Number Of Follicles Research Articles

Effect of age on follicular dynamics of goat ovaries

Reproductive aging is influenced by a gradual decline in both the quantity and quality of oocytes found within ovarian follicles. In mammals, fertility relies heavily on the health of oocyte reserves, the dynamics of follicular development, and the functioning of the ovaries. This study aimed to investigate the impact of age on the follicular dynamics of goat ovaries. A total of 40 ovaries of Black Bengal goats were obtained from a local slaughterhouse and sorted into four age groups: 6-12 months, 13-15 months, 18-21 months, and 22-24 months. Subsequently, the ovaries were treated, and follicles were assessed through visually and histologically. The findings revealed that nearly all parameters, including weight, length, and width, of both ovaries in the 18-21 months age group were insignificantly different (p>0.05) from the other age groups. Histologically, the number of normal primordial and primary follicles was also insignificantly higher (p>0.05) in goats aged 18-21 months compared to the other age groups. Conversely, the quantity of secondary follicles significantly increased (p<0.05) with age, but in terms of antral follicles, a quadratic trend was observed, where they increased up to a certain age and then declined. Additionally, the results showed that the number of degenerated primary and antral follicles was significantly higher (p<0.05) in the ovaries of 22–24-month-old goats, while the number of degenerated secondary follicles was insignificantly higher (p>0.05) compared to the remaining age groups. In conclusion, age had an effect on the follicular dynamics of goat ovaries, suggesting that ovaries from goats aged 18-21 months, sourced from slaughterhouses, hold potential for quality follicles and oocytes that can be valuable for further research. Bangladesh Journal of Animal Science 52 (4): 88-97

Effect of Coenzyme Q10a and Clomiphene Citrate for Ovulation Induction in Patients with PCOS as Compared to Clomiphene Citrate alone

To evaluate the effects of coenzyme Q10A and clomiphene citrate taken in addition to other drugs to stimulate ovulation in people with polycystic ovarian syndrome. The Department conducted the study of Obstetrics and Gynecology, Pakistan Ordinance Factory (POF), Wah Cantonment, Rawalpindi from Mar 2019 to Feb 2021 By using a lottery, all of the patients were split into two random groups. Group A was given Coenzyme Q10A 60mg daily from day 1 of the cycle thrice a day and clomiphene citrate 150mg/day from days 2-6 of the cycle, and Group B was given only clomiphene citrates 150mg/day from 2-6 day of cycle for 5 days. The follicular growth and maturation were observed on days 10, 12, and 14 of the cycle with transvaginal ultrasound. Ovulation was confirmed by measuring serum progesterone level on day 21 of the cycle. Patients were followed for 3 cycles.The mean number of mature follicles in Group A was 1.63+0.49 and 1.3+0.47 in Group B (p-value=0.01). The analysis of either of the groups' ovulation statuses revealed that in 70% of patients (n=21) in Group A and 36.67% (n=11) in Group B, ovulation was initiated (p value= 0.01). We concluded that the outcome of combined coenzymeQ10A and clomiphene citrate is significantly better for patients with polycystic ovarian syndrome; inducing ovulation is more effective with clomiphene citrate alone.

Comparison of effects of curcumin and beta-carotene on ovarian damage caused by cisplatin

Purpose: We aimed the potential protective effects of curcumin (CUR) and beta-carotene (BC) against cisplatin (CIS)-induced ovarian damage using histological and immunohistochemical methods.
 Materials and Methods: We used 56 female Wistar albino rats, divided randomly into seven groups. Control rats did not receive any treatment; the sham group was administered 1 ml/kg sesame oil by gavage; the CIS group 5 mg/kg CIS; the CUR group 200 mg/kg CUR; the BC group 100 mg/kg BC; the CUR + CIS group, CIS after administration of CUR; the BC + CIS group was pretreated with BC, then administered CIS. The ovaries of all groups were excised five days after the last application. We assessed histopathology and counted and classified follicles. Expression of anti-Mullerian hormone (AMH) and nuclear factor-kappa B (NF-κB) was detected immunohistochemically. Apoptosis was evaluated using the TUNEL method.
 Results: CUR and BC are protective against decreased numbers of primordial, primary, preantral, secondary, and tertiary follicles caused by CIS. Both Cur and BC reduced ovarian NF-κB levels in comparison to the control group, and AMH immunoreactivity was almost identical for the control and CUR + CIS groups. Apoptotic cell counts indicated that CUR exerts a stronger anti-apoptotic effect than BC.
 Conclusion: CUR has a protective effect against ovarian damage brought on by CIS and greater antioxidant and anti-inflammatory properties than BC.

Enhancement of Melanocortin-4 Receptor (MC4R) and Constancy of Kiss1 mRNAs Expression in the Hypothalamic Arcuate Nucleus in a Model of Polycystic Ovary Syndrome Rat

Background: Hypothalamic Melanocortin-4 Receptor (MC4R) and kiss1/kisspeptin systems play roles in reproductive processes. This study was conducted to evaluate changes in MC4R and kiss1 genes expression in the arcuate nucleus (ARC) of the hypothalamus and its relationship with polycystic ovary syndrome (PCOS) in rats. Materials and Methods: In the current experimental study, 24 female rats were randomly and equally allocated into nulliparous and primiparous groups and then were divided into two subgroups of PCOS and control. PCOS was induced by exposure to continuous light. Sex-related hormones were evaluated by radioimmunoassay or immunoradiometric assay. Expressions of MC4R and kiss1 gene in the ARC of the hypothalamus of the rats were evaluated by real-time PCR. Histomorphometric alterations of ovaries were compared between groups. Results: Number of tertiary follicles and their size and number of atretic follicles in the PCOS subgroups were more than those in the controls (P<0.05) whereas the number of secondary follicles and corpus luteum in the PCOS subgroups were lower than those in the controls (P<0.05). Antrum and total diameters of tertiary follicles in the PCOS subgroups were greater and granulosa layer diameter was lower than those in the controls (P<0.05). The MC4R mRNA expression in the PCOS subgroups was 6.5-fold in nulliparous and 3.5-fold in primiparous groups more than their controls’ pairs (P<0.05). However, parity did not affect the expression of MC4R gene (P>0.05).The kiss1 mRNA expression in the PCOS and control subgroups was not significantly different (P>0.05). Conclusion: Overexpression of MC4R gene after PCOS induction in the ARC of the hypothalamus may link to metabolic disorders of induced PCOS in the rats. However, alteration in the kiss1 mRNA expression after PCOS induction was not observed in the rats. [GMJ.2018;7:e1070]

The Effects of Hot Pepper Seeds Added to the Diet in Quails on the Morphology of the Oviduct and Ovary

The aim of this study is to reveal how red-hot pepper seeds added to the diet in quails change ovarian follicle dynamics and the morphological structure of the oviduct. For this reason, a total of 48 female Japanese quails (Coturnix coturnix Japonica) aged 14 weeks were used as a material. While red hot pepper (Capsicum annuum L.) seeds were added to the diet of the animals in the experimental group at the rate of 2%, 4%, and 6%, it was not added to the control group. At the end of the study, while the number of white follicles showed a statistically significant increase in all groups compared to the control group (P

Effect of dexpanthenol on cyclophosphamide-induced ovarian toxicity: a histological and molecular study in rats

Research questionDoes dexpanthenol work as an effective therapeutic agent against cyclophosphamide (CYC)-induced premature ovarian failure (POF) in rats? DesignA total of 28 female Wistar Albino rats were randomly divided into four groups (n = 7 per group). The POF and POF plus dexpanthenol groups were intraperitoneally administered CYC at an initial dose of 50 mg/kg, followed by 8 mg/kg for 14 days. The dexpanthenol and POF plus dexpanthenol groups were both intraperitoneally administered dexpanthenol at a dose of 500 mg/kg/day for 15 days. ResultsIn the group administered CYC, the following was observed: a decrease in the ovarian index; a decrease in the numbers of primordial, primary, secondary and antral follicles; an increase in the number of corpus luteum and atretic follicles; a decrease in proliferation cell nuclear antigen immunoreactivity; a significant reduction in anti-Müllerian hormone and oestradiol levels; and an increase in serum FSH levels compared with controls. Dexpanthenol, on the other hand, reversed these effects. Quantitative reverse transcription polymerase chain reaction analyses showed that dexpanthenol increased Bcl-2, Akt1, mTOR, Nrf2 and HO-1 in CYC-induced ovarian tissues, but decreased Bax, Cas3, Hsp27, Hsp70, and Hsp90. Dexpanthenol treatment has a potential for inhibiting the intrinsic apoptotic pathway and oxidative stress levels in ovarian tissues via the downregulation of the mRNA expression of heat shock proteins and the activation of Nrf2/HO-1 pathways. ConclusionsOur findings demonstrated that dexpanthenol is an effective agent against POF caused by CYC; however, further experimental and clinical data are needed to use it effectively.

Clinical case of laser ablation of lingual amygdala

Hypertrophy of the lingual tonsil is a condition in which the lingual tonsil, located at the base of the tongue, enlarges. This tonsil consists of clusters of follicles separated by a central groove. In children under 5 years old, the number of follicles usually ranges from 10 to 12, while in adults up to 40 years old, it can reach 35 to 40. This structure plays an important role in the immune system, helping the body fight infections and microbes. Like other tonsils in the throat, the lingual tonsil can sometimes cause problems, such as hypertrophy (enlargement) or chronic infections. The causes of lingual tonsil hypertrophy can be diverse. In some cases, it develops as a compensatory process after the removal of the palatine tonsils. The primary cause in adults may be chronic inflammation of the pharynx. Severe degrees of lingual tonsil hypertrophy are very rare but can lead to various symptoms, such as coughing, difficulty swallowing and breathing, a feeling of pressure in the area beneath the tongue, and other nonspecific complaints. These symptoms are not pathognomonic for lingual tonsil hypertrophy. The challenges in diagnosis and the lack of a standardized approach to surgical treatment require detailed consideration and description in each individual case. Sometimes, patients with lingual tonsil hypertrophy complain of coughing paroxysms. In such cases, mechanical irritation of the lingual tonsil with a probe can provide some assistance in diagnosis. In the context of this study, lingual tonsillectomy surgery was performed on a patient using a diode laser. The results obtained confirmed the high level of technological feasibility and appropriateness of using laser ablation for the treatment of lingual tonsil hypertrophy. Laser ablation is one of the possible surgical procedures for treating lingual tonsil hypertrophy. It can be an effective and technologically advanced method, although it requires experience and qualification of specialists.

Clinical case of laser ablation of lingual amygdala

Hypertrophy of the lingual tonsil is a condition in which the lingual tonsil, located at the base of the tongue, enlarges. This tonsil consists of clusters of follicles separated by a central groove. In children under 5 years old, the number of follicles usually ranges from 10 to 12, while in adults up to 40 years old, it can reach 35 to 40. This structure plays an important role in the immune system, helping the body fight infections and microbes. Like other tonsils in the throat, the lingual tonsil can sometimes cause problems, such as hypertrophy (enlargement) or chronic infections. The causes of lingual tonsil hypertrophy can be diverse. In some cases, it develops as a compensatory process after the removal of the palatine tonsils. The primary cause in adults may be chronic inflammation of the pharynx. Severe degrees of lingual tonsil hypertrophy are very rare but can lead to various symptoms, such as coughing, difficulty swallowing and breathing, a feeling of pressure in the area beneath the tongue, and other nonspecific complaints. These symptoms are not pathognomonic for lingual tonsil hypertrophy. The challenges in diagnosis and the lack of a standardized approach to surgical treatment require detailed consideration and description in each individual case. Sometimes, patients with lingual tonsil hypertrophy complain of coughing paroxysms. In such cases, mechanical irritation of the lingual tonsil with a probe can provide some assistance in diagnosis. In the context of this study, lingual tonsillectomy surgery was performed on a patient using a diode laser. The results obtained confirmed the high level of technological feasibility and appropriateness of using laser ablation for the treatment of lingual tonsil hypertrophy. Laser ablation is one of the possible surgical procedures for treating lingual tonsil hypertrophy. It can be an effective and technologically advanced method, although it requires experience and qualification of specialists.

Molybdenum nanoparticles as a potential topical medication for alopecia treatment through antioxidant pathways that differ from minoxidil

BackgroundHair loss is a common dermatological condition including various types such as alopecia areata, androgenetic alopecia, etc. Minoxidil is a topical medication used for treating hair loss, which is effective for various types of alopecia. However, minoxidil has limitations in treating hair loss, such as slow onset of action and low efficacy, and it cannot effectively inhibit one of the major pathogenic factors of hair loss - excessive oxidative stress. MethodsTransition metal elements with rapid electron transfer, such as molybdenum, have been extensively studied and applied for inhibiting oxidative stress. We established a mouse model for hair growth and intervened with nano-sized molybdenum, minoxidil, and a combination of both. The physicochemical properties of nano-sized molybdenum enabled it to mediate oxidative stress more quickly. ResultsThe results showed that nano-sized molybdenum can accelerate hair growth, increase the number of local hair follicles, and reduce the expression of oxidative stress-related molecules such as iNOS, COX2, and androgen receptors. The combination of nano-sized molybdenum and minoxidil showed an additive effect in promoting hair growth. ConclusionOur findings suggest that nano-sized molybdenum might be a potential topical medication for treating hair loss by inhibiting the oxidative stress pathway. Nano-sized molybdenum, alone or in combination with minoxidil, could be a promising therapeutic approach for patients with hair loss, particularly those who do not respond well to current treatments. Further clinical studies are warranted to confirm the efficacy and safety of this novel treatment.

Evaluation of a long-acting recombinant bovine FSH for multiple ovulation and embryo transfer in dromedary camels

The present study was conducted to test a new super-agonist recombinant bovine FSH (rbFSH) to induce superovulation (SOV) in dromedary camels. In experiment I, a single IM injection of 40, 60, 80, 100, or 120 µg rbFSH was administered (4 donors per group) to determine the effective dose resulting in acceptable multiple ovulation and embryo yield. Administration of 40 µg was ineffective, while 100 and 120 µg were associated with increased numbers of developed follicles, corpora lutea, and recovered embryos compared to administration of 60 and 80 µg. In experiment II, donors were divided into treatment groups to compare rbFSH with two conventional protocols for SOV. Donors received a single dose of 2000 IU eCG in combination with 400 mg porcine follicle-stimulating hormone (pFSH; Folltropin-V®; Group 1, n = 29) or 500 µg of pFSH with 100 µg of pLH (Stimufol®; Group 2, n = 16). Group 3 (n = 19) received a single dose of 100 µg rbFSH. No difference was found in the size and number of follicles per donor. Response time, ovulation rate, and the number of corpora lutea and recovered embryos per donor were similar in all groups. The number of medium-sized and transparent embryos decreased while the number of small-sized and semi-transparent embryos increased in Group 3 (rbFSH) compared to the other two groups. The pregnancy rate of the recipients at 10 days post-ET, at two months of gestation, and the rate of early pregnancy loss (EPL) did not differ among the groups. In conclusion, a single IM administration of 100 µg rbFSH induces a successful superovulation in dromedary camels and has the advantage of reducing stress associated with multiple FSH administration of the conventional protocols.

Paeoniflorin Alleviates Cisplatin-Induced Diminished Ovarian Reserve by Restoring the Function of Ovarian Granulosa Cells via Activating FSHR/cAMP/PKA/CREB Signaling Pathway.

Paeoniflorin (PAE) is the main active compound of Radix Paeoniae Rubra (a valuable traditional Chinese medicine and a dietary supplement) and exerts beneficial effects on female reproductive function. However, the actions of PAE on diminished ovarian reserve (DOR, a very common ovarian function disorder) are still unclear. Herein, our study investigated the effect and potential mechanism of PAE on DOR by using cisplatin-induced DOR mice and functional impairment of estradiol (E2) synthesis of ovarian granulosa-like KGN cells. Our data show that PAE improved the estrous cycle, ovarian index, and serum hormones levels, including E2, and the number of antral follicles and corpora lutea in DOR mice. Further mechanism results reveal that PAE promoted aromatase expression (the key rate-limiting enzyme for E2 synthesis) and upregulated the FSHR/cAMP/PKA/CREB signaling pathway in the ovaries. Subsequently, PAE improved the levels of E2 and aromatase and activated the FSHR/cAMP/PKA/CREB signaling pathway in KGN cells, while these improving actions were inhibited by the siRNA-FSHR and FSHR antagonist treatments. In sum, PAE restored the function of E2 synthesis in ovarian granulosa cells to improve DOR by activating the FSHR/cAMP/PKA/CREB signaling pathway, which exhibited a new clue for the development of effective therapeutic agents for the treatment of DOR.

Effect of follicular wave synchronization using a progesterone-releasing intravaginal device-based protocol on in vitro embryo production in Bos taurus cows subjected to 14-day intervals ovum pick-up

Follicular wave synchronization (FWS) before ovum pick-up (OPU) is one of the strategies used to improve the efficiency of in vitro embryo production (IVP). This study aimed to evaluate the effect of FWS on the total follicular number, cumulus-oocyte complex (COC) recovery, and in vitro embryo development in Angus cows (n = 33) subjected to OPU with 14-day intersession intervals. Additionally, it was also evaluated the presence of carryover effects given the short intersession interval used. The experiment was run as a 2-treatment (FWS vs. Control) x 2-period (1 vs. 2) crossover design. Animals in the FWS group received an intravaginal progesterone implant (1gr), estradiol benzoate (2 mg), and D-cloprostenol (150 μg) on day 0 and the OPU was performed on day 5. Control group animals did not receive any hormone treatment. The FWS increased the number of 6–10 mm follicles (P = 0.05), but it decreased the COC recovery rate (P < 0.01). The FWS did not affect the total or frozen embryo numbers (P = 0.49 and P = 0.17; respectively), but it increased the total blastocyst cell number (P < 0.01). A carryover effect was found on the total and < 6 mm follicles number (P = 0.10 and P < 0.01; respectively), and on the regular, atretic, viable, and total number of COC (P = 0.01, P = 0.08, P = 0.02 and P < 0.01; respectively). We concluded that the FWS increased the quality of embryos after OPU with 14-day intersession intervals in Angus cows and that this kind of OPU/IVP scheme enabled the existence of a carryover effect, especially on the follicle number and COC morphology.

Linagliptin in combination with insulin suppresses apoptotic unfolded protein response in ovaries exposed to type 1 diabetes.

Type 1 diabetes mellitus (T1DM) is one of the main causes of ovarian atresia, but its molecular effect on the ovaries is not fully understood. Accumulating evidence suggests that T1DM causes excessive endoplasmic reticulum(ER) stress and insufficient adaptive unfolded protein response that triggers proapoptotic signaling pathways in ovarian tissue. In addition, problems such as amenorrhea and infertility, which are frequently seen in women with T1DM, continue despite the intensification of insulin therapy and improvement of metabolic control. Therefore new, and adjunctive treatments for women with T1DM need to be explored. We aimed to examine how the use of linagliptin, which has blood sugar-lowering effects and high antioxidant activity, together with insulin affects the expression levels of proteins and genes that play a role in ERstress in type 1 diabetic mouse ovaries. Eighty-four Balb/C 6-week-old female mice were randomly divided into seven groups: control, vehicle, diabetes + insulin, diabetes + linagliptin, diabetes + linagliptin + insulin, diabetes + TUDCA, and diabetes + TUDCA + insulin. TUDCA (an inhibitor of ER stress) groups are positive control groups created to compare linagliptin groups in terms of ER stress. Linagliptin and TUDCA were given by oral gavage and 1U insulin was administered subcutaneously for 2 weeks. A significant decrease was observed in the MDA and NOX1 levels and the number of atretic follicles in the ovaries of the diabetes + linagliptin + insulin group compared to the diabetes + insulin group. The use of linagliptin and insulin increased the expression of pro-survival XBP1s transmembrane protein and decreased the expression of proapoptotic ATF4, pJNK1/2, cleaved caspase 12, and cleaved caspase 3 in mouse ovaries. Our study provides new therapeutic evidence that linagliptin administered in addition to insulin induces ER stress mechanism-dependent survival in ovaries with type 1 diabetes.

BCAS2 regulates oocyte meiotic prophase I by participating in mRNA alternative splicing.

Oocyte meiotic prophase I (MI) is an important event in female reproduction. Breast cancer amplified sequence 2 (BCAS2) is a component of the spliceosome. Previous reports have shown that BCAS2 is critical in male germ cell meiosis, oocyte development, and early embryo genome integrity. However, the role of BCAS2 in oocyte meiosis has not been reported. We used Stra8-GFPCre mice to knock out Bcas2 in oocytes during the pachytene phase. The results of fertility tests showed that Bcas2 conditional knockout (cKO) in oocytes results in infertility in female mice. Morphological analysis showed that the number of primordial follicles in the ovaries of 2-month-old (M) mice was significantly reduced and that follicle development was blocked. Further analysis showed that the number of primordial follicles decreased and that follicle development was slowed in 7-day postpartum (dpp) ovaries. Moreover, primordial follicles undergo apoptosis, and DNA damage cannot be repaired in primary follicle oocytes. Meiosis was abnormal; some oocytes could not reach the diplotene stage, and more oocytes could not develop to the dictyotene stage. Alternative splicing (AS) analysis revealed abnormal AS of deleted in azoospermia like (Dazl) and diaphanous related formin 2 (Diaph2) oogenesis-related genes in cKO mouse ovaries, and the process of AS was involved by CDC5L and PRP19.

The comparison between fixed versus degressive doses of medroxyprogesterone acetate combined with letrozole in patients of progestin-primed ovarian stimulation protocol: a propensity score-matched study.

To explore the cycle characteristics and pregnancy outcomes of progestin-primed ovarian stimulation (PPOS) using fixed versus degressive doses of medroxyprogesterone acetate (MPA) in conjunction with letrozole (LE) in infertile women by propensity score matching (PSM) analysis. A retrospective cohort study. Tertiary-care academic medical center. A total of 3173 infertile women undergoing their first in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment within the period from January 2017 to December 2020. A total of 1068 and 783 patients who underwent a fixed dose of MPA combined with LE and a degressive dose of MPA combined with LE protocols, respectively, were enrolled in this study. The freeze-all approach and later frozen-thawed embryo transfer (FET) were performed in both groups. Propensity score matching (1:1) was performed. The primary outcomes were the dosage of MPA and the incidence of premature luteinizing hormone (LH) surges. The secondary outcomes were the number of oocytes retrieved, the cumulative live birth rate (CLBR) and the fetal malformation rate. We created a perfect match of 478 patients in each group. The dosage of MPA, the LH serum level on the eighth day of stimulation, progesterone (P) level and LH level on the hCG trigger day were significantly higher in the LE + fixed MPA group than in the LE + degressive MPA group (52.1 ± 13.1 mg vs. 44.9 ± 12.5 mg; 5.0 ± 2.7 IU/L vs. 3.7 ± 1.7 IU/L; 0.9 ± 0.5 ng/ml vs. 0.8 ± 0.5 ng/ml; 3.3 ± 2.4 IU/L vs. 2.8 ± 1.9 IU/L; P < 0.01). The duration of Gn, the number of follicles with diameter more than 16mm on trigger day, the estradiol (E2) level on the hCG trigger day were lower in the LE + fixed MPA group than in the LE + degressive MPA group (9.7 ± 1.7 days vs. 10.3 ± 1.5 days; 5.6 ± 3.0 vs. 6.3 ± 3.0; 1752.5 ± 1120.8 pg/ml vs. 1997.2 ± 1108.5 pg/ml; P < 0.001). No significant difference was found in the incidence of premature LH surge, the number of oocytes retrieved, the number of top-quality embryos, clinical pregnancy rate (CPR), CLBR or fetal malformation rate between the two groups. The combination of a degressive MPA dose with LE proved effective in reducing the total MPA dosage with comparable premature LH surge and pregnancy outcomes in women undergoing the PPOS protocol.

Single-cell RNA-seq identified novel genes involved in primordial follicle formation.

The number of primordial follicles (PFs) in mammals determines the ovarian reserve, and impairment of primordial follicle formation (PFF) will cause premature ovarian insufficiency (POI). By analyzing public single-cell RNA sequencing performed during PFF on mice and human ovaries, we identified novel functional genes and novel ligand-receptor interaction during PFF. Based on immunofluorescence and in vitro ovarian culture, we confirmed mechanisms of genes and ligand-receptor interaction in PFF. We also applied whole exome sequencing (WES) in 93 cases with POI and whole genome sequencing (WGS) in 465 controls. Variants in POI patients were further investigated by in silico analysis and functional verification. We revealed ANXA7 (annexin A7) and GTF2F1 (general transcription factor IIF subunit 1) in germ cells to be novel potentially genes in promoting PFF. Ligand Mdk (midkine) in germ cells and its receptor Sdc1 (syndecan 1) in granulosa cells are novel interaction crucial for PFF. Based on immunofluorescence, we confirmed significant up-regulation of ANXA7 in PFs compared with germline cysts, and uniform expression of GTF2F1, MDK and SDC1 during PFF, in 25 weeks human fetal ovary. In vitro investigation indicated that Anxa7 and Gtf2f1 are vital for mice PFF by regulating Jak/Stat3 and Jnk signaling pathways, respectively. Ligand-receptor (Mdk-Sdc1) are crucial for PFF by regulating Pi3k-akt signaling pathway. Two heterozygous variants in GTF2F1, and one heterozygous variants in SDC1 were identified in cases, but no variant were identified in controls. The protein level of GTF2F1 or SDC1 in POI cases are significantly lower than that of controls, indicating the pathogenic effects of the two genes on ovarian function were dosage dependent. Our study identified novel genes and novel ligand-receptor interaction during PFF, and further expanding the genetic architecture of POI.

Morphofunctional changes in the reproductive organs of females against the background of the use of a phytopreparation (experimental study)

The paper presents the results of the assessment of morphological changes in the uterus and ovaries of laboratory animals against the background of the introduction of therapeutic doses of a multifunctional phytopreparation, which includes alcohol extracts from plants: white hellebore (Veratrum album), dioecious nettle (Urtica dioica), yarrow (Millefolii herba) and has rumination, uterotonic, hemostatic and anti-inflammatory effects. Therapeutic doses for cows were tested, which for rats amounted to 0.2 ml per head. The phytopreparation was adminitered subcutaneously three times, every other day. The control group rats were subcutaneously injected with a saline solution of sodium chloride (NaCI 0.9%) according to the same scheme. The methods of digital microscopy were used in the study of micropreparations. Morphometric analysis of the histoarchitectonics of the uterus revealed a statistically significant increase in the diameter of the cross-section of the uterus of female rats of the experimental group by 25.8% (p≤0.05) compared with the control group due to an increase in the epithelial by 24.5% (p≤0.05) and the endometrial lamina of the uterine mucosa by 32% (p≤0.05), which indicates on increasing the secretory activity of the epithelial cover of the uterus. In the experimental group of rats treated with phytopreparation, a significant increase of 46% (p&lt;0.05) in the number of primordial follicles was noted. There was also a tendency to increase the number of primary – by 36.3%, secondary – by 41.7%, tertiary – by 75% of follicles and yellow bodies – by 22.2% in rats of the experimental group compared with the control indicators. Quantitative indicators of the follicular composition of the ovaries of rats of the experimental group and an increase in the follicle aging coefficient by 60% (p≤0.05) indicate stimulation of the ovarian reserve against the background of the use of phytopreparation.

Vitrified turkey ovarian tissue cultured and assessed through gene expression patterns: A potential screening tool

Biobanking of turkey ovarian tissue has the potential to play a crucial part in preserving female genetics. To date, ovarian tissue has only been vitrified using a standard protocol, with immediate analyses after warming, therefore, long-term cryoinjury is unknown. Long-term cryoinjury was investigated here by in-ovo culturing, fresh (non-vitrified), a purposefully suboptimal poor vitrification (PV), and the standard vitrified (StV) protocol. Assessments were performed via cellular morphological changes and mRNA gene expression differences, immediately (day 0) or after 2, 4, or 6 days of in-ovo culturing. On day 0, the mRNA levels of heat-shock protein A2 (HSPA2) were lowest in the fresh tissue, and increased 5-fold in the StV treatment, and 18-fold in the PV treatment. Whereas, by day 6, growth determining factor 9 (GDF9) mRNA levels within the fresh tissue were over 3-fold and 21-fold higher than StV and PV treatments, respectively. After 6 days of in-ovo culture the follicle density was highest in the fresh ovarian tissue (4701 ± 950 #/mm3), followed by the StV (1601 ± 300 #/mm3), with PV having the lowest density (172 ± 145 #/mm3). This shows that although the density of follicles was higher in StV versus PV, a considerable number (∼65 %) were lost compared to the fresh treatment. Additionally, the HSPA2 expression could be an early screening tool, whereas GDF9 expression could be a late screening tool, used to assess turkey ovarian tissue vitrification protocols. We conclude that the StV protocol should be further optimized to try and improve follicle numbers post-warming.

Niosomal hesperidin attenuates the M1/M2-macrophage polarization-based hepatotoxicity followed chlorpyrifos -induced toxicities in mice

Chlorpyrifos(CPF) is a well-known hepatotoxic agent that has side effects on several organs. On the contrary, hepatic macrophages are crucial in maintaining liver tissue integrity. The main objective of this study was to evaluate the effects and possible mechanisms of niosomal hesperidin (Nio + Hesp), a flavanone glycoside found in citrus fruits, on M1-M2 liver macrophage polarization and inflammatory cells in the brain, liver, and ovarian tissues. Forty C57 mice were divided into CPF(3 mg/kg), Sham(Dimethyl sulfoxide 40 μL/kg), CPF + Hesp(100 mg/kg), and CPF + Nio + Hesp (100 mg/kg) groups. The activity of sera superoxide dismutase (SOD) and malondialdehyde (MDA), brain, liver, and ovary tissues changes, and M1-M2 liver macrophage polarization were evaluated by examining the expression of CD163 and CD68 genes. Hepatic lesions consisting of sporadic foci of coagulation necrosis, inflammatory cell reaction, and regenerative fibrosis were seen following CPF injection, reflected by significant overexpression of CD163 and CD68 genes. In comparison, Nio + Hesp declined the amount of cell apoptosis in the liver and downregulated CD163 and CD68 gene expression. Both Nio + Hesp and Hesp alleviated CPF-induced hepatotoxicity, however, Nio + Hesp was superior to hesperidin in the downregulation of the CD163 and CD68 gene expression. Even though a significant difference between hesperidin and Nio + Hesp was observed in the number of Graafian follicles, corpus luteum, and peri-antral follicles, no substantial difference was observed in primary follicles. The ameliorative effects of Hesp and Nio + Hesp may be at least in part due to their antioxidant and anti-inflammatory properties. These findings showed that both M1- and M2-macrophages contributed to the development of hepatic lesions induced by CPF and provided information about macrophage activation, indicating the importance of analysis of macrophage phenotypes for hepatotoxicity based on M1/M2-polarization which can be downregulated by niosomal nesperidin.

The Role of Cellular Senescence in Cyclophosphamide-Induced Primary Ovarian Insufficiency.

Young female cancer patients can develop chemotherapy-induced primary ovarian insufficiency (POI). Cyclophosphamide (Cy) is one of the most widely used chemotherapies and has the highest risk of damaging the ovaries. Recent studies elucidated the pivotal roles of cellular senescence, which is characterized by permanent cell growth arrest, in the pathologies of various diseases. Moreover, several promising senolytics, including dasatinib and quercetin (DQ), which remove senescent cells, are being developed. In the present study, we investigated whether cellular senescence is involved in Cy-induced POI and whether DQ treatment rescues Cy-induced ovarian damage. Expression of the cellular senescence markers p16, p21, p53, and γH2AX was upregulated in granulosa cells of POI mice and in human granulosa cells treated with Cy, which was abrogated by DQ treatment. The administration of Cy decreased the numbers of primordial and primary follicles, with a concomitant increase in the ratio of growing to dormant follicles, which was partially rescued by DQ. Moreover, DQ treatment significantly improved the response to ovulation induction and fertility in POI mice by extending reproductive life. Thus, cellular senescence plays critical roles in Cy-induced POI, and targeting senescent cells with senolytics, such as DQ, might be a promising strategy to protect against Cy-induced ovarian damage.