9α-hydroxy-4-androstene-3,17-dione (9-OH-AD) is an important intermediate in the steroidal drugs production. 3-ketosteroid-9α-hydroxylase (KSH), a two protein system of KshA and KshB, is a key-enzyme in the microbial steroid ring B-opening pathway. KSH catalyzes the transformation of 4-androstene-3,17-dione (AD) into 9-OH-AD specifically. In the present study, the putative KshA and KshB genes were cloned from Mycobacterium smegmatis mc(2)155 and Gordonia neofelifaecis NRRL B-59395 respectively, and were inserted into the expression vector pNIT, the co-expression plasmids of kshA-kshB were obtained and electroporated into Mycobacterium sp. NRRL B-3805 cells. The recombinants were used to transform steroids, the main product was characterized as 9α-hydroxy-4-androstene-3,17-dione (9-OH-AD), showing that kshA and kshB were expressed successfully. Different from the original strain Mycobacterium sp. NRRL B-3805 that accumulates 4-androstene-3,17-dione, the recombinants accumulates 9α-hydroxy-4-androstene-3,17-dione as the main product. This results indicates that the putative genes kshA, kshB encode active KshA and KshB, respectively. The process of biotransformation was investigated and the results show that phytosterol is the most suitable substrate for biotransformation, kshA and kshB from M. smegmatis mc(2)155 seemed to exhibit high activity, because the resultant recombinant of them catalyzed the biotransformation of phytosterol to 9-OH-AD in a percent conversion of 90%, which was much higher than that of G. neofelifaecis NRRL B-59395. This study on the manipulation of the ksh genes in Mycobacterium sp. NRRL B-3805 provides a new pathway for producing steroid medicines.
Androstenedione ; analogs & derivatives ; biosynthesis ; Bacterial Proteins ; genetics ; metabolism ; Biotransformation ; Ketosteroids ; Mixed Function Oxygenases ; genetics ; metabolism ; Mycobacterium ; metabolism ; Mycobacterium smegmatis ; enzymology ; Plasmids

Congenital adrenal hyperplasia refers to a group of autosomal recessive disorders that is defective in the synthesis of cortisol. The enzymes most often affected are 21-hydroxylase and 11beta hydroxylase. The low levels of cortisol stimulate the pituitary gland to release ACTH. Chronic elevation of the ACTH level causes bilateral adrenal hyperplasia and a secondary increase in androgen formation. We examined a 19 year-old woman presented with clitoral hypertrophy and vaginal spotting. The subjects basal level of serum cortisol was low, but the serum levels of ACTH, 17a-hydroxyprogesterone, deoxy-corticosterone were elevated. The urinary excretions of 17-ketosteroids and 17-hydroxycorticosteroids were also increased. The karyotyping study and transrectal ultrasonography showed normal findings. The patient underwent clitoris reduction surgery and received hydrocortisone. To the best of our knowledge, this is the first case of 11beta-Hydroxylase deficiency in Korea.
17-Hydroxycorticosteroids ; 17-Ketosteroids ; Adrenal Hyperplasia, Congenital* ; Adrenocorticotropic Hormone ; Clitoris ; Female ; Humans ; Hydrocortisone ; Hyperplasia ; Hypertrophy ; Karyotyping ; Korea ; Metrorrhagia ; Pituitary Gland ; Steroid 21-Hydroxylase ; Ultrasonography ; Young Adult

Bilateral adrenal tuberculosis is a rare disease and often occurs bilaterally. We report a case of bilateral adrenal masses due to tuberculosis with adrenal insufficiency. The patient was a 39- year-old man who had complained of intermittent pain of right upper quadrant and general weakness. The plasma levels of cortisol and catecholamine were normal. The levels of 24-hour urinary catecholamine and VMA were also normal. But the levels of 24-hour urinary 17- hydroxycorticosteroids and 17-ketosteroids were reduced. Abdominal CT showed about 6.5 x4.8 x 5.4cm sized left adrenal mass and 4.0 x 2.8 x 3.6cm sized right adrenal mass with calcification. The result of sono-guided percutaneous needle biopsy was adrenal tuberculosis. The patient was treated with antituberculous chemotherapy and hormonal replacement. But the masses are unchanged during 5-months follow-up.
17-Ketosteroids ; Adrenal Glands ; Adrenal Insufficiency ; Biopsy, Needle ; Drug Therapy ; Follow-Up Studies ; Humans ; Hydrocortisone ; Hydroxycorticosteroids ; Plasma ; Rare Diseases ; Tomography, X-Ray Computed ; Tuberculosis*

PURPOSE: Adrenocortical tumors are uncommon in children and comprise only a small proportion of primary adrenal neoplasms. The biologic behavior of these tumors may be very difficult to predict, and their rarity has hindered identification of clinical characteristics. Patients with functioning tumors have excessive steroid hormone production, and the clinical manifestation depends on the predominant hormone produced. The detection of nonfunctioning tumors is not easy and the diagnosis may be delayed. Benign tumors can be cured by complete surgical excision, but malignant cases have poor response to treatment and worse prognosis. Early diagnosis and proper management are very important because of the large proportion of functioning malignant tumors in children. We report clinical features of adrenocortical tumors in children that may be of help in the early detection, proper management, and assessment of prognosis of patients. METHODS: We reviewed the clinical characteristics of 14 cases of adrenocortical tumors, among 85 children diagnosed with adrenal tumors, who visited the Severance Hospital, College of Medicine, Yonsei University, from January 1970 to July 1996. RESULTS: 85 Patients were diagnosed with adrenal tumors. Among them, 71 cases 83.5%) were tumors of the adrenal medulla, neuroblastoma and pheochromocytoma, and 14 cases (16.5%) were adrenocortical tumors, consisting of 5 cases of adenoma, 7 cases of carcinoma, and 2 cases unspecified. The age distribution ranged from 16 months to 14 years of age, and the mean was 5 years & 11 months (median 4 years & 2 months). Sex distribution revealed a male to female ratio of 1:1.33. The left to right ratio was 3.7:1, showing a left side predominance. 13 Cases (92.9%) were functioning tumors: 12 cases (92.3%) had clinical evidence of androgen excess, among which 6 cases (46.2%) were associated with Cushing's syndrome, and 1 case was compatible with primary aldosteronism. Serum cortisol, urinary 17-ketosteroids and 17-hydroxycorticosteroids concentrations were measured in 11 cases and urinary concentrations of 17-ketosteroids were elevated in all 11 cases (100%), while 17-hydroxycorticosteroids were elevated in 4 cases (36.4%). Abnormalities of serum cortisol were found in all cases except 1: serum cortisol concentrations were abnormally elevated in 5 cases (45.5%), and the remainder (5 cases, 45.5%) showed loss of diurnal variation. Dexamethasone suppression test was performed in 9 cases, and all (100%) showed no suppression. Preoperative radiologic studies included abdominal sonograms, CT or MRI scans, and angiography. Histology showed carcinomas to be bigger and heavier than adenomas, and microscopically carcinomas had necrosis, calcifications, and invasions of vessels. Distant metastases were found in 4 cases (12.7%). Adrenalectomy with complete surgical excision was performed in 12 cases. Long-term follow-up was possible in 10 patients after operation: 3 patients initially diagnosed with adenoma survived without tumor recurrence over a year, and among 6 carcinoma patients, 4 expired within a year, and 2 survived, with one patient currently undergoing postoperative chemotherapy. Of the 10 patients currently under going follow-up, one patient was initially diagnosed with a histologically unspecified tumor, and has survived 4 years after operation. CONCLUSIONS: When adrenocortical neoplasms are suspected by clinical symptoms and laboratory findings, abdominal ultrasonogram, CT or MRI scans must be performed immediately. Early detection and proper management are important for better prognosis, but are often delayed in the majority of cases. Ultimately, pediatricians need to be familiar with clinical characteristics and laboratory findings of adrenocortical tumors, bearing in mind the possibility of diagnosis in children.
17-Hydroxycorticosteroids ; 17-Ketosteroids ; Adenoma ; Adrenal Gland Neoplasms ; Adrenal Medulla ; Adrenalectomy ; Age Distribution ; Angiography ; Child ; Cushing Syndrome ; Dexamethasone ; Diagnosis ; Drug Therapy ; Early Diagnosis ; Female ; Follow-Up Studies ; Humans ; Hydrocortisone ; Hyperaldosteronism ; Magnetic Resonance Imaging ; Male ; Necrosis ; Neoplasm Metastasis ; Neuroblastoma ; Pheochromocytoma ; Prognosis ; Recurrence ; Sex Distribution ; Ultrasonography

Patients undergoing thoracotomy experience severe postoperative pain and marked respiratory impairment. Analgesics(narcotics or loeal anesthetics) administered via epidural catheter in epidural space have been shown to provide postoperative analgesia and improve respiratory mechanics after thoracotomy. Several different methods have been utilized in an attempt to reduce pain and pulmonary mechanics after thoracotomy. These include epidural blocks using local anesthetics, epidural narcotics, ketamine, steroid, and clonidine. These methods have been shown to provide pain relief with relative preservation of lung volumes in the postoperative period, but have disadvantages. Especially epidural local anesthetics may cause hypotension and motor blockade of lower extremities, and epidural narcotics may cause pruritus, nausea and vomiting, urinary retension and respiratory depression. In an attempt to provide excellent analgesia and improve pulmonary mechanics after thoracotomy and to decrease the side effects associated with the intermittent bolus administration of epidural narcotics or local anesthetics, we performed a study of continuously administered epidural infusion of small concentration of fentanyl combined with low concentration of bupivacaine. Twenty eight patients undergoing thoracotomy were randomized into groups based upon a postoperative pain regimen as indicated: Group I: intermittent intramuscualr injection of nalbuphine 0.2 mg/kg(n=13), Group II: continuous epidural injection of mixtures of 0.2/ bupivacaine and fentanyl 3 ug/ml(n= 15). Two, 8, 24 and 48 hours postoperative, the following indices were measured: visual analogue pain scale, vital capacity, tidal volume, arterial blood gas analysis(pH, PaCo2, PaO2), side effects, and 24 hour urine 17-ketosteroids. The results were as follows: 1) Pain score was evaluated by visual analogue pain scale postoperatively and the pain scores significantly decresed in group II as compaired with those in group L 2) Vital capacity and tidal volume in group II were more improved than group I. 3) There was no difference in arterial blood gas analysis except for decreased PaO2 at 2 hour and 24 hour compared with preoperative value in group L 4) Major complications in group II were two cases of nausea and vomiting, one case of urinary retension, whereas only I patient in group I complained of nausea and vomiting. 5) No significant difference occurred in 24 hour urine l7-ketosteroid at 24 hour and 48 hour postoperatively in group I and group II, which were within normal limits.
17-Ketosteroids ; Analgesia* ; Anesthetics, Local ; Blood Gas Analysis ; Bupivacaine* ; Catheters ; Clonidine ; Epidural Space ; Fentanyl* ; Humans ; Hypotension ; Injections, Epidural ; Ketamine ; Lower Extremity ; Lung ; Mechanics ; Nalbuphine ; Narcotics ; Nausea ; Pain Measurement ; Pain, Postoperative ; Postoperative Period ; Pruritus ; Respiratory Insufficiency ; Respiratory Mechanics ; Thoracotomy* ; Tidal Volume ; Vital Capacity ; Vomiting

AIMTo establish a method to determine the isotope ratios of 13C to 12C of dehydroepiandrosterone and its metabolites in urine, for detecting the source of dehydroepiandrosterone or its metabolites.

METHODSPreliminary separation of endogenous anabolic androgenic steroids could be achieved using solid phase extraction, enzymolysis and thin layer chromatography. The source of dehydroepiandrosterone and other endogenous anabolic androgenic steroids could be detected by their delta values with gas chromat ography-combustion-isotope ratio mass spectrometry.

RESULTSThe 5 values of some metabolites of dehydroepiandrosterone reduced after the administration of dehydroepiandrosterone preparation. In these cases the data indicated that exogenous anabolic androgenic steroids were administrated.

CONCLUSIONThe source of dehydroepiandrosterone or its metabolites in urine could be detected by measuring their delta values with this method.

Adult ; Androstane-3,17-diol ; urine ; Androsterone ; urine ; Chromatography, Thin Layer ; methods ; Dehydroepiandrosterone ; metabolism ; Doping in Sports ; Etiocholanolone ; urine ; Female ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Male ; Pregnanetriol ; urine ; Substance Abuse Detection ; methods

BACKGROUND: Hyperprolactinemia has been linked with hyperandrogenism and hirsutism in some women. High plasma Dihydroandrosterone and DHA-S levels were reported in patients with hyperprolactinemia and a dissociation of adrenal androgen and cortisol secretion occurs in normal subjects. The mechanism has not been elucidated, but it has been suggested that pituitary factors other than ACTH modulate adrenal androgen synthesis, One candidate hormone is prolactin. Adrenal tissue has been found to possess prolactin receptors and prolactin has been shown to act synergistically with ACTH and lowers the activity of the enzyme 5a-reductase or 3B-hydroxysteroid dehydrogenase (3B-HSD). The aim of this study was to investigate the secretion of adrenal androgen metabolites in patients with idiopathic hyperprolactinemia and prolactinoma and to deterrnine the relationship with prolactin and androgens. METHODS: We measured 24 hour-urinary DHEA, androstenedione, androsterone, pregnenolone, tetrahydrocorticoid and cortisol in 16 normal controls and 5 patients with idiopathic hyperprolac-tinemia (HP) and 12 patients with prolactonoma in the early follicular phase. RESULTS: Urinary DHEA, AD (androsteredione), and androsterone, the metabolites of adrenal androgen, were significantly higher in both patients with idiopathic HP and prolactinoma compared with those in normal controls (p<0.05), whereas they were not different in both disease groups. Urinary pregnenolone levels, early metabolite of adrenal steroid synthesis, were lower in patients. In contrast, urinary tetrahydorcortisol and cortisol were higher in patients compared to controls. There was no difference in DHEA:androsterone ratio between patients and controls. And there were no correlation between prolactin levels and the levels of androgenic metabolites or clinical symptoms. CONCLUSION: Prolactin has a tropic effct on the secretion of androgens and steroids by the adrenal cortex. But prolactin levels were not correlated with androgen levels or clinical symptoms (amenorrhea), and it might have little effect on lowering the activity of 3B-HSD.
Adrenal Cortex ; Adrenocorticotropic Hormone ; Androgens ; Androstenedione ; Androsterone ; Dehydroepiandrosterone ; Female ; Follicular Phase ; Hirsutism ; Humans ; Hydrocortisone ; Hyperandrogenism ; Hyperprolactinemia* ; Oxidoreductases ; Plasma ; Pregnenolone ; Prolactin ; Prolactinoma ; Receptors, Prolactin ; Steroids

OBJECTIVE: The aim of this study was to measure maternal plasma androgens, estrogen and leptin levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 32 healthy pregnant women as well as 28 pregnant women with severe preeclampsia. Plasma leptin, total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea (p<0.05). In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups. The plasma levels of leptin were not significantly increased in the preeclampsia group. Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens ; Female ; Gestational Age ; Gonadal Steroid Hormones* ; Hematocrit ; Humans ; Leptin* ; Parity ; Plasma* ; Pre-Eclampsia* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid

OBJECTIVE: The aim of this study was to measure maternal plasma androgens, estrogen and leptin levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 32 healthy pregnant women as well as 28 pregnant women with severe preeclampsia. Plasma leptin, total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea (p<0.05). In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups. The plasma levels of leptin were not significantly increased in the preeclampsia group. Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens ; Female ; Gestational Age ; Gonadal Steroid Hormones* ; Hematocrit ; Humans ; Leptin* ; Parity ; Plasma* ; Pre-Eclampsia* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid

OBJECTIVE: The aim of this study was to measure maternal plasma androgens and estrogen levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 28 healthy pregnant women as well as 24 pregnant women with severe preeclampsia. Plasma total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea. In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups (p>0.05). Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens* ; Female ; Gestational Age ; Hematocrit ; Humans ; Parity ; Plasma* ; Pre-Eclampsia ; Pregnancy ; Pregnancy Trimester, Third* ; Pregnancy* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid