المؤلفون: Rod T. Mitchell, Kristine J. Hare, Mette Schou Hammerum, Cecilie Melau, Lene Lundvall, Lea Langhoff Thuesen, Anders Juul, Anne Jørgensen, Hanne Frederiksen, John E. Nielsen, Signe Perlman

المصدر: Melau, C, Nielsen, J E, Perlman, S, Lundvall, L, Langhoff Thuesen, L, Juul Hare, K, Schou Hammerum, M, Frederiksen, H, Mitchell, R T, Juul, A & Jørgensen, A 2021, ' Establishment of a Novel Human Fetal Adrenal Culture Model that Supports de Novo and Manipulated Steroidogenesis ', Journal of Clinical Endocrinology and Metabolism, vol. 106, no. 3, pp. 843-857 . https://doi.org/10.1210/clinem/dgaa852
Melau, C, Nielsen, J E, Perlman, S, Lundvall, L, Thuesen, L L, Hare, K J, Hammerum, M S, Frederiksen, H, Mitchell, R T, Juul, A & Jørgensen, A 2020, ' Establishment of a novel human fetal adrenal culture model that supports de novo and manipulated steroidogenesis ', Journal of Clinical Endocrinology & Metabolism . https://doi.org/10.1210/clinem/dgaa852

مصطلحات موضوعية: 0301 basic medicine, medicine.medical_specialty, steroidogenesis, Adrenal disorder, Cell Survival, Endocrinology, Diabetes and Metabolism, Primary Cell Culture, Clinical Biochemistry, ketoconazole, Drug Evaluation, Preclinical, 030209 endocrinology & metabolism, Adrenocorticotropic hormone, endocrine activity, Models, Biological, Biochemistry, 03 medical and health sciences, Fetus, 0302 clinical medicine, Endocrinology, ex vivo culture, Adrenocorticotropic Hormone, Pregnancy, Internal medicine, Adrenal Glands, medicine, Humans, Endocrine system, Congenital adrenal hyperplasia, androgen biosynthesis, Glucocorticoids, Adrenal Hyperplasia, Congenital, business.industry, Biochemistry (medical), medicine.disease, Culture Media, Androgen secretion, ACTH, Ketoconazole, 030104 developmental biology, Androgens, Female, Steroids, business, Metabolic Networks and Pathways, Ex vivo, Hormone

الوصف: Context Disorders affecting adrenal steroidogenesis promote an imbalance in the normally tightly controlled secretion of mineralocorticoids, glucocorticoids, and androgens. This may lead to differences/disorders of sex development in the fetus, as seen in virilized girls with congenital adrenal hyperplasia (CAH). Despite the important endocrine function of human fetal adrenals, neither normal nor dysregulated adrenal steroidogenesis is understood in detail. Objective Due to significant differences in adrenal steroidogenesis between human and model species (except higher primates), we aimed to establish a human fetal adrenal model that enables examination of both de novo and manipulated adrenal steroidogenesis. Design and Setting Human adrenal tissue from 54 1st trimester fetuses were cultured ex vivo as intact tissue fragments for 7 or 14 days. Main Outcome Measures Model validation included examination of postculture tissue morphology, viability, apoptosis, and quantification of steroid hormones secreted to the culture media measured by liquid chromatography-tandem mass spectrometry. Results The culture approach maintained cell viability, preserved cell populations of all fetal adrenal zones, and recapitulated de novo adrenal steroidogenesis based on continued secretion of steroidogenic intermediates, glucocorticoids, and androgens. Adrenocorticotropic hormone and ketoconazole treatment of ex vivo cultured human fetal adrenal tissue resulted in the stimulation of steroidogenesis and inhibition of androgen secretion, respectively, demonstrating a treatment-specific response. Conclusions Together, these data indicate that ex vivo culture of human fetal adrenal tissue constitutes a novel approach to investigate local effects of pharmaceutical exposures or emerging therapeutic options targeting imbalanced steroidogenesis in adrenal disorders, including CAH.

وصف الملف: application/pdf

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7973a5c971517825f03b64cfda648bb0
https://curis.ku.dk/portal/da/publications/establishment-of-a-novel-human-fetal-adrenal-culture-model-that-supports-de-novo-and-manipulated-steroidogenesis(8658a448-6b64-4c00-a127-6518152fb667).html

المؤلفون: Richard J. Auchus, Juilee Rege, William E. Rainey, Tobias Else, Adina F. Turcu

المصدر: J Steroid Biochem Mol Biol

مصطلحات موضوعية: 0301 basic medicine, medicine.medical_specialty, Adrenal disorder, medicine.drug_class, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, Clinical Biochemistry, Adrenal Gland Diseases, Adrenocorticotropic hormone, Biochemistry, Article, Steroid, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Endocrinology, Internal medicine, Adrenal Glands, medicine, Humans, Molecular Biology, business.industry, Cholesterol, Cell Biology, Androgen, Angiotensin II, Biosynthetic Pathways, 030104 developmental biology, chemistry, 030220 oncology & carcinogenesis, Molecular Medicine, Biomarker (medicine), Steroids, business, Biomarkers, Hormone

الوصف: Adrenal steroidogenesis is a robust process, involving a series of enzymatic reactions that facilitate conversion of cholesterol into biologically active steroid hormones under the stimulation of angiotensin II, adrenocorticotropic hormone and other regulators. The biosynthesis of mineralocorticoids, glucocorticoids, and adrenal-derived androgens occur in separate adrenocortical zones as a result of the segregated expression of steroidogenic enzymes and cofactors. This mini review provides the principles of adrenal steroidogenesis, including the classic and under-appreciated 11-oxygenated androgen pathways. Several adrenal diseases result from dysregulated adrenal steroid synthesis. Herein, we review growing evidence that adrenal diseases exhibit characteristic modifications from normal adrenal steroid pathways that provide opportunities for the discovery of biomarker steroids that would improve diagnosis and monitoring of adrenal disorders.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0b6382f50c20561a305f4bcc0d090c7c
https://doi.org/10.1016/j.jsbmb.2019.01.018

المؤلفون: Therapeutics, Jon M. Nakamoto, Rose Marino, Sejal Shah, Bradley S. Miller, Kyriakie Sarafoglou, Manmohan K. Kamboj, Molly O. Regelmann, Takara L. Stanley

المصدر: The Journal of Clinical Endocrinology & Metabolism. 103:4324-4331

مصطلحات موضوعية: Male, 0301 basic medicine, medicine.medical_specialty, Pediatrics, Hydrocortisone, Adrenal disorder, Endocrinology, Diabetes and Metabolism, Clinical Biochemistry, Context (language use), Biochemistry, Diagnostic Techniques, Endocrine, 03 medical and health sciences, Neonatal Screening, 0302 clinical medicine, Endocrinology, Adrenocorticotropic Hormone, Reference Values, Internal medicine, medicine, Adrenal insufficiency, Humans, Adrenoleukodystrophy, Aldosterone, Societies, Medical, Newborn screening, business.industry, Biochemistry (medical), Infant, Newborn, Adrenal crisis, medicine.disease, 030104 developmental biology, Systematic review, North America, medicine.symptom, Age of onset, business, 030217 neurology & neurosurgery, Adrenal Insufficiency

الوصف: Context Adrenoleukodystrophy (ALD) is a peroxisomal disorder associated with neurologic decompensation and adrenal insufficiency. Newborn screening for ALD has recently been implemented in five states with plans to expand to all 50 states in the United States. Adrenal insufficiency ultimately develops in most males with ALD, but the earliest age of onset is not well established. Objective These clinical recommendations are intended to address screening for adrenal insufficiency in boys identified to have ALD by newborn screen. Participants Seven members of the Pediatric Endocrine Society Drug and Therapeutics/Rare Diseases Committee, with clinical experience treating children with ALD and adrenal insufficiency, and a pediatric endocrinologist and laboratory director were selected to be on the working committee. Consensus Process The authors comprised the working group and performed systematic reviews of the published literature regarding adrenal insufficiency and ALD. The recommendations were reviewed and approved by the larger Pediatric Endocrine Society Drug and Therapeutics/Rare Diseases Committee and then by the Pediatric Endocrine Society Board of Directors. Conclusions There is limited literature evidence regarding monitoring of evolving adrenal insufficiency in male infants and children with ALD. The recommendations suggest initiating assessment of adrenal function at diagnosis with ALD and regular monitoring to identify boys with adrenal insufficiency in a timely manner and prevent life-threatening adrenal crisis. These recommendations are intended to serve as an initial guide, with the understanding that additional experience will inform future guidelines.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e477b1e1c7d3cbb2c0d99d70f311acf0
https://doi.org/10.1210/jc.2018-00920

المؤلفون: Raman Deep Singh, Ravinder J. Singh, Irina Bancos, William F. Young, Aditya V Avula, Cristian Bancos, Jolaine M. Hines, Stefan K.G. Grebe

المصدر: Clinical Chemistry. 63:1824-1835

مصطلحات موضوعية: 0301 basic medicine, medicine.medical_specialty, Adrenal disorder, medicine.drug_class, Chemistry, Metabolite, Biochemistry (medical), Clinical Biochemistry, 030209 endocrinology & metabolism, Urine, medicine.disease, Androgen, Excretion, 03 medical and health sciences, Cushing syndrome, chemistry.chemical_compound, 030104 developmental biology, 0302 clinical medicine, Endocrinology, Internal medicine, medicine, Congenital adrenal hyperplasia, Glucocorticoid, medicine.drug

الوصف: BACKGROUND Steroid profiling is a promising diagnostic tool with adrenal tumors, Cushing syndrome (CS), and disorders of steroidogenesis. Our objective was to develop a multiple-steroid assay using liquid-chromatography, high-resolution, accurate-mass mass spectrometry (HRAM LC-MS) and to validate the assay in patients with various adrenal disorders. METHODS We collected 24-h urine samples from 114 controls and 71 patients with adrenal diseases. An HRAM LC-MS method was validated for quantitative analysis of 26 steroid metabolites in hydrolyzed urine samples. Differences in steroid excretion between patients were analyzed based on Z-score deviation from control reference intervals. RESULTS Limits of quantification were 20 ng/mL. Dilution linearity ranged from 80% to 120% with means of 93% to 110% for all but 2 analytes. Intraassay and interassay imprecision ranged from 3% to 18% for all but 1 analyte. Control women had lower excretion of androgen and glucocorticoid precursors/metabolites than men (P < 0.001), but no difference in mineralocorticoids was seen (P = 0.06). Androgens decreased with age in both sexes (P < 0.001). Compared with patients with adrenocortical adenoma (ACA), patients with adrenocortical carcinoma (ACC) had 11 steroids with increased Z scores, especially tetrahydro-11-deoxycortisol (14 vs 0.5, P < 0.001), pregnanetriol (7.5 vs −0.4, P = 0.001), and 5-pregnenetriol (5.4 vs −0.4, P = 0.01). Steroid profiling also demonstrated metabolite abnormalities consistent with enzymatic defects in congenital adrenal hyperplasia and differences in pituitary vs adrenal CS. CONCLUSIONS Our HRAM LC-MS assay successfully quantifies 26 steroids in urine. The statistically significant differences in steroid production of ACC vs ACA, adrenal vs pituitary CS, and in congenital adrenal hyperplasia should allow for improved diagnosis of patients with these diseases.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::c80602f28fb6bbc535591f05b2eae1bd
https://doi.org/10.1373/clinchem.2017.271106

المؤلفون: Aya T Nanba, Juilee Rege, Adina F. Turcu, Jianwei Ren, Hwei Ming Peng, William E. Rainey, Richard J. Auchus

المصدر: The Journal of Clinical Endocrinology & Metabolism. 103:320-327

مصطلحات موضوعية: Adult, Male, 0301 basic medicine, endocrine system, medicine.medical_specialty, Adrenal disorder, Endocrinology, Diabetes and Metabolism, Clinical Biochemistry, 030209 endocrinology & metabolism, Stimulation, Adrenocorticotropic hormone, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Endocrinology, Dehydroepiandrosterone sulfate, Adrenocorticotropic Hormone, Zona fasciculata, Cosyntropin, Internal medicine, Adrenal Glands, Hyperaldosteronism, medicine, Humans, Clinical Research Articles, Cells, Cultured, Aged, Dehydroepiandrosterone Sulfate, Biochemistry (medical), Middle Aged, Prognosis, 030104 developmental biology, medicine.anatomical_structure, chemistry, Case-Control Studies, Pregnenolone, Female, Pregnenolone sulfate, hormones, hormone substitutes, and hormone antagonists, Zona reticularis, Follow-Up Studies

الوصف: Background Dehydroepiandrosterone sulfate (DHEAS) is the most abundant steroid in human circulation, and adrenocorticotropic hormone (ACTH) is considered the major regulator of its synthesis. Pregnenolone sulfate (PregS) and 5-androstenediol-3-sulfate (AdiolS) have recently emerged as biomarkers of adrenal disorders. Objective To define the relative human adrenal production of Δ5-steroid sulfates under basal and cosyntropin-stimulated conditions. Methods Liquid chromatography-tandem mass spectrometry was used to quantify three unconjugated and four sulfated Δ5-steroids in (1) paired adrenal vein (AV) and mixed venous serum samples (21 patients) and (2) cultured human adrenal cells both before and after cosyntropin stimulation, (3) microdissected zona fasciculata (ZF) and zona reticularis (ZR) from five human adrenal glands, and (4) a reconstituted in vitro human 17α-hydroxylase/17,20-lyase/(P450 17A1) system. Results Of the steroid sulfates, PregS had the greatest increase after cosyntropin stimulation in the AV (32-fold), whereas DHEAS responded modestly (1.8-fold). PregS attained concentrations comparable to those of DHEAS in the AV after cosyntropin stimulation (AV DHEAS/PregS, 24 and 1.3 before and after cosyntropin, respectively). In cultured adrenal cells, PregS demonstrated the sharpest response to cosyntropin, whereas DHEAS responded only modestly (21-fold vs 1.8-fold higher compared with unstimulated cells at 3 hours, respectively). Steroid analyses in isolated ZF and ZR showed similar amounts of PregS and 17α-hydroxypregnenolone in both zones, whereas DHEAS and AdiolS were higher in ZR (P < 0.05). Conclusion Our studies demonstrated that unlike DHEAS, PregS displayed a prominent acute response to cosyntropin. PregS could be used to interrogate the acute adrenal response to ACTH stimulation and as a biomarker in various adrenal disorders.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5f6762513d02c8563e636dbc3c821bd5
https://doi.org/10.1210/jc.2017-01525

المؤلفون: Zev Rosenwaks, David E. Reichman

المصدر: The Journal of Steroid Biochemistry and Molecular Biology. 165:131-136

مصطلحات موضوعية: Male, Ovulation, Human fertility, Infertility, medicine.medical_specialty, Adrenal disorder, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, media_common.quotation_subject, DNA Mutational Analysis, Clinical Biochemistry, Adrenal Gland Diseases, 030209 endocrinology & metabolism, Fertility, Biology, Bioinformatics, Biochemistry, Steroid, Mice, 03 medical and health sciences, Human reproduction, 0302 clinical medicine, Endocrinology, Internal medicine, medicine, Animals, Humans, Gonads, Molecular Biology, media_common, 030219 obstetrics & reproductive medicine, Adrenal Hyperplasia, Congenital, Reproduction, Cell Biology, medicine.disease, Hormones, Steroid hormone, Mutation, Androgens, Steroid 11-beta-Hydroxylase, Molecular Medicine, Female, Steroids

الوصف: Human fertility requires an exquisitely complex orchestration of steroid hormone action to affect the necessary elements of reproduction, including folliculogenesis, endometrial advancement, ovulation, and implantation. Individuals affected by genetic steroid disorders often face substantial challenges to these crucial elements of fertility, in addition to the broader health implications of their diseases. In the following article, we review the impact of genetic steroid disorders on human reproduction, as well as the treatments, where available, aimed at circumventing such hurdles. Adrenal disorders will first be described, followed by rare gonadal steroid disorders.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::117b100191e434a9cdd8b2a7055f9af2
https://doi.org/10.1016/j.jsbmb.2016.04.014

المؤلفون: Holger S. Willenberg

المصدر: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme. 49(3)

مصطلحات موضوعية: medicine.medical_specialty, Hypertension, Renal, Adrenal disorder, Endocrinology, Diabetes and Metabolism, Clinical Biochemistry, MEDLINE, Secondary hypertension, 030209 endocrinology & metabolism, Disease, 030204 cardiovascular system & hematology, Sodium Chloride, Biochemistry, SALT RETENTION, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Primary aldosteronism, Internal medicine, Health care, Hyperaldosteronism, medicine, Humans, Medical treatment, business.industry, Biochemistry (medical), Genetic Diseases, Inborn, General Medicine, medicine.disease, business

الوصف: The last years have seen substantial progress in primary aldosteronism (PA), which is the most common cause of secondary hypertension. Many programs have been established around the world to meet the needs in healthcare and the management of patients with PA according to published guidelines and clinical protocols. Systematic analysis of emerging data and meticulous scientific work have informed us on the molecular basis of the disease and helped to characterize hereditary forms of PA. Techniques have been developed to better diagnose PA and to establish genotype-phenotype relationships and their impact on hypertension. Studies have been undertaken to stratify patients for risk factors and to ensure quality of best medical treatment. This review focuses on some clinically relevant problems in characterizing autonomous aldosterone secretion and discusses testing and management strategies. Besides, this review puts the emphasis on some colorful studies not to pale soon beside an ever evolving painting background.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fe3789f8f5947e1ce6d3064e597a1652
https://pubmed.ncbi.nlm.nih.gov/28351083

المؤلفون: Jha, Deepti Goswami, Ayesha

المصدر: Journal of Clinical and Diagnostic Research, Vol 10, Iss 10, Pp QC10-QC12 (2016)

مصطلحات موضوعية: 0301 basic medicine, endocrine system, medicine.medical_specialty, endocrine system diseases, Adrenal disorder, Clinical Biochemistry, lcsh:Medicine, Dehydroepiandrosterone, Blood sugar, Obstetrics and Gynaecology Section, amenorrhea, 03 medical and health sciences, 0302 clinical medicine, Internal medicine, medicine, Autoimmune disease, 030219 obstetrics & reproductive medicine, business.industry, autoimmunity, lcsh:R, Thyroid, General Medicine, thyroid autoantibodies, medicine.disease, Anti-thyroid autoantibodies, Premature ovarian failure, 030104 developmental biology, medicine.anatomical_structure, Endocrinology, Amenorrhea, medicine.symptom, business

الوصف: Introduction Premature Ovarian Failure (POF) is the cessation of ovarian function before the age of 40 years. POF is reported to be associated with autoimmune diseases in 20-30% of cases. Aim Patients presenting with idiopathic POF were screened for the presence of autoimmune disorders. Materials and methods Twenty patients with idiopathic POF were included in the study. Baseline investigation in all subjects included fasting serum FSH, LH, E2, progesterone, free T3, free T4, Thyroid-Stimulating Hormone (TSH) and Anti-Thyroperoxidase (anti-TPO) antibodies, testosterone and Dehydroepiandrosterone (DHEAS) levels. Fasting and post-glucose (2 hours after 75g of oral glucose) serum calcium and phosphate were estimated using appropriate assays in biochemistry laboratory. Results Seven patients (35%), who presented with secondary amenorrhea, had thyroid disorders and were already on thyroxine replacement therapy. One patient also had vitiligo. There was no history of adrenal disorder. Anti-TPO levels were elevated in two (10%) patients of secondary amenorrhea group. The levels of serum testosterone were low in three patients. Serum DHEAS levels were low in 13 patients. Blood sugar levels (fasting and 2 hour post 75g glucose load) and fasting insulin levels were normal. Serum calcium and phosphate levels were normal in all the patients. Conclusion Thyroid autoimmunity is the most common autoimmune disease associated with POF. The finding of low DHEAS in a large percentage of patients (65%), suggests possibility of adrenal dysfunction. This requires further testing for adrenal reserve and adrenal autoantibodies.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::67febcf23c648ce0b8e1b91910096b10
https://pubmed.ncbi.nlm.nih.gov/27891401

المؤلفون: Marc Lombès, Claire Bouvattier, Pascal Boileau, Laetitia Martinerie, Simon Travers, Eric Pussard

المساهمون: Signalisation Hormonale, Physiopathologie Endocrinienne et Métabolique, AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Sud - Paris 11 (UP11), AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Risques cliniques et sécurité en santé des femmes et en santé périnatale (RISCQ), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut Biomédical de Bicêtre (US 32), Institut National de la Santé et de la Recherche Médicale (INSERM)-AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Domaine d’Intérêt Majeur Logiciels et Systèmes Complexes Society for Endocrinology Institut National de la Santé et de la Recherche Médicale, S.T was recipient of a fellowship from the French Endocrine Society (SFE). This work has been supported by fundings from Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Sud, Assistance Publique de Paris. We also acknowledge the funding from a Cardiovasculaire-Obésité-Rein-Diabète-Domaine d’Intérêt Majeur (CORDDIM, Région Ile-de-France) grant that partly supported the acquisition of the Waters Xevo TQS triple-quadrupole mass spectrometer. We gratefully thank Prof. William Reinus for helpful comments and language editing of the manuscript. The authors would like to thank Valerie Steiner and Cécile Dupuy for their advice in the first steps of method development.

المصدر: Journal of Steroid Biochemistry and Molecular Biology
Journal of Steroid Biochemistry and Molecular Biology, Elsevier, 2017, 165, pp.202-211. ⟨10.1016/j.jsbmb.2016.06.005⟩

مصطلحات موضوعية: 0301 basic medicine, Adrenal disorder, [SDV]Life Sciences [q-bio], Endocrinology, Diabetes and Metabolism, Clinical Biochemistry, Steroid profiling, 01 natural sciences, Biochemistry, chemistry.chemical_compound, Endocrinology, Liquid chromatography–mass spectrometry, Corticosterone, Limit of Detection, Tandem Mass Spectrometry, Child, Aldosterone, Chromatography, High Pressure Liquid, Immunoassay, 3. Good health, Molecular Medicine, Regression Analysis, Steroids, medicine.drug, medicine.medical_specialty, 03 medical and health sciences, Dehydroepiandrosterone sulfate, LC–MS/MS, Internal medicine, Mineralocorticoids, medicine, Humans, Congenital adrenal hyperplasia, Molecular Biology, Glucocorticoids, Chromatography, Adrenal Hyperplasia, Congenital, 010401 analytical chemistry, Selected reaction monitoring, Reproducibility of Results, Cell Biology, medicine.disease, 0104 chemical sciences, 030104 developmental biology, chemistry, Mutation, Cortisone, Chromatography, Liquid

الوصف: International audience; Serum steroid assays are major tools in the clinical evaluation of adrenal disorders. The main adrenal steroids are routinely measured with immunoassays. However, chromatographic methods are known to offer better specificity. We report a liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay for simultaneous quantification of 15 adrenal steroids targeting the mineralo- and gluco-corticosteroid pathways. Serum steroids combined with deuterated internal standards were extracted using successive protein precipitation and solid phase extraction steps. Cortisol, cortisone, 11-deoxycortisol, 17-hydroxyprogesterone, 21-deoxycortisol, progesterone, 11-deoxycorticosterone, corticosterone, 11-dehydrocorticosterone, 18-hydroxycorticosterone, 18-hydroxy-11-deoxycorticosterone, aldosterone, dehydroepiandrosterone sulfate, testosterone and androstenedione were resolved in fourteen minutes using a BEH C18 column coupled to a methanol-ammonium formate gradient. Detection was performed using multiple reaction monitoring quantitation. Routinely determined steroid levels by immunoassays were compared to those measured by LC–MS/MS. This method was applied to assess steroid profiles in congenital adrenal hyperplasia (CAH) patients with 21-hydroxylase deficiency. Low quantification limits depending on each steroid (ranging from 0.015 ng/mL for aldosterone to 20 ng/mL for DHEAS) are adapted to the clinical use. Recoveries of steroids range from 64% for 21-deoxycortisol to 101% for cortisol and are fully corrected by internal standards. A good linearity with R > 0.989 is obtained for each compound. The inter-day variation coefficients ranged from 4.7% for cortisol to 16.3% for 11-deoxycorticosterone. The immunoassay for cortisol (Immulite 2000, Siemens) showed acceptable agreement with LC–MS/MS (bias +7.2%). However, Bland-Altman plots revealed large negative bias for aldosterone (−33.4%, AldoCT, CisBio international), for 17-hydroxyprogesterone at concentrations below 2 ng/mL (−74.1%, OHP-CT MP Biomedical), for androstenedione (−80.3%, RIA D4, Beckman Coulter) and for 11-deoxycortisol (−125.3%, Diasource Immunoassays). Finally, the analysis of samples from 21-hydroxylase defective patients demonstrated the potential usefulness of multiplexed steroid profiling for the diagnosis and/or monitoring of different forms of congenital adrenal hyperplasia. This LC–MS/MS method provides highly sensitive and specific assessments of mineralo- and glucocorticoids pathways from a small volume sample and is therefore a promising potent tool for clinical and experimental endocrine studies.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8d0be30c0745955c63a2997621c51223
https://pubmed.ncbi.nlm.nih.gov/27339652

المؤلفون: Patricia Cogram, Bruno Ferraz-de-Souza, Yves Morel, Dianne Gerrelli, Franziska Martin, Felix Beuschlein, Angela Huebner, John C. Achermann, Lin Lin, Delphine Mallet, Rebecca E. Hudson-Davies

المصدر: The Journal of Clinical Endocrinology & Metabolism. 94:678-683

مصطلحات موضوعية: Male, Steroidogenic factor 1, medicine.medical_specialty, Adrenal disorder, Receptors, Retinoic Acid, Endocrinology, Diabetes and Metabolism, DNA Mutational Analysis, Clinical Biochemistry, Adrenal Gland Diseases, Biology, Steroidogenic Factor 1, Transfection, Biochemistry, Transactivation, Endocrinology, Proto-Oncogene Proteins, Internal medicine, Adrenal Glands, medicine, Humans, Pre-B-Cell Leukemia Transcription Factor 1, Transcription factor, Cells, Cultured, DAX-1 Orphan Nuclear Receptor, Adrenal hypoplasia, Biochemistry (medical), Gene Expression Regulation, Developmental, Adrenal Cortex Neoplasm, DNA-Binding Proteins, Gene Expression Regulation, Neoplastic, Repressor Proteins, Mutation, Trans-Activators, Original Article, Female, DAX1

الوصف: Context: Disorders of adrenal development result in significant morbidity and mortality. However, the molecular basis of human adrenal development, and many forms of disease, is still poorly understood.Objectives: We evaluated the role of two new candidate genes, CBP/p300-interacting transactivator, with Glu/Asp-rich C-terminal domain, 2 (CITED2), and pre-B-cell leukemia transcription factor 1 (PBX1), in human adrenal development and disease.Design: CITED2 and PBX1 expression in early human fetal adrenal development was assessed using RT-PCR and in situ hybridization. The regulation of CITED2 and PBX1 by steroidogenic factor-1 (SF-1) and dosage-sensitive sex reversal, adrenal hypoplasia congenital, critical region on the X chromosome, gene-1 (DAX1) was evaluated in NCI-H295R human adrenocortical tumor cells by studying promoter regulation. Finally, mutational analysis of CITED2 and PBX1 was performed in patients with primary adrenal disorders.Results: CITED2 and PBX1 are expressed in the human fetal adrenal gland during early development. Both genes are activated by SF-1 in a dose-dependent manner in NCI-H295R cells, and, surprisingly, PBX1 is synergistically activated by SF-1 and DAX1. Mutational analysis failed to reveal significant coding sequence changes in individuals with primary adrenal disorders.Conclusions: CITED2 and PBX1 are likely to be important mediators of adrenal development and function in humans, but mutations in these genes are not common causes of adrenal failure in patients in whom a molecular diagnosis remains unknown. The positive interaction between DAX1 and SF-1 in regulating PBX1 may be an important mechanism in this process.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::439bd1435dd3dea42c774cd6b27325ac
https://doi.org/10.1210/jc.2008-1064