Dehydroepiandrostenedione Sulphate Research Articles

Androgens in women: Establishing reference intervals for dehydroepiandrostenedione sulphate and androstenedione on the Roche Cobas.

Immunoassays are the most common method in routine practice for measuring androgens in women. Study's aim was to establish new population specific indirect reference intervals (RI) for dehydroepiandrostenedione sulphate (DHEAS) and for new androstenedione test available on automated Roche Cobas electrochemiluminescent immunoassay method. From extracted laboratory records, testosterone, sex hormone binding globulin and follicle-stimulating hormone were used as reference tests to exclude possibly diseased women. After the data selection steps, the study included 3500 subjects for DHEAS and 520 for androstenedione aged 20-45 years. To evaluate the need for age partitioning, we calculated standard deviation ratio and bias ratio. For each hormone, 90% and 95% RIs were calculated with appropriate statistical method. Total age group (20-45 years) 95% RIs were: 2.77-11.50 µmol/L for DHEAS and 2.48-8.89 nmol/L for androstenedione. Age-stratified 95% RIs for DHEAS were: 3.65-12.76 µmol/L (20-25 years); 2.97-11.50 µmol/L (25-35 years) and 2.30-9.83 µmol/L (35-45 years). Age-stratified 95% RIs for androstenedione were: 3.02-9.43 nmol/L (20-30 years) and 2.23-7.75 nmol/L (30-45 years). New RIs for DHEAS were slightly wider for age group 20-25 and 35-45, while the differences in the age group 25-35 years were more pronounced. Androstenedione RI showed significantly higher concentrations than the manufacturer's. Age-related decrease of androgens should be considered when calculating RIs. We propose population specific, age-stratified RIs for DHEAS and androstenedione on electrochemiluminescent method, which should improve test interpretation in women of reproductive age.

Assessment of adrenal function in pediatric cancer survivors.

Oncological therapy can temporarily or permanently disrupt adrenal gland function. The aim of our study was to assess the function of adrenal glands in cancer survivors and to find the best diagnostic tools for it. Sixty patients aged 1.2-14.9 years (mean 8.3 ±3.5) with diagnosed malignancies and 45 healthy children as controls were recruited to the study. Patients were assessed 0-8 years (mean 2.4 ±2.0 years) after the oncological therapy. In all patients fasting blood samples were collected to measure: glucose, sodium, potassium, cortisol, aldosterone, plasma renin activity (PRA), dehydroepiandrostenedione-sulphate (DHEA-S), adrenocorticotropic hormone (ACTH) and antibodies against the adrenal cortex (AAA). Moreover, 24-hour urinary free cortisol (UFC) was assessed. Test with synthetic ACTH was carried out with 250 µg in neuroblastoma and nephroblastoma patients and with 1 µg in other oncological patients. The levels of morning cortisol and sodium were significantly lower and blood glucose were higher in cancer survivors than in controls (p = 0.006, p = 0.043, p = 0.008). Basal laboratory tests confirmed adrenal insufficiency (AI) in 1 patient with neuroblastoma. Low-dose ACTH revealed AI in 3 patients with acute lymphoblastic leukemia. In the study group, UFC correlated with evening and midnight cortisol (p = 0.001, p = 0.006). In the control group UFC correlated with DHEA-S (r = 0.623, p = 0.0001). None of assessed parameters correlated with the time since the completion of oncological therapy. The study confirmed possibility of developing asymptomatic AI in cancer survivors even several years after therapy. Instead of morning cortisol, classical diagnostic low-dose ACTH test seems to be an optimal tool for adrenal function's assessment.

Effect of Metformin on Anthropometric Measurements and Hormonal and Biochemical Profile in Patients with Prediabetes.

Objectives Metformin is the most widely preferred first-line oral antidiabetic agent that results in clear benefits in blood sugar regulation and diabetes-related complications. This study is aimed at assessing the effect of metformin on anthropometric, hormonal, and biochemical parameters in patients with prediabetes or insulin resistance. Methods A prepoststudy was conducted among 52 patients with prediabetes or insulin resistance who met the inclusion criteria. Weight, body mass index (BMI), and waist circumference were measured before and 12 months after metformin treatment. Serum concentrations of sex steroids, gonadotropins, and lipids were also assessed. Homeostasis model assessment (HOMA) index and quantitative sensitivity check (QUICKI) index scores were calculated before metformin treatment and after 12 months of use. Results After 12 months of metformin treatment, female patients had significant reduction in weight, BMI, and waist circumference after adjusting for age. Metformin use for 12 months resulted in significant reduction in mean fasting blood glucose and HbA1c in females only. Total cholesterol decreased significantly among men only and serum HDL-C showed a significant rise among females only. Serum LDL-C and triglycerides did not change significantly in females and males. Our study did now significant changes in ACTH and cortisol levels in both females and males after metformin treatment. Metformin use resulted in significant increase in luteinizing hormone (LH) and progesterone levels in males, while it was associated with significant increase in prolactin, follicular stimulating hormone (FSH), and dehydroepiandrostenedione-sulphate (DHEA-S) levels and significant decrease in total testosterone level in females. Conclusion Metformin treatment in females with prediabetes reduces BMI, waist circumference, fasting blood glucose, and HbA1c. The changes in the studied parameters differed significantly according to sex.

Impact of elevated serum estradiol/free testosterone ratio on male varicose veins in a prospective study.

Although some studies indicate an association between increased levels of estradiol and varicose veins in women, the role of sex hormones on varicose veins have a question mark in men. We investigated estradiol-2/free testosterone (E2/fT) ratio relationship on varicosity in 100 male patients dividing them into two groups. Group A (n = 46) had varicose veins with endocrinological problems, especially infertility, whereas group B (n = 54) also had varicose veins but no endocrinological problems. Venous blood samples were drawn from both groups in the morning to detect the levels of serum estradiol (E2), androstenedione, dehydroepiandrostenedione sulphate and free testosterone (fT). Patient history, physical examination, colour duplex ultrasound of both limbs and classification of CEAP were performed in both groups. Serum E2/fT ratio was calculated in correspondence with CEAP classification. CEAP classification is the varicosity classification, and it is concerned about the clinical class (C), etiology (E), anatomical distribution (A) and underlying pathophysiology (P). E2/fT ratio was significantly higher in group A (4.18 ± 0.54) compared with group B (2.98 ± 0.36). Moreover, there is a high correlation between serum E2/fT ratio and CEAP clinical classification in group A (4) compared with group B (2). Serum E2/fT ratio is associated with varicose veins in male patients.

Normal cut-off values for hyperandrogenaemia in Iranian women of reproductive age

ObjectiveThe normal cut-off values for hyperandrogenaemia differ between populations. Although there are various ways to determine the normal range of hormones in a population, use of a gold standard and the 95th percentile approach are not appropriate to diagnose hyperandrogenism. This study aimed to determine normative cut-off androgen levels for Iranian women of reproductive age using k-means cluster analysis. Study designThe subjects were 1126 women, aged 18–45 years, initially selected at random from women of reproductive age from different geographic regions of Iran. Serum levels of hormones were measured, and complete data were available for 923 women following the implementation of exclusion criteria. The reference group comprised 423 eumenorrhoeic non-hirsute women selected from the total population. Normative cut-off levels were computed using k-means cluster analysis (k=3). Analyses were also conducted based on age and body mass index (BMI) groups for the 923 subjects. ResultsCluster cut-off values for the total study population were similar to the 95th percentile values, whereas the percentiles for the free androgen index, total testosterone, dehydroepiandrostenedione sulphate and androstenedione were 79.3%, 74.1%, 96.7% and 71%, respectively, in the reference group. ConclusionsDetermination of cut-off values for hyperandrogenism based on age and BMI categories provides the most objective way of determining normal upper limits of androgen levels among Iranian women of reproductive age.

Androgen-secreting adrenocortical carcinoma

A 47-year-old woman presented with abdominal pain. She had recently noticed increased growth of facial hair and loss of scalp hair, and her menstrual cycle had become erratic. Abdominal examination revealed a right upperquadrant abdominal mass. A CT scan of the abdomen revealed a 10 cm adrenal mass (fi gure, A, B; green arrows) that was well encapsulated and heterogeneous. Endocrine studies were ordered to exclude a functioning adrenal neoplasm. The serum total testosterone concentration was 8·6 nmol/L (reference range 0·13–2·53 nmol/L) and the serum dehydroepiandrostenedione sulphate (DHEAS) con cen tration was 22 μmol/L (reference range 1·0–11·6 μmol/L). The free androgen index (ratio of testosterone to sex-hormone-binding globulin) was 30·2% (reference range 75% of tumour cells, and necrosis; fi gure, C [haematoxylin and eosin staining]). Immuno histochemistry was positive for melan-A, vimentin, synaptophysin, and epithelial membrane antigen. Cytoplasmic staining was positive with calretinin (fi gure, D) and inhibin (fi gure, E). The Ki-67 proliferative index was 32%. Mitotane was recommended as adjuvant therapy for 2 years. No evidence of recurrence or metastatic disease was recorded 12 months postoperatively and androgen concentrations remain suppressed. Women presenting with hirsutism or virilisation are more likely to have polycystic ovarian syndrome than an androgen-secreting adrenal neoplasm. Assessment of patients requires thorough hormonal investigation and imaging, including CT, MRI, and FDG-PET.

Breast size increment during pregnancy and breastfeeding in mothers with polycystic ovary syndrome: a follow‐up study of a randomised controlled trial on metformin versus placebo

To study the significance of breast size increment in pregnancy, and the impact of metformin during pregnancy on breastfeeding in women with polycystic ovary syndrome (PCOS). A follow-up study of a randomised controlled trial (the PregMet study). Eleven secondary care centres. Women with PCOS during pregnancy and postpartum. Women with PCOS were randomised to treatment with metformin or placebo from the first trimester to delivery. Questionnaires were sent to 240 participants 1 year postpartum: 186 responded. Pre-pregnancy and late-pregnancy brassiere size and breastfeeding patterns were registered, and androgen levels were measured in the mothers. No difference in breast size increment and breastfeeding were found between the placebo and metformin groups. Breast size increment correlated positively with the duration of both exclusive and partial breastfeeding, whereas body mass index (BMI) correlated negatively with the duration of partial breastfeeding. Dehydroepiandrostenedione-sulphate (DHEAS), testosterone and free testosterone index (FTI) in pregnancy did not correlate with breast size increment or duration of breastfeeding. Women with no change in breast size were more obese, had higher blood pressure, serum triglycerides and fasting insulin levels, and had a shorter duration of breastfeeding compared with those with breast size increment. Metformin and androgens had no impact on breastfeeding. Women with PCOS who had no breast size increment in pregnancy seem to be more metabolically disturbed and less able to breastfeed.

Relation between androgens and cardiovascular risk factors in a young population

The relationship between androgens and blood pressure, insulin resistance, lipid profile, adiponectin and hs-CRP in a young Middle-Eastern population has not been examined previously. We studied this relationship in a randomly selected population of Lebanese students. Three hundred and sixty-eight subjects (201 men and 167 women) aged 18-30 years were included in the study. Anthropometric and biological parameters [waist circumference (WC), systolic and diastolic blood pressure (SBP and DBP), triglycerides, total cholesterol, HDL cholesterol, homeostasis model assessment of insulin resistance (HOMA-IR), total testosterone (TT), dehydroepiandrostenedione sulphate (DHEAS), sex hormone-binding globulin (SHBG), adiponectin (ADP) and hs-CRP] were measured. In men, there were inverse associations of both TT and SHBG with body mass index (BMI), WC, HOMA-IR, triglycerides and hs-CRP. After adjustment for major confounders (BMI, WC, age and smoking), associations disappeared except for those between TT and hs-CRP, and for SHBG HOMA-IR, hs-CRP and triglycerides. In women, only SHBG was inversely associated with BMI, WC, HOMA-IR and hs-CRP and positively correlated with adiponectin. Except for the association between SHBG and adiponectin, these correlations disappeared after adjustment for confounders. Although DHEAS appeared to correlate negatively with blood pressure in men, this relationship disappeared after adjustment for confounders, while a relationship between DHEAS and triglycerides in women persisted after such adjustment. In multivariate regression analysis, SHBG was an independent predictor of hs-CRP, triglycerides and HOMA-IR in men and of adiponectin in women. Our results suggest that SHBG is independently associated with HOMA-IR, adiponectin, hs-CRP and triglycerides. A gender difference in these associations is observed. Further studies are needed to elucidate these findings.

Metformin 2,500 mg/day in the treatment of obese women with polycystic ovary syndrome and its effect on weight, hormones, and lipid profile

The objective of this study was to assess the efficacy and safety of metformin at the dosage of 2,500 mg/day in the treatment of obese women with PCOS and also to evaluate its effect on weight, hormones, and lipid profile. This study was a 4-month open-label clinical trial. Sixty-nine PCOS patients aged 20-35 were recruited in the study. Testosterone, free testosterone, sex hormone-binding globulin (SHBG), fasting insulin, dehydroepiandrostenedione-sulphate (DHEAS), FBS, LDH, HDL, TG, total cholesterol, body mass index (BMI), and waist-to-hip ratio were measured before treatment and after 4 months of treatment. Significant reductions in serum insulin, BMI, waist/hip ratio, and LDL were observed. In addition, a significant increase in SHBG was obtained. Over the 4 months of the trial, 12 patients faced nausea, six patients had bloating, five patients had diarrhea and two had headache; none of these symptoms were severe except for two cases that dropped out due to severe vomiting. The results of this study show that 2,500 mg daily dose of metformin in obese patients with PCOS is effective in the reduction of BMI, waist hip/ratio, LDL, serum insulin and increases SHBG. In general this dose was relatively safe and well tolerated.

High prevalence of polycystic ovary syndrome in women born small for gestational age

There is evidence that intrauterine growth restriction, resulting in newborn girls that are small for gestational age (SGA), may be related to the onset of polycystic ovary syndrome (PCOS). Thus, we studied whether women born SGA have a higher prevalence of PCOS than women born appropriate for gestational age (AGA). This was a prospective birth cohort study of 384 women born at term between June 1, 1978, and May 31, 1979, in Ribeirão Preto, Brazil. After exclusion, 165 women effectively participated in this study, of whom 43 were SGA and 122 were AGA. The prevalence of PCOS was analysed. At a mean age of 29 years, the women agreed to follow the study protocol, which included: anamnesis, physical examination, serum tests [follicle stimulating hormone, luteinizing hormone, total and free testosterone, dehydroepiandrostenedione sulphate, 17-OH-progesterone, fasting insulin, sex steroid-binding globulin (SHBG) and fasting glucose] and pelvic ultrasound. Data regarding gestational age, birthweight, age at menarche and maternal data were obtained from the files of the cohort. The adjusted relative risk (RR) values of the SGA, insulin resistance, body mass index, maternal smoking and parity variables were analysed using Poisson regression with robust adjustment of variance for the prediction of PCOS. The prevalence of PCOS was higher in the SGA group than in the AGA group [adjusted RR = 2.44, 95% CI (1.39-4.28)]. Hyperandrogenism was more prevalent in the SGA women than in the AGA women (P = 0.011). Circulating SHBG was lower in the SGA women than in the AGA women (P = 0.041), but fasting insulinemia was similar in both groups. The prevalence of PCOS in SGA women was twice as high as in AGA women in our study population.

Dehydroepiandrostenedione sulphate interferes in many direct immunoassays for testosterone

In response to a report that the Abbott Architect direct testosterone immunoassay was affected by dehydroepiandrostenedione sulphate (DHEAS) cross-reactivity, this study investigated the effect of DHEAS on other testosterone methods using a UK National External Quality Assessment Scheme (UK NEQAS) for steroid hormones special distribution. Separate male and female matrix pools of normal human serum were prepared and divided into three portions. Two portions from each matrix were spiked with 10.07 and 20.14. micromol/L DHEAS, respectively. Aliquots of these pools were distributed to participants in UK NEQAS for male and female testosterone at distribution 319 (July 2006). Median testosterone results for each method group were examined for an increase in testosterone concentration. In the female matrix, the effect on Abbott Architect was confirmed and shown also for Roche Elecsys, Roche E170 Modular and Beckman Access (includes some Beckman DxI users). In the male matrix, the effect on Abbott Architect, Roche Elecsys and Roche E170 Modular was similar in magnitude to that in the female matrix, but was smaller for Beckman Access/DxI. However, the effect on Immulite 2000 (includes some Immulite 2500 users) was greater in the male matrix. Indications of interference in other methods, used by only a few laboratories (notably Tosoh AIA and Wallac Delfia), were also observed. DHEAS interference in testosterone assays is a feature of many routine testosterone methods, and occurs in both male and female matrices.

Effect of spearmint (Mentha spicata Labiatae) teas on androgen levels in women with hirsutism

Mentha spicata Labiatae, known as spearmint and Mentha piperita Labiatae, known as peppermint can be used for various kinds of illnesses in herbal medicine and flavoring in industry. M. spicata Labiatae grows on the Anamas plateau of Yenithornarbademli town of Isparta, located in southwest part of Turkey. In this town, clinicians thought that consumption of tea steeped with M. spicata or M. piperita caused a diminished libido. Because antiandrogenic effects of spearmint and peppermint were found previously in rats, it was decided to observe the effect of this herbal tea on the androgen levels in hirsute women.Twenty-one female hirsute patients, 12 with polycystic ovary syndrome and 9 with idiopathic hirsutism were included to the study. They were took a cup of herbal tea which was steeped with M. spicata for 5 days twice a day in the follicular phase of their menstrual cycles. After treatment with spearmint teas, there was a significant decrease in free testosterone and increase in luteinizing hormone, follicle-stimulating hormone and estradiol. There were no significant decreases in total testosterone or dehydroepiandrostenedione sulphate levels. Spearmint can be an alternative to antiandrogenic treatment for mild hirsutism. Further studies are needed to test the reliability of these results and the availability of spearmint as a drug for hirsutism.

Liquorice in Moderate Doses Does Not Affect Sex Steroid Hormones of Biological Importance although the Effect Differs between the Genders

Background/Aim: Liquorice is commonly consumed, at least in the western world, and we have earlier shown that even moderate doses of liquorice have significant effects on the cortisol metabolism by inhibiting 11β-hydroxysteroid dehydrogenase type 2. The suggestion that liquorice decreases the testosterone levels in men makes it vital to study the effect of moderate doses of liquorice on sex steroid hormones. Methods: Fifteen women and 21 men (healthy volunteers and subjects with essential hypertension) consumed 100 g of liquorice (150 mg glycyrrhetinic acid) daily in a 9-week, open-treatment trial. Blood and 24-hour urine samples were collected for hormone analysis before and after 4 weeks of liquorice consumption and 4 weeks after cessation of liquorice intake. Results: The liquorice induced a moderate decrease in the serum concentrations of dehydroepiandrostenedione sulphate in men (p = 0.002). The relative change in serum levels of dehydroepiandrosterone sulphate differed between the genders (p = 0.03). No significant changes were observed in the serum testosterone levels after 4 weeks of liquorice consumption, and the urine excretion of androgens (etiocholanolone and androstenedione) did not change. Conclusions: Liquorice in moderate doses primarily affects the cortisol metabolism and only marginally the androgen hormones. Gender may influence the action of liquorice.

Serum uric acid, dehydroepiandrosterone sulphate, and apolipoprotein E genotype in benign vs. progressive multiple sclerosis

The majority of patients with multiple sclerosis (MS) experience gradual progression of disability, either as secondary progressive MS (SPMS) or primary progressive MS (PPMS). A subgroup with relapsing-remitting MS shows a benign course with little or no disease progression and minimal disability decades after the first manifestations, so called benign MS (BMS). In our search to identify factors that are associated with progression of MS, we investigated serum levels of uric acid and dehydroepiandrostenedione sulphate (DHEAS), and apolipoprotein (apo)E genotype in 28 patients with BMS, 33 with SPMS, 21 with PPMS, and 29 healthy individuals. We found no significant changes in uric acid levels and apoE genotype between the four groups. Mean DHEAS levels were lower in MS patients compared with healthy controls (P = 0.049), but there were no significant differences between the clinical subgroups of MS. In patients with SPMS and PPMS there was no correlation between progression rate and serum levels of either uric acid or DHEAS. Our results suggest that serum levels of uric acid and DHEAS, and apoE genotype do not differ between patients with a benign and progressive course of MS.

Growth and adrenal androgen status at 7 years in very low birth weight survivors with and without bronchopulmonary dysplasia

Aims: To evaluate whether 7 year old VLBW (very low birth weight, <1500 g) survivors with and without bronchopulmonary dysplasia (BPD) evince similar growth status and higher adrenal androgen (AA)...

Serum adrenal androgens in women with primary operable breast cancer and their relationship with age and body mass index

Several studies have found elevated levels of adrenal androgens in postmenopausal women and depressed levels in premenopausal women with breast cancer, suggesting a role for adrenal androgens in the aetiology of breast cancer. We have measured serum dehydroepiandrosterone sulphate and androstenedione in 81 women with primary operable breast cancer and 62 age-matched controls. Results showed that serum levels of both adrenal androgens fell significantly with age in women with breast cancer ( P=0.003). However, no relationship was observed between serum adrenal androgen levels and body mass index in either women with breast cancer or controls. Dehydroepiandrosterone sulphate levels were elevated in postmenopausal women with breast cancer compared to controls, and this was not due to preoperative stress. No differences were observed in androstenedione levels between premenopausal or postmenopausal women with breast cancer and controls, nor were dehydroepiandrostenedione sulphate levels significantly different between premenopausal women with breast cancer and controls. These results suggest that dehydroepiandrosterone sulphate has a role in the aetiology of postmenopausal breast cancer.

The role of androgens in the aetiology and pathology of pre-eclampsia

Pre-eclampsia and eclampsia, along with its complications such as prematurity, intrauterine growth retardation, perinatal asphyxia and abruptio placenta, continues to be one of the major causes of maternal and fetal morbidity and mortality. Due to its morbid course, it is necessary to identify those at risk for the illness and take precautions. However, the lack of knowledge about the aetiology makes it difficult to assess risk factors. We studied the role of the serum androgens in the pathogenesis of pre-eclampsia. Ninety severe pre-eclamptic, 10 mild pre-eclamptic and 50 normotensive pregnant women were studied. The patient and control groups' blood total testosterone, free oestriol (FE3), FAI (free androgene index), sex hormone binding globulin (SHBG) and dehydroepiandrostenedione sulphate (DHEA-S) values were compared. The sex, weight, APGAR scores and hospitalisation in the neonatal intensive care unit of the babies were also evaluated. No statistically significant difference in the total testosterone and free oestriol values was found between the groups. SHBG was found to be higher in the pre-eclamptic group, whereas free testosterone and DHEA-S were higher in the normotensive group. Compared to the severe pre-eclamptic group, no difference was found in the total testosterone levels, whereas free testosterone levels were significantly higher in the mild pre-eclamptic group. We reached the opinion that androgens do not play a clinically significant role in the pathogenesis of pre-eclampsia.

Clinical, ultrasound and hormonal markers of androgenicity in acne vulgaris

Androgenic stimulation of sebaceous glands is an important factor in the development of acne. We examined 36 females (aged 14-34 years), selected because none had received oral contraceptives, anti-androgen therapy, or systemic antibiotics during the previous year, or isotretinoin therapy, prior to their participation in the study. Subjects were divided into groups on the basis of acne severity, as follows: physiological, mild and moderate. Only two patients had polycystic ovaries on ultrasound examination. Seven patients had irregular menses; none had evidence of hirsutism. We found that the severity of acne, based on the acne grade, was highly correlated with the inflammatory lesion count, and less correlated with the sebum excretion rate. Either acne grade or inflammatory lesion count could be related to some of the five androgenic hormone determinants; free testosterone (TESTOS), delta 4 androstenedione (DELTA 4), sex hormone binding globulin (SHBG), dehydroepiandrostenedione sulphate (DHEAS) and dihydrotestosterone (DHT). Multiple linear regression analysis determined the best model for predicting ACNE scores as involving DELTA 4 and DHEAS (positive effects), and SHBG (negative effect), P < 0.005, R2 = 0.36). In none of the patients were the levels of DHEAS or SHBG outside the normal range. The findings in the two patients with polycystic ovaries did not differ significantly from those in the remainder of the patients.

Effects of human pituitary extracts on androgen biosynthesis by human adrenals in vitro.

SUMMARY The effects of human pituitary extracts upon steroid biosynthesis by pieces of human adrenal tissue incubated in Krebs—Ringer bicarbonate buffer have been studied. Experiments with both adult and foetal adrenal tissue have been carried out. Radioactive precursor (either [7-3H]pregnenolone or [4-14C] progesterone) was added to each incubation vessel and the synthesis of radioactively labelled products was estimated by reverse isotope dilution. Individual steroids were characterized by paper and thin-layer chromatography; the radiochemical purity of cortisol, androstenedione, dehydroepiandrostenedione (DHA) and DHA sulphate was established following derivative formation. There was no consistent effect of the pituitary preparations upon the biosynthesis of cortisol, DHA or DHA sulphate, nor upon the ratio of cortisol to the combined production of DHA and DHA sulphate. However, the production of androstenedione from both [7-3H]pregnenolone and [4-14C]progesterone was significantly increased in the presence of pituitary and gonadotrophic preparations.