Anabolic steroids and extreme violence: a case of murder after chronic intake and under acute influence of metandienone and trenbolone.

A 32-year-old male went to the police to claim he just killed his girlfriend by inflicting several stabs with a kitchen knife. He was very nervous and particularly aggressive. About 90 min after the assault, a blood specimen was collected with natrium fluoride as preservative. The blood was free of alcohol, pharmaceuticals and drugs of abuse, but tested positive by LC-MS/MS for metandienone (32 ng/mL) and trenbolone (9 ng/mL). The perpetrator admitted regular consumption of anabolic steroids to enhance his muscular mass, as he was a professional security agent. To document long-term steroid abuse, a hair specimen was collected 3 weeks after the assault, which tested positive for both drugs. Segmental analyses revealed in the proximal 1.5 cm segment, corresponding to the period of the assault, the simultaneous presence of metandienone (11 pg/mg) and trenbolone (14 pg/mg), while only metandienone (3 pg/mg) was identified in the distal 1.5 cm segment. As aggressiveness and violence can be associated with abuse of anabolic steroids, the aetiology of this domestic crime was listed to be due impulsive behaviour in a context of antisocial lifestyle.

A jaundiced bodybuilder Cholestatic hepatitis as side effect of injectable anabolic-androgenic steroids.

The use of anabolic steroids is prevalent in recreational athletes. This case report describes a young amateur bodybuilder who was referred to our outpatient clinic with jaundice and loss of appetite due to cholestatic hepatitis. Additional tests including a liver biopsy made it likely that the hepatitis was caused by the injectable anabolic steroid trenbolone enanthate. Cholestatic hepatitis may not be limited to the use of oral anabolic-androgenic steroids, as is widely assumed. Therefore, and because of other side effects, the recreational use of all forms of anabolic steroids should be discouraged.

Studies of the pharmacokinetic profile, in vivo efficacy and safety of injectable altrenogest for the suppression of oestrus in mares.

OBJECTIVE: To investigate the efficacy and safety of the long-acting altrenogest injection (NV Readyserve® injection) for horses. DESIGN: A single-dose pharmacokinetic (PK) study was conducted. The in vivo efficacy study was a blinded, repeated measures design evaluating behaviour scores. The safety study was a non-blinded, controlled, parallel-group, randomised-block design as per the VICH protocol. METHODS: In the PK study, serial blood samples were obtained for analysis of plasma altrenogest for 150 h following the injection and a non-compartmental PK analysis was performed. For the efficacy study, 12 mares in oestrus were treated; they were monitored daily for 10 days for signs of oestrus during teasing and given a behaviour score that was compared with pretreatment scores. A standard safety study was conducted at 1-, 3- and 5-fold the recommended dosage for 84 days. Physical, haematological and biochemical examinations were performed. RESULTS: Mean plasma altrenogest concentrations were greater than ≈0.5 ng/mL for 148 h following administration. Oestrous behaviour was suppressed in all mares within 24 h of administration. Two mares returned to oestrus by day 6 and the rest on days 7-10. In the safety study there were no significant differences in the physical and haematological examinations, but minor biochemical changes in muscle enzymes. There was a low incidence of injection site reactions following the 3- and 5-fold dose, predominantly for pectoral injections. CONCLUSION: These studies support the efficacy and safety of a single dose of Readyserve® injection for the suppression of the signs of oestrus in mares for 5-7 days.

Trenbolone Improves Cardiometabolic Risk Factors and Myocardial Tolerance to Ischemia-Reperfusion in Male Rats With Testosterone-Deficient Metabolic Syndrome.

The increasing prevalence of obesity adds another dimension to the pathophysiology of testosterone (TEST) deficiency (TD) and potentially impairs the therapeutic efficacy of classical TEST replacement therapy. We investigated the therapeutic effects of selective androgen receptor modulation with trenbolone (TREN) in a model of TD with the metabolic syndrome (MetS). Male Wistar rats (n=50) were fed either a control standard rat chow (CTRL) or a high-fat/high-sucrose (HF/HS) diet. After 8 weeks of feeding, rats underwent sham surgery or an orchiectomy (ORX). Alzet miniosmotic pumps containing either vehicle, 2-mg/kg·d TEST or 2-mg/kg·d TREN were implanted in HF/HS+ORX rats. Body composition, fat distribution, lipid profile, and insulin sensitivity were assessed. Infarct size was quantified to assess myocardial damage after in vivo ischaemia reperfusion, before cardiac and prostate histology was performed. The HF/HS+ORX animals had increased sc and visceral adiposity; circulating triglycerides, cholesterol, and insulin; and myocardial damage, with low circulating TEST compared with CTRLs. Both TEST and TREN protected HF/HS+ORX animals against sc fat accumulation, hypercholesterolaemia, and myocardial damage. However, only TREN protected against visceral fat accumulation, hypertriglyceridaemia, and hyperinsulinaemia and reduced myocardial damage relative to CTRLs. TEST caused widespread cardiac fibrosis and prostate hyperplasia, which were less pronounced with TREN. We propose that TEST replacement therapy may have contraindications for males with TD and obesity-related MetS. TREN treatment may be more effective in restoring androgen status and reducing cardiovascular risk in males with TD and MetS.

Product-to-parent reversion of trenbolone: unrecognized risks for endocrine disruption.

Trenbolone acetate (TBA) is a high-value steroidal growth promoter often administered to beef cattle, whose metabolites are potent endocrine-disrupting compounds. We performed laboratory and field phototransformation experiments to assess the fate of TBA metabolites and their photoproducts. Unexpectedly, we observed that the rapid photohydration of TBA metabolites is reversible under conditions representative of those in surface waters (pH 7, 25°C). This product-to-parent reversion mechanism results in diurnal cycling and substantial regeneration of TBA metabolites at rates that are strongly temperature- and pH-dependent. Photoproducts can also react to produce structural analogs of TBA metabolites. These reactions also occur in structurally similar steroids, including human pharmaceuticals, which suggests that predictive fate models and regulatory risk assessment paradigms must account for transformation products of high-risk environmental contaminants such as endocrine-disrupting steroids.

New onset diabetes associated with bovine growth hormone and testosterone abuse in a young body builder.

CASE: A 33-year-old male presented to the emergency department with complaints of polydipsia, polyuria, nausea, headaches, blurry vision and malaise. Lab work revealed a serum glucose level of 1166 mg/dl (64.8 mmol/L). The patient admitted to completing a cycle of androgenic anabolic steroids (AASs) for bodybuilding. His regimen consisted of supraphysiologic intramuscular injections of a bovine growth hormone, trenbolone acetate and testosterone. The patient received intravenous fluids and insulin to restore metabolic balance. Previously healthy with a non-contributory family history, he was diagnosed with new onset diabetes. DISCUSSION: It has been demonstrated that AAS use, specifically growth hormone, can affect glucose homeostasis through increasing cellular insulin resistance and reducing glucose uptake. Excess growth hormone has been shown to cause symptoms of acromegaly which predisposes up to 40% of patients to diabetes. As trenbolone acetate is not indicated for human use and athletes are known to use supraphysiologic doses of this underground, performance enhancing drug, the correlation of the timing of events and the use of this veterinary growth hormone likely exacerbated an underlying condition or caused this new onset diabetes. CONCLUSION: We report a case of a young bodybuilder with no significant past medical history who was diagnosed with new onset diabetes associated with supraphysiologic self-injections of the bovine growth hormone, trenbolone acetate, combined with testosterone. AAS have the potential to induce or exacerbate diabetic conditions due to decreased glucose tolerance and increased insulin resistance.

Concentrations of altrenogest in plasma of mares and foals and in allantoic and amniotic fluid at parturition.

Treatment with the progestin altrenogest is widely used in pregnant mares. The fact that foals born from healthy mares treated with altrenogest until term suffered from neonatal problems raises the question of direct effects of altrenogest on vital functions in the neonate. We have therefore investigated altrenogest concentrations in maternal and neonatal blood plasma and in fetal fluids. Pregnant mares were treated with altrenogest orally once daily (0,088 mg/kg bodyweight, n = 7) or left untreated (n = 8) from 280 d of gestation until foaling. Altrenogest concentration was determined in plasma of the mares, their foals and in amniotic and allantoic fluid. The concentration of altrenogest in plasma from treated mares (2.6 +/- 1.0 ng/mL) was significantly lower than in plasma from their foals immediately after birth (5.6 +/- 1.9 ng/mL; p < 0.05), but was significantly higher than in their fetal fluids (amniotic fluid: 0.4 +/- 0.1 ng/mL; p < 0.05; allantoic fluid: 3.0 +/- 1.5 ng/mL). Altrenogest was undetectable in maternal and fetal plasma and fetal fluids of control pregnancies at all times. Altrenogest concentration in plasma of foals from treated mares was strongly correlated to the altrenogest concentration in plasma of their dams (r = 0.938, p < 0.001) and in amniotic (r = 0.886, p < 0.001) and allantoic fluid (r = 0.562, p < 0.05). A significant decrease in altrenogest concentration between the time periods 0-15 min, 30-120 min, and 180-360 min after parturition was seen in the plasma from foals born to altrenogest-treated mares. In conclusion, our data demonstrate that altrenogest reaches the equine fetus at high concentrations.

Real-time PCR-based prediction of gonad phenotype in medaka.

An important endpoint in aquatic bioassays for potential endocrine disrupting chemicals (EDCs) is the gonadal phenotype of exposed fish, with special interest in intersex and sex-reversed individuals. Traditionally, the assessment of gonad phenotype is done via histology, which involves specialized and time-consuming techniques. The method detailed here increases the efficiency of the analysis by first determining the relative expression of four genes involved in gonad development/maintenance in Japanese medaka (Oryzias latipes), and then by using principal component analysis, assigning a phenotype to each gonad based upon the gene expression data. The gonad phenotype and the sexual genotype, which can be determined in medaka, can then be compared to assess potential adverse effects of exposure to endocrine disrupting chemicals.

Induction of micronuclei in V79 cells by the anabolic doping steroids tetrahydrogestrinone and trenbolone.

The synthetic steroid tetrahydrogestrinone is a new "designer drug" and was recently detected to be illegally used in sports. It is chemically closely related to trenbolone that is known as an animal growth promoter. The potencies of trenbolone, tetrahydrogestrinone and testosterone to induce micronuclei in V79 cells in vitro were determined. CREST analysis was employed to differentiate between aneugenic or clastogenic mechanisms. Cytotoxicity and an influence on the cell cycle were assessed in parallel. Incubations with testosterone, at concentrations between 3 and 300 microM, failed to induce micronuclei. By contrast, tetrahydrogestrinone and trenbolone increased the rate of micronuclei significantly, up to a doubling of the micronuclei rate of untreated controls. Tetrahydrogestrinone and trenbolone displayed a bell-shaped dose-response curve, with maximal effects observed at 3 and 30 microM, respectively. The micronuclei induced by tetrahydrogestrinone and trenbolone were predominantly kinetochor (CREST) positive, pointing to an aneugenic mode of action. This may be related to the specific structure of both molecules with a system of activated double bonds. As the genotoxic effect of tetrahydrogestrinone at a chromosomal level appears at a low concentration range, it cannot be ruled out that tetrahydrogestrinone presents a genotoxic hazard on a chromosomal level under conditions of its current misuse in sports.

Effect of prolonged use of altrenogest on behaviour in mares.

Oral administration of altrenogest for oestrus suppression in competition horses is believed to be widespread in some equestrian disciplines, and can be administered continuously for several months during a competition season. To examine whether altrenogest has any anabolic or other potential performance enhancing properties that may give a horse an unfair advantage, we examined the effect of oral altrenogest (0.044 mg/kg), given daily for a period of eight weeks, on social hierarchy, activity budget, body-mass and body condition score of 12 sedentary mares. It was concluded that prolonged oral administration of altrenogest at recommended dose rate to sedentary mares had no effect on dominance hierarchies, body-mass or condition score.

Effect of prolonged use of altrenogest on behaviour in mares.

Oral administration of altrenogest for oestrus suppression in competition horses is believed to be widespread in some equestrian disciplines, and can be administered continuously for several months during a competition season. To examine whether altrenogest has any anabolic or other potential performance enhancing properties that may give a horse an unfair advantage, we examined the effect of oral altrenogest (0.044 mg/kg), given daily for a period of eight weeks, on social hierarchy, activity budget, body-mass and body condition score of 12 sedentary mares. We concluded that prolonged oral administration of altrenogest at recommended dose rates to sedentary mares resulted in no effect on dominance hierarchies, body mass or condition score.

Effects of growth implants on consumer perceptions of meat tenderness in beef steers.

Anabolic steroid implants are routinely used to increase growth performance and profitability; however, there are concerns that the use of implants, particularly those containing trenbolone acetate, may have detrimental effects on carcass quality and beef tenderness. Thus, the objectives of the current study were to determine the effects of various commonly used implant regimens on shear force values, sensory properties, and consumer satisfaction of beef top loin steaks from cattle of Bos indicus influence. Cattle were supplied by producers that agreed to provide sire and dam information in exchange for carcass and sensory data. Steers (n = 2,748) were assigned randomly to one of three implant treatments (12/sire; four steers from each sire were placed into each treatment group): 1) unimplanted controls (n = 1,368); 2) Synovex-S followed by another Synovex-S (n = 660); or 3) Synovex-S followed by Revalor-S (n = 720). Steaks sampled after 3, 7, and 14 d of aging indicated that unimplanted cattle had lower (P < 0.05) Warner-Bratzler Shear force values than those from implanted animals. No differences (P > 0.05) in shear force values were found between the two treatments or the control groups for steaks sampled following a 21-d aging period. Steaks from implanted animals sampled after 3, 7, and 14 d aging were rated lower (P < 0.05) for initial and sustained trained sensory panel tenderness scores. Consumers failed to detect any differences in steak samples related to implant treatment after 7 and 14 d of aging. Consumer education level and family income did not affect overall acceptability (P > 0.10 and 0.18, respectively) or tenderness acceptability (P > 0.11 and 0.68, respectively); however, consumers with postgraduate degrees recorded lower (P < 0.05) overall quality, beef flavor, juiciness, and tenderness scores than consumers in all other education classifications. Additionally, family income had no effect on overall quality (P > 0.21), beef flavor (P > 0.28), juiciness (P > 0.58), or tenderness (P > 0.45) scores. Results indicate that using a moderate implant program in Bos indicus-influenced cattle has no detrimental effects on beef tenderness and consumer acceptability.

Feedlot performance of steers treated concurrently with ceftiofur crystalline-free acid subcutaneously in the posterior aspect of the ear and a growth-promoting implant.

Ceftiofur crystalline-free acid sterile suspension (CCFA-SS), a long-acting formulation of ceftiofur formulated for subcutaneous injection in the middle third of the posterior aspect of the ear, is being developed for the control and treatment of bovine respiratory disease. A study was designed to evaluate average daily gain (ADG) and feed efficiency (FE) for cattle through 140 days in the feedlot after CCFA-SS was administered concurrently in the same ear with a growth-promoting implant. On Day 0, steers (n = 207) averaging 189 kg in weight were randomly assigned to the following treatments: Revalor -S implant (120 mg trenbolone acetate and 24 mg estradiol per implant; Hoechst-Roussel Agri-Vet Company) (n = 64); CCFA-SS at 6.6 mg ceftiofur equivalents/kg and a Revalor -S implant (n = 64); untreated control (no CCFA-SS or implant) (n = 63); or CCFA-SS only (n = 16). On Day 56, an Implus-S implant (200 mg progesterone USP plus 20 mg estradiol benzoate; Pharmacia and Upjohn Animal Health) was administered to all cattle. Tolerance of administration of all materials was observed visually and by palpation of the treated ears. Average daily gain and FE from Day 0 through Day 56 were significantly (P <.001) better for steers of both groups with an implanted growth-promotant than for untreated controls. From Day 0 through Day 140, ADG was significantly (P <.05) better for cattle given an implant or an implant plus CCFA-SS than for untreated controls and FE was significantly (P <.05) better for cattle given an implant plus CCFA-SS than for controls. Mild or moderate, transient swelling of the treated ear was observed in two cattle (CCFA-SS plus implant) on Day 52. On Day 56, 88 % of cattle treated with CCFA-SS, 84 % of the cattle treated with an implant plus CCFA-SS, and 100 % of cattle in other groups were normal. Administration of CCFA-SS in the middle third of the posterior aspect of the ear at the same time as growth-promoting implants did not affect performance of cattle in the feedlot and was well tolerated by the animals.

Quantitative assessment of foetal exposure to trenbolone acetate, zeranol and melengestrol acetate, following maternal dosing in rabbits.

1. Residues of commonly used growth-promoting agents found in animal meat can be hormonally active and they have been implicated as possible endocrine disruptors in man. Although these compounds could be potentially detrimental to the developing foetus, it is not clear whether and to what extent they pass through placental barrier. 2. This issue was addressed using the rabbit as an animal model. Pregnant rabbits were treated with trenbolone acetate, zeranol or melengestrol acetate beginning at gestation day 14. Levels of active substances in plasma were screened by means of specific ELISA systems. The residues of parent compounds and their metabolites were quantified in maternal and foetal tissues on gestation day 27 using validated, sensitive HPLC/ELISA methods. 3. All three compounds crossed the placental barrier and were detectable in foetal tissues. The extent of tissue concentration varied depending on the compound and tissue analysed. Gender differences were observed in some instances.

[Side-effects and doping control of anabolic steroids]. / Bivirkninger og dopingkontrol ved brug af anabole steroider.

We present a case of a 26 year-old male body-builder who had used the anabolic steroids (AS) trenbolone and stanozolol for 10 weeks. After four weeks creatine-kinase (CK) including CK-B (cardiac specific isoenzymes) levels were elevated five to ten times. He was without cardiac symptoms and electrocardiography was normal. Three weeks after he stopped using AS, CK and CK-B levels had normalized. One of the drugs, stanozolol could not be detected by the doping-control, maybe because of a fast excretion time. Sale, use and delivery of AS is prohibited in Denmark, but is used in spite of the prohibition. It is important to spread knowledge about the risks of AS among the users.

Effect of prenatal trenbolone acetate treatment on lamb performance and carcass characteristics.

Forty-three pregnant Dorset and Dorset crossbred ewes were assigned randomly to a control group or implanted with either 300 mg trenbolone acetate (Low TBA) or 1,200 mg trenbolone acetate (High TBA) between d 40 and 60 of gestation. Adjusted weaning weights for ewe lambs were 23.3% less (P less than .10) with vs without TBA treatments. Postweaning ADG of ewe lambs was lower (P less than .05) but ADG of ram lambs was greater (P less than .05) for high TBA vs low TBA. Ewe lambs receiving high TBA had 19% less (P less than .05) gain per unit of feed than those receiving low TBA. Days on test for ewe lambs was greater (P less than .05) due to TBA treatment and for high TBA vs low TBA. Days on test for ram lambs was decreased (P less than .05) due to high TBA compared to low TBA. Subcutaneous fat over the ribeye and lower rib were greater (P less than .05) for high-TBA ewe lambs vs low-TBA ewe lambs. Percentage kidney and pelvic fat of ewe lambs was lower (P less than .05) due to TBA treatments. Ribeye area per unit of carcass weight was lower (P less than .05) in high-TBA ewe lambs vs low-TBA ewe lambs. Yield grade of ewe lambs was lower (P less than .05) for low TBA vs high TBA. Prenatal trenbolone acetate treatment of ewe lambs did not improve their subsequent postnatal growth performance and carcass traits. In addition, TBA implantation of the pregnant ewe produced dystocia and less milk production, as evidenced by the need for more lambs to be grafted.

Assay for trenbolone and its metabolite 17 alpha-trenbolone in bovine urine based on immunoaffinity chromatographic clean-up and off-line high-performance liquid chromatography-thin-layer chromatography.

An high-performance liquid chromatography (HPLC)-thin-layer chromatography (TLC) method was developed to detect the illegal use of the xenobiotic growth promotor Trenbolone acetate (TBA). Very effective clean-up of bovine urine was achieved by immunoaffinity chromatography (IAC). The active form of TBA, the steroid 17 beta-Trenbolone (17 beta-TB), as well as its major metabolite 17 alpha-Trenbolone (17 alpha-TB), were assayed simultaneously with HPLC and on-line UV detection. The fraction containing 17 alpha-TB and 17 beta-TB (TB-fraction) was collected, and for confirmation 17 beta- and 17 alpha-TB were subsequently separated and identified by TLC. The limit of detection by on-line HPLC-UV (350 nm) was 1-2 micrograms TB/l. Off-line TLC detection was even more sensitive, 0.5 microgram 17 beta- or 17 alpha-TB/1. The assay was validated by investigating urine samples from veal calves implanted with TBA. The presence of 17 beta- and 17 alpha-TB was clearly demonstrated. A survey of the illegal use of TBA in cattle was performed by applying the assay to urine obtained at slaughter. No residues of TBA or its metabolites were found in any of the 144 random samples from the Dutch public health surveillance programme.

The effect of altrenogest, an oral progestin, on hematologic and biochemical parameters in mares.

Twenty mares were assigned to 1 of 4 groups: no altrenogest; altrenogest at 0.044 mg/kg BW; altrenogest at 0.132 mg/kg BW; or altrenogest at 0.220 mg/kg BW. Treatment was administered daily for 86 days. No signs of illness attributable to feeding altrenogest were observed during the trial. Treatment had no effect (P greater than .05) on the following parameters: WBC, differential WBC, platelet number, creatinine, LDH, CPK, total bilirubin, cholesterol, globulin, BSP, and erythrocyte sedimentation rate. When comparing values over time with pretreatment means or among treatment groups, there were differences (P less than .05) in RBC, PCV, Hb, ALT, PT, PTT, P, Na, TP, BUN, Cl and glucose. However, these changes remained within established normal ranges and also occurred in mares in the control group. There was no treatment by time interaction for any of these parameters. Treatment differences (P less than .05) were observed for K, Ca, alkaline phosphatase and AST during the course of the trial. However, only occasional values of these parameters were outside the established ranges. They were only slightly elevated and tended to be either sporadic or also occurred in control mares. Few of the observed changes could be attributed to the feeding of altrenogest.

Neuromuscular studies of turkeys with trienbolone acetate-induced and naturally occurring "leg weakness".

1. In vivo sciatic nerve gastrocnemius muscle preparations were made from 14- to 18-week-old normal turkeys and from those with naturally occurring or trienbolone acetate (TA)-induced "leg weakness". 2. Preparations from leg weakness cases displayed neither accelerated muscle fatigue nor decreased nerve conduction velocities as compared with control preparations. 3. The muscles of the TA-treated turkeys but not of those with naturally occurring leg weakness were hyperexcitable to those of controls. 4. Post-tetanic potentiation of preparations from both naturally occurring and experimentally-induced cases of leg weakness was less than that of control preparations, both before and after partial neuromuscular blockade, significantly so at the lower pulse frequencies used. 5. It is tentatively suggested that the last finding might be of significance in explaining the clinical signs of leg weakness and might be associated with a disordered calcium metabolism.