovulatory status (having at least one follicle between 6 and 12 mm in size or a corpus hemorrhagicum (CH)) of recipients did not affect pregnancy and full-term development rates. In summary, this retrospective analysis identified parameters for improving pregnancy and full-term development of SCNT embryos in sheep.

Investigating the Impact of Intracerebroventricular Streptozotocin on Female Rats with and without Ovaries: Implications for Alzheimer's Disease.

To contribute to research on female models of Alzheimer's disease (AD), our aim was to study the effect of intracerebroventricular (ICV) injection of streptozotocin (STZ) in female rats, and to evaluate a potential neuroprotective action of ovarian steroids against STZ. Female rats were either ovariectomized (OVX) or kept with ovaries (Sham) two weeks before ICV injections. Animals were injected with either vehicle (artificial cerebrospinal fluid, aCSF) or STZ (3mg/kg) and separated into four experimental groups: Sham + aCSF, Sham + STZ, OVX + aCSF and OVX + STZ. Nineteen days post-injection, we assessed different behavioral aspects: burying, anxiety and exploration, object recognition memory, spatial memory, and depressive-like behavior. Immunohistochemistry and Immunoblot analyses were performed in the hippocampus to examine changes in AD-related proteins and neuronal and microglial populations. STZ affected burying and exploratory behavior depending on ovarian status, and impaired recognition but not spatial memory. STZ and ovariectomy increased depressive-like behavior. Interestingly, STZ did not alter the expression of β-amyloid peptide or Tau phosphorylated forms. STZ affected the neuronal population from the Dentate Gyrus, where immature neurons were more vulnerable to STZ in OVX rats. Regarding microglia, STZ increased reactive cells, and the OVX + STZ group showed an increase in the total cell number. In sum, STZ partially affected female rats, compared to what was previously reported for males. Although AD is more frequent in women, reports about the effect of ICV-STZ in female rats are scarce. Our work highlights the need to deepen into the effects of STZ in the female brain and study possible sex differences.

P-343 Assessing the effect of adipose tissue-derived stem cell-conditioned medium on follicles and stromal cells in cultured ovarian tissue

Abstract Study question Does addition of adipose tissue-derived stem cell-conditioned medium (ASC-CM) enhance follicle survival and development and stromal cell viability in ovarian tissue culture? Summary answer Adding ASC-CM to ovarian tissue in vitro culture (IVC) promotes follicle survival, development, estradiol (E2) production, and stromal cell viability. What is known already Despite extensive efforts to optimize ovarian tissue IVC conditions, outcomes of follicle in vitro development remain suboptimal, mainly showing poor follicle survival and growth. IVC conditions that best resemble the in vivo environment are likely to generate less cellular distress and yield better outcomes. ASCs are known for promoting cell proliferation and survival through secretion of growth factors and cytokines. CM obtained from ASC culture contains these beneficial biomolecules and could potentially be a valuable supplement for ovarian tissue IVC. Study design, size, duration This prospective experimental study used 8 fragments of bovine ovarian cortex, which were cultured for 8 days in two different groups: (i) standard ovarian tissue culture (OT Ctrl D8); and (ii) ovarian tissue cultured with ASC-CM supplementation (OT+CM D8). One fragment per sample served as a non-cultured control (D0). Half of the culture medium was replaced every other day, and medium from days 2 and 8 was stored for further analyses. Participants/materials, setting, methods E2 production was evaluated on days 2 and 8 by ELISA. Follicle classification was established using hematoxylin and eosin (H&E) staining. Follicle and stromal cell apoptosis rates were assessed by TUNEL assays, while growth differentiation factor-9 (GDF-9) labeling served as a follicle quality marker. ASC-CM was analyzed to identify and measure soluble factors like vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and transforming growth factor beta 1 (TGF-β1), associated with protective effects. Main results and the role of chance The OT+CM D8 group showed significantly lower primordial follicle rates (p = 0.03) and higher secondary follicle rates (p = 0.02) than the OT Ctrl D8 group. Concerning apoptosis, the OT+CM D8 group demonstrated significantly lower follicle (p = 0.014) and stromal cell (p = 0.001) rates than the OT Ctrl D8 group. Furthermore, follicles in the OT+CM D8 group exhibited a significant increase (p = 0.002) in GDF-9 expression compared to those in the OT Ctrl D8 group. E2 production was also significantly higher (p = 0.04) after 8 days of culture in the OT+CM D8 group. All studied paracrine factors (VEGF, IL-6 and TGF-b1) were found to be present in the ASC-CM. Limitations, reasons for caution This study focuses only on improving follicle outcomes during the first step of ovarian tissue IVC, where follicles remain in situ within the tissue, and not on subsequent steps. Wider implications of the findings Our study demonstrates that adding ASC-CM to ovarian tissue IVC medium enhances follicle survival, development and E2 secretion, and promotes stromal cell viability. Moreover, our data suggest that ASC-derived factors VEGF, IL-6 and TGF-β1 could be possible paracrine mediators underlying the beneficial effects. Trial registration number not applicable

P-517 The effects of various female infertility factors on endometrial receptivity

Abstract Study question Is there a correlation between different female infertility factors and displaced window of implantation? Summary answer Patients with decreased ovarian reserve and endometriosis have higher risk in experiencing displaced window of implantation. What is known already Numerous studies substantiate that infertility stems from various etiology, including ovulatory dysfunction, male factors, tubal disease, endometriosis, uterine factors...etc. MicroRNAs (miRNAs) play a crucial role in initiating these infertility-related disorders and influencing biological processes like gametogenesis, embryogenesis, ovarian, uterine or endometrial condition. Notably, miRNAs regulate endometrial receptivity, suggesting optimal embryo transfer timing during IVF treatment by identifying the window of implantation (WOI). In recognition of this significant function, MIRA, a molecular testing solution that predicts endometrial receptivity for patients undergoing frozen embryo transfer (FET) by targeting miRNA biomarkers, was developed in hopes of bolstering success rate for personalized embryo transfer. Study design, size, duration This study aims to identify if there is a correlation seen between female infertility factors and the WOI. We enrolled a total of 96 patients who demonstrated successful pregnancy outcomes following personalized embryo transfer. The patients’ personalized embryo transfer timing recommendations were assessed based on their miRNA expression profiles using MIRA. Concurrently, the participants’ underlining infertility diseases were classified according to diagnoses guidelines provided by Taiwan’s Ministry of Health and Welfare. Participants/materials, setting, methods Six categories of female infertility diseases were included: decreased ovarian reserve, tubal, endometriosis, uterine abnormalities (excluding myoma and adenomyosis), polycystic ovary syndrome (PCOS), and myoma factors. Infertile factors such as unexplained, aneuploid oocyte (PGT), male factors, and adenomyosis were excluded due to limited sample size. Statistical analysis of endometrial stages and disease categories involved calculating percentage of the window of implantation (WOI) versus displaced WOI (either pre- or post-receptive) within the six types of diseases. Main results and the role of chance Our result showed that overall, 64.58% of 96 patients have a receptive window of implantation. Compared to this baseline, polycystic ovary syndrome (66.67%) does not elevate the risk of displaced endometrial stages. However, patients with decreased ovarian reserve and endometriosis are more likely to display displaced window of implantation, with average rates of 57.14% and 55.56% WOI respectively. Notably, patients with uterine abnormalities (84.62%) and myoma (87.5%) factors exhibit a lower likelihood of having a displaced WOI. The WOI percentage caused by tubal factors is slightly lower at 76.19%. In conclusion, these preliminary findings establishes that categories of female infertility diseases have discernible effects on patients’ WOI. As patients with certain infertility factors, such as decreased ovarian reserve and endometriosis, demonstrated a greater risk in experiencing displaced WOI, patients that fall under these categories could further benefit from utilizing endometrial receptivity testing to determine the optimal timing for their embryo transfer to potentially improve their reproductive outcomes and facilitate shared decision-making (SDM) in patient’s embryo transfer process. Limitations, reasons for caution While this result offers a preliminary overview for the correlation between endometrial stages and female infertility diseases, we cannot confirm whether these conclusions reached statistical significance due to the limitation of data. Wider implications of the findings Further investigation into the specific reasons for the discernable WOI patterns in different diseases is warranted in larger sample sizes to offer more personalized and robust clinical information supporting IVF treatments. Trial registration number ’not applicable’

Efficacy of omitting initial GnRH in once-used CIDR co-synch protocol for the treatment of anestrus in buffaloes during low breeding season

Progesterone-based hormonal protocols are effective to treat anestrus in buffaloes. However, most of those protocols are costly, thus, requiring a cost-effective protocol that minimizes handling frequencies. This study, therefore, evaluated the effectiveness of the ‘once-used CIDR co-synch protocol’ with or without initial GnRH for treating anestrus in buffaloes during low breeding season. Anestrous buffaloes (n=29) were divided into two Treatment groups. As a part of the ‘once-used CIDR co-synch protocol,’ the first group (T1, n=18) received an initial GnRH on a random day (day 0) while the second group (T2, n=11) did not receive it. On day 0, the once-used CIDR device was inserted intravaginally in both groups, with GnRH administered in T1 but omitted in T2. On day 7, during CIDR removal, each buffalo received PGF2α, and on day 10, GnRH was given, and fixed-timed artificial insemination (FTAI) was done. Ovarian status was assessed on d0 and on 8 to 9-day post-FTAI using transrectal ultrasonography. Plasma progesterone concentrations on 8 to 9-day post-FTAI were determined using ELISA. No significant differences in estrus expression, ovulation and pregnancy rates were observed between two Treatment groups, with overall rates of 93%, 69%, and 34.5%, respectively. However, the overall pregnancy outcome (from FTAI plus natural breeding of buffaloes that returned to estrus) up to 2 months post-FTAI was 48.3%. Parity affected the ovulation among treated buffaloes, with higher (P=0.04) ovulation rate in buffaloes ≤4th parity (79.2%) compared to >4th parity (20%). There were no differences in CL size and plasma progesterone concentrations during 8 to 9-day post-FTAI between two treatment groups. In conclusion, omitting the initial GnRH in ‘once-used CIDR Co-synch protocol’ proved equally effective as the protocol with initial GnRH for the treatment of anestrus in buffaloes during the low breeding season.

The Effect of Ajwa Dates (Phoenix dactylifera L) Consumption to Antral Ovarium Imaging by with Ultrasonography (USG) Transvaginal of Perimenopausal Women

Objectives: Perimenopause is a transition from the reproductive period to the menopause stage with a decrease in the number and function of oocytes. Transvaginal ultrasound is a reliable alternative method for detecting ovarian reserve. Ajwa dates contain phenolic and other flavonoid components which are considered to have the potential to improve the ovarian condition of perimenopausal women which can be assessed by increasing the number of antral follicles as measured by transvaginal ultrasound. This study aims to determine the effect of consuming ajwa dates (Phoenix dactylifera L.) on the appearance of ovarian follicles using transvaginal ultrasound in perimenopausal women. Methods: This research is a quasi-experimental study with a pre-post control design carried out at RSIA Sitti Khadijah I Muhammadiyah Makassar, South Sulawesi, Indonesia, from May to September 2023. This research involved 44 perimenopausal subjects aged 42-48 years who were divided into 2 groups randomly (28 intervention groups, 16 control groups). Subjects in the intervention group consumed seven pieces of ajwa dates (60-70 grams each seven pieces) every morning before having breakfast for eight weeks. The transvaginal USG was performed twice, the first was on day three of the menstrual cycle of each subject and the second was after eight weeks of intervention. A comparison of changes in ovarian follicles was analyzed using the Mann-Whitney U test. Results: This study showed a significant decrease in the number of antral follicles in the intervention group and an increase in the number in the control group (p<0.05). Meanwhile, the size of the antral follicles showed a significant increase in the intervention group and a significant decrease in the size of the antral follicles in the control group (p<0.05). Conclusion: Consuming Ajwa dates as an exogenous antioxidant can affect the number and size of antral follicles, which is an indicator of improving the condition of perimenopausal women's ovaries.International Journal of Human and Health Sciences Supplementary Issue 01: 2024 Page: S66

Brown adipose tissue metabolism in women is dependent on ovarian status.

In rodents, loss of estradiol (E2) reduces brown adipose tissue (BAT) metabolic activity. Whether E2 impacts BAT activity in women is not known. BAT oxidative metabolism was measured in premenopausal (n = 27; 35 ± 9 yr; body mass index = 26.0 ± 5.3 kg/m2) and postmenopausal (n = 25; 51 ± 8 yr; body mass index = 28.0 ± 5.0 kg/m2) women at room temperature and during acute cold exposure using [11C]acetate with positron emission tomography coupled with computed tomograph. BAT glucose uptake was also measured during acute cold exposure using 2-deoxy-2-[18F]fluoro-d-glucose. To isolate the effects of ovarian hormones from biological aging, measurements were repeated in a subset of premenopausal women (n = 8; 40 ± 4 yr; BMI = 28.0 ± 7.2 kg/m2) after 6 mo of gonadotropin-releasing hormone agonist therapy to suppress ovarian hormones. At room temperature, there was no difference in BAT oxidative metabolism between premenopausal (0.56 ± 0.31 min-1) and postmenopausal women (0.63 ± 0.28 min-1). During cold exposure, BAT oxidative metabolism (1.28 ± 0.85 vs. 0.91 ± 0.63 min-1, P = 0.03) and net BAT glucose uptake (84.4 ± 82.5 vs. 29.7 ± 31.4 nmol·g-1·min-1, P < 0.01) were higher in premenopausal than postmenopausal women. In premenopausal women who underwent gonadotropin-releasing hormone agonist, cold-stimulated BAT oxidative metabolism was reduced to a similar level (from 1.36 ± 0.66 min-1 to 0.91 ± 0.41 min-1) to that observed in postmenopausal women (0.91 ± 0.63 min-1). These results provide the first evidence in humans that reproductive hormones are associated with BAT oxidative metabolism and suggest that BAT may be a target to attenuate age-related reduction in energy expenditure and maintain metabolic health in postmenopausal women.NEW & NOTEWORTHY In rodents, loss of estrogen reduces brown adipose tissue (BAT) activity. Whether this is true in humans is not known. We found that BAT oxidative metabolism and glucose uptake were lower in postmenopausal compared to premenopausal women. In premenopausal women who underwent ovarian suppression to reduce circulating estrogen, BAT oxidative metabolism was reduced to postmenopausal levels. Thus the loss of ovarian function in women leads to a reduction in BAT metabolic activity independent of age.

Bilateral ovarian fibroma in postmenopausal woman. Case report and literature review

Ovarian fibromas are rare tumors that mainly occur in perimenopausal and postmenopausal women. Symptoms can vary, and in some cases, they may be associated with ascites and pleural effusion, known as Meigs Syndrome. The article presents the case of a 55-year-old patient with a history of hypertension and bronchial asthma, who visit a gynecologist due to pain in the right iliac fossa of three months duration. Various studies were performed, including ultrasound and abdominal and pelvic CT, which reveal the presence of solid bilateral ovarian tumors. The patient undergoes a total hysterectomy and bilateral salpingo-oophorectomy and despite it’s characteristics, the histopathological study makes the diagnosis of bilateral ovarian fibroma with no signs of malignancy. The clinical presentation of ovarian fibromas is nonspecific, often leading to confusion with malignant ovarian pathology or other benign uterine or ovarian conditions, representing a challenge for preoperative diagnosis.

AI-algorithm training and validation for identification of endometrial CD138+ cells in infertility-associated conditions; polycystic ovary syndrome (PCOS) and recurrent implantation failure (RIF)

BackgroundEndometrial CD138+ plasma cells serve as a diagnostic biomarker for endometrial inflammation, and their elevated occurrence correlates positively with adverse pregnancy outcomes. Infertility-related conditions like polycystic ovary syndrome (PCOS) and recurrent implantation failure (RIF) are closely associated with systemic and local chronic inflammatory status, wherein endometrial CD138+ plasma cell accumulation could also contribute to endometrial pathology. Current methods for quantifying CD138+ cells typically involve laborious and time-consuming microscopic assessments of only a few random areas from a slide. These methods have limitations in accurately representing the entire slide and are susceptible to significant biases arising from intra- and interobserver variations. Implementing artificial intelligence (AI) for CD138+ cell identification could enhance the accuracy, reproducibility, and reliability of analysis. MethodsHere, an AI algorithm was developed to identify CD138+ plasma cells within endometrial tissue. The AI model comprised two layers of convolutional neural networks (CNNs). CNN1 was trained to segment epithelium and stroma across 28,363 mm2 (2.56 mm2 of epithelium and 24.87 mm2 of stroma), while CNN2 was trained to distinguish stromal cells based on CD138 staining, encompassing 7345 cells in the object layers (6942 CD138− cells and 403 CD138+ cells). The training and performance of the AI model were validated by three experienced pathologists. We collected 193 endometrial tissues from healthy controls (n = 73), women with PCOS (n = 91), and RIF patients (n = 29) and compared the CD138+ cell percentages based on cycle phases, ovulation status, and endometrial receptivity utilizing the AI model. ResultsThe AI algorithm consistently and reliably distinguished CD138− and CD138+ cells, with total error rates of 6.32% and 3.23%, respectively. During the training validation, there was a complete agreement between the decisions made by the pathologists and the AI algorithm, while the performance validation demonstrated excellent accuracy between the AI and human evaluation methods (intraclass correlation; 0.76, 95% confidence intervals; 0.36–0.93, p = 0.002) and a positive correlation (Spearman's rank correlation coefficient: 0.79, p < 0.01). In the AI analysis, the AI model revealed higher CD138+ cell percentages in the proliferative phase (PE) endometrium compared to the secretory phase or anovulatory PCOS endometrium, irrespective of PCOS diagnosis. Interestingly, CD138+ percentages differed according to PCOS phenotype in the PE (p = 0.03). On the other hand, the receptivity status had no impact on the cell percentages in RIF samples. ConclusionOur findings emphasize the potential and accuracy of the AI algorithm in detecting endometrial CD138+ plasma cells, offering distinct advantages over manual inspection, such as rapid analysis of whole slide images, reduction of intra- and interobserver variations, sparing the valuable time of trained specialists, and consistent productivity. This supports the application of AI technology to help clinical decision-making, for example, in understanding endometrial cycle phase-related dynamics, as well as different reproductive disorders.

Resumption of ovulation in anovulatory women with PCOS and obesity is associated with reduction of 11β-hydroxyandrostenedione concentrations.

Is resumption of ovulation after a 6-month lifestyle intervention in women with PCOS and obesity associated with differential changes in endocrine and metabolic parameters (weight, insulin resistance, anti-Müllerian hormone (AMH), and androgens) compared to women with PCOS who remained anovulatory? Resumption of ovulation after a 6-month lifestyle intervention in women with PCOS and obesity is associated with changes in serum 11β-hydroxyandrostenedione (11OHA4) concentrations. Lifestyle interventions have been shown to reduce clinical and biochemical hyperandrogenism in women with PCOS. Weight loss of 5-10% may reverse anovulatory status, thereby increasing natural conception rates. However, the mechanisms underlying why some women with PCOS remain anovulatory and others resume ovulation after weight loss are unclear. Reproductive characteristics at baseline and a greater degree of change in endocrine and metabolic features with lifestyle intervention may be crucial for ovulatory response. We used data and samples originating from an earlier randomized controlled trial (RCT), which examined the efficacy of a 6-month lifestyle intervention prior to infertility treatment compared to prompt infertility treatment on live birth rate in women with obesity. A total of 577 women with obesity (BMI > 29 kg/m2) were randomized between 2009 and 2012. Anovulatory women with PCOS who were allocated to the intervention arm of the original RCT (n = 95) were included in the current analysis. We defined women as having resumed ovulation (RO+) based on the following criteria: spontaneous pregnancy; or assignment to expectant management; or IUI in natural cycles as the treatment strategy after lifestyle intervention. Steroid hormones were measured using liquid chromatography tandem mass spectrometry. Generalized estimating equations with adjustment for baseline measures and interaction between group and time was used to examine differences in changes of endocrine and metabolic parameters between RO+ (n = 34) and persistently anovulatory women (RO-, n = 61) at 3 and 6 months after intervention. At baseline, the mean ± SD age was 27.5 ± 3.6 years in the RO+ group and 27.9 ± 4.1 years in the RO- group (P = 0.65), and the mean ± SD weights were 101.2 ± 9.5 kg and 105.0 ± 14.6 kg, respectively (P = 0.13). Baseline AMH concentrations showed significant differences between RO+ and RO- women (median and interquartile range [IQR] 4.7 [3.2; 8.3] versus 7.2 [5.3; 10.8] ng/ml, respectively). Baseline androgen concentrations did not differ between the two groups. During and after lifestyle intervention, both groups showed weight loss; changes in 11OHA4 were significantly different between the RO+ and RO groups (P-value for interaction = 0.03). There was a similar trend for SHBG (interaction P-value = 0.07), and DHEA-S (interaction P-value = 0.06), with the most pronounced differences observed in the first 3 months. Other parameters, such as AMH and FAI, decreased over time but with no difference between the groups. No high-resolution transvaginal ultrasonography was used to confirm ovulatory status at the end of the lifestyle program. The small sample size may limit the robustness of the results. Reduction of androgen concentrations during and after lifestyle intervention is associated with recovery of ovulatory cycles. If our results are confirmed in other studies, androgen concentrations could be monitored during lifestyle intervention to provide individualized recommendations on the timing of resumption of ovulation in anovulatory women with PCOS and obesity. The study was supported by a grant from ZonMw, the Dutch Organization for Health Research and Development (50-50110-96-518). The Department of Obstetrics and Gynecology of the UMCG received an unrestricted educational grant from Ferring Pharmaceuticals BV, The Netherlands. A.H. reports consultancy for the development and implementation of a lifestyle App MyFertiCoach developed by Ferring Pharmaceutical Company. All other authors have no conflicts to declare. The LIFEstyle RCT was registered at the Dutch trial registry (NTR 1530).

Dairy village: the role of veterinary services in unlocking dairy industry potential through assisted reproductive technologies

The “Dairy Village” concept was designed to overcome the constraints of estrus synchronization and AI particularly associated with eletion of the breeding animals and subsequent management of estrous cows in Ethiopia. All necessary inputs, including sexed semen, were acquired before the fieldwork. The knowledge and skill gaps were identified and addressed with the active involvement of veterinarians and AI technicians who underwent specialized training to ensure proper physical and gynecological examinations during the selection of cows and heifers. Accordingly, a refresher training was delivered on topics of reproductive anatomy and physiology, assessment of ovarian status, semen handling, and insemination at the optimal time to establish pregnancy. Besides, awareness creation seminars were held every year on topics of benefits of reproductive technologies and management of breeding cows to 99 smallholder dairy farmers. Only 578 cows/heifers were found fit for breeding out of the 1262 cows/heifers that were brought for evaluation; including 81 animals that spontaneously showed estrus. A total of 497 cows/heifers were synchronized using a single IM injection of Lutalyse. Animals responding to the treatment (404 cow/heifers) and those spontaneously in estrus (81cows/ heifers) were inseminated using sex- sorted HF semen 18-24hrs into the standing estrus. Pregnancy rate was 80% (388/485) and 287 calves were born (96% female). Results of this project indicate that rigorous gynecological examination during the selection of cows prior to estrus synchronization and judicious application of hormonal treatments can yield improve the success of ES/AI projects under field conditions. Furthermore, the dairy village approach to enhanced milk production through efficient use of sex-sorted semen has generated replacement heifers from superior breed that will substantially improve the livelihoods of village farmers.

Abstract A111: Development and analytical validation of a multiplexed targeted mass spectrometry assay to detect ovarian cancer protein biomarkers in sera

Abstract A111: Development and analytical validation of a multiplexed targeted mass spectrometry assay to detect ovarian cancer protein biomarkers in sera

ASSOCIATION OF LOW BMI AND ANOVULATION WITH PRIMARY INFERTILITY IN WOMEN OF REPRODUCTIVE AGE

Infertility, defined as the inability to conceive after one year of regular unprotected intercourse, affects millions of couples worldwide, with prevalence rates varying across developed and developing nations. The main objective of the study is to find the association between low BMI and anovulation with primary infertility in women of reproductive age. This cross-sectional study was conducted in Khyber Teaching Hospital Peshawar from January 2023 to December 2023. A total of 340 female participants were added using convenient sampling methods. Data collection consisted of comprehensive clinical evaluations, including anthropometric measurements to assess BMI, hormonal assays to determine ovulatory status and detailed reproductive histories. BMI was calculated as weight in kilograms divided by height in meters squared (kg/m^2), with participants categorized into low BMI (BMI &lt; 18.5 kg/m^2) and normal BMI (BMI ≥ 18.5 kg/m^2) groups. Data were collected from 340 female participants. The mean age was 28.5± 4.2 years for those with low BMI and 30.1± 3.8 years for those with normal BMI. Among underweight individuals, 50 were classified as infertile and 20 as fertile, totaling 70 participants. Of normal-weight individuals, 40 were infertile, and 120 were fertile, totaling 160 participants. In the overweight group, 30 were barren, and 50 were fertile, totaling 80 participants. Low BMI showed a significantly increased adjusted odds ratio (AOR) of 3.00 (95% CI: 1.80 - 5.00, p &lt; 0.001) for infertility. Age, socioeconomic status, smoking, and alcohol consumption did not show significant associations with infertility. It is concluded that low BMI is significantly associated with an increased risk of anovulation and primary infertility in women of reproductive age, even after accounting for potential confounders.

Ovarian tissue cryopreservation for fertility preservation before hematopoietic stem cell transplantation in patients with sickle cell disease: safety, ovarian function follow-up, and results of ovarian tissue transplantation.

To describe the experience of performing ovarian tissue cryopreservation (OTC) before hematopoietic stem cell transplantation (HSCT), among girls/women with severe sickle cell disease (SCD)(SS or S/β0-thalassemia) who are, besides the usual surgical risk, at risk of SCD-related complications during the fertility preservation procedure for improving their counseling and management. This retrospective study included 75 patients (girls/women) with SCD who have had OTC before myeloablative conditioning regimen (MAC) for HSCT. Characteristics of patients and data on OTC, ovarian status follow-up, and results of ovarian tissue transplantation (OTT) were collected in medical records. At OTC, the median (IQR 25-75; range) age of the patients was 9.6 (6.9-14.1; 3.6-28.3) years, 56/75 were prepubertal, and no SCD or surgery-related complications occurred. The median follow-up post-HSCT was > 9years. At the last follow-up, among prepubertal patients at HSCT, 26/56 were ≥ 15years old and presented with a premature ovarian insufficiency (POI), except 2, including the patient who had received an OTT to induce puberty. Eight were 13-15years old and presented for POI. The remaining 22 patients were under 13. Among the 19 patients who were menarche at HSCT, 2 died 6months post-HSCT and we do not have ovarian function follow-up for the other 2 patients. All the remaining patients (n = 15) had POI. Five patients had OTT. All had a return of ovarian function. One patient gave birth to a healthy baby. OTC is a safe fertility preservation technique and could be offered before MAC independent of the patient's age.

Analysis of production efficiency of cloned transgenic Yucatan miniature pigs according to recipient breeds with embryo transfer conditions

The purpose of this study was to compare the efficiency of the production of cloned transgenic Yucatan miniature pigs (YMPs) using two recipient breeds, i.e., YMPs and domestic pigs (DPs), under various embryo transfer conditions. We initially assessed the in vitro developmental competence of embryos obtained via somatic cell nuclear transfer (SCNT) from three different transgenic donor cells. No difference was observed among the three groups regarding developmental competence. Furthermore, the cloning efficiency remained consistent among the three groups after the transfer of the SCNT embryos to each surrogate mother. Subsequently, to compare the efficiency of the production of cloned transgenic YMPs between the two recipient breeds using varying parameters, including ovulation status (preovulation and postovulation), duration of in vitro culture (IVC) (incubated within 24 h and 24–48 h), and the number of transferred SCNT embryos (less than and more than 300), we assessed the pregnancy rates, delivery rates, mean offspring counts, and cloning efficiency. Regarding the ovulation status, YMPs exhibited higher pregnancy rates, delivery rates, and cloning efficiency compared with DPs in both statuses. Moreover, the pregnancy rates, delivery rates, and cloning efficiency were affected by the ovulation status in DPs, but not in YMPs. The comparison of IVC duration between groups revealed that YMPs had higher pregnancy rates vs. DPs in both conditions. SCNT embryos cultured for 24–48 h in YMPs yielded higher delivery rates and cloning efficiency compared with those cultured for less than 24 h in DPs. Finally, the analysis based on the number of transferred SCNT embryos showed that both the pregnancy and delivery rates were higher in YMPs vs. DPs. However, the highest average number of offspring was obtained when more than 300 SCNT embryos were transferred into DPs, whereas the cloning efficiency was higher in YMPs vs. DPs. Our results suggest that YMPs are more suitable recipients than are DPs under various conditions for the production of cloned transgenic YMPs.

Multi-Omics Reveals the Role of Osteopontin/Secreted Phosphoprotein 1 in Regulating Ovarian Aging.

Secreted phosphoprotein 1 (SPP1), also known as osteopontin (OPN), is located on chromosome 4q22.1. This multifunctional secreted acidic glycoprotein is expressed intracellularly and extracellularly in various tissues, where it interacts with regulatory proteins and pro-inflammatory immune chemokines, contributing to the pathogenesis of multiple diseases. Nevertheless, the intricate genetic connections between SPP1 and ovarian aging remain largely unexplored. This study aims to bridge this knowledge gap by delving into ovarian aging and its associations with SPP1 using multi-omics data analysis. Our findings indicate that SPP1 is a potential gene related to ovarian aging. To comprehend the role of SPP1, we conducted spatial transcriptomic analyses on young and aged female mouse ovaries, revealing a significant decline in SPP1 expression in the aging group compared to the young group. Similarly, a significantly low level of SPP1 was found in the 73-year-old sample. Additionally, in-depth single-cell RNA-sequencing analysis identified associations between SPP1 and ITGAV, ITGB1, CD44, MMP3, and FN1. Notably, co-expression analysis highlighted a strong correlation between SPP1 and ITGB1. In summary, this study pioneers the identification of SPP1 as a gene implicated in ovarian aging. Further research into the role of SPP1 has the potential to advance precision medicine and improve treatment strategies for ovarian aging-related conditions.