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Anabolic-androgenic steroids: effects on social behavior and baseline heart rate.

Examined the effects of anabolic-androgenic steroids (AS) on behavior, baseline heart rate (HR), and stress-induced HR responses. Twenty-four cynomolgus monkeys were assigned to four mixed social groups of both AS and sham control animals. For 2 months, AS-treated monkeys received biweekly injections of testosterone, and, on an identical schedule, the control animals were injected with a sham solution. Behavioral data revealed that AS disrupted the social milieu such that all dominant animals exhibited increases in dominant behavior and subordinates manifested increased submission. These changes returned to pretest levels 8 weeks following termination of the drug intervention. Affiliative behaviors decreased on the part of all animals as a function of AS and, with the exception of play behavior, failed to return to pretest levels after the 8-week period of recovery. AS created an increase in baseline HR, particularly among the dominant animals. Interestingly, the subordinate AS animals experienced a decrease in baseline HR response. AS had no apparent influence on stress-induced HR reactivity.

Agonistic Behavior↗

Synthetic androgens suppress the transformed phenotype in the human prostate carcinoma cell line LNCaP.

Experiments have been designed to investigate hormonal effects on the human prostatic carcinoma cell line LNCaP in the presence of complete foetal calf serum. At physiological concentrations (3.3 x 10(-9)M), several derivatives of 17 alpha-methyl-testosterone led to a significant reduction of cell proliferation, inhibition of colony formation in soft agar, change of morphology, induction of a prostate specific mRNA and down-regulation of c-myc RNA. Two different antiandrogens, hydroxyflutamide and cyproterone acetate, were capable of reversing the effects exerted by the synthetic androgens on growth properties. The proliferation rate of control cells devoid of androgen receptor was not inhibited by synthetic androgens. Our results indicate that the cellular androgen response mechanism of LNCaP cells is intact and that synthetic androgens elicit androgen receptor mediated suppression of the transformed phenotype. Rare cases of remission of prostatic cancer on androgen treatment have been reported. LNCaP cells may be a model of an uncommon class of prostatic cancer which responds favourably to androgen treatment.

Antigens, Neoplasm↗

Correlation of clinical and in vitro erythropoietic responses to androgens in renal failure.

To assess in vitro erythropoietic cultures as a tool to discriminate between patients whose anemia improves with and those whose anemia fails to improve with androgens, bone marrows of 24 anemic patients receiving maintenance hemodialysis in hormone-depleted plasma clots were cultured. While erythroid colony proliferation in the presence and absence of androgens by cells of seven patients with good clinical outcomes who were initially studied was similar to that of normal donors, it was reduced (P less than 0.001) in cultures of cells from seven patients whose clinical responses were ultimately poor. Fewer than 25 colonies/6 X 10(4) cells at optimal erythropoietin concentrations, and less than a 25% enhancement in colony growth by androgens were observed in only those cultures derived from clinical nonresponder marrows. Using these criteria, nine of ten prospectively classified patients (five clinical responders and four nonresponders) were identified correctly.

Adult↗

Differentiation enhances aminolevulinic acid-dependent photodynamic treatment of LNCaP prostate cancer cells.

Photodynamic therapy using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) may be applied to the treatment of neoplasms in a variety of organs. In order to enhance existing regimens of photodynamic therapy, we investigated the effects of adding differentiation therapy to photodynamic therapy in human prostate cancer cells in vitro. The objective of differentiation therapy per se is to reverse the lack of differentiation in cancer cells using pharmacological agents. The motivation for this study was to exploit the differentiation-dependent expression of some heme enzymes to enhance tumour cell toxicity of ALA-photodynamic therapy. A short course of differentiation therapy was applied to increase PpIX formation during subsequent ALA exposure. Using the synthetic androgen R1881, isomers of retinoic acid, and analogues of vitamin D for 3 to 4 days, exogenous ALA-dependent PpIX formation in LNCaP cells was increased, along with markers for growth arrest and for differentiation. As a consequence of higher PpIX levels, cytotoxic effects of visible light exposure were also enhanced. Short-term differentiation therapy increased not only the overall PpIX production but also reduced that fraction of cells that contained low PpIX levels as demonstrated by flow cytometry and fluorescence microscopy. This study suggests that it will be feasible to develop protocols combining short-term differentiation therapy with photodynamic therapy for enhanced photosensitisation.

Aminolevulinic Acid↗

Regulation of expression of Na+,K+-ATPase in androgen-dependent and androgen-independent prostate cancer.

The beta 1-subunit of Na+,K+-ATPase was isolated and identified as an androgen down-regulated gene. Expression was observed at high levels in androgen-independent as compared to androgen-dependent (responsive) human prostate cancer cell lines and xenografts when grown in the presence of androgens. Down-regulation of the beta 1-subunit was initiated at concentrations between 0.01 nM and 0.03 nM of the synthetic androgen R1881 after relatively long incubation times (> 24 h). Using polyclonal antibodies, the concentration of beta 1-subunit protein, but not of the alpha 1-subunit protein, was markedly reduced in androgen-dependent human prostate cancer cells (LNCaP-FGC) cultured in the presence of androgens. In line with these observations it was found that the protein expression of total Na+,K+-ATPase in the membrane (measured by 3H-ouabain binding) was also markedly decreased. The main function of Na+,K+-ATPase is to maintain sodium and potassium homeostasis in animal cells. The resulting electrochemical gradient is facilitative for transport of several compounds over the cell membrane (for example cisplatin, a chemotherapeutic agent experimentally used in the treatment of hormone-refractory prostate cancer). Here we observed that a ouabain-induced decrease of Na+,K+-ATPase activity in LNCaP-FGC cells results in reduced sensitivity of these cells to cisplatin-treatment. Surprisingly, androgen-induced decrease of Na+,K+-ATPase expression, did not result in significant protection against the chemotherapeutic agent.

Androgens↗

Repression of androgen receptor mediated transcription by the ErbB-3 binding protein, Ebp1.

Members of the ErbB family of receptors have been implicated in regulation of androgen receptor (AR) activity. Ebp1, an ErbB-3 binding protein recently cloned in our laboratory, possesses an LXXLL motif important in mediating interactions with nuclear hormone receptors. Therefore, we sought to determine if Ebp1 could bind AR and influence AR transcriptional activation potential. We demonstrate in this study that Ebp1 bound to AR in vitro and in vivo, and that this binding was increased by androgen treatment. The C terminal 79 amino acids of Ebp1 were sufficient to bind AR. The N terminal domain of AR was responsible for binding Ebp1. Ligand-mediated transcriptional activation of both artificial and natural AR regulated promoters was inhibited by ectopic expression of ebp1 in transient transfection systems. Ebp1 deletion mutants that either lacked the C terminal AR binding region or had a mutated LXXLL motif failed to inhibit AR activated transcription. PSA expression from its endogenous promoter was also decreased in LNCaP prostate cancer cells overexpressing Ebp1. The growth of AR positive LNCaP cells was inhibited by ectopic expression of ebp1, but mutants that failed to repress transcription did not inhibit cell growth. These studies suggest that Ebp1 may play a role in the function of the AR and provide a link between ErbB receptors and the AR.

Adaptor Proteins, Signal Transducing↗

Interleukin-4 enhances prostate-specific antigen expression by activation of the androgen receptor and Akt pathway.

Androgen receptor (AR) plays an important role in the development and progression of prostate cancer upon the action of androgen through the binding of the androgen-responsive elements (AREs) on the target genes. Abnormal activation of the AR by nonandrogen has been implicated in the progression of androgen-independent prostate cancer. The levels of interleukin-4 (IL-4) are significantly elevated in sera of patients with hormone refractory prostate cancer. The potential role of IL-4 on the activation of AR was investigated in prostate cancer cells. IL-4 enhances AR-mediated prostate-specific antigen (PSA) expression and ARE-containing gene activity through activation of the AR in the androgen ablation condition in human prostate cancer cells. The AR can also be sensitized by IL-4 and activated by significantly lower levels of androgen (10 pM of R1881) in prostate cancer cells. IL-4 enhances nuclear translocation of AR and increases binding of the AR to the ARE in LNCaP prostate cancer cells. Blocking of the Akt pathway by an Akt-specific inhibitor LY294002 abrogates IL-4-induced PSA expression and AR signaling. These results demonstrate that IL-4 enhances PSA expression through activation of the AR and Akt signaling pathways in LNCaP prostate cancer cells. Understanding IL-4-induced signaling leading to abnormal activation of AR will provide insights into the molecular mechanisms of androgen-independent progression of prostate cancer cells.

Androgen Antagonists↗

Cytosolic androgen receptor in regenerating rat levator ani muscle.

The development of the cytosolic androgen receptor was studied after degeneration and regeneration of the rat levator ani muscle after a crush lesion. Muscle regeneration appears to recapitulate myogenesis in many respects. It therefore provides a model tissue in sufficiently in large quantity for investigating the ontogenesis of the androgen receptor. The receptor in the cytosol of the normal levator ani muscle has binding characteristics similar to those of the cytosolic receptor in other androgen-sensitive tissues. By day 3 after a crush lesion of the levator ani muscle, androgen binding decreased to 25% of control values. This decrease was followed by a 4-5 fold increase in hormone binding, which attained control values by day 7 after crush. Androgen binding remained stable at the control value up to day 60 after crushing. These results were correlated with the morphological development of the regenerating muscle after crushing. It is concluded that there is little, if any, androgen receptor present in the early myoblastic stages of regeneration; rather, synthesis of the receptor may occur after the fusion of myoblasts and during the differentiation of myotubes into cross-striated muscle fibres.

Anal Canal↗

Effect of methyl testosterone administration on plasma viscosity in postmenopausal women.

BACKGROUND: Coronary heart disease (CHD) is the leading cause of mortality in women, with an incidence that increases after menopause, hence suggesting a cardioprotective role of oestrogen. Menopause also results in a decline in androgen levels with resulting symptoms of decreased libido and sexual dysfunction. Recently, there has been a growing interest in the treatment of postmenopausal women with androgens. However, no data are available on plasma viscosity and fibrinogen levels in postmenopausal women on combined oestrogen/androgen therapy. METHODS: We conducted a randomized, double-blind, parallel-group 16-week study evaluating the effects of methyltestosterone supplementation on plasma viscosity and fibrinogen levels in postmenopausal women already on oestrogen replacement therapy (ERT) for at least 3 months. Women 21 years and older who were menopausal (natural or surgical) for at least 12 months were enrolled in the study. Participants were randomized to (1) an oestrogen-only group taking 1.25 mg esterified oestrogen (E-group) and (2) an oestrogen plus methyltestosterone (1.25 mg esterified oestrogen and 2.5 mg methyltestosterone) group (EA-group). Progesterone was not administered during the study period and women with intact uteri were given medroxyprogesterone 10 mg daily for 14 days at the completion of the study. RESULTS: After 16 weeks of treatment, both groups had a significant increase in serum oestradiol levels from baseline. The levels of total oestrogen were significantly higher in the E-group compared to the EA-group (P < 0.001). There was a greater decrease in the LH and SHBG levels in the EA-group (P = 0.01). There was no difference in total testosterone; however, free testosterone levels were significantly higher in the EA-group (P = 0.01). At the end of the study, there was a significant decrease in plasma viscosity only in the EA-group (P = 0.01). Fibrinogen levels increased in both the groups, reaching significance only in the EA-group (P = 0.006). Baseline weight, body mass index (BMI) and the duration of menopausal status did not have any significant impact on the changes in plasma viscosity or fibrinogen. Women in the EA-group showed significant reductions in total cholesterol (P = 0.009), high density lipoprotein (HDL) (P < 0.001) and triglyceride (TG) levels (P = 0.001). There was no significant change in these parameters in the E-group. CONCLUSION: This prospective study shows that the treatment of postmenopausal women on oestrogen with low-dose oral methyltestosterone results in a significant reduction in plasma viscosity. This lowering of plasma viscosity was achieved despite an increase in fibrinogen levels. Significant lowering of lipoproteins, especially TG levels, might have been responsible for this benefit. The combination regimen did not result in major side-effects. Based on these results, we feel confident in recommending low-dose androgens to postmenopausal women with a history of sexual dysfunction and decreased libido.

Aged↗

Androgen administration in middle-aged and ageing men: effects of oral testosterone undecanoate on dihydrotestosterone, oestradiol and prostate volume.

The gradual reduction of plasma testosterone in middle-aged and older men from mid-life onwards coincides paradoxically with the time when there is progressive growth of the prostate, a highly androgen-dependent organ. The growing interest in androgen therapy for older men makes it essential to understand the effects of exogenous testosterone on the non-diseased prostate, yet few studies are available. The present study examined prostate volume, prostate-specific antigen (PSA) and lower urinary tract symptom (IPSS) score in 207 men, aged 40-83 years, presenting with clinical features of age-related androgen deficiency [sexual and/or urinary dysfunction, elevated lutenizing hormone (LH)] who were treated for 6 months with oral testosterone undecanoate (TU). Men were divided into two groups, group 1 (n=92, plasma testosterone levels > 13 nmol/L) were treated with 80 mg daily; group 2 (n=115, plasma testosterone levels < 13 nmol/L) were treated with given 120 mg daily. Before treatment and after 1, 3 and 6 months of treatment, prostate volume was measured by ultrasound and hormones [testosterone, dihydrotestosterone, oestradiol, LH, follicle-stimulating hormone (FSH)] and PSA were measured. Within 1 month of treatment, the elevated blood LH levels were markedly decreased in all men in group 1, as well as most men in group 2. Group 2 was subdivided into men whose LH levels were suppressed (n=95, group 2a) and those whose LH levels did not suppress (n=20, group 2b). Men in group 1 and 2a had marked decreases in prostate volume, PSA and lower urinary tract symptom (IPSS) scores whereas no significant changes were observed in group 2b. Groups 1 and 2a also had more striking suppression of LH, FSH, dihydrotestosterone and oestradiol whereas group 2b had no significant increases in blood testosterone concentrations. These findings suggest that exogenous testosterone in middle-aged and older men with some clinical features of age-related androgen deficiency can retard or reverse prostate growth and that elevated plasma LH may be a useful index of severity of age-related androgen deficiency.

Adult↗

Androgen-sensitive human prostate cancer cells, LNCaP, produce both N-terminally mature and truncated prostate-specific antigen isoforms.

To characterize prostate-specific antigen (PSA) produced by cancer cells, different isoforms of PSA secreted by the human prostate cancer cells, LNCaP, were purified. LNCaP-PSA production was induced by synthetic androgen, R1881. LNCaP-PSA was separated into four pools. The molecular mass of LNCaP-PSA isoforms in these pools was 34 kDa under reducing conditions and 29 kDa under nonreducing conditions on SDS/PAGE. pI of LNCaP-PSA isoforms varied from 6.8 to 8.2. Pool A had the highest specific activity, 37 nmol/(min x mg). All the pools formed stable complexes with alpha1-antichymotrypsin and alpha2-macroglobulin. The pools contained 10-60% of N-terminally correctly processed LNCaP-PSA isoforms. According to the molecular modelling, the addition or deletion of two or four N-terminal amino acids could affect the three-dimensional structure and thereby remarkably reduce the enzyme activity of LNCaP-PSA.

Amino Acid Sequence↗

Muscle strength and ageing: methodological aspects of isokinetic dynamometry and androgen administration.

1. To evaluate interventions aiming to increase muscle strength in older men, it is necessary to use an objective and reproducible method to measure strength. The most reliable method to evaluate muscle strength is isokinetic peak torque (PT) measured by a dynamometer; however, raw PT varies with differences in body size and an optimal scaling for body physique to control confounding effects of body size is necessary to make the most valid comparisons. 2. The present study was designed to estimate the effects of age (part A) and androgen administration (part B) on muscle strength and to estimate reproducibility and evaluate various scaling methods in order to optimize comparisons of isokinetic PT measurements. 3. A single isokinetic exercise protocol was used to compare the muscle strength of 31 healthy men of two age groups (< 40 and > 60 years; part A) and change in strength due to administration of dihydrotestosterone (DHT; 70 mg/day) or placebo gel for 3 months in 35 healthy older (> or = 60 years) men (part B). 4. Muscle strength was assessed by a total of 16 PT measurements using a Cybex NORM dynamometer (Cybex, Ronkonkoma, NY, USA). Age-related differences in muscle strength were estimated by using PT evaluated as raw data or scaled by normalizing methods, including simple ratio (PT/weight), allometric PT (PT/weight(0.67)) and adjustment of PT by weight, height, body mass index and body surface area (BSA). The goodness-of-fit for various scaling methods was compared using the Akaike Information Criterion (AIC) as an objective measure of model-based entropy reduction. 5. The effects of DHT administration according to different scaling methods were estimated by eta-squared measure of effect size in treatment models. In part A, older men were weaker than younger men in five knee PT, consistently by all eight analysis models but not in shoulder PT. In part B, DHT treatment resulted in an increase one knee PT (dominant knee flexion at 120 degrees/s) with the difference consistent in all seven models. 6. The scaling model using BSA proved superior to other comparison models throughout both parts of the present study according to entropy minimization criteria (AIC) for goodness-of-fit of the model or eta-squares for treatment effect size. 7. We conclude that differences in muscle strength due to age or androgen administration in older men are restricted to a minority of lower limb contractions and that use of BSA scaling for PT values is considered the best scaling method for muscle strength comparisons in either cross-sectional or longitudinal studies.

Adult↗

Effects of dehydroepiandrosterone on rat apolipoprotein AI gene expression in the human hepatoma cell line, HepG2.

Serum apolipoprotein AI (apoAI) levels correlate with the risk of developing atherosclerosis. Previous studies have suggested that dehydroepiandrosterone (DHEA) lowers high-density lipoprotein (HDL)-cholesterol levels. We investigated whether or not DHEA may lower HDL-cholesterol levels by suppressing apoAI gene transcription in hepatocytes. ApoAI mRNA levels, assessed by Northern blotting, were suppressed in HepG2 cells treated with DHEA (34%) (10 microg/mL) or testosterone (36%) (T, 1 microg/mL). Estradiol alone (E2, 1 microg/mL) had relatively little effect on apoAI mRNA levels, while E2 in combination with DHEA prevented a decrease in apoAI mRNA levels compared to DHEA alone. To determine whether these effects were due to changes in apoAI gene transcription, HepG2 cells were transfected with a plasmid carrying the full-length promoter of the rat apoAI gene ligated into a chloramphenicol acetyltransferase (CAT) reporter construct. The plasmid pCMV.SPORT-beta-gal was included in each transfection to normalize the data to transfection efficiency. Cells were then cultured in the presence or absence of DHEA (10 microg/mL), T (1 microg/mL), 17alpha-methyltestosterone (MTT, 1 microg/mL), 5alpha-dihydrotestosterone (DHT, 1 microg/mL), E2 (1 microg/mL), or a combination of DHEA plus E2, T plus E2, MTT plus E2, and DHT plus E2, for 24 hours. CAT activity, relative to beta-galactosidase activity, was reduced by 19.6%, 57.6%, 38.6%, and 54.6% with DHEA, T, DHT, and MTT addition, respectively. E2 increased CAT activity by 43.8%. When the androgens (ie, DHEA, T, DHT, or MTT) were combined with E2, apoAI promoter activity was suppressed. We conclude, therefore, that androgens downregulate apoAI promoter activity in the presence or absence of E2. However, the changes in mRNA levels do not always reflect changes in promoter activity, suggesting that these steroids may have additional post-transcriptional effects on steady-state apoAI mRNA levels. It remains to be established if the transcriptional effects we observed are mediated through an androgen response element.

Animals↗

Assignment of anabolic-androgenic and antiandrogenic properties to some chlorine-substituted steroids on the basis of their binding characteristics to the androgen receptor of the rat seminal vesicle.

In this study we investigated the affinity of several 4-chlorinated and 1-ene derivatives of 17 alpha-methyltestosterone (MT) and 17 alpha-methyl-5 alpha-dihydrotestosterone (MDHT) to the androgen receptor, and, additionally, the effect of a few MT-derived steroids on the activity of the 5 alpha-reductase enzyme present in the rat seminal vesicle. From our results we conclude, that delta 1 or/and delta 4 double bonds in ring A counteract the inhibition of receptor-binding caused by chlorine-substitution at C4; the dissociation of myotropic and androgenic effects [= M/A dissociation] of 4-chloro-MT (as compared to MT) is due to its inactivation by 5 alpha-reductase in androgen target organs and/or to the inhibition of the conversion of endogenous testosterone to DHT; the M/A dissociation of 1-ene-MT and 4-chloro-1-ene-MT may be explained by their inability to be activated by 5 alpha-reductase; for the same reason, M/A dissociation can be assigned to the effects of 4 alpha-chloro-1-ene-DHT. We determined the short-term and long-term competition of cyproterone acetate and chlormadinone acetate with [3H]DHT for receptor binding at 0 degrees C and showed, that the complexes formed by these antiandrogens with the androgen receptor have equally reduced stabilities compared to the DHT-receptor complex.

Anabolic Agents↗

Ontogeny of an 8S androgen binding protein in rat liver cytosol.

Studies were performed to elucidate the ontogeny of a single class of androgen binding protein in male rat liver cytosol which exhibits characteristics of a ligand specific, high affinity (Kd = 2.3 nM), 8S-receptor capable of nuclear translocation. Detectable levels of receptor first appear at 45 days of age in the male and reach maximum concentration at 65 days. Barely detectable levels are seen in females throughout the duration of study (80 days). Gonadectomy in both sexes (65 days) and androgen treatment of oophorectomized females do not alter the normal development of sexual differentiation of the high affinity androgen receptor. After neonatal castration (2 days) and DES replacement however, receptor sites do not undergo differentiation and adult males exhibit female levels. Conversely, neonatal androgen replacement in 2-day castrates partially restores the level of binding sites to control males values (TP, 71%; DHT, 51%). Neonatal castration without replacement retards but does not fully eliminate sexual differentiation of levels of receptor sites in adult males. Likewise, neonatal androgen treatment in females results in a partial masculinization of binding sites. Following hypophysectomy, levels of receptor sites in females are similar to intact or hypophysectomized males; sexual differences in the adult are abolished. These studies suggest that sexual differentiation of specific liver cytosol androgen binding sites in the adult may be partially programmed at birth by testicular androgen and furthermore, adult sexual dimorphism is maintained through an inhibitory influence of the pituitary in the female.

Androgen-Binding Protein↗

[Pathological rupture of the distal biceps tendon after long-term androgen substitution].

QUESTION: Is the long-standing use of androgenes able to cause tendon lesions with pathological tendon ruptures? METHOD: In a case of a rupture of the distal biceps tendon after long-standing testosterone-substitution it is tried to show the connection between the use of androgenes and pathological tendon rupture by the help of the patients treatment documents, the X-rays and sonograms, the histological findings and the results of the clinical examination. RESULTS: For the first time a case with a rupture of the distal biceps tendon after long-standing testosterone-substitution for the support of a genital transformation is described. Since other tendon damaging factors could be excluded the suspicion of tendon alteration caused by androgenes is obvious. CONCLUSION: Looking at tendon ruptures of professionals and even amateur sportsmen the possible connection between long-standing use of androgenes and tendon damage has to be considered.

Adult↗

Hepatic tumors induced by sex steroids.

Our study of more than 250 women with hepatic tumors, accessioned in our tumor registry at the University of Louisville, disclosed three types of tumor: FNH , HCA, and HCC. The ingestion of sundry kinds of sex steroids by the majority of these women, chiefly for purposes of preventing conception, warrants the suspicion that such hormones induced these different types of hepatic tumors. Publications by others reporting similar hepatic tumors in men using male sex steroids lends support to this hypothesis. Rupture of the hepatic tumor and consequent hemorrhage, producing hemoperitoneum, is a major risk factor. Other presenting symptoms are pain and palpable mass. Symptomatic women using OCs should be subjected to a CT or technetium hepatic scan as an initial screening assessment. Because of the imminent possibility of rupture, large turgid vascular tumors should be resected without biopsy. Biopsy-proved HCC should also be removed surgically. All other tumors, including small multiple tumors, will usually regress when exogenous sex steroids are withdrawn and pregnancy avoided. Other significant hepatic changes observed in this study are peliosis hepatis, periportal sinusoidal dilation, and vascular lesions. The branches of the hepatic artery and the tributaries of the portal vein show combinations of intimal and smooth muscle proliferation, vascular thickening, occlusive intimal thickening, and, at times, obstructing thrombosis. Similar smooth muscle proliferation in the afferent vessels of the livers of animals treated with sex steroids suggests that there is a cause and effect relationship in women using OCs.

Adolescent↗