The suggestion that androgens may regulate testosterone (T) production in rat Leydig cells by a receptor-mediated feed-back mechanism, led us to investigate whether in vivo the absence of testicular androgen receptors, as it occurs in testicular feminization (TF), may modify the characteristic testicular response observed in men and prepubertal children after a single dose of hCG. Subjects consist of: 1) six normal men, 2) two adult patients with the complete form of androgen insensitivity syndrome (TF), 3) 12 normal prepubertal boys, 4) one prepubertal boy with the same form of TF. Each subject received i.m. a single dose of hCG 3500 IU/m2 b.s. and blood samples were collected basally and 2, 4, 24, 48, 72 and 96 hours after the hormonal stimulus. Serum levels of T, 17 alpha hydroxyprogesterone (<em>17OHP</em>) and 17 beta estradiol (E2) were measured at each collection time. In normal men a significant increase in T (M +/- SE) was observed at 4 h (758.6 +/- 135 ng/dl, P less than 0.05) and a more significant increase at 48 h (1082 +/- 60.3 ng/dl, P less than 0.001). E2 and <em>17OHP</em> peaked significantly at 24 h (81.5 +/- 9.6 pg/ml and 460.7 +/- 90.9 ng/dl respectively). This response pattern is characteristic of the testicular desensitization which occurs in normal man after a single hCG dose. The same response pattern has been observed in the two TF adult patients suggesting that human testicular desensitization in vivo does not depend on androgen receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Objectives Our recent survey of 44 mass spectrometry laboratories across 17 countries identified variation in internal standard (IS) choice for the measurement of serum/plasma 17α-hydroxyprogesterone (<em>17OHP</em>) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The choice of IS may contribute to inter-method variations. This study evaluated the effect of two common isotopically labeled IS on the quantification of <em>17OHP</em> by LC-MS/MS. Methods Three collaborating LC-MS/MS laboratories from Asia, Europe and Australia, who routinely measure serum <em>17OHP</em>, compared two IS, (1) IsoSciences carbon-13 labeled <em>17OHP</em>-[2,3,4-13C3], and (2) IsoSciences deuterated <em>17OHP</em>-[2,2,4,6,6,21,21,21-2H]. This was performed as part of their routine patient runs using their respective laboratory standard operating procedure. Results The three laboratories measured 99, 89, 95 independent samples, respectively (up to 100 nmol/L) using the 13C- and 2H-labeled IS. The slopes of the Passing-Bablok regression ranged 0.98-1.00 (all 95% confidence interval [CI] estimates included the line of identity), and intercept of <0.1 nmol/L. Average percentage differences of -0.04% to -5.4% were observed between the two IS materials, which were less than the optimal bias specification of 7% determined by biological variation, indicating no clinically significant difference. The results of 12 Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP) proficiency samples (1-40 nmol/L) measured by the laboratories were all within the RCPAQAP analytical performance specifications for both IS. Conclusions Overall, the comparison between the results of 13C- and 2H-labeled IS for <em>17OHP</em> showed good agreement, and show no clinically significant bias when incorporated into the LC-MS/MS methods employed in the collaborating laboratories.

Keywords: 17α-hydroxyprogesterone; internal standard; mass spectrometry.

Immunochemical measurement of serum 17α-hydroxyprogesterone (<em>17OHP</em>), the most important parameter for diagnosis of classical 21-hydroxylase deficiency (21OHD) in newborn infants, is known to be inaccurate due to the cross-reactivity of antibodies with a large quantity of fetal adrenal steroids. The aims of this study were 1) to establish reference values for the serum <em>17OHP</em> level in Japanese newborn infants using non-immunochemical stable isotope dilution -gas chromatography/mass spectrometry (SID-GC/MS) and 2) to compare the serum <em>17OHP</em> levels determined by SID-GC/MS with those determined by radioimmunoassay (RIA). The first study subjects were used for determination of reference values and included 57 healthy full-term newborn infants (4-5 d of age). The second study subjects were used for comparison of SID-GC/MS with RIA and included 27 healthy full-term newborn infants (3-6 d of age) and two subjects with neonatal transient hyper <em>17OHP</em>nemia; these two subjects were 16 and 27 d of age, respectively. In the first study subjects, the intra-assay coefficient of variation for SID-GC/MS was 3% (n=5), the recovery rate was 98%, the sensitivity was 0.2 ng/ml, and the range of linearity was 0.5-200 ng/ml. The reference values for the serum <em>17OHP</em> level determined by SID-GC/MS ranged from 0.3-1.5 (0.6) (ng/ml) (median). In the second study subjects, the serum <em>17OHP</em> levels determined by SID-GC/MS were lower in one of the 27 subjects and both of the two subjects with neonatal transient hyper <em>17OHP</em>nemia compared with the levels determined by RIA. Measurement of the serum <em>17OHP</em> level using SID-GC/MS may be clinically useful for definitive diagnosis of classical 21OHD in newborn infants.

The measurements of steroids in biological fluids are of importance for the diagnosis and treatment of many diseases. Liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) has a high specificity and accuracy for the steroid analysis, whereas it has a lower analysis throughput, which could become a big issue in clinical practice. One of the promising solutions to this issue is the multiplexing of samples in the same injection. In this study, the utility of the sample-multiplexing by the derivatization using multiple analogous reagents was evaluated for enhancing the throughput of the LC/ESI-MS/MS assays of steroids. The plasma 17α-hydroxyprogesterone (<em>17OHP</em>), which is a diagnostic marker for the 21-hydroxylase deficiency, was chosen as the model analyte. The four plasma samples (20 μL each) were separately derivatized with one of four different analogous Girard-type reagents, combined, then injected together into the LC/ESI-MS/MS. By this method, four plasma samples could be analyzed within a single LC run. The developed method could significantly reduce the total LC run time (about 2/5 for 32 samples, compared with the conventional method) with a satisfactory sensitivity (lower limit of quantification 0.5 ng/mL), precision (intra- and inter-assay RSDs ≤ 4.0% and ≤ 3.5%, respectively) and accuracy (97.6-106.7%), and negligible matrix effect. The developed method had a satisfactory applicability for the quantification of <em>17OHP</em> in the cord plasma samples.

Deficiency of 21-hydroxylase constitutes the most frequent form of Congenital adrenal hyperplasia (CAH) and is classified into classical and nonclassical (NC) forms. The type of the molecular defect determines the severity of the phenotype with a high degree of concordance for the classical genotypes but with lower concordance for the NC genotypes. The clinical expression of NC-CAH is quite variable ranging from an asymptomatic state to premature pubarche, hirsutism, acne, menstrual irregularities, infertility and increased rate of spontaneous abortions. Basal and/or ACTH stimulated 17-hydroxyprogesterone (<em>17OHP</em>) values,>> 5 ng/ml and >10 ng/ml, respectively are currently applied to diagnose NC-CAH. Glucocorticoid administration to patients with NC-CAH is determined by the clinical manifestations. During pregnancy, steroid replacement therapy and adequate monitoring reduce the risk of spontaneous abortions. CYP21A2 genotyping of the partner of a prospective mother with NC-CAH is controversial. Most likely it should be performed if the mother carries a classical mutation.

Late-onset congenital adrenal hyperplasia is a cause of a spectrum of clinical manifestations of postnatal androgen excess. In these cases, ACTH stimulation test with measurement of 17-Hydroxyprogesterone (<em>17OHP</em>) is usually done to assess 21-hydroxylase (21-OH) deficiency. Determination of the 11-deoxycortisol (S) and the S to cortisol ratio is rarely done, so that 11 beta-hydroxylase (11-OH) deficiency seems unusual. We systematically investigated this biosynthetic defect among women complaining of hyperandrogenism (n = 519) and, comparing the patient's hormonal responses to ACTH with those of 31 normal women, found 29 11-OH deficiency (5.6%): this is the largest group ever reported. S was elevated only 9 times, so that using this single determination, diagnosis of 20 enzymatic defects would not have been made. Only three of the patients (10%) had hypertension, even though the pathway of aldosterone was involved in 33% of cases (criteria: elevation of the ratio desoxycorticosterone to corticosterone). We also described one new patient with both 11-OH and 3-beta-hydroxysteroid dehydrogenase deficiencies. The patho-physiology is particularly interesting in these cases. It is concluded that the single research for 21-OH deficiency is inadequate among women complaining of hyperandrogenism: the screening for 11-OH deficiency should be made, even if blood pressure is normal.

Introduction. Adrenal glands play a major role in the control of blood pressure and mild defects of steroidogenesis and/or inappropriate control of mineralocorticoid production have been reported in high blood pressure (HBP). Patients and Methods. We used a specific protocol for the evaluation of 100 consecutive patients with inappropriate or recent onset HBP. Specific methods were used to confirm HBP and to diagnose secondary forms of HBP. In addition we tested adrenal steroidogenesis with the common cosyntropin test, modified to include the simultaneous measurement of renin and aldosterone besides 17-hydroxyprogesterone (<em>17OHP</em>) and 11-deoxycortisol (S). Results. Secondary forms of HBP were diagnosed in 32 patients, including 14 patients with primary hyperaldosteronism (PA) (14%) and 10 patients with pheochromocytoma (10%). Mild defects of the 21-hydroxylase (21OHD) and 11-hydroxylase (11OHD) enzymes were common (42%). ACTH-dependent aldosterone secretion was found in most patients (54%) and characteristically in those with mild defects of adrenal steroidogenesis (>60%), PA (>75%), and otherwise in patients with apparent essential HBP (EHBP) (32%). Discussion. Mild defects of adrenal steroidogenesis are common in patients with HBP, occurring in almost half of the patients. In those patients as well as in patients with apparent EHBP, aldosterone secretion is commonly dependent on ACTH.

This study was designed to investigate the effect of steroid administration in ill premature neonates. Twenty high-risk very-low-birth-weight (VLBW) infants [birth weight (BW) < or = 1,300 g] with a mean BW 948 +/- 220 g, gestational age (GA) 27 +/- 1.7 weeks underwent 1-hour ACTH (Cortrosyn) stimulation tests and determination of 17-hydroxyprogesterone (<em>17OHP</em>)/dehydroepiandrosterone sulfate (DHEAS) at 23.6 +/- 15.9 days poststeroid treatment for bronchopulmonary dysplasia (BPD)/airway obstruction. Metyrapone tests were also obtained in 18 infants. Baseline (nonsteroid-exposed) values for pre-/post-ACTH cortisol, <em>17OHP</em>, DHEAS, and pre-/post-metyrapone compound S values were obtained in 5 infants. Eight of 18 (44%) infants had evidence of secondary (hypothalamic-pituitary) adrenal suppression based on abnormal metyrapone tests. No difference was found in BW, GA, time on O2 or AV, steroid dose/kg, or neonatal/postneonatal mortality between the suppressed and nonsuppressed groups. Two of 4 infants with borderline ACTH tests had subnormal compound S levels postmetyrapone. No relationship was found between steroid dose/kg and cortisol response post-ACTH. Additionally, corrected GA was not related to change in cortisol, <em>17OHP</em>, and DHEAS pre-/post-ACTH. Two infants exhibited recovery of adrenal suppression documented by repeated metyrapone testing at 63 and 186 days poststeroid treatment. In conclusion, this study documents the apparent high incidence of secondary adrenal suppression in VLBW infants treated with dexamethasone. Clinical significance of these findings deserves further investigation.

UNASSIGNED

Hypercalcemia is reported as a rare finding in adrenal insufficiency, but is not well described in congenital adrenal hyperplasia (CAH).

UNASSIGNED

A retrospective chart review was conducted of patients with CAH diagnosed before the age of 2 years who had at least one recorded serum calcium measurement. Data from birth to 6 years of age were reviewed.

UNASSIGNED

Of the 40 patients who met inclusion criteria, 33 (82.5%) had at least one elevated calcium concentration and 21 (53%) had two or more elevated calcium concentrations. Of the 126 elevated serum calcium concentrations, the median was 10.9 mg/dL (range, 10.6 to 14.2 mg/dL). Median age at the last elevated calcium measurement was 5 months (range, 0.3 to 46 months). Serum calcium concentration was inversely related to age (r = -0.124; P = 0.004). Overall, calcium level positively correlated with 17-hydroxyprogesterone (<em>17OHP</em>) concentration (r = 0.170; P = 0.003), and this remained significant after adjusting for age (P < 0.05). However, patients had hypercalcemia with both high and low <em>17OHP</em> concentrations. Serum calcium concentration also was positively related to glucocorticoid (r = 0.196; P = 0.012) and fludrocortisone (r = 0.229; P = 0.003) doses, and remained significant after age adjustment. Only seven patients were evaluated for hypercalciuria. Of these, six had at least one period of documented hypercalciuria. Three patients had nephrocalcinosis on renal ultrasound.

UNASSIGNED

Children with CAH are at risk for developing hypercalcemia, hypercalciuria, and nephrocalcinosis. Further studies are needed to determine the broader prevalence and the etiology of hypercalcemia in CAH.

21-hydroxylase deficiency (21OHD) is a rare genetic disorder in which salt-wasting syndrome occurs in 75% of cases, due to inability to synthesize cortisol and aldosterone. Recent mass spectrometry progress allowed identification of 21-deoxysteroids, i.e 17-hydroxyprogesterone (<em>17OHP</em>), 21-deoxycortisol (21DF), 21-deoxycorticosterone (21DB). We hypothesized that they may interfere with mineralocorticoid signaling and fludrocortisone therapy in patients with congenital adrenal hyperplasia (CAH) without effective glucocorticoid replacement and ACTH suppression. Our goal was to quantify circulating 21-deoxysteroids in a pediatric cohort with CAH related to 21OHD and to examine their impact on mineralocorticoid receptor (MR) activation. Twenty-nine patients with salt-wasting phenotype were classified in two groups according to their therapeutic control. During routine follow-up, <em>17OHP</em>, 21DF, 21DB and cortisol levels were quantified by LC-MS/MS before hydrocortisone intake and 1 and 2.5 h following treatment administration. Luciferase reporter gene assays were performed on transfected HEK293T cells while <i>in silico</i> modelling examined structural interactions between these steroids within ligand-binding domain of MR. Plasma <em>17OHP</em>, 21DF and 21DB accumulate in uncontrolled patients reaching micromolar concentrations even after hydrocortisone intake. 21DF and 21DB act as partial MR agonists with antagonist features similar to <em>17OHP</em>, consistent with altered anchoring to Asn⁷⁷⁰ and unfavorable contact with Ala⁷⁷³ in ligand-binding pocket of MR. Our results demonstrate complex interaction between all accumulating 21-deoxysteroids in uncontrolled 21OHD patients and mineralocorticoid signaling and suggest that appropriate steroid profiling should optimize management and follow-up of such patients, as keeping those steroids to low plasma levels should attest therapeutic efficacy and prevent interference with MR signaling.

<AbstractText>Screening for and diagnosing non classic congenital adrenal hyperplasia (NCCAH) uses serum 17-hydroxyprogesterone (<em>17OHP</em>) thresholds established from immunoassay data; however, a new liquid-chromatography tandem mass spectrometry (LC-MS/MS) method results in lower <em>17OHP</em> values. The evolution of immunoassays is also challenging our diagnostic cut-off for glucocorticoid insufficiency and few data re-evaluate the utility of testing for glucocorticoid insufficiency in NCCAH.</AbstractText><AbstractText>(1) Evaluate the <em>17OHP</em> threshold that predicts NCCAH in children using LC-MS/MS, and (2) determine the prevalence of glucocorticoid insufficiency in NCCAH.</AbstractText><AbstractText>A retrospective chart review of pediatric patients who underwent ACTH stimulation tests with cortisol and <em>17OHP</em> measurements from 2011 to 2018 for assessment of NCCAH. Other adrenal pathologies were excluded. A cortisol < 415 nmol/L defined glucocorticoid insufficiency. Published correlation data determined a <em>17OHP</em> of 3.3 nmol/L by LC-MS/MS was equivalent to 6 nmol/L by immunoassay. Data analysis was by measures of diagnostic accuracy.</AbstractText><p><div><b>Results</b></div>Of 188 patients included, 23 (12%) had NCCAH (21/23 had genetic confirmation); the remaining 2 had peak <em>17OHP</em> <b>></b> 30 nmol/L. Baseline <em>17OHP</em> <b>≥</b> 6 nmol/L most accurately screened for NCCAH-sensitivity and specificity 96%. Almost all genetically confirmed NCCAH (20/21) had peak <em>17OHP</em> > 30 nmol/L; all subjects with other diagnoses peaked < 30 nmol/L. Glucocorticoid insufficiency was present in 55% with NCCAH.</p><p><div><b>Conclusions</b></div>Despite the increased specificity of LC-MS/MS, a baseline <em>17OHP</em> <b>≥</b> 6 nmol/L most accurately screened for NCCAH; this supports current practice guidelines. This threshold identified all with glucocorticoid insufficiency, notably prevalent in our cohort and for whom glucocorticoid stress dosing should be considered.</p>

<strong class="sub-title"> Background: </strong> Mutations of <i>CYP21A2</i> encoding 21-hydroxylase are the most frequent cause of congenital adrenal hyperplasia (CAH) and are associated either with elevated basal or ACTH-stimulated levels of 17-hydroxyprogesterone (<em>17OHP</em>) in blood.

Objective: The study objective was to identify the most suitable of 12 different test algorithms and appropriate cut-off levels for that test to recognize patients with non-classical congenital adrenal hyperplasia (NCCAH) and carriers of clinically relevant mutations in CYP21A2.

<strong class="sub-title"> Method and patients: </strong> Between July 2006 and July 2015 ACTH-tests were conducted in 365 children and adolescents (Age 1-20 y) suspected to have NCCAH. As a reference, results from subsequent gene sequencing of <i>CYP21A2</i> was used. Inclusion criteria that were used were premature pubarche with accelerated bone age, hyperandrogenism, hirsutism, or menstrual irregularities. Receiver operating characteristics (ROC) were plotted. Evaluated test algorithms were composed around <em>17OHP</em> measurements by radioimmunoassays. The most suitable test was identified by the greatest area under the curve (AUC).

<strong class="sub-title"> Results: </strong> Among the 12 tested algorithms, the sum of 30 min and 60 min stimulated <em>17OHP</em> values (sum<em>17OHP</em>stim) showed the highest AUC of 0.774 for identifying heterozygous and bi-allelic mutations. A cut-off of 10.1 μg/l was advisable. Bi-allelic mutations only were best identified calculating the difference between 30 min and basal <em>17OHP</em> values (Δ<em>17OHP</em>30). A cut-off of 9.4 μg/l was most effective.

<strong class="sub-title"> Conclusion: </strong> Alternatively to the above mentioned cut-offs the difference of 60 min after stimulation to basal <em>17OHP</em> (Δ<em>17OHP</em>60) can be used for the benefit of a combined test to identify both heterozygotes and bi-allelic patients. There are minimal decreases in sensitivity and specificity compared to an approach that applies two tests. However, it denotes a simpler approach in the clinical routine.

There are few reports of an association between Turner syndrome (TS) and 21-hydroxylase deficiency. However, this association is more frequent in some populations. The aim of this study was to evaluate the incidence of 21-hydroxylase deficiency in patients with TS in our population. 21-hydroxylase deficiency was evaluated in 44 TS cases with 45X (n=20) and 24 mosaic cases. A standard dose adrenocorticotropic (ACTH) stimulation test (Synacthen, Novartis, Basel, Switzerland) was performed, and 17 hydroxyprogesterone (<em>17OHP</em>), dehydroepiandrosterone sulfate (DHEAS) and cortisol responses were evaluated. Patients with increased <em>17OHP</em> responses in the stimulation test also underwent 21-hydroxylase gene analysis. The mean age was 14.6 +/- 4 (2.6-22.4); 37 patients were on growth hormone (GH) treatment. Nine patients were at prepubertal stage, whereas 35 were pubertal (24 on gonadal steroids and 11 spontaneously). Six patients were obese. Only one of our patients had a level of 7.5 ng/mL of <em>17OHP</em>, and there was no mutation found in congenital adrenal hyperplasia (CAH) genetic analysis. In other cases, peak <em>17OHP</em> levels were < or = 6 ng/mL. The mean peak <em>17OHP</em> was 2.62 +/- 1.48 (1.19-7.5) ng/mL, the cortisol level was 37.6 +/- 8.43 (23.9-56.2) microg/dL and the DHEAS was 135.2+/- 87.3 (15-413) microg/dL. The increased mean basal and peak cortisol levels (20.5 +/- 10.2 and 37.6 +/- 8.4 microg/dL) were remarkable findings. Whereas basal cortisol was above 20 microg/dL in 38.7% of patients, exaggerated results up to 56.2 microg/dL were obtained in peak cortisol levels. The basal and peak <em>17OHP</em> cortisol levels were not correlated with the presence of puberty, chromosome structure, gonadal steroid use, obesity or growth hormone use. This trial suggested that 21-hydroxylase deficiency was not common among patients with TS in our population. Adrenal function should be assessed, at least in the presence of clitoral enlargement in patients with TS, particularly if their karyotype does not contain a Y chromosome.

We report on a case of a 63-year-old male patient who presented with an adrenal incidentaloma corresponding to a pre-toxic adrenocortical adenoma ("pre-Cushing's syndrome"). Nycthemeral cortisol cycle and free urinary cortisol were within the normal range. Basal ACTH and dehydroepiandrosterone sulfate levels were decreased and the 17-hydroxyprogesterone (<em>17OHP</em>) response under ACTH stimulation (tetracosactide) was increased. The ACTh test appears mandatory in the incidentaloma work-up to order to identify the pre-toxic adenomas. Its elevation reflects likely intra-tumoral enzymatic defects. Pre-Cushing's syndrome therapeutic management is still debated. No surgery was performed in our patient. A 3-year follow-up did not show overt Cushing's syndrome features.

ACTH and 17-hydroxyprogesterone (<em>17OHP</em>) levels during clitoro- and labinoplasty for CYP21A2 deficiency have not been reported. In this study, we measured these levels during surgery. Intensive glucocorticoid supplement (IGS Tx; intravenous bolus injection of daily dose (14.6 to 22.2 mg/m(2)) of hydrocortisone followed by continuous infusion of three times the daily dose for 24 hours) was done to eight patients for surgery. ACTH and <em>17OHP</em> levels were generally suppressed during operation by this protocol, but mid-surgical elevations of ACTH and <em>17OHP</em> levels were observed in four out of the eight. In another three patients than the eight as described, oral dexamethasone pretreatment (1 mg/m(2)) was administered for two days before surgery in addition to IGS Tx. ACTH and <em>17OHP</em> levels were completely suppressed throughout operation, indicating that this kind of additional pretreatment is more effective approach.

<AbstractText> Weekly 17-hydroxyprogesterone caproate (<em>17OHP</em>-C) from 16 to 36 weeks' gestation reduces the risk of recurrent spontaneous preterm birth (sPTB). Limited data suggest poor adherence to published guidelines. This study aimed to identify factors associated with <em>17OHP</em>-C utilization.</AbstractText><AbstractText> This retrospective cohort study included women with a singleton pregnancy who delivered within one academic health system between January 2014 and December 2015. Eligible women had a history of ≥1 singleton sPTB. Primary outcomes were counseling about, receipt of, and adherence to <em>17OHP</em>-C therapy. Demographic and clinical predictors of the primary outcomes were determined using stepwise logistic regression.</AbstractText><p><div><b>RESULTS</b></div> Of 410 eligible subjects, 69% (<i>N</i> = 284) were counseled about and 36% (<i>N</i> = 148) received <em>17OHP</em>-C. Hispanic ethnicity, delay in prenatal care initiation, and a history of prior term births were associated with lower odds of <em>17OHP</em>-C counseling. Each week delay in prenatal care initiation, Hispanic ethnicity, and each additional week's gestation of the earliest prior sPTB decreased the odds of receiving <em>17OHP</em>-C. Maternal age and prior term births were associated with adherence.</p><AbstractText> Utilization of evidence-based <em>17OHP</em>-C therapy remains suboptimal: cultural and access-to-care barriers for eligible women may impede efforts to decrease recurrent sPTB rates.</AbstractText>

Objective The objective of this study was to determine if maternal smoking modifies the effectiveness of 17 α-hydroxyprogesterone caproate (<em>17OHP</em>-C). Study Design Secondary analysis of the Maternal-Fetal Medicine Units Network trial of <em>17OHP</em>-C. The prevalence of preterm birth (PTB) by smoking status and treatment group was compared by chi-squared analysis and analysis of variance was used to compare gestational age (GA) at birth. Multivariable modeling was used to estimate the effect of smoking on <em>17OHP</em>-C treatment. Results In this study, 459 women were included. Maternal smoking significantly modified the effectiveness of <em>17OHP</em>-C treatment. In smokers, <em>17OHP</em>-C significantly reduced the prevalence of multiple outcomes (PTB < 37 and < 35 weeks, spontaneous PTB < 37 and < 35 weeks), while in nonsmokers, only PTB < 37 weeks was reduced. Delivery GA was later in <em>17OHP</em>-C versus placebo treated smokers (36.4 vs. 34.3 weeks, p = 0.041) but not nonsmokers (36.3 vs. 35.5 weeks, p = nonsignificant). In multivariable modeling, <em>17OHP</em>-C was more effective in smokers than nonsmokers as measured by multiple outcomes (PTB < 37 weeks [p = 0.041] and < 35 weeks [p = 0.036] and spontaneous PTB < 37 weeks [p = 0.029]). Conclusion In this cohort of women with a prior PTB, maternal smoking status significantly modified the effectiveness of <em>17OHP</em>-C treatment.

Basal serum concentrations of cortisol, dehydroepiandrosterone (DHA) and its sulfate (DHAS), 4-androstene-3,17-dione (A4) and 17 alpha-hydroxyprogesterone (<em>17OHP</em>) were measured yearly in children treated with bone marrow transplantation (BMT) with or without preceding total body irradiation (TBI). Age-matched controls were used for comparison. ACTH stimulation tests were performed in the patients before and after treatment. However, in the samples taken before BMT only cortisol was measured. Basal posttreatment cortisol levels were subnormal in TBI-treated boys (n = 14, aged 5-17 years at BMT) during the adrenarcheal period (7-14 years) but became normal afterwards. All other groups had normal cortisol values. Treatment neither affected basal levels nor the ACTH-induced increment (delta-value) of cortisol. In the boys treated with TBI, normal basal levels of <em>17OHP</em> and adrenal androgens were found with the exception of decreased DHA levels in the postadrenarcheal boys. However, the delta-<em>17OHP</em> values and had an abnormal age relation and were significantly higher than in the patients not treated with TBI. In the patients not treated with TBI (6 boys aged 2-17 years) normal responses were found for 5 years or more after treatment. In female patients treated with TBI (n = 12, aged 1-16 years) circulating levels of DHA, DHAS and A4 were significantly decreased up to 5 years or more following treatment. It is concluded that after TBI, the cortisol homeostasis is maintained at the cost of reduced adrenal androgen secretion.

Changes in levels of sex steroids and gonadotropins were measured in 16 normal prepubertal and 15 pubertal girls prior to and after a 3 hour infusion of 100 microgrm synthetic gonadotropin releasing hormone (Gn-RH). Plasa estradiol (E2) concentrations rose significantly (p less than 0.02) from 29.7 +/- 4.6 (SE) pg/ml in the basal period to to 46.8 +/- 7.1 at the end of the infusion in the pubertal girls but were unchanged in the prepubertal girls. Estrone (E1), progesterone (P), 17-HYDROXYPROGESTERONE (<em>17OHP</em>), TESTOSTERONE (T), DIHYDROTESTOSTERONE (DHT), and androstenedione (A), dehydroepiandrosterone (DHA) and dehydroepiandrosterone sulfate (DHAS) levels were not altered in either maturity group. Basal plasma E2, E1, T, DHT, DHA and DHAS concentrations significantly correlated with the releasable pool of LH evoked by Gn-RH from the pituitary gonadotropes. We conclude: 1) The ovary is not highly and rapidly responsive to transient elevations of endogenous gonadotropin, and 2) Adrenal androgens may to some extent modulate the maturation of the hypothalamic-pituitary-gonadal system, at least as reflected by the pituitary response to exogenous Gn-RH.

The aim of our study was to estimate the adrenal function in hirsute girls.

METHODS

57 girls with hirsutism aged from 12 to 19 years, mean age 15.95 years, were involved into the study. The research was performed in early and middle follicular stage. Menstrual disorders were observed in 78% of them. Hirsutism was estimated with Ferriman-Gallwey scale (mean value 13+/-1.58), mean BMI was 22.7. The patients were divided into 3 groups: group 1 with clinical and laboratory symptoms of PCOS, n=29; group 2 with menstrual disorders and without elevated androgen level, n=15; group 3 without menstrual disorders and without elevated androgen level, n=13.

RESULTS

<em>17OHP</em> level was the highest in group I (1.17+/-0.58 ng/ml). Diurnal cortisol profile was regular in all patients. After Synacthen injection cortisol level rose in all groups to the similar values at 60 min. The same stimuli induced intensive <em>17OHP</em> secretion in group 1 (2.42+/-2.02 ng/ml) at 30 min statistically higher than in group 2 (1.46+/-0.95 ng/ml), (p=0.045). None had 21-hydroxysase defect. There were positive correlation between levels of <em>17OHP</em> and LH (r=0.38), <em>17OHP</em> and T (r=0.39), <em>17OHP</em> and LH/FSH (r=0.40) CONCLUSIONS: <em>17OHP</em> level in patients with PCOS is significantly higher then in other hirsute girls. High <em>17OHP</em> and normal cortisol level after Synacthen administration in PCOS girls point that activity of enzymes involved in <em>17OHP</em> production is augmented.

The effect of the antiestrogen tamoxifen (Tx) on the acute and chronic hCG administration was evaluated in patients with hypogonadotropic hypogonadism (HH) and in normal men. An hCG test (5000 IU hCG) was performed before, after two months of hCG administration (2000 IU hCG three times weekly) and after two months of hCG + Tx (2000 IU hCG three times weekly plus 20 mg/day of tamoxifen). Blood samples were obtained before and following 24 and 72 h of every test to determine T, E, <em>17OHP</em> and SHBG. T increased only in HH with both treatments (X +/- SEM: Basal: 97.9 +/- 19.7; hCG: 237.7 +/- 43.2; hCG +/- Tx: 204.7 +/- 10.7 ng/100 ml). <em>17OHP</em> rose with hCG alone, but not with hCG + Tx in both groups. E, SHBG and <em>17OHP</em>/T ratio did not change after treatments. hCG tests: E increased 24 h following hCG administration in every test. The ratio <em>17OHP</em>/T rose at 24 h in the first and second test but in the third test it did not change. These results support the role of E in the acute hCG-induced Leydig cell desensitization. However, the association of Tx does not improve T serum levels, suggesting that E might not be the unique factor involved in the mechanisms for testicular desensitization.

In Australia, all newborns born in New South Wales (NSW) and the Australia Capital Territory (ACT) have been offered screening for rare congenital conditions through the NSW Newborn Screening Programme since 1964. Following the development of the Australian Newborn Bloodspot Screening National Policy Framework, screening for congenital adrenal hyperplasia (CAH) was included in May 2018. As part of the assessment for addition of CAH, the national working group recommended a two-tier screening protocol determining 17α-hydroxyprogesterone (<em>17OHP</em>) concentration by immunoassay followed by steroid profile. A total of 202,960 newborns were screened from the 1 May 2018 to the 30 April 2020. A threshold level of <em>17OHP</em> from first tier immunoassay over 22 nmol/L and/or top 2% of the daily assay was further tested using liquid chromatography tandem mass spectrometry (LC-MS/MS) steroid profiling for <em>17OHP</em> (MS<em>17OHP</em>), androstenedione (A4) and cortisol. Samples with a ratio of (MS<em>17OHP</em> + A4)/cortisol > 2 and MS<em>17OHP</em> > 200 nmol/L were considered as presumptive positive. These newborns were referred for clinical review with a request for diagnostic testing and a confirmatory repeat dried blood spot (DBS). There were 10 newborns diagnosed with CAH, (9 newborns with salt wasting CAH). So far, no known false negatives have been notified, and the protocol has a sensitivity of 100%, specificity of 99.9% and a positive predictive value of 71.4%. All confirmed cases commenced treatment by day 11, with none reported as having an adrenal crisis by the start of treatment.

Keywords: 17-α hydroxyprogesterone; congenital adrenal hyperplasia; immunoassay; liquid chromatography tandem mass spectrometry; newborn screening; screening pathway.