Congenital adrenal 11-β hydroxylase deficiency is a rare autosomal recessive syndrome characterized by deficient cortisol synthesis and testicular masses. It is extremely difficult to distinguish testicular tumors caused by this syndrome from Leydig cell tumors. As management for each differs, it is important to differentiate the syndromes from each other. Hereby, we present the case of two brothers affected by 11-β hydroxylase deficiency and presenting with bilateral testicular masses. Two differential diagnoses were noticed for both patients: testicular adrenal rest tumors (TART) and Leydig cell tumor (LCT). In this study the tumors were yellow, firm, and non-tender with intra-testicular location. Histological studies showed cells in a cluster arrangement with low lipochrome pigment concentration. Tumors were unresponsive to ACTH suppression therapy, but a drop in levels of plasma testosterone and urinary <em>17</em>-<em>ketosteroids</em> occurred after surgical treatment. Considering all above, they were finally diagnosed as having Leydig cell tumors. Both cases were managed by testis-sparing surgery.

The human aldo-keto reductase (AKR) 1C3, also known as type-5 <em>17</em>β-hydroxysteroid dehydrogenase and prostaglandin F synthase, has been suggested as a therapeutic target in the treatment of prostate and breast cancers. In this study, AKR1C3 inhibition was examined by Brazilian propolis-derived cinnamic acid derivatives that show potential antitumor activity, and it was found that baccharin (1) is a potent competitive inhibitor (K(i) 56 nM) with high selectivity, showing no significant inhibition toward other AKR1C isoforms (AKR1C1, AKR1C2, and AKR1C4). Molecular docking and site-directed mutagenesis studies suggested that the nonconserved residues Ser118, Met120, and Phe311 in AKR1C3 are important for determining the inhibitory potency and selectivity of 1. The AKR1C3-mediated metabolism of <em>17</em>-<em>ketosteroid</em> and farnesal in cancer cells was inhibited by 1, which was effective from 0.2 μM with an IC(50) value of about 30 μM. Additionally, 1 suppressed the proliferation of PC3 prostatic cancer cells stimulated by AKR1C3 overexpression. This study is the first demonstration that 1 is a highly selective inhibitor of AKR1C3.

KSI (<em>ketosteroid</em> isomerase) from Comamonas testosteroni is a homodimeric enzyme that catalyses the allylic isomerization of Delta5-3-<em>ketosteroids</em> to their conjugated Delta4-isomers at a reaction rate equivalent to the diffusion-controlled limit. Based on the structural analysis of KSI at a high resolution, the conserved cis-Pro39 residue was proposed to be involved in the proper positioning of Asp38, a critical catalytic residue, since the residue was found not only to be structurally associated with Asp38, but also to confer a structural rigidity on the local active-site geometry consisting of Asp38, Pro39, Val40, Gly41 and Ser42 at the flexible loop between b-strands B1 and B2. In order to investigate the structural role of the conserved cis-Pro39 residue near the active site of KSI, Pro39 was replaced with alanine or glycine. The free energy of activation for the P39A and P39G mutants increased by 10.5 and 16.7 kJ/mol (2.5 and 4.0 kcal/mol) respectively, while DG(U)H2O (the free-energy change for unfolding in the absence of urea at 25.00+/-0.02 degrees C) decreased by 31.0 and 35.6 kJ/mol (7.4 and 8.5 kcal/mol) respectively, compared with the wild-type enzyme. The crystal structure of the P39A mutant in complex with d-equilenin [d-1,3,5(10),6,8-estrapentaen-3-ol-<em>17</em>-one], a reaction intermediate analogue, determined at 2.3 A (0.23 nm) resolution revealed that the P39A mutation significantly disrupted the proper orientations of both d-equilenin and Asp38, as well as the local active-site geometry near Asp38, which resulted in substantial decreases in the activity and stability of KSI. Upon binding 1-anilinonaphthalene-8-sulphonic acid, the fluorescence intensities of the P39A and P39G mutants were increased drastically, with maximum wavelengths blue-shifted upon binding, indicating that the mutations might alter the hydrophobic active site of KSI. Taken together, our results demonstrate that the conserved cis-Pro39 residue plays a crucial role in the proper positioning of the critical catalytic base Asp38 and in the structural integrity of the active site in KSI.

Analyses of 24-hour specimens of urine from healthy adult males for androsterone and etiocholanolone produced values which, when calculated as discriminant scores, discriminated between heterosexual and exclusively homosexual individuals. This confirms a previous study.No significant differences were found between heterosexuals and homosexuals in parental ages, secondary sex characteristics, genitalia, anthropometry, <em>17</em>-<em>ketosteroids</em>, and <em>17</em>-ketogenic steroids.A significant difference was found between the heterosexual group and homosexual group in the number of homosexual relatives in the immediate and extended families.

Actinobacteria comprise diverse groups of bacteria capable of full degradation, or modification of different steroid compounds. Steroid catabolism has been characterized best for the representatives of suborder Corynebacterineae, such as Mycobacteria, Rhodococcus and Gordonia, with high content of mycolic acids in the cell envelope, while it is poorly understood for other steroid-transforming actinobacteria, such as representatives of Nocardioides genus belonging to suborder Propionibacterineae. Nocardioides simplex VKM Ac-2033D is an important biotechnological strain which is known for its ability to introduce ∆(1)-double bond in various 1(2)-saturated 3-<em>ketosteroids</em>, and perform convertion of 3β-hydroxy-5-ene steroids to 3-oxo-4-ene steroids, hydrolysis of acetylated steroids, reduction of carbonyl groups at C-<em>17</em> and C-20 of androstanes and pregnanes, respectively. The strain is also capable of utilizing cholesterol and phytosterol as carbon and energy sources. In this study, a comprehensive bioinformatics genome-wide screening was carried out to predict genes related to steroid metabolism in this organism, their clustering and possible regulation. The predicted operon structure and number of candidate gene copies paralogs have been estimated. Binding sites of steroid catabolism regulators KstR and KstR2 specified for N. simplex VKM Ac-2033D have been calculated de novo. Most of the candidate genes grouped within three main clusters, one of the predicted clusters having no analogs in other actinobacteria studied so far. The results offer a base for further functional studies, expand the understanding of steroid catabolism by actinobacteria, and will contribute to modifying of metabolic pathways in order to generate effective biocatalysts capable of producing valuable bioactive steroids.

A built-in (genetically determined) about-7-day (circaseptan) period comes to the fore as a desynchronized feature of human time structure in the urinary excretion of <em>17</em>-<em>ketosteroids</em> by a clinical healthy man: during several years following an endocrine intervention (the self-administration of testosterone suppositories), a circaseptan rhythm (which during the preceding decade had revealed a period of precisely 7 days) deviated slightly, yet with statistical significance, from the environmental week. A second line of evidence for an intrinsic circaseptan component stems from the demonstration of statistically significant differences in timing of a circaseptan rhythm in springtail oviposition. A third line of evidence documents prominent circaseptan rhythmicity after the application of a single stimulus (devoid in itself of any circaseptan information). Such single stimulus induction, amplification and/or synchronization also documents the clinical and biologic importance of built-in circaseptan rhythms that were previously often misinterpreted as being purely reactive: a circaseptan spectral component is remarkably prominent in mammalian organ transplant rejection, both in the clinic and in the laboratory. In the latter case, in the absence of any weekly cycles in hospital routine, including treatment schedules, circaseptan components characterize the rejection of the rat kidney, pancreas and heart. Much additional information here reviewed reveals the occurrence of periods of about 7 days. Their implications for transplant and other chronoimmunology as well as biology in general, and their clinical applications in drug treatment, include the need to weld circaseptan timing to circadian timing and dosing. A dramatic documentation of this need stems from the circumstance that pretreatment for one week with the same total dose of the same substance (a polysaccharide - Lentinan) accelerates or retards cancerous growth (hence shortens or lengthens survival) as a function of interactive circaseptan and circadian rhythms.

3 alpha-Hydroxysteroid dehydrogenases (3 alpha-HSDs) catalyze the conversion of 3-<em>ketosteroids</em> to 3 alpha-hydroxy compounds. The best known 3 alpha-HSD activity is the transformation of the most potent natural androgen, dihydrotestosterone, into 5 alpha-androstan-3 alpha,<em>17</em> beta-diol (3 alpha-diol), a compound having much lower activity. Previous reports show that 3 alpha-HSDs are involved in the metabolism of glucocorticoids, progestins, prostaglandins, bile acid precursors, and xenobiotics. 3 alpha-HSDs could, thus, play a crucial role in the control of a series of active steroid levels in target tissues. In the human, type 1 3 alpha-HSD was first identified as human chlordecone reductase. Recently, we have isolated and characterized type 3 3 alpha-HSD that shares 81.7% identity with human type 1 3 alpha-HSD. The transfection of vectors expressing types 1 and 3 3 alpha-HSD in transformed human embryonic kidney (HEK-293) cells indicates that both enzymes efficiently catalyze the transformation of dihydrotestosterone into 3 alpha-diol in intact cells. However, when the cells are broken, the activity of type 3 3 alpha-HSD is rapidly lost, whereas the type 1 3 alpha-HSD activity remains stable. We have previously found that human type 5 <em>17</em> beta-HSD which possesses 84% and 86% identity with types 1 and 3 3 alpha-HSD, respectively, is also labile, whereas rodent enzymes such as mouse type 5 <em>17</em> beta-HSD and rat 3 alpha-HSD are stable after homogenization of the cells. The variable stability of different enzymatic activities in broken cell preparations renders the comparison of different enzymes difficult. RNA expression analysis indicates that human type 1 3 alpha-HSD is expressed exclusively in the liver, whereas type 3 is more widely expressed and is found in the liver, adrenal, testis, brain, prostate, and HaCaT keratinocytes. Based on enzymatic characteristics and sequence homology, it is suggested that type 1 3 alpha-HSD is an ortholog of rat 3 alpha-HSD while type 3 3 alpha-HSD, which must have diverged recently, seems unique to human and is probably more involved in intracrine activity.

Aldo-Keto-Reductase 1C3 (type 5 <em>17</em>β-hydroxysteroid dehydrogenase (HSD)/prostaglandin (PG) F2α synthase) is the only <em>17</em>β-HSD that is not a short-chain dehydrogenase/reductase. By acting as a <em>17</em>-<em>ketosteroid</em> reductase, AKR1C3 produces potent androgens in peripheral tissues which activate the androgen receptor (AR) or act as substrates for aromatase. AKR1C3 is implicated in the production of androgens in castration-resistant prostate cancer (CRPC) and polycystic ovarian syndrome; and is implicated in the production of aromatase substrates in breast cancer. By acting as an 11-ketoprostaglandin reductase, AKR1C3 generates 11β-PGF2α to activate the FP receptor and deprives peroxisome proliferator activator receptorγ of its putative PGJ2 ligands. These growth stimulatory signals implicate AKR1C3 in non-hormonal dependent malignancies e.g. acute myeloid leukemia (AML). AKR1C3 moonlights by acting as a co-activator of the AR and stabilizes ubiquitin ligases. AKR1C3 inhibitors have been used clinically for CRPC and AML and can be used to probe its pluripotency.

In 167 women, ages 41 +/- 3.5 years, who constituted 80 per cent of nuns of this age group in the Omaha area, body dimensions and radiogrammetric indices were determined in (1) radius; (2) femoral shaft; (3) second metacarpal. Calcium, phosphorus and nitrogen balance were measured and calcium kinetics were calculated following oral and intravenous administration of two radioisotopes of calcium. The bone indices were significantly correlated to each other. The femoral diameter was correlated to all the kinetic variables, the radial to many, and the metacarpal to a few. All bone diameters were correlated to height, but only the femoral diameter to body weight. Femoral shaft diameter increased with time after the menopause, but was not correlated to age. There was no correlation between any bone variable and dietary calcium, absorbed calcium, or calcium balance. High dietary calcium was associated with lower bone resorption. There was no correlation between any bone variable and the urinary excretion of estrogens or <em>17</em> <em>ketosteroids</em> or <em>17</em> hydroxycorticosteroids. Although different bones of the skleton are qualitatively and quantitatively related, in survey work the femoral shaft should be included as a skeletal marker in addition to the second metacarpal. Femoral expansion occurs in women after the menopause. In normal women, dietary calcium is unrelated to skeletal indices.

Oral testosterone undecanoate (TU) in arachis oil has been evaluated with a view to its possible use as a means of androgen replacement therapy. A single 100 mg dose was found to elevate plasma androgen levels and urinary <em>17</em>-<em>ketosteroid</em> excretion in 6 normal men. Ninety mg/day and 60 mg/day doses taken by a hypogonadal man resulted in sustained levels of androgen which appeared physiological when measured by radioimmunoassay without chromatography. However, upon separation of the steroids by chromatography it was found that much of the androgen present was in fact dihydrotestosterone not testosterone. Both TU and dihydrotestosterone undecanoate were detected in plasma by gas chromatography and it is suggested that the ester is absorbed as such from the intestine and the unesterified steroid subsequently released by hydrolysis. The convenience of oral administration, the resulting prolonged elevated plasma androgen levels and the probable lack of deleterious effects on the liver may render oral TU of value where androgen replacement therapy is indicated.

We report a case of a calcified liver tumor in a 23 year old female patient who presented with virilization and a mild degree of Cushing's syndrome. Androgen levels were elevated; there was loss of cortisol circadian rhythm and marked increase in urinary <em>17</em>-ketogenic and <em>17</em>-<em>ketosteroids</em> which failed to suppress with administration of dexamethasone. Venous sampling by inferior vena cava catheterization showed that the highest steroid hormone levels were in blood from the right hepatic vein. After death, in vitro studies revealed that the tumor contained testosterone and cortisol as determined by immunofluorescence techniques. The adrenals and ovaries were atrophic. Results of metyrapone testing indicated dyshormonogenesis. To our knowledge, this is the first case of an adrenal rest tumor of the liver proved to be functionally active.

Multidirectional changes in the natural history of many cardiovascular syndromes have been linked to different levels of daily and monthly geomagnetic activity (GMA). Previous studies have found that in periods of high GMA, there were more admissions for acute myocardial infarction and more cases of anterior wall myocardial infarction. Results also indicated: higher out-patient mortality and a trend towards higher hospital mortality from acute myocardial infarction; higher diastolic arterial pressure in healthy subjects and in treated hypertensive patients; higher prolactin and <em>17</em>-corticosteroid levels in the peripheral blood; more severe migraine attacks and more admissions for CVA and cerebrovascular insufficiency in male patients; changes in many blood coagulation cellular gradients (platelet count, basophils in the peripheral blood), a rise in platelet aggregation, fibrinogen level and a drop in leukocyte adhesiveness. Periods of low GMA showed a related increase (negative correlation) in in-hospital non-myocardial infarction-related cardiovascular deaths. Only in times of lowest GMA did inferior wall myocardial infarction exceed anterior wall myocardial infarction. Low GMA was also associated with higher levels of growth hormone and 11-<em>ketosteroids</em> in the peripheral blood, more sudden deaths, some increase in electrical heart instability number of ventricular and supraventricular extrasystoles and higher rate of ventricular tachycardia. The monthly occurrence of pregnancy-induced hypertension was negatively correlated with GMA level. Gender differences were noted in some of the parameters. Other studied parameters did not show changes related to GMA. These included hemoglobin level, electrolyte level, heart beat and pulse rate. Moreover, some observed cardiovascular fluctuations that were related to the level of GMA, also showed differences in the rising and dropping parts of the 11-year cycle of solar activity. It has been suggested that some of the changes observed in many clinical syndromes may be related to the concomitant activation of the serotoninergic system.

We have cloned an approx. 5-kb fragment of Pseudomonas testosteroni DNA containing the structural gene of delta 5-3-<em>ketosteroid</em> isomerase into the EcoRI site of the lambda gt11 genome. Escherichia coli infected with these recombinant phages produce a polypeptide which is recognized by antiserum raised against the purified isomerase. Four of the recombinant lambda gt11 clones contain significant levels of isomerase activity and produce an immunopositive polypeptide of the same apparent Mr as the native isomerase obtained from P. testosteroni. The approx. 5-kb fragment hybridizes to synthetic 21-mer and <em>17</em>-mer oligodeoxynucleotide mixtures corresponding to the 5' and 3' regions, respectively, of the expected nucleotide sequence of the gene.

A 49-year-old woman with virilization demonstrated biochemical features traditionally ascribed to virilizing ovarian tumors: marked elevation of serum testosterone level with normal urinary excretion of <em>17</em>-<em>ketosteroids</em> and normal serum dehydroepiandrosterone level. An adrenal cortical adenoma containing neoplastic cells indistinguishable from Leydig cells, including the demonstration of characteristic crystalloids of Reinke, was shown to be the source of the elevated testosterone level. A review of the literature revealed 13 cases with a similar biochemical profile, and of these, two were reported to contain crystalloids of Reinke. Taking cognizance of the fact that these characteristic crystalloids are only found in 40 percent of Leydig cell tumors of the testis, it is concluded that Leydig cells may be present in, and may be active participants in, the pathophysiology of a number of testosterone-secreting adrenal tumors.

A technique of monolayer tissue culture of human fetal adrenal cells was developed in order to study steroidogenic responses to factors such as ACTH. The daily production of 12 steroids [pregnenolone, <em>17</em>-hydroxy pregnenolone, dehydroepiandrosterone (DHA), DHA sulfate, progesterone, <em>17</em>-hydroxyprogesterone, androstenedione, testosterone, corticosterone, 11-desoxycortisol, cortisol, and aldosterone) was measured by RIA. Initially, fresh fetal adrenal cells produced DHA, DHA sulfate, <em>17</em>-hydroxypregnenolone, and small amounts of cortisol, but in the absence of ACTH, the production of all steroids declined during culture to low levels. The addition of physiological amounts (1-10(4) pg/ml) of either alpha ACTH-1(1-24) or alpha ACTH-(1-39) or coculture with fetal pituitary cells elicited a progressive rise in steroid production during the first 4-6 days of incubation. The lowest ACTH doses elicited a proportionately greater adrenal androgen response (as reflected in the DHA to cortisol ratio), but with increasing ACTH dosage, there was greater stimulation of cortisol production, which equalled or exceeded that of DHA. The data demonstrate that fetal adrenal cells may be maintained in short term culture and can respond to physiological amounts of ACTH. The progressive increase in the production of cortisol and other delta 4, 3-<em>ketosteroids</em> in vitro suggests that the characteristic fetal pattern of steroidogenesis may result from the interaction of ACTH with some circulating inhibitor of adrenal 3 beta-hydroxysteroid dehydrogenase.