Antral Follicle Diameter Research Articles

In vivo study: black cumin extract (Nigella sativa) on antral follicle diameter and granulosa cell layer thickness as an effort to prevent infertility

Background: Infertility is a health problem that affects approximately 10-15% of couples trying to achieve pregnancy worldwide. Various studies have shown that Nigella sativa (N. Sativa) has many health benefits, including reproductive health. However, studies examining the effectiveness of N. Sativa in preventing folliculogenesis disorders have never been reported.Purpose: This study aims to determine the benefits of administering N. Sativa in preventing folliculogenesis disorders as measured by the diameter of the antral follicles and the thickness of the granulosa cell layer.Methods: This research design uses an experiment with a randomized posttest-only controlled group design approach. There were 27 experimental animals (mice) divided into three groups, namely the negative control group (9 animals) and the positive control group (9 animals), as well as the treatment group (9 animals). The positive control group was exposed to two cigarettes of cigarette smoke per day. The treatment group was exposed to cigarette smoke and N. Sativa 300 mg/KgBW/day for 28 days. After that, the mice were sacrificed to take their ovaries. Then, the ovarian organ samples were stained with Hematoxylin-eosin and examined for the diameter of the antral follicles and the thickness of the granulosa cell layer in the antral follicles with photo slides at 400x magnification in 5 fields of view, statistical analysis using the Independent T-test.Results: There were significant differences in antral follicle diameter and granulosa cell thickness in mice exposed to cigarette smoke and N. Sativa. Administration of N. Sativa extract can increase granulosa cell thickness through estrogenic function. Administration of N. Sativa extract showed estrogenic activity by increasing serum estradiol levels. N. Sativa heterotrophic activity cannot be separated from the flavonoid and phenolic compounds contained in N. Sativa, which have been proven to have high estrogenic activity.Conclusion: N. Sativa extract is efficacious in improving the diameter of antral follicles and the thickness of granulosa cells caused by exposure to cigarettes.

Associations among the largest follicle, preovulatory estradiol concentrations, and predominant vaginal epithelial cells at the completion of hormonal ovarian stimulation for fixed-time artificial insemination in goats.

The objective of the present study was to investigate the association among the largest follicle (LF), preovulatory estradiol (E2), and predominant vaginal epithelial cell at the completion of hormonal ovarian stimulation for fixed-time artificial insemination (FTAI) in goats. Thirty-seven crossbred Boer does received gonadotropin-releasing hormone (GnRH) and intravaginal progesterone (P4)-releasing devices (day 0). On day 5, P4 devices were removed and does received prostaglandin F2α and equine chorionic gonadotrophin. On day 7, does received GnRH, and FTAI was undertaken. On day 7, does were divided into three groups, i.e. small-sized (3-3.9 mm; n = 5), medium-sized (4-4.9 mm; n = 8), and large-sized (≥5 mm; n = 24) according to the diameter of the ovarian LF; follicular characteristics (number and diameter) were identified, and blood samples and vaginal smears were collected. The average diameters of total antral follicles and LF and the percentage of superficial cell were greatest in large-sized LF does (p < .01). The average diameters of total antral follicle (r = .68) and LF (r = .71), number of preovulatory follicle (r = .58), and plasma E2 concentrations (r = .61) were positively correlated with the percentage of superficial cells (p < .01). The likelihood of a pregnancy outcome after the FTAI increased by 13.71 times in does with a greater average diameter of antral follicle, 14.18 times with emergence of a large preovulatory follicle, and 36.83 times with a higher percentage of vaginal superficial cells (p < .01). It was concluded that there is a relationship between the cell types of the vaginal epithelium, the diameters of the largest ovarian follicles, and the concentration of E2 in goats subjected to FTAI protocols.

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.

Do we overlook predictive factors in Poseidon 1 patients? A retrospective analysis co-evaluating antral follicle counts & diameters

BackgroundAn unexpected impaired ovarian response pertains to an insufficient reaction to controlled ovarian hyperstimulation. This deficient reaction is identified by a reduced count of mature follicles and retrieved oocytes during an IVF cycle, potentially diminishing the likelihood of a successful pregnancy. This research seeks to examine whether the characteristics of antral follicles can serve as predictive indicators for the unexpected impaired ovarian response to controlled ovarian stimulation (COS).MethodsThis retrospective cohort study was conducted at a tertiary university hospital. The electronic database of the ART (assisted reproductive technologies) center was screened between the years 2012–2022. Infertile women under 35 years, with normal ovarian reserve [anti-Müllerian hormone (AMH) > 1.2 ng/ml, antral follicle count (AFC) > 5] who underwent their first controlled ovarian stimulation (COS) cycle were selected. Women with < 9 oocytes retrieved (group 1 of the Poseidon classification) constituted the group A, whereas those with ≥ 9 oocytes severed as control (normo-responders) one (group B). Demographic, anthropometric and hormonal variables together with COS parameters of the two groups were compared.ResultsThe number of patients with < 9 oocytes (group A) was 404, and those with ≥ 9 oocytes were 602 (group B). The mean age of the group A was significantly higher (30.1 + 2.9 vs. 29.4 + 2.9, p = 0.01). Group A displayed lower AMH and AFC [with interquartile ranges (IQR); AMH 1.6 ng/ml (1-2.6) vs. 3.5 ng/ml (2.2–5.4) p < 0.01, AFC 8 (6–12) vs. 12 (9–17), p < 0.01]. The number of small antral follicles (2–5 mm) of the group A was significantly lower [6 (4–8) vs. 8 (6–12) p < 0.01), while the larger follicles (5–10 mm) remained similar [3 (1–5) vs. 3(1–6) p = 0.3] between the groups.ConclusionThe propensity of low ovarian reserve and higher age are the main risk factors for the impaired ovarian response. The proportion of the small antral follicles may be a predictive factor for ovarian response to prevent unexpected poor results.

Naproxen administration affects murine late folliculogenesis, reduces granulosa cell proliferation and the number of ovulated oocytes

Naproxen reduces the production of prostaglandins via inhibition of the cyclooxygenase. Studies have shown that its administration in women can be related to failed ovulation. Therefore, preclinical investigations must be performed in order to investigate its effects in experimental models. Thus, the aim of this study was to evaluate the effects of naproxen on murine folliculogenesis, ovulation, and female fertility. Female C57BL/6 mice (n = 128 - 6 weeks old) were divided into Control, low (10 mg/kg), and high naproxen (50 mg/kg) groups, who were treated for 8 days and directed to morphofunctional analyses. Follicular quantification showed a reduced percentage of antral follicles in naproxen-treated animals. These treated animals also showed smaller oocytes included in secondary and antral follicles, and the diameter of secondary and antral follicles was also reduced. A reduction in the percentage of Ki67-positive granulosa cells was observed in treated animals that also showed down-regulation of Igf1r compared to control. After an ovarian stimulation protocol, naproxen-treated animals showed a reduction in the percentage of secondary and antral follicles, a reduced number of ovulated oocytes and, corpora lutea, and an increased number of failed ovulations. Finally, naproxen-treated animals also showed a reduction in mating index and pregnancy rate. Our findings suggested that, in mice, naproxen administration (eight days treatment) negatively affects molecular and morphological aspects related to late folliculogenesis, ovulation, and fertility.

The Comparative Effect of Citrullus colocynthis Hydro-Alcoholic Extract and Metformin on Morphometric Ovarian Follicles Disorders in Estradilol valerate-Iinduced Polycystic Ovary Syndrome Rats

Background: Polycystic ovary syndrome (PCOS) is an endocrine disorder that affects 6-10% of women in reproductive age. The medicinal values of Citrullus colocynthis extract (CCT) stems from its anti-oxidative and anti-inflammatory effects. This study evaluated the comparative effect of CCT and metformin on morphometric ovarian disorders in PCOS rats. Materials and Methods: Fifty female Wistar rats having 2-3 consecutive estrous cycles during two weeks were divided into five groups (n=10 each group). The rats in the control group only received sesame oil as estradiol valerate solvent, whereas the rats in the sham group were injected intramuscularly with 4 mg/rat estradiol valerate-induced PCOS. Following PCOS induction, the rats in the metformin group received 50 mg/kg metformin orally for three weeks. Rats in the Extract group were treated with 50 mg/kg plant extract for 21 days following the induction with PCOS. Additionally, the rats in metformin+ extract group were treated with the combination of 50 mg metformin plus plant extract for three weeks. The ovaries were removed and were fixed for tissue processing. The slices were stained by hematoxylin-eosin after clearing and hydration. Follicular count and morphometric changes were evaluated in primordial, primary, pre-antral and antral follicles. Results: The mean diameter of primordial follicle was similar in all groups, but mean diameter of primary follicle decreased in the sham group (145.50± 49.26 µm) compared to metformin (278.76± 156.64 µm), extract (311.70± 147.74 µm) and metformin+ extract groups (265.35± 88.16 µm). The diameter of pre-antral and antral follicles in the sham group were significantly larger than those in the control group, but were not significantly different in all other groups including metformin, extract, and metformin+ extract in comparison with control group (P&lt; 0.05). Conclusion: In this study, the data has demonstrated that CCT like metformin could improve follicular morphometric disorders in PCOS rats. [GMJ.2018;7:e1045] Â

FRI422 The Fates Of Ovarian Dominant Follicles In The First Two Gynecologic Years: An Investigation Of Morphologic And Endocrinologic Features

Abstract Disclosure: M. Calvert: None. S. Molsberry: None. S. Denslow: None. H. Vanden Brink: None. M.E. Lujan: None. N. Shaw: None. Introduction: Irregular menses are common during the early, post-menarchal years owing to the immaturity of the hypothalamic-pituitary-ovarian axis. Few studies have attempted to delineate the factors that contribute to cycle irregularity during this time. In the current study, we aimed to characterize the fate of ovarian dominant follicles in a longitudinal cohort of healthy, early post-menarchal girls. Methods: Participants contributed daily urine samples for reproductive hormone profiling (Cr-corrected LH, estrone conjugates [E1C], and pregnanediol-3-glucuronide [PdG]), and underwent periodic transabdominal ultrasound scans and body composition analysis. The current analysis includes cycles: 1) with an LH surge and ultrasounds performed 1-9 days before and/or &amp;lt;11 days after the surge, or 2) without an LH surge but with ultrasound documentation of a dominant follicle. LH surge requirements were &amp;gt; 40 mIU/mg Cr with concentrations &amp;gt; mean+3sd of the preceding 5 days. Ultrasounds were evaluated offline by two investigators using a standardized rubric for ovarian structures. Results: 26 participants contributed 74 ultrasound scans, spanning 45 cycles. Participants were 12.6 (range 9.8-14.7) years-old, 2.7 (range 0.3-9.6) months post-menarche, predominantly non-Hispanic White (65.4% vs. 26.9% Black, 3.8% Asian, 3.8% Multiracial), and of normal weight (76.9% with BMI &amp;lt;85th percentile). In 41/45 cycles, there was morphological (antral follicle diameter &amp;gt; 10 mm observed ultrasonographically, n=22) and/or endocrinological (LH surge, n=38) evidence for the presence of a dominant follicle. In 4/45 cycles, a dominant follicle was observed but peak LH did not meet LH surge criteria. In 17/41 cycles, dominant follicle fate could not be determined for technical reasons. In the remaining 24/41 cycles, dominant follicles fates post LH-surge were: corpus luteum (CL, 15/24, 71%), CL or luteinized unruptured follicle [LUF] (2/24, 8%), persistent dominant follicle (3/24, 13%), or no structure observed, implying follicular regression/collapse (4/24, 17%). In logistic GEE models, there was no association between peak E1C or LH, follicular phase length, gynecologic age, or free testosterone with odds of having a CL versus any other dominant follicle outcome. There were marginally significant associations between peak PdG (OR: 1.0003, 95% CI: 0.9999, 1.00008) and total body fat percent (OR: 0.86, 95% CI: 0.74, 1.01) with odds of having a CL versus any other dominant follicle outcome. Conclusion: In the first two gynecologic years, dominant follicles develop in most cycles and frequently ovulate. A variety of mechanisms, however, contribute to anovulation, including an absent LH surge, the formation of LUFs, and follicle regression. Whether degree of adiposity is related to ovulatory success of a dominant follicle early in gynecological life will be pursued in future research. Presentation: Friday, June 16, 2023

Zinc and α-tocopherol protect the antral follicles and endogenous antioxidants of female albino rats (Rattus norvegicus) against lead toxicity

BackgroundLead impairs female reproductive health because it can induce oxidative stress. Zinc as an antioxidant produces an enzyme system that helps neutralize free radicals. α-Tocopherol has an antagonistic effect that reduces oxidative stress. This study aimed to demonstrate the effects of zinc (Zn) and α-tocopherol on the ovarian endogenous antioxidants and antral follicles of albino rats (Rattus norvegicus) exposed to lead acetate (Pb(C2H3O2)2). MethodsTwenty-five female Wistar rats were divided into five groups, namely groups K (control), P0, P1, P2, and P3. Following exposure and treatment for 21 days with different combinations, the albino rats were necropsied, and their ovaries were removed for subsequent histopathological preparations and endogenous antioxidant analysis. Observations were made on the ovary, including an antral follicle count and diameter calculations. Analysis of the superoxide dismutase (SOD) levels (560 nm wavelength) and malondialdehyde MDA-TBA (532 nm wavelength) were performed by a spectrophotometer. The data were analyzed using a one-way ANOVA and least significant difference (LSD) test with the SPSS V24 software. ResultsThe highest SOD enzyme expression in the albino rat ovaries was in P0 (17.23 ± 5.34), and the lowest was in P3 (4.21 ± 0.76). The lowest MDA level was observed in the control group (K) and P3 compared to the other groups. The highest average antral follicle count and diameter were found in the albino rats exposed to 1.5 mg/kg BW lead acetate, and treated with 0.54 mg/kg BW zinc sulfate and 100 mg/kg BW α-tocopherol (group P3) compared to the other groups. The mechanisms of action of zinc and α-tocopherol work synergistically to decrease free radicals and ovarian damage. ConclusionThe results showed that a combination of 0.54 mg/kg BW zinc (Zn) and 100 mg/kg BW α-tocopherol can maintain the number and diameter of the antral follicles and reduce ovarian SOD expression and MDA levels in albino rats exposed to lead acetate.

Age-related changes in circulating INSL3 concentrations and their associations with ovarian conditions in Japanese Black beef cattle

Information on circulating levels of insulin-like peptide 3 (INSL3) in female domesticated animals is limited, as their concentrations are significantly lower than in males. The objectives of the present study were to 1) develop a sandwich time-resolved fluorescence immunoassay (TRFIA) with higher detectability to measure blood INSL3 concentrations in female cattle, 2) determine INSL3 concentrations in female cattle among age groups and reproductive conditions, and 3) explore associations between INSL3 levels and ultrasonographic ovarian measurements. Blood was collected repeatedly from Japanese Black beef female calves (n = 12; 0－8 mo), heifers (n = 10; 10－26 mo), and cows (n = 20; 27－200 mo). Blood was taken from the cows (n = 13) at follicular, post-ovulatory, and luteal phases, and from cows with follicular cysts (n = 12). Ultrasonography of ovaries was conducted in the calves (n = 12) and the cows without ovarian diseases (n = 9). The ovarian area, as well as the number and diameters of antral follicles ≥ 2 mm, were determined in each ovary. The proposed method detected a difference in plasma INSL3 between calves (0.01 ng/mL) and heifers (0.18 ng/mL). However, the conventional assay showed similar levels for calves and heifers (1.82 vs 2.07 ng/mL). Plasma INSL3 and testosterone concentrations increased from calves to heifers (P < 0.0001), but only INSL3 rose from heifers to cows (P < 0.0001). INSL3 and testosterone concentrations did not change across the estrus cycle in cows, and the levels of both hormones in follicular cystic cows did not differ from those in the follicular phase. Ovarian area, maximal and average follicular diameters, and total volume of all follicles per animal were higher in cows than calves (P < 0.001). Plasma INSL3 concentrations correlated positively with the total volumes of all follicles in calves (P < 0.05) and cows (P < 0.05), whereas testosterone concentrations did not correlate with ovarian follicular measurements. In conclusion, plasma INSL3 concentrations measured by the proposed sandwich TRFIA showed a clear increase from female calves to cows in beef cattle. These results suggest that circulating levels of INSL3, but not of testosterone, are associated with the total volume of all antral follicles in both ovaries per animal in female cattle.

Role of gonadotropin-releasing hormone-II and its receptor in swine reproduction.

The pig represents the only livestock mammal capable of producing a functional protein for the second mammalian form of gonadotropin-releasing hormone (GnRH-II) and its receptor (GnRHR-II). To examine the role of GnRH-II and its receptor in pig reproduction, we produced a unique swine line with ubiquitous knockdown of endogenous GnRHR-II levels (GnRHR-II knockdown[KD]), which is largely the focus of this review. In mature GnRHR-II KD males, circulating testosterone concentrations were 82% lower than littermate control boars, despite similar luteinizing hormone (LH) levels. In addition, nine other gonadal steroids were reduced in the serum of GnRHR-II KD boars, whereas adrenal steroids (except 11-deoxycortisol) did not differ between lines. Interestingly, testes from GnRHR-II KD males had fewer, hypertrophic Leydig cells and fewer, enlarged seminiferous tubules than control testes. As expected, downstream reproductive traits such as androgen-dependent organ weights and semen characteristics were also significantly reduced in GnRHR-II KD versus control boars. Next, we explored the importance of this novel ligand/receptor complex in female reproduction. Transgenic gilts had fewer, but heavier, corpora lutea with smaller luteal cells than littermate control females. Although the number of antral follicles were similar between lines, the diameter of antral follicles tended to be larger in GnRHR-II KD females. In regard to steroidogenesis, circulating concentrations of progesterone and 17β-estradiol were lower in transgenic compared to control gilts, even though serum levels of follicle-stimulating hormone and LH were similar. Thus, GnRH-II and GnRHR-II represent a potential avenue to enhance fertility and promote the profitability of pork producers.

Evaluation of correlation between antral follicle diameters with Follicular Output Rate (FORT) in women under controlled ovarian hyperstimulation for assisted reproductive techniques

Abstract Background This study aimed to determine the association of diametrical antral follicles with the ovarian response by Follicular Output Rate (FORT) ratio in 100 females undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). This study selected 100 women undergoing controlled ovarian hyperstimulation (COH). The number of antral follicles (diameter of 3–10 mm) was calculated with two-dimensional vaginal sonography on days 1–3 of the cycle. Then, on a triggering day with human chorionic gonadotropin (HCG), the number of follicles (with a diameter of 16–22 mm) and the ratio of FORT were determined. The correlation among FORT with age, antral follicle count (AFC), AFC ≤ 5, AFC &gt; 5, number of preovulatory follicles (16–20 mm), number of metaphase II (MII) oocytes, body mass index (BMI), infertility period, and anti-Mullerian hormone (AMH) was assessed. Results There was a significant correlation between FORT and total AFC, AFC &gt; 5, number of preovulatory follicles (16–20 mm), and number of MII oocytes retrieved. There is no significant relationship between FORT and examined two variables (AMH and AFC ≤ 5). Multiple linear regression analysis showed no significant relationship between FORT and examined two variables (AMH and AFC &gt; 5). There was a significant correlation between MII oocytes retrieved and age, total AFC, AFC ≤ 5, AFC &gt; 5, number of preovulatory follicles (16–20 mm), and AMH. A significant positive relationship existed between MII oocytes retrieved and examined two variables (AFC ≤ 5 and AFC &gt; 5). Conclusion There was not much difference in the correlation between the AFC ≤ 5 and AFC &gt; 5, and both positively correlated with the number of MII oocytes retrieved.

What should be the strategy in case of a big follicle at the start of the cycle? Shall we start the stimulation or postpone it to the next cycle?

This study facilitates decision-making when an antral follicle diameter >15 mm is detected at the beginning of the menstrual cycle in poor responder (POR) patients. Eighty-three POR patients with at least one leading follicle with a diameter of 15 to 24 mm on the 2nd-4th days of the menstrual cycle were assessed. The mean age of females was 40.1±4.8 (26-45), and the mean partners' age was 42.1±7.8 (26-65). Fifty-one (61.4%) women underwent an oocyte pick-up procedure 36 h after the first ultrasonographic examination on the 2nd-4th days of the menstrual cycle. Gonadotrophin stimulation was initiated in 32 (38.6%) patients. Among women in whom oocyte retrieval was performed, an oocyte was obtained in 49 (59.75%) patients. In 13 of 49 patients (26.5%), no mature oocytes were obtained. Fertilized 2pn embryos were obtained in 18 of 33 patients (54.5%). Among the fertilized embryos, 12 were good, six were moderate, and two were of poor quality. Following the frozen embryo transfer procedure, one of the two patients experienced a clinical pregnancy. Patients with POR are still difficult to manage both clinically and therapeutically. Since every oocyte is valuable and important, patients should be carefully followed up. Our research will be directed by the need to rule out a physiological ovarian cyst when large antral follicles appear at the beginning of the cycle. The clinician should give them a chance.

Diversity of Mitochondrial DNA Haplogroups and Their Association with Bovine Antral Follicle Count.

Simple SummaryChinese Holstein cows, crossbred cattle in China, have abundant genetic diversity due to the fact that Holstein cows were crossbred with various local breeds. Therefore, to reveal their genetic diversity, we sequenced the complete mitochondrial DNA (mtDNA) D-loop region of 501 Chinese Holstein cows and found abundant genetic resources (including 219 haplotypes, polymorphisms of 260 SNPs, and 32 indels) and two haplogroups in the study population. Meanwhile, the two haplogroups had different maternal origins and corresponding distinguished mutations. Furthermore, the association between haplogroups and antral follicle count was found, and the mtDNA haplogroup HG1 with Bos indicus maternal origin had more antral follicles (diameter ≥ 8 mm), implying that these individuals of HG1 had higher reproductive potential. Finally, these distinguished mutations between two haplogroups could be considered as important genetic markers for cattle breeding.Maternal origins based on the bovine mitochondrial D-loop region are proven to have two main origins: Bos taurus and Bos indicus. To examine the association between the maternal origins of bovine and reproductive traits, the complete mitochondrial D-loop region sequences from 501 Chinese Holstein cows and 94 individuals of other breeds were analyzed. Based on the results obtained from the haplotype analysis, 260 SNPs (single nucleotide polymorphism), 32 indels (insertion/deletion), and 219 haplotypes were identified. Moreover, the nucleotide diversity (π) and haplotype diversity (Hd) were 0.024 ± 0.001 and 0.9794 ± 0.003, respectively, indicating the abundance of genetic resources in Chinese Holstein cows. The results of the median-joining network analysis showed two haplogroups (HG, including HG1 and HG2) that diverged in genetic distance. Furthermore, the two haplogroups were significantly (p < 0.05) correlated with the antral follicle (diameter ≥ 8 mm) count, and HG1 individuals had more antral follicles than HG2 individuals, suggesting that these different genetic variants between HG1 and HG2 correlate with reproductive traits. The construction of a neighbor-joining phylogenetic tree and principal component analysis also revealed two main clades (HG1 and HG2) with different maternal origins: Bos indicus and Bos taurus, respectively. Therefore, HG1 originating from the maternal ancestors of Bos indicus may have a greater reproductive performance, and potential genetic variants discovered may promote the breeding process in the cattle industry.

OVARY FUNCTIONAL STATE IN FEMALE ADOLESCENTS WITH A HISTORY OF INTACT UTERINE APPENDAGES TORSION

The aim: To assess the functional state of gonads in female adolescents with a history of intact uterine appendages torsion (IUAT). Materials and methods: A total of 55 adolescent girls aged from 13 to 17 years were examined. 35 patients with a history of IUAT were divided into two groups: group 1 consisted of 21 patients with organ-preserving surgical treatment (detorsion) and group 2 of 14 patients with radical surgical treatment of IUAT. 20 healthy girls formed the control group. Assessment of the ovarian reserve was performed by the determination of serum anti-Müllerian hormone (AMH) level. Ultrasound scan was used to evaluate ovaries volume, number and diameter of antral follicles. Results: The AMH level in control group (2.4 ± 0.29 ng/ml) has no statistically significantly differences compared with group 1 (2.2 ± 0.35 ng/ml, p = 0.12) and differed significantly compared with group 2 (0.96 ± 0.11 ng/ml, p &lt; 0.05). No statistically significant differences in ovary volume were found between group 1 and control group (6.02 ± 0.21 cm3 and 6.14 ± 0.17 cm3, respectively, p = 0.125). The number and diameter of antral follicles in group 1 didn`t significantly differ from the control group (3.85 ± 0.35 and 4.00 ± 0.25, p = 0.116, 3.57 ± 0.42 mm and 3.70 ± 0.31 mm, p = 0.128, respectively). Conclusions: The method of choice for IUAT treatment taking into account upcoming motherhood is laparoscopy with detorsion. Dynamic ultrasound results and determination of AMH level indicate the organ-preserving tactics for surgical treatment of IUAT.

Effects of experimental exposure to zearalenone on reproductive system morphometry, plasma oestrogen levels, and oocyte quality of beef heifer.

The aim of the present study was to evaluate the effects of zearalenone (ZEA) on the reproductive system morphometry, oestrogen (E2) levels and oocyte quality of beef heifers. Twenty non-pregnant purebred Nellore (Bos indicus) heifers [age, ≥18months; initial body weight, 348±30kg (mean±standard deviation)] were used. The animals were randomly divided into experimental group and a control group of 10 animals each. Group experimental was administered 300ppb ZEA per os for 98days, and the control group was administered placebo per os for 98days. The administration of ZEA was carried out daily by adding mycotoxin to the diet. All heifers were evaluated weekly via rectal ultrasound examinations (12weeks). Diameters of the right and left uterine horns, right and left ovaries, largest antral follicle and corpus luteum were measured. Vulva size was also measured. Blood samples were collected to estimate E2 levels. At the end of 12weeks, the heifers were slaughtered, and the ovaries were sent to the laboratory for in vitro embryo production. A completely randomized design was adopted, and repeated measures analysis of variance (p<.05) was performed (except for oocyte quality). Vulva size (p=.0985); diameters of uterine horns (p=.0522), ovaries (p=.6955), antral follicles (p=.6355) and corpus luteum (p=.3808); and E2 levels (p=.3379) were not affected by the treatments. ZEA-contaminated diet significantly reduced (p=.05) the proportion of viable oocytes (49.4%, n=207) compared with the control diet (59.9%, n=222); however, the blastocyst rate did not differ between the groups (p=.9418). The results indicate that contamination of beef heifer's diet with 300ppb ZEA affected neither morphometric parameters nor plasma oestrogen levels; however, ZEA contamination was detrimental to oocyte quality.

Insulin-like growth factor 2 is produced by antral follicles and promotes preantral follicle development in macaques†.

Insulin-like growth factors (IGFs) are known for their involvement in endocrine and paracrine regulation of ovarian function. Although IGF2 is the predominant circulating and intraovarian form of IGFs in primate species, the stage-specific follicular expression, action, and regulation of IGF2 are not well defined. Therefore, experiments were conducted to investigate the follicular IGF production in response to steroid hormone regulation and the direct IGF actions on follicular development and function in vitro. Preantral follicles were isolated from rhesus macaque ovaries and cultured to the antral stage in media supplemented with follicle-stimulating hormone and insulin. Follicles were randomly assigned to treatment groups: (a) control, (b) trilostane (a steroid synthesis inhibitor), (c) trilostane + estradiol, (d) trilostane + progesterone, and (e) trilostane + dihydrotestosterone. Media was analyzed for IGF concentrations, which were correlated to follicle growth. Follicles produced IGF2, but not IGF1, at the antral stage. Steroid depletion decreased, whereas steroid replacement increased, IGF2 production by antral follicles. Media IGF2 levels correlated positively with antral follicle diameters. Macaque preantral follicles and granulosa cells were subsequently cultured without (control) and with recombinant human IGF2 supplementation. Follicle survival, growth, and paracrine factor production, as well as granulosa cell proliferation and gonadotropin receptor gene expression, were assessed. IGF2 addition increased follicle survival rates, diameters and inhibin B production, as well as granulosa cell proliferation. These data demonstrate that IGF2 produced by antral follicles, in response to steroid hormone regulation, could act as a paracrine factor that positively impacts preantral follicle development and function in primates.

Effect of Water Clover (Marsilea crenata) Ethanol Extracts on Follicle and Oocyte Diameter of Goat: In Vitro Study

Water clover (Marsilea crenata) is one of the herbal plants that has been using in alternative medicine. It possesses pharmacologically active compounds like flavonoid, which has cellular activities such as antioxidant and estrogenic activity. This study aimed to evaluate the impact of water clover ethanol extract (WCE) at different concentrations on the growth of follicles and oocytes based on follicles and oocytes diameter, respectively, after six days of culture. This experimental study used 24 isolated antral follicles (2.5-3.2 mm), which were randomly divided into four groups including control (without supplemented WCE) and experimental groups that supplemented with different concentrations of WCE (21.6, 43.2, and 86.4 µg ml-1) in culture medium for six days culture. The diameter of follicles was measured on days 0, 3, and 6. Additionally, oocytes diameter was also measured on day 6. The results indicate that the mean diameter of antral follicles and oocyte diameter of WCE 43.2 µg ml-1 was significantly increase compared to the other groups (P≤0.05). According to our results, WCE exerts its effect on the growth of the antral follicle and oocyte based on follicles and oocytes diameter respectively in a dose-dependent manner after six days of the antral follicle cultured.  Keywords: antral follicle, flavonoid, in vitro, oocyte, water clover.

The regulation of the follicular synchronization and sensitivity of rats with PCOS by AMH during prolonged pituitary downregulation

The modified prolonged gonadotropin-releasing hormone agonist (GnRH-a) protocol lessens the ovarian hyperstimulation syndrome (OHSS) effect and improves the clinical pregnancy rate of women with polycystic ovary syndrome (PCOS) compared with the standard long GnRH-a protocol. However, the molecular basis of this process needs to be elucidated. Sprague Dawley (SD) female rats with letrozole-induced PCOS were divided into GnRH-a and blank groups. Rats in the GnRH-a group were given triptorelin for 11 days, whereas those in blank group were given an equal volume of 0.9% NaCl. Meanwhile, the changes in estrus cycle, hormonal profile, ovary index, ovarian histopathology and body weight were measured. The expressions of anti-mullerian hormone (AMH), type II receptor of AMH (AMHRII), and FSH receptor (FSHR) were taken as the indicators of follicular sensitivity. Changes of follicular counting and differences in antral follicle diameter at each stage were evaluated. The number of follicles from primordial to antral stages increased during downregulation and the differences in antral follicle diameter were reduced in the GnRH-a group, whereas no significant difference was found in the blank group. The results of Western blotting and ELISA indicated that the level of AMH in ovarian total protein and serum had a similar dynamic change in the GnRH-a group. The results of immunohistochemistry showed that follicular AMH, AMHRII, and FSHR significantly decreased in the GnRH-a group. Prolonged GnRH-a protocol can improve synchronization and sensitivity of follicular development by balancing the expressions of AMH, AMHRII, and FSHR among follicles at all levels, thereby achieving better therapeutic effect.

Effect of Chitosan on Histology of Reproductive Organs of Female Wistar Rats (Rattus norvegicus) Exposed to Acetate Lead

Lead is one of the pollutants widely spread in the environment because it is not easily decomposed. Lead can affect system functions such as the ovary and endometrium. Lead can trigger oxidative stress by reducing antioxidant enzymes and increasing Reactive Oxygen Species (ROS). Lead can also reduce Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH) levels by disturbing the hypothalamus. Chitosan is an antioxidant compound that can reduce the toxic effects of lead. The purpose of this study was to study the effects of chitosan administration on the diameter of antral follicles, the number of endometrial arterioles, and the thickness of endometrial rats after lead acetate exposure. This study was an experimental laboratory using a posttest-only control group design approach applied on 25 female rats aged 8 weeks old, body weight 125-175 grams. Lead and chitosan were given orally with a sonde. There were 5 groups, namely, negative control group (without any treatment), positive control group (lead 175mg/kg/BW), treatment group 1 (lead 175mg/kg/BW + chitosan 16mg/kg/BW, treatment group 2 (lead 175mg/kg/BW + chitosan 32mg/kg/BW), and treatment group 3 (lead 175mg/kg/BW + chitosan 64mg/kg/BW) for 30 days. The rats were sacrificed at proestrus phase, which was proven from vaginal swab. Observations were carried out using the Hematoxylin Eosin (HE) staining method. The observations were analyzed using One Way ANOVA and followed by Least Significant Differences (LSD) test. The results showed significant results (p-value &lt;0.05). Chitosan can increase the diameter of the antral follicle, increase the number of endometrial arterioles, and increase the thickness of endometrial rats exposed by lead acetate.

Genes expression and phenotypic differences in corpus luteum and cumulus cells of commercial line and piau breed gilts

We aimed to characterize the expression of angiogenesis-related genes in corpus luteum (CL) and cumulus cells (CC) during estrous cycle in gilts of different genetic groups, as well as to study the relation between gene expression and phenotypic data. Forty five gilts were used as follows: L1, Commercial Line 1 (Large White x Landrace x Duroc) (n = 15); L2, Commercial Line 2 (Large White x Landrace x Pietrain) (n = 15); and Piau, Piau breed gilts (n = 15). Estrus observation started from 120 days of age. After the second observed estrus females were slaughtered (n = 3) on days 3, 5, 10, 14 and 18 of estrous cycle (first day of estrous cycle as Day 0). CL sampling was performed on days 3, 5, 10 and 14 and collection of CC and follicular fluid on days 14 and 18. Follicular fluid was used for analysis of estradiol levels and CC and CL samples for analysis of angiogenesis-related genes expression, ANGPT-1/-2 and TEK in CC and MMP-2, VEGFA, VEGFR-1/-2, ANGPT-1/-2 and TEK in CL. Piau gilts showed lower ovulation rate than both L1 and L2 gilts (P < 0.05), lower number of large antral follicles (>6 mm) at 18 days than L2 gilts (P < 0.05), and smaller diameter of the largest follicles at 14 days than L1 gilts (P < 0.05). Piau and L2 gilts showed higher estradiol levels in follicular fluid on day 18. Expression of ANGPT-1 and -2 genes in CC did not differ among genetic groups neither among days of the estrous cycle, but TEK gene expression was higher in L1 than L2 gilts on day 18. Expression of VEGFA, VEGFR-2 and MMP-2 genes in CL did not differ among genetic groups and days of cycle, but VEGFR-1 expression was higher in Piau than L2 gilts on days 10 and 14, and it was higher in L1 than L2 gilts on day 14. The ANGPT-1/-2 and TEK genes expression in CL were significantly higher in Piau than L1 gilts on day 10. The ANGPT-2/ANGPT-1 gene expression ratio in CL was higher in L1 than Piau and L2 gilts at 14 days, suggesting a shorter luteal phase for L1 gilts. Results indicated differences among genetic groups for the pattern of the angiogenesis-related genes expression in CL along estrous cycles, which may be reflected in phenotypic traits such as ovulation rate, estradiol levels in follicular fluid and number and diameter of antral follicles.