Large Follicles Research Articles

MORPHOLOGICAL MECHANISMS OF RADIATION-INDUCED ADRENAL ADAPTATIONS AND THE LEVEL OF RADIONUCLIDE CONTENT IN THE WHITE-CHESTED HEDGEHOG IN A ZONE OF HIGH RADIOACTIVE CONTAMINATION

For the first time, anatomical, histological and morphometric criteria for radiation-induced damage to the thyroid gland of the river otter have been determined. The lobes of the thyroid gland do not undergo morphological changes,and they have a permanent elongated ribbon-like shape. The height of the follicular epithelium and the Brown index are among the signs that reflect the functional state of the thyroid gland. In old animals, the Brown index has the highest value - 19.49 ± 0.39 conventional units. Structural and adaptive changes indicate that the thyroid glands of the river otter in the radiation zone of habitat belong to the glands of the small follicular type of structure. With age, a few large follicles are found, and in the early gerontological period, also zones of disorganization of parenchymal structures with complete disappearance of the follicular structure of the gland or desquamation of the thyroid epithelium. In sexually mature animals, the specific activity of 137Cs in the thyroid gland is 0.78±0.17 kBq/kg.

Antimüllerian hormone levels and IVF outcomes in polycystic ovary syndrome women: a scoping review.

Antimüllerian hormone (AMH) is a homodimeric glycoprotein secreted by granulosa cells from primary to large antral follicles, and it plays an important role in the regulation of early follicle growth. It is considered a reliable marker of ovarian reserve and a predictor of ovarian response to controlled stimulation. Polycystic ovary syndrome (PCOS) is an endocrine condition that affects women of reproductive age worldwide, and it is associated with high levels of AMH. PCOS patients may have worse maturation and fertilization rates compared to normo-ovulatory women. Some studies have demonstrated a positive correlation between AMH levels and qualitative aspects of assisted reproduction treatment; but it is not clear whether high levels of both serum and follicular fluid AMH in PCOS patients correlate with in vitro fertilization outcomes. We ran this scoping review of the literature to address this specific question. We comprehensively searched the databases PubMed and Cochrane Library until January 2023. We found that higher AMH levels are associated with higher oocyte yield, but PCOS patients tend to have fewer mature oocytes and impaired embryo quality and implantation rates. Pregnancy rates, however, are not affected by AMH levels or laboratorial outcomes. We also found that higher AMH levels are associated with worse PCOS features.

Expression of VEGF165 and VEGF165b during ovarian follicular development

Objective: To investigate the role of vascular endothelial growth factor (VEGF)165a, VEGF165b, and VEGF receptor (VEGFR) in the development of bovine follicles. Methods: We cultured follicular cells that were collected from small, medium, and large sized bovine follicles with estrogen and measured the expression of VEGF, VEGFR2 and VEGF165b by Western blot analysis and immunofluorescence. Results: The expression of VEGF165 increased in all follicle sizes and the expression of VEGF165b was increased in the small and large follicles after culturing in an estrogen containing medium. The expression of VEGFR2 was increased in the medium and large follicles after culturing with estrogen for 96 h. VEGF165 was activated at 100 ng/mL estrogen in the large follicles for 96 h. In addition, VEGFR2 was upregulated in the medium and large follicles after treated with 100 ng/mL estrogen for 96 h. Conclusions: This evidence suggests that the expression of VEGF165 and VEGFR is associated with estrogen stimulation during the development of bovine follicles and in an autocrine or paracrine manner. This reveals an advantage during oocyte maturation in vitro.

MORPHOMETRIC CHARACTERIZATION OF THE STRUCTURAL COMPONENTS OF THE THYROID GLAND UNDER CONDITIONS OF MODELED THERMAL SKIN INJURY

The key organ of the peripheral endocrine system, the thyroid gland, shows the ability to adapt and restructure at diff erent levels of structural and cellular organization under the infl uence of various factors. The aim of the study was to determine morphometric changes in the parameters of the structural components of the thyroid gland. The experiment was modeled on 42 male laboratory mature white rats. The burn was modeled with copper plates heated in water at a temperature of 97-100 °C. The size of the aff ected area was 20 % of the animal’s body area, which, according to the current classifi cation, corresponds to a seconddegree burn. Changes in various parameters were studied by morphometric analysis of the obtained histologic sections. Statistical processing of digital data was carried out using the software «Excel» and «STATISTICA» 10.0 using parametric and nonparametric methods of data evaluation. Morphometrically, it was found that the organ contains follicles of three types – large, medium and small, and their quantitative ratio is 1.4: 4,7: 3,9. It was found that the average value of the area of a small follicle is (2342.5498.43) μm2, medium – (3722.53130.44) μm2, large – (5733.77203.67) μm2. The average value of thyrocyte height is (11.430.41) μm. The follicular- colloid index of the thyroid gland of intact animals is the highest for small follicles and is 1.940.08, for medium follicles – 1.81 0.04 and for large follicles – 1.730.05. The average value of the colloid accumulation index for small follicles is the lowest and is 2.600.12, for medium follicles – 4.18 0.19, and for large follicles – 6.270.29. At the light- optical level, on day 7 of the experiment, an increase in the number (1.58 times (p<0.001) compared to the intact index) of large follicles was noted, which are located mainly on the periphery of the lobules and fi lled with a compacted colloid. A decrease in thyrocyte height (1.29 times (p<0.001) compared to the normal value), a signifi cant decrease in the follicular- colloid index and an increase in the colloid accumulation index detected in the early toxemia stage indicate a decrease in functional activity in the organ, disruption of adaptive processes in it and the onset of destructive changes. At the later stages of burn injury (14 and 21 days of the experiment), against the background of a statistically signifi cant (p<0.001) increase in endogenous intoxication, there is a development of deep destructive changes in thyrocytes, organ vessels, proliferation of connective tissue layers, impaired synthesis and secretion of hormones. These conditions are manifested by the dominance of large, overstretched follicles in the parenchyma of the thyroid gland, lined with fl at, degenerating thyrocytes, the height of which at day 21 is 3.31 times signifi cantly (p<0.001) lower than the intact value; a statistically signifi cant decrease in the follicular- colloid index; a signifi cant increase in the colloid accumulation index, as well as a signifi cant deviation from the normal level of pituitary- thyroid system homones, which indicates organ hypofunction.

Adipolin (C1QTNF12) is a new adipokine in female reproduction: expression and function in porcine granulosa cells.

Adipolin (C1QTNF12) has been described as a regulator of metabolism and is linked with the pathophysiology of PCOS. In this study, for the first time, we show the expression of C1QTNF12 in granulosa cells and its positive effect on porcine granulosa cell proliferation and steroid synthesis. Adipolin (C1QTNF12) is a recently discovered adipokine that plays an important role in glucose and insulin level regulation. Previous studies showed its reduced level in serum of women suffering from polycystic ovarian syndrome; however, whether C1QTNF12 regulates ovary function is still unknown. The aim of the study was first to determine the level of C1QTNF12 in the porcine ovarian follicles granulosa cells (Gc) and then its in vitro effect on proliferation and steroidogenesis as well as phosphorylation of several signalling pathways. Our results showed that the expression of C1QTNF12 was dependent on follicle size and was higher at the mRNA and protein level in Gc of small than large follicles from both prepubertal and mature animals. Similar pattern was observed for C1QTNF12 concentration in porcine follicular fluid. Additionally, we observed immunolocalisation of C1QTNF12 in Gc, theca cells and oocytes. We found that C1QTNF12 stimulated porcine Gc proliferation via the activation of protein kinase B (AKT). Moreover, C1QTNF12 enhanced progesterone, testosterone and oestradiol secretion by elevating STAR, CYP11A1, HSD3B and CYP19A1 mRNA expression and by activation of MAP3/1 pathway. Additionally, C1QTNF12 increased pMAP3/1-to-MAP3/1 protein expression ratio and enhanced IGF1-induced pTyr-IGF1Rβ-to-IGFR1β and pMAP3/1-to-MAP3/1 protein ratios. Taken together, C1QTNF12 could act directly on proliferation and steroid synthesis and serve as an important factor in in vivo ovarian follicle function, possibly regulating the course of folliculogenesis.

Effects of a mixture of glycerol monolaurate and cinnamaldehyde supplementation on laying performance, egg quality, and antioxidant status in laying hens.

This study aimed to determine the effects of a mixture of glycerol monolaurate and cinnamaldehyde (GCM) supplementation on the laying performance, egg quality, antioxidant capacity, and serum parameters of laying hens. A total of 1120 14-week-old Jingfen-1 strain laying hens with similar performance were randomly allocated to four dietary treatments: control, and GCM groups supplemented with 250, 500, or 1000 mg kg-1 for 12 weeks. Compared with the control group, GCM-supplemented groups significantly reduced (P < 0.05) the rate of unqualified eggs of laying hens aged 17-24 weeks. Supplementation of GCM significantly increased (P < 0.05) yolk color and serum glutathione peroxidase (GSH-Px) activity but decreased (P < 0.05) the hydrogen peroxide (H2 O2 ) content in the serum of laying hens at the age of 20 weeks. Furthermore, groups supplemented with GCM showed a significant increase (P < 0.05) in Haugh unit, yolk color, activities of total superoxide dismutase and GSH-Px, and the glucose content in serum, and a decrease (P < 0.05) in the content of urea nitrogen and H2 O2 and malondialdehyde in serum of laying hens at the age of 24 weeks. 500 mg kg-1 GCM supplementation significantly increased (P < 0.05) the number of large white follicles and 1000 mg kg-1 GCM supplementation decreased the number of large yellow follicles in 28-week-old laying hens. These results indicated that GCM supplementation has positive effects on reducing egg loss and improving egg quality in the early laying period of laying hens. © 2023 Society of Chemical Industry.

Влияние на размера на фоликулите и средите за култивиране върху ин витро матурацията на ювенилни свински ооцити

In vitro maturation (IVM) of oocytes is a critical step in the assisted reproductive technologies performed in pigs. The aim of the present study was to determine what effect the use of a particular maturation medium has on the maturation of oocytes from follicles of different sizes. The results of SF and LF oocytes cultured in medium containing FCS after 0, 28 and 46 h of IVM were compared. Before the start of culture (0 h), 92% of oocytes from both classes of follicles were in the GV stage and after the first half of IVM (28 h). At the end of culture (46 h), both groups of oocytes completed meiosis at MII, with only 83.0% of oocytes from SF compared to 94.0% from LF reaching this stage. The results of oocytes, cultured in medium containing PVA, after 0, 28 and 46 hours of IVM were compared. Before culture (0 h), 94% of oocytes from small follicles and 92.0% of those from large follicles were in the GV stage. After 28 hours, 90.0% (SF) and 92.0% (LF) of oocytes were in the Metaphase I stage of nuclear maturation. The majority of oocytes from both groups (SF 94.0%, LF 92.0%) underwent meiosis to Metaphase II after 46 h. The present study shows that oocytes from smaller follicles with deficiencies in meiotic competence have problems with cytoplasmic maturation that are potentiated by the use of a certain medium. It was found that COCs from SF under the influence of FCS showed stronger luteinization compared to COCs from LF, but this could be compensated by using serum-free maturation medium.

Dietary Canthaxanthin Supplementation Promotes the Laying Rate and Follicular Development of Huaixiang Hens.

Canthaxanthin(CX) is a ketocarotenoid, which is widely used in poultry production as a lipophilic antioxidant. Huaixiang chickens are a local breed in China famous for their excellent meat quality; improving their laying rate via nutritional regulation has attracted extensive attention. The aim of this study was to evaluate the effects of dietary CX on the laying rate and follicular development in Huaixiang hens. A total of 180 Huaixiang hens were randomly divided into five groups with six replicates, and six chickens per replication. The control group (CON) were fed a basal diet, and the treatment group (NT) were fed a basal diet supplemented with 4, 6, 8 and 10 mg/kg CX. All chickens were 26 weeks old, living at an average environmental temperature of 25 ± 2 °C with a relative humidity of 65-75%. The results showed that supplementing the CX improved the laying rate and large white follicles (LWF) number (p < 0.05) and increased the concentration of reproductive hormones (LH, FSH, E2 and Prog) (p < 0.05), and the basal diet supplemented with 6 mg/kg CX worked best. Moreover, CX could increase the activities of antioxidant enzymes SOD and GSH-Px (p < 0.05) and reduce the content of the lipid peroxidation product MDA in Huaixiang chickens (p < 0.05); again, 6 mg/kg CX was best. In conclusion, dietary CX had positive effects on the laying rate, ovarian structure, reproductive hormone secretion, follicle development, and the antioxidant capacity of Huaixiang hens, and 6 mg/kg CX was recommended to be added to the diet of Huaixiang chickens.

Dietary non-phytate phosphorus requirements for optimal productive and reproductive performance, and egg and tibial quality in egg-type duck breeders

Optimal dietary non-phytate phosphorus (NPP) is essential in poultry to maximise productive and reproductive performance, along with indices of egg and bone quality. This study aimed to establish the NPP requirements of egg-type duck breeders aged from 54 to 80 weeks on the following traits: egg production, egg incubation, egg quality, tibial characteristics, reproductive organ, plasma indices, and the expression of genes related to phosphorus absorption. Longyan duck breeders aged 54 weeks (n = 300) were randomly allotted to five treatments, each containing six replicates of 10 individually caged birds. Birds were fed corn-soybean meal-based diets containing 0.18, 0.25, 0.32, 0.38, and 0.45% NPP/kg for 27 weeks. The tested dietary NPP levels did not affect egg production or egg quality indices. The hatchling weight of ducklings increased (quadratic, P < 0.01) as dietary NPP level increased, and the highest value occurred with 0.25% NPP. The number of large yellow follicles (LYF), and the relative weights of LYF and ovary showed linear and quadratic responses to dietary NPP levels; the lowest number and relative weight of LYF occurred with 0.38% NPP, and the lowest ovarian weight was obtained with 0.25% NPP. There were no differences in tibial length, breaking strength, and mineral density in response to dietary NPP levels. In contrast, tibial content of Ca increased (linear, P < 0.01) with dietary NPP levels increasing from 0.18 to 0.45%, and the tibial content of P increased at 0.32% NPP and the higher dietary NPP levels. Plasma concentration of P showed a quadratic (P < 0.05) response to the dietary NPP levels, where the highest value was seen at 0.38% NPP. In conclusion, dietary NPP levels from 0.18 to 0.45% had no effects on egg production, and egg and tibial quality of duck breeders. The duck breeders fed a diet with 0.25% NPP showed the highest hatchling weight of their offspring, while those fed 0.38% NPP had the lowest number and relative weight of LYF. These results indicated that the diet with 0.25% NPP can be used in egg-type duck breeders to improve the hatchling weight of their offspring, without adverse effects on their productivity. The regression model indicated that the maximal hatchling weight of ducklings was obtained from duck breeders fed the diet with 0.30% NPP.

Ovarian follicle transcriptome dynamics reveals enrichment of immune system process during transition from small to large follicles in cyclic Indian Ghoongroo pigs

Ovarian follicular development is a critical determinant of reproductive performance in litter bearing species like pigs, wherein economic gains depend on litter size. The study aimed to gain insight into the differentially expressed genes (DEGs) and signalling pathways regulating follicular growth and maturation in Ghoongroo pigs. Transcriptome profiling of porcine small follicles (SF) and large follicles (LF) was conducted using NovaSeq600 sequencing platform and DEGs were identified using DESeq2 with threshold of Padj. < 0.05 and log2 fold change cut off 0.58 (LF vs. SF). Functional annotations and bioinformatics analysis of DEGs were performed to find out biological functions, signalling pathways and hub genes regulating follicular dynamics. Transcriptome analysis revealed 709 and 479 genes unique to SF and LF stages, respectively, and 11,993 co-expressed genes in both the groups. In total, 507 DEGs (284 upregulated and 223 downregulated) were identified, which encoded for diverse proteins including transcription factors (TFs). These DEGs were functionally linked to response to stimulus, lipid metabolic process, developmental process, extracellular matrix organisation along with the immune system process, indicating wide-ranging mechanisms associated with follicular transition. The enriched KEGG pathways in LF stage consisted of ovarian steroidogenesis, cholesterol and retinol metabolism, cell adhesion molecules, cytokine receptor interaction and immune signalling pathways, depicting intra-follicular control of varied ovarian function. The hub gene analysis revealed APOE, SCARB1, MMP9, CYP17A1, TYROBP as key regulators of follicular development. This study identified candidate genes and TFs providing steroidogenic advantage to LFs which makes them fit for selection into the ovulatory pool of follicles.

Dynamic regulation of semaphorin 7A and adhesion receptors in ovarian follicle remodeling and ovulation

The ovarian follicle is a complex structure that protects and helps in the maturation of the oocyte, and then releases it through the controlled molecular and structural remodeling process of ovulation. The progesterone receptor (PGR) has been shown to be essential in regulating ovulation-related gene expression changes. In this study, we found disrupted expression of the cellular adhesion receptor gene Sema7A in the granulosa cells of PGR−/− mice during ovulation. We subsequently found that expression of Sema7A in preovulatory follicles is promoted by gonadotropins and hypoxia, establishing an asymmetrical pattern with the SEMA7A protein enriched at the apex of large antral follicles. Sema7A expression was downregulated through a PGR-dependent mechanism in the periovulatory period, the abundance of SEMA7A protein was reduced, and the asymmetric pattern became more homogeneous after an ovulatory stimulus. Receptors for Sema7A can either repel or promote intercellular adhesion. During ovulation, striking inverse regulation of repulsive Plxnc1 and adhesive Itga5/Itgb1 receptors likely contributes to dramatic tissue remodeling. The adhesive receptor Itga5 was significantly increased in periovulatory granulosa cells and cumulus–oocyte complexes (COCs), and functional assays showed that periovulatory granulosa cells and COCs acquire increased adhesive phenotypes, while Sema7A repels granulosa cell contact. These findings suggest that the regulation of Sema7A and its associated receptors, along with the modulation of integrin α5, may be critical in establishing the multilaminar ovarian follicle structure and facilitating the remodeling and apical release of the cumulus–oocyte complex during ovulation.

Ovarian follicle size or growth rate can both be determinants of ovulatory follicle selection in mice†.

The endocrinology regulating ovulation of the desired number of oocytes in the ovarian cycle is well described, particularly in mono-ovulatory species. Less is known about the characteristics that make one follicle suitable for ovulation while most other follicles die by atresia. Bromodeoxyuridine (BrdU) injection was used to characterize granulosa cell proliferation rates in developing ovarian follicles in the estrous cycle of mice. This methodology allowed identification of follicle diameters of secondary (80-130μm), follicle-stimulating hormone (FSH)-sensitive (130-170μm), FSH-dependent (170-350μm), and preovulatory (>350μm) follicles. Few preovulatory-sized follicles were present in the ovaries of mice at estrus, the beginning of the cycle. Progressive increases were seen at metestrus and diestrus, when full accumulation of the preovulatory cohort (~10 follicles) occurred. BrdU pulse-chase studies determined granulosa cell proliferation rates in the 24-48h before the follicle reached the preovulatory stage. This showed that slow-growing follicles were not able to survive to the preovulatory stage. Mathematical modeling of follicle growth rates determined that the largest follicles at the beginning of the cycle had the greatest chance of becoming preovulatory. However, smaller follicles could enter the preovulatory follicle pool if low numbers of large antral follicles were present at the beginning of the cycle. In this instance, rapidly growing follicles had a clear selection advantage. The developing follicle pool displays heterogeneity in granulosa cell proliferation rates, even among follicles at the same stage of development. This parameter appears to influence whether a follicle can ovulate or become atretic.

Insulin regulates gap junction intercellular communication in porcine granulosa cells through modulation of connexin43 protein expression

Gap junction intercellular communication (GJIC) among granulosa cells plays an important role in folliculogenesis, and it is temporal-spatially regulated during follicular development. Connexin (Cx) proteins predominantly form the basal structure of gap junctions in granulosa cells. In our study, immunohistochemical analysis revealed that Cx43 is the most widely expressed connexin in porcine follicles, especially among the large antral follicles. With application of insulin on porcine granulosa cells, we found that insulin significantly facilitated the protein level of Cx43, not mRNA level. This process is dependent on the phosphorylated activities of AKT and Erk since selective AKT and Erk inhibitors, LY294002 and U0126, respectively, hampered the potential of insulin to up-regulate Cx43 protein expression. As a consequence, the insulin-enhanced Cx43-couple GJIC activity in porcine granulosa cells was corresponding attenuated by the administration of LY294002 and U0126. Our findings provide a new insight into the molecular mechanisms by which insulin mediates cell-cell communication in porcine granulosa cells and sheds light on nutrition-reproduction interactions.

Characterization of lesions of nutritional congenital goitre in cattle

There are few studies that classify and characterize the morphometric and immunohistochemical features of goitre in bovine thyroid glands (TGs). We investigated 39 bovine TGs (fetuses [9], stillbirths [18], neonates [12]) born to dams with low T4 hormone levels and no iodine supplementation and 10 (fetuses [3], stillbirths [3], neonates [4]) born to dams with normal T4 levels and supplemented with iodine. Body weight (BW), TG weight (TGW), TGW:BW ratio and histological lesions were determined. The TGs were classified histopathologically as normal gland (G0), mild goitre (G1), moderate goitre (G2) or severe goitre (G3). Various morphological and morphometric parameters were calculated from microscopic images using image analysis software. Immunohistochemistry was performed to detect proliferating cell nuclear antigen (PCNA). There were significant differences in the TGW:BW ratio among groups (P <0.05): 0.3 ± 0.1 in G0, 0.5 ± 0.3 in G1, 0.8 ± 0.3 in G2 and 1.3 ± 0.7 in G3. In G0, large homogeneous follicles with eosinophilic colloid were seen. In the groups with lesions (G1, G2 and G3), heterogeneity in follicle shape and size, height and area of thyroid follicular cells, height of thyroid follicular epithelium and PCNA immunolabelling were directly related to histopathological grade, with significant differences among groups (P <0.001), gradually increasing from G1 to G3 compared with G0. The TGW:BW ratio and histological grade were positively correlated (P = 0.008).

Cellular atlases of ovarian microenvironment alterations by diet and genetically-induced obesity.

Obesity, which can arise from genetic or environmental factors, has been shown to cause serious damages to the reproductive system. The ovary, as one of the primary regulators of female fertility, is a complex organ comprised of heterogeneous cell types that work together to maintain a normal ovarian microenvironment (OME). Despite its importance, the effect of obesity on the entire ovary remains poorly documented. In this study, we performed ovary single-cell and nanoscale spatial RNA sequencing to investigate how the OME changed under different kinds of obesity, including high-fat diet (HFD) induced obesity and Leptin ablation induced obesity (OB). Our results demonstrate that OB, but not HFD, dramatically altered the proportion of ovarian granulosa cells, theca-interstitial cells, luteal cells, and endothelial cells. Furthermore, based on the spatial dynamics of follicular development, we defined four subpopulations of granulosa cell and found that obesity drastically disrupted the differentiation of mural granulosa cells from small to large antral follicles. Functionally, HFD enhanced follicle-stimulating hormone (FSH) sensitivity and hormone conversion, while OB caused decreased sensitivity, inadequate steroid hormone conversion, and impaired follicular development. These differences can be explained by the differential expression pattern of the transcription factor Foxo1. Overall, our study provides a powerful and high-resolution resource for profiling obesity-induced OME and offers insights into the diverse effects of obesity on female reproductive disorders.

The effect of unexpected follicular development in artificial cycles on the clinical outcomes in frozen thawed embryo transfer based on propensity score matching

Objective: To study the effect of unexpected follicular development in artificial cycles on the clinical outcomes in frozen thawed embryo transfer based on propensity score matching(PSM). Methods: The retrospective cohort study analyzed the clinical data of 7 064 cycles (5 716 patients) of artificial cycle frozen-thawed embryo transfer (AC-FET) in the Reproduction Center of the Third Affiliated Hospital of Zhengzhou University from January 1, 2016 to December 31, 2020. The clinical data were divided into three groups according to the degree of follicular development in AC-FET: no follicular growth group (group A, 6 349 cycles), small follicular growth group (group B, 248 cycles), and large follicular growth group (group C, 467 cycles). Differences in clinical outcomes between the small follicle growth group (Group B) and the large follicle growth group (Group C) were compared with the no follicle growth group (Group A) after PSM and logistic regression to adjust for confounding factors at baseline. A binary logistic regression model was used to analyze the factors related to the unanticipated follicular development in AC-FET. Results: Age [M(Q1,Q3)] was [31.0 (28.0, 36.0)] years in Group A, [34.5 (30.0, 40.0)] years in Group B, and [36.0 (31.0, 41.0)] years in Group C. After adjusting for confounders, the differences between Groups A and B in clinical pregnancy rate (P=0.169), live birth rate (P=0.318), early abortion rate (P=0.470), and miscarriage rate (P=0.783) were not statistically significant. The differences in clinical pregnancy rate (P=0.743), live birth rate (P=0.486) and miscarriage rate (P=0.080) between Groups A and C were not statistically significant, while early miscarriage rate (P=0.034) differences were statistically significant. The age, BMI, basal AFC, AMH and starting dose of estrogen were correlates of the emergence of non-expected small follicles in Groups B and A. The adjusted OR (AOR) values (95%CI) were 1.03 (1.01-1.06), 0.93 (0.90-0.98), 0.97 (0.95-0.99), 0.96 (0.95-0.97), and 0.59 (0.45-0.77), all P<0.05. Age, basal AFC, AMH and starting dose of estrogen were the associated factors of the appearance of non-expected large follicles in Groups C and A. The AOR values (95%CI) were 1.03 (1.01-1.05), 0.93 (0.91-0.95), 0.96 (0.95-0.97), and 0.52 (0.42-0.64), all P<0.05. Conclusions: In AC-FET, the clinical outcome of small follicular growth is similar to that of unfollicular growth; Compared with the growth without follicles, the growth and development of large follicles can reduce the early abortion rate; Patients with older age, less AFC, lower AMH, and lower initial dose of estrogen could be more likely to have unanticipated follicular development during endometrial preparation.

Extracellular Vesicles Contribute to the Difference in Lipid Composition between Ovarian Follicles of Different Size Revealed by Mass Spectrometry Imaging.

Follicular fluid (FF) ensures a safe environment for oocyte growth and maturation inside the ovarian follicle in mammals. In each cycle, the large dominant follicle (LF) contains the oocyte designated to be ovulated, whereas the small subordinate follicles (SFs) of the same wave will die through atresia. In cows, the oocytes from the SF, being 2 mm in size, are suitable for in vitro reproduction biotechnologies, and their competence in developing an embryo depends on the size of the follicles. FF contains proteins, metabolites, fatty acids, and a multitude of extracellular vesicles (ffEVs) of different origins, which may influence oocyte competence through bidirectional exchanges of specific molecular cargo between follicular cells and enclosed oocytes. FF composition evolves along with follicle growth, and the abundance of different lipids varies between the LF and SF. Here, significant differences in FF lipid content between the LFs and SFs within the same ovary were demonstrated by MALD-TOF mass spectrometry imaging on bovine ovarian sections. We then aimed to enlighten the lipid composition of FF, and MALDI-TOF lipid profiling was performed on cellular, vesicular, and liquid fractions of FF. Differential analyses on the abundance of detected lipid features revealed specific enrichment of phospholipids in different ffEV types, such as microvesicles (MVs) and exosomes (Exo), compared to depleted FF. MALDI-TOF lipid profiling on MVs and Exo from the LF and SF samples (n = 24) revealed that more than 40% of detected features were differentially abundant between the groups of MVs and Exo from the different follicles (p < 0.01, fold change > 2). Glycerophospholipid and sphingolipid features were more abundant in ffEVs from the SFs, whereas different lysophospholipids, including phosphatidylinositols, were more abundant in the LFs. As determined by functional analysis, the specific lipid composition of ffEVs suggested the involvement of vesicular lipids in cell signaling pathways and largely contributed to the differentiation of the dominant and subordinate follicles.

Morphological features of the rat’s thyroid gland after application of defect in the tibiae

Aim to study the morphological features of the rats thyroid gland after application of a defect in their tibiae.&#x0D; Material and methods. Sixty white mature male rats were divided into two groups. Group I intact animals, Group II rats subjected to a through defect in the proximal metaphysis of the tibiae. The experiment time-line was: 3, 10, 15, 24, and 45 days. The qualitative features of the thyroid gland structure were studied using light and electron microscopy; the quantitative features with histomorphometry.&#x0D; Results. The large follicles lined with simple squamous epithelium on the periphery of the gland as well as large and medium-sized follicles lined with low simple cuboidal, less often squamous epithelium in the center were observed in rats of Group II on days 3, 10, 15 of the experiment. The colloid filled completely or most of the follicle; there were desquamated epitheliocytes and cells with signs of apoptosis. The height of the follicular epithelium of the thyroid gland was smaller than the values of the Group I by 8.44%, 5.52%, 3.86% and 5.53%, 3.70%, 3.25% from 3rd to 15th days; the area of nuclei of follicular cells by 5.22%, 4.93% from 3rd to 10th days and by 5.31% on 3rd day; nucleo-cytoplasmic ratio by 3.11%, 2.29%, 2.28% from 3rd to 15th days and by 4.63%, 3.01% from 3rd to 10th days in the central and peripheral parts of the gland respectively. On the 3rd day, electron-microscopically, thyrocytes were low cuboidal or flat shape, the nucleus had tortuous contours and accumulations of heterochromatin under the karyolemma and in the karyoplasm. The cisternae of the rough endoplasmic reticulum were collapsed or had small slit-like spaces. Single lysosomes and secretory granules in the apical part of the cells and short microvilli were visualized.&#x0D; Conclusion. The application of a defect in the tibiae of rats is accompanied by morphological changes in the thyroid gland from the 3rd to 15th days and then organ adapts to the trauma by the 24th and 45th days.

Is the Hippo Pathway Effector Yes-Associated Protein a Potential Key Player of Dairy Cattle Cystic Ovarian Disease Pathogenesis?

Cystic ovarian disease (COD) in dairy cattle is characterized by preovulatory follicles that become cysts, fail to ovulate and persist in the ovary; consequently, interfering with normal ovarian cyclicity. The intraovarian key players that orchestrate the alterations occurring in the preovulatory follicle and that culminate with cyst formation and persistence, however, remain uncertain. Interestingly, the Hippo pathway effector yes-associated protein (YAP) has been described in humans and mice as a key player of anovulatory cystic disorders. To start elucidating if YAP deregulation in ovarian follicle cells can be also involved in the pathogenesis of COD, we have generated a series of novel results using spontaneously occurring cystic follicles in cattle. We found that mRNA and protein levels of YAP are significantly higher in granulosa (GCs) and theca cells (TCs) isolated from cystic follicles (follicular structures of at least 20 mm in diameter) in comparison to respective cell types isolated from non-cystic large follicles (≥12 mm). In addition, immunohistochemistry and Western blot analyses used to determine YAP phosphorylation pattern suggest that YAP transcriptional activity is augmented is cystic GCs. These results were confirmed by a significant increase in the mRNA levels encoding for the classic YAP-TEAD transcriptional target genes CTGF, BIRC5 and ANKRD1 in GCs from follicle cysts in comparison to non-cystic large follicles. Taken together, these results provide considerable insight of a completely novel signaling pathway that seems to play an important role in ovarian cystic disease pathogenesis in dairy cattle.

Effect of use and dosage of p-follicle-stimulating hormone for ovarian superstimulation before ovum pick-up and in vitro embryo production in pregnant Holstein heifers

The benefit of ovarian superstimulation using exogenous FSH before ovum pick-up (OPU) and in vitro embryo production (IVEP) has been the subject of conflicting results. The objective of the present study, therefore, was to evaluate the effect of use and dose of porcine FSH (p-FSH) before OPU/IVEP on ovarian response and embryo production in pregnant heifers. Pregnant Holstein heifers (n = 48) were assigned randomly to receive 0, 160, or 300 mg NIH-FSH-P1 in a crossover design. Ovum pick-up was performed at 49, 63, and 77 d of gestation with a 14 d "washout" between OPU sessions. Follicle ablation was performed on D 0 (p.m.) and p-FSH treatments, consisting of 4 decreasing dose injections administered 12 h apart, were initiated 36 h after follicle ablation (D2 a.m.). Heifers underwent OPU on D 5 (a.m.), 40 h after the last p-FSH treatment, and cumulus oocyte complexes (COC) were subjected to IVEP procedures. Differences between treatment groups were evaluated using generalized linear mixed models. There were quadratic effects of treatment on both number and percentage of small (<6 mm), medium (6-10 mm) and large (>10 mm) follicles. Number and percentage of medium follicles increased with increasing p-FSH dosages, although the magnitude of the change was greater between 0 and 160 mg, than between 160 and 300 mg of p-FSH. Total number of follicles, number of COC recovered and number of viable COC increased linearly with increasing p-FSH dose. Conversely, there was no evidence for an effect of p-FSH dose on COC recovery percentage nor the percentage of viable COC. Cleavage percentage, number of cleaved oocytes, blastocyst percentage, and number of blastocysts increased linearly with increasing p-FSH dose. In conclusion, use of p-FSH before OPU resulted in greater superstimulatory response and oocyte competence which in turn increased IVEP. Furthermore, these effects were dose dependent such that use of a greater dose of p-FSH up to 300 mg progressively increased embryo yield.