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PubMed2003

The effect of anabolic steroids on mandibular growth.

The aim of this study was to assess the effect of nandrolone (Deca-Durabolin, AKZO Nobel, Cambridge, United Kingdom) on mandibular growth in juvenile and adult rats with radiographic cephalometry and immunoradiology. Juvenile (n = 16) and adult (n = 16) inbred female Wistar-Kyoto rats were compared. Each group was divided into 2 subgroups with 8 experimental (E) and 8 control (C) animals in each subgroup. Lateral headfilms taken before and after the 70-day study period were analyzed. Body weight and blood serum IGF-I levels were monitored weekly. The results showed marked mandibular growth changes in both the juvenile and the adult E rats. Body weight increase was larger in the E than in the C animals. The IGF-I blood serum levels were similar in the juvenile E and C rats but higher in the adult E animals than in the adult C animals. It was found that the anabolic steroid (Deca-Durabolin) had a significant effect on mandibular growth in both juvenile and adult rats.

Age Factors↗
PubMed1990

Gas chromatographic-mass spectrometric confirmation of 19-nortestosterone in the urine of untreated boars--effect of the administration of Laurabolin.

The presence of 17 beta-19-nortestosterone (nandrolone, NT, 17 beta-19-NT) and its epimer 17 alpha-19-nortestosterone (epiNT, 17 alpha-19-NT) was investigated in the urine of six untreated boars, obtained from experimental farms. The presence of 17 beta-19-nortestosterone was screened by RIA and HPTLC and confirmed by GC-MS analysis. Additionally, the two epimers (NT and epiNT) were investigated in the urine of a boar (two-year-old miniature male pig weighing 50 kg) before and after injection of 100 mg Laurabolin (nortestosterone laurate, Intervet N.V., Belgium). The isolation of the steroids was based on sample clean-up with solid phase extraction and subsequent high-performance liquid chromatography. Both gas chromatographic retention data and mass spectrometric data (selected ion monitoring and full spectrum) were used for detection and identification. The presence of 17 beta-19-nortestosterone in the urine of the boars that were not injected proves the endogenous production of the steroid. The absence of the 17 alpha-epimer in the urine of the injected boar suggests that 17 alpha-19-nortestosterone is not a major metabolite of 17 beta-19-nortestosterone.

Animals↗
PubMed2005

Contamination of dietary supplements and positive drug tests in sport.

The use of dietary supplements is widespread in sport and most athletes competing at the highest level of competition use some form of dietary supplementation. Many of these supplements confer no performance or health benefit, and some may actually be detrimental to both performance and health when taken in high doses for prolonged periods. Some supplements contain excessive doses of potentially toxic ingredients, while others do not contain significant amounts of the ingredients listed on the label. There is also now evidence that some of the apparently legitimate dietary supplements on sale contain ingredients that are not declared on the label but that are prohibited by the doping regulations of the International Olympic Committee and of the World Anti-Doping Agency. Contaminants that have been identified include a variety of anabolic androgenic steroids (including testosterone and nandrolone as well as the pro-hormones of these compounds), ephedrine and caffeine. This contamination may in most cases be the result of poor manufacturing practice, but there is some evidence of deliberate adulteration of products. The principle of strict liability that applies in sport means that innocent ingestion of prohibited substances is not an acceptable excuse, and athletes testing positive are liable to penalties. Although it is undoubtedly the case that some athletes are guilty of deliberate cheating, some positive tests are likely to be the result of inadvertent ingestion of prohibited substances present in otherwise innocuous dietary supplements.

Androgens↗
PubMed2004

Analytical strategies for the detection of non-labelled anabolic androgenic steroids in nutritional supplements.

Recent studies showed that non-hormonal supplements such as vitamins, minerals and amino acids can contain anabolic androgenic steroids not declared on the labels of the products. These undeclared substances (often prohormones of testosterone or 19-nortestosterone) can cause health risks to consumers and might lead to positive results in sports doping control, especially for the nandrolone metabolite norandrosterone. The analysis of nutritional supplements for anabolic steroids has proven to be rather difficult due to the different matrices in the various products. To conduct a broad-based analysis, a few robust methods capable of analysing various matrices are needed. To obtain a sensitive gas chromatography-mass spectrometry (GC-MS) analysis, a method including extraction and purification of the analytes followed by GC-MS analysis of the trimethylsilyl (TMS) derivatives of the steroids was developed. The limit of detection was improved by the addition of a mixture of 1-N,N-diisopropylamino-n-alkanes (DIPAs) to the final extract. In pure creatine monohydrate powder the limits of detection were demonstrated to be 0.1 ng microg (-1) for dehydroepiandrosterone (DHEA) and estr-4-ene-3beta,17beta-diol, 0.7 ng g(-1) for 5alpha-androstane-3beta,17beta-diol and androsta-1,4-diene-3,17-dione, 1 ng g(-1) for estr-5-ene-3beta,17beta-diol, estr-4-ene-3,17-dione, 19-nortestosterone, androst-4-ene-3, 17-dione and testosterone, and 2 ng g(-1) for androst-4-ene-3beta,17beta-diol and androst-5-ene-3beta,17beta-diol. The recovery (determined at 200 ng g(-1)) ranged from 32% for 19-nortestosterone to 92% for androst-5-ene-3beta,17beta-diol. During the investigation of different nutritional supplements, several analytical difficulties occurred. Aspects of homogenization, extraction, separation, derivatization and GC-MS measurement as well as strategies for the solution of problems arising were optimized. For quantitative measurements of the steroids in nutritional supplements, deuterated internal standards of the specific steroids or standard addition are necessary to compensate for matrix effects.

Alkanes↗
PubMed2005

Validation of a GC-MS screening method for anabolizing agents in aqueous nutritional supplements.

A sensitive and selective method for the screening of anabolizing agents in aqueous nutritional supplements is described and validated. A total of 28 different anabolizing agents are screened for, including testosterone and prohormones, nandrolone and prohormones, stanozolol, and metandienone. The different analytes are extracted from the aqueous nutritional supplements by liquid-liquid extraction with a mixture of pentane and freshly distilled diethylether (1:1) after the supplements have been made alkaline with a NaHCO3-K2CO3 (2:1) buffer. The anabolizing agents are derivatized with a mixture of MSTFA-NH4I-ethanethiol (320:1:2) as routinely used for the screening of anabolic steroids extracted from urine. The derivatives are analyzed by gas chromatography (GC)-mass spectrometry (MS) in the selective ion monitoring mode. The limits of detection range from 1 to 10 ng/mL. One aqueous nutritional supplement (creatine serum) was analyzed with this screening method and was found to contain dehydroepiandrosterone (DHEA) at very low concentrations. The presence of DHEA could be confirmed with GC-MS-MS. Results of the application of this method and a similar method for solid nutritional supplements previously described are given.

Anabolic Agents↗
PubMed2000

Morphological effects of an anabolic steroid on muscle fibres of the diaphragm in mice.

Until recently, rats have been used for studies on the effects of anabolic steroids (AS) for improving physical performance. However, no consistent results on the changes in muscle fibre volume have ever been obtained. In this study we investigated the morphological effects of nandrolone phenylpropionate, an AS, on the muscle fibres of the diaphragm in adult male mice. AS (5 mg kg(-1) week(-1)) was injected intramuscularly in the experimental group (AS-administration group), and peanut oil (0.08 ml week(-1)) was injected in the control group. The cross-sectional areas of Type I (red) and Type II (white and intermediate) muscle fibres after 4 weeks of AS administration were 135% and 139% larger than those in the control group, respectively. The mean cross-sectional areas of mitochondria in the subsarcolemmal (SS) region and interfibrillar (IF) region in Type I fibres and the SS region of the Type II muscle fibres in the AS-administration group were 139%, 135% and 124% larger than those in the control group, respectively. No significant differences in the cross-sectional area of mitochondria were noted between the group of mice administered with AS for 8 weeks and the control group, showing that the effect of the AS administration attained a peak at 4 weeks, but that the effect of the 8 weeks of AS administration declined. We concluded from these results that the response of Type I muscle fibres to AS is stronger than that of Type II muscle fibres, as is the response of SS mitochondria compared to IF mitochondria. We also suggested that the two types of mitochondria may have different roles and that administration of AS may cause a different response in the two types of mitochondria.

Anabolic Agents↗
PubMed1980

Corticoids and human skin fibroblasts: intracellular specific binding in relation to growth inhibition.

The binding of 3H-triamcinolone acetonide to soluble macromolecules of cultured human skin fibroblasts was studied in an attempt to explain the mechanism underlying the inhibitory effects of glucocorticoids on cell growth. The results were as follows: Cultured human skin fibroblasts contain in cytosol a high affinity binding system for glucocorticoids. Various glucocorticoid derivatives competed for specific binding of 3H-triamcinolone acetonide. In some but not all instances this competition was related to the clinical efficacy of the derivatives under study and to their potency for the inhibition of cell growth. A specific glucocorticoid binding system was detectable in steroid-sensitive, low-density cell cultures (apparent Bmax = 200 fmoles/mg protein). The number of steroid binding sites was lower in high-density cell cultures (apparent Bmax = 125 fmoles/mg protein). The sensitivity to growth inhibition by glucocorticoids was markedly decreased in the high-density cell cultures. There were no differences in the affinity constants between these cell cultures (Kdiss. = 3.3 X 10-9 M). When cells were grown in medium containing glucocorticoid, renewal of the incubation medium led to disappearance of the growth-inhibitory effects, whereas specific binding was not affected. Nandrolone, an inhibitor of cell growth, abolished the growth-inhibitory effects of glucocorticoids but did not displace 3H-triamcinolone acetonide from its binding sites. The results suggest that in addition to a mechanism mediated by a glucocorticoid binding system with receptor like properties also other factors as well appear of relevance for the control of cell growth. These factors may be beyond the actual binding process of steroid and involve the action at the level of genomic expression of the cell.

Animals↗
PubMed1977

The effect of various treatments on the size of sebaceous glands of hairless mice and hairless hamsters.

The effect of castration on the size of sebaceous glands of male hairless mice and male hairless hamsters was studied over a period of 4 weeks. Statistically significant decreases in the size of sebaceous glands were observed. The effects of intracutaneous injections of micronized crystalline suspensions of testosterone, testosterone propionate, testosterone phenylpropionate, testosterone decanoate and nandrolone phenylpropionate on the sebaceous glands of castrated hairless mice, castrated hairless hamsters and intact female hairless hamsters were studied over a period of one week. Highly significant increases in sebaceous gland volume were observed 6-7 days after treatment. The effects of subcutaneous and intracutaneous injections of micronized crystalline suspensions of testosterone on the sebaceous glands of intact male and female hairless hamsters were examined at sites local and distal to the injection. Increases in sebaceous gland volume were limited to local sites in the male but increases were observed locally and distally in the female. There was no difference between subcutaneous and intracutaneous injections.

Animals↗
PubMed1983

Diffusion barriers in skin--a new method of comparison.

Using a method which allows the net barrier potential of skin to be expressed in terms of a 'penetration half-life', skin barriers to nandrolone and allyl oestrenol penetration were compared in human, rat and hamster skin. The ranking order in terms of the net barrier was: human = haired rat greater than or equal to hairless rat greater than hairless hamster. Using the same principles, the net barrier effects of the various layers in whole human skin were also compared. The dermis was only slightly less effective than the stratum corneum as a barrier to diffusion.

Allylestrenol↗
PubMed1977

Evidence of decarboxylation of lysine by mammalian ornithine decarboxylase.

In enzymic preparations from mouse kidney stimulated with the anabolic steroid Durabolin (nandrolone phenpropionate) lysine and ornithine were shown to inhibit the decarboxylation of each other competitively. The Michaelis constants for the decarboxylations were approximately equal to the inhibition constants of the two amino acids. The pH optima of the decarboxylation of lysine and ornithine were found to be identical. Chromatographic studies of the enzyme preparation on a Sephadex G-150 Superfine column did not bring about a separation of the two enzyme activities. The ratio of the decarboxylating activities was practically the same during the elution. Lysine decarboxylating activity was also shown to be present in growth hormone stimulated rat liver. The results are in agreement with the assumption that the decarboxylation of lysine and ornithine is carried out by the same enzyme.

Animals↗
PubMed1984

Changes in neuromuscular performance and muscle fiber characteristics of elite power athletes self-administering androgenic and anabolic steroids.

The influence of androgenic-anabolic steroid-induced changes in measures of body composition, muscle fiber characteristics and various aspects of the neuromuscular performance of the leg extensor muscles was investigated in five experimental and six control power athletes during the 24-week programmed strength training followed by the additional six week training without hormone drugs. The mean values of the dosages of self-administration during the 24-week period were 31.0 +/- 14.3 mg/day for anabolic steroids (methandienone, stanozolol, nandrolone) and 178.4 +/- 82.7 mg/week for testosterone. During the 24-week hormone period the experimental group gained in fat-free weight (p less than 0.01) and in the mean muscle fiber areas (p less than 0.01) of the vastus lateralis muscle while the corresponding gains in the control group were minor (NS). The increases of maximal isometric force in the experimental and control groups were 14.7% (p less than 0.01) and 6.1% (NS), respectively, and the values obtained in average load-vertical jumping height curves were improved significantly (p less than 0.05) only in the experimental group. Increases of 18.2% (p less than 0.001) and 12.9% (p less than 0.01) took place in the squat lift in the experimental and control groups, respectively. Both groups demonstrated similar (p less than 0.05) improvements in isometric fast force production. During the additional six week programmed training without hormone drugs significant (p less than 0.05) increases were observed in the experimental group in addition to maximal isometric force and the squat-lift but also in isometric fast force production, while the corresponding changes in the control group were minor (NS). It is suggested that strength training in combination with administration of androgenic-anabolic steroids causes improvements in selected neuromuscular parameters. These changes may be greater than those of caused by the strength training alone.

Adult↗
PubMed2003

Glucuronidation of anabolic androgenic steroids by recombinant human UDP-glucuronosyltransferases.

A multidimensional study on the glucuronidation of anabolic androgenic steroids and their phase I metabolites by 11 recombinant human UDP-glucuronosyltransferases (UGTs) was carried out using liquid chromatographic-tandem mass spectrometric analyses. Large differences between the enzymes with respect to the conjugation profiles of the 11 tested aglycones were detected. Two UGTs, 1A6 and 1A7, did not exhibit measurable activity toward any of the aglycones that were examined in this study. Regioselectivity was demonstrated by UGTs 1A8, 1A9, and 2B15 that preferentially catalyzed hydroxyl glucuronidation at the 17beta-position. Most of the other enzymes glucuronidated hydroxyl groups at both the 3alpha- and the 17beta-positions. Clear stereoselectivity was observed in glucuronidation of diastereomeric nandrolone metabolites (5alpha-estran-3alpha-ol-17-one and 5beta-estran-3alpha-ol-17-one), whereas such specificity was not seen when analogous methyltestosterone metabolites were assayed. UGTs 1A1, 1A3, 1A4, 1A8, 1A9, 1A10, 2B4, 2B7, and 2B15 readily glucuronidated 5alpha-androstane-3alpha,17beta-diol, but none of them exhibited methyltestosterone glucuronidation activity. In agreement with the latter observations, we found that the methyltestosterone glucuronidation activity of human liver microsomes is extremely low, whereas in induced rat liver microsomes it was significantly higher. The homology among UGTs 1A7 to 1A10 at the level of amino acid sequence is very high, and it was thus surprising to find large differences in their activity toward this set of aglycones. Furthermore, the high activity of UGT1A8 and 1A10 toward some of the substrates indicates that extrahepatic enzymes might play a role in the metabolism of anabolic androgenic steroids.

Anabolic Agents↗
PubMed2002

Steroid receptor concentration in aged rat hindlimb muscle: effect of anabolic steroid administration.

Skeletal muscle is a target of anabolic steroid action; however, anabolic steroid's affect on aged skeletal muscle is not well understood. The effect of 4 wk of nandrolone decanoate (ND) administration on hindlimb muscles of 5- and 25-mo-old Fischer 344/Brown Norway rats was examined. ND (6 mg/kg body wt) was injected every 7th day for 4 wk. Controls received an oil injection. ND significantly reduced 25-mo-old rat perirenal fat pad mass by 30%. Soleus (Sol) and plantaris (Plan) muscle-to-body weight ratios were reduced in 25-mo-old rats. ND did not affect Sol or Plan muscle-to-body weight ratios at either age. Sol DNA concentration was reduced by 25% in 25-mo-old rats, and ND increased it to 12% greater than 5-mo-old rats. ND did not affect Plan DNA content. Sol androgen receptor (AR) protein in 25-mo-old rats was reduced to 35% of 5-mo-old values. ND increased AR protein by 900% in 25-mo-old rat Sol. Plan AR concentration was not affected by aging but was induced by ND in both age groups. Aging or ND treatment did not affect glucocorticoid receptor levels in either muscle. These data demonstrate that fast- and slow-twitch rat hindlimb muscles differ in their response to aging and ND therapy.

Aging↗
PubMed2000

Anabolic steroids induce region- and subunit-specific rapid modulation of GABA(A) receptor-mediated currents in the rat forebrain.

Anabolic-androgenic steroids (AAS) have become significant drugs of abuse in recent years with the highest increase reported in adolescent girls. In spite of the increased use of AAS, the CNS effects of these steroids are poorly understood. We report that in prepubertal female rats, three commonly abused AAS, 17alpha-methyltestosterone, stanozolol, and nandrolone, induced rapid and reversible modulation of GABAergic currents in neurons of two brain regions known to be critical for the expression of reproductive behaviors: the ventromedial nucleus of the hypothalamus (VMN) and the medial preoptic area (mPOA). All three AAS significantly enhanced peak synaptic current amplitudes and prolonged synaptic current decays in neurons of the VMN. Conversely all three AAS significantly diminished peak current amplitudes of synaptic currents from neurons of the mPOA. The endogenous neuroactive steroids, 3alpha-hydroxy-5alpha-pregnan-20-one and 5alpha-androstane-3alpha,17beta-diol, potentiated currents in the VMN as did the AAS. In contrast to the negative modulation induced by AAS in the mPOA, the endogenous steroids potentiated responses in this region. To determine the concentration response relationships, modulation by the AAS, 17alpha-methyltestosterone (17alpha-meT), was assessed for currents evoked by ultrafast perfusion of brief pulses of GABA to acutely isolated neurons. Half-maximal effects on currents elicited by 1 mM GABA were elicited by submicromolar concentrations of AAS for neurons from both brain regions. In addition, the efficacy of 10(-5) to 10(-2) M GABA was significantly increased by 1 microM 17alpha-meT. Previous studies have demonstrated a striking dichotomy in receptor composition between the VMN and the mPOA with regard to gamma subunit expression. To determine if the preferential expression of gamma(2) subunit-containing receptors in the VMN and of gamma(1) subunit-containing receptors in the mPOA could account for the region-specific effects of AAS in the two regions, responses elicited by ultrafast perfusion of GABA to human embryonic kidney 293 cells transfected with alpha(2), beta(3), and gamma(2) or alpha(2), beta(3), and gamma(1) subunit cDNAs were analyzed. As with native VMN neurons, positive modulation of GABA responses was elicited for alpha(2)beta(3)gamma(2) recombinant receptors, while negative modulation was induced at alpha(2)beta(3)gamma(1) receptors as in the mPOA. Our data demonstrate that AAS in doses believed to occur in steroid abusers can induce significant modulation of GABAergic transmission in brain regions essential for neuroendocrine function. In addition, the effects of these steroids can vary significantly between brain regions in a manner that appears to depend on the subunit composition of GABA(A) receptors expressed.

Anabolic Agents↗
PubMed1976

Studies on membrane-bound renin in the mouse and rat.

Membrane-bound renin was isolated from the submaxillary gland and kidney of the white Swiss Webster mouse and from the kidney of the rat. Administration of the male sex hormone testosterone or an anabolic steroid, such as nandrolone phenpropionate (Durabolin), induces renin activity in microsomal fraction of the submaxillary gland of the young female mouse. Actinomycin D blocks this induction partially. Renin that sedimented with the microsomal fraction was isolated from the homogenized kidney of the mouse by density gradient centrifugation. Microsomal renin was also isolated from rat kidney disrupted by the N2 cavitation technique. The enzyme in the microsomal fraction that had been washed extensively was only partially active, but freezing and thawing or Triton X-100 activated microsomal renin. The same microsomal fraction contained active kallikrein. In pilot studies a plasma membrane-enriched fraction was isolated from the microsomal fraction of the homogenized rat kidney cortex that had both renin and kallikrein activity.

Animals↗
PubMedDate not supplied

Histologic and histochemical study of effects of anabolic steroids on the female larynx.

The larynges of 30 unmated, mature female albino mice were examined histologically and histochemically after injecting them with 19-nandrolone phenylpropionate. Permanent irreversible changes, in the form of parakeratosis and squamous metaplasia of the epithelium, and hypertrophy of the muscle fibers, were observed together with persistence of increased alkaline phosphatase enzyme intensity in the endothelium of capillaries and increased activity of nonspecific esterase in the muscles. Similar changes may be responsible for the alteration in the human female's voice after prolonged treatment with anabolic steroids.

Anabolic Agents↗
PubMed1992

Bone and mineral metabolism in aged male rats: short and long term effects of androgen deficiency.

Both short and long term effects of androgen deficiency and steroid replacement therapy on skeletal homeostasis were investigated in aged (13-month-old) male rats. The animals were either sham operated (n = 28) or orchidectomized (orch; n = 89). The orch animals were divided into 5 groups; 26 rats received an empty sc Silastic implant (orch), all others received an implant containing testosterone (T), 5 alpha-dihydrotestosterone (DHT), 17 beta-estradiol (E2), or nandrolone (Nandro; 15-16 rats in each group). Half of the rats were killed 1 month (short term experiment) after implantation; the others were killed 4 months after implantation (long term experiment). Short term androgen deficiency caused a significant increase in both serum osteocalcin and histomorphometric parameters of bone turnover measured at the proximal tibial metaphysis, but not in a significant decrease in bone mass at this site. This increase in bone turnover was prevented not only by T and DHT, but also by E2 and Nandro. Long term androgen deficiency resulted in a decrease in the calcium content of both tibia and lumbar vertebrae. Cancellous bone volume in the proximal tibial metaphysis was +/- 50% lower in the orch group (P less than 0.001) 4 months after orchidectomy. At the same time, cortical bone was lost in orch rats; femoral cortical thickness was reduced by 12% (P less than 0.01), and cortical density tended to be lower. T, DHT, E2, or Nandro treatment completely prevented this decrease in cortical thickness and density. T and Nandro were also able to prevent the cancellous bone loss. DHT could only partly prevent cancellous bone loss. E2 treatment resulted not only in a sustained decrease in both serum osteocalcin concentrations and histomorphometric indices of bone turnover, but also in a net gain of cancellous bone volume (P less than 0.05 vs. sham). No significant differences in serum concentrations of vitamin D metabolites or nephrogenous cAMP were observed between groups in both short and long term experiments. We conclude that bone mass in aged male rats was significantly decreased 4 months after orchidectomy, preceded by an early increase in bone turnover. Both the early increase in bone turnover and the later decrease in bone mass were prevented by aromatizable and nonaromatizable androgens by estrogen and by nandralone.

Aging↗
PubMed1985

A specific radioimmunoassay for the detection of 19-nortestosterone in urine.

The antisera used were raised in rabbits against a 19-nortestosterone-7 alpha-carboxyethylthioether conjugate of bovine serum albumin. Tritium-labeled nortestosterone was used as tracer. Cross reactivities with metabolites of nandrolone, other anabolics and endogenous steroids were very low. To achieve additional specificity a clean up and separation procedure was developed, using isocratic high performance liquid chromatography. So far no qualitative discrepancies have been observed between the assay described here and confirmative determinations by combined high performance liquid chromatography-gas chromatography-mass spectrometry.

Animals↗