The reduction of the m6A methyltransferase METTL3 in granulosa cells is related to the follicular cysts in pigs.

Follicular cysts are a common reproductive disorder in domestic animals that cause considerable economic losses to the farming industry. Effective prevention and treatment methods are lacking because neither the pathogenesis nor formation mechanisms of follicular cysts are well-understood. In this study, we first investigated the granulosa cells (GCs) of cystic follicles isolated from pigs. We observed a significant reduction in the expression of methyltransferase-like 3 (METTL3). Subsequent experiments revealed that METTL3 downregulation in GCs caused a decrease in m6A modification of pri-miR-21. This reduction further inhibited DGCR8 recognition and binding to pri-miR-21, dampening the synthesis of mature miR-21-5p. Additionally, the decrease in miR-21-5p promotes IL-1ß expression in GCs. Elevated IL-1ß activates the NFκB pathway, in turn upregulating apoptotic genes TNFa and BAX/BCL2. The subsequent apoptosis of GCs and inhibition of autophagy causes downregulation of CYP19A1 expression. These processes lower oestrogen secretion and contribute to follicular cyst formation. In conclusion, our findings provide a foundation for understanding and further exploring the mechanisms of follicular-cyst development in farm animals. This work has important implications for treating ovarian disorders in livestock and could potentially be extended to humans.

Contribution of the VEGF system to the follicular persistence associated with bovine cystic ovaries.

Cystic ovaries (CO) characterize a disorder frequently found in dairy cattle. However, despite the contributions by several researchers, the mechanism that leads to ovulatory failure has not yet been completely elucidated. Thus, the aim of this study was to examine the mRNA expression of bovine vascular endothelial growth factor (VEGFA)-164, VEGFA-164b and VEGF receptors (VEGFR1 and VEGFR2) by real-time PCR and protein expression by immunohistochemistry, immunofluorescence and Western blot in follicular fluid from dairy cows with spontaneous CO and in an experimental model of follicular persistence induced by prolonged treatment with progesterone. Results showed that both VEGFA isoforms and receptors were coexpressed in granulosa and theca interna cells and in follicular fluid of ovaries from all the groups evaluated. VEGFA-164, VEGFA-164b and VEGFR2 protein expression was higher in theca cells of persistent follicles from group P0 (expected time of ovulation) than in those from dominant follicles (as reference structure) from the control group (p < 0.05). Also, VEGFA-164 expression was higher in theca cells of cysts than in those of dominant follicles of the control group (p < 0.05). In follicular fluid, VEGFA-164 expression was higher in persistent follicles from group P5 (5 days of follicular persistence) than in the control, P0 and P15 groups, and higher in cysts than in dominant follicles from the control group (p < 0.05). This study provides evidence of an altered expression of VEGFA-164, VEGFA-164b and VEGFR2 during the formation of persistent follicles and cysts in cows. Together, these results evidence that early development of CO in cows is concurrent with an altered expression of these growth factors and that these alterations may contribute to the follicular persistence, angiogenic dysregulation and ovulatory failure found in cows with follicular cysts.

Association of RBP-4 gene polymorphisms with follicular cysts in large white sows.

Follicular cysts, which is a common infertility disease, can cause financial losses in pig breeding programmes. The pathogenesis and mechanisms of the formation of follicular cysts are not understood clearly. In our previous study, the concentration of retinol-binding protein 4 (RBP-4) in the follicular fluid (FF) of the ovary with follicular cysts was found to be significantly higher than that of normal ovary, thereby suggesting that RBP-4 may be a candidate biomarker for porcine follicular cysts. To study the association of RBP-4 and follicular cysts further, we detected the polymorphisms of the RBP-4 gene and the presence of follicular cysts by PCR-Restriction fragment length polymorphism (RFLP) assay. In this study, we screened the mutations of RBP-4 gene in 79 sows with follicular cysts and 100 normal sows without cysts. Results showed that +249-63G>C polymorphisms were significantly associated with follicular cysts, and sows with CC genotype in RBP-4 gene had a high risk of developing follicular cysts. Hence, our findings further proved that RBP-4 may be a novel biomarker for follicular cysts, which may be valuable for the diagnosis of follicular cysts and molecular breeding of pigs.

A new insertion/deletion fragment polymorphism of inhibin-α gene associated with follicular cysts in Large White sows.

Ovarian follicular cysts are anovulatory follicular structures that lead to infertility. Hormones play key roles in the formation and persistence of cysts. Inhibins are heterodimeric gonadal glycoprotein hormones that belong to the transforming growth factor-ß superfamily. These hormones suppress the secretion of follicle-stimulating hormone. In this report, partial fragment of inhibin-α (INHA) subunit gene of Large White pig was detected from the genomic DNA by polymerase chain reaction. The sequence showed a 283 bp fragment insertion/deletion (I/D) polymorphism in INHA subunit gene. A total of 49 Large White sows with cystic follicles and 152 normal sows were screened for this polymorphism. The relationship of INHA I/D polymorphisms with follicular cysts was investigated. The distribution of I/D was significantly different between cystic and normal sows, thereby suggesting that the INHA subunit gene might be a potential biological marker for breeding programs in pig.

Association of inhibin-α gene polymorphisms with follicular cysts in large white sows.

Ovarian follicular cysts are anovulatory follicular structures that have been identified in sows and are known to cause infertility. The pathogenesis of follicular cysts remains poorly understood. Hormones play key roles in the formation and persistence of cysts. The hormone inhibin is a member of the TGF-ß superfamily and is named for its negative regulation of FSH, another hormone that controls follicular recruitment and growth. In the present study, 48 sows with follicular cysts and 60 normal sows with no cysts were screened for mutations in the inhibin-α gene to examine the association of inhibin-α gene polymorphisms with the presence of follicular cysts. The results show that the c.-42G>A and c.3222G>A polymorphisms are significantly associated with follicular cysts and that sows with c.-42GG and c.3222GG genotypes have lower risk of developing cysts. Our findings may provide novel biological biomarkers and promising gene therapy candidates for follicular cyst formation in sows, which would greatly benefit pig breeding programs.

Lats1 Deletion Causes Increased Germ Cell Apoptosis and Follicular Cysts in Mouse Ovaries.

The Hippo signaling pathway is essential for regulating proliferation and apoptosis in mammalian cells. The LATS1 kinase is a core member of the Hippo signaling pathway that phosphorylates and inactivates the transcriptional co-activators YAP1 and WWTR1. Deletion of Lats1 results in low neonate survival and ovarian stromal tumors in surviving adults, but the effects of Lats1 on early follicular development are not understood. Here, the expression of Hippo pathway components including Wwtr1, Stk4, Stk3, Lats2, and Yap1 transcripts were decreased by 50% in mouse ovaries between 2 and 8 days of age while expression was maintained from 8 days to 21 days and after priming with eCG. LATS1, LATS2, and MOB1B were localized to both germ and somatic cells of primordial to antral follicles. Interestingly, YAP1 was predominantly cytoplasmic, whereas WWTR1 was nuclear in oocytes and somatic cells. Deletion of Lats1 caused an increase in germ cell apoptosis from 1.7% in control ovaries to 3.6% in Lats1 mutant ovaries and a 58% and 32% decrease in primordial and activated follicle numbers in cultured mutant ovaries. Surprisingly, there was an increase in Bmp15 but not Gdf9, Figla, Nobox transcripts or the somatic-specific transcripts Amh and Wnt4 in cultured Lats1 mutant ovaries. Last, Lats1 mutant ovaries developed ovarian cysts at a higher frequency (43%) than heterozygous (24%) and control ovaries (8%). Results showed that the Hippo pathway is active in ovarian follicles and that LATS1 is required to maintain the pool of germ cells and primordial follicles.

Hereditary trichilemmal cysts: a proposal for the assessment of diagnostic clinical criteria.

Trichilemmal cysts (TCs) can occur as sporadic lesions or in hereditary-familial settings with autosomal dominant transmission. These entities have not been widely analyzed in their peculiar aspects yet. The aim of this study was to describe a cohort of patients with diagnosis of TCs through a clinical and biomolecular characterization, intended to highlight some effective diagnostic criteria for their identification. Among 149 cases of this study, 24 cases of TCs (16.1%) arose in patients with at least one first-degree relative with diagnosis of TCs. Peculiar findings concerning hereditary lesions included the multiple presentation with an early onset age. On the basis of clinical evaluation, we propose a panel of clinical and histologic criteria for the diagnosis of hereditary TCs, which includes: (i) the diagnosis of TCs in at least two first-degree relatives or in three first- or second-degree relatives in two consecutive generations; (ii) at least one of the patients with TCs diagnosed <45 years; and (iii) the diagnosis of multiple or giant (>5-cm lesions) or rare histopathologic features (proliferating and ossifying) TCs.

[Perforating and ruptured trichilemmal cyst with metaplastic ossification]. / Perforierende rupturierte Trichilemmalzyste mit metaplastischer Ossifikation.

Trichilemmal cysts can occur as multiple scalp lesions and may have a familial history. Metaplastic ossification, as seen in several traumatic, inflammatory or neoplastic conditions, however, seems to be an exceedingly rare event in trichilemmal cysts and might be associated with previous rupture.

Quantitative and qualitative analysis of argyrophilic nuclear organizer regions in follicular cyst, keratocystic odontogenic tumor and ameloblastoma.

BACKGROUND: Multicystic ameloblastoma is a benign odontogenic tumor that exhibits a more aggressive behavior than keratocystic odontogenic tumor (KCOT) and follicular cyst. AIM: The purpose of the study was to evaluate the proliferation index nuclear organizer regions (NORs) and their distribution among the four odontogenic lesions with known different clinical invasive behavior. STUDY AND DESIGN: In a descriptive-analytical cross-sectional study, 60 paraffin blocks of odontogenic lesions were prepared for silver nitrate staining. MATERIALS AND METHODS: For the quantitative analysis, 100 cells were counted at ×100 and the mean value was calculated. The morphometric analysis of NORs showed that they can be distributed into normal (round to oval-shaped) and abnormal (large, bean-shaped and cluster-shaped) groups. One-way analysis of variance (ANOVA) and multiple comparison with Tukey test were used for the statistical analysis of the results. RESULTS: The argyrophilic NOR (AgNOR) numbers in multicystic ameloblastoma, unicystic ameloblastoma, KCOT, and follicular cyst were 7.4 ± 2.7, 6.1 ± 2.56, 4.7 ± 1.84, and 2.82 ± 1.052, respectively. The difference between ameloblastoma (unicystic and multicystic types) and either_KCOT, or follicular cyst was statistically significant (P<0.001) and, (P=0.001), respectively. In follicular cysts, normal AgNOR dots were not detected outside the nuclei. NOR histological patterns of KCOT were large, bean shaped and rarely cluster shaped and it was cluster-shaped in multicystic and unicystic ameloblastoma. CONCLUSION: The current study suggests that determination of clinical behavior of ameloblastoma in comparison with KCOT and follicular cyst in silver nitrate staining is related to higher proliferation activity and different NORs' distribution pattern. However, further clinical follow-up studies must be performed to prove this.

The cytobiological differences between two odontogenic cyst-lining keratinocytes.

PURPOSE: Odontogenic cysts are classified into a developmental group, including follicular cysts (FC) and keratocysts, and an inflammatory group including radicular cysts (RC). In clinical cases, we frequently encounter RC and FC. The purpose of this study was to investigate the cytobiological differences between two odontogenic cyst-lining keratinocytes using a cytobiological approach from the aspect of metabolic function and the degree of maturation of the epithelium. MATERIALS AND METHODS: Samples of odontogenic cyst-lining keratinocytes and oral keratinocytes collected at surgery, and of cultured oral keratinocytes, were analyzed (1) by immunohistochemical staining of granulocyte macrophage colony stimulating factor (GM-CSF), human beta defensin-2 (HBD-2) and chemokine receptor 6 (CCR6) expressing cell (Langerhans cell, helper T cell and suppressor T cell) antibodies, (2) by reverse transcription-polymerase chain reaction (RT-PCR) to determine the expression of GM-CSF and HBD-2 mRNA and (3) by gas chromatography to evaluate the composition of fatty acids (16:0, 18:2, 20:4) in the cell membranes of the keratinocytes. RESULTS: 1. Immunohistochemical staining indicated that HBD-2 and GM-CSF expression were higher in RC than in FC. 2. The same results were obtained from the RT-PCR analysis. 3. The % composition of palmitic acid (16:0) was significantly higher in the RC-lining keratinocytes (38.62±5.86%) and in the FC-lining keratinocytes (30.37±1.38%) than in the normal gingiva (23.00±1.40%). The % composition of essential fatty acids (18:2+20:4) was significantly higher in the FC-lining keratinocytes (26.20±3.55%) than in the RC-lining keratinocytes (20.50±8.17%). CONCLUSION: The present study demonstrated definite cytobiological evidence of the differences between RC and FC.

Follicular cyst in a patient with myotonic dystrophy: a case of cyst with differentiation toward follicular infundibulum, isthmus, inner root sheath, and hair.

We report a case of follicular cyst, which developed in a patient with myotonic dystrophy (MyD). Histopathologically, the cyst showed infundibular and trichilemmal keratinization, inner root sheath differentiation, aggregation of basaloid cells, and pilomatricoma-like changes in the pericystic connective tissue. These findings have been reported in follicular cysts with Gardner's syndrome (GS). Interestingly, pilomatricoma is known as one of the skin diseases associated with MyD, though there have been no reported cases of cyst formation with differentiation toward portions of hair follicle in a MyD patient. In our case, we hypothesized that the cyst might be derived from embryonic follicular germinative cells or follicular stem cells under the genetic influence of the MyD gene, as observed in follicular cysts in patients with GS.

Growth differentiation factor-9 is required during early ovarian folliculogenesis.

Growth factors synthesized by ovarian somatic cells directly affect oocyte growth and function, but it is unclear whether oocyte-secreted factors play a reciprocal role in modulating somatic cell functions in vivo. During the functional analysis of members of the transforming growth factor-beta superfamily in mouse development, we have uncovered a new family member, growth differentiation factor-9 (GDF-9), which is required for ovarian folliculogenesis. GDF-9 messenger RNA is synthesized only in the oocyte from the primary one-layer follicle stage until after ovulation. Here we analyse ovaries from GDF-9-deficient female mice and demonstrate that primordial and primary one-layer follicles can be formed, but there is a block in follicular development beyond the primary one-layer follicle stage which leads to complete infertility. Oocyte growth and zona pellucida formation proceed normally, but other aspects of oocyte differentiation are compromised. Thus, GDF-9 is the first oocyte-derived growth factor required for somatic cell function in vivo.

Comedöes familiares: estudo de três casos / Familial comedones: report of three cases

Apresentaçäo de três casos de comedöes familiares examinados no Hospital Universitário Antônio Pedro, Niterói, RJ, em 1992. Após uma revisäo literária, alguns dos aspectos mais interessantes säo discutidos