Citations
All
Search in:AllTitleAbstractAuthor name
Publications
(716)
Patents
Grants
Pathways
Clinical trials
Journal: Clinical Chemistry and Laboratory Medicine
January/11/2020
Abstract
Background Classically, serum testosterone (T) and androstenedione (A4) have been the mainstay for the biochemical assessment of hyperandrogenism. However, recent evidence suggests 11β-hydroxyandrostenedione (11OHA4) and 11-ketotestosterone (11KT) may also be important. Here, we describe the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for quantitation of total serum T, A4, 17-hydroxyprogesterone (<em>17OHP</em>), 11OHA4 and 11KT. In addition, we applied the method to assess pre-analytical stability. Methods An isotopically labelled internal standard was added to samples prior to supported liquid extraction (SLE). Extracts were analysed using LC-MS/MS to detect T/A4/<em>17OHP</em>/11OHA4 and 11KT along with their corresponding internal standards. Samples (n = 7) were collected from healthy volunteers (n = 14) and left incubated at 20 °C for up to 72 h. Tubes were retrieved at select time points, centrifuged, separated and frozen prior to analysis. Results The total run time was 4 min. For all analytes, intra- and inter-assay imprecision did not exceed 7.9% and 5.3%, respectively; matrix effects were negligible and mean recoveries ranged from 95.3 to 111.6%. The limits of quantitation (LOQs) were 0.25 nmol/L for T, A4 and 11OHA4, 0.50 nmol/L for <em>17OHP</em>, and 0.24 nmol/L for 11KT. No significant change was observed in pre-centrifugation A4 or female T concentrations over 72 h. Significant increases (p < 0.01) in concentrations of 11KT, <em>17OHP</em>, 11OHA4 and male T were observed after 2, 8, 12 and 24 h, respectively. Conclusions We developed a robust LC-MS/MS assay for the quantitation of total serum T/A4/<em>17OHP</em>/11OHA4 and 11KT. Applying the method to determine pre-analytical stability suggests samples requiring 11KT need separating from the cells within 2 h.
Related with
Journal: Clinical Endocrinology
May/9/2016
Abstract
BACKGROUND
A physiological increase in androgen levels occurs during adolescence. Measuring androgen concentrations is the best method to distinguish normal evolution processes from hyperandrogenic disorders.
OBJECTIVE
The increase in circulating androgens during puberty is inversely associated with insulin sensitivity in normal weight girls.
OBJECTIVE
To assess circulating levels of ovarian androgens and anti-Müllerian hormone (AMH) at baseline and after GnRH analogue (GnRH-a) stimulation in normal pubertal girls across different Tanner stages. We also studied the association between this response and insulin sensitivity.
METHODS
Prospective study of healthy girls (6-12 years) from the local community (n = 63).
METHODS
Tanner I (n = 23) subjects were assessed cross-sectionally, and Tanner II girls (n = 40) were evaluated every 6 months until they reached Tanner V. Early morning dehydroepiandrosterone sulphate (DHEA-S), AMH, sex hormone-binding globulin (SHBG), androstenedione, glucose and insulin levels were measured. A GnRH-a test (500 μg/m(2) ; sc) and oral glucose intolerance test (OGTT) were performed. Differences throughout puberty were evaluated.
RESULTS
Basal and/or stimulated Testosterone DHEA-S and 17-hydroxyprogesterone (<em>17OHP</em>) were inversely associated with insulin sensitivity (WIBSI) from the beginning of puberty, whereas androstenedione was directly associated with gonadotrophins. AMH was inversely associated with basal and stimulated gonadotrophins and directly with insulin area under the curve (AUC) only in the early stages of puberty. <em>17OHP</em> and testosterone responsiveness increased significantly during puberty in all subjects, whereas testosterone levels changed less consistently. This pattern of ovarian-steroidogenic response was most evident during mid- and late puberty. Moreover, during late puberty only, basal <em>17OHP</em>, testosterone and DHEA-S were positively associated with gonadotrophins.
CONCLUSIONS
In normal nonobese girls born appropriate for gestational age, androgen synthesis was associated with insulin sensitivity in early puberty and with LH only in late puberty.
Journal: Hormone research
July/23/1997
Abstract
Monitoring therapy for congenital adrenal hyperplasia (CAH) due to 21-hydroxylase is difficult, although plasma determinations of 17 alpha-hydroxyprogesterone (<em>17OHP</em>), delta 4-androstenedione (delta 4A) and testosterone are helpful. We have studied the usefulness of monitoring plasma 3 alpha-androstanediol glucuronide (3 alpha-AG) in group of 24 CAH patients aged from birth to 18 years. For comparison, normal values for age and pubertal stage were determined in a control group of 115 girls and 118 boys. Mean plasma levels were higher during the first year of life, decreased to a nadir between 1 and 4 years, and increased steadily thereafter, there was also a significant increase with pubertal stage. In 24 pairs of blood samples obtained at the time of venopuncture and 2 h after, 3 alpha-AG levels did not change (p>> 0.05) demonstrating that 3 alpha-AG levels were not affected by stress. In the patients with CAH, positive correlations between plasma 3 alpha-AG and delta 4A (females, r = 0.73; males, r = 0.98), <em>17OHP</em> (females, r = 0.58; males, r = 0.84) and testosterone (females, r = 0.83; males, r = 0.97) were observed. Concordance between 3 alpha-AG and delta 4A was observed in 90% of all samples, and in 91% between 3 alpha-AG and testosterone. Our study demonstrates that 3 alpha-AG is a valid marker of control and its determination appears to be a reliable tool to monitor CAH.
Journal: Journal of Obstetrics and Gynaecology
October/11/2020
Abstract
<strong class="sub-title"> Objective: </strong> To determine whether <i>other</i> androgens [androstenedione (A4), 17-hydroxy progesterone (<em>17OHP</em>) and dehydroepiandrosterone (DHEA)] were elevated in women with classically defined idiopathic hirsutism (IH)/patient-important hirsutism (PIH).
Study design: Retrospective analysis.
Setting: Outpatient endocrine department of a tertiary care hospital.
<strong class="sub-title"> Patients: </strong> In total, 30 consecutive women with IH/PIH were included. IH/PIH was defined as presentation with hirsutism with normal menstrual cycles (25-35 days), normal total (< 45 ng/dL) and free T (fT) (< 0.6 ng/dL) and normal ovaries sonologically (transabdominal ultrasonogram ovarian volume < 10 cm<sup>3</sup>) without any other signs of virilization. Clinical and biochemical details were collected and analyzed. Androgens were measured by LC-MS/MS. A4 ≥ 2.5 ng/mL, DHEA ≥ 15 (age < 18) or ≥ 11.8 (age ≥ 18) ng/mL, DHEAS ≥ 2847 ng/mL or <em>17OHP</em> ≥ 2 ng/mL were considered high.
Results: With the mean age of 22 years and mean BMI of 25 kg/m2, 12/30 (40%) had IH and remaining PIH. DHEA alone was elevated in 60% and A4 alone in 33%. Overall, 23/30 (73%) had any one elevated androgen with normal total and free testosterone. There was no correlation with modified Ferriman-Gallwey score, and there was no significant difference in androgens between IH and PIH.
Conclusion: A high proportion of women with classically defined IH/PIH have elevated DHEA and/or A4. Though on pharmacotherapy basis, there would be no change in management, the role of hyperandrogenemia detected by sensitive assays on metabolic functions and cardiovascular risk has to be studied.
Keywords: Androstenedione; Dehydroepiandrosterone; Liquid chromatography–mass spectroscopy; Patient-important hirsutism.
Journal: Arquivos brasileiros de endocrinologia e metabologia
July/4/2006
Abstract
We evaluated linear growth of 27 children with congenital adrenal hyperplasia (CAH) treated with low doses of oral hydrocortisone. They were followed-up during 6.1 +/- 1.8 years with daily hydrocortisone doses of 10.84 +/- 2.0 mg/m2 and 0.1 mg fludrocortisone (24 of them). Twenty-three were female. Mean chronological age (CA) was 6.1 +/- 2.9 years and bone age (BA) 6.9 +/- 3.3 (r = 0.66) at the beginning of the study. Five children showed BA advancement>> 2 years relating to CA. It was calculated Height SD for CA (SD/H) and for BA (SD/BA) were calculated using NCHS as reference pattern. At the beginning of the study SD/H was -0.8 +/- 1.9 and corresponding SD/BA was -1.5 +/- 2.1; at the end SD/H was -0.17 +/- 1.5 and SD/BA was -1.34 +/- 1.2 (p = 0.02 and p = 0.51, respectively for the beginning and the end). BA changed 1.3 +/- 0.3 per year during this period. Children with advanced BA showed an improvement of SD/BA, from -4.55 +/- 0.9 at from the beginning, -4.55 +/- 0.9 to -2.48 +/- 0.4 at the end of follow-up, -2.48 +/- 0.4 (p = 0.003). The elevated plasma levels of 17-OH Progesterone (<em>17OHP</em>) and androstenedione showed further increase during follow-up. We conclude that children with CAH receiving low doses of hydrocortisone showed adequate growth during the follow-up, without excessive BA advancement, even though full suppression of plasma levels of <em>17OHP</em> and androgens wasere not achieved.
Journal: Revista Medica de Chile
June/20/2001
Abstract
BACKGROUND
The early diagnosis and therapy of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency can prevent adrenal crises and erroneous gender assignment in affected newborns. To achieve this goal neonatal mass-screening programs have been developed, measuring blood 17 alpha-hydroxyprogesterone (<em>17OHP</em>). In Chile there is no experience with this type of screening.
OBJECTIVE
To develop a method for measuring <em>17OHP</em> in filter paper blood specimens. To obtain reference ranges and determine neonatal <em>17OHP</em> threshold levels according to gestational age and birth weight. To analyze factors affecting the cost-efficiency ratio and suggest recommendations for the organization of a neonatal screening program for CAH in Chile.
METHODS
Nine hundred twenty two newborns were studied. <em>17OHP</em> was measured using double antibody radioimmunoassay in filter paper blood samples obtained 48 h after birth. Reference ranges were determined according to gestational age and birth weight and a cutoff point of 25 ng/ml was established.
RESULTS
Seventeen newborns had <em>17OHP</em> over the cutoff value. They were assessed by a pediatric endocrinologist and in none of them, CAH was confirmed. Therefore the false positive rate of the determination was 1.8%. Among these newborns with elevated <em>17OHP</em>, 66% had a birth weight below 1.5 kg and 5.8%, a birth weight between 1.5 and 2.5 kg. The cost per reported result was US $ 1. Timing of the recall was between the 3 and 10 days of life. No newborn missed the follow-up.
CONCLUSIONS
To increase the cost-efficiency ratio of an eventual neonatal screening program, newborns with birth weights below 1.5 kg should be excluded and cutoff points should be defined according to birth weight (Rev Méd Chile 2000; 128: 1113-18).
Journal: Journal of Pediatric Endocrinology and Metabolism
May/21/2003
Abstract
An increase in plasma <em>17OHP</em> found in infants requiring differential diagnosis between septic shock and adrenal failure led us to look for adrenal steroids pattern during infection.
METHODS
56 infants, 1-6 months old, were studied during infection of different degrees of severity. Plasma cortisol, <em>17OHP</em>, androstenedione, DHEA, DHEA-S and testosterone were measured.
RESULTS
24 patients showed an expected cortisol elevation. One child had a low cortisol level. The concentration of <em>17OHP</em> was above 6.0 nmol/l (200 ng/dl) in 41 patients and above 30.2 nmol/l (1,000 ng/dl) in 10. Higher <em>17OHP</em> levels and more severe diseases correlated positively.
CONCLUSIONS
During infectious diseases some patients demonstrated not only cortisol elevation but also <em>17OHP</em> as high as that observed in NC-CAH. We suggest that if <em>17OHP</em> elevation is not characteristic of SL-CAH, glucocorticoid therapy should be started and an ACTH test should be performed after recovery before ruling out this pathology.
Journal: Acta Endocrinologica
February/13/1985
Abstract
The effect of daily injections of D-Ser-(TBU)6-LRH-EA10 (GnRH analogue (GnRH-A) 100 micrograms sc) on serum testosterone (T), 17 alpha-hydroxyprogesterone (<em>17OHP</em>) and oestradiol-17 beta (E2) was studied in 4 men. During GnRH-A therapy T, <em>17OHP</em> and E2 were markedly decreased by the end of the second month. Continuous long-term administration of GnRh-A inhibited testicular function. To test whether the biosynthetic pathway was affected by the regimen, a bolus of 2000 U hCG was given to each subject after 10 months of therapy. Evaluation of the kinetics of steroid responsiveness showed a significant release of T in response to the trophic stimulus, with little or no elevation of serum <em>17OHP</em> and E2. The response seen in these treated men appeared similar to that found in hypogonadotrophic men and prepubertal boys.
Journal: General and Comparative Endocrinology
January/26/2020
Abstract
17α, 20β-Dihydroxy-4-pregnen-3-one (DHP) is a maturation-inducing steroid in many teleost fish. Carbonyl reductase-like 20β-hydroxysteroid dehydrogenase (CR/20β-HSD) is a candidate enzyme responsible for DHP production during oocyte maturation in various fish, including Nile tilapia. However, a novel type of 17β-hydroxysteroid dehydrogenase, type 12-like (17β-HSD12L), is responsible for DHP production during oocyte maturation in masu salmon. 17β-HSD12 (presumably orthologous to salmon 17β-HSD12L) has been detected in Nile tilapia; however, its enzymatic activity and specific ability to convert the DHP substrate 17α-hydroxyprogesterone (<em>17OHP</em>) have not been examined. This study aimed to determine whether CR/20β-HSD or 17β-HSD12 is responsible for DHP production during oocyte maturation in the Nile tilapia. Mammalian expression vectors containing tilapia hsd17b12 or CR/20bhsd were transfected into HEK293T cells, followed by incubation with <em>17OHP</em>. HEK293T cells transfected with hsd17b12 exhibited a strong ability to convert exogenous <em>17OHP</em> to DHP (73.8% yield). Cells transfected with CR/20bhsd or the control vector converted only 7.4% and 7.5% of <em>17OHP</em> to DHP, respectively. In addition, based on LC-MS/MS analyses, 17β-HSD12 did not convert any substrates other than <em>17OHP</em>, including DHP, adrenosterone, androstenedione, estrone, testosterone, 11-ketotestosterone, and estradiol-17β. CR/20β-HSD showed strong 17β-HSD oxidoreductase activity especially with adrenosterone and androstenedione. Tissue-specific hsd17b12 expression analyzed by RT-PCR showed that hsd17b12 mRNA was strongest amplification in full-grown follicles. Finally, full-grown ovarian follicles were incubated with salmon pituitary extract (SPE, 100 µg/mL) or human chorionic gonadotropin (HCG,100 IU/mL) to induce 20β-HSD activity in vitro, and enzyme activity was assessed by co-incubation with 100 ng/mL <em>17OHP</em> for 2, 4, 8, and 16 h. Conversion of <em>17OHP</em> to DHP by ovarian follicles incubated with SPE and HCG peaked at 16 h, subsequent with increased follicular hsd17b12 mRNA levels, which were significantly higher than those in control incubations. However, the levels of CR/20bhsd mRNA remained low and did not differ among time points. The present study strongly suggests that 17β-HSD12, and not CR/20β-HSD, is the 20β-HSD responsible for DHP production by ovarian follicles during oocyte maturation in Nile tilapia.
Journal: Journal of Pediatrics
November/14/2021
Abstract
Objective: (s): To assess whether 21-deoxycortisol (21deoxy) can be used to predict 21-hydroxylase deficiency (21OHD) in newborns and to evaluate the influence of gestational age and the timing of collection on 21-deoxycortisol concentrations which can impact screening specificity.
<strong class="sub-title"> Study design: </strong> 17-hydroxyprogesterone (<em>17OHP</em>) and 21-deoxycortisol (21deoxy) were measured in 906 newborn screening specimens (851 unaffected newborns, 55 confirmed cases of 2OHD) to compare their ability to identify babies with 21-hydroxylase deficiency. Additionally, these two steroids were scrutinized in the unaffected cohort to determine the influence of gestational age (ranging from 23-42 weeks) and the timing of the measured concentrations.
<strong class="sub-title"> Results: </strong> The gestational age of the newborn impacted both <em>17OHP</em> and 21deoxy concentrations, but the degree of influence was more substantial for <em>17OHP</em>. Timing of collection, however, did not affect 21deoxy concentrations. Additionally, 21deoxy was a better predictor of 21OHD status than <em>17OHP</em>, with little overlap in concentrations between the unaffected population and confirmed cases of 21OHD. A streamlined decision tree using solely 21deoxy (cutoff values of 0.85 ng/ml), yielded a 91.7% positive predictive value for 21OHD screening.
Conclusions: Our findings demonstrate that 21deoxy is a key disease marker of 21OHD and could be used to improve the accuracy of newborn screening for this disorder.
Keywords: 17-hydroxyprogesterone; 21-deoxycortisol; Newborn screening; congenital adrenal hyperplasia; liquid chromatography tandem mass spectrometry; second-tier testing.
Related with
Journal: Archives of endocrinology and metabolism
November/10/2021
Abstract
Ovarian adrenal rest tumors (OARTs) are very rare. We describe a case of a young woman with uncontrolled classical congenital adrenal hyperplasia (CCAH), presenting with bilateral OARTs, successfully treated with steroid replacement. A 20-year-old woman, known to have 21OH-CCAH, presented with severe abdominal pain, vomiting, diarrhea, and fever. As a result of poor compliance, 6 months before her admission hirsutism worsened and amenorrhea, hyperpigmentation, and weakness developed. ACTH levels were 278 < pmol/L and <em>17OHP</em> 91.3 nmol/L. She was admitted for parenteral antibiotics and high-dose hydrocortisone treatment. CT revealed bilateral juxta-ovarian masses (6.2 × 3.6 × 7.4 cm left and 5 × 2.2 × 3.2 cm right) that on MRI were iso-intense in T1 and hypointense in T2, with early enhancement and rapid washout. One week of high-dose hydrocortisone resulted in significant clinical and laboratory improvement and the patient was discharged with 2 mg dexamethasone/day. One month later US revealed shrinkage of the masses and dexamethasone dose was decreased. At three months from discharge, she has resumed regular menses, and a repeated MRI revealed the para-ovarian masses have shrunk. One year after the diagnosis, the para-ovarian masses have shrunk more to 2.8 × 1.9 × 4.3 on the left and 2.1 × 0.9 × 1.2 on the right with less contrast enhancement in comparison to previous test possibly due to fibrotic changes of the tissue. OARTs are rare tumors with a poorly known natural history, and surgery has been the first option in the few reported cases. We demonstrate that medical treatment is a good alternative, leading to significant tumor shrinkage over a short period.
Related with
Journal: Scandinavian Journal of Clinical and Laboratory Investigation
November/30/2020
Abstract
Quantitation of endogenous steroids and their precursors is essential for diagnosis of a wide range of endocrine disorders. Usually, these analyses have been carried out using immunoassays. However, immunoassays often overestimate concentrations due to assay interference by other endogenous steroids, especially for low concentrations. Mass spectrometry based methods offer superior specificity, accuracy, and sensitivity. We therefore present a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with automated sample preparation for determination of 17α-hydroxyprogesterone (<em>17OHP</em>), cortisol, cortisone, dehydroepiandrosterone sulfate (DHEAS), androstenedione (A<sub>4</sub>), testosterone (T), and estrone sulfate (E<sub>1</sub>S). Samples were prepared using protein precipitation and 96-well filter plates, fully automated in a pipetting robot and analyzed by LC-MS/MS. Serum samples from 187 healthy children and adolescents aged 5-18 years were used to study hormone changes in relation to sex and pubertal stage. Lower limit of quantification for <em>17OHP</em> was 0.7 nmol/L, for cortisol 11 nmol/L, for cortisone 2 nmol/L, for DHEAS 0.1 µmol/L, and for A<sub>4</sub>, T, and E<sub>1</sub>S, 0.2 nmol/L. This study showed a general increase in <em>17OHP</em>, DHEAS, A<sub>4</sub>, T and E<sub>1</sub>S in both genders during puberty. In boys, A<sub>4</sub> and T increased significantly throughout pubertal development. Girls had significantly higher A<sub>4</sub> and E<sub>1</sub>S concentrations, while boys had higher T concentrations. No sex- or puberty-specific differences were seen in cortisol or cortisone concentrations. To the best of our knowledge, this is the first presentation of changes in serum E<sub>1</sub>S concentrations during pubertal development in healthy children.
Keywords: Androgens; children; cortisol/cortisone ratio; estrone sulfate; mass spectrometry; puberty.
Related with
Journal: Hormones
October/29/2020
Abstract
Introduction: P450 oxidoreductase (POR) deficiency is a rare form of congenital adrenal hyperplasia. In both genders, it can lead to ambiguous genitalia, impaired steroidogenesis, and skeletal findings similar to those of Antley-Bixler syndrome.
<strong class="sub-title"> Cases: </strong> We describe two cases of POR deficiency. The first case was an 8.5-year-old girl who was admitted to our clinic due to ambiguous genitalia. Karyotype was 46, XX. There were mild dysmorphic facial findings and mild metacarpophalangeal joint deformity. The patient's basal cortisol and ACTH levels were normal, while 17-hydroxyprogesterone (<em>17OHP</em>) levels were high. Peak cortisol response to the ACTH stimulation test was found to be insufficient. Our second case, a sibling of the first case, was admitted for routine checkup at the age of 15 months. As in our first case, there were dysmorphic facial findings and metacarpophalangeal joint deformity. The genital structure was normal. Karyotype was 46, XY. Basal cortisol and ACTH levels were normal, while <em>17OHP</em> level was slightly high. Peak cortisol response to the ACTH stimulation test was found to be insufficient. Based on our findings, POR deficiency was considered in both of these cases and NM_000941.3:c.929_937delTCTCGGACT(p.Ile310_Ser313delinsThr) (homozygous) mutation was detected in the POR gene that had not previously been described.
Conclusion: We detected a novel variant in the POR gene in two sibling cases with adrenal insufficiency, dysmorphic face, and mild skeletal findings. While the detected mutation caused ambiguous genitalia in the female case, it did not cause ambiguous genitalia in the male case.
Keywords: Adrenal insufficiency; Congenital adrenal hyperplasia; Disorders of sexual development; POR deficiency; Skeletal malformation.
Related with
Journal: Dermatologic Therapy
November/12/2020
Abstract
Polycystic ovarian syndrome (PCOS) diagnosis in adult female acne (AFA) is tough owing to unreliable ultrasonography in virgins or obese females and inconsistent hyperandrogenemia. We analyzed hormones in AFA and established a diagnostic cut-off value of anti-mullerian hormone (AMH) for PCOS. Female acne patients aged ≥25 years were assessed with total testosterone (TT), sex hormone binding globulin (SHBG), free androgen index (FAI), AMH, 17-hydroxyprogesterone (17-OHP), dehydroepiandrosterone sulfate (DHEAS), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Rotterdam's criteria defined PCOS. AMH was measured (Access AMH assay) to calculate the diagnostic cut off value using receiver operating characteristic (ROC) curve. Of 120 cases, 25.83% had PCOS. This group had significant clinical hyperandrogenism, truncal and adolescent acne, polycystic ovarian morphology (PCOM), and raised hormones (AMH, TT, FAI, LH, and LH/FSH). AMH levels were significantly higher in the PCOS group (6.91 ± 3.85 ng/mL) and positively correlated with TT, FAI, <em>17OHP</em>, LH, and LH/FSH ratio. AMH at >5.1 ng/mL (sensitivity-70.97% and specificity-82.02%) predicted PCOS and correlated with PCOM. AMH (>5.1 ng/mL) is useful for diagnosing PCOS and surrogate for hyperandrogenemia and PCOM. Its correlation with hormones in non PCOS AFA highlights its sensitivity to diagnose endocrinological derangements.
Keywords: AMH; PCOS; adult female acne; biochemical hyperandrogenemia; hyperandrogenism.
Related with
Journal: BMC Endocrine Disorders
November/25/2021
Abstract
Background: Our study aims to summarize the clinical characteristics of rare types of congenital adrenal hyperplasia (CAH) other than 21-hydroxylase deficiency (21-OHD), and to explore the clinical applications of genetic analysis and liquid chromatography tandem-mass spectrometry (LC-MS/MS) in rare CAH.
Methods: We retrospectively analysed the clinical data of 5 rare cases of CAH admitted to our hospital and summarized their clinical manifestations, auxiliary examinations, diagnosis and mutational spectrum.
<strong class="sub-title"> Results: </strong> After gene sequencing, complex heterozygous variants were detected in all patients (2 cases were lipoid congenital adrenal hyperplasia (LCAH), 11β-hydroxylase deficiency (11β-OHD), 3β-hydroxysteroid dehydrogenase deficiency (3β-HSD deficiency) and P450 oxidoreductase deficiency (PORD) each accounted for 1 case), which were consistent with their clinical manifestations. Among them, 4 novel variants were detected, including c.650 + 2 T > A of the StAR gene, c.1145 T > C (p. L382P) of the CYP11B1 gene, c.1622C > T (p. A541V) and c.1804C > T (p. Q602 *) of the POR gene. The LC-MS/MS results for steroid hormones in patients were also consistent with their genetic variants: 2 patients with LCAH showed a decrease in all steroid hormones; 11β-OHD patient showed a significant increase in 11-deoxycortisol and 11-deoxycorticosterone; patient with 3β-HSD deficiency showed a significant increase in DHEA; and PORD patient was mainly characterized by elevated <em>17OHP</em>, progesterone and impaired synthesis of androgen levels.
Conclusions: The clinical manifestations and classification of CAH are complicated, and there are cases of missed diagnosis or misdiagnosis. It's necessary to combine the analysis of clinical manifestations and auxiliary examinations for diagnosis; if necessary, LC-MS/MS analysis of steroid hormones or gene sequencing is recommended for confirming diagnosis and typing.
Keywords: 11β-hydroxylase deficiency; 3β-hydroxysteroid dehydrogenase deficiency; Congenital adrenal hyperplasia; Lipoid congenital adrenal hyperplasia; P450 oxidoreductase deficiency.
Related with
Journal: Biomedical Research
September/2/2021
Abstract
Background: Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder caused by impaired steroidogenesis. Glucocorticoid treatment with increased androgens may lead to cardiovascular and metabolic effects in these patients. In this study, we investigated the relationship between cardiovascular risk factors and androgen levels in children and adolescents with CAH due to 21 hydroxylase deficiency.
<strong class="sub-title"> Materials and methods: </strong> A cross-sectional study of 78 patients (37 boys and 41 girls) with CAH aged 3-17 years. Anthropometric, body mass index (BMI), systolic (SBP), and diastolic (DBP) blood pressure were measured. Fasting blood glucose with plasma insulin and lipids were measured, and insulin resistance (HOMA-IR) calculated using the homeostasis assessment model. Furthermore, testosterone, Dehydroepiandrosterone sulfate (DHEAS), and 17-Hydroxyprogesterone (<em>17OHP</em>) were investigated.
<strong class="sub-title"> Results: </strong> The mean SBP and DBP were 112.01 ± 19.13 and 69.77 ± 7.56, respectively. The mean of HOMA-IR in patients was 2.25 ± 1.46. The frequency of patients with overweight and High HOMA index were, respectively, 33.3% and 29.3%. The correlation analysis between clinical characteristics and androgen serum levels showed that DBP and BMI had a significant positive correlation with <em>17OHP</em>. The median regression analysis showed, only DBP in the adjusted model had a significant positive effect with <em>17OHP</em> level (<i>P</i> < 0.05), and no significant relationship was observed for other characteristics.
Conclusion: A significant association was found between BMI and DBP with serum concentrations of 17-OHP, suggesting that elevated 17-OHP can lead to an increased risk of cardiovascular disorders in children and adolescents with CAH.
Keywords: 17-hydroxyprogesterone; Cardiovascular System; androgens; blood pressure; congenital adrenal hyperplasia.
Journal: Journal of Maternal-Fetal and Neonatal Medicine
September/1/2021
Abstract
<strong class="sub-title"> Objective: </strong> The aim of this study was to compare the effect of vaginal progesterone with 17-alpha-hydroxyprogesterone caproate (<em>17OHP</em>-C) in prevention of preterm birth in high-risk pregnant women undergo cerclage.
<strong class="sub-title"> Materials and methods: </strong> This prospective randomized clinical trial registered in the Iranian Registry of Clinical Trials (IRCT20181107041585N4), was performed from May 2017 to August 2018 in Bandar Abbas, Iran. Fifty-eight eligible women who were scheduled for cervical cerclage due to a history of two or more previous preterm birth <28 weeks or a cervical length less than 25 mm with at least one previous preterm birth <34 weeks were randomly divided into two groups. The first group received 200 mg of vaginal progesterone suppository daily and the second one received 250 mg of <em>17OHP</em>-C intramuscular weekly after cerclage procedure until the end of 36 weeks. Patients were followed up to the end of delivery and the newborn until the first 28 d after delivery.
<strong class="sub-title"> Results: </strong> Gestational age at the time of birth in <em>17OHP</em>-C group was significantly higher than vaginal progesterone group (<i>p</i>=.021). However, the incidence of preterm birth in both groups was not statistically significant (20.7% <i>vs.</i> 24.1%). Apgar scores, newborn birthweight, admission to neonatal intensive care unit (NICU), incidence of respiratory distress syndrome (RDS), sepsis, necrotizing enterocolitis (NEC), and, intraventricular hemorrhage (IVH), was similar in both groups. Adverse events were reported in 48.3% of patients in 17-OHP-C group, and 27.6% of patients in the vaginal progesterone group (<i>p</i>= .014).
<strong class="sub-title"> Conclusions: </strong> Vaginal progesterone and <em>17OHP</em>-C had similar results in terms of prevention of preterm birth and neonatal outcome. However, the adverse events associated with 17-OHP-C were higher than vaginal progesterone.
Keywords: 17-Alpha-hydroxyprogesterone caproat; cerclage; preterm birth; vaginal progesterone[AQ].
Journal: Journal of the Endocrine Society
December/7/2021
Abstract
Ovarian hyperthecosis (OHT), severe hyperandrogenism after menopause in the absence of ovarian or adrenal tumors, is usually treated by surgical excision. We report a 58-year-old woman presenting with severe hyperandrogenism (serum testosterone 15.7-31.0 nmol/L, normal female <1.8 nmol/L) with menopausal gonadotropins and virilization but no adrenal or ovarian lesions. Multisteroid profiling by liquid chromatography mass spectrometry (LCMS) of adrenal and ovarian vein samples identified strong gradients in the left ovarian vein (10- to 30-fold vs peripheral blood in <em>17OHP</em><sub>4</sub>, 17 hydroxyprogesterone, 17 hydroxypregnenolone, androstenedione, testosterone, dehydroepiandrosterone) but the right ovarian vein could not be cannulated with the same findings in a second ovarian vein cannulation. OHT diagnosis was confirmed by an injection of a depot pure gonadotropin-releasing hormone (GnRH) antagonist (80 mg Degarelix, Ferring) producing a rapid (<24 hour) and complete suppression of ovarian steroidogenesis as well as serum luteinizing hormone and follicle-stimulating hormone lasting at least 8 weeks, with reduction in virilization but injection site reaction and flushing and vaginal spotting ameliorated by an estradiol patch. Serum testosterone remained suppressed at 313 days after the first dose despite recovery of menopausal gonadotropins by day 278 days. This illustrates use of multisteroid LCMS profiling for confirmation of the OHT diagnosis by ovarian and adrenal vein sampling and monitoring of treatment by peripheral blood sampling. Injection of a depot pure GnRH antagonist produced rapid and long-term complete suppression of ovarian steroidogenesis maintained over 10 months. Hence a depot pure GnRH antagonist can not only rapidly confirm the OHT diagnosis but also induce long-term remission of severe hyperandrogenism without surgery.
Keywords: GnRH antagonist; liquid chromatography mass spectrometry; ovarian hyperthecosis; ovarian vein sampling; steroid profile; testosterone.
A Comparison of Vaginal and Intramuscular Progesterone for the Prevention of Recurrent Preterm Birth
Journal: American Journal of Perinatology
December/13/2021
Abstract
<strong class="sub-title"> Objective: </strong> To examine whether vaginal progesterone is noninferior to 17-α hydroxyprogesterone caproate (<em>17OHP</em>-C) in the prevention of recurrent preterm birth (PTB).
Study design: This retrospective cohort study included singleton pregnancies among women with a history of spontaneous PTB who received prenatal care at a single tertiary center from 2011 to 2016. Pregnancies were excluded if progesterone was not initiated prior to 24 weeks or the fetus had a major congenital anomaly. The primary outcome was PTB <37 weeks. A priori, noninferiority was to be established if the upper bound of the adjusted two-sided 90% confidence interval (CI) for the difference in PTB fell below 9%. Inverse probability of treatment weighting (IPTW) was used to carefully control for confounding associated with choice of treatment and PTB. Adjusted differences in PTB proportions were estimated via IPTW regression, with standard errors adjustment for multiple pregnancies per woman. Secondary outcomes included PTB <34 and <28 weeks, spontaneous PTB, neonatal intensive care unit admission, and gestational age at delivery.
<strong class="sub-title"> Results: </strong> Among 858 pregnancies, 41% (<i>n</i> = 353) received vaginal progesterone and 59% (<i>n</i> = 505) were given <em>17OHP</em>-C. Vaginal progesterone use was more common later in the study period, and among women who established prenatal care later, had prior PTBs at later gestational ages, and whose race/ethnicity was neither non-Hispanic white nor non-Hispanic black. Vaginal progesterone did not meet noninferiority criteria compared with 17-OHPC in examining PTB <37 weeks, with an IPTW adjusted difference of 3.4% (90% CI: -3.5, 10.3). For secondary outcomes, IPTW adjusted differences between treatment groups were generally small and CIs were wide.
<strong class="sub-title"> Conclusion: </strong> We could not conclude noninferiority of vaginal progesterone to <em>17OHP</em>-C; however, women and providers may be willing to accept a larger difference (>9%) when considering the cost and availability of vaginal progesterone versus <em>17OHP</em>-C. A well-designed randomized trial is needed.
<strong class="sub-title"> Key points: </strong> · Vaginal progesterone is not noninferior to <em>17OHP</em>-C.. · PTB risk may be 10% higher with vaginal progesterone.. · Associations did not differ based on obesity status..
Journal: Endocrinology and Metabolism
October/14/2021
Abstract
Context: Classic congenital adrenal hyperplasia due to 21-hydroxylase deficiency (21OHD) is characterized by impaired cortisol synthesis and excess androgen production. Corticotropin-releasing factor type 1 receptor (CRF1R) antagonism may decrease adrenal androgen production.
Objective: To evaluate the safety, tolerability, and efficacy of crinecerfont (NBI-74788), a selective CRF1R antagonist, in 21OHD.
Design: Open-label, phase 2 study, with sequential cohort design (NCT03525886).
Setting: United States (6 centers).
Participants: Men and women, 18-50 years, with 21OHD.
Interventions: Four crinecerfont regimens, each administered orally for 14 consecutive days: 50 or 100 mg oncedaily at bedtime (Cohorts 1 and 2, respectively); 100 mg once-daily in the evening (Cohort 3); 100 mg twice-daily (BID, Cohort 4). Participants could enroll in >1 cohort.
<strong class="sub-title"> Main outcomes: </strong> Changes from baseline to Day 14 in adrenocorticotropic hormone (ACTH), 17hydroxyprogesterone (<em>17OHP</em>), androstenedione, and testosterone.
<strong class="sub-title"> Results: </strong> Eighteen participants (11 women, 7 men) were enrolled: Cohort 1 (n=8), Cohort 2 (n=7), Cohort 3 (n=8), Cohort 4 (n=8). Mean age was 31 years; 94% were white. Median percent reductions were >60% for ACTH (-66%), <em>17OHP</em> (64%), and androstenedione (64%) with crinecerfont 100 mg BID. In female participants, 73% (8/11) had ≥50% reduction in testosterone levels; male participants had median 26-65% decreases in androstenedione/testosterone ratios.
<strong class="sub-title"> Conclusions: </strong> Crinecerfont treatment for 14 days lowered ACTH and afforded clinically meaningful reductions of elevated <em>17OHP</em>, androstenedione, testosterone (women), or androstenedione/testosterone ratio (men) in adults with 21OHD. Longer-term studies are required to evaluate the effects of crinecerfont on clinical endpoints of disordered steroidogenesis and glucocorticoid exposure in patients with 21OHD.
Keywords: 17-hydroxyprogesterone; 21-hydroxylase deficiency; NBI-74788; congenital adrenal hyperplasia; crinecerfont.
Related with
Journal: Endocrinology and Metabolism
October/24/2021
Abstract
Objective: Insulin sensitivity evaluation by hyperinsulinemic-euglycaemic clamp in non-classical congenital adrenal hyperplasia (NC-CAH) due to 21-hydroxilase deficiency.
Design and setting: Cross-sectional study at University Hospital outpatient clinics.
Patients and methods: NC-CAH patients (25F/5M; 24 ± 10 years) subdivided into C/NC (compound heterozygous for one classical and one non-classical allele) and NC/NC (two non-classical alleles) genotypes were compared to controls.
<strong class="sub-title"> Results: </strong> At diagnosis, C/NC patients presented higher basal and ACTH-stimulated <em>17OHP</em> and androstenedione levels than NC/NC genotype. Patients and controls presented similar weight, BMI, abdominal circumference, and total fat body mass. NC-CAH patients showed higher waist-to-hip ratio, lower adiponectin and HDL cholesterol levels with no changes in fasting plasma glucose, HbA1c, HOMA-IR, adiponectin, leptin, IL-6, TNF-alfa, CRP, and carotid-IMT. All patients had used glucocorticoid (mean time of 73 months). Among the twenty-two patients with successful clamp, thirteen were still receiving glucocorticoid - three patients using cortisone acetate, nine dexamethasone, and one prednisone (hydrocortisone equivalent dose of 5.5 mg/m²/day), while nine patients were off glucocorticoid but had previously used (hydrocortisone equivalent dose of 5.9 mg/m 2/day). The NC-CAH patients presented lower Mffm than controls (31±20 vs 55±23 µmol.min -1.kg -1, p=0.002). The Mffm values were inversely correlated with the duration of glucocorticoid treatment (r=-0.44, p=0.04). There was association of insulin resistance and glucocorticoid type but not with androgen levels.
Conclusion: Using the gold standard method, the hyperinsulinemic-euglycemic clamp, insulin resistance was present in NC-CAH patients and related to prolonged use and long-acting glucocorticoid treatment. Glucocorticoid replacement and cardiometabolic risks should be monitored regularly in NC-CAH.
Keywords: genotype; hyperinsulinemic-euglycaemic clamp; insulin resistance; nonclassical congenital adrenal hyperplasia.
Journal: Frontiers in Endocrinology
February/9/2022
Abstract
<strong class="sub-title"> Background: </strong> Biochemically monitoring 21-hydroxylase deficiency (21-OHD) is challenging. Serum/blood 17-hydroxyprogesterone (<em>17OHP</em>) measurements are normally used for this purpose. Urinary pregnanetriol (PT), a urinary metabolite of <em>17OHP</em>, may also be used. Based on auxological data, we previously reported that the optimal first morning PT value fell in the range of 2.2-3.3 mg/gCr (95% confidence interval of the mean) and 0.59-6.0 mg/gCr (10<sup>th</sup> - 90<sup>th</sup> percentile) for monitoring 21-OHD treatment. No report thus far has directly compared the first morning urinary PT value with the <em>17OHP</em> value at various times during the day.
<strong class="sub-title"> Objective: </strong> To explore the correlation between the first morning urinary PT value before glucocorticoid administration and the serum/blood <em>17OHP</em> value at three time points, namely, before and two and four hours after glucocorticoid administration.
Design: This was a prospective study done at two children's hospitals.
<strong class="sub-title"> Methods: </strong> In total, 25 patients with 21-OHD aged 3-25 years were recruited. Their urinary PT levels and <em>17OHP</em> levels were measured for three days within a total period of one week. The first morning PT value was collected on all three days. Dried blood spots and serum were used to measure <em>17OHP</em>.
<strong class="sub-title"> Results: </strong> The range for the first morning PT value for all the samples (n=69) was 0.10-56.1 mg/gCr. A significant, positive correlation was found between the first morning PT and <em>17OHP</em> values before medication (r=0.87, p<0.01), and weaker correlation was observed between the first morning PT and <em>17OHP</em> values after medication.
<strong class="sub-title"> Conclusions: </strong> The first morning PT correlated more significantly with <em>17OHP</em> before the morning medication. Measuring the first morning PT value may be more practical and useful for monitoring 21-OHD biochemically.
Keywords: 17-hydroxyprogesterone; 21-hydroxylase deficiency; Urinary pregnanetiol; congenital adrenal hyperplasia; first morning urine sample; therapy monitoring, glucocorticoid.
Correlation between morning PT and 17OHP. (A) Morning PT and morning DBS 17OHP showed a significant...Related with
Results with error correction
Journal: Clinical Endocrinology
October/15/1997
Abstract
OBJECTIVE
Polycystic ovary syndrome (PCOS) is a common endocrinopathy of unknown aetiology. The aims of this study were to identify whether ovarian thecal cell steroidogenesis is abnormally regulated in PCOS by measuring steroid responses to a single dose of hCG before, and during, suppression of endogenous LH levels by GnRH analogue (GnRHa).
METHODS
Serum levels of LH, FSH, 17 alpha hydroxyprogesterone (<em>17OHP</em>), androstenedione, testosterone and dehydroepiandrosterone sulphate were measured before, and 48 hours after, a single intramuscular injection of 10,000 IU hCG. The test was repeated 4 weeks after suppression of endogenous LH levels by GnRHa.
METHODS
The ovarian responses to hCG were compared in three groups of women. Eleven women had normal ovaries and regular cycles, eight had polycystic ovaries but not clinical features of the syndrome (PCO group) and eight had polycystic ovaries, anovulation and either severe hirsutism or alopecia (PCOS group).
RESULTS
Before GnRHa treatment, LH levels were significantly higher in the PCOS group but hCG stimulated a similar rise in <em>17OHP</em> in all three groups. Following analogue, LH levels were suppressed in all three groups but the <em>17OHP</em> responses to hCG were significantly higher in both the PCO and PCOS groups compared with normal controls.
CONCLUSIONS
These findings provide further evidence in favour of an intrinsic abnormality of thecal cell steroidogenesis in the polycystic ovary.
Journal: Fertility and Sterility
October/10/2001
Abstract
OBJECTIVE
To examine the direct effect of metformin on thecal cell androgen production.
METHODS
Basic science research laboratory, University of Texas Southwestern, Dallas, Texas.
METHODS
Human ovarian theca-like tumor cells were treated with various concentrations of metformin in the presence and absence of forskolin for 48 hours.
METHODS
Media were collected, and radioimmunoassay (RIA) for progesterone, 17 alpha-hydroxyprogesterone (<em>17OHP</em>), androstenedione, and testosterone was performed. The effect of metformin on the expression of various enzymes involved in theca cell steroidogenesis was examined.
RESULTS
Metformin (50 microM and 200 microM) significantly inhibited androstenedione production from both forskolin-stimulated and unstimulated theca cells. Testosterone production was also significantly inhibited in forskolin-treated cells in the presence of 200 microM of metformin-treated compared with forskolin-only-treated cells. Western blot analysis revealed that metformin significantly inhibited the expression of steroidogenic acute regulatory (StAR) protein and 17 alpha-hydroxylase (CYP17) expression in cells stimulated with forskolin compared with forskolin treatment alone. There was no significant change in either 3beta-hydroxysteroid dehydrogenase (3 beta HSD) or cholesterol side-chain cleavage (CYP11A1) protein expression. Northern analysis revealed a significant decrease in the expression of CYP17 mRNA in forskolin-stimulated cells treated with metformin (200 microM) compared with forskolin-only-treated cells, however, there was no significant change in steroidogenic acute regulatory protein mRNA expression.
CONCLUSIONS
Our results suggest that metformin may have a direct effect on thecal cells' androgen production.
Journal: European Journal of Endocrinology
January/4/2005
Abstract
Congenital adrenal hyperplasia (CAH) is well suited for newborn screening, as it is a common and potentially fatal disease which can be easily diagnosed by a simple hormonal measurement in blood. Moreover, early recognition and treatment can prevent severe salt wasting, dehydration and death and shorten the time of male sex assignment in virilised females.In screening programmes, 17alpha-hydroxyprogesterone (<em>17OHP</em>) is measured in filter paper blood spots obtained by a heel puncture preferably between 2 and 4 days after birth. Three assay techniques are utilised for initial screening: radio-immunoassay (USA), enzyme-linked immunosorbent assay (Japan) and time-resolved fluoro-immunoassay (Europe). Preterm newborns have higher <em>17OHP</em> concentrations in serum than babies born at term. Therefore, cut-off levels are based on gestational age (in Japan and Europe) or on birth weight (in the USA). There is a considerable variation in cut-off levels from one programme to another. This is most likely due to the different antibodies and reagents used, varying thickness and density of filter paper used for sample collection and, most significantly, the characteristics of the reference population (in terms of birth weight and gestational age). More than 30 million newborns have been screened. The prevalence of CAH in the USA and Europe is approximately 1:15 000-16 000, and slightly lower in Japan (1:19 000). In general, severe salt wasting can be prevented, but there is a remarkable variation in the number of false-positives and false-negatives among the various programmes. Ongoing refinement of cut-off levels is needed to improve specificity and sensitivity.
Journal: Journal of Biological Chemistry
May/15/2012
Abstract
Steroid 21-hydroxylase (cytochrome P450 21A2, CYP21A2) deficiency accounts for ∼95% of individuals with congenital adrenal hyperplasia, a common autosomal recessive metabolic disorder of adrenal steroidogenesis. The effects of amino acid mutations on CYP21A2 activity lead to impairment of the synthesis of cortisol and aldosterone and the excessive production of androgens. In order to understand the structural and molecular basis of this group of diseases, the bovine CYP21A2 crystal structure complexed with the substrate 17-hydroxyprogesterone (<em>17OHP</em>) was determined to 3.0 Å resolution. An intriguing result from this structure is that there are two molecules of <em>17OHP</em> bound to the enzyme, the distal one being located at the entrance of the substrate access channel and the proximal one bound in the active site. The substrate binding features locate the key substrate recognition residues not only around the heme but also along the substrate access channel. In addition, orientation of the skeleton of the proximal molecule is toward the interior of the enzyme away from the substrate access channel. The <em>17OHP</em> complex of CYP21A2 provides a good relationship between the crystal structure, clinical data, and genetic mutants documented in the literature, thereby enhancing our understanding of congenital adrenal hyperplasia. In addition, the location of certain CYP21A2 mutations provides general understanding of structure/function relationships in P450s.
Journal: Gynecological Endocrinology
July/14/2008
Abstract
OBJECTIVE
To evaluate the effects the administration of myo-inositol (MYO) on hormonal parameters in a group of PCOS patients.
METHODS
Controlled clinical study.
METHODS
PCOS patients in a clinical research environment.
METHODS
20 overweight PCOS patients were enrolled after informed consent.
METHODS
All patients underwent hormonal evaluations and an oral glucose tollerance test (OGTT) before and after 12 weeks of therapy (Group A (n = 10): myo-inositol 2 gr. plus folic acid 200 mug every day; Group B (n = 10): folic acid 200 mug every day). Ultrasound examinations and Ferriman-Gallwey score were also performed.
METHODS
Plasma LH, FSH, PRL, E2, <em>17OHP</em>, A, T, glucose, insulin, C peptide concentrations, BMI, HOMA index and glucose-to-insulin ratio.
RESULTS
After 12 weeks of MYO administration plasma LH, PRL, T, insulin levels and LH/FSH resulted significantly reduced. Insulin sensitivity, expressed as glucose-to-insulin ratio and HOMA index resulted significantly improved after 12 weeks of treatment. Menstrual cyclicity was restored in all amenorrheic and oligomenorrheic subjects. No changes occurred in the patients treated with folic acid.
CONCLUSIONS
Myo-inositol administration improves reproductive axis functioning in PCOS patients reducing the hyperinsulinemic state that affects LH secretion.
Journal: Journal of Clinical Endocrinology and Metabolism
October/31/2007
Abstract
BACKGROUND
In male patients with congenital adrenal hyperplasia (CAH), testicular adrenal rest tumors are frequently found that may interfere with gonadal function.
OBJECTIVE
Our objective was to determine steroid-producing features of testicular adrenal rest tumors.
METHODS
The study is descriptive and took place at a university medical center.
METHODS
Eight adult CAH patients with bilateral testicular adrenal rest tumors were treated with testis-sparing surgery.
METHODS
In all but one patient, spermatic veins were cannulated during surgery and blood samples collected to measure the adrenal-specific steroid 21-deoxycortisol (21DF) and 17-hydroxyprogesterone (<em>17OHP</em>) and androstenedione (A). The same parameters were measured in simultaneously taken peripheral blood. mRNA concentrations of adrenal-specific enzymes CYP11B1 and CYP11B2 and ACTH and angiotensin II (AII) receptors were measured in tumor tissue.
METHODS
Adrenal-specific steroids/enzymes were assessed.
RESULTS
21DF, <em>17OHP</em>, and A levels were measurable in all spermatic vein samples. The ratio (mean +/- SD) between spermatic vein and simultaneously taken peripheral blood samples was 37.8 +/- 56.3 (21DF), 132.0 +/- 249 (<em>17OHP</em>), and 57.0 +/- 68.2 (A). CYP11B1, CYP11B2, and ACTH and AII receptor mRNAs were detected in all tumors with a strong correlation between ACTH receptor mRNA in tumors and 21DF (r = 0.85; P = 0.015), <em>17OHP</em> (r = 1; P = 0.01) and A (r = 0.89; P = 0.007) concentrations in peripheral blood.
CONCLUSIONS
Testicular adrenal rest tumors produce adrenal-specific steroids and express adrenal-specific enzymes and ACTH and AII receptors, confirming the strong resemblance with adrenal tissue. Because AII receptors are present in tumor tissue, it can be hypothesized that AII may be an additional factor responsible for testicular adrenal rest tumor growth.
Journal: Steroids
June/22/2011
Abstract
BACKGROUND
The simultaneous, rapid and reliable measurement of a wide steroid panel is a powerful tool to unravel physiological and pathological hormone status. Clinical laboratories are currently dominated by high-throughput immunoassays, but these methods lack specificity due to cross-reactivity and matrix interferences. We developed and validated an isotopic dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method for the simultaneous measurement of cortisol, corticosterone, 11deoxycortisol, androstenedione, deoxycorticosterone (DOC), testosterone, 17OHprogesterone, dehydroepiandrosterone (DHEA) and progesterone in serum, and compared it to routine immunoassays employed in our laboratory. We also established adult reference intervals in 416 healthy subjects.
METHODS
0.9 ml of serum were spiked with labelled internal standards (IS) and extracted on C18 cartridges. Eluate was injected into a two-dimensional LC-system, purified in a perfusion column and separated on a C8 column during a 21 min gradient run. Analytes were revealed by atmospheric pressure chemical ionization (APCI) followed by multiple reaction monitoring (MRM) analysis.
RESULTS
Of the four immunoassays compared with the ID-LC-MS/MS method, only the results of ElecsysE170 for cortisol, testosterone in males and progesterone>1 ng/ml were in agreement with ID-LC-MS/MS. ElecsysE170 for testosterone in females and progesterone<1 ng/ml, Immulite2000 for androstenedione, DSL-9000 for DHEA and <em>17OHP</em> Bridge for 17OHprogesterone, respectively, showed poor agreement. Reference intervals and steroid age and fertility related fluctuations were established.
CONCLUSIONS
Our ID-LC-MS/MS method proved to be reliable and sensitive in revealing steroid circulating concentrations in adults and in highlighting the limits of routine immunoassays at low concentrations.
Pulse
Views:
1
Posts:
No posts
Not rated
Journal: Journal of Clinical Endocrinology and Metabolism
July/1/1999
Abstract
Recent reports indicate that girls with premature adrenarche are at risk of developing functional ovarian hyperandrogenism and polycystic ovarian syndrome (PCOS). As insulin and insulin-like growth factors (IGFs) have been implicated in the pathogenesis of PCOS, we hypothesize that they may also have a role in the hyperandrogenism of premature adrenarche. Thirty-five prepubertal girls (23 Caribbean Hispanics and 12 Black African-Americans) underwent a 60-min ACTH and LH-releasing hormone test. Insulin sensitivity (S(I)) was assessed using the frequently sampled i.v. glucose tolerance test with tolbutamide. Fasting levels of IGF-I, IGF-binding protein-1 (IGFBP-1), IGFBP-3, sex hormone-binding globulin, and free testosterone (T) were also obtained. The mean age of the patients was 6.8 yr, and bone age was 8.0 yr. Twenty-five patients had a family history of noninsulin-dependent diabetes mellitus and 19 patients had acanthosis nigricans. The mean S(I) for the entire group was 6.78 +/- 5.21 x 10(-4) min/microU x mL (normal prepubertal S(I), 6.5 +/- 0.54 x 10(-4) min(-1) x microU(-1) x mL(-1)). However, 15 of the 35 girls had an S(I) that was more than 2 SD below the mean reported for normal prepubertal children. Of these 15 patients, 13 were obese, and 14 had acanthosis nigricans. For the entire group of girls, the mean ACTH-stimulated levels of 17-hydroxypregnenolone (<em>17OHP</em>reg), dehydroepiandrosterone (DHEA), androstenedione (AS), 17-hydroxyprogesterone (<em>17OHP</em>), and T and the ACTH-stimulated ratios of <em>17OHP</em>reg/<em>17OHP</em>, <em>17OHP</em>reg/DHEA, <em>17OHP</em>/AS, and DHEA/AS did not differ from the levels reported for Tanner stage II-III pubertal girls. The girls were divided into two groups based on their S(I) (group I, S(I) >2 SD below the mean for age; group II, normal S(I)). The group I girls with a reduced S(I) had significantly higher ACTH-stimulated levels of <em>17OHP</em>reg (group I, 760 +/- 87.84 ng/dL; group II, 428.9 +/- 46.28 ng/dL; P = 0.002), <em>17OHP</em>reg/<em>17OHP</em> ratio (group I, 3.95 +/- 0.36; group II, 2.96 +/- 0.35; P = 0.05), <em>17OHP</em>reg/DHEA (group I, 2.06 +/- 0.21; group II, 1.4 +/- 0.13; P = 0.01), and free T (group I, 1 +/- 0.23 ng/dL; group II, 0.49 +/- 0.19 ng/dL; P = 0.014). Levels of sex hormone-binding globulin were lower in the group I girls. Furthermore, for the entire group of girls, the S(I) correlated inversely with ACTH-stimulated levels of <em>17OHP</em>reg, DHEA, and AS and the ACTH-stimulated ratio of <em>17OHP</em>reg/<em>17OHP</em>. IGF-I correlated inversely with S(I) (r = -0.94; P < 0.001) and correlated directly with the ACTH-stimulated levels of <em>17OHP</em>reg (r = 0.8; P < 0.001) and AS (r = 0.63; P < 0.05). IGF-I also correlated with the ACTH-stimulated ratios of <em>17OHP</em>reg/<em>17OHP</em> (r = 0.61; P < 0.05), <em>17OHP</em>reg/DHEA (r = 0.9; P < 0.001), <em>17OHP</em>/AS (r = 0.79; P < 0.001), and DHEA/AS (r = 0.96; P < 0.001). IGFBP-1 correlated inversely with the ACTH-stimulated levels of <em>17OHP</em>reg (r = -0.38; P < 0.05) and DHEA (r = -0.36; P < 0.05). To summarize, the ACTH-stimulated delta5-steroid levels were higher in prepubertal girls with premature adrenarche and reduced S(I). There was a significant inverse correlation among ACTH-stimulated hormone levels, S(I), and IGFBP-1, whereas IGF-I correlated directly with ACTH-stimulated androgens. These findings support the hypothesis that insulin and IGFs may have a role in the hyperandrogenism of premature adrenarche just as they do in PCOS. Hence, in certain girls with premature adrenarche, hyperandrogenism may be the first presentation of PCOS and/or insulin resistance.
Journal: Journal of Clinical Endocrinology and Metabolism
August/8/2005
Abstract
OBJECTIVE
In newborn screening programs for congenital adrenal hyperplasia, 17-alpha-hydroxyprogesterone (<em>17OHP</em>) cutoff levels are based on birth weight (BW) or on gestational age (GA). We investigated which approach would result in the greatest specificity and sensitivity.
METHODS
For the determination of <em>17OHP</em>, a neonatal <em>17OHP</em> assay was used in filter paper blood of 9492 newborns. The relationships between <em>17OHP</em> and BW and between <em>17OHP</em> and GA were studied by regression analysis. Reference curves with a specificity of 99.95% were constructed with the method that summarizes the distribution by three smoothed curves representing the skewness (L curve), the median (M curve), and the coefficient of variation (S curve). Median cutoff levels for BW and for GA according to the 99.95% reference curves were calculated.
RESULTS
Regression analysis showed that GA is a better predictor of <em>17OHP</em> than BW (R(2) was 50.6 vs. 35.8%, respectively). At a specificity of 99.95%, the calculated median <em>17OHP</em> cutoff level was lower for GA [12.6 microg/liter (38 nmol/liter)] than for BW [17.6 microg/liter (54 nmol/liter)], thus leading to a greater sensitivity.
CONCLUSIONS
This study demonstrates that GA is a better predictor of <em>17OHP</em> in newborns and will result in greater specificity than BW despite the fact that the determination of GA might be less reliable than BW.
Journal: Steroids
January/14/2002
Abstract
We studied influences of dental care, food and storage on the reproducibility of salivary steroid levels. Cortisol (F), 17OH-progesterone (<em>17OHP</em>) and Progesterone (P) were measured using adapted commercial radioimmunoassays. Saliva samples of healthy adults (n = 15; m:8; f:7) were collected directly before and after dental care, and directly before and after breakfast with various foodstuffs. A second experiment investigated stability of steroids under different storage conditions. Four series of identical saliva portions (I: Native saliva; II: Centrifuged saliva; III: Saliva with trifluor acetate (TFA); IV: Saliva with 0.5% NaN(3)) were stored at room temperature and at 4 degrees C for up to three weeks. To demonstrate influences of repeated thawing and re-freezing of saliva on steroid values, saliva samples (n = 15) were divided into identical portions. These portions were frozen and re-thawed up to 5 times before measurement. Neither dental care nor intake of bread or milk effected the reproducibility of F, 170HP, and P. Steroid levels decreased significantly in the course of three weeks under different storage conditions (P < 0.001). This decrease was clinically relevant from the second week onward, with exception of NaN(3) treated samples. After repeated freezing and re-thawing <em>17OHP</em> and P decreased slightly (about 5%). Only F decreased significantly after the third thawing (P < 0.001). The results show the usefulness of standardized handling of saliva samples for improving reproducibility and reliability of salivary steroid measurements.
Journal: Journal of Clinical Endocrinology and Metabolism
May/27/2009
Abstract
OBJECTIVE
Our objective was to determine the ovarian function of asymptomatic volunteers with a polycystic ovary (V-PCO).
METHODS
Non-hirsute eumenorrheic V-PCO (n = 32) and volunteers with ultrasonographically normal ovaries (V-NO) (n = 21) were compared with one another and with polycystic ovary syndrome (PCOS) patients who met National Institute of Health criteria (n = 90). DESIGN/SETTING/INTERVENTIONS: GnRH agonist (GnRHag), ACTH, and oral glucose tolerance tests were prospectively performed in a General Clinical Research Center.
RESULTS
The distribution of 17-hydroxyprogesterone (<em>17OHP</em>) responses to GnRHag of V-PCO formed a distinct population intermediate between that of V-NO, the reference population, and PCOS. Nevertheless, the V-PCO population was heterogeneous. There were 53% (seventeen of 32) that were functionally normal, with <em>17OHP</em> responses and free testosterone levels like V-NO. A total of 25% (eight of 32) had an elevated free testosterone, thus meeting Rotterdam criteria for PCOS; one third of these had <em>17OHP</em> hyperresponsiveness to GnRHag testing. The remaining 22% (seven of 32) had <em>17OHP</em> hyperresponsiveness to GnRHag, but normal free testosterone. Of PCOS, 69% had elevated <em>17OHP</em> hyperresponsiveness to GnRHag. Ovarian volume correlated significantly with <em>17OHP</em> responses only in PCOS, accounting for just 10% of the variance.
CONCLUSIONS
Many asymptomatic volunteers have a PCO. They are a distinct, but heterogeneous, population with respect to ovarian function, ranging from normal (53%) to occult PCOS by Rotterdam criteria (25%). Nearly one quarter (22%) had the typical PCOS type of ovarian dysfunction without hyperandrogenemia, termed a "dysregulated PCO"; they or their offspring may be at risk for PCOS. Ovarian ultrasonographic characteristics must be considered when establishing norms for ovarian function.
Journal: Journal of Steroid Biochemistry and Molecular Biology
May/3/2012
Abstract
In order to overcome many limitations of immunoassays, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has the potential to find its place in the clinical laboratory medicine for quantification of steroid hormones. A prerequisite for the application of a new analytical procedure in clinical diagnostics is standardization to minimize analytical intra- and interlaboratory variability and inaccuracy. We evaluate a newly standardized HPLC-MS/MS assay in kit-format, developed for routine determination of 16 steroid hormones in human serum samples. Fifteen metabolites can be measured quantitatively, which include aldosterone, androstenedione, androsterone, corticosterone, cortisol, cortisone, 11-deoxycortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), 17β-estradiol (E2), estrone (E1), etiocholanolone, 17α-hydroxyprogesterone (<em>17OHP</em>), progesterone, and testosterone. 11-Deoxycorticosterone is the only compound rated as semi-quantitative in this kit. The sample preparation is performed by solid phase extraction (SPE) on a 96-well plate. The standardized assay has been validated for human serum in terms of lower and upper limit of quantification (LLOQ 0.01-32 ng/mL, ULOQ 5-8000 ng/mL), linear correlation coefficient of calibration (R(2)>0.9966), intra- and inter-day precision (intra-day 1.1-8.8%, inter-day 5.2-14.8% and 8.2-18.6% for 11-deoxycorticosterone), accuracy (intra-day 88.3-115.5% and 109.3-128.2% for 11-deoxycorticosterone, inter-day 91.4-117.2% and 102.3-137.1% for 11-deoxycorticosterone), analytical total error (3.6-17.8%), proficiency test accuracy (85.4-113.4%), recovery (68-99%), and metabolite stability (freeze/thaw stability 95.5-108.1%, short term stability 86.9-107.2%). Inter-assay comparison with a routine reference HPLC-MS/MS assay and seven immunoassays demonstrates the outstanding high performance of this HPLC-MS/MS based kit by improvements in accuracy for progesterone, androstenedione, and <em>17OHP</em>. Finally, results of two metyrapone tests demonstrate the potential of the standardized HPLC-MS/MS assay for the analysis of a comprehensive steroid hormone profile in clinical diagnostics.
Related with
Pulse
Views:
1
Posts:
No posts
Not rated
Journal: Journal of Clinical Endocrinology and Metabolism
February/11/2009
Abstract
BACKGROUND
21-Hydroxylase deficiency (21OHD) is caused by CYP21A2 gene mutations disrupting the adrenal 21-hydroxylase, P450c21. CYP21A2 mutations generally correlate well with the 21OHD phenotype, but some children with severe CYP21A2 mutations have residual 21-hydroxylase activity. Some hepatic P450 enzymes can 21-hydroxylate progesterone, but their physiological relevance in modifying 21OHD is not known.
OBJECTIVE
We determined the ability of CYP2C19 and CYP3A4 to 21-hydroxylate progesterone and 17-hydroxyprogesterone (<em>17OHP</em>), determined the impact of the common P450 oxidoreductase (POR) variant A503V on these activities, and examined correlations between CYP2C19 variants and phenotype in patients with 21OHD.
METHODS
Bacterially expressed, N-terminally modified, C-His-tagged human P450c21, CYP2C19, and CYP3A4 were combined with bacterially expressed wild-type and A503V POR. The 21-hydroxylation of radiolabeled progesterone and <em>17OHP</em> was assessed, and the Michaelis constant (Km) and maximum velocity (Vmax) of the reactions were measured. CYP2C19 was genotyped in 21OHD patients with genotypes predicting severe congenital adrenal hyperplasia.
RESULTS
Compared to P450c21, the Vmax/Km for 21-hydroxylation of progesterone by CYP2C19 and CYP3A4 were 17 and 10%, respectively. With both forms of POR, the Km for P450c21 was approximately 2.6 microm, the Km for CYP2C19 was approximately 11 microm, and the Km for CYP3A4 was approximately 110 microm. Neither CYP2C19 nor CYP3A4 could 21-hydroxylate <em>17OHP</em>. The CYP2C19 ultrametabolizer allele CYP2C19 17 was homozygous in one of five patients with a 21OHD phenotype that was milder than predicted by the CYP21A2 genotype.
CONCLUSIONS
CYP2C19 and CYP3A4 can 21-hydroxylate progesterone but not <em>17OHP</em>, possibly ameliorating mineralocorticoid deficiency, but not glucocorticoid deficiency. Multiple enzymes probably contribute to extraadrenal 21-hydroxylation.
Related with
Pulse
Views:
1
Posts:
No posts
Not rated
Journal: Pediatrics
January/29/2002
Abstract
OBJECTIVE
To evaluate whether congenital adrenal hyperplasia (CAH) patients can be detected by newborn screening before the occurrence of life-threatening salt wasting and whether the prevalence, specificity, and sensitivity are adequate enough for a routine screening procedure.
METHODS
From 1998, a 2-year regional pilot screening for CAH was performed. In 1998, cutoff levels for <em>17OHP</em> were primarily based on birth weight, and in 1999 on gestational age. In addition, nationwide, all newly diagnosed patients with CAH were reported to the Dutch Pediatric Surveillance Unit to compare screened CAH patients with CAH patients in the area without screening.
RESULTS
In 2 years, 176 684 newborns were screened and 15 CAH patients (7 males/8 females) were detected. Therapy was started at the median age of 7 days. In the area without screening, 223 307 infants were born and 19 CAH patients (10 males/9 females) were reported to the Dutch Pediatric Surveillance Unit. Therapy was started at the median age of 14 days. The mean (standard deviation) serum sodium concentration was 134.5 (3.4) mmol/L in the area of screening versus 124.5 (10.8) mmol/L in the area without screening. The overall prevalence was 1:11 764. In 1998 and 1999, the specificity was 99.76% and 99.97%, respectively. The positive predictive value was 4.5% and 16%, respectively. To date, no false-negative cases have been detected.
CONCLUSIONS
Severe salt wasting can be prevented by neonatal screening. The prevalence, specificity, and sensitivity allowed addition of screening for CAH to the routinely performed national neonatal screening program.
Journal: JAMA - Journal of the American Medical Association
July/6/2011
Abstract
BACKGROUND
Lopinavir-ritonavir is a human immunodeficiency virus 1 (HIV-1) protease inhibitor boosted by ritonavir, a cytochrome p450 inhibitor. A warning about its tolerance in premature newborns was recently released, and transient elevation of 17-hydroxyprogesterone (<em>17OHP</em>) was noted in 2 newborns treated with lopinavir-ritonavir in France.
OBJECTIVE
To evaluate adrenal function in newborns postnatally treated with lopinavir-ritonavir.
METHODS
Retrospective cross-sectional analysis of the database from the national screening for congenital adrenal hyperplasia (CAH) and the French Perinatal Cohort. Comparison of HIV-1-uninfected newborns postnatally treated with lopinavir-ritonavir and controls treated with standard zidovudine.
METHODS
Plasma <em>17OHP</em> and dehydroepiandrosterone-sulfate (DHEA-S) concentrations during the first week of treatment. Clinical and biological symptoms compatible with adrenal deficiency.
RESULTS
Of 50 HIV-1-uninfected newborns who received lopinavir-ritonavir at birth for a median of 30 days (interquartile range [IQR], 25-33), 7 (14%) had elevated <em>17OHP</em> levels greater than 16.5 ng/mL for term infants (>23.1 ng/mL for preterm) on days 1 to 6 vs 0 of 108 controls having elevated levels. The median <em>17OHP</em> concentration for 42 term newborns treated with lopinavir-ritonavir was 9.9 ng/mL (IQR, 3.9-14.1 ng/mL) vs 3.7 ng/mL (IQR, 2.6-5.3 ng/mL) for 93 term controls (P < .001). The difference observed in median <em>17OHP</em> values between treated newborns and controls was higher in children also exposed in utero (11.5 ng/mL vs 3.7 ng/mL; P < .001) than not exposed in utero (6.9 ng/mL vs 3.3 ng/mL; P = .03). The median DHEA-S concentration among 18 term newborns treated with lopinavir-ritonavir was 9242 ng/mL (IQR, 1347-25,986 ng/mL) compared with 484 ng/mL (IQR, 218-1308 ng/mL) among 17 term controls (P < .001). The <em>17OHP</em> and DHEA-S concentrations were positively correlated (r = 0.53; P = .001). All term newborns treated with lopinavir-ritonavir were asymptomatic, although 3 premature newborns experienced life-threatening symptoms compatible with adrenal insufficiency, including hyponatremia and hyperkalemia with, in 1 case, cardiogenic shock. All symptoms resolved following completion of the lopinavir-ritonavir treatment.
CONCLUSIONS
Among newborn children of HIV-1-infected mothers exposed in utero to lopinavir-ritonavir, postnatal treatment with a lopinavir-ritonavir-based regimen, compared with a zidovudine-based regimen, was associated with transient adrenal dysfunction.
Related with
Pulse
Views:
1
Posts:
No posts
Not rated
Journal: Gynecological Endocrinology
August/29/2013
Abstract
OBJECTIVE
To evaluate the effects the administration of myo-inositol (MYO) on hormonal parameters in a group of polycystic ovary syndrome (PCOS) patients.
METHODS
Controlled clinical study.
METHODS
PCOS patients in a clinical research environment.
METHODS
50 overweight PCOS patients were enrolled after informed consent.
METHODS
All patients underwent hormonal evaluations and an oral glucose tolerance test (OGTT) before and after 12 weeks of therapy (Group A (n¼10): MYO 2 g plus folic acid 200 mg every day; Group B (n¼10): folic acid 200 mg every day). Ultrasound examinations and Ferriman-Gallwey score were also performed.
METHODS
Plasma LH, FSH, PRL, E2, <em>17OHP</em>, A, T, glucose, insulin, C peptide concentrations, BMI, HOMA index and glucose-to-insulin ratio.
RESULTS
After 12 weeks of MYO administration plasma LH, PRL, T, insulin levels and LH/FSH resulted significantly reduced. Insulin sensitivity, expressed as glucose-to-insulin ratio and HOMA index resulted significantly improved after 12 weeks of treatment. Menstrual cyclicity was restored in all amenorrheic and oligomenorrheic subjects. No changes occurred in the patients treated with folic acid.
CONCLUSIONS
MYO administration improves reproductive axis functioning in PCOS patients reducing the hyperinsulinemic state that affects LH secretion.
Journal: Steroids
March/15/2007
Abstract
Saliva analysis is an accepted non-invasive alternative to plasma in pediatric endocrinology. Although commercial saliva collectors are available, the reliability of these devices for the analysis of salivary hormones has not been proved. We investigated the recovery and linearity of salivary steroids (cortisol, cortisone, 17-hyroxyprogesterone, testosterone, androstenedione) being relevant in endocrine research and therapy control. Pooled saliva was spiked with ascending concentrations of the steroids and applied onto a variety of absorbents, such as the cotton and the polyester (PE) Salivette (Sarstedt), the foam-tip applicator (Whatman) and strips of blood-spot collection paper (Whatman). Analysis was performed by LC-MS/MS. Best results were achieved using the PE Salivette, yielding recoveries (%) of 99.8 (cortisol), 98.7 (cortisone), 91.8 (<em>17OHP</em>), 96.3 (testosterone), 98.9 (androstendione) with a volume recovery of 98+/-1%. Using the blood-spot paper, recoveries (%) were 92.0 (cortisol), 89.1 (cortisone), 72.0 (<em>17OHP</em>), 70.3 (testosterone) and 77.1 (androstendione). The recovery of glucocorticoids was significantly higher compared to androgens (p<0.001). The recovery of liquid volume was 95+/-2%. The cotton Salivette yielded weak recoveries of 88.7 (cortisol), 86.2 (cortisone), 60.9 (<em>17OHP</em>), 62.0 (testosterone) and 72.4 (androstendione). The recovery of the glucocorticoids differed significantly from the androgens (p<0.001). Liquid recovery was most variable with 89+/-8%. The weakest recoveries were found in the foam-tips being 76.2 for cortisol, only 41.8 for cortisone, 31.1 for <em>17OHP</em>, 38.5 for testosterone and 36.1 for androstendione. The volume recovery here was 97+/-1%. We assume only the PE version of the Salivette suitable for salivary steroid analysis. The weak recovery from the cotton version is a severe problem due to lacking comparability with values obtained with the polyester wads and the weak homogeneity as observed over a physiological concentration range.
Journal: Human Reproduction
March/24/2012
Abstract
BACKGROUND
Polycystic ovary syndrome (PCOS) patients typically have 17-hydroxyprogesterone (<em>17OHP</em>) hyperresponsiveness to GnRH agonist (GnRHa) (PCOS-T). The objective of this study was to determine the source of androgen excess in the one-third of PCOS patients who atypically lack this type of ovarian dysfunction (PCOS-A).
METHODS
Aged-matched PCOS-T (n= 40), PCOS-A (n= 20) and controls (n= 39) were studied prospectively in a General Clinical Research Center. Short (4 h) and long (4-7 day) dexamethasone androgen-suppression tests (SDAST and LDAST, respectively) were compared in subsets of subjects. Responses to SDAST and low-dose adrenocorticotropic hormone (ACTH) were then evaluated in all.
RESULTS
Testosterone post-SDAST correlated significantly with testosterone post-LDAST and <em>17OHP</em> post-GnRHa (r = 0.671-0.672), indicating that all detect related aspects of ovarian dysfunction. An elevated dehydroepiandrosterone peak in response to ACTH, which defined functional adrenal hyperandrogenism, was similarly prevalent in PCOS-T (27.5%) and PCOS-A (30%) and correlated significantly with baseline dehydroepiandrosterone sulfate (DHEAS) (r = 0.708). Functional ovarian hyperandrogenism was detected by subnormal testosterone suppression by SDAST in most (92.5%) PCOS-T, but significantly fewer PCOS-A (60%, P< 0.01). Glucose intolerance was absent in PCOS-A, but present in 30% of PCOS-T (P < 0.001). Most of the PCOS-A cases with normal testosterone suppression in response to SDAST (5/8) lacked evidence of adrenal hyperandrogenism and were obese.
CONCLUSIONS
Functional ovarian hyperandrogenism was not demonstrable by SDAST in 40% of PCOS-A. Most of these cases had no evidence of adrenal hyperandrogenism. Obesity may account for most hyperandrogenemic anovulation that lacks a glandular source of excess androgen, and the SDAST seems useful in making this distinction.
Journal: Pediatric Research
December/10/2000
Abstract
Some very preterm neonates admitted to the neonatal intensive care unit show circulatory and respiratory problems that improve after administration of steroids. It is unclear whether these symptoms could be caused by adrenal insufficiency. The objective of our study was to investigate the cortisol levels and the cortisol release from the adrenals after ACTH in very preterm infants with and without severe illness and to find whether a relation exists between adrenal function and outcome. An ACTH test (0.5 microg) was performed on d 4 in 21 very preterm infants (gestational age, 25.6-29.6 wk; birth weight, 485-1265 g). Baseline cortisol and 17-hydroxyprogesterone (<em>17OHP</em>) levels and the cortisol levels 30, 60, and 120 min after ACTH administration were measured. The Score for Neonatal Acute Physiology was used to measure illness severity. All infants showed an increase in cortisol levels after ACTH, but the cortisol levels were significantly lower in the ventilated more severely ill infants. After adjusting for birth weight and gestational age, the mean baseline cortisol levels and cortisol/<em>17OHP</em> ratios were significantly lower and the <em>17OHP</em> levels significantly higher in the ventilated infants compared with the nonventilated infants. Patients with an adverse outcome had significantly lower baseline cortisol/<em>17OHP</em> ratios and 60-min cortisol levels during ACTH testing (p = 0.002 and p = 0.03, respectively). These data suggest an insufficient adrenal response to stress in sick ventilated very preterm infants with gestational ages younger than 30 wk compared with nonventilated less sick preterm infants. Further studies are required to investigate whether supplementation with physiologic doses of hydrocortisone may benefit the outcome.
Journal: Clinical Chemistry
August/28/2006
Abstract
BACKGROUND
Congenital adrenal hyperplasia is a group of autosomal recessive disorders caused by a deficiency of 1 of 4 enzymes required for the synthesis of glucocorticoids, mineralocorticoids, and sex hormones. Analysis of 11-deoxycortisol (11DC), 17-hydroxyprogesterone (<em>17OHP</em>), 17-hydroxypregnenolone (<em>17OHP</em>r), and pregnenolone (Pr) in blood allows detection of these enzyme defects.
METHODS
The steroids were extracted from 200 microL of serum or plasma by solid-phase extraction, derivatized to form oximes, and extracted again with methyl t-butyl ether. Instrumental analysis was performed on an API 4000 tandem mass spectrometer with electrospray ionization in positive mode and multiple reaction-monitoring acquisition.
RESULTS
The limits of detection were 0.025 microg/L for 11DC, <em>17OHP</em>, and Pr and 0.10 microg/L for <em>17OHP</em>r. The method was linear to 100 microg/L for 11DC, <em>17OHP</em>, and Pr, respectively, and to 40 microg/L for <em>17OHP</em>r. Within- and between-run (total) imprecision (CVs) were <7.1% and 11%, respectively. Reference intervals for children in Tanner stages 1 through 5 and adult males and females for <em>17OHP</em>, 11DC, Pr, and <em>17OHP</em>r were established. Prepared samples were stable for >72 h.
CONCLUSIONS
The detection limit and selectivity of this method and its small sample volume requirement allow analysis of endogenous concentrations of adrenal steroids in serum or plasma from children and adults. The method thus has an important potential role in the evaluation of the status of 4 of the enzymes involved in adrenal steroid biosynthesis.
Journal: Journal of Clinical Endocrinology and Metabolism
July/2/1997
Abstract
Polycystic ovary syndrome (PCOS) is associated with chronic anovulation, hyperandrogenemia, insulin resistance (IR)/hyperinsulinemia, and a high incidence of obesity. Thus, PCOS serves as a useful model to assess the role of IR and chronic endogenous insulin excess on leptin levels. Thirty-three PCOS and 32 normally cycling (NC) women of similar body mass index (BMI) were studied. Insulin sensitivity (S(I)) was assessed by rapid ivGTT in a subset of 28 PCOS and 29 NC subjects; percent body fat was determined by dual-energy x-ray absorptiometry (DEXA) in 14 PCOS and 17 NC. Fasting (0800 h) and 24-h mean hourly insulin levels were 2-fold higher (P < 0.0001), and S(I) was 50% lower (P = 0.005) in PCOS than in NC, while serum androstenedione (A), testosterone (T), 17-alpha hydroxyprogesterone (<em>17OHP</em>), and estrone (E1) levels were elevated (P < 0.0001), and sex hormone-binding globulin (SHBG) levels were decreased (P < 0.01). Twenty-four hour LH pulse frequency, mean pulse amplitude, and mean LH levels were elevated in PCOS (P < 0.001) as compared with NC. Serum leptin levels for PCOS (24.1 +/- 2.6 ng/mL) did not differ from NC (21.5 +/- 3.5 ng/mL) and were positively correlated with BMI (r = 0.81) and percent body fat (r = 0.91) for the two groups (both P < 0.0001). Leptin levels for PCOS and NC correlated positively with fasting and 24-h mean insulin levels (r = 0.81, P < 0.0001 for both PCOS and NC) and negatively with S(I) and SHBG levels. Leptin concentrations for PCOS, but not NC, correlated positively with 24-h mean glucose levels and inversely with 24-h mean LH levels and 24-h mean LH pulse amplitude. Leptin levels were not correlated with estrogen or androgen levels for either PCOS or NC, although leptin levels were positively related to the ratios of E1/SHBG and E2/SHBG for both PCOS and NC and to the ratio of T/SHBG for PCOS only. In stepwise multivariate regression with forward selection, only 24-h mean insulin levels contributed significantly (P < 0.01) to leptin levels independent of BMI and percent body fat for both PCOS and NC. Given this relationship and the presence of 2-fold higher 24-h mean insulin levels in PCOS, the expected elevation of leptin levels in PCOS was not found. This paradox may be explained by the presence of adipocyte IR specific to PCOS, which may negate the stimulatory impact of hyperinsulinemia on leptin secretion, a proposition requiring further study.
Journal: Journal of Clinical Endocrinology and Metabolism
October/15/2000
Abstract
Bone morphogenetic proteins (BMPs), members of the transforming growth factor beta superfamily, were recently shown to be expressed and to regulate steroidogenesis in rat ovarian tissue. The purpose of this study was to investigate the effect of BMP-4 on androgen production in a human ovarian theca-like tumor (HOTT) cell culture model. We have previously demonstrated the usefulness of these cells as a model for human thecal cells. HOTT cells respond to protein kinase A agonists by increased production of androstenedione and with an induction of steroid-metabolizing enzymes. In this investigation, HOTT cells were treated with forskolin or dibutyryl cyclic AMP (dbcAMP) in the presence or absence of various concentrations of BMP-4. The accumulation of androstenedione, progesterone, and 17alpha-hydroxyprogesterone (<em>17OHP</em>) in the incubation medium was measured by RIA. The expression of 17alpha-hydroxylase (CYP17), 3beta-hydroxysteroid dehydrogenase (3betaHSD), cholesterol side-chain cleavage (CYP11A1), and steroidogenic acute regulatory (StAR) protein was determined by protein immunoblotting analysis using specific rabbit polyclonal antibodies. We also examined the expression of BMP receptor subtypes in our HOTT cells using RT-PCR. In cells treated with medium alone, steroid accumulation and steroid enzyme expression was unchanged. In cells treated with BMP alone there was a modest decrease in androstenedione secretion. In the presence of forskolin, HOTT cell production of androstenedione, <em>17OHP</em>, and progesterone increased by approximately 4.5-, 35-, and 3-fold, respectively. In contrast, BMP-4 decreased forskolin-stimulated HOTT cell secretion of androstenedione and <em>17OHP</em> by 50% but increased progesterone production 3-fold above forskolin treatment alone. Forskolin treatment led to an increase in CYP17, CYP11A1, 3betaHSD, and StAR protein expression. BMP-4 markedly inhibited forskolin stimulation of CYP17 expression but had little effect on 3betaHSD, CYP11A1, or StAR protein levels. Similar results were observed with the cAMP analog dbcAMP. In addition, BMP-4 inhibited basal and forskolin stimulation of CYP17 messenger RNA expression as determined by RNase protection assay. Other members of the transforming growth factor beta superfamily, including activin and inhibin, had minimal effect on androstenedione production in the absence of forskolin. In the presence of forskolin, activin inhibited androstenedione production by 80%. Activin also inhibited forskolin induction of CYP17 protein expression as determined by Western analysis. We identified the presence of messenger RNA for three BMP receptors (BMP-IA, BMP-IB, and BMP-II) in the HOTT cells model. In conclusion, BMP-4 inhibits HOTT cell expression of CYP17, leading to an alteration of steroidogenic pathway resulting in reduced androstenedione accumulation and increased progesterone production. These effects of BMP-4 seem similar to those caused by activin, another member of the transforming growth factor-beta superfamily of proteins.
Journal: Clinical Endocrinology
December/13/2015
Abstract
BACKGROUND
Nonclassical congenital adrenal hyperplasia (NC-CAH) is caused by mutations of the CYP21A2 gene. The clinical manifestations and hormonal derangements of NC-CAH are quite variable.
OBJECTIVE
(i) To define the phenotype and its relation to genotype according to gender and age and (ii) to evaluate the validity of currently applied hormonal criteria for establishing the diagnosis of NC-CAH.
METHODS
The clinical, hormonal and molecular data of 280 subjects (235 female) with NC-CAH and a median age of 17·6 years were analysed. CYP21A2 genotyping was performed in all subjects.
RESULTS
The majority of females aged less than 8 years presented with premature pubarche (88·3%), while those older than 8 presented with a polycystic ovary-like phenotype (63·2%). A total of 7·7% of the females and 51·1% of the males were asymptomatic at the time of diagnosis. In the total group, 50·4% of the subjects were compound heterozygotes for one classical (C) and one nonclassical (NC) mutation, while 46% of the alleles studied carried the p.V281L mutation. Basal <em>17OHP</em> values were below 6 nm (2 ng/ml) in 2·1% of the subjects with NC-CAH, but none had peak <em>17OHP</em> values post-ACTH lower than 30 nm (10 ng/ml).
CONCLUSIONS
NC-CAH has a variable phenotype depending on the age, gender and the presence of a classical mutation. A peak cut-off value of <em>17OHP</em> post-ACTH lower than 30 nm excludes the diagnosis of NC-CAH, whereas basal <em>17OHP</em> <6 nm may represent a false-negative result. A significant number of patients harboured a classical mutation, a finding which requires genotyping of the partner for genetic counselling.
Journal: American Journal of Obstetrics and Gynecology
January/15/1981
Abstract
This study was carried out to document the postpubertal presentation of congenital adrenal hyperplasia (CAH), to elaborate the diagnostic criteria for it, and to investigate family members of CAH patients. Serum 17-hydroxyprogesterone (<em>17OHP</em>) was measured in normal women and 25 hirsute oligomenorrheic patients, five of whom were shown to have CAH. These five CAH patients, as a group, had significantly elevated levels of <em>17OHP</em> when compared to normal and hirsute women, although the other 20 hirsute oligomenorrheic women also had higher levels of <em>17OHP</em> than the follicular phase control subjects. A single intravenous bolus of 0.25 mg of adrenocorticotropic hormone (ACTH) caused much larger increased in <em>17OHP</em> in all five CAH patients than in the control and hirsute women. The five CAH patients had decreased cortisol but normal 11-deoxycortisol responses to ACTH, thus indicating 21-hydroxylase deficiency (21HD). Clinically, they were indistinguishable from women with polycystic ovarian disease (PCO) and had basal serum levels of androgens and urinary 17-ketosteroids which were similar to those found in 47 other women presenting with the complaint of hirsutism. However, the androstenedione levels and androstenedione/cortisol ratios in response to ACTH were significantly higher in the five CAH patients than in both the normal and hirsute women. Of seven family members tested, two fathers and one mother had an intermediate <em>17OHP</em> response to ACTH, thus suggesting heterozygosity. Human lymphocyte antigen (HLA) typing on family members indicated that the inheritance of the disorder may be linked to B antigens. Two siblings of one of the CAH patients had normal <em>17OHP</em> responses to ACTH and also had a different HLA-B complement. These data document the existence of adult manifestation of CAH, due to 21 HD. This disorder presents with androgen excess and oligomenorrhea or amenorrhea and mimicks PCO. The diagnosis of it hinges upon the post-ACTH rise in <em>17OHP</em>, whereas the levels of serum androgens and urinary 17-ketosteroids may be inconclusive.
Search and select entities to see details here. Details will include lists of related entities (via publication). Explore related entities further.