Inhibin Alpha Research Articles

6485 Unilateral Adrenal Metastasis From Unknown Primary Carcinoma Mimicking Primary Adrenocortical Carcinoma

Abstract Disclosure: S. Watanabe: None. S. Suzuki: None. K. Ishiwata: None. Y. Yamazaki: None. H. Sasano: None. K. Yokote: None. Background: Adrenal metastasis from carcinomas of the lung, breast, and melanoma are observed. However, isolated adrenal metastasis with unknown primary carcinoma origin has been rare, and the diagnosis is challenging. We encountered a case of unilateral adrenal tumor clinically diagnosed as non-functional primary adrenocortical carcinoma, but found to be a metastatic tumor on postoperative pathology. Clinical Case: A 68-year-old man was referred to our hospital for examination of an 8mm right adrenal incidentaloma. He underwent hormone therapy for prostate cancer four years ago, and the condition is in remission with no recurrence signs. Follow-up CT revealed that the adrenal tumor rapidly enlarged from 8 mm to 36 mm within six months, and its mean attenuation value was 38 HU. MRI showed that the adrenal tumor had a low signal intensity on T1 and a high signal on T2. At the same time, there is mediastinal lymphadenopathy and a 9-mm nodule in the lower lobe of the right lung, which was considered benign from the stable condition. FDG-PET/CT showed FDG uptake only in the right adrenal tumor (maximum standardized uptake value of 21.39). [1]23I-MIBG scintigraphy did not show any accumulations. The serum and urinary adrenal hormones were all within normal limits. Notably, tumor markers such as prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), α-fetoprotein (AFP), and cancer antigen 19-9 (CA 19-9) were all within normal ranges. Therefore, we clinically suspected non-functional primary adrenocortical carcinoma, and the laparoscopic right adrenalectomy was performed. The resected right adrenal lesion was encapsulated and measured 79×54×22 mm. On histological analysis, the nodule comprised tumor cells with eosinophilic cytoplasm, and abundant tissue infiltrating lymphocytes. Immunohistochemical studies revealed that the tumor was not of adrenal origin due to negative steroidogenic factor-1 (SF-1) and inhibin alpha staining. From these results, the adrenal tumor was diagnosed as adrenal metastasis from unknown primary carcinoma. The tumor cells were positive for cytokeratin19, CEA, and thyroid transcription factor-1 but negative for PSA, p63, cytokeratin 5/6, insulinoma-associated protein-1, and synaptophysin. Genetically, the adrenal tumor harbored genomic abnormalities in CDKN2A, TP53, NF1, PBRM1, and KEAP1 genes. Conclusion: Isolated adrenal metastasis without a detectable primary tumor is rare but should be considered a differential diagnosis in the case of an enlarged, non-functioning adrenal tumor. Adrenalectomy or tumor biopsy may be actively considered to diagnose and determine the optimal course of treatment. Presentation: 6/3/2024

Solid ovarian tumor: A diagnostic and management dilemma

Abstract Pure Sertoli cell tumors (SCTs) are an uncommon type of ovarian sex cord–stromal tumor. The variety of symptoms, solid in nature and different histopathological presentation make it difficult the proper diagnosis and optimal treatment regimens in such tumor. Due to nonspecific clinical appearance, diagnosis is mistaken by ultrasound as fibroid or uterine mass. Intraoperative surprises and suspicious for malignancy due to the solid nature of the tumor lead to surgical management dilemma. The diagnosis is confirmed by histopathological examination only. The prognosis is excellent as most are detected in the early stages and surgical management is often curative in most cases. In this case, a 22 year old unmarried female presented to us with irregular periods for 5 years. Her trans-abdominal ultrasound revealed a solid hypoechoic mass of 5 x 6 centimeter size, in close approximate to the uterus on the right side, with probable diagnosis of broad ligament fibroid. On operative laparoscopy, uterus and left ovary was normal, the mass was originated from the right ovary, which was removed and its histopathological and immunohistochemistry report shows elevated alpha inhibin and steroidogenic factor 1 (SF 1) (Sex cord stromal marker) which is suggestive pure Sertoli cell tumor. There was no family/personal history of malignancy. She was treated with surgical resection and remains clear of disease. Prognostic indicators for SCTs include degree and type of heterologous element differentiation. Thorough characterization of such elements is crucial for adequate diagnosis and treatment.

Association between INHA gene polymorphisms and litter size in Hainan black goats.

The inhibin alpha (INHA) gene is one of the important genes affecting the reproductive traits of animals. Hainan black goats are the main goat breed in Hainan Island (China), whose development is limited by low reproductive performance. However, the relationship between INHA gene and the reproductive performance of Hainan black goats is still unclear. Therefore, the purpose of this work was to explore the effect of INHA gene polymorphisms on the litter size of Hainan black goats. Single nucleotide polymorphisms (SNPs) of INHA were detected, and the genetic parameters and haplotype frequency of these SNPs were calculated and association analysis was performed for these SNPs with the litter size. Finally, the SNP with significant correlations to litter size was analyzed by Bioinformatics tools. The results showed that the litter size of individuals with the AC genotype at loci g.28317663A>C of INHA gene was significantly higher than those with the AA genotype. This SNP changed the amino acid sequence, which may affect the function of INHA protein by affecting its structure. Our results suggest that g.28317663A>C loci may serve as a potential molecular marker for improving the reproductive traits in Hainan black goats.

Impact of Morphology in the Genotype and Phenotype Correlation of Bilateral Macronodular Adrenocortical Disease (BMAD): A Series of Clinicopathologically Well-Characterized 35 Cases.

Bilateral macronodular adrenocortical disease (BMAD) is characterized by the development of adrenal macronodules resulting in a pituitary-ACTH independent Cushing's syndrome. Although there are important similarities observed between the rare microscopic descriptions of this disease, the small series published are not representative of the molecular and genetic heterogenicity recently described in BMAD. We analyzed the pathological features in a series of BMAD and determined if there is correlation between these criteria and the characteristics of the patients. Two pathologists reviewed the slides of 35 patients who underwent surgery for suspicion of BMAD in our center between 1998 and 2021. An unsupervised multiple factor analysis based on microscopic characteristics divided the cases into 4 subtypes according to the architecture of the macronodules (containing or not round fibrous septa) and the proportion of the different cell types: clear, eosinophilic compact, and oncocytic cells. The correlation study with genetic revealed subtype 1 and subtype 2 are associated with the presence of ARMC5 and KDM1A pathogenic variants, respectively. By immunohistochemistry, all cell types expressed CYP11B1 and HSD3B1. HSD3B2 staining was predominantly expressed by clear cells whereas CYP17A1 staining was predominant on compact eosinophilic cells. This partial expression of steroidogenic enzymes may explain the low efficiency of cortisol production in BMAD. In subtype 1, trabeculae of eosinophilic cylindrical cells expressed DAB2 but not CYP11B2. In subtype 2, KDM1A expression was weaker in nodule cells than in normal adrenal cells; alpha inhibin expression was strong in compact cells. This first microscopic description of a series of 35 BMAD reveals the existence of 4 histopathological subtypes, 2 of which are strongly correlated with the presence of known germline genetic alterations. This classification emphasizes that BMAD has heterogeneous pathological characteristics that correlate with some genetic alterations identified in patients.

Inhibin Alpha Expression in Human Tumors: A Tissue Microarray Study on 12,212 Tumors.

As a result of its expression in corresponding normal cell types, inhibin alpha (INHA) is used as an immunohistochemical marker for adrenocortical neoplasms and testicular or ovarian sex cord stromal tumors. However, other tumors can also express INHA. To comprehensively determine INHA expression in cancer, a tissue microarray containing 15,012 samples from 134 different tumor types and subtypes was analyzed by immunohistochemistry. INHA positivity was found in 72 of 134 tumor categories, including 26 categories with ≥1 strongly positive case. A moderate to strong INHA positivity was found in 100% of 37 granulosa cell tumors of the ovary, 100% of 43 other sex cord stromal tumors of the ovary/testis, 100% of 31 granular cell tumors, 78.5% of 28 adenomas, 44% of 25 carcinomas of the adrenal cortex, and 46.7% of 15 pancreatic acinar cell carcinomas. At least a weak INHA positivity was seen in <33% of cases of 46 additional tumor entities. In summary, these data support the use of INHA antibodies for detecting sex cord stromal tumors, granular cell tumors, and adrenocortical neoplasms. Since INHA can also be found in other tumor entities, INHA immunohistochemistry should only be considered as a part of any panel for the distinction of tumor entities.

Lrriq1 is an essential factor for fertility by suppressing apoptosis.

Leucine-rich repeats and IQ motif containing 1 (LRRIQ1) gene is reportedly associated with plasma inhibin B levels. However, the function of LRRIQ1 remains unknown. In this study, we generated Lrriq1 knockout mice (Lrriq1-/- mice) and examined the effects of LRRIQ1 on inhibin B and fertility. Lrriq1-/- mice were generated using CRISPR/Cas9 genome editing technology. The expression of Inhibin B was examined by Western blotting using a protein extracted from the testis of a 3-month-old male mouse. Mating experiments were conducted using 7-week-old Lrriq1-/- mice and wild-type (WT) mice to examine fertility. Sperm concentration and sperm motility were measured using 3-month-old male mice. Expression analysis of inhibin B revealed that Lrriq1-/- mice exhibited reduced mRNA and protein levels of inhibin alpha (Inha), which constitutes the α subunit. In the mating experiment, the litter size of Lrriq1-/- male mice was 4.3 ± 2.9, which was significantly lower than that of WT male mice (8.3 ± 1.3) (p < 0.001). No difference in sperm count was observed between Lrriq1-/- and WT male mice; however, sperm motility (%) was significantly reduced in Lrriq1-/- mice (48.4 ± 4.9) when compared with WT mice (70.2 ± 4.7) (p < 0.001). Based on TUNEL staining, the testes and epididymal sperm of Lrriq1-/- mice showed high numbers of apoptosis-positive cells. Lrriq1 knockout reduced sperm motility and litter size by inducing apoptosis of testicular germ cells and epididymal sperm.

The Impact of Activin A on Fetal Gonocytes: Chronic Versus Acute Exposure Outcomes.

Activin A, a TGFβ superfamily member, is important for normal testis development through its actions on Sertoli cell development. Our analyses of altered activin A mouse models indicated gonocyte abnormalities, implicating activin A as a key determinant of early germline formation. Whether it acts directly or indirectly on germ cells is not understood. In humans, the fetal testis may be exposed to abnormally elevated activin A levels during preeclampsia, maternal infections, or following ingestion of certain medications. We hypothesized that this may impact fetal testis development and ultimately affect adult fertility. Germ cells from two mouse models of altered activin bioactivity were analysed. RNA-Seq of gonocytes purified from E13.5 and E15.5 Inhba KO mice (activin A subunit knockout) identified 46 and 44 differentially expressed genes (DEGs) respectively, and 45 in the E13.5 Inha KO (inhibin alpha subunit knockout; increased activin A) gonocytes. To discern direct effects of altered activin bioactivity on germline transcripts, isolated E13.5 gonocytes were cultured for 24h with activin A or with the activin/Nodal/TGFβ inhibitor, SB431542. Gonocytes responded directly to altered signalling, with activin A promoting a more differentiated transcript profile (increased differentiation markers Dnmt3l, Nanos2 and Piwil4; decreased early germ cell markers Kit and Tdgf1), while SB431542 had a reciprocal effect (decreased Nanos2 and Piwil4; increased Kit). To delineate direct and indirect effects of activin A exposure on gonocytes, whole testes were cultured 48h with activin A or SB431542 and collected for histological and transcript analyses, or EdU added at the end of culture to measure germ and Sertoli cell proliferation using flow cytometry. Activin increased, and SB431542 decreased, Sertoli cell proliferation. SB431542-exposure resulted in germ cells escaping mitotic arrest. Analysis of FACS-isolated gonocytes following whole testis culture showed SB431542 increased the early germ cell marker Kit, however there was a general reduction in the impact of altered activin A bioavailability in the normal somatic cell environment. This multifaceted approach identifies a capacity for activin A to directly influence fetal germ cell development, highlighting the potential for altered activin A levels in utero to increase the risk of testicular pathologies that arise from impaired germline maturation.

Rectifying cow infertility under heat stress by immunization against inhibin and supplementation of progesterone

This study was undertaken for the development of novel techniques that are based on immunoneutralization of inhibin bioactivity to improve Holstein cow fertility. A series of 4 experiments were carried out on 2 farms that were located in subtropical or temperate regions, to test the effects of immunization against inhibin alpha subunit on cow fertility under varying degrees of heat stress conditions. Though immunization against inhibin alone improved conception rate (CR) after TAI moderately in cows under mild heat stress conditions, the treatment plus progesterone supplementation substantially enhanced CR in the range of 25 to 35 percentages from severe heat stress to comfortable weather conditions. There existed an additive effect between immunization against inhibin and progesterone supplementation that maximally enhanced CR. Further, immunization against inhibin increased both FSH and activin A concentrations in blood during both follicular and luteal phases. It also significantly increased blood concentrations of E2 in the follicular phase but decreased P4 concentrations during the early pregnancy. However, interferon-tau concentrations in blood around the time of pregnancy recognition were doubled in the inhibin immunized cows. In conclusion, immunization against inhibin plus P4 treatment enhances ovarian follicle and the subsequent early embryo developments that help to greatly improve the fertility of Holstein dairy cows.

Cytosine-phosphate-guanine oligodeoxynucleotides regulate the cell cycle, apoptosis, and steroidogenesis of mouse ovarian granulosa cells by targeting inhibin alpha (1 ~ 32) fragments.

Cytosine-phosphate-guanine oligodeoxynucleotides (CpG-ODNs), which exist in vertebrate, bacterial, and viral genomes, are regarded as strong immune adjuvants. To date, the biological activities of CpG-ODNs in reproduction remain unknown. Here, we investigated the effects of CpG-ODNs on the cell cycle, apoptosis, and steroidogenesis in mouse granulosa cells (mGCs), in combination with inhibin alpha (1 ~ 32) fragments. mGCs were transfected with pEGFP (containing green fluorescent protein, as a control), pEGISI (containing inhibin alpha (1 ~ 32) fragments), or pEGISI-CpG-ODNs (containing inhibin alpha (1 ~ 32) fragments and CpG-ODNs motifs) plasmid for 48h in vitro. Our results showed that the mRNA and protein expression levels of inhibin alpha were downregulated in mGCs transfected with pEGISI-CpG-ODNs, compared to those transfected with pEGISI. Flow cytometry demonstrated that pEGISI-CpG-ODNs transfection promoted cell proliferation (for example, increasing the number of cells in S and G2 phases) and decreased apoptosis, compared to pEGISI transfection. The present study also indicated that the expression of cell cycle-related genes (cyclin D2, cyclin D3, cyclin E1, Cdk2, and Cdk6) was increased, while the expression of apoptosis-related factors (Fas, FasL, caspase-8, and caspase-3) decreased after pEGISI-CpG-ODNs treatment. Additionally, pEGISI-CpG-ODNs reversed the effect of pEGISI on the secretion of estradiol in mGCs, which was further validated by upregulating the levels of its synthesis-related factors (StAR, Cyp11a1, and 17β-HSD II). Nevertheless, pEGISI-CpG-ODNs or pEGISI did not affect the concentration of progesterone nor changed the expression levels of its synthesis-related factors (3β-HSD I and Cyp19a1). In conclusion, this study demonstrated that CpG-ODNs may affect the cell cycle, apoptosis, and steroidogenesis by targeting the effects of inhibin alpha (1 ~ 32) fragments, supporting the potential role of CpG-ODNs in the development of granulosa cells.

Primary pleomorphic liposarcoma of the liver: a case report and literature review

BackgroundPrimary liposarcoma arising from the liver is exceedingly rare. There have been very few reports documenting primary hepatic liposarcoma, especially of the pleomorphic subtype. Surgery is currently the only established treatment method, and the prognosis remains poor. In this report, we present an unusual case of hepatic liposarcoma of the pleomorphic subtype with literature review. In addition, we discuss theories regarding pathogenesis and the pathological and clinical features of primary hepatic liposarcoma to better outline this rare entity.Case presentationAn asymptomatic 65-year-old female was found to have a right hepatic mass on a computed tomography scan 2 years after surgical resection of the left adrenal gland and kidney for adrenocortical carcinoma. Laboratory examinations were unremarkable. Magnetic resonance imaging demonstrated a 16-mm mass in the right hepatic lobe. Adrenocortical carcinoma metastasis was suspected. Laparoscopic partial hepatectomy completely removed the tumor with clear margins. Macroscopically, the surgical specimen contained a nodular, yellow–white mass lesion 20 mm in diameter. On pathologic examination, pleomorphic, spindle-shaped tumor cells containing hypochromatic, irregularly shaped nuclei of various sizes formed fascicular structures. Scattered lipoblasts intervened in varying stages. Mitotic cells were frequent. Ki-67 labeling index was 15%. Immunohistochemically, the tumor cells were diffusely positive for vimentin and focally positive for CD34 and alpha-SMA; lipoblasts were focally positive for S-100. Tumor cells were nonreactive for SF-1, inhibin alpha, desmin, HHF35, HMB45, Melan A, MITF, c-kit, DOG1, cytokeratin AE1/AE3, h-caldesmon, STAT6, CD68, MDM2, CDK4, c17, DHEAST, 3BHSD, CD31, Factor 8, and ERG. From these findings, primary hepatic liposarcoma of pleomorphic subtype was diagnosed. The tumor recurred intrahepatically 3 years later, and the patient died 5 months after recurrence.ConclusionsIn our report, we discussed the rarity, theories regarding pathogenesis, and a review of the literature of this atypical condition. To the best of our search, this is the 14th case of primary hepatic liposarcoma and the 2nd case of the pleomorphic subtype reported throughout the world. Further research regarding the etiology of this unusual clinical entity is warranted to establish effective diagnostic and management protocols.

Identification of novel genes and pathways regulated by the orphan nuclear receptor COUP-TFII in mouse MA-10 Leydig cells†.

In males, Leydig cells are the main producers of testosterone and insulin-like 3 (INSL3), two hormones essential for sex differentiation and reproductive functions. Chicken ovalbumin upstream promoter-transcription factors I (COUP-TFI/NR2F1) and COUP-TFII (NR2F2) belong to the steroid/thyroid hormone nuclear receptor superfamily of transcription factors. In the testis, COUP-TFII is expressed and plays a role in the differentiation of cells committed to give rise to fully functional steroidogenic adult Leydig cells. Steroid production has also been shown to be diminished in COUP-TFII-depleted Leydig cells, indicating an important functional role in steroidogenesis. Until now, only a handful of target genes have been identified for COUP-TFII in Leydig cells. To provide new information into the mechanism of action of COUP-TFII in Leydig cells, we performed microarray analyses of COUP-TFII-depleted MA-10 Leydig cells. We identified 262 differentially expressed genes in COUP-TFII-depleted MA-10 cells. Many of the differentially expressed genes are known to be involved in lipid biosynthesis, lipid metabolism, male gonad development, and steroidogenesis. We validated the microarray data for a subset of the modulated genes by RT-qPCR. Downregulated genes included hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (Hsd3b1), cytochrome P450, family 11, subfamily a, polypeptide 1 (Cyp11a1), prolactin receptor (Prlr), nuclear receptor subfamily 0, group B, member 2 (Shp/Nr0b2), ferredoxin 1 (Fdx1), scavenger receptor class B, member 1 (Scarb1), inhibin alpha (Inha), and glutathione S-transferase, alpha 3 (Gsta3). Finally, analysis of the Gsta3 and Inha gene promoters showed that at least two of the downregulated genes are potentially new direct targets for COUP-TFII. These data provide new evidence that further strengthens the important nature of COUP-TFII in steroidogenesis, androgen homeostasis, cellular defense, and differentiation in mouse Leydig cells.

Pancreatic cyst dilemma: a case where genetic sequencing and immunohistochemistry impacted clinical decision making

A 78-year-old woman presented for the evaluation of an incidental pancreatic cyst. Magnetic resonance imaging (MRI) showed a 3.4-cm multiseptated cyst in the pancreatic tail with central soft tissue enhancement, suggesting a “sunburst” appearance consistent with serous cystadenoma (SCA) (A). Serum CA19-9 was normal. EUS identified a predominately solid atypical lesion with thick septations and innumerable microcysts (B). Given the modest accuracy of cross-sectional imaging and to provide reassurance to the patient, we proceeded with a fine-needle biopsy using a 19-gauge needle. Interestingly, cytopathologic analysis showed mucinous epithelium without dysplasia; therefore, a mucinous neoplasm could not be excluded (C, H&E, orig. mag. ×400). Cyst fluid/tissue sent for next-generation sequencing (PancreaSeq) was positive for a Von Hippel Lindau mutation p.C162_V165delinsF, expressed in serous cystadenomas. Given the disconcordance in results, periodic acid-Schiff (PAS) and alpha inhibin immunohistochemistry staining were performed and showed a population of PAS-positive cuboidal cells staining for inhibin, suggesting a diagnosis of SCA (D, E, Inhibin immunostain, orig. mag. ×400). Taken together, these results suggested that the mucinous epithelium was likely related to gastric mucosal contaminant from the transgastric biopsy. The final working diagnosis was SCA, supported by imaging, fluid sequencing, and immunohistochemistry. The patient was offered MRI surveillance in 6 months.

Perinatal dietary protein deficiency alters ovarian genes critical to reproductive health from one generation to another in female rat models

BackgroundSustaining a functional fertility status is highly dependent on the proper functioning of the ovary, which is a subject of normal development of ovarian follicles. Adequate and balanced nutrition forms the baseline for proper and functional reproductive makeup. This study assessed the mRNA expression level of some hormone related genes in the ovary; inhibin alpha (IHA), Estrogen receptor (ERα), Aromatase, CCAAT-enhancer binding protein alpha (CEBPA) and Fibroblast Growth Factor Receptor 1 (FGFR1) in the F1 and F2 rat generations subjected to perinatal dietary protein deficiency. Materials and methodsRats in four (4) groups were fed different grade of protein deficient diets (5%, 10%, 21% protein diets and rat chow). Histology was carried out and total RNA was extracted from ovary of the different rat groups, evaluated for quality, converted to cDNA and RT-qPCR was used to quantify levels of each gene mRNA expressed in the tissue. ResultsPresented results shows severe alteration in the expression of some of the key genes assessed, essential for the sustenance of reproductive health from one generation to another in the protein deficient group models. Inhibin alpha was downregulated across generations while CEBPA was upregulated in 5%PD groups at F1 and in 10%PD at F2, but downregulated in 5%PD group at F2, ERα was downregulated only at first generation but normalized in the second generation. Aromatase in the 10%PD group was upregulated at F1 and F2 generation, while in the 5%PD, it was downregulated only at F2. ConclusionsExposure to nutritional inadequacy at perinatal developmental stage consequentially resulted in the ovarian gene modulations that may mediate the epigenetic effects underlining the ovarian and reproductive function from one generation to another. This is reflected in the morphological study presented with ovarian degeneration and discernible ovarian cyst. The affected genes as stated in the study are very important in defining the hormonal response and overall reproductive health status of the models.

Associations of inhibin alpha beta B (INHAB) gene polymorphism with sperm quality in the population of Russian Holstein bulls

The formation and functioning of the animal reproductive system occurs as a result of the coordinated interaction of a wide range of genes. The hormone inhibin (INHA) is a marker of Sertoli cell function and exocrine testicular function (spermatogenesis state). The aim of the work was to search for associations of inhibin alpha beta B (INHAB) gene polymorphism with sperm quality in the population of Russian Holstein bulls. Sperm traits (doublet ejaculate volume, concentration, progressive motility and the total number of spermatozoa) were evaluated over three years of bull use and the highest rates for each trait were selected. 32 bulls had contrasting traits. DNA was isolated from semen samples. Samples were sequenced using the Applied Biosystems 3500 Genetic Analyzer using the BigDye® Terminator v3.1 Sequencing Standard Kit according to the protocol. Genotyping was then performed by PCR-RFLP. Data was analyzed using ANOVA. All pairwise multiple comparison between means were conducted by t-test. As a result only one nucleotide substitution A> G (rs43408735) was identified. The substitution is located in intron 1 and does not lead to a change in the amino acid composition of the protein; however, the proximity to the exon can affect the splicing processes. The frequency of the alleles in our studies was A - 0.55, G - 0.45, which differs from the population data in the ENSEMBL database, where the frequency of alleles was A - 0.31, G - 0.69. The frequency of occurrence of genotypes according to our data was AA - 0.35, AG - 0.40, GG - 0.25. Bulls with the genotype AA had a highest doublet ejaculate volume (21.00 ± 5.850 ml), total number of spermatozoa (31.43 ± 10.393 mln) than bulls with the genotypes AG (16.08±8.450 ml and 24.35 ± 14.555 mln) and GG (10.75±6.065 ml and 14.80 ± 10.613 mln) (P˂0.05). No significant differences were found in the concentration of spermatozoa and motility. But bulls with the AA genotype had the greatest concentration and motility. The bulls with GG genotype had the poorest semen quality. The study of polymorphism in the genes that determine the formation and functioning of the reproductive system will help to understand the basics of these processes, as well as use them as molecular genetic markers for culling young bulls according to sperm quality.

Genetic Analysis of Inhibin Alpha (INHα) Mutation (769G&gt;A) in Patients with Premature Ovarian Failure in a Local Population

Genetic Analysis of Inhibin Alpha (INHα) Mutation (769G&gt;A) in Patients with Premature Ovarian Failure in a Local Population

Unexpected diagnosis for a gastric polyp: Granular cell tumor: Case report and review of the literature.

A granular cell tumor (GCT) is a rare neoplasia that originates from Schwann cells. It usually appears in the skin or soft tissues, but it may occur anywhere in the body. The gastrointestinal tract is an unusual developmental site for a GCT, the esophagus being the most common site of origin for this tumor. The stomach is one of the most unique sites of origin for GCT, with less than 80 cases being mentioned in the literature. Histologically, GCTs consist of fusiform and polygonal cells, with granular cytoplasm, arranged in compact ‘nests’. Immunohistochemically, these tumors show positivity for S100 protein, CD68, CD56 and, in a smaller percentage, they are positive for other antibodies, most notably inhibin alpha. We report the case of a 52-year-old woman with a solitary GCT that had developed in the gastric cardia, discovered on a routine gastroscopy and successfully treated by endoscopic submucosal dissection.

A functional polymorphism of inhibin alpha subunit at miR-181b-1-3p-binding site regulates proliferation and apoptosis of chicken ovarian granular cells.

INHA, the gene encoding the inhibin alpha subunit, was involved in folliculogenesis in mammals, but no study was reported for its working pathway in birds. Here we hypothesize that gene polymorphism in INHA 3'UTR might influence miRNAs binding efficiency and further affect the function of this gene. Thus, we investigated the association between the 3'UTR single-nucleotide polymorphisms (SNPs) in INHA and the laying performance in chickens and further explore their possible molecular cascades in granulosa cells (GC). Five SNPs were detected in Tianfu green-shell layers and g. 22,178,975 G > A was significantly associated with total egg numbers at the age of 300days (EN, n = 286). Birds carrying the AA genotype laid more EN than those with GG (P < 0.05). The allele transition from G to A in the 3'UTR of INHA gene destroyed a binding site which was targeted by miR-181b-1-3p. The expression abundances of INHA mRNA increased firstly and then decreased with follicle growing, and reached the top in the sixth largest pre-ovulation follicle, whereas miR-181b-1-3p levels in chicken pre-hierarchical follicles had the contrary tendency. Further studies indicated that high levels of miR-181b-1-3p increased apoptosis and reduced GC proliferation while miR-181b-1-3p inhibitors decreased apoptosis and promoted GC proliferation. Additionally, depression of INHA increased apoptosis and reduced GC proliferation via a caspase-3-dependent mitochondrial pathway. Generally, the mutation in INHA 3'UTR was tightly correlated with egg production in chickens, and blocked a binding site of miR-181b-1-3p. miR-181b-1-3p inhibited GC proliferation and promoted apoptosis by targeting INHA.

34 Search for the association of polymorphisms in inhibin alpha and prolactin genes with sperm quality in Russian Holstein bulls

Abstract The formation and functioning of the animal reproductive system occurs as a result of the coordinated interaction of a wide range of genes. The hormone inhibin (INHA) is a marker of Sertoli cell function and exocrine testicular function (spermatogenesis state). The hormone prolactin (PRL) is a regulatory factor in the male reproductive system and affects gametogenesis. The aim of the work was to search for associations of inhibin alpha (INHA) and prolactin (PRL) gens polymorphism in the population of Russian Holstein bulls. Sperm traits (doublet ejaculate volume, concentration, progressive motility) were evaluated over three years of bull use and the highest rates for each trait were selected. 32 bulls had contrasting traits. DNA was isolated from semen samples. Samples were sequenced using the Applied Biosystems 3500 Genetic Analyzer using the BigDye® Terminator v3.1 Sequencing Standard Kit according to the protocol. Genotyping was then performed by PCR-RFLP. Data was analyzed using ANOVA. All pairwise multiple comparison between means were conducted by t-test. No polymorphism was detected in two regions of the PRL gene. The frequency of genotype AA in INHA gene was 0.813, the genotype AG -0.187. The GG genotype has not been identified. Bulls with the genotype AG had a higher concentration of cells (1.64 ± 0.093 billion/ml) than bulls with the genotype AA (1.35 ± 0.058) (P˂0.05). No significant differences were found in the volume of ejaculate and motility. But bulls with the AG genotype had greater ejaculate volume and motility than bulls with the AA genotype. The study of polymorphism in the genes that determine the formation and functioning of the reproductive system will help to understand the basics of these processes, as well as use them as molecular genetic markers for culling young bulls according to sperm quality. Project № АААА-А18-118021990006-9.

Effect of animal-sourced bioactive peptides on the in vitro development of mouse preantral follicles

The aim of this study was to investigate the effect of bioactive peptides (BAPT) from animal sources on the development of mouse preantral follicles in vitro. Preantral follicles were isolated and randomly divided into the following groups: an untreated group (control) and three groups supplemented with 20, 40 and 60 μg/mL BAPT, respectively. After establishing the in vitro follicle culture, the gene expression levels and hormone levels were quantified. After in vitro maturation, the developmental rates, reactive oxygen species (ROS) production levels and mitochondrial distributions of MII oocytes were investigated, followed by the analyses of embryonic developmental rates after in vitro fertilization.The results showed that BAPT promoted the growth of mouse preantral follicles. Notably, after 14 d of in vitro culture, the levels of 17 β-estradiol and progesterone were up-regulated with BAPT treatments. Moreover, the expression levels of Oct4, Bmp15, GDF9, FOXO3, Zp3, FOXL2, Inhibin alpha, SOD2, Catalase, GPx and Bcl-2 in the developing follicles were significantly up-regulated after BAPT treatments (P < 0.05), while BAPT significantly inhibited the expression levels of BAX (P < 0.05). Following BAPT treatments, the ROS production levels of MII oocytes were decreased while the mitochondrial distributions were significantly enhanced. Furthermore, increased maturation rates, fertilization and embryonic developmental rates were found in these BAPT-treated groups (P < 0.05).These results demonstrated that BAPT significantly improved the development of preantral follicles in vitro by reducing ROS-dependent cellular damages and by enhancing mitochondrial distributions, thereby promoting the further applications of animal-derived BAPT in biomedical research.

Association of inhibin alpha gene polymorphism with litter size and growth in Malabari goats of India

Abstract The present study was carried out to identify single nucleotide variations in inhibin alpha (INHA)geneto evaluate its association with production traits in Malabari goats. A total of 251 Malabari adult goats were screened for polymorphism using PCR-SSCP.Representative samples of each SSCP pattern were sequenced to ascertain the genotypes. Five fragments, 274 bp, 220 bp, 160 bp, 287 bp and 253 bp covering the two exons of INHA were analyzed. The 274 bp fragment showed two diplotypes (AA and AB) and sequencing revealed one synonymous SNP (c.129 G > A) in exon 1. The AA genotype showed significant (p ≤ 0.05) association with body length. Similar monomorphic band pattern was observed for 220 bp fragment of exon 2. The 160 bp fragment exhibited two genotypes DD and DE with a synonymous SNP c.567 G > A in exon 2. Goats carrying DD genotype had significantly (p ≤ 0.05) higher adult body weight and body length compared to DE genotype. Three diplotypes, HH, HJ and JJ with a synonymous SNP, c.651 G > A were observed at 287 bp fragment. Significantly (p C. Genotypes of INHA 253 bp (exon 2) locus had significant (p ≤ 0.05) influence on litter size, body weight and trunk circumference. The PP and PQ showed a significantly higher litter size and trunk circumference (p ≤ 0.05) compared to QQ genotypes.Present study indicated the potential role of INHA gene on reproduction and growth, hence it can be used as a candidate gene in the improvement of Malabari goats.