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Background/aim: Biomaterials are essential in modern medicine, both for patients and research. Their ability to acquire and maintain functional vascularization is currently debated. The aim of this study was to evaluate the vascularization induced by two collagen-based scaffolds (with 2D and 3D structures) and one non-collagen scaffold implanted on the chick embryo chorioallantoic membrane (CAM)., Materials and Methods: Classical stereomicroscopic image vascular assessment was enhanced with the IKOSA software by using two applications: the CAM assay and the Network Formation Assay, evaluating the vessel branching potential, vascular area, as well as tube length and thickness., Results: Both collagen-based scaffolds induced non-inflammatory angiogenesis, but the non-collagen scaffold induced a massive inflammation followed by inflammatory-related angiogenesis. Vessels branching points/Region of Interest (Px^2) and Vessel branching points/Vessel total area (Px^2), increased exponentially until day 5 of the experiment certifying a sustained and continuous angiogenic process induced by 3D collagen scaffolds., Conclusion: Collagen-based scaffolds may be more suitable for neovascularization compared to non-collagen scaffolds. The present study demonstrates the potential of the CAM model in combination with AI-based software for the evaluation of vascularization in biomaterials. This approach could help to reduce and replace animal experimentation in the pre-screening of biomaterials., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: A real challenge for patients with rheumatoid arthritis (RA) and rheumatologists is primary nonresponse status (PNRS) or secondary nonresponse status (SNRS) to various therapies. Despite their detrimental influence on patient life quality, PNRS and SNRS have no accurate definition and no early predictive criteria for their development exist. Patients with RA under 40 years of age are rare, hence PNRS and SNRS data for this age group are scarce. This study examined the PNRS and SNRS according to sex, age, BMI, therapy type, and duration., Patients and Methods: Retrospectively, 115 patients with RA having PNRS and/or SNRS were stratified by age (22-39, 40-59, and 60-81). The association between body mass index (BMI), proinflammatory cytokines inhibitors, JAK inhibitors, and TNF-alpha inhibitors, sex, age, and PNRS and SNRS was examined., Results: All three proinflammatory cytokine inhibitors (rituximab, tocilizumab, and abatacept) were associated with PNRS and SNRS in women with a high BMI aged 40-59 years. Abatacept-related PNRS and SNRS was significant in women with normal BMI aged 60-81 years. Adalimumab, infliximab, and golimumab affected SNRS differently in women with normal BMI aged 22-39 years and women with high BMI aged 60-81 years. Etanercept and infliximab were associated with SNRS status in men with high-BMI aged 40-59 years., Conclusion: PNRS and SNRS development in patients with RA is significantly influenced by age, sex, and BMI, but most importantly is closely and differentially related to therapy type and duration., (Copyright © 2023, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: Colon cancer liver metastases with desmoplastic growth pattern (dGP) have a highly heterogeneous therapy response. The aim of the study was to evaluate the dGP liver metastasis molecular profile from a chemo-naive patient by mimicking metastatic process on an experimental chick embryo chorioallantoic membrane (CAM) model., Materials and Methods: Three successive CAM passages of dGP human colorectal liver metastases were immunophenotyped for keratin (K) 8, and 20, CLIC1, VEGF, EGFR, CD34, podoplanin, Ki67, E-cadherin and vimentin., Results: Metastatic cells gradually lost K20 while K8, E-cadherin and vimentin heterogeneously increased during passages. VEGF, CLIC 1, EGFR expression increased in metastatic cells especially at the tumor graft periphery. Scattered proliferating and non-proliferating podoplanin-positive tumor cells, lymphatic and blood vessels were heterogeneously detected in tumor xenografts depending on passage stage., Conclusion: By mimicking repetitive metastatic processes we proved that metastatic cells change their phenotype. This may explain why not all metastases have a similar response to therapy., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: Chloride intracellular channel protein 1 (CLIC1) activates inflammasomes in rheumatoid (RA) and psoriatic (PsA) arthritis. We studied CLIC1 expression in RA and PsA patients' skin with vasculitis and its variability depending on the therapy used., Materials and Methods: CLIC1 immunoexpression was evaluated in the vascular (CLIC1-V) and stromal (CLIC1-S) compartments of the RA and PsA skin biopsies of patients treated with methotrexate (MTX), leflunomid (LFN), corticotherapy (CT), or biological therapies., Results: MTX significantly reduced CLIC1-S expression (p=0.016), whereas LFN decreased CLIC1-V (p<0.001). LFN therapy duration also correlated with CLIC1-V (p<0.001). CT decreased CLIC1-S expression (p=0.006). CLIC1-S expression persisted in skin biopsies despite of erythrocyte sedimentation rate (ESR, p=0.018) and C reactive protein (CRP, p=0.0026) normalisation. For PsA, CLIC1-S expression significantly related to MTX (p<0.022). Both CLIC1-S (p<0.001) and CLIC1-V (p=0.007) decreased by biological therapies in RA., Conclusion: CLIC1 expression is strongly influenced by the therapy used. Our data strongly support the extensive evaluation of CLIC1 in RA as a potential marker of inflammation and tool to predict therapy response., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: Angiogenic growth factors expression is not known in the normal cornea. The aim was to study corneal gene expression profile of VEGF and PDGF pathways influencing the avascular state of cornea., Materials and Methods: cDNA synthesis was performed from mRNA extracted from five fresh pig corneas followed by cDNA synthesis and analysis of VEGF and PDGF pathways by TaqMan Array gene expression profile., Results: Normal pig cornea lacks VEGFR2 and VEGFR3 gene expression. MK2 and AKT1 genes were significantly overexpressed (p=0.000684, p=0.050995, respectively). Six PDGF pathway genes were overexpressed: TIAM1 (p=0.047), PIK3CA (p=0.00005), IKBKG (p=0.000006), PAK4 (p=0.034), RAC1 (p=0.000006 and PTGS2, p=0.00375). PDGF A was up-regulated, but not with a statistical significance (p=0.79911), while PDGFRα was down-regulated and PDGFRβ was not expressed., Conclusion: Normal cornea avascularity is given by growth factor receptors down-regulation. Rapid corneal neovascularisation is induced by activation of the main angiogenic growth factors that induce angiogenic cascade and vessel recruitment., (Copyright© 2021, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: The role of podoplanin (PDPN) and homebox prospero gene 1 (PROX1) in early stages of pancreatic islet changes induced by hypercaloric diet is unclear. The aim of this study was to study PDPN and PROX1 variability in pancreatic islets after a hypercaloric diet in a rat experimental model., Materials and Methods: Pancreatic biopsies harvested from Sprague-Dawley rats at 3, 6, and 9 weeks following hypercaloric diet intake were evaluated for morphological and molecular changes of Langerhans islets based on PDPN and PROX1 expression Results: Six weeks of hypercaloric diet induced hypertrophy of pancreatic islets with focal expression of Pdpn and Prox1 mRNA. At 9 weeks of hypercaloric diet, strong peri-insular inflammation was found around hypertrophic islets highly expressing PDPN, and lacking Prox1 mRNA and protein expression., Conclusion: This is the first report of Pdpn and Prox1 mRNA expression variability and involvement in early steps of pancreatic islet changes following hypercaloric food intake., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: Mast cells (MCs) represent the most controversial non-malignant element of the tumor microenvironment. Our aim was to study how MCs density and distribution (intratumoral-MCit versus peritumoral-MCpt) relate to tumor grade and molecular subtypes., Materials and Methods: MCs tryptase immunohistochemistry was performed on 80 cases of breast carcinomas., Results: For Luminal A tumors, a partial correlation was detected between MCit and progesterone receptor (PR) (p=0.005). Luminal B tumors showed a significant correlation between MCpt and age (p=0.009), estrogen receptor (ER) (p=0.017) and PR (p=0.035). MCit and MCpt were strongly interrelated in this subtype (p=0.002) and in triple-negative breast cancers (p=0.002). In HER2 subtype, MCpt tumors were significantly correlated with HER2 (p=0.044). In G2 tumors, MCpt correlated with ER (p=0.015) and PR (p=0.038) while in G3 tumors ER correlated with both MCit (p=0.009) and MCpt (p=0.000487) tumors., Conclusion: MCs dynamics are strongly influenced by hormone receptors and HER2 status. MCit increased in aggressive tumor types and is a worse prognostic factor., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: The aim of this study was to evaluate SOX2 expression in pituitary adenomas and its correlation to their secretory state and clinicopathological parameters., Patients and Methods: Thirty-four patients were clinically evaluated and surgery was recommended for tumor removal. Histopathological diagnosis by hematoxylin eosin staining was followed by immunohistochemistry for pituitary hormones and SOX2 co-expression., Results: Fourteen of the 34 cases were GH-secreting adenomas, 10 were prolactinomas and 10 non-functioning pituitary adenomas. SOX2-positive expression was detected in 47.05% of total cases: 8 GH-secreting adenomas (57.14%), 6 prolactinomas (60%) and 2 non-functioning adenomas (20%). SOX2 positivity was significantly higher amongst secreting adenomas (p=0.041). SOX2-negative tumors were significantly associated with corticotrophin deficiency (p=0.047) and gonadotrophin deficiency (p=0.041). No correlation with tumor size or extrasellar extension was detected., Conclusion: SOX2 is differentially expressed in pituitary adenomas and influences the secretory state or clinical behavior of pituitary adenomas., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Aim To assess the onset of early left ventricular (LV) systolic and diastolic function impairment and the subclinical atherosclerosis following chemotherapy in patients diagnosed with acute myeloid leukemia (AML)., Materials and Methods: Thirty patients diagnosed with AML with no cardiac history, having LV ejection fraction (LVEF) >50%, were evaluated at baseline and 6 months after starting four cycles of chemotherapy. We measured LV function, global longitudinal strain and subclinical atherosclerosis markers: intima-media thickness (IMT), arterial stiffness aortic pulse wave velocity (PWVAo) and ankle-brachial index (ABI)., Results: LVEF had decreased at 6 months after treatment initialization (p<0.001), the same changes being observed for LV fraction shortening (p<0.001), mitral annular plane systolic excursion and S' wave (p<0.001 and p<0.05). Bilateral IMT and PWVAo significantly increased, 12 out of 30 patients (40%) had LVEF ≤50% after 6 months of chemotherapy, five of them receiving daunorubicin at more than 500 mg/m 2 /injection., Conclusion: LV function is impaired after 6 months of chemotherapy, with early changes of subclinical atherosclerosis becoming evident., (Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Aim: To characterize baby hamster kidney fibroblast (BHK 21/C13) cells and test the effects of antibodies against podoplanin and disodium cromolyn on BHK 21/C13 cell line-derived tumors grown on chick embryo chorioallantoic membrane (CAM)., Material and Methods: BHK 21/C13 cell-derived fibrosarcomas developed in hamsters were implanted on CAM and treated with anti-podoplanin antibodies and disodium cromolyn. BHK 21/C13 cell immunophenotype was assessed., Results: Fibrosarcoma cells were positive for vimentin, CD117, smooth muscle actin, vascular endothelial growth factor epidermal growth factor receptor, homebox prospero gene 1 and negative for platelet-derived growth factor B, neuron-specific enolase, S100, CD34, Ewing sarcoma and podoplanin. CAM-grown fibrosarcomas were highly sensitive to disodium cromolyn and anti-podoplanin antibodies., Conclusion: Immunophenotyping BHK 21/C13 cells and their response to drugs represent the first step in revealing cell line utility and a reliable tool for experimental cancer research., (Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Aim: To evaluate the impact of physical training on central hemodynamic parameters and elasticity of large arteries in hypertensive patients., Patients and Methods: A total of 129 hypertensive patients were divided into two groups: group A followed lifestyle changes and physical training; and group B acted as a control group; seven parameters were recorded: Pulse wave velocity (PWVao), systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure (PP), central aortic systolic blood pressure (SBPao), aortic diastolic blood pressure (DBPao), and central aortic pulse pressure (PPao)., Results: The difference between values at 4 months and baseline (Δ) were as follows: ΔPWVao was -1.02 m/s (p<0.001) versus 0.17 m/s (p=0.035), ΔSBPao was -9.6 mmHg (p=0.009) versus 1.6 mmHg (p=0.064), and ΔPPao was -6.8 mmHg (p<0.001) versus 3.2 mmHg, (p=0.029) in group A versus B, respectively., Conclusion: Exercise training improves SBP, PP, SBPao, PPao and may delay arterial ageing., (Copyright © 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Periodontal lesions are associated with activation of pathological angiogenesis and a high number of newly-formed blood vessels. Most angiogenic growth factors have been studied in the crevicular fluid or serum, but tissue correlations with vascular density or endothelial proliferation, are very rare, even inexistent. We assessed the VEGF/VEGFR2 axis expression in a multimodal fashion, in both epithelial and stromal compartments, with emphasis to endothelial proliferation and severity of periodontal lesions. Compared to normal gingiva, negative for VEGF/VEGFR2, periodontal lesions had a progressive increase for these markers from low to severe periodontal lesions. The transition from low to moderate periodontal lesions represents the milestone in disease progression and implies an active angiogenesis based on the highest angiogenic parameter variability observed for these lesions. Epithelial vascularization was firstly observed in moderate periodontal lesions and persists during severe periodontal disease. All the parameters used to quantify angiogenesis in periodontal lesions, were significantly increased in severe periodontal lesions dependent on VEGF expression in both the epithelial and stromal compartment. Our results support the use of anti-VEGF/VEGFR2-targeted therapy as adjuvant treatment for severe periodontal lesions., (Copyright © 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

The presence and distribution of lymphatic vessels and mast cells in the gingiva under normal and pathological conditions have been reported by several studies, but the relationship between them during inflammatory lymphangiogenesis is virtually unknown. The aim of the present study was to investigate the lymphatic microvessel density (LMVD) and mast cell density (MCD) in the gingiva of patients with periodontal disease compared to normal-like gingiva. Gingival punch biopsies from 51 patients with periodontal disease were investigated. MCs and LVs were detected by double-immunohistochemistry, using primary antibodies against mast cell tryptase and D2-40. The inflammatory infiltrate was evaluated on a scale from 0 (absent) to +3 (severe inflammation). MCs and LVs were counted in the same microscopic field for each case at ×200 magnification. We found a significant increase in the number of both MCs and LVs in cases with mild and moderate inflammatory changes, followed by a slight decrease in cases with severe inflammation. We have shown a particular association between MCs and LVs that may support the contribution of MCs to the development of the lymphatic vasculature in inflammatory conditions. MCD correlated with LMVD in all cases with mild and moderate inflammatory changes, but not in cases with severe inflammation. No correlation was found between MCD/LMVD and the density of the inflammatory infiltrate. Our results suggest the potential involvement of MCs in the induction and maintenance of lymphangiogenesis in the gingiva of patients with periodontal disease in the early steps of evolution., (Copyright © 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Aim: To investigate a possible correlation between the histological and morphometric properties of herniated intervertebral disc, clinical and magnetic resonance imaging (MRI) characteristics of patients with lumbar disc degeneration (LDD)., Materials and Methods: Thirty six patients with LDD were clinically evaluated using Japanese Orthopaedic Association Score (JOAS), visual analogue scale (VAS) for pain in the lower back or in the pelvic limb; MRI-based classification according to Pfirrmann and Modic criteria. All patients underwent decompressive surgery and herniated intervertebral disc samples were histologically and morphometrically analyzed. Data obtained were statistically analyzed for bivariate and partial correlations., Results: The mean area size of chondron clusters correlated with age, JOAS (r=-0.385, p=0.032, tau=-0.279, rho=-0.380), Pfirrmann (r=0.505, p=0.002, tau=0.289, rho=0.365) and Modic (r=0.500, p=0.002, tau=0.331, rho=0.419) grading. There was a strong correlation between maximum area size of chondrons and JOAS (r=-0.427, p=0.009, tau=-0.299, rho=-0.430), Pfirrmann changes (r=0.432, p=0.008, tau=0.309, rho=0.388) and Modic endplate changes (r=0.444, p=0.007, tau=0.343, rho=0.434). JOAS correlated with both MRI classifications used for LDD., Conclusion: The intervertebral disc cells tend to aggregate in clusters and the size of the chondrons from LDD correlated with JOAS, Pfirrmann and Modic. JOAS correlates with the imagistic evaluation systems Pfirrmann and Modic., (Copyright © 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Conchal cartilage or cartilage/ temporal fascia composite grafting (DC-F) used for rhinoplasty is applied by plastic surgeons for reconstructive purposes. Previous studies on experimental models such as mice or rabbits have elucidated on the late events following grafting, with tissue specimens being harvested two months after implantation. Early microscopic and molecular events following DC-F grafting are completely unknown. We designed a chick embryo chorioallantoic membrane model for human grafts study, regarding the dynamic observation of graft survival and its mutual interrelation with the chick embryo chorioallantoic membrane microenvironment. The DC-F graft preserved its cartilage component in a normal state compared to cartilage graft-only because of protective factors provided by temporal fascia. Its strong adherence to the cartilage, lack of angiogenic factors and high content of collagen IV-derived fragments with anti-angiogenic effects make the temporal fascia a good protective tissue to prevent implanted cartilage degeneration. The cartilage graft produced high inflammation, stromal fibrosis and activated angiogenic cascade through VEGF-mediated pathways followed by cartilage degeneration. Also, high content of podoplanin from conchal cartilage chondrocytes exerted a major role in inflammation accompanying cartilage graft. The presently employed experimental model allowed us to characterize the early histological and molecular events triggered by temporal fascia, cartilage or composite graft DC-F implanted on chick embryo chorioallantoic membrane. Our microscopic and molecular observations may help explain some post-surgical complications generated after using cartilage alone as biomaterial for nasal augmentation, supporting the use of DC-F composite graft, with the aim to reduce unwanted post-surgical events., (Copyright © 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Despite increasing knowledge on the cellular and molecular mechanisms involved in pulmonary fibrosis, its therapeutic options are still limited. The study of lymphangiogenesis has contributed to a better understanding of tumor growth and metastasis, with a major impact upon changes in therapeutic strategies and this was followed by the research of lymphatic vessels in other pathological conditions. Some data support the possible role of lymphangiogenesis in the pathogenesis of lung fibrosis. However, at the time of diagnosis for each patient with a fibrotic interstitial lung disease, it is necessary to predict the prognosis and to choose for individual targeted-therapy. Our aim was the characterization of lymphangiogenesis as a useful tool to stratify patients with lung fibrosis. We evaluated the presence, morphology and density of D2-40-positive lymphatic vessels and co-localization of D2-40/Ki67 in pulmonary fibrosis with different degrees of severity and without a specific etiology. Lymphatic vessel density did not correlate with severity grade and ranged between 4.66 to 38.33 vessels/×40 field, with the highest value in degree III of fibrosis. An intense proliferative activity of lymphatic endothelial cells was found in 24% (6 out of 25) of cases. The morphology of lymphatics and the presence of splitting combined with the proliferative activity of endothelial cell pillars suggested two different mechanisms in the formation new lymphatic vessels. Our results support the hypothesis that the activity and ongoing evolution of fibrosis can be predicted through the characterization of lymphangiogenesis but its presence or absence cannot predict the severity of fibrosis.

Podoplanin plays a crucial role for normal and pathological tissue development. Known as a lymphatic endothelial marker, podoplanin has been found to be overexpressed in tumor cells of various cancers with a certified involvement in tumor progression, invasion and metastasis. Oral cancer includes a heterogeneous group of malignancies with unpredictable behaviour and sometimes poor prognosis. Based on these facts, development of new molecular markers with a more reliable impact on therapy and prognosis is required. The present study was designed to characterize podoplanin expression in tumor cells of lip, oral cavity, tongue and pharynx squamous cell carcinomas, together with lymphatic vessels distribution, morphology, density and their impact on tumor progression. Evaluation of podoplanin by D2-40 immunohistochemistry assessement on 56 cases of oral cancers, revealed two different expression patterns in tumor cells depending on their location. Peri-tumor and intra-tumor lymphatic vessels density, morphology and distribution were correlated with lymph node status but not with tumor stage. The highest number of lymphatic vessels was observed in grade 3 squamous cell carcinomas. Dual expression of podoplanin in tumor cells and lymphatics with particular patterns correlated with histopathology and lymph node status in oral cancer, representing the molecular basis for testing podoplanin as a potential target for anti D2-40 antibody based therapy.

Aim: Intranodal palisaded myofibroblastoma (IPM) is a rare benign mesenchymal tumor restricted to the lymph nodes. Here, we report the case of a 44-year-old male patient with an IPM confined to the left laterocervical area., Case Report: After an accurate microscopic evaluation of morphological and histochemical stains, immunohistochemistry was performed for vimentin, smooth muscle actin vascular markers, S100 protein, D2-40, Ki67, lymphoid and melanoma markers, keratin and desmin on sections obtained from a paraffin-embedded surgical biopsy., Results: Spindle cell proliferation was positive for vimentin, smooth muscle cell actin and D2-40 and negative for the other markers. Low proliferative index, assessed by Ki67, was found. Based on morphological and immunohistochemical findings we diagnosed this case as intranodal palisaded myofibroblastoma and we highlighted a D2-40 expression in the tumor spindle cells. The presence of mast cells and their particular distribution inside the tumor are also, together with D2-40 expression, original findings of this study. No therapy was recommended after surgical and histopathological evaluation. The evolution of the patient was favorable with no other relapse following surgical removal of the lymphadenopathy. He has a normal life and no other changes of clinical and biological parameters were registered., Conclusion: To the best of our knowledge, this is the first report regarding a D2-40-positive reaction in the spindle cells of intranodal palisaded myofibroblastoma. Thus, D2-40 could be added to the panel of antibodies used for immunohistochemical diagnosis of such types of tumors.

Recently approved as treatment for astrocytoma, kidney and pancreatic cancer, everolimus acts on tumor cells by inhibiting tumor cell growth and proliferation, as well as by inhibition of angiogenic activity by both direct effects on vascular cell proliferation and indirect effects on growth factor production. The effects of everolimus on early stages of normal vasculogenesis, angiogenesis and lymphangiogenesis are not yet available. We found increased development of intravascular pillars by using area vasculosa of the chick chorioallantoic membrane treated with everolimus. An active lymphangiogenic response was highlighted by the expression of Prospero homeobox protein 1 (Prox1) and podoplanin, together with vascular endothelial growth factor receptor C (Vegf-C) and vascular endothelial growth factor receptor 3 (Vegfr-3) expression on day 4 in the treated group. These findings suggest a potential role of everolimus in the activation of lymphangiogenesis.

Background: Endocrine gland-related vascular endothelial growth factor (EG-VEGF), is an angiogenic factor specifically targeting endothelial cells derived from endocrine tissues. The inhibition of the EG-VEGF/prokineticin receptor pathway could represent a selective antiangiogenic and anticancer strategy., Aim: to evaluate the impact of an antibody to EG-VEGF on the rapidly growing capillary plexus of the chick embryo chorioallantoic membrane (CAM)., Materials and Methods: The in ovo CAM assay was performed for the humanized EG-VEGF antibody., Results: Hemorrhagic damage was induced in the capillaries, which led to early death of the embryos. Upon morphological staining, there was evidence of vascular disruption and extravasation of red blood cells in the chorion. Signs of vacuolization of the covering epithelium were also observed., Conclusion: Blocking endogenous EG-VEGF might represent a valuable approach of impairing or inhibiting angiogenesis in steroidogenic-derived embryonic tissues.