BACKGROUND

The classical interpretation of the feedback regulation of the male hypothalamo-pituitary-gonadal axis predicts that a partial inhibition of testosterone (T) synthesis will result in a compensatory rise in LH secretion. The question arises as to whether such a compensation is complete or that decreased T synthesis may result in a lower plasma T concentration.

OBJECTIVE

To investigate whether a moderate inhibition of T synthesis capacity will be fully compensated by increased LH secretion. DESIGN, SUBJECTS AND INTERVENTIONS: In nine young healthy men, we partially inhibited T synthesis capacity using ketoconazole (KTZ) 100 mg four times daily. On day -6 (1 week prior to KTZ intake), days 1 and 8 of KTZ administration blood was drawn [07:00 h (t(1)), 10:00 h (t(2)), 13:00 h (t(3))] for evaluation of T, LH, oestradiol (E2), 17-OH-progesterone (<em>17OHP</em>), progesterone (PR) and sex hormone binding globulin (SHBG). On day 8, 5000 IU of hCG were administered to evaluate the maximal T secretion under KTZ.

RESULTS

Administration of KTZ resulted in an acute, moderate but significant decrease of plasma T concentration. On day 8, plasma LH, <em>17OHP</em> and PR were elevated relative to day -6 and day 1, but mean T was still lower compared to day -6. Mean E2 and SHBG were only slightly affected by KTZ. After stimulation by hCG, plasma T was restored to its baseline level.

CONCLUSIONS

These results argue against the assumption that a moderate decline in T synthesis capacity will be compensated completely by increased LH secretion.

OBJECTIVE

Liquid-chromatography tandem mass-spectrometry (LC-MS/MS) was developed in parallel to Immunoassays (IAs) and today is proposed as the "gold standard" for steroid assays. Leydig cells of men with Klinefelter syndrome (KS) are able to respond to human chorionic gonadotropin (hCG) stimulation, even if testosterone (T) production was impaired. The aim was to evaluate how results obtained by IAs and LC-MS/MS can differently impact on the outcome of a clinical research on gonadal steroidogenesis after hCG stimulation.

METHODS

A longitudinal, prospective, case-control clinical trial. (clinicaltrial.gov NCT02788136) was carried out, enrolling KS men and healthy age-matched controls, stimulated by hCG administration. Serum steroids were evaluated at baseline and for 5 days after intramuscular injection of 5000 IU hCG using both IAs and LC-MS/MS.

RESULTS

13 KS patients (36 ± 9 years) not receiving T replacement therapy and 14 controls (32 ± 8 years) were enrolled. T, progesterone, cortisol, 17-hydroxy-progesterone (<em>17OHP</em>) and androstenedione, were significantly higher using IAs than LC-MS/MS. IAs and LC-MS/MS showed direct correlation for all five steroids, although the constant overestimation detected by IAs. Either methodology found the same <em>17OHP</em> and T increasing profile after hCG stimulation, with equal areas under the curves (AUCs).

CONCLUSIONS

Although a linearity between IA and LC-MS/MS is demonstrated, LC-MS/MS is more sensitive and accurate, whereas IA shows a constant overestimation of sex steroid levels. This result suggests the need of reference intervals built on the specific assay. This fundamental difference between these two methodologies opens a deep reconsideration of what is needed to improve the accuracy of steroid hormone assays.

We studied the kinetics of testicular response to human chorionic gonadotropin (hCG) in oligoasthenospermic and asthenospermic patients (OAZ-AZ). The responses of testosterone (T), androstenedione (A), 17 OH-progesterone (<em>17OHP</em>), and estradiol (E2) were evaluated in 60 OAZ-AZ patients and compared to those of 10 normal men. The responses of T, A, and <em>17OHP</em> to hCG in the control group displayed a biphasic pattern with an initial peak at 4 hours and a second peak after 24 hours. The E2 response showed a single peak between 24 and 48 hours after hCG administration. OAZ-AZ patients had two types of T responses: group 1 (n = 40) had no first peak and group 2 (n = 20) had a normal response pattern. The response of A was similar to that of T, and the E2 response was normal in both groups. There were three types of <em>17OHP</em> responses in group 1 (low, high, or normal); however, the <em>17OHP</em> response was normal in group 2. Treatment of group 1 with aromatase inhibitors (aminoglutethimide or testolactone) induced an improvement of the acute T response only in patients with high or normal <em>17OHP</em> response to hCG, whereas no effects were observed in patients with low <em>17OHP</em> response. In group 2, the aromatase inhibitors induced no changes in the T response. These results demonstrate that in some OAZ-AZ patients (group 1, blunted T response) testicular hormone production is altered. They also suggest the presence of two enzyme blocks: one at the 17,20 desmolase level, mediated by E2, and another at early biosynthetic steps, not mediated by E2.

Samples of cord serum from 29 healthy neonates were analysed for digoxin-like immunoreactive substance (DLIS), cortisol, 17 beta-oestradiol, progesterone, dehydroepiandrosterone-sulphate (DHEAS), 17 alpha-hydroxyprogesterone (<em>17OHP</em>), androstenedione, oestriol and ouabain-like activity (OLA; by inhibition of Na+, K+ATPase activity). The mean serum concentration of DLIS was 0.91 (SD = 0.19) nmol/L and the mean OLA was 26.1 (SD = 11.5) nmol/L. There was no correlation between DLIS and OLA. DLIS correlated significantly with oestriol (r = 0.521), progesterone (r = 0.534) and <em>17OHP</em> (r = 0.43). Stepwise multiple regression analysis showed that 17 beta-oestradiol, progesterone and androstenedione contributed to DLIS and the intercept was 0.64 (SD = 0.127). The concentrations of steroids (17 beta-oestradiol, progesterone, androstenedione) required to displace digoxin by 50% in the digoxin immunoassay and inhibit Na+,K+ATPase in the OLA assay were 10(3)-10(4)-fold higher than those found in cord serum. We conclude that the contribution of these steroids to DLIS is small and that DLIS and OLA measure different compounds.

OBJECTIVE

To investigate the role of adjuvant 17-α-hydroxy-progesterone caproate (<em>17OHP</em>-C) in reducing the risk of preterm delivery <34 weeks and adverse perinatal outcomes in women with ≥3 second trimester pregnancy losses attributed to cervical insufficiency undergoing prophylactic cerclage.

METHODS

Retrospective cohort study of women with prophylactic cerclage placed between 2006 and 2014 divided into a cohort of (i) those receiving adjuvant <em>17OHP</em>-C (n=43), and (ii) controls with cerclage alone (n=59).

RESULTS

Demographic characteristics were comparable in both groups. There was no significant difference in gestational age at delivery between the cerclage-<em>17OHP</em>-C group (33.4±5.6 weeks) and the cerclage-alone group (34.4±4.6 weeks); P=0.33. We noted a non-significant increase for deliveries <34 weeks in the cerclage-<em>17OHP</em>-C group (44.2%) compared to controls (28.8%) which remained non-significant after adjusting for confounders; P=0.46. There was no statistically significant difference in the rate of delivery <37, 32, 28 and 24 weeks. Adverse neonatal outcomes were comparable in both groups (cerclage-<em>17OHP</em>-C 48.8% vs. cerclage-alone 39%); P=0.43.

CONCLUSIONS

Intramuscular <em>17OHP</em>-C in combination with prophylactic cerclage in women with cervical insufficiency and ≥3 second trimester pregnancy losses had no synergistic effect in reducing the rate of recurrent preterm birth or improving perinatal outcomes.

An indirect enzyme immunoassay for the measurement of total 17alpha-hydroxyprogesterone (<em>17OHP</em>) in serum using monoclonal antibodies generated in our laboratory was developed. Here, (a) instead of extraction with solvents, serum was heated to free protein-bound <em>17OHP</em> and assay was performed at pH 9.6, (b) to ensure uniform assay conditions for both standards and samples, buffer for standards contained charcoal-stripped pre-heated pooled cord serum. Assays were done in 96-well EIA microplates pre-coated with 17alpha-hydroxyprogesterone-3-(o-carboxymethyl)oxime: bovine serum albumin. Secondary antibody was horseradish peroxidase-linked sheep anti-mouse IgG polyclonal antibody. The method was accurate and suitable for screening for congenital adrenal hyperplasia.

OBJECTIVE

To assess serum 17-alpha-hydroxyprogesterone (<em>17OHP</em>) and corticosterone concentrations in dogs with nonadrenal neoplasia and dogs being screened for hyperadrenocorticism.

METHODS

Prospective study.

METHODS

16 clinically normal dogs, 35 dogs with nonadrenal neoplasia, and 127 dogs with suspected hyperadrenocorticism.

METHODS

ACTH stimulation tests were performed in all dogs. Baseline serum cortisol and corticosterone concentrations were measured in the healthy dogs; baseline serum cortisol concentration and ACTH-stimulated cortisol, corticosterone, and <em>17OHP</em> concentrations were measured in all dogs. Endogenous plasma ACTH concentration was also measured before administration of ACTH in dogs with neoplasia.

RESULTS

In 35 dogs with neoplasia, 31.4% had high serum <em>17OHP</em> concentration and 22.9% had high serum corticosterone concentration. Of the 127 dogs with suspected hyperadrenocorticism, 59 (46.5%) had high ACTH-stimulated cortisol concentrations; of those, 42 of 59 (71.2%) and 32 of 53 (60.4%) had high serum <em>17OHP</em> and corticosterone concentrations, respectively. Of dogs with serum cortisol concentration within reference range after ACTH administration, 9 of 68 (13.2%) and 7 of 67 (10.4%) had high serum <em>17OHP</em> and corticosterone concentrations, respectively. In the dogs with neoplasia and dogs suspected of having hyperadrenocorticism, post-ACTH serum hormone concentrations were significantly correlated.

CONCLUSIONS

Serum concentrations of <em>17OHP</em> or corticosterone after administration of ACTH may be high in dogs with nonadrenal neoplasia and no evidence of hyperadrenocorticism. Changes in serum <em>17OHP</em> or corticosterone concentrations after administration of ACTH are proportionate with changes in cortisol concentration.

Unexplained hyperandrogenic oligoanovulation is a main feature of polycystic ovary syndrome (PCOS). P450c17 phosphorylation selectively increases 17,20-lyase activity and androgen biosynthesis but minimally affects 17α-hydroxylase. Studies have recently identified mitogen-activated protein kinase 14 (MAPK14, p38α) as the kinase responsible for enhancing 17,20-lyase activity through P450c17 phosphorylation. We investigated whether oxidant-induced oxidative stress increases 17,20-lyase activity through oxidant-sensitive p38α signaling pathways. NCI-H295R adrenal cells were treated with three oxidants, palmitate, H₂O₂ and 4-hydroxy-2-nonenal (HNE), to simulate the excessive oxidative stress of PCOS. Oxidant exposure significantly induced dehydroepiandrosterone production and increased p38α phosphorylation and activation, but the effect on 17α-hydroxyprogesterone production was far less clear. None of the treatments altered the expression of P450c17 or its necessary factors POR and b5. LC-MS/MS revealed increased DHEA production in NCI-H295R cells. Both p38α inhibition and siRNA-mediated silencing attenuated H₂O₂- or 0.45-0.75 mM PA-mediated augmentation of DHEA production with relatively stable <em>17OHP</em> levels, indicating that activated p38α mediates oxidative stress-induced 17,20-lyase activation and androgen synthesis stimulation, which may underlie hyperandrogenism in PCOS.

In a chronic inflammatory disease, such as rheumatoid arthritis (RA), the hypothalamic-pituitary-adrenal axis is altered in three ways: (1) the inflammation-related spontaneous and stimulated secretion of cortisol is inadequate; (2) the inflammation-related secretion of adrenocorticotropic hormone (ACTH) is low; and (3) the levels of adrenal androgens decrease. In patients with RA, long-term therapy with anti-TNF therapy sensitizes the pituitary gland and improves adrenal androgen secretion. We have recently found that the mean serum levels of ACTH, cortisol, 17-hydroxyprogesterone (<em>17OHP</em>), and androstenedione (ASD) in 11 prednisolone-naïve patients with psoriatic arthritis did not markedly change during 12 weeks of etanercept treatment, nor did the serum cortisol/ACTH ratio. However, the greater increase in serum cortisol in comparison with serum <em>17OHP</em> or ASD was related to clinical improvement, which indicates that the improvement was more related to the higher cortisol levels.

21-Hydroxylase deficiency (21OHD) is the commonest form of congenital adrenal hyperplasia, while 11betaOHD represents 5% of cases. Although both result from mutations in distinct genes, cases of 'apparent' combined 21OHD and 11betaOHD (AC21,11OHD) have been occasionally reported. A 6 year-old girl, born with ambiguous genitalia and salt-loss, had serum elevations (ng/dl) of androstenedione (>1,000), 17-hydroxyprogesterone (<em>17OHP</em>; 38,483), 21-deoxycortisol (21DF; 23,338), and 11-deoxycortisol (S; 4,928), suggesting AC21,11OHD. CYP21A and CYP11B1 genotyping identified mutations only in the former. On follow-up, serum S became normal but <em>17OHP</em> and 21DF were still elevated. ACTH stimulation disclosed elevated levels of <em>17OHP</em> and 21DF, but unresponsive S and undetectable deoxycorticosterone. The hormonal pattern initially suggested AC21,11OHD, but subsequent normalization of S showed transient 11-hydroxylase inhibition. This may have occurred by enzyme or co-enzyme immaturity or functional discrepancy, but also by selective inhibition of 11betaOH by excess intra-adrenal concentration of androgens, acting as pseudo-substrates for this enzyme.

<AbstractText>Chronic central serous chorioretinopathy (cCSC) is characterized by fluid accumulation between photoreceptors and the retinal pigment epithelium of which the cause is unknown. Associations with steroid use, stress, pregnancy, and the male sex suggest a role for the steroid hormone system in the disease. Here, we performed a comprehensive analysis of the steroid hormone system in active cCSC.</AbstractText><AbstractText>Serum hormone levels of 17 steroid hormones were measured in 46 male Caucasian patients with active cCSC and 46 male Caucasian age-matched controls using the AbsoluteIDQ stero17 kit.</AbstractText><AbstractText>Elevated levels of androsterone, estrone, etiocholanolone, and androstenedione were observed in cCSC patients compared with controls. Median hormone levels in cCSC patients versus controls, respectively, were as follows: androsterone, 0.84 ng/mL (interquartile range [IQR] = 0.61-1.06) versus 0.69 ng/mL (IQR = 0.48-0.96, P = 0.031); estrone, 0.12 ng/mL (IQR = 0.10-0.15) versus 0.10 ng/mL (IQR = 0.08-0.11; P = 0.0048); etiocholanolone, 0.19 ng/mL (IQR = 0.15-0.29) versus 0.13 ng/mL (IQR = 0.099-0.20, P = 0.0061). Mean levels of androstenedione were 3.10 ng/ml (SD = 1.03) versus 2.55 ng/mL (SD = 0.95), in cCSC patients versus controls, respectively. Additionally, Spearman's correlations between aldosterone and 11-deoxycortisol, androsterone, DHEA, DHEAS, and E1 differed between cCSC patients and controls, as well as between andosterone and E1, and between DHT and <em>17OHP</em>.</AbstractText><AbstractText>We present a comprehensive overview of the status of the steroid hormone system in active cCSC. Levels of four hormones were elevated in cCSC patients compared with controls, and the relationships between steroid hormones was altered, indicating that the balance in the steroid hormone system is altered in cCSC patients.</AbstractText>

BACKGROUND

Polycystic ovary syndrome (PCOS) and congenital adrenal hyperplasia (COH) are heterogeneous disorders, in which excess of androgens may be caused by improper function of ovaries and/or adrenals. In many cases an overlap between ovarian and adrenal type of functional hyperandrogenism has been observed. The relationship between adrenal and ovarian metabolism in hyperandrogenic women is not totally known and etiologic diagnosis of female hyperandrogenism is often difficult. The aim of the present study was to evaluate the usefulness of combined Dexamethasone-Triptoreline testing in distinguishing ovarian and adrenal type of functional hyperandrogenism, and checking if the test could be shortened in order to economise it.

METHODS

We have examined 57 women with androgen excess divided into two groups: ovarian (n = 42) and adrenal (n = 15) and 20 women with idiopathic hirsutism. There was also one patient suffering from Morris syndrome taken under examination just for curiosity. The blood for hormonal assay was taken in baseline conditions at 8.00 a.m. for LH, FSH, PRL, cortisol, T, DHEAS, <em>17OHP</em>, E2. Dx was given for 4 days 0.5 mg p.o. every 6 hours. 8 hours after the last Dx administration, the blood was taken for <em>17OHP</em> and T. Immediately after that Triptorelin 100 mg was given s.c. Then the blood was collected every 4 hours during 24 hours for <em>17OHP</em> estimation.

RESULTS

Decrease in T levels (from 1.65 +/- 0.52 to 0.73 +/- 0.25 ng/ml) after Dexamethasone administration was observed in adrenal group, which indicates adrenal glands as a source of excessive androgen production. No significant differences were seen in ovarian group. But in women from ovarian group supranormal <em>17OHP</em> response after Triptoreline administration was seen: (ng/ml): at 8.00 am-0.68 +/- 0.44, 12.00--1.21 +/- 0.7*, 16.00--1.71 +/- 1.19*, 20.00--2.39 +/- 1.81*, 24.00--3.41 +/- 2.64*, 4.00--3.91 +/- 2.82*, 8.00--6.06 +/- 2.43* (*p < 0.01, **p < 0.001). Such a response is typical for women with well defined PCOS and other forms of functional ovarian hyperandrogenism and indicates ovary as a source of androgens. Significant differences were also noticed in idiopathic group: 8.00--0.31 +/- 0.09, 12.00--0.38 +/- 0.17, 16.00--1.41 +/- 0.62*, 20.00--1.52 +/- 0.97*, 24.00--1.89 +/- 0.83*, 4.00--2.17 +/- 0.83*, 8.00--1.83 +/- 0.71** (*p < 0.01). <em>17OHP</em> levels did not change significantly during the whole test in adrenal group: 8.00--1.83 +/- 1.24, 12.00--1.91 +/- 1.37, 16.00--1.95 +/- 0.86, 20.00--2.19 +/- 0.93, 24.00--2.63 +/- 1.58, 4.00--2.56 +/- 1.78, 8.00--237 +/- 0.94. But patients from this group had exaggerated <em>17OHP</em> response to ACTH (from 4.32 +/- 1.31 to 15.34 +/- 4.1 ng/ml). In patient suffering from Morris syndrome, after Triptoreline, serum <em>17OHP</em> levels reminded on the same level as they were before drug administration.

CONCLUSIONS

Combined Dx-Triptorelin test can be very useful to distinguish ovarian and adrenal type of functional hyperandrogenism. The number of times of blood collection for <em>17OHP</em> can be reduced to 4 times a day (during 24 hours): at 8.00, 20.00, 24.00, 8.00.

Objective: To evaluate the diagnostic value of medium dose dexamethasone androgen suppression tests (DAST) in female hyperandrogenism. Methods: DAST results were retrospectively analyzed in 85 cases of women with hyperandrogenism including 55 cases of congenital adrenal hyperplasia (CAH), 10 cases of testosterone-producing tumors and 20 cases of polycystic ovary syndrome (PCOS) between January 1984 and December 2017 in Peking Union Medical College Hospital. The suppression rate of testosterone and 17 hydroxyprogesterone (<em>17OHP</em>) were evaluated. The cut-off point of suppression rates were calculated by receiver operating characteristic (ROC) curve in the differential diagnosis of CAH and non-CAH causes. Results: The 1-day medium dose DAST was performed simultaneously in 41 cases of CAH patients and the 5-days medium dose DAST was performed simultaneously in 19 cases of CAH patients. The results indicated that the average suppression rate of testosterone were 77.9% and 91.3% (P<0.001) and the average suppression rate of <em>17OHP</em> was 95.2% and 97.0%, respectively (P=0.220). In patients (41 cases of CAH, 10 cases of testosterone producing tumor and 20 cases of PCOS) with 1-day DAST, the optimal testosterone suppression rate was 61.2% (the sensitivity and specificity was 87.8% and 96.7%, respectively) and the optimal <em>17OHP</em> suppression rate was 87.1% (the sensitivity and specificity was 95.1% and 93.3%, respectively) in the identification of CAH and non-CAH cases. There is no clinical significance between the testosterone and <em>17OHP</em> suppression rate in the differential diagnosis of CAH and non-CAH cases. Conclusions: There was no difference in the suppression rate of <em>17OHP</em> between the 1-day and 5-days DAST in CAH cases. The sensitivity of suppression rate of <em>17OHP</em> is equal in the differential diagnosis of hyperandrogenism. One-day approach DAST could be used as functional test for the diagnosis of the etilology of hyperandrogenism (CAH or non-CAH).

OBJECTIVE

To investigate the pituitary and adrenal hormone response after an intravenous epinephrine challenge in patients with rheumatoid arthritis (RA) and controls.

METHODS

Fifteen untreated female patients with RA (age 51.5 +/- 3.2 yrs) and 7 healthy female controls (48.0 +/- 4.3 yrs) were infused with epinephrine (0.05 microg/kg/min) for about 20 min. Plasma levels of adrenocorticotropic hormone (ACTH), and serum levels of cortisol, 17-hydroxyprogesterone (<em>17OHP</em>), and dehydroepiandrosterone sulfate (DHEAS) were analyzed at baseline and shortly after cessation of epinephrine infusion (20 min).

RESULTS

At baseline and after epinephrine infusion, serum levels of cortisol (p = 0.045) and <em>17OHP</em> (p = 0.021) were higher in controls compared to patients with RA. In contrast, at baseline and after epinephrine infusion, plasma levels of ACTH and serum levels of DHEAS were similar in controls and patients. After epinephrine infusion, only the patients with RA had a significant decrease of serum cortisol (p = 0.026) and serum <em>17OHP</em> (p = 0.026). Plasma levels of ACTH (p = 0.073) and serum levels of DHEAS (p = 0.055) tended to decrease.

CONCLUSIONS

Serum cortisol and <em>17OHP</em> (cortisol precursor) were lower in patients with RA compared to controls despite similar ACTH levels. Simulation of an adrenomedullary stress response by epinephrine infusion decreased serum cortisol and <em>17OHP</em> in patients but not in controls. Such a response may play an unfavorable role during a typical stress reaction in patients with RA that may lead to a more proinflammatory situation.

To learn how progesterone (P) inhibits follicle growth during the luteal phase, we determined whether P will inhibit follicle growth when follicle-stimulating hormone (FSH) is secreted in large amounts, namely, after luteectomy (CLX) in monkeys with only one ovary. Second, a functional role for 17 alpha-hydroxyprogesterone (<em>17OHP</em>) was examined as a common mediator of the inhibition of folliculogenesis by the dominant follicle and corpus luteum. To accomplish the first goal, nine chronically hemiovarectomized monkeys were lutectomized chronically hemiovariectomized monkeys were luteectomized at midluteal phase. In five monkeys that received no steroid, the next preovulatory luteinizing hormone (LH) surge occurred 14.0 +/- 0.8 days (mean +/- SE) after CLX. In contrast, the next LH surge was delayed in four monkeys implanted for 10 days with Silastic capsules containing P and occurred 25.0 +/- 2.7 days after CLX, i.e., 14.8 +/- 2.7 days after the capsule removal. In both groups, FSH levels increased markedly after CLX to a comparable degree and duration; yet, only a single follicle ovulated in each monkey. To examine a potential inhibitory role for <em>17OHP</em>, monkeys with two ovaries were luteectomized and received 1) no steroid, 2) <em>17OHP</em> via Silastic capsules, or 3) P for 10 days after CLX. Progesterone replacement after CLX appeared to maintain <em>17OHP</em> levels, which showed a transient decrease after CLX alone. As above, P delayed the next LH surge (25.4 +/- 1.3 vs. 15.0 +/- 0.6 days) despite comparable increases in serum FSH after CLX alone. Replacement at two levels of <em>17OHP</em> did not delay the onset of menses (2-3 days post-CLX) or significantly delay the next LH surge 18.3 +/!- 1.9 or 20.8 +/- 3.4 vs. 15.0 +/- 0.6 days (P greater than 0.2) in monkeys CLX only. Whatever may be the mode of action of P, it appears that it is not mediated by peripheral conversion to <em>17OHP</em>. These findings demonstrate that P at luteal phase levels can inhibit follicle growth culminating in ovulation even in the face of sustained, elevated levels of endogenous FSH. Because single ovulations occurred despite unambiguous and prolonged increments in serum FSH after CLX, the precise regulation of the ovulatory quota in this primate appears to be accomplished by means other than FSH alone.

The most significant action of progesterone appears to be on the cervix and in prevention rather than on treatment of preterm delivery. In women with singleton gestations, no prior PTB, and CL <20 mm at <24 weeks, vaginal progesterone, either 90 mg gel or 200 mg suppository, is associated with reduction of both preterm birth (PTB) and perinatal morbidity/mortality. Cerclage is as effective as vaginal progesterone in women with CL <25 mm. Treatment of women with previous PTB with <em>17OHP</em>-C from 16 to 20 weeks' gestation until 36 weeks could reduce significantly both the risk of delivery at <37, <35 and <32 weeks' gestation, as well as the rates of NEC, the need for supplemental oxygen and IVH. In women successfully treated with tocolytics progesterone combined with corticosteroid therapy lengthens pregnancy, reduces occurrence of respiratory distress syndrome and low birth weight. However, there is currently insufficient evidence on the role of progesterone after arrested preterm labor. It is reasonable to support an approach with CL screening of women with prior PTB starting at 16 to 19 weeks and administration of progesterone to women with a short cervix. Cerclage may be offered to those with a CL<25 mm. A combination of traditional tocolytics, corticosteroids and progesterone might be beneficial.

The aim of the present study was to evaluate and compare the response of 17 OHP to ACTH stimulation in patients with various types of adrenal incidentalomas and to examine the occurence of germline CYP21 mutation in these patients.

METHODS

40 patients (27 females, 13 males) with unilateral and bilateral masses were screened for fi ve most common mutations of the CYP21 in peripheral blood DNA samples. A hormonal evaluation, i.e. baseline plasma values of <em>17OHP</em>, DHEAS as well as plasma <em>17OHP</em> and DHEA after ACTH stimulation, was performed in all patients. 21 of them had unilateral adrenal adenoma, 13 patients had adrenal hyperplasia (six of them unilateral) and 6 patients had CT characteristics of other tumors (myelolipomas, cysts, adrenocortical carcinoma).

RESULTS

There were no significant differences in plasma <em>17OHP</em>, DHEAS and plasma cortisol between all three groups. Stimulated plasma values of DHEA and <em>17OHP</em> after ACTH administration were significantly higher in patients with adenomas (p < 0.05 and p < 0.01) and with hyperplasia (p < 0.05 and p < 0.05) compared with those with other tumors. An exaggerated response of 17 OHP was found in 5 (12 % ) patients. However, mutation screening in peripheral blood samples revealed no CYP21 mutation in all examined groups.

CONCLUSIONS

Although 12 % of patients with adrenal incidentalomas had an exaggerated response of 17 OHP after ACTH administration indicating a possible 21-hydroxylase deficiency, these findings are not associated with CYP21 mutation estimated in peripheral blood samples. There was found no germline CYP21 mutation in all patients with various adrenal incidentalomas.

Serum sulphates of 5-androstene-3 beta,17 beta-diol (5-ADIOL-S), 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-DIOL-S) and dehydroepiandrosterone (DHEA-S), unconjugated androstene-dione (AD) and testosterone (T), sex hormone binding globulin (SHBG), free androgen index (FAI), 17 alpha-hydroxyprogesterone (<em>17OHP</em>), luteinising hormone (LH) and follicle stimulating hormone (FSH) were measured by specific radioimmunoassay in 28 hirsute women with polycystic ovarian disease (PCO) and in normal women (n = 73). Mean levels of steroids measured were significantly elevated, and SHBG significantly depressed, in the women with PCO with values (mean +/- SE) for 5-ADIOL-S (516 +/- 51 vs 267 +/- 10 nmol/l), 3 alpha-DIOL-S (130 +/- 9 vs 52 +/- 2 nmol/l), DHEA-S (7.3 +/- 0.5 vs 4.4 +/- 0.2 mumol/l), AD (11.3 +/- 1.1 vs 3.4 +/- 0.2 nmol/l), T (3.3 +/- 0.2 vs 1.5 +/- 0.1 nmol/l) and <em>17OHP</em> (5.1 +/- 0.8 vs 2.8 +/- 0.2 nmol/l). SHBG levels were 31 +/- 2.9 vs 65 +/- 2.5 nmol/l, and the free androgen index [100 x T (nmol/l) divided by (SHBG nmol/l)] was 12.5 +/- 1.4 vs 2.4 +/- 0.1. The mean LH to FSH ratio was also elevated at 2.8 +/- 0.3. These studies suggest that the measurement of 5-ADIOL-S and DHEA-S may indicate adrenal gland involvement in PCO while 3 alpha-DIOL-S appears to be a reflection of peripheral androgen metabolism. A comprehensive biochemical profile of PCO should thus include the analysis of these sulphoconjugates as well as unconjugated steroids.

Nonflagellated germ cells were isolated from rainbow trout testis to determine their ability to synthesize 17,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta OHP), a progestin involved in the control of the release of sperm. Germ cells were obtained by enzymatic dissociation (collagenase; 3 mg.ml-1, 4.5 h, 12 degrees C) from testes that were immature and at the beginning of spermatogenesis. Somatic cells were eliminated by adhesion to the culture plates. Dose-related amounts of 17,20 beta OHP were measured by RIA in culture media of germ cells incubated with increasing dosages of 17-hydroxyprogesterone (<em>17OHP</em>; 0.05-10 micrograms.ml-1) for 20 h at 12 degrees C. Furthermore, 3H-17,20 beta OHP was identified by chromatography and co-crystallization with a reference in incubating cells provided by 3H-<em>17OHP</em> (2.5 and 4 h, 12 degrees C). Other metabolites were detected but not identified. 11-Ketotestosterone (11KT) was either nondetectable by RIA in control cultures or, when detected, was found at very low levels. In no case was 11KT stimulated by addition of <em>17OHP</em> or gonadotropin II (GtH II; 400 ng.ml-1); this indicated the absence of contamination by Leydig cells. Thus, to our knowledge, this report demonstrates steroidogenic activities in nonflagellated germ cells of fish testis for the first time. 20 beta-Hydroxysteroid dehydrogenase (20 beta HSD) activity was identified, showing that germ cells are able to synthesize 17,20 beta OHP at an early stage in rainbow trout testis.

3β-hydroxysteroid dehydrogenase II (3β-HSD) deficiency represents a rare CAH variant. Newborns affected with its classic form have salt wasting in early infancy and genital ambiguity in both sexes. High levels of 17-hydroxypregnenolone (Δ5<em>17OHP</em>) are characteristic, but extra-adrenal conversion to 17-hydroxyprogesterone (<em>17OHP</em>) may lead to positive results on newborn screening tests. Filter paper <em>17OHP</em> on newborn screening test was performed by immunofluorometric assay, and serum determinations of <em>17OHP</em> and Δ5<em>17OHP</em>, by radioimmunoassay. A 46,XY infant with genital ambiguity and adrenal crisis at three months of age presented a positive result on newborn screening for CAH. Serum determinations of <em>17OHP</em> and Δ5<em>17OHP</em> were elevated, and a high Δ5<em>17OHP</em>/cortisol relation was compatible with the diagnosis of 3β-HSD deficiency. Molecular analysis of the HSD3B2 gene from the affected case revealed the presence of the homozygous p.P222Q mutation, whereas his parents were heterozygous for it. We present the first report of 3β-HSD type II deficiency genotype-proven detected at the Newborn Screening Program in Brazil. The case described herein corroborates the strong genotype-phenotype correlation associated with the HSD3B2 p.P222Q mutation, which leads to a classic salt-wasting 3β-HSD deficiency. Further evaluation of <em>17OHP</em> assays used in newborn screening tests would aid in determining their reproducibility, as well as the potential significance of moderately elevated <em>17OHP</em> levels as an early indicator to the diagnosis of other forms of classic CAH, beyond 21-hydroxylase deficiency.

Critical issue regarding to variation of findings based on different phenotypes led investigators to define whether they are distinct features or overlapping ones. Therefore, we aimed to investigate the association between diverse phenotypes of PCOS (Poly Cystic Ovary Syndrome) with clinical manifestations, anthropometric indices, and metabolic characteristics. This was a descriptive cross-sectional study conducted in 15-39 years old women with PCOS referred to infertility clinics in the north part of Iran, Rasht during 2010-2011. Data were gathered through an interview by a form consisted of demographic characteristics, laboratory findings, ovarian volume and anthropometric indices. A total of 214 patients consisted of 161 PCOS (cases) and 53 normal women (controls) participated in this study. The most prevalent phenotype in PCOS population was IM/PCO/HA (54%), followed by IM/HA (28%) and IM/PCO (13%). PCO/HA was present only in 6 PCOS patients (5%). PCOS patients were significantly younger than controls (P=0.07). Results showed that increased ovarian volume were higher in PCOS group in comparison with controls and IM/PCO/HA, and IM/PCO had respectively the largest ovarian volumes. Also, a significant relation was observed based on Cholesterol, <em>17OHP</em>, LH, TG, 2hpp, and LH/FSH between patients with PCOS and control groups. There were significant differences in demographic, anthropometric, hormonal and ultrasound findings between PCOS and controls. Therefore, it seems that classification of the characteristics of each phenotype could offer an appropriate guide for screening risks of PCOS and may facilitate performing most favorable treatment for these complications.

The electrophoretic profiles of the proteins in the serum and cerebrospinal fluid (CSF) of the rhesus monkey were studied by polyacrylamide gel electrophoresis. The association of tritium labelled estradiol (3H-E), progesterone (3H-P) and 17 alpha-hydroxyprogesterone (3H-<em>17OHP</em>) with these proteins was studied under in vitro and in vivo conditions. The CSF was found to contain, besides albumin, at least 4 globulins with similar electrophoretic mobilities as those found in the serum. The electrophoretic mobilities of these globulins were Ralb 0.4, 0.6, 0.8 and 0.9. The electrophoretic profiles of the monkey serum and CSF proteins were found to be similar to those described for human serum and CSF. 3H-E and 3H-P were respectively associated with serum globulins of Ralb 0.4 and 0.8 both in the in vitro and in vivo studies. The respective Ralb of these 2 globulins were similar to the testosterone-estradiol binding globulin and cortisol binding globulin in human serum. 3H-<em>17OHP</em> was found to be associated with several globulins under both in vitro and in vivo conditions. In marked contrast to the in vitro studies, radioactive products were found to be associated with albumin in vivo. It is presumed that these products may be steroidal conjugates derived from the systemic metabolism of the administered steroids. No radioactivity was associated with the CSF protein after in vitro incubation with any of the three 3H-steroids. In the in vivo studies, however, radioactivity was associated with such of the CSF proteins that were similar to those found in the serum and with which the administered steroids were associated.

Pituitary hormones, adrenal and testicular steroids and steroid-binding proteins were analysed in 4 active ice hockey players and in 3 spectators, matched for physical fitness, during the night after a 26-hour cup tournament. The examined period included the last match of three matches. Unconjugated adrenal steroids (cortisol, 4-androstene-3,17-dione [A-4] and dehydroepiandrosterone [DHA]) and prolactin were increased in players immediately following the match, but were normalized within 3-4 h. After that similar nocturnal patterns were observed in players and spectators. The time-courses for DHA sulphate, albumin and sex hormone binding globulin (SHBG) were identical in players and spectators. Serum testosterone (T), 17 alpha-hydroxyprogesterone (<em>17OHP</em>) and LH levels had time-courses with a nadir between 20.00 and 24.00 h and maximum values in the morning; however, this pattern was less pronounced in players. The results illustrate the rapid normalization of adrenocortical activity after physical stress and further support the view that interference with pituitary LH secretion may be the main mechanism responsible for the observed changes in testicular steroids.

OBJECTIVE

To determine concentrations of 17alpha-hydroxyprogesterone (<em>17OHP</em>) in serum of healthy bitches during various stages of the reproductive cycle and in bitches with hyperadrenocorticism and to compare the dynamics of <em>17OHP</em> with those of progesterone.

METHODS

Prospective evaluation study.

METHODS

15 healthy sexually intact bitches and 28 spayed bitches with hyperadrenocorticism.

METHODS

11 healthy bitches were evaluated during estrus, nonpregnant diestrus, and anestrus (group 1); 4 other healthy bitches were evaluated during pregnancy and after ovariohysterectomy (group 2). Cycle stages were determined via physical examination, vaginal cytologic evaluation, and serum progesterone concentration. Bitches with hyperadrenocorticism were evaluated once at the time of diagnosis (group 3). Serum hormone concentrations were determined with immunoassays.

RESULTS

In group 1, the serum <em>17OHP</em> concentration was significantly higher in diestrus (median, 1.8 ng/mL) than in estrus (median, 1.1 ng/mL) and anestrus (median, 0.2 ng/mL) and higher in estrus than in anestrus. Changes in serum progesterone concentrations accounted for 22% (estrus) or 23% (diestrus) of the variation in serum <em>17OHP</em> concentrations. In group 2, <em>17OHP</em> and progesterone concentrations were significantly higher during pregnancy than after ovariohysterectomy. The serum <em>17OHP</em> concentration in group 3 was significantly lower (median, 0.2 ng/mL) than in group 1 in estrus and diestrus and in group 2 during pregnancy (median, 0.7 ng/mL) but was not different from <em>17OHP</em> concentrations in anestrus or after ovariohysterectomy (median, 0.2 ng/mL).

CONCLUSIONS

Serum <em>17OHP</em> concentrations in healthy bitches increased during estrus, diestrus, and pregnancy and at those times were higher than in spayed bitches with hyperadrenocorticism.