Separation of Structurally Similar Anabolic Steroids as Cation Adducts in FAIMS-MS / Michael S Wei, Robin H J Kemperman, Michelle A Palumbo, Richard A Yost. - (Journal of the American Society for Mass Spectrometry 31 (2020) 2 (5 February); p. 355-365)

• PMID: 32031405.
• DOI: 10.1021/jasms.9b00127

Abstract

Novel synthetic anabolic androgenic steroids have been developed not only to dodge current antidoping tests at the professional sports level, but also for consumption by noncompetitive bodybuilders. These novel anabolic steroids are commonly referred to as "designer steroids" and pose a significant risk to users because of the lack of testing for toxicity and safety in animals or humans. Manufacturers of designer steroids dodge regulation by distributing them as nutritional or dietary supplements. Improving the throughput and accuracy of screening tests would help regulators to stay on top of illicit anabolic steroids. High-field asymmetric-waveform ion mobility spectrometry (FAIMS) utilizes an alternating asymmetric electric field to separate ions by their different mobilities at high- and low-fields as they travel through the separation space. When coupled to mass spectrometry (MS), FAIMS enhances the separation of analytes from other interfering compounds with little to no increase in analysis time. Here we investigate the effects of adding various cation species to sample solutions for the separation of structurally similar or isomeric anabolic androgenic steroids. FAIMS-MS spectra for these cation-modified samples show an increased number of compensation field (CF) peaks, some of which are confirmed to be unique for one steroid isomer over another. The CF peaks observed upon addition of cation species correspond to both monomer steroid-cation adduct ions and larger multimer ion complexes. Notably, the number of CF peaks and their CF shifts do not appear to have a straightforward relationship with cation size or electronegativity. Future directions aim at investigating the structures for these analyte-cation adduct ions for building a predictive model for their FAIMS separations.

Serum insulin-like factor 3 levels are reduced in former androgen users suggesting impaired Leydig cell capacity / Jon Jarløv Rasmussen, Jakob Albrethsen, Mikkel Nicklas Frandsen, Niels Jørgensen, Anders Juul, Caroline Kistorp. - (The Journal of Clinical Endocrinology & Metabolism (2021) 9 March)

• PMID: 33693710
• DOI: 10.1210/clinem/dgab129

Abstract

Background: Illicit use of anabolic androgenic steroids (AAS) is frequently observed in men and is associated with subsequent testosterone deficiency although the long-term effect on gonadal function is still unclear. Serum insulin-like factor 3 (INSL3) has been suggested to be a superior biomarker of Leydig cell secretory capacity compared to testosterone. The objective of this study was to investigate serum INSL3 concentrations in AAS users.

Methods: This community-based cross-sectional study included men aged 18 - 50 years, involved in recreational strength training and allocated to one of three groups: never-AAS users as controls (n=44), current (n=46) or former AAS users (n=42) with an average duration since AAS cessation of 32 (23;45) months.

Results: Serum INSL3 was lower in current AAS users and former AAS users than in controls, median (IQR), 0.04 (ND - 0.07) and 0.39 (0.24 - 0.62) versus 0.59 (0.45 - 0.72) µg/L, P<0.001. Former AAS users exhibited lower serum INSL3 levels than controls in a multivariate linear regression even after adjusting for serum total testosterone and other relevant confounders, (B) (95%CI), -0.16 (-0.29;-0.04) µg/L, P=0.011. INSL3 and total testosterone were not associated in the model, P=0.821. Longer accumulated AAS duration (log2) was associated with lower serum INSL3 in former AAS users, (B) (95%CI), -0.08 (-0.14;-0.01), P=0.022. Serum INSL3, but not inhibin B or testosterone, was associated with testicular size in a multivariate linear regression, (B) (95%CI); 4.7 (0.5 ; 8.9), P=0.030.

Conclusions: Serum INSL3 is reduced years following AAS cessation in men, independently of testosterone, suggesting persistently impaired Leydig cell capacity.

Serum levels of enclomiphene and zuclomiphene in men with hypogonadism on long-term clomiphene citrate treatment / Sevann Helo, Joseph Mahon, Joseph Ellen, Ron Wiehle, Gregory Fontenot, Kuang Hsu, Paul Feustel, Charles Welliver, Andrew McCullough

• BJUI 119 (2017) 1 (January), 171-176
• PMID: 27511863
• DOI: 10.1111/bju.13625

Abstract

Objectives: To determine the relative concentrations of enclomiphene (ENC) and zuclomiphene (ZUC) isomers in men with hypogonadism on long-term clomiphene citrate (CC) therapy, and to determine whether patient age, body mass index (BMI) or duration of therapy were predictive of relative concentrations of ENC and ZUC.

Patients and methods: Men already receiving CC 25 mg daily therapy for secondary hypogonadism for a minimum of 6 weeks were recruited to have their ENC and ZUC levels assessed. Total testosterone, free testosterone, oestradiol, follicle stimulating hormone (FSH), and luteinizing hormone (LH) before initiation of and while on CC therapy were recorded for all patients. Patient demographics including age, BMI and medical comorbidites were recorded. Serum samples were obtained at the time of enrolment to determine ENC and ZUC concentrations.

Results: A total of 15 men were enrolled in the period from June 2015 to August 2015. The median (range) patient age was 36 (22-70) years, BMI 32.0 (21.1-40.3) kg/m2 and duration of treatment 25.9 (1.7-86.6) months. Baseline median total testosterone, oestradiol and LH levels were 205.0 ng/dL, 17.0 pg/mL and 4.0 mlU/mL, respectively. The post-treatment median total testosterone, oestradiol and LH level increased to 488.0 ng/dL, 34.0 pg/mL and 6.1 mIU/mL, respectively (all P<0.001). The median ENC and ZUC concentrations were 2.2 and 44.0 ng/mL, respectively. After at least 6 weeks of CC therapy, the median ZUC: ENC serum concentration ratio was 20:1. On linear regression analysis. patient age, BMI, duration of treatment and serum testosterone levels were not predictive of ENC or ZUC concentrations.

Conclusions: Long-term CC therapy resulted in a significant alteration of ENC and ZUC concentrations, with ZUC as the predominant isomer. Given the vastly different biochemical and toxicological properties of ENC and ZUC, this study supports the need for the development of a pure selective oestrogen receptor antagonist for the treatment of men with hypogonadism.

Severe Acquired Hypokalemic Paralysis in a Bodybuilder After Self-medication With Triamterene/Hydrochlorothiazide / Nikolaus Pfisterer, Josef Finsterer. - (Clinical Journal of Sport Medicine (2019) 18 November).
- PMID: 31770156.
- DOI: 10.1097/JSM.0000000000000763

Abstract

Background:
Severe hypokalemia with severe neurological impairment and electrocardiogram (ECG) abnormalities due to the misuse of triamterene/hydrochlorothiazide (HCTZ) in a bodybuilder has not yet been reported.

Case report:
A 22-year-old bodybuilder developed acute generalized muscle cramps, sensory disturbance of the distal lower and upper limbs, quadriparesis, and urinary retention. These abnormalities were attributed to severe hypokalemia of 1.8 mmol/L (normal range 3.4-4.5 mmol/L) due to misuse of triamterene/HCTZ together with fluid restriction. He was cardiologically asymptomatic, but ECG revealed a corrected QT (QTc) interval of 625 ms. On intravenous application of fluids along with intravenous and oral substitution of potassium, his condition rapidly improved, such that the sensory disturbances, quadriparesis, and bladder dysfunction completely resolved within 2 days after admission.

Conclusions:
Self-medication with diuretics along with fluid restriction may result in severe hypokalemia, paralysis, and ECG abnormalities. Those responsible for the management of bodybuilding studios and competitions must be aware of the potential severe health threats caused by self-medication with diuretics and anabolic steroids. Although triamterene is potassium-sparing, it may enhance the potassium-lowering effect of HCTZ.

Severe Cardiac and Metabolic Pathology Induced by Steroid Abuse in a Young Individual / Adrian Tirla, Cosmin Mihai Vesa, Simona Cavalu. - (Diagnostics 11 (2021) 8 (21 July); p. 1-13)

• PMID: 34441248
• PMCID: PMC8394374
• DOI: 10.3390/diagnostics11081313

Abstract

Androgenic-Anabolic Steroids (AAS) abuse is known to play an important role in causing the systemic inflammatory response and multiple-organ dysfunction in healthy individuals. Although many of the undesirable effects of steroid abuse have been reported, at present, little is known about the effect of anabolic supplements and the correlation between cardiac and metabolic pathology. This paper presents a case of a 25 year old patient with a complex medical history after 6 months of steroid administration. Myocardial infraction, dyslipidemia, obesity, hyperuricemia, secondary diabetes, and chronic renal disease were identified after clinical and para-clinical examinations. The particularities of this case were interpreted in the context of a literature review, highlighting the effect of multi-organ damage as a result of the uncontrolled use of anabolic steroid supplements.

Severe Persistent Jaundice after the Abuse of an Anabolic Androgenic Steroid Analogue / Hyun Gil Goh, Yoo Jin Lee, Tae Hyung Kim. - (Korean Journal of Gastroenterology 76 (2020) 3 (25 September); p. 167-170)

• PMID: 32969366
• DOI: 10.4166/kjg.2020.76.3.167

Abstract

Hepatic disorders with prominent cholestasis can be caused by a range of conditions, and anabolic androgenic steroids have been considered a cause of protracted cholestasis. A 29-year-old man who had taken an anabolic androgenic steroid analogue for 2 months visited the hospital complaining of jaundice and indigestion. After stopping the medication, the hyperbilirubinemia tended to decrease, but a transiently elevated aminotransferase level was observed. The endogenous testosterone level also decreased initially but recovered soon after. The liver function profiles were normalized after 2 months of conservative management. This case emphasizes that close drug history taking, including anabolic steroids, is important for identifying the cause of unexplained persistent jaundice.

Keywords: Anabolic agents; Chemical and drug induced liver injury; Jaundice; Steroids; Substance abuse; oral.

Severity of anabolic steroid dependence, executive function, and personality traits in substance use disorder patients in Norway / Morgan Scarth, Ingrid A. Havnes, Marie L. Jørstad, Jim McVeigh, Marie Claire Van Hout, Lars T. Westlye, Svenn Torgersen, Astrid Bjørnebekk

• Drug and Alcohol Dependence 231 (1 February 2022), 109275
• PMID: 35030506
• DOI: 10.1016/j.drugalcdep.2022.109275

Abstract

Introduction: Anabolic androgenic steroids (AAS), including testosterone and synthetic derivatives, are typically used to increase muscle mass. Many users develop a dependence on these substances, contributing to worsened physical and mental health outcomes. Aspects of personality and executive dysfunction may represent underlying vulnerabilities for developing dependence.

Objective: To identify levels of AAS dependence within substance use disorder (SUD) treatment patients and assess the relationship between dependence severity and personality traits and executive function (EF).

Methods: Data were collected from patients at 38 SUD treatment facilities in Norway. Questionnaires were completed for measures of personality and EF. Measures of symptoms of AAS dependence were used in latent class analysis to identify sub-groups of patients, which were evaluated for association with EF and personality traits, and compared with a group of non-AAS using SUD patients.

Results: Three classes were identified; largely reflecting low, moderate, and high symptoms of dependence. Multinomial regression analyses indicated that moderate and high symptoms were associated with several measures of EF and personality traits, particularly self-monitoring, antagonism, disinhibition, and rigid perfectionism while users with low symptoms exhibited higher capacities for emotional control and shift, and lower negative affectivity, relative to non-AAS using SUD patients. Backward stepwise regressions indicated antagonism, and decreased self-monitoring as key personality and cognitive characteristics of SUD patients with severe AAS dependence.

Conclusion: Our findings indicate that specific executive dysfunctions and personality features, particularly those associated with poor emotional control, reduced empathy, and impulsivity are associated with more severe AAS dependence in the SUD population.

Keywords: Anabolic androgenic steroids; Executive function; Latent class analysis; Personality; Substance use disorder.

Short term impact of Tribulus terrestris intake on doping control analysis of endogenous steroids / Christophe Saudan, Norbert Baume, Caroline Emery, Emmanuel Strahm, Martial Saugy. - (Forensic Science International 178 (2008) 1 (10 June); p. e7-e10)

• PMID: 18282674
• DOI: 10.1016/j.forsciint.2008.01.003

Abstract

Tribulus terrestris is a nutritional supplement highly debated regarding its physiological and actual effects on the organism. The main claimed effect is an increase of testosterone anabolic and androgenic action through the activation of endogenous testosterone production. Even if this biological pathway is not entirely proven, T. terrestris is regularly used by athletes. Recently, the analysis of two female urine samples by GC/C/IRMS (gas chromatography/combustion/isotope-ratio-mass-spectrometry) conclusively revealed the administration of exogenous testosterone or its precursors, even if the testosterone glucuronide/epitestosterone glucuronide (T/E) ratio and steroid marker concentrations were below the cut-off values defined by World Anti-Doping Agency (WADA). To argue against this adverse analytical finding, the athletes recognized having used T. terrestris in their diet. In order to test this hypothesis, two female volunteers ingested 500 mg of T. terrestris, three times a day and for two consecutive days. All spot urines were collected during 48 h after the first intake. The (13)C/(12)C ratio of ketosteroids was determined by GC/C/IRMS, the T/E ratio and DHEA concentrations were measured by GC/MS and LH concentrations by radioimmunoassay. None of these parameters revealed a significant variation or increased above the WADA cut-off limits. Hence, the short-term treatment with T. terrestris showed no impact on the endogenous testosterone metabolism of the two subjects.

Short-term growth hormone treatment does not increase muscle protein synthesis in experienced weight lifters / Kevin E. Yaraheski, Jeffrey J. Zachwieja, Theodore J. Angelopoulos, Dennis M. Bier. - (Journal of Applied Physiology 74 (1993) 6 (June); p. 3073-3076)

• PMID: 8366011
• DOI: 10.1152/jappl.1993.74.6.3073

Abstract

The purpose of this study was to determine whether recombinant human growth hormone (GH) administration enhances muscle protein anabolism in experienced weight lifters. The fractional rate of skeletal muscle protein synthesis and the whole body rate of protein breakdown were determined during a constant intravenous infusion of [13C]leucine in 7 young (23 +/- 2 yr; 86.2 +/- 4.6 kg) healthy experienced male weight lifters before and at the end of 14 days of subcutaneous GH administration (40 microgram.kg-1 x day-1). GH administration increased fasting serum insulin-like growth factor-I (from 224 +/- 20 to 589 +/- 80 ng/ml, P = 0.002) but did not increase the fractional rate of muscle protein synthesis (from 0.034 +/- 0.004 to 0.034 +/- 0.002%/h) or reduce the rate of whole body protein breakdown (from 103 +/- 4 to 108 +/- 5 mumol.kg-1 x h-1). These findings suggest that short-term GH treatment does not increase the rate of muscle protein synthesis or reduce the rate of whole body protein breakdown, metabolic alterations that would promote muscle protein anabolism in experienced weight lifters attempting to further increase muscle mass.

Short-term rhGH increases PIIINP, a biomarker of endothelial dysfunction / Michael R. Graham, Yaodong Gu, P.J. Evans, S.M. Cooper, Bruce Davies, Julien S. Baker. - (International Journal of Advanced Engineering Research and Science 4 (2017) 7 (July); p. 164-173)

• DOI: 10.22161/ijaers.4.7.26

Abstract:

Objectives: In arterial hypertension, amino-terminal pIII procollagen ropeptide of type (PIIINP) is elevated in arterial aneurysm tissue and associated with a poor prognosis following acute myocardial infarction (MI). Recombinant human growth hormone (rhGH) administration attenuates endothelial dysfunction but increases PIIINP. This study was conducted to establish if short-term rhGH administration affects PIIINP, endothelial function and selected cardiovascular disease (CVD) risk factors, in healthy males.

Design: Method: Male subjects (n=48) were randomly assigned into two groups: (1): control group (C) n=24, mean ± SD, age 32 ± 11 years; height 1.8 ± 0.06 metres; (2): rhGH administration group (rhGH) n=24, mean ± SD, age 32 ± 9 years; height 1.8 ± 0.07 metres. Blood pressure (BP), heart rate (HR), arterial pulse wave velocity (APWV), and biochemical indices were investigated.

Results: PIIINP (0.28±0.1 vs. 0.42±0.2, U/ml); Insulin like growth factor-I (159±54 vs. 323±93, ng.mL-1); resting HR (72±14 vs. 78±11, b.p.m.) and rate pressure product (RPP) (90±18 vs. 97±14, bpm x mm.Hg x 10-2) all significantly increased (P<0.05). Total cholesterol (4.7±0.9 vs. 4.4±0.7, mmol.L-1); high sensitivity C-reactive protein (1.77±2.1 vs. 1.29±1.6, mg.L-1); serum homocysteine (13.2±4.0 vs. 11.7±3.1, μmol.L-1) and APWV (9.97±1.38 vs. 9.18±1.6, m.s-1) all significantly decreased (P<0.05).

Conclusion: Paradoxically, there was an improvement in CVD inflammatory markers and APWV; but PIIINP and resting RPP increased. Elevated PIIINP may have a confounding adverse effect on the endothelium, but may also provide clinical prognostic information in monitoring arterial hypertension, left ventricular function in the sub-acute phase following MI and endothelial function in aortic aneurysms.