Steroid Production Research Articles

Kisspeptin promotes follicular development through its effects on modulation of P450 aromatase expression and steroidogenesis in sheep ovarian follicles.

The present study was conducted to ascertain whether the role of kisspeptin in promoting in vitro development of preantral follicles was through the regulation of P450 aromatase gene expression and steroidogenesis in sheep. Accordingly, the cumulus cells and oocytes were collected from different development stages of preantral follicles grown in vivo and cultured in vitro in TCM199B (Group I), TCM199B + KP (10 μg/mL) (Group II) and Standard medium + KP (10 μg/mL). To measure the steroid (Estradiol-17β; E2 and Progesterone; P4 ) synthesis through ELISA, spent culture medium was collected separately from the same in vitro groups. E2 synthesis in the spent medium collected from all the three groups showed an increasing trend from PFs' exposed to respective culture media for 3 min to 2-day culture stage but decreased thereafter till 6-day culture stage. This is followed by a sharp increase in E2 concentration in the spent medium collected after in vitro maturation. However, P4 synthesis in group III followed increased pattern as the development progressed from PFs' exposed to culture medium for 3 min to in vitro maturation stage. The steroid production was observed at all stages of in vitro development in altered supplemented conditions. The steroid synthesis in the spent medium was highest in the 6 day cultured PFs' in Standard medium + KP matured in vitro for 24 h. Therefore, supplementation of kisspeptin along with other growth factors promoted steroid production in cultured preantral follicles far better than in other media.

Novel D-modified heterocyclic androstane derivatives as potential anticancer agents: Synthesis, characterization, in vitro and in silico studies

Cancer remains a major health concern worldwide. The most frequently diagnosed types of cancer are caused by abnormal production or action of steroid hormones. In the present study, the synthesis and structural characterization of new heterocyclic androstane derivatives with D-homo lactone, 17α-(pyridine-2′′-ylmethyl) or 17(E)-(pyridine-2′′-ylmethylidene) moiety are presented. All compounds were evaluated for their anti-proliferative activity against HeLa cervical cancer cell line and non-cancerous kidney MDCK cells, where A-homo lactam compound 9A showed the greatest selectivity. Based on in vitro binding assays, N-formyl lactam compound 18 appeared to be the strong and isoform-selective ligand for ERα, while compound 9A displayed binding affinity for the GR-LBD, but also inhibited aldo-keto reductase 1C4 enzyme. Out of four selected compounds, methylpyrazolo derivative 13 showed potential for aromatase binding, while in silico studies provided insight into experimentally confirmed protein-ligand interactions.

Effects of follicle-stimulating hormone, insulin-like growth factor 1, fibroblast growth factor 2, and fibroblast growth factor 9 on sirtuins expression and histone deacetylase activity in bovine granulosa cells

Granulosa cells (GC) are critical regulators of fertility. During the process of ovarian folliculogenesis, these cells undergo profound changes while producing steroid hormones that are important to control follicular growth, oocyte maturation, and ovulation. Sirtuins are enzymes that regulate several biological processes and have been associated with control of GC function. However, how sirtuins are regulated in GC during ovarian folliculogenesis remains to be unveiled. The present study was designed to investigate effects of hormones that control GC proliferation, differentiation, and steroidogenesis on expression of the seven members of the mammalian sirtuins family (SIRT1-7) and on histone deacetylase activity of nuclear sirtuins (SIRT1, 6, and 7) in GC. Bovine granulosa cells were isolated from small antral follicles (1–5 mm) and were treated with or without follicle-stimulating hormone (FSH), insulin-like growth factor 1 (IGF-1), and fibroblast growth factors 2 (FGF2) and 9 (FGF9). Following treatments, cell proliferation was determined via a cell analyzer, estradiol synthesis and histone deacetylase activity were determined via ELISA, and sirtuins mRNA expression was determined via qPCR. Treatments with FSH and IGF-1 stimulated cell proliferation while addition of FGF2 or FGF9 suppressed estradiol production stimulated by FSH plus IGF-1. In terms of treatments that regulated sirtuins expression in GC, fibroblast growth factors were the most impactful: FGF2 alone increased SIRT1 mRNA expression in comparison to several treatments and increased mRNA abundance of SIRT2 and SIRT7 when added to the combination of FSH and IGF-1; the addition of FGF9 to the combination of FSH and IGF-1 increased mRNA expression of SIRT2, SIRT3, SIRT4, SIRT6, and SIRT7 and increased mRNA expression of SIRT5 in comparison to the negative control group that received no treatment. Also, FGF2 alone increased histone deacetylase activity of sirtuins in comparison to all treatments that contained FSH and/or IGF-1. Furthermore, several correlations were observed between treatments and sirtuins expression and activity, between estradiol or GC numbers and sirtuins expression, and between expression of sirtuins. As FGF2 and FGF9 are considered anti-differentiation factors of GC that stimulate GC proliferation while suppressing estradiol production in combination with FSH and IGF-1, data of this study suggest that sirtuins are associated with control of differentiation of bovine GC.

Landscape of PCOS co-expression gene and its role in predicting prognosis and assisting immunotherapy in endometrial cancer

BackgroundEndometrial carcinoma (EC) is the sixth most frequent malignancy in women and is often linked to high estrogen exposure. Polycystic ovarian syndrome (PCOS) is a known risk factor for EC, but the underlying mechanisms remain unclear.MethodsWe investigated shared gene signals and potential biological pathways to identify effective therapy options for PCOS- and EC-related malignancies. Weighted gene expression network analysis (WGCNA) was used to identify genes associated with PCOS and EC using gene expression data from the Gene Expression Omnibus (GEO) and Cancer Genome Atlas (TCGA) datasets. Enrichment analysis using Cluego software revealed that the steroid hormone biosynthetic process was a critical feature in both PCOS and EC. A predictive signature encompassing genes involved in steroid hormone production was developed using multivariate and least absolute shrinkage and selection operator (LASSO) regression analysis to predict the prognosis of EC. Then, we conducted further experimental verification.ResultsPatients in the TCGA cohort with high predictive scores had poorer outcomes than those with low scores. We also investigated the relationship between tumor microenvironment (TME) features and predictive risk rating and found that patients with low-risk scores had higher levels of inflammatory and inhibitory immune cells. Also, we found that immunotherapy against anti-CTLA4 and anti-PD-1/PD-L1 was successful in treating individuals with low risk. Low-risk individuals were more responsive to crizotinib therapy, according to further research performed using the “pRRophetic” R package. We further confirmed that IGF2 expression was associated with tumor cell migration, proliferation, and invasion in EC cells.ConclutionsBy uncovering the pathways and genes linking PCOS and EC, our findings may provide new therapeutic strategies for patients with PCOS-related EC.

Small-molecule Ro-08-2750 interacts with many RNA-binding proteins and elicits MUSASHI2-independent phenotypes.

RNA-binding proteins (RBPs) are key regulators of gene expression. Small molecules targeting these RBP-RNA interactions are a rapidly emerging class of therapeutics for treating a variety of diseases. Ro-08-2750 (Ro) is a small molecule identified as a competitive inhibitor of Musashi (MSI)-RNA interactions. Here, we show that multiple Ro-dependent cellular phenotypes, specifically adrenocortical steroid production and cell viability, are Musashi-2 (MSI2)-independent. Using an unbiased proteome-wide approach, we discovered Ro broadly interacts with RBPs, many containing RRM domains. To confirm this finding, we leveraged the large-scale ENCODE data to identify a subset of RBPs whose depletion phenocopies Ro inhibition, indicating Ro is a promiscuous inhibitor of multiple RBPs. Consistent with broad disruption of ribonucleoprotein complexes, Ro treatment leads to stress granule formation. This strategy represents a generalizable framework for validating the specificity and identifying targets of RBP inhibitors in a cellular context.

P-394 Neurotrophin-4 supplementation during human secondary follicle in-vitro-culture supports morphologically normal blastocyst formation

Abstract Study question Can matrix-free culture system supplemented with Neurotrophic factor 4 (NT4) improve human follicular development and achieve mature and fertile oocytes in vitro? Summary answer NT4 promotes the growth, steroid hormone production of human secondary follicles cultured in vitro (including post- and pre-pubertal patients), thereby yielding mature and fertile oocytes. What is known already Reconstituting folliculogenesis in vitro play vital roles in reproductive biology research, fertility preservation and reproductive toxicity testing. However, fertile oocyte from in-vitro-grown (IVG) human follicle remains unachieved. It has been demonstrated that NT4 promotes in-vitro survival, growth, and maturation of mice secondary follicles. NT4 and its receptors have been detected in human ovaries. However, if the advance proved in mice model is effective to replicate in human has not been reported. Study design, size, duration Discarded ovarian tissues after ovarian tissue cryopreservation (OTC) were collected from 6 patients aged from 11 to 21 years old who underwent unilateral oophorectomy for fertility preservation, after obtaining written informed consent. Isolated secondary follicles were cultured in vitro with or without NT4 in a matrix-free system for 4-6 weeks individually. Participants/materials, setting, methods Secondary follicles isolated from each patient were randomly assigned to control or NT4 group, followed by incubation on the ultra-low attachment microplate individually. Follicle growth and survival were assessed by microscopy. Anti-Müllerian hormone (AMH), estradiol and progesterone levels were quantified in the medium. Oocyte marker expression was evaluated by DEAD box polypeptide 4 (Ddx4) staining. Oocyte mature and fertile potential were assessed by in-vitro maturation and intracytoplasmic sperm injection (ICSI) with donated sperm, separately. Main results and the role of chance In control condition, isolated follicles survived for 4-6 weeks with increased diameters over time (P < 0.05), reaching a terminal diameter of 967.00 ± 41.1 μm with the confirmed induction of steroidogenesis and expression of widely accepted oocyte markers (DDX4). Three out of twelve (25%) alive follicles were matured successfully in vitro, most of which produced morphologically normal MII oocytes with sizes of 120 ± 2.1 μm. The diameter and steroid hormone production were significantly higher in the group cultured with NT4 than in the control group (P < 0.05). An increased efficiency of MII oocyte production in the NT4 group was also observed, while the difference was not statistically significant. MII oocyte obtained from the control group showed abnormal fertilization after ICSI. In contrast, MII oocyte acquired from the NT4 group progressed to available blastocyst. Limitations, reasons for caution The population included were all patients with thalassemia. Whether this culture system is effective for patients with other diseases is unknown. Limited by the small sample, the effect of NT4 on maturation competence needs further confirmation. Oocytes obtained have not been quantified with ploidy status and epigenetic signatures. Wider implications of the findings Surplus ovarian tissue after OTC, which is otherwise discarded, may serve as an additional precious source of fertile oocytes for fertility preservation, even for pre-pubertal girls, without a threat of tumor reintroduction. The system described here will provide a powerful research tool for reproductive biology research and reproductive toxicity testing. Trial registration number not applicable

Progress of 3-ketosteroid Δ1-dehydrogenases for steroid production

Progress of 3-ketosteroid Δ1-dehydrogenases for steroid production

Putative adverse outcome pathway development based on physiological responses of female fathead minnows to model estrogen versus androgen receptor agonists

Several adverse outcome pathways (AOPs) have linked molecular initiating events like aromatase inhibition, androgen receptor (AR) agonism, and estrogen receptor (ER) antagonism to reproductive impairment in adult fish. Estrogen receptor agonists can also cause adverse reproductive effects, however, the early key events (KEs) in an AOP leading to this are mostly unknown. The primary aim of this study was to develop hypotheses regarding the potential mechanisms through which exposure to ER agonists might lead to reproductive impairment in female fish. Mature fathead minnows were exposed to 1 or 10 ng 17α-ethynylestradiol (EE2)/L or 10 or 100 µg bisphenol A (BPA)/L for 14 d. The response to EE2 and BPA was contrasted with the effects of 500 ng/L of 17β-trenbolone (TRB), an AR agonist, as well as TRB combined with the low and high concentrations of EE2 or BPA tested individually. Exposure to 10 ng EE2/L, 100 µg BPA/L, TRB, or the various mixtures with TRB caused significant decreases in plasma concentrations of 17β-estradiol. Exposure to TRB alone caused a significant reduction in plasma vitellogenin (VTG), but VTG was unaffected or even increased in females exposed to EE2 or BPA alone or, in most cases, in mixtures with TRB. Over the course of the 14-d exposure, the only treatments that clearly did not affect egg production were 1 ng EE2/L and 10 µg BPA/L. Based on these results and knowledge of hypothalamic–pituitary–gonadal axis function, we hypothesize an AOP whereby decreased production of maturation-inducing steroid leading to impaired oocyte maturation and ovulation, possibly due to negative feedback or direct inhibitory effects of membrane ER activation, could be responsible for causing adverse reproductive impacts in female fish exposed to ER agonists.

Driving the conversion of phytosterol to 9α-hydroxy-4-androstene-3,17-dione in Mycolicibacterium neoaurum by engineering the supply and regeneration of flavin adenine dinucleotide

BackgroundThe conversion of phytosterols to steroid synthons by engineered Mycolicibacteria comprises one of the core steps in the commercial production of steroid hormones. This is a complex oxidative catabolic process, and taking the production of androstenones as example, it requires about 10 equivalent flavin adenine dinucleotide (FAD). As the high demand for FAD, the insufficient supply of FAD may be a common issue limiting the conversion process.ResultsWe substantiated, using the production of 9α-hydroxy-4-androstene-3,17-dione (9-OHAD) as a model, that increasing intracellular FAD supply could effectively increase the conversion of phytosterols into 9-OHAD. Overexpressing ribB and ribC, two key genes involving in FAD synthesis, could significantly enhance the amount of intracellular FAD by 167.4% and the production of 9-OHAD by 25.6%. Subsequently, styrene monooxygenase NfStyA2B from Nocardia farcinica was employed to promote the cyclic regeneration of FAD by coupling the oxidation of nicotinamide adenine dinucleotide (NADH) to NAD+, and the production of 9-OHAD was further enhanced by 9.4%. However, the viable cell numbers decreased by 20.1%, which was attributed to sharply increased levels of H2O2 because of the regeneration of FAD from FADH2. Thus, we tried to resolve the conflict between FAD regeneration and cell growth by the overexpression of catalase and promotor replacement. Finally, a robust strain NF-P2 was obtained, which could produce 9.02 g/L 9-OHAD after adding 15 g/L phytosterols with productivity of 0.075 g/(L h), which was 66.7% higher than that produced by the original strain.ConclusionsThis study highlighted that the cofactor engineering, including the supply and recycling of FAD and NAD+ in Mycolicibacterium, should be adopted as a parallel strategy with pathway engineering to improve the productivity of the industrial strains in the conversion of phytosterols into steroid synthons.

Time- and glucose-dependent differentiation of 3T3-L1 adipocytes mimics dysfunctional adiposity

The 3T3-L1 murine adipocyte cell line remains one of the most widely used models to study the mechanisms of obesity and related pathologies. Most studies investigate such mechanisms using mature adipocytes that have been chemically induced to differentiate for 7 days in media containing 25 mM glucose. However, the dysfunctional characteristics commonly observed in obesity including adipocyte hypertrophy, increased expression of inflammatory markers, enhanced production of reactive oxygen species (ROS), increased steroidogenic enzyme expression/activity and production of steroid hormones, are not necessarily mimicked in these cells. The aim of this study was to provide an inexpensive model which represents the well-known characteristics of obesity by manipulating the time of adipocyte differentiation and increasing the concentration of glucose in the cell media. Our results showed a glucose- and time-dependent increase in adipocyte hypertrophy, ROS production and gene expression of the pro-inflammatory cytokine interleukin-6 (IL-6), as well as a time-dependent increase in lipolysis and in the gene expression of the chemokine monocyte chemoattractant protein 1 (MCP1). We also showed that gene expression of the steroidogenic enzymes 11-beta-hydroxysteroid dehydrogenase type 1 (11βHSD1), 17βHSD type 7 and 12, as well as CYP19A1 (aromatase), were significantly higher in the hypertrophic model relative to the control adipocytes differentiated using the conventional method. The increase in 11βHSD1 and 17βHSD12 expression was consistent with the enhanced conversion of cortisone and androstenedione to cortisol and testosterone, respectively. As these characteristics reflect those commonly observed in obesity, hypertrophic 3T3-L1 adipocytes are an appropriate in vitro model to study mechanisms of adipocyte dysfunction in an era where the rise in obesity incidence is a global health concern, and where access to adipose tissue from obese patients are limited.

Effect of anti-GnRH vaccine on testicular tissues in stallions

Anti-GnRH Vaccine is known as an alternative to surgical castration. The main advantage is the reversibility of immunization in 90% of the cases (Bruyas et al. Reprod Dom Anim, p.55: https://doi.org/10.1111/rda.12800). Anti-GnRH immunization reduces fertility by inhibition of gonadotropin synthesis involved in steroid production and spermatogenesis. Only few papers report information on the recovery of sperm production after prime immunization (twice 4 weeks apart) without booster in some stallions. We aimed to assess the effect of the vaccination protocol (Primo immunization or/and boosters) and the delay between the last injection of anti-GnRH vaccine and surgical castration on testicular histology. Twelve unvaccinated (control) and 28 immunized Lusitanian stallions were castrated at different time points after the last immunization (range 0.5-14 months). Vaccination protocol was a primo vaccination by intramuscular injection of 1 mL IMPROVAC® twice 4 weeks apart, followed by booster(s) when stallion behavior returned or scrotal size re-increased (range 1-10 injections). Testes were fixed in formaldehyde (10%) and stained by HES (Hematoxylin-Eosin-Saffron) and Androgen Receptor immunohistochemistry. Testicular size and spermatogenesis were evaluated by 7 parameters: Johnsen scores, seminiferous tubule diameter, number and density of Sertoli cells, number and density of germ cells, and the ratio of germ to Sertoli cells.Statistical analysis was performed by ANOVA. Values are given as mean ±standard deviation. Testis size was larger for unvaccinated controls (88±5 mm; P<0.001) compared to vaccinated stallions (70 ±8 mm). The Johnsen scores were higher in control (9.3 ±0.3; P<0.0001) compared to vaccinated stallions (8.0 ±1.2). Seminiferous tubule diameters, number and density of germ cells, and the ratio of germ to Sertoli cells were larger in control stallions: respectively 173 ±10 µm (P<0.001), 180 ±34 germ cells per tubule (P<0.001), 6330 ±897 germ cells per mm² of tubule (P<0.001), and 8.2 ±1.5 (P<0.001) germ cells per Sertoli cell compared to vaccinated stallions: 135 ±22 µm, 83 ±47 germ cells per tubule, 4404 ±1454 germ cells per mm² of tubule, and 3.3 ±1.9 germ cells per Sertoli cell. Conversely, Sertoli cell numbers and density were larger in vaccinated: 27 ±4 Sertoli cells per tubule (P=0.005) and 1720 ±582 Sertoli cells per mm² of tubule (P<0.001) compared to 23 ±4 Sertoli cells per tubule and 817 ±165 Sertoli cells per mm² of tubule in controls. There was no significant difference among the vaccination protocols (e.g., number of boosters, delay between last injection and gonadectomy) on these parameters. In conclusion, multiple rounds of anti-GnRH immunization in stallions: (1) induced testicular hypoplasia is more dependent on stallion rather the vaccination protocol, and (2) had no reversible effects on testes (according to Johnsen scores).

An in vitro model for equine placental research

A fundamental understanding of placental development and its associated pathologies is crucial to mitigating the losses associated with placental disorders. Organoids are widely used to investigate the developmental biology of organs and in disease modeling. In humans, placental organoids have been successfully developed using trophoblast cells. In this study, we aimed to 1) develop placental organoids using pre-term and term equine placentae, 2) evaluate the organoid structures, 3) compare the organoid transcriptome with their respective chorioallantois (CA), and 4) evaluate organoids based on their starting material (pre-term vs term). The CA was collected from healthy placentae following elective abortion (80-100d of gestation) and following spontaneous term parturition. CA samples were processed for culture, fixed for histology, and stored in RNAlater. For culture, each CA sample was enzymatically digested to a single cell suspension and was seeded into 50µL Matrigel Matrix droplets surrounded by medium. Passage-3 organoids (P3) were used for histology (H&E), immunohistochemistry (IHC), and transcriptomic analysis. Steroid production and conversion were measured by mass spectrometry (MS) of culture media. After optimization of culture conditions, we produced 3D-organoids (Pre-Term, n=6 and Term, n=3). Organoids were passaged up to 12 times over ±3 months and were successfully frozen and re-cultured post-thaw. Total extracted RNA from initial CA samples and organoids was sequenced (150PE; ∼43 million reads/sample). The genes expressed in the organoidsclosely resemble those of the tissue of origin, 19.4% of the expressed genes were differentially expressed (DEGs) between the initial CA and P3 (n=3,949 genes; FDR<0.001). H&E staining of both CA and the organoids indicates the epithelial origin of the cells within the organoids. The enriched expression of trophoblast-specific genes PLAC8B, GATA3, KRT7, VGLL1, and EGFR, and the presence of the trophoblast-specific genes TFAP2A and TFAP2C, and GATA2 in the organoids confirm their trophoblast identity. The presence of SOX2 and OCT4, as demonstrated by IHC, confirms the pluripotency of the organoid cells. The SRD5A1 and SRD5A3 genes were expressed in all organoids and our preliminary MS analysis demonstrated the conversion of progesterone to 5α-DHP, further suggesting functional integrity. Comparison between Pre-Term and Term organoids revealed 552 DEGs (FDR<0.001), with downregulation of immune/inflammatory/defense responses and blood vessel development, and upregulation of molecule transport, development, and adhesion in the Pre-Term group. To conclude, this is the first report of placental organoids in domestic animals. The organoids show structural, transcriptomic, and functional similarities to the equine placenta in vivo and consequently could be used as an in vitro model to study placental development and associated pathologies. Funded by The Foundation for The Horse, GJCRF, UGent BOF, BAEF, L'Oréal-UNESCO FWO, and the Center for Equine Health at UC Davis.

The inhibition of steroid hormones determines the fate of IPC-366 tumor cells, highlighting the crucial role of androgen production in tumor processes

Inflammatory mammary cancer (IMC) is a disease that affects female dogs. It is characterized by poor treatment options and no efficient targets. However, anti-androgenic and anti-estrogenic therapies could be effective because IMC has a great endocrine influence, affecting tumor progression. IPC-366 is a triple negative IMC cell line that has been postulated as a useful model to study this disease. Therefore, the aim of this study was to inhibit steroid hormones production at different points of the steroid pathway in order to determine its effect in cell viability and migration in vitro and tumor growth in vivo. For this purpose, Dutasteride (anti-5αReductase), Anastrozole (anti-aromatase) and ASP9521 (anti-17βHSD) and their combinations have been used. Results revealed that this cell line is positive to estrogen receptor β (ERβ) and androgen receptor (AR) and endocrine therapies reduce cell viability. Our results enforced the hypothesis that estrogens promote cell viability and migration in vitro due to the function of E1SO4 as an estrogen reservoir for E2 production that promotes the IMC cells proliferation. Also, an increase in androgen secretion was associated with a reduction in cell viability. Finally, in vivo assays showed large tumor reduction. Hormone assays determined that high estrogen levels and the reduction of androgen levels promote tumor growth in Balb/SCID IMC mice. In conclusion, estrogen levels reduction may be associated with a good prognosis. Also, activation of AR by increasing androgen production could result in effective therapy for IMC because their anti-proliferative effect.

Autophagy regulates sex steroid hormone synthesis through lysosomal degradation of lipid droplets in human ovary and testis

Autophagy is an evolutionarily conserved process that aims to maintain the energy homeostasis of the cell by recycling long-lived proteins and organelles. Previous studies documented the role of autophagy in sex steroid hormone biosynthesis in different animal models and human testis. Here we demonstrate in this study that sex steroid hormones estrogen and progesterone are produced through the same autophagy-mediated mechanism in the human ovary in addition to the human testis. In brief, pharmacological inhibition and genetic interruption of autophagy through silencing of autophagy genes (Beclin1 and ATG5) via siRNA and shRNA technologies significantly reduced basal and gonadotropin-stimulated estradiol (E2), progesterone (P4) and testosterone (T) production in the ex vivo explant tissue culture of ovary and testis and primary and immortalized granulosa cells. Consistent with the findings of the previous works, we observed that lipophagy, a special form of autophagy, mediates the association of the lipid droplets (LD)s with lysosome to deliver the lipid cargo within the LDs to lysosomes for degradation in order to release free cholesterol required for steroid synthesis. Gonadotropin hormones are likely to augment the production of sex steroid hormones by upregulating the expression of autophagy genes, accelerating autophagic flux and promoting the association of LDs with autophagosome and lysosome. Moreover, we detected some aberrations at different steps of lipophagy-mediated P4 production in the luteinized GCs of women with defective ovarian luteal function. The progression of autophagy and the fusion of the LDs with lysosome are markedly defective, along with reduced P4 production in these patients. Our data, together with the findings of the previous works, may have significant clinical implications by opening a new avenue in understanding and treatment of a wide range of diseases, from reproductive disorders to sex steroid-producing neoplasms, sex steroid-dependent malignancies (breast, endometrium, prostate) and benign disorders (endometriosis).

Ketoconazole: A Promising Drug for Fungal Infections

Ketoconazole is a synthetic imidazole antifungal agent that can be used to treat surface fungal infections, genital, chronic mucocutaneous candidosis, and candidosis. Ketoconazole is a synthetic imidazole derivative that is effective after oral administration, unlike older imidazole antifungals like miconazole and clotrimazole. It works well for fungal infections on the skin, such as dermatophytosis and genital candidiasis. It has become useful for the treatment of persistent mucocutaneous candidiasis. In immunocompromised individuals, ketoconazole is also used to prevent fungal infections. It has lately been utilised as a therapy for advanced prostatic cancer because it inhibits the production of adrenal sex hormone. Although oral ketoconazole has been subjected to labelling modifications and market withdrawal because of substantial adverse effects, topical ketoconazole is widely regarded to be efficacious, safe, and best for treating superficial fungal infections. Onychomycosis, blepharitis, and hair loss are some of the new dermatologic indications for topical ketoconazole usage.

Comparative developmental genomics of sex-biased gene expression in early embryogenesis across mammals

BackgroundMammalian gonadal sex is determined by the presence or absence of a Y chromosome and the subsequent production of sex hormones contributes to secondary sexual differentiation. However, sex chromosome-linked genes encoding dosage-sensitive transcription and epigenetic factors are expressed well before gonad formation and have the potential to establish sex-biased expression that persists beyond the appearance of gonadal hormones. Here, we apply a comparative bioinformatics analysis on a pair of published single-cell datasets from mouse and human during very early embryogenesis—from two-cell to pre-implantation stages—to characterize sex-specific signals and to assess the degree of conservation among early acting sex-specific genes and pathways.ResultsClustering and regression analyses of gene expression across samples reveal that sex initially plays a significant role in overall gene expression patterns at the earliest stages of embryogenesis which potentially may be the byproduct of signals from male and female gametes during fertilization. Although these transcriptional sex effects rapidly diminish, sex-biased genes appear to form sex-specific protein–protein interaction networks across pre-implantation stages in both mammals providing evidence that sex-biased expression of epigenetic enzymes may establish sex-specific patterns that persist beyond pre-implantation. Non-negative matrix factorization (NMF) on male and female transcriptomes generated clusters of genes with similar expression patterns across sex and developmental stages, including post-fertilization, epigenetic, and pre-implantation ontologies conserved between mouse and human. While the fraction of sex-differentially expressed genes (sexDEGs) in early embryonic stages is similar and functional ontologies are conserved, the genes involved are generally different in mouse and human.ConclusionsThis comparative study uncovers much earlier than expected sex-specific signals in mouse and human embryos that pre-date hormonal signaling from the gonads. These early signals are diverged with respect to orthologs yet conserved in terms of function with important implications in the use of genetic models for sex-specific disease.

Processive kinetics in the three-step lanosterol 14α-demethylation reaction catalyzed by human cytochrome P450 51A1

Cytochrome P450 (P450, CYP) family 51 enzymes catalyze the 14α-demethylation of sterols, leading to critical products used for membranes and the production of steroids, as well as signaling molecules. In mammals, P450 51 catalyzes the 3-step, 6-electron oxidation of lanosterol to form (4β,5α)-4,4-dimethyl-cholestra-8,14,24-trien-3-ol (FF-MAS). P450 51A1 can also use24,25-dihydrolanosterol (a natural substrate in the Kandutsch-Russell cholesterol pathway). 24,25-Dihydrolanosterol and the corresponding P450 51A1 reaction intermediates, the 14α-alcohol and -aldehyde derivatives of dihydrolanosterol, were synthesized to study the kinetic processivity of the overall 14α-demethylation reaction of human P450 51A1. A combination of steady-state kinetic parameters, steady-state binding constants, dissociation rates of P450-sterol complexes, and kinetic modeling of the time course of oxidation of a P450-dihydrolanosterol complex showed that the overall reaction is highly processive, withkoffrates of P450 51A1-dihydrolanosterol and the 14α-alcohol and 14α-aldehyde complexes being 1 to 2 orders of magnitude less than the forward rates of competing oxidations. epi-Dihydrolanosterol (the 3α-hydroxy analog) was as efficient as the common 3β-hydroxy isomer in the binding and formation of dihydro FF-MAS. The common lanosterol contaminant dihydroagnosterol was found to be a substrate of human P450 51A1, with roughly one-half the activity of dihydrolanosterol. Steady-state experiments with 14α-methyl deuterated dihydrolanosterol showed no kinetic isotope effect, indicating that C-14αC-H bond breaking is not rate-limiting in any of the individual steps. The high processivity of this reaction generates higher efficiency and also renders the reaction less sensitive to inhibitors.

Stimulatory effects of vasopressin on progesterone production and BMP signaling by ovarian granulosa cells

The aim of the present study was to clarify the effects of arginine vasopressin (AVP) on ovarian steroid production and its functional relationship to the ovarian bone morphogenetic protein (BMP) system. The results showed that AVP treatment significantly increased gonadotropin- and forskolin-induced progesterone synthesis by primary culture of rat granulosa cells and human granulosa cells, respectively. In contrast, estradiol production was not significantly affected by AVP. Treatment with AVP significantly increased forskolin-induced cAMP synthesis by human granulosa cells and mRNA levels of the progesterogenic enzymes CYP11A1 and HSD3B2 in the cells. On the other hand, AVP also enhanced BMP-15-induced phosphorylation of SMAD1/5/9 and ID1 transcription. It was further revealed that the expression levels of BMP receptors, including ALK3, ALK6 and BMPR2, were upregulated by AVP. Collectively, the results indicate that AVP stimulates progesterone production via the cAMP-PKA pathway with upregulation of BMP signaling that inhibits progesterone production, which may lead to fine adjustment of progesterone biosynthesis by granulosa cells.

The puzzle of extensive tinea incognito with a young woman: a case report a tinea incognito with Cushing’s disease

Introduction: Tinea incognito is a rare condition commonly found in abused topical or systemic steroids. Considering their immunosuppression status, Cushing’s syndrome predisposes patients to infectious diseases. Most patients of Cushing’s syndrome are as a result of adrenocorticotropic hormone production from a pituitary adenoma, which is referred to as Cushing’s disease. Here, we reported a rare co-occurrence of tinea incognito and Cushing’s syndrome secondary to a giant pituitary tumor. Case presentation: Extensive cutaneous and nail lesions were diagnosed with the tinea incognito and tinea unguium, and endocrine or rheumatic disorders were considered as underlying disease-causing extensive tinea incognito. A series of laboratory and imaging tests were performed. We found that a giant pituitary tumor was the primary etiology of multiple cutaneous appearances and secondary disorders. There was an exceptionally unusual co-occurrence of extensive tinea incognito and pituitary adenoma in our case. Discussion: We observed a rare co-occurrence of extensive tinea incognito and pituitary adenoma, which should be taken into consideration in the urgent diagnostic criterion of tinea incognito given the potential underlying condition. Conclusion: Tinea incognito is not only associated with a history of topical or systemic steroid use, but also the association that underlying diseases related to excessive endogenous steroid production are supported to be taken into account.

The Relationship Between Menstrual Cycle Disorders In Obese Adolescents

Menstrual cycle disorders are the main symptom of anovulation, a phenomenon accompanied by decreased ovarian steroid secretion and production. Weight gain and an increase in adipose tissue in obese women, especially in the midsection, can disrupt the balance of steroid hormones such as androgens, estrogens, and sex hormone-binding globulins, which can affect the menstrual cycle in adolescents. The mechanism of obesity that is most often associated with menstrual cycle disorders. On that basis, this study aims to assess the relationship between insulin resistance and menstrual cycle disorders in obese adolescents. Method : This study used a cross-sectional design with comparisons where in this study the researchers compared the levels of the Homeostatic Model obese adolescent women who had normal menstrual cycles and abnormal menstrual cycles. Result : the group with abnormal menstrual cycles were found to be lower than those in the normal menstrual cycle group, but no significant difference was found. In addition, the abnormal menstrual cycle group were found to be lower but not significant. Conclusion : Obesity no significant relationship to the menstrual cycle in obese adolescent girls