Anabolic Steroid Abuse Research Articles

Investigations into the feasibility of routine ultra high performance liquid chromatography–tandem mass spectrometry analysis of equine hair samples for detecting the misuse of anabolic steroids, anabolic steroid esters and related compounds

The detection of the abuse of anabolic steroids in equine sport is complicated by the endogenous nature of some of the abused steroids, such as testosterone and nandrolone. These steroids are commonly administered as intramuscular injections of esterified forms of the steroid, which prolongs their effects and improves bioavailability over oral dosing. The successful detection of an intact anabolic steroid ester therefore provides unequivocal proof of an illegal administration, as esterified forms are not found endogenously. Detection of intact anabolic steroid esters is possible in plasma samples but not, to date, in the traditional doping control matrix of urine. The analysis of equine mane hair for the detection of anabolic steroid esters has the potential to greatly extend the time period over which detection of abuse can be monitored.Equine mane hair samples were incubated in 0.1M phosphate buffer (pH 9.5) before anabolic steroids (testosterone, nandrolone, boldenone, trenbolone and stanozolol), anabolic steroid esters (esters of testosterone, nandrolone, boldenone and trenbolone) and associated compounds (fluticasone propionate and esters of hydroxyprogesterone) were extracted by liquid–liquid extraction with a mix of hexane and ethyl acetate (7:3, v:v). Further sample clean up by solid phase extraction was followed by derivatisation with methoxylamine HCL and analysis by UHPLC–MS/MS. Initial method development was performed on a representative suite of four testosterone esters (propionate, phenylpropionate, isocaproate and decanoate) and the method was later extended to include a further 18 compounds.The applicability of the method was demonstrated by the analysis of mane hair samples collected following the intramuscular administration of 500mg of Durateston® (mixed testosterone esters) to a Thoroughbred mare (560kg). The method was subsequently used to successfully detect boldenone undecylenate and stanozolol in hair samples collected following suspicious screening findings from post-race urine samples. The use of segmental analysis to potentially provide additional information on the timing of administration was also investigated.

Effect of androgenic-anabolic steroids and heavy strength training on patellar tendon morphological and mechanical properties

Combined androgenic-anabolic steroids (AAS) and overloading affects tendon collagen metabolism and ultrastructure and is often associated with a higher risk of injury. The aim of this prospective study was to investigate whether such effects would be reflected in the patellar tendon properties of individuals with a history of long-term resistance training and AAS abuse (RTS group), compared with trained (RT) and untrained (CTRL) nonsteroids users. Tendon cross-sectional area (CSA), stiffness, Young's modulus, and toe limit strain were measured in vivo, from synchronized ultrasonography and dynamometry data. The patellar tendon of RT and RTS subjects was much stiffer and larger than in the CTRL group. However, stiffness and modulus were higher in the RTS group (26%, P < 0.05 and 30%, P < 0.01, respectively) than in the RT group. Conversely, tendon CSA was 15% (P < 0.05) larger in the RT group than in RTS, although differences disappeared when this variable was normalized to quadriceps maximal isometric torque. Yet maximal tendon stress was higher in RTS than in RT (15%, P < 0.05), without any statistical difference in maximal strain and toe limit strain between groups. The present lack of difference in toe limit strain does not substantiate the hypothesis of changes in collagen crimp pattern associated with AAS abuse. However, these findings indicate that tendon adaptations from years of heavy resistance training are different in AAS users, suggesting differences in collagen remodeling. Some of these adaptations (e.g., higher stress) could be linked to a higher risk of tendon injury.

Caution for Anabolic Androgenic Steroid Use: A Case Report of Multiple Organ Dysfunction Syndrome

We report a 42-year-old male amateur body builder and user of anabolic androgenic steroids, who developed ARDS, acute kidney injury, and refractory supraventricular tachycardia. He required extracorporeal membrane oxygenation, continuous veno-venous hemodialysis, and catheter ablation. We believe that long-term anabolic androgenic steroid abuse predisposed the patient to multiple organ dysfunction syndrome, from its immunomodulatory effects in an otherwise healthy patient. Anabolic androgenic steroid use should be part of the history taking process, since it may complicate diagnosis, disease progression, and prognosis.

Modulation of Follistatin and Myostatin Propeptide by Anabolic Steroids and Gender

The purpose of this pilot study was to investigate the impact of training, anabolic steroids and endogenous hormones on myostatin-interacting proteins in order to identify manipulations of myostatin signalling. To identify whether analysis of the myostatin interacting proteins follistatin and myostatin propeptide is suitable to detect the abuse of anabolic steroids, their serum concentrations were monitored in untrained males, bodybuilders using anabolic steroids and natural bodybuilders. In addition, we analysed follistatin and myostatin propeptide serum proteins in females during menstrual cycle. Our results showed increased follistatin concentrations in response to anabolic steroids. Furthermore, variations of sex steroid levels during the menstrual cycle had no impact on the expression of follistatin and myostatin propetide. In addition, we identified gender differences in the basal expression of the investigated proteins. In general, follistatin and myostatin propeptide concentrations were relatively stable within the same individual both in males and females. In conclusion, the current findings provide an insight into gender differences in myostatin-interacting proteins and their regulation in response to anabolic steroids and endogenous hormones. Therefore our data provide new aspects for the development of doping prevention strategies.

A supraphysiological dose of testosterone induces nitric oxide production and oxidative stress

Accumulating evidence indicates that abuse of anabolic androgenic steroids may cause cardiovascular adverse side-effects, including endothelial dysfunction. The aim of the present study was to investigate the effects of supra-physiological doses of testosterone on the endothelial production of nitric oxide (NO) and oxidative stress in vitro and in vivo. Testosterone enanthate was administrated as of a single 500 mg dose to healthy volunteers (N = 27). Gene expression was studied in human vascular endothelial cells exposed to testosterone. The in vivo results show that the urinary NO level and the antioxidative capacity were significantly decreased two days after testosterone administration. In agreement, our in vitro studies show that testosterone inhibits the gene expression of endothelial NO synthase (eNOS) after 48 hours. When the antioxidant seleno-L-methionine was added, the down-regulation of mRNA specific eNOS was partly abrogated. The mRNA expression of antioxidizing enzyme genes was significantly inhibited after eight hours and recovered 48 hours after testosterone treatment of endothelial cells. These results show that a supraphysiological dose of testosterone decreases the expression of eNOS and consequently the formation of NO, which could partly be explained by oxidative stress. These results indicate that supraphysiological doses of testosterone may induce endothelial dysfunction, which is of interest in relation to the cardiovascular adverse side-effects observed in anabolic androgenic steroid abusers.

Endocrine fingerprint analyses as an alternative strategy to detect the abuse of anabolic steroids

In doping analysis anabolic substances are usually detected by GCMS and LCMS. However, even substances with a chemical structure typical for this class of substances are sometimes not identified when their exact chemical structure is unknown. Therefore, indirect detection techniques may help to complement the classic detection methods in the future. For this purpose the effects of anabolic steroid abuse on the serum concentrations of different hormones were analyzed. Blood and urine samples from a group of male bodybuilders (n = 10) and a male control group (n = 6, 29.7 ± 9.35 years, 81.92 ± 4.18 kg bw, 183 ± 5 cm height) were collected for this study. The urine samples were subsequently analyzed for metabolites of anabolic steroids by the Institute of Biochemistry, German Sport University Cologne. The positively tested bodybuilders were allocated in the group abusing bodybuilders (ABB, n = 5, 24.2 ± 6.3 years, 102 ± 13.77 kg bw, 184 ± 7 cm height) and the negatively tested in the group natural bodybuilders (NB, n = 5, 34.4 ± 14.57 years, 94.5 ± 10.47 kg bw, 182 ± 7 cm height). The serum concentration of cortisol, thyroid-stimulating hormone (TSH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in comparison to inhibin, thyroxin (T4), and insulin-like growth factor 1 (IGF-1) was measured by ELISA or RIA. In the ABB T4 and prolactin serum conc. were significantly reduced and the LH conc. significantly enhanced compared to the control group. The TSH serum conc. was significantly reduced in the ABB and NB group compared to the control group. For inhibin a significantly reduced serum conc. in ABB compared to the control group and a tendency toward a difference compared to NB could be observed. The results of this pilot study demonstrate that the comparative analysis of serum concentrations may be suitable to detect doping abuse: TSH as a marker for physical activity; LH, prolactin and inhibin as potential markers for steroid abuse.

Reference ranges for the urinary steroid profile in a Latin‐American population

The urinary steroid profile has been used in clinical endocrinology for the early detection of enzyme deficiencies. In the field of doping, its evaluation in urine samples is used to diagnose the abuse of substances prohibited in sport. This profile is influenced by sex, age, exercise, diet, and ethnicity, among others; laboratories own reference ranges might compensate for ethnic differences among population and inter-laboratory biases. This paper shows the reference ranges obtained in the Antidoping Laboratory of Havana for the following steroid profile parameters: ten androgens (testosterone, epitestosterone, androsterone, etiocholanolone, 5α-androstan-3α,17β-diol, 5β-androstan-3α,17β-diol, dehydroepiandrosterone, epiandrosterone, 11β-hydroxyandrosterone and 11β-hydroxyetiocholanolone), three estrogens (estradiol, estriol and estrone), two pregnanes (pregnanediol and pregnanetriol) and two corticosteroids (cortisol and tetrahydrocortisol). The urine samples (male: n = 2454 and female: n = 1181) and data obtained are representative of population from Latin-American countries like Cuba, Venezuela, Mexico, Dominican Republic, Guatemala and Chile. Urine samples were prepared by solid-phase extraction followed by enzymatic hydrolysis and liquid-liquid extraction with an organic solvent in basic conditions. Trimethylsilyl derivatives were analyzed by gas chromatography coupled to mass spectrometry. Reference ranges were established for each sex, allowing the determination of abnormal profiles as a first diagnostic tool for the detection of the abuse of androgenic anabolic steroids. The comparison with the Caucasian population confirms that the urinary steroid profile is influenced by ethnicity.

Brain Nerve Growth Factor Unbalance Induced by Anabolic Androgenic Steroids in Rats

Anabolic androgenic steroids (AAS) are synthetic androgen-like compounds that are abused in sport communities despite their adverse effects. Nerve growth factor (NGF) influences neuronal differentiation and survival, and it also mediates higher brain functions such as learning and memory. Changes in NGF expression have been implicated in neurodegenerative disorders, including Alzheimer disease. Hence, we decided to study the effect of chronic AAS exposure on brain NGF profile, NGF-dependent cholinergic function, and related behavioral performance. Male Wistar rats were injected for 4 wk with either nandrolone or stanozolol at daily doses (5.0 mg·kg(-1), s.c.) that are considered equivalent to those abused by humans. NGF levels and NGF receptor (TrkA and p75NTR) expression were measured in the hippocampus and in the basal forebrain. Choline acetyltransferase expression was evaluated in basal forebrain. Spatial learning and memory were assessed using the Morris water maze. AAS treatment caused region-specific changes in the expression of NGF and its receptors. Both nandrolone and stanozolol increased NGF levels in the hippocampus and reduced NGF levels in the basal forebrain, reduced p75NTR expression in the hippocampus, and failed to affect TrkA expression in the basal forebrain. Finally, AAS treatment reduced the expression of choline acetyltransferase in the basal forebrain and impaired the behavioral performance in the Morris water maze. The evidence that supraphysiological doses of AAS cause neurotrophic unbalance and related behavioral disturbances raises the concern that AAS abuse in humans may affect mechanisms that lie at the core of neuronal plasticity.

The application of health belief model among body building athletes in the identification of belief associated with anabolic steroid use

Background: Progressive prevalence of anabolic steroid use and physical and mental consequences of these drugs are known as worldwide healthy problems. Willingness to being fitness and having muscular body and unhealthy beliefs about anabolic steroids are determinant behavioral factors of anabolic storied use. The purpose of this study was to determine the behavioral reason of anabolic steroid abuse as well as to assess the relationship between anabolic steroid use and perceived threat of anabolic steroid use among body building athletes. Methods: This study is a cross-sectional one in which a total of 300 body builders, were randomly selected for participation in Hamadan. A structured questionnaire was applied for collecting data about anabolic steroids abuse, self efficacy not to use, perceived susceptibility and seriousness of anabolic steroids use. The data were analyzed using SPSS-13. Results : seventy three athletes (24.3%) reported that they have used anabolic steroid. Anadrol was the mostly used substance among participants. Most important reason mentioned by athletes for consuming anabolic steroids was body fitness. Perceived seriousness (OR= 0/675 & p-value = 0/000) and perceived self efficacy (OR= 0/796 & p-value = 0/000) were two important predicting factors for using anabolic steroid. Conclusion: Planning and implementation of educational program for the increasing of self efficiency and informing athletes of the serious side effects of consuming these drugs would have a positive effect on preventing or reducing their consumption.

Adolescent ischemic stroke associated with anabolic steroid and cannabis abuse

We report on a 16-year-old body builder who suffered from an acute ischemic stroke. In the urine, cannabis metabolites as well as metabolites of the oral androgenic-anabolic steroid methandrostenolone were detected, both known to be associated with stroke events. This report highlights the role of cannabis and steroid abuse that induce strokes in the absence of arteriopathy, cardioembolism or thrombophilia. Owing to new upcoming socio-behavioral aspects of late childhood and early adolescent life, this formally rare abuse of cannabis and/or anabolic steroids as well as their associations with strokes becomes more current than ever.

Neuroprotective effect of erythropoietin on nandrolone decanoate-induced brain injury in rats

Anabolic–androgenic steroids (AAS) are used in the medical treatment of many disorders. Erythropoietin (EPO) is a hematopoietic cytokine that has anti-apoptotic, anti-oxidative, and anti-inflammatory effects. The aim of the present study is to investigate the neuroprotective effects of EPO in the hippocampus, parietal cortex and prefrontal cortex, in brain damage due to nandrolone decanoate. 35 Wistar male rats were randomly divided into: (1) control group, (2) sham group, (3) nandrolone decanoate group (ND, intramuscular, 10mg/(kgweek), 8 weeks), (4) ND+low dose EPO treated group (ND+L-EPO) and (5) ND+high dose EPO treated group (ND+H-EPO). EPO was administrated by intraperitoneal injection at a dose of 100U/(kgday) for L-EPO treatment and at a dose of 500U/(kgday) for H-EPO treatment during 8 weeks. The number of neurons of CA1, CA2, CA3 and dentate gyrus of hippocampus, parietal cortex and prefrontal cortex were significantly less in the ND group compared with the control group. Treatment with H-EPO significantly preserved the number of neurons in hippocampus when compared with ND administrated. Besides, H-EPO treatment decreased the number of TUNEL-positive and active caspase-3 positive cells and MDA levels and increased GPx levels when compared to ND group. In conclusion, abuse of AAS causes reduction in the number of neurons in hippocampus, parietal cortex and prefrontal cortex regions and increases oxidative damage and therefore H-EPO may be useful as a neuroprotective agent in brain injury.

Misconceptions About Anabolic-Androgenic Steroid Abuse

CME Educational Objectives 1. Recognize which patient populations are most likely to abuse anabolic-androgenic steroids (AAS). 2. Identify and understand the frequently underestimated adverse symptoms of AAS. 3. Become familiar with ways patients avoid detection of AAS use. Although anabolic-androgenic steroids were widely used by elite athletes as early as the 1950s, it was not until the 1980s that anabolic-androgenic steroid abuse began to spread beyond the elite athletic world and into the general population. Because widespread anabolic-androgenic steroid abuse is so new, many clinicians and researchers are not nearly as familiar with this form of substance abuse as they are with other, more venerable drugs.

Echocardiographic findings in power athletes abusing anabolic androgenic steroids.

Anabolic androgenic steroids (AAS) abuse for improving physical appearance and performance in body builders is common and has been considered responsible for serious cardiovascular effects. Due to disagreement about cardiovascular side effects of these drugs in published articles, this case control study was designed to evaluate the echocardiographic findings in body builder athletes who are current and chronic abusers of these drugs. Body builder athletes with continuous practice for the preceding two years and were training at least twice weekly were selected and divided into AAS abuser and non user and compared with age and BMI matched non athletic healthy volunteers (15 cases in each group). There was no significant difference in left ventricular size or function either systolic or diastolic in comparison to cases and control groups. The only difference was in diastolic size of septum and free wall but observed differences were only significant (P = 0.05) between first (athletic with AAS abuser) and third group (non athletic and nonuser). The difference between the above-mentioned indexes were not significant between two groups of athletes. Observed differences in diastolic size of septum and free wall is in favor of that long term abuse of anabolic steroid results in accentuation of physiologic hypertrophy due to long term sport most probably due to higher rate pressure product. Furthermore long term abuse and supra pharmacologic doses do not have significant effect in size and left ventricular function.

Serotonin modulates anxiety-like behaviors during withdrawal from adolescent anabolic–androgenic steroid exposure in Syrian hamsters

From the U.S. to Europe and Australia anabolic steroid abuse remains high in the adolescent population. This is concerning given that anabolic steroid use is associated with a higher incidence of pathological anxiety that often appears during withdrawal from use. This study uses pubertal Syrian hamsters (Mesocricetus auratus) to investigate the hypothesis that adolescent anabolic/androgenic steroid (AAS) exposure predisposes hamsters to heightened levels of anxiety during AAS withdrawal that is modulated by serotonin (5HT) neural signaling. In the first two sets of experiments, adolescent AAS-treated hamsters were tested for anxiety 21days after the cessation of AAS administration (i.e., during AAS withdrawal) using the elevated plus maze (EPM), dark/light (DL), and seed finding (SF) tests and then examined for differences in 5HT afferent innervation to select areas of the brain important for anxiety. In the EPM and DL tests, adolescent AAS exposure leads to significant increases in anxiety-like response during AAS withdrawal. AAS-treated hamsters showed long-term reductions in 5HT innervation within several areas of the hamster brain implicated in anxiety, most notably the anterior hypothalamus and the central and medial amygdala. However, no differences in 5HT were found in other anxiety areas, e.g., frontal cortex and lateral septum. In the last experiment, adolescent AAS-treated hamsters were scored for anxiety on the 21st day of AAS withdrawal following the systemic administration of saline or one of three doses of fluoxetine, a selective serotonin reuptake inhibitor. Saline-treated hamsters showed high levels of AAS withdrawal-induced anxiety, while treatment with fluoxetine reduced AAS withdrawal-induced anxiety. These findings indicate that early AAS exposure has potent anxiogenic effects during AAS withdrawal that are modulated, in part, by 5HT signaling.

Prevalence of behaviour-related fertility disorders in a clinical sample: results of a pilot study

There is no doubt that lifestyle factors can be detrimental to fertility. The aim of the present pilot study was to identify initial prevalence rates for behaviour-related fertility disorders in a clinical sample of couples wanting a child. Between February 2010 and August 2010, all patients coming for the first time to Heidelberg University's Women's Hospital for consultation on involuntary childlessness were asked to fill out a questionnaire designed by the authors of this article. The questionnaire was based on a review of the relevant literature, with special reference to the latest research findings on behaviour detrimental to fertility. Of the 156 couples addressed, 110 women and 100 men took part in the study. For behaviour-related infertility, 9 % of the women and 3 % of the men in our sample were classified on the basis of BMI <18.5, sexual disorders, or abuse of anabolic steroids. If we include smokers, these figures increase: 11 % female smokers and 18 % male smokers. A further 19 % of the women practised sport to an excessive degree; and 26 % of the women and 53 % of the men had a BMI ≥25. The prevalence of behaviour-related fertility disorders should not be underestimated. For the prevention of behaviour-related fertility disorders, it is important to inform the population about lifestyle-mediated fertility risks.

Cardiovascular risk and androgenic anabolic steroids

Although several drugs are purported to improve exercise performance, androgenic anabolic steroids (AAS) are the most widespread. Furthermore, unlike other drugs, their use has expanded beyond competition, to non-competitive and recreational athletes. Correspondingly health professionals are more likely to come into contact with users of AAS than with users of other performance-enhancing drugs. While there are numerous reports outlining serious cardiovascular consequences to high-dose AAS abuse, this evidence is often limited by difficulties in gaining access to users due to the legal status of AAS. Additionally the co-abuse of other substances (as additional muscle mass enhancers, or to mitigate possible side effects) is a further confounding factor. This review examines the evidence for AAS having a negative effect on the cardiac and vascular tissue and the corresponding risk of developing cardiovascular disease. Possible mechanisms of action by which AAS bring about these changes are also discussed.

Cardiac effects of anabolic steroid use amongst recreational body builders - a CMR study

Background Abuse of anabolic steroids to induce skeletal muscle hypertrophy is widespread amongst recreational bodybuilders, however, the cardiac effects of such drugs have not been systematically documented. The ability of cardiovascular magnetic resonance (CMR) to image the myocardium in any plane and achieve full myocardial coverage renders it the gold standard for assessing LV volumes and mass.We sought to investigate the cardiac effects of anabolic steroid use with CMR hypothesising that significant hypertrophy and distinct LV remodelling would be seen in steroid users relative to non-users. Methods

Research in Androgenic Anabolic Steroid Abuse

Research in Androgenic Anabolic Steroid Abuse

Development of Hepatocellular Carcinoma Associated with Anabolic Androgenic Steroid Abuse in a Young Bodybuilder: A Case Report

Introduction. Many different etiological factors are involved in the development of hepatocellular carcinoma (HCC). We report the case of HCC in a 37-year-old male professional bodybuilder with extensive anabolic androgenic (AAS) steroid abuse. Case Presentation. Because of increasing epigastric and abdominal pain, abdominal ultrasound was performed in a 37-year-old male professional bodybuilder. A hyperechoic lesion in the liver was detected in segment VI. The magnetic resonance imaging showed hepatomegaly and confirmed the lesion, which showed features of a hepatocellular adenoma (HCA). Laboratory values were inconspicuous. After laparoscopic segmentectomy the histological examination revealed HCC. Conclusion. While the development of HCA in the liver by chronic intake of AAS is well known, little is known about the association with HCC. The presented case may indicate aetiological association of chronic intake of AAS and the development of HCC.

Short QT interval is unreliable marker of anabolic androgenic steroid abuse in competitive athletes

Previous animal and human studies provided the evidence that testosterone may affect ventricular repolarization by shortening of the QT interval. Synthetic derivatives of testosterone, modified to enhance its anabolic properties, are occasionally abused by some competitive athletes. We assessed whether the QT interval duration could discriminate androgenic anabolic steroids (AAS)-using strength athletes (SA) from drug-free endurance athletes (EA), by comparing 25 formulas for QT interval correction. We recruited 22 elite male athletes involved in long-term strength or endurance training and 20 sedentary controls. All elite SA reported AAS abuse, whereas EA and controls were AAS-free. AAS-using SA had markedly shorter QT-interval than AAS-free EA (348 +/- 42.3 vs. 400 +/- 34.2 ms; p < 0.001). Also, drug-free EA had a significantly longer QT-interval than sedentary persons (400 +/- 34.2 vs. 358 +/- 18.9 ms; p < 0.01). In contrast, no significant difference in the QT-interval duration was observed between AAS users and control group (348 +/- 42.3 vs. 358 +/- 18.9 ms; p = 0.394). After the QT interval was adjusted for heart rate (HR) according to 25 different formulas, only the Ashman equation yielded considerable differences among the groups that were in line with those observed before correction. Inconsistent results obtained by different correction formulas along with inability to discriminate QT (QTc) interval duration between AAS-misusing athletes and control group do not support the use of QT (QTc) interval for anti-doping purposes.