Ovarian Samples Research Articles

Molecular characterization of ovarian mesonephric-like adenocarcinoma: Insights from single-cell RNA sequencing and mitochondrial metabolism.

Ovarian mesonephric-like adenocarcinoma (MLA) is a rare malignancy with limited understanding of its molecular features and therapeutic vulnerabilities. Although similar to uterine MLA, its unique characteristics remain undefined. This study aimed to characterize ovarian MLA using single-cell RNA sequencing (scRNA-seq) and compare it with high-grade serous ovarian cancer (HGSOC). We analyzed the cellular and molecular heterogeneity of an ovarian MLA sample using scRNA-seq. Differential gene expression and pathway analyses were performed to identify unique molecular signatures and therapeutic targets. HGSOC scRNA-seq datasets were used for comparative analysis. Ovarian MLA demonstrated reduced heterogeneity, with a predominance of epithelial cells compared to HGSOC. Transcriptomic profiling revealed an upregulation of mitochondrial metabolism and lipid biosynthesis genes, indicating a metabolic shift toward oxidative phosphorylation. Gene enrichment and protein-protein interaction analyses identified distinct pathways, including mitochondrial biogenesis and dynamics, suggesting mitochondrial reprogramming. This study provides the first scRNA-seq-based molecular characterization of ovarian MLA, differentiating it from HGSOC. Findings suggest potential therapeutic avenues, with a proposed combination therapy targeting MAP kinase and PI3K/AKT/mTOR pathways. Validation in larger cohorts is necessary for clinical application.

UBE2J1 is identified as a novel plasma cell-related gene involved in the prognosis of high-grade serous ovarian cancer

BackgroundImmune cells within tumor tissues play important roles in remodeling the tumor microenvironment, thus affecting tumor progression and the therapeutic response. The current study was designed to identify key markers of plasma cells and explore their role in high-grade serous ovarian cancer (HGSOC).MethodsWe utilized single-cell sequencing data from the Gene Expression Omnibus (GEO) database to identify key immune cell types within HGSOC tissues and to extract related markers via the Seurat package. The effects of immune cell markers on prognosis were analyzed via univariate Cox regression, least absolute shrinkage and selection operator (LASSO) and gene set variation analysis (GSVA) of bulk sequencing data from The Cancer Genome Atlas (TCGA)-HGSOC cohort. Finally, the effects of key markers on HGSOC cells were evaluated via Cell Counting Kit-8 (CCK-8), Transwell, colony formation, wound healing, immunofluorescence and in vivo tumor growth assays.ResultsAt the single-cell level, we detected a significant increase in the proportion of plasma cells in HGSOC samples compared to that in normal ovarian samples. Within HGSOC tissues, these plasma cells were found to interact with CD8 + T cells, fibroblasts and endothelial cells. In addition, patients in the high-plasma cell-related score group had better survival rates and higher epithelial‒mesenchymal transition (EMT), apoptosis and immune scores. Moreover, univariate Cox and LASSO regression analyses revealed that ubiquitin-conjugating enzyme E2 J1 (UBE2J1) is a prognostic marker in HGSOC. Further functional studies revealed that overexpression of UBE2J1 promoted cell proliferation, invasion, migration and colony formation, whereas UBE2J1 knockdown attenuated the abovementioned cellular behaviors. Additionally, UBE2J1 overexpression promoted EMT, as evidenced by alterations in the protein expression levels of N-cadherin, snail family transcriptional repressor 2 (Slug), Twist family BHLH transcription factor 1 (Twist 1) and E-cadherin. Moreover, we found that UBE2J1 silencing was able to inhibit the tumor growth in vivo.ConclusionsOverall, this study elucidated the role of plasma cells and revealed UBE2J1 as a novel oncogene in HGSOC, uncovering new mechanisms related to HGSOC tumorigenesis and promising therapeutic targets for HGSOC patients.

Stromal PDGFR-beta expression is a prognostic factor in high-grade serous ovarian cancer patients but is it also predictive for response to antiangiogenic treatment?

ObjectiveIn advanced ovarian cancer, the majority of patients receive anti-angiogenic treatment with bevacizumab. However, its use is often associated with severe side effects, and not all patients benefit from the therapy. Currently, there are no reliable biomarkers to predict response to treatment. Given their role as key regulators of angiogenesis, platelet-derived growth factor receptor-beta (PDGFR-beta) and vascular endothelial growth factor receptor-2 (VEGFR-2) are promising candidates for predictive biomarkers. This study evaluates their potential.MethodsPDGFR-beta and VEGFR-2 expression was evaluated using immunohistochemistry in a tissue microarray assay including 391 ovarian tissue samples. Correlation analyses with clinical and histopathological parameters were performed in a homogeneous cohort of 199 high grade serous ovarian cancer samples (HGSOC).ResultsIn HGSOC, strong stromal PDGFR-beta expression was associated with significantly shorter overall survival compared to weak/moderate expression. The impact of stromal PDGFR-beta expression on patient survival was however not restricted to the subgroup of patients receiving therapy with bevacizumab, and therefore cannot be considered as a predictive factor. For VEGFR-2, no or weak protein expression was found in the majority of the tumor samples. Survival analyses showed a more favorable prognosis with no or weak VEGFR-2 expression.ConclusionsHigh stromal expression levels of PDGFR-beta correlate with shorter overall survival in HGSOC. Thus, stromal PDGFR-beta might serve as a prognostic biomarker. No predictive effect in response to bevacizumab therapy could be attributed.

Ameliorative Effect of Combined Placenta-Derived Mesenchymal Stem Cells plus Platelet-rich Plasma on Polycystic Ovarian Model in Rats: A Biochemical and Histological Study.

Polycystic ovary syndrome (PCOS) is a common cause of infertility in women, characterized by metabolic and hormonal irregularities. We investigated the effects of placenta-derived mesenchymal stem cells (PDMSCs) and platelet-rich plasma (PRP), as well as their combination on follicular development, hormonal profile, inflammatory parameters, and insulin resistance in a model of PCOS. In this study, 25 female Wistar rats were randomly allocated into five groups: Sham (given a dose of 1 mL of a 0.5% carboxymethylcellulose (CMC) solution), PCOS (administered 1mg/kg of letrozole (LTZ) dissolved in CMC for 21 days), PDMSC (treated with a single intraovarian dose of PDMSCs), PRP (treated with a single intraovarian dose of PRP), and a combined PDMSC and PRP-treated group. After two weeks, serum and ovarian samples were collected for biochemical and histological analyses. Our results demonstrated that the simultaneous administration of PDMSCs and PRP had a synergistic effect compared to monotherapy, leading to an increase in estradiol (E2) and follicle-stimulating hormone (FSH) serum levels, a decrease in luteinizing hormone (LH) and testosterone levels, as well as inflammatory factors. Moreover, the combined therapy was associated with significantly lower levels of the homeostatic model of insulin resistance (HOMA-IR), fasting insulin (FINS), and blood glucose (FBG) compared to monotherapy. The combined treatment also caused a significant reduction in cystic follicles and an elevated number of corpus luteum, primordial, primary, secondary, and antral follicles. In conclusion, the combination of PRP and PDMSCs may have an ameliorative effect on modifying metabolic abnormalities and accelerating ovarian regeneration in PCOS.

Spatial transcriptome reveals histology-correlated immune signature learnt by deep learning attention mechanism on H&E-stained images for ovarian cancer prognosis

BackgroundThe ability to predict the prognosis of patients with ovarian cancer can greatly improve disease management. However, the knowledge on the mechanism of the prediction is limited. We sought to deconvolute the attention feature learnt by a deep learning convolutional neural networks trained with whole-slide images (WSIs) of hematoxylin-and-eosin (H&E)–stained tumor samples using spatial transcriptomic data.MethodsIn this study, 773 WSIs of H&E-stained tumor sections from 335 patients with treatment naïve high-grade serous ovarian cancer who were included in The Cancer Genome Atlas (TCGA) Pan-Cancer study were used to train, and validate, and to test a ResNet101 CNN model modified with attention mechanism. WSIs from patients in an independent cohort were used to further evaluate the model.ResultsThe prognostic value of the predicted H&E-based survival scores from the trained model on patient survival was evaluated. The attention signals learnt by the model were then examined their correlation with immune signatures using spatial transcriptome. After validating the model with the testing datasets, pathway enrichment analysis showed that the H&E—based survival score significantly correlated with certain immune signatures and this was validated spatially using spatial transcriptome data generated from ovarian cancer FFPE samples by correlating the selected signature and attention signal.ConclusionsIn conclusion, attention mechanism might be useful to identify regions for their specific immune activities. This could guide future pathological study for the useful immunological features that are important in modulating the prognosis of ovarian cancer patients.

A comparative study to estimate three commercial products of human chorionic gonadotropin (hCG) on blood biochemistry, sexual hormones concentrations, reproductive indices, and larvae production of African catfish Clarias gariepinus brood-stock

Currently, almost all hatcheries that produce catfish seeds depend on hormone stimulations, application of human chorionic gonadotropin hormone (hCG) is considered the common method for commercial spawning of African catfish Clarias gariepinus, regardless of whether they use an artificial or semi-industrial hatching approach. Therefore, to maximise profit, the best type must be selected based on cost and effectiveness. The present study compared three common products of hCG on blood sexual hormone, reproductive performance, and fry production per kg female. Four groups were designed to be: Group1: Untreated group to (control), Group2, Group3, and Group4, the brood-stock was hormonally induced with Choriomon®, Chorionic Gonadotropin, and Epifasi®, respectively. Both males and females received the same single dose of hCG (1500 IU)/kg of biomass. 80 brood-stock (40 female + 40 male) were used and distributed in two replicates in 8 square concrete ponds (water volume: 2.4 m3) each tank was randomly stocked at a rate of (5 males + 5 females). A single dose of hCG was injected intramuscularly at an angle of 45 degrees above the lateral line towards the tail. 24 hr after the injection blood sampling was drawn and samples of testes and ovaries were collected to measure reproductive indices and analyse chemical composition. Results showed superiority in all indicators of reproductive physiology that related to reproductive between hCG-treated brood-stock and untreated group. Besides, brood-stock injected with Choriomon® was the best in fry production (34587 larvae/kg brood stock) followed by Chorionic Gonadotropin and Epifasi®, respectively, while the Chorionic Gonadotropin group had the highest dose cost for producing 1000 fry.

No Bacterial Biomass Detected in Tissue From Patients With Ovarian Cancer and Serous Tubal Intraepithelial Carcinomas Using 16S rDNA Sequencing.

The ovarian oncobiome is subject to increasing scientific focus, but a potential link between bacterial dysbiosis and ovarian carcinogenesis remains controversial. Our primary aim was to characterize the bacterial microbiota in epithelial ovarian cancer samples. Secondarily, we aimed to compare results from the bacterial microbiota in formalin-fixed, paraffin-embedded ovarian tissue samples from 194 patients with epithelial ovarian cancer, fallopian tube tissue samples from 16 patients with serous tubal intraepithelial carcinomas and in benign fallopian tube tissue samples from 25 patients. Bacterial abundance was investigated by means of 16S rDNA Next Generation Sequencing. The 16S rDNA sequencing run produced a very low number of bacterial reads. Only seven samples displayed bacterial reads above 5000. Validation of detected bacterial reads by qPCR was negative and indicative of results being background amplification. Our results indicate no amount of bacterial biomass in the ovarian cancer, benign fallopian tube and in the samples with serous tubal intraepithelial carcinomas. The findings underline the importance of validating results from bacterial microbiota studies with additional technical assays before any conclusion may be drawn.

Transcriptome Concordance Between Borderline Tumors and Endometrioid Carcinoma: An Integrative Genomic Analysis.

Borderline ovarian tumors (BOTs) differ from ovarian carcinomas in their clinical presentation and behavior, yet their molecular characteristics remain poorly understood. This study aims to address this gap by integrating whole-exome sequencing (WES) and RNA sequencing (RNA-seq) to compare BOTs with high-grade serous carcinoma (HGSC), endometrioid carcinoma (EC), and clear-cell carcinoma (CCC). To elucidate the molecular features of BOTs and evaluate their similarities and differences in comparison to HGSC, EC, and CCC. The study analyzed 44 ovarian tumor samples, employing WES to identify genomic alterations and RNA-seq to examine transcriptomic profiles. Comparative analyses were conducted to investigate the molecular relationships among the tumor types. The genomic analysis revealed that BOTs share significant similarities with EC. Furthermore, the transcriptomic data highlighted a novel and substantial similarity between BOTs and EC, suggesting deeper biological linkages, including potentially shared oncogenic pathways or tumor microenvironmental factors. These findings challenge traditional classifications and suggest a closer molecular alignment of BOTs with EC than previously understood. This study provides new insights into the molecular characteristics of BOTs, demonstrating their significant resemblance to EC at both the genomic and transcriptomic levels. These results underscore the potential need to reconsider the molecular classification of ovarian tumors and open new avenues for research into the pathogenesis and treatment strategies for BOTs.

Isolation of viable follicles from cryopreserved human ovarian tissue

Introduction: Ovarian tissue freezing is the most effective method to maintain fertility for immature girls and women diagnosed with cancer. Nonetheless, because of the chance that malignant cells might reappearing following tissue transplantation, it is crucial to isolate the follicles from the frozen-thawed ovarian tissue of these individuals and employ them in the process of in vitro maturation process or artificial ovarian framework. This study aimed to assess the application of neutral red (NR) vital dye alongside collagenase IA for effectively isolating viable follicles from the vitrified human ovarian tissue samples. Methods: Two categories existed: the category with NR and the group without NR. Chopped (0.5×0.5 mm) strips of vitrified-warmed ovarian tissue from 10 transsexual individuals were placed into two falcon tubes with HTCM and Collagenase IA (1mg/ml). Neutral red (NR) was introduced to one of the falcons. Follicles were then isolated mechanically. The morphology, size, and viability of the follicles were assessed. The condition of the follicles was evaluated using fluorescent staining methods involving Calcein-AM and Ethidium homodimer-I. The t-test method was used to evaluate the data. Results: The number of isolated follicles with Neutral Red (46.50±25/02) exceeded those without NR (6.6±5.58; P < 0/0001). Additionally, according to the morphological studies, a majority of the isolated follicles from the transsexual ovarian cortex were primordial (77.4%), and primary (21.12%) follicles, with only a small number of secondary follicles (1.4%) identified in these tissues. Live/dead staining verified the viability of isolated follicles by displaying a green hue. Conclusion: The finding indicates that combining collagenase I with the vital dye Neutral red significantly facilitates the of follicles from dense human ovarian tissue.

Positive Effect of Elevated Thawing Rate for Cryopreservation of Human Ovarian Tissue: Transcriptomic Analysis of Fresh and Cryopreserved Cells.

Ovarian tissue cryopreservation has been gradually applied. It is essential to elucidate the differences between cryopreserved and fresh ovarian tissue and to refine cryopreservation protocols for improved outcomes. To explore the transcriptomic differences between fresh ovarian tissue and tissue cryopreserved with an elevated thawing rate. Ovarian tissue samples were collected and cryopreserved (frozen and thawed) following RNA sequencing and histological evaluation. Three groups were formed: fresh tissue (Group 1), frozen tissue after quick thawing at 100 °C (Group 2), and frozen tissue after slow thawing at 37 °C (Group 3). KEGG analysis showed that in comparison with Group 1, DEGs in Group 2 were mainly enriched in the cortisol synthesis and ovarian steroidogenesis pathways, and DEGs in the cells of Group 3 were mainly enriched in the ovarian steroidogenesis pathway. GO analysis showed that compared to cells of Group 2, DEGs in Group 3 were primarily enriched in the SRP-dependent co-translational protein targeting pathway and co-translational protein targeting to the membrane. The results were formulated with a minimal difference in the histological evaluation of cells after quick and slow thawed tissue. Cryopreservation of ovarian tissue by the described method does not decrease follicle production but downregulates the ovarian steroidogenesis pathway, reducing estrogen and progesterone secretion. The quick thawing of ovarian tissue increases the proliferation and apoptosis pathways of cells.

Comparative effectiveness of vitrification and slow freezing after heterotopic transplantation of human ovarian tissues

BackgroundThe aim of this study was to compare the effectiveness of two different vitrification methods and slow freezing in terms of the recovery of endocrine function, follicular morphology and proliferation, apoptosis of stromal cells, and angiogenesis after heterotopic transplantation of human ovarian tissue.MethodsOvarian tissue from young women aged 29 to 40 was subjected to two vitrification methods and one slow freezing method. The thawed ovarian tissue was then transplanted into nude mice and divided into three groups (VF1 group, VF2 group, SF group) according to the different freezing methods. Ovarian tissue samples were collected at 4 and 6 weeks post-transplantation. The recovery of ovarian function was evaluated by observing the estrous cycle and measuring estradiol levels using Elisa. Histological evaluation was performed to assess the integrity of ovarian follicles. TUNEL assay was used to detect stromal cell apoptosis, and immunohistochemistry was conducted to evaluate follicular proliferation and tissue angiogenesis.ResultsAfter heterotopic transplantation, mice in the experimental groups exhibited restoration of the estrous cycle. Hormone levels showed an increasing trend in the vitrification groups. At 6 weeks post-transplantation, the VF2 group had significantly higher hormone levels compared to the VF1 group and the slow freezing (SF) group (P < 0.05). At 4 weeks post-transplantation, the proportion of normal follicles was higher in the VF2 group compared to the other two groups (P > 0.05), and at 6 weeks post-transplantation, the VF2 group was significantly higher than the SF group (P < 0.05) and slightly higher than the VF1 group. Immunohistochemistry analysis indicated a higher proportion of proliferating follicles in the vitrification groups compared to the slow freezing group (P > 0.05). CD31 expression was established in all groups at 4 and 6 weeks post-transplantation, with better results in the slow freezing group compared to the vitrification group. TUNEL analysis showed that stromal cell apoptosis was higher in the SF group compared to the vitrification group at 4 weeks post-transplantation (P < 0.05), while there was no significant statistical difference among the groups at 6 weeks post-transplantation.ConclusionsVitrification showed better results than slow freezing, with the VF2 group performing slightly better than the VF1 group. Considering the lower economic and time costs associated with vitrification, it may be more suitable for ovarian tissue cryopreservation in major research centers in the future.

Mitochondria-targeted antioxidant mitoquinone mitigates vitrification-induced damage in mouse ovarian tissue by maintaining mitochondrial homeostasis via the p38 MAPK pathway

ObjectiveOvarian tissue cryopreservation has become a promising alternative for fertility preservation in cancer patients, allowing ovarian tissue to be stored for future autotransplantation. Oxidative stress damage occurring during the cryopreservation process may impact tissue quality and function. This study aims to investigate the protective effects and potential mechanisms of Mitoquinone (MitoQ), a mitochondria-targeted derivative of the antioxidant ubiquinone, during the vitrification of ovarian tissue in mice.MethodsKGN cells were treated with various concentrations (0.1, 1, 10, and 50 μM) of MitoQ to determine the optimal concentration. Female ICR mice were divided into three groups: control, conventional vitrification, and MitoQ-supplemented vitrification. Ovarian samples were cryopreserved, thawed, and assessed for tissue morphology using Hematoxylin and Eosin (H&E) staining, and mitochondrial changes using immunofluorescence, transmission electron microscopy, and Western blot analysis. RNA sequencing (RNA-seq) was employed to explore potential protective mechanisms. Autotransplantation experiments were conducted, and the long-term effects of MitoQ on ovarian function were evaluated by counting follicle numbers through H&E staining and measuring serum estradiol and AMH levels using ELISA.ResultsMitoQ at 1 μM was found to be the optimal concentration for maintaining follicular morphology after vitrification. It effectively reduced mitochondrial oxidative damage, preserved mitochondrial morphology, and regulated the expression of mitochondrial dynamics proteins (Drp1 and Mfn2). RNA-seq and Western blot analyses revealed that MitoQ inhibited the p38 MAPK pathway, thereby reducing apoptosis. Additionally, autotransplantation experiments showed that MitoQ treatment significantly increased follicle counts, estradiol (E2), and anti-Müllerian hormone (AMH) levels compared to conventional vitrification.ConclusionsMitoQ effectively mitigates vitrification-induced oxidative damage, maintains mitochondrial homeostasis, and preserves both follicular reserve and endocrine function. These findings suggest that MitoQ is a valuable adjunct in ovarian tissue cryopreservation and could significantly improve fertility preservation outcomes for cancer patients.

Fractalkine/CX3CL1 and macrophage inflammatory protein- 1β/CCL4 activity in the rat ovary with induced ovarian hyperstimulation.

Fractalkine (CX3CL1) and macrophage inflammatory protein-1β (MIP-1β)/CCL4 play a role in chemotactic activity, immune response, and inflammatory response. We aimed to investigate the effects of fractalkine and MIP-1β in the development of ovarian hyperstimulation syndrome (OHSS) by considering the inflammatory response during ovulation. Two equal groups of 20 immature female rats were created. Given that one of the rats in the group died, the control group was made up of 9 rats. Group 1 (G1) (n=9): Control group; G2 (n=10): OHSS group. Rats in the G2 group were administered 10 IU FSH for 4 days and 30 IU human chorionic gonadotropin on the fifth day. At 34 days old, all rats were sacrificed, and blood and ovarian tissue samples were collected to measure CX3CL1, CX3CL1R, MIP- 1β, tumor necrosis factor-alpha (TNF-α), interleukin (IL-8), hypoxia-inducible factor (HIF-1α), and interferon-gamma (IFN-γ) levels. Immunohistochemical scoring was performed for CX3CL1 and CX3CL1R in other ovarian tissue. Rat and ovary weights and serum CX3CL1, CX3CLR1, HIF-1α, MIP-1β, TNF-α, IFN-γ and IL-8 levels were significantly higher in G2 than in G1. Tissue IL-8, TNF-α, CX3CL1, CX3CLR1, MIP-1β levels and CX3CL1 and CX3CLR1 immunoreactivity scores were significantly higher in G2 than in G1. CX3CL1 and MIP-1β contribute to the pathophysiology of OHSS by playing a role in the development of inflammation.

Molecular characterization of ovarian squamous cell carcinoma originating from mature teratoma.

Squamous cell carcinoma (SCC) of the ovary, an uncommon form of gynecologic cancer, typically originates from the malignant transformation of a pre-existing mature ovarian teratoma (MOT). However, due to its rarity, the molecular pathways driving its development are not well understood. To address this knowledge gap, we performed molecular inversion probe (MIP) array analysis and targeted sequencing of 275 cancer susceptibility genes on 11 ovarian SCC samples derived from MOTs. Additionally, we conducted the same molecular tests on two samples of ovarian metastases of SCCs that originated from primary sites outside the ovary, specifically, one from endometrial cancer and one from cervical SCC. Utilizing MIP arrays, we identified failures in meiosis I and II, as well as instances of endoreduplication within haploid ova, in five, two, and four samples of ovarian SCCs arising from MOTs, respectively. Notably, such alterations were absent in samples of ovarian metastases, implying that primary ovarian SCCs may derive from teratoma cells. Targeted sequencing identified TP53 as the most frequently mutated gene in ovarian SCCs, occurring in 82% of cases. This was followed by mutations in PIK3CA (36%), PTEN (27%), and KMT2D (27%). Furthermore, mutations in CDKN2A and copy number loss of 9p21.3 were observed in 54.5% of the cohort. In summary, our study elucidates the germ cell origin of ovarian SCC and provides a comprehensive analysis of its genomic landscape, which may assist in differential diagnosis and inform the development of targeted therapies with potential clinical benefits. KEY MESSAGES: Failures in meiosis I/II and endoreduplication found in primary ovarian SCCs. Ovarian SCCs may derive from germ cells in mature teratomas. Alterations absent in ovarian metastases from SCC aid differential diagnosis. TP53 mutations found in 82% of ovarian SCC cases. CDKN2A mutations and 9p21.3 loss observed in 54.5% of ovarian SCC cohort.

Differential expression of the SLC34A2 gene in different histological subtypes of ovarian carcinomas

BACKGROUND: Patients with ovarian carcinoma demonstrate different sensitivity to chemotherapy; therefore, to increase the effectiveness of treatment, it is necessary to take into account the characteristics of each patient's tumor, including the histological subtype. AIM: To search for new molecular markers of ovarian cancer by analyzing the expression of candidate genes, including BAX, SLC34A2, MUC16, CD300A, and XKR8, in ovarian carcinomas of different histological subtypes. MATERIAL AND METHODS: Analysis of the expression of the BAX, SLC34A2, MUC16, CD300A, and XKR8 genes in 33 carcinomas taking into account their histological subtypes was performed using real-time polymerase chain reaction. Tumor samples from patients with ovarian carcinoma were obtained from the Blokhin National Medical Research Center of Oncology (Moscow) and the Republican Clinical Oncology Dispensary (Kazan) and divided into groups by histological subtypes: serous of high (n=16) and low (n=6) grade malignancy, endometrioid (n=8) and mucinous (n=3). Additional analysis was performed using microarray data from the Gene Expression Omnibus open database to determine the expression of selected candidate genes in ovarian carcinomas of different histological subtypes. The dataset included 4 normal ovary samples and 95 ovarian carcinoma samples of different histological subtypes: serous (n=41), endometrioid (n=37), and mucinous (n=13). Statistical analysis of the data was performed using Prism software. Nonparametric Dunn's test was used to compare gene expression in several patient groups. RESULTS: The expression level of the SLC34A2 gene was increased in low-grade serous carcinomas (p=0.0257) compared to mucinous carcinomas. Using bioinformatics analysis, we found increased expression of the SLC34A2 gene in serous (p=0.0023) and endometrioid ovarian carcinomas (p=0.0355) compared to normal ovarian tissues. CONCLUSION: The SLC34A2 gene can be considered as a potential molecular marker for differential diagnosis of ovarian cancer histological subtypes and a target for therapy of patients with low-grade serous ovarian carcinoma.

Expression of GnRH, Kisspeptin, and Their Specific Receptors in the Ovary and Uterus in Deslorelin-Treated Late-Prepubertal Bitches.

In this study, the expression and localization of gonadotropin-releasing hormone (GnRH1) and kisspeptin (KISS1) and their specific receptors in canine ovarian and uterine tissues were investigated after the application of deslorelin acetate (Suprelorin®, 4.7 mg, Virbac, France) in the late prepubertal period. We hypothesized that prolonged treatment of prepubertal dogs with deslorelin would alter the expression of GnRH and kisspeptin genes in the uterus and ovaries. Ovarian and uterine samples of 25 dogs with an average age of 7.8 ± 0.2 months and from mixed breeds were used. Following implant insertion, dogs entered estrus (EST; n = 6); dogs without estrus (N-EST; n = 10) comprised the experimental groups. Nine dogs with placebo implants served as a control (CONT). Ovarian and uterine tissues were investigated for expression of GnRH1, GnRHR, KISS1, and KISS1R/GPR54 mRNA and protein by using IHC and RT-qPCR. In the uterus, expression of GnRH1 significantly decreased in response to deslorelin treatment in the N-EST, compared with the control group. Compared with CONT, KISS1R expression in ovarian samples was significantly lower in the EST group. Uterine protein expression of GnRH1 appeared weaker in N-EST than in CONT. While GnRH1-system members and KISS1 protein were localized in the follicles at various stages and stroma, no or only weak signals were detected for KISS1R in the ovarian samples. Deslorelin-mediated induction of puberty by changing the expression of some of the GnRH and KISS1-system members seems to have an effect on ovarian and uterine functionality. Deslorelin implants can, therefore, not be considered a valuable alternative to induce fertile estrus in late-prepubertal bitches. However, further studies with a larger number of animals are needed to clarify the effect of deslorelin-mediated induction of puberty.

Evaluation of BRIP-1 (FANCJ) and FANCI Protein Expression in Ovarian Cancer Tissue.

Background: Ovarian cancer is one of the most common cancers in women. Markers associated with ovarian cancer are still being sought. The aim of this study was to evaluate the expression of BRIP-1 (FANCJ) and FANCI proteins in ovarian cancer tissue and to assess these expressions in differentiating the described clinical features. Methods: The study enrolled 68 patients with ovarian cancer. The cohort was divided into a HGSOC (high-grade serous ovarian cancer) group and a non-HGSOC group, which represented ovarian cancer other than HGSOC. Immunohistochemical evaluation of FANCI and BRIP-1 (FANCJ) protein expression in ovarian cancer tissue samples was performed. All statistical analyses were performed using StatView software (Carry, NC, USA). Results: The FANCI protein mostly showed moderate positive and strong positive expression, while BRIP-1 protein mostly showed no expression or positive expression. Patients with lower expression of FANCI and BRIP-1 showed differences in the clinical stage of HGSOC, which was not observed in patients with higher expression of these proteins. In addition, patients with lower BRIP-1 expression showed differences in menopausal status, which was not observed in patients with higher expression of this protein. Conclusions: This study shows that FANCI protein is a marker associated with lower FIGO stage and histologically high-grade cancer in a group of all ovarian cancers and in non-HGSOC.

Effects of carvacrol on hormonal, inflammatory, antioxidant changes, and ovarian reserve in polycystic ovary syndrome in Wistar rats.

The study, which explored the effects of carvacrol on female reproductive health, particularly in terms of hormonal, inflammatory, antioxidant, and ovarian reserves in polycystic ovary syndrome, could significantly influence future treatments and clinical practice. The study, conducted on thirty-five female Wistar albino rats, was designed to mimic the conditions of polycystic ovary syndrome in humans. The rats were randomly assigned into five groups: control (C), vehicle (V), polycystic ovary syndrome (PCOS), carvacrol (CAR), and polycystic ovary syndrome+ carvacrol (PCOS + CAR). The following practices were applied to the groups during the study. Normal saline was administered to the C group, DMSO to the V group, letrozole (1mg/kg/day) to the PCOS group, carvacrol (20mg/kg) to the CAR group, and letrozole and carvacrol together to the PCOS + CAR group for twenty-one days. At the end of the administration, blood, and ovarian samples were taken for hormone analysis (E2, and AMH), inflammatory (TNF-α, IL-6), oxidant-antioxidant analysis (malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px), and catalase), and histopathological examinations. Ovary, and body weights were measured, and the ovary index was calculated. As a result, it was observed that carvacrol caused beneficial effects through inflammatory, and antioxidant mechanisms in PCOS.

Autologous Ovarian Tissue Transplantation: Preoperative Assessment and Preparation of the Patient.

Ovarian tissue cryopreservation (OTC) is an innovative and established fertility preservation method. More than 150 live births have been reported worldwide to date with the use of this strategy. OTC is one of the options to preserve fertility in prepubertal girls and for women who have time constraints and/or contraindications for ovarian stimulation for oocyte/embryo freezing before cancer treatment. The success rate of the ovarian tissue transplantation (OTT) depends on many interrelated factors. Therefore, preoperative evaluation and preparation of the candidate patients for the procedure are of paramount importance. In this review, our aim was to provide a guide for the clinicians, which demonstrates step-by-step assessment and preparation of the patients and ovarian tissue samples for transplantation. We searched for published articles in the PubMed database containing key words, such as OTT, OTC, preoperative assessment, primordial follicle density, and cancer, in the English-language literature until May 2024. We did not include abstracts or conference proceedings. OTT is still a developing method as an effective fertility preservation approach. It is essential to perform a thorough preoperative evaluation of the patient to improve the success rates of transplantation. Preoperative evaluation and preparation of women for ovarian transplantation surgery should include safety management to prevent reimplantation of malignant cells, transplanting ovarian tissue with minimum follicle loss and the decision of the best transfer site.

First detection of D181 genotype of infectious bronchitis in poultry flocks of Morocco

BackgroundThis paper reports the first pathological and molecular characterization of the novel variant of infectious bronchitis virus (IBV) D181 in poultry flocks in Morocco and Africa.MethodsThe study includes six poultry farms, involving three flocks of layers aged between 28 and 67 weeks and three broiler flocks aged 27, 39 and 42 days from different regions of Morocco. In all affected layer flocks, a severe drop in egg production with poor eggshell quality was reported. Necropsy of dead birds was carried out, and samples of trachea, lungs, oviduct, ovaries, and kidneys were fixed in 10% neutral buffered formalin for histopathologic examinations, while other portions were stored at -20 °C for molecular analysis. Real time RT-qPCR for IBV gene group was performed, and IBV variants were identified. Partial S1 gene sequences were amplified by conventional RT-PCR, sequenced, and aligned for phylogenetic and amino acid similarity analysis.ResultsNecropsy of dead birds revealed misshapen and hemorrhagic ovarian follicles with an edematous oviduct and severe reaction in the cecal tonsils. A caseous material accumulation in the sinus was noted in few birds. In contrast, the broiler flocks exhibited respiratory clinical signs such as difficulty in breathing, sneezing, tracheal rales, watery eyes and lethargy, associated with a decrease in feed consumption. Mortality in broiler ranged from 2 to 15%. Histopathological analysis of samples showed a lympho-plasmocytic inflammation in the oviduct, trachea, and lungs. Individual necrosis of epithelial cells, with sloughing of the bronchial epithelium and accumulation of desquamated cells with mucus in the airways, was observed in some birds. Partial S1 gene sequencing and phylogenetic analyses showed that the Moroccan strains were very closely related to D181 strains isolated in Dutch layers and breeders in 2018. Nucleotide sequence identities reached 90.9–95% with the Dutch isolates (strain CK/NL/D181/2018).ConclusionOur sequencing results demonstrate for the first time that the D181 IBV genotype is circulating in Moroccan poultry. These findings justify permanent monitoring of circulating strains in order to appropriately adjust vaccination strategies to align with the evolving field situation.