Testosterone Derivatives Research Articles

Anabolic-androgenic steroid use among bodybuilders in Erbil city

Background and objective: Anabolic-androgenic steroids are synthetic testosterone derivatives with a longer duration of action than physiological androgens. They are abused by bodybuilders because of their potential to enhance muscle strength. Serious medical and psychological complications may be associated with their non-medical use. This study aimed to determine the prevalence of anabolic androgenic steroids among bodybuilders and their awareness of health-related risks. Methods: A cross-sectional study was carried out from the 1st of January 2019 to the end of July 2019 in 18 randomly selected gym centers in Erbil city, Iraq using stratified random sampling. A self-administered questionnaire was distributed to 400 bodybuilders to assess the prevalence and knowledge of using Anabolic-androgenic steroids. Results: The study showed a high prevalence of Anabolic-androgenic steroid consumption (26.3%) among gym users in Erbil city. Anabolic-androgenic steroid use was significantly higher among participants with a longer duration of bodybuilding practice ≥4 years (P = 0.001). Anabolic-androgenic steroid abuse was significantly associated with drinking alcohol, smoking cigarettes, and using growth hormones. Trainers were the commonest source of recommendation. Conclusion: The prevalence of anabolic androgenic steroids abuse is high among bodybuilders in Erbil city. Public health awareness is essential and may help avoid the propagation of the problem. Keywords: Anabolic steroids; Gym users; Bodybuilders; Knowledge, Erbil.

Synthesis and Actions of 5α-Reduced Metabolites of Testosterone in the Nervous System

Neuroactive testosterone metabolites synthesized by enzymes 5α-reductase and 3α/3β-hydroxysteroid oxidoreductase are critical physiological modulators of the central nervous system (CNS) function. Indeed, as discussed in this review, they exert a variety of effects on myelination, brain maturation, neurotransmission, reproductive behavior, and the stress responses. The expression and synthesis of these enzymes as well as the levels of testosterone derivatives exhibit specific localized regional and sex differences in the CNS and are influenced under various physiological and neuropathological conditions. In addition, neuroactive metabolites of testosterone exert a variety of neuroprotective effects in several neuropathological animal models. Therefore, neuroactive testosterone metabolites might represent an interesting potential therapeutic option to treat some neurological disorders.

ANABOLIC-ANDROGENIC STEROIDS ARE ASSOCIATED WITH MAJOR DEPRESSIVE DISORDER AND SUICIDE ATTEMPT: ANALYSIS OF A MULTI-NATIONAL DATABASE

Anabolic-androgenic steroids (AAS), synthetic derivatives of the male sex hormone testosterone, are often abused to enhance athletic performance and physical appearance. Long term use of AAS can be associated with mood destabilizing effects and psychotic behavior. We hypothesized that extended AAS use or abuse is an independent risk factor for body dysmorphic disorder, depression, and suicidality. We investigated AAS abuse and associations with major depressive disorder (MDD), suicide attempt, and body dysmorphic disorder (BDD) by querying TriNetX, a global health research network that provides data on large cohort of patients from multiple centers across the United States.

Boldenone Undecylenate-Mediated Hepatorenal Impairment by Oxidative Damage and Dysregulation of Heat Shock Protein 90 and Androgen Receptors Expressions: Vitamin C Preventive Role.

Boldenone Undecylenate (BLD) is a synthetic derivative of testosterone and a widely used anabolic androgenic steroid. The health risk of BLD use as a pharmaceutical or dietary supplement is still underestimated and under-reported. Vitamin C (VC) has been recognized as an antioxidant with prominent hepatorenal protective effects. This study investigated the possible preventive activity of VC against BLD-induced hepatorenal damage. Forty adult male Wistar rats were classified into five groups: control, vehicle control, VC (orally given 120 mg/kg b. wt./day), BLD (intramuscularly injected 5 mg/kg b. wt./week), and BLD + VC-treated groups. The experiment continued for eight weeks. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. Serum contents of total protein (TP), albumin (ALB), globulin, total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and very-low-density lipoprotein–cholesterol (VLDL-C) were also assayed. Urea, creatinine, and uric acid levels were determined together with sodium and potassium electrolytes measuring. Moreover, oxidative stress indicators including reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and glutathione reductase (GSR) as well as malondialdehyde (MDA) levels were measured in both hepatic and renal tissues. Corresponding histological examination of renal and hepatic tissues was conducted. Besides, immunohistochemical evaluations for androgen receptors protein (AR) and heat shock protein 90 (Hsp 90) expressions were performed. BLD caused significant rises in serum ALT, AST, TP, ALB, TC, TG, LDL-C, VLDL-C, urea, creatinine, uric acid, potassium, and MDA levels. Further, BLD-injected rats showed significant declines in the serum levels of HDL-C, sodium, GSH, GPx, GST, and GSR. Besides, distinct histopathological perturbations were detected in renal and hepatic tissues of BLD-injected rats. AR and Hsp 90 immunoexpression were increased in hepatic and renal tissues. In contrast, VC significantly reversed the BLD-induced hepatorenal damage in co-treated rats but not ameliorated AR protein overexpression. VC could be an efficient preventive supplement for mitigating BLD-induced hepatorenal damage, possibly via controlling oxidative stress events.

HEPATOTOXIC AND CARDIOTOXIC EFFECTS OF TESTOSTERONE ENANTHATE ABUSE ON ADULT MALE ALBINO RATS

Background: Anabolic androgenic steroids (AASs) represent a group of synthetic derivatives of testosterone created to maximize anabolic effects and minimize the androgenic ones. Athletes use anabolic androgenic steroids (AAS) to enhance performance regardless of the health risk they may impact in some persons. Objective: To study the histopathological and biochemical changes that could occur on liver and cardiac muscle after administration of doping dose (supraphysiological dose) of testosterone enanthate (one of AAS). Materials and Methods: One hundred (100) male albino rats were included in the present study and divided into six groups. Group (1a); negative control group, Group (1b) positive control group treated with therapeutic dose of testosterone Enanthate (10 mg /Kg body weight intramuscular / I.M weekly for 12 weeks), group (2) was treated with doping dose of testosterone enanthate (60 mg /Kg body weight/ I.M for 4 weeks), group (3) was treated with treated with doping dose of testosterone enanthate (60 mg /Kg body weight / I.M weekly for 8 weeks), group (4) treated with doping dose of testosterone enanthate (60 mg /Kg body weight / I.M weekly for 12 weeks), group (5a) treated with doping dose of testosterone enanthate (60 mg /Kg body weight/ I.M/ weekly for 12 weeks followed by 2 weeks of treatment discontinuation) , group (5b) treated with doping dose of testosterone enanthate (60 mg /Kg body weight/ I.M weekly for 12 weeks followed by 4 weeks of treatment discontinuation). Blood samples were collected at the start and at the end of study. Liver and cardiac enzymes were measured by usual methods. At the end of study for each group, animals were sacrificed and deeply anesthetized by diethyl ether then specimens from the heart and liver were processed for histological study using light microscope. Results: There was no statistical significant difference in the body weight between negative and positive control group. In contrast, animals administering doping dose of testosterone enanthate for 8 and 12 weeks (group III-IV) showed a significant decrease in the body weight compared with animals of control group and group II (P < 0.05). There was a significant decrease in the body weight of recovery group (group Va and group Vb) when compared with control group (p value < 0.05). In the current study as regard the level of liver biomarkers (ALT-AST) had gradual significant increase all through the experimental periods. There was a significant increase in the level of creatinine kinase (CK) and CK-MB in treated groups versus control in most of experimental groups at P Conclusion: Chronic use of testosterone enanthate in supraphysiological dose (doping dose) has a toxic effect on rat cardiac muscle and liver in the form of alteration of normal histology, hypertrophy and fibrosis at both histological and biochemical levels which may lead to loss of their functions. This effect was partially reversible after cessation of the treatment.

Exploring the Potential of Cytochrome P450 CYP109B1 Catalyzed Regio-and Stereoselective Steroid Hydroxylation.

Cytochrome P450 enzyme CYP109B1 is a versatile biocatalyst exhibiting hydroxylation activities toward various substrates. However, the regio- and stereoselective steroid hydroxylation by CYP109B1 is far less explored. In this study, the oxidizing activity of CYP109B1 is reconstituted by coupling redox pairs from different sources, or by fusing it to the reductase domain of two self-sufficient P450 enzymes P450RhF and P450BM3 to generate the fused enzyme. The recombinant Escherichia coli expressing necessary proteins are individually constructed and compared in steroid hydroxylation. The ferredoxin reductase (Fdr_0978) and ferredoxin (Fdx_1499) from Synechococcus elongates is found to be the best redox pair for CYP109B1, which gives above 99% conversion with 73% 15β selectivity for testosterone. By contrast, the rest ones and the fused enzymes show much less or negligible activity. With the aid of redox pair of Fdr_0978/Fdx_1499, CYP109B1 is used for hydroxylating different steroids. The results show that CYP109B1 displayed good to excellent activity and selectivity toward four testosterone derivatives, giving all 15β-hydroxylated steroids as main products except for 9 (10)-dehydronandrolone, for which the selectivity is shifted to 16β. While for substrates bearing bulky substitutions at C17 position, the activity is essentially lost. Finally, the origin of activity and selectivity for CYP109B1 catalyzed steroid hydroxylation is revealed by computational analysis, thus providing theoretical basis for directed evolution to further improve its catalytic properties.

Medicinal Use of Testosterone and Related Steroids Revisited.

Testosterone derivatives and related compounds (such as anabolic-androgenic steroids—AAS) are frequently misused by athletes (both professional and amateur) wishing to promote muscle development and strength or to cover AAS misuse. Even though these agents are vastly regarded as abusive material, they have important pharmacological activities that cannot be easily replaced by other drugs and have therapeutic potential in a range of conditions (e.g., wasting syndromes, severe burns, muscle and bone injuries, anemia, hereditary angioedema). Testosterone and related steroids have been in some countries treated as controlled substances, which may affect the availability of these agents for patients who need them for therapeutic reasons in a given country. Although these agents are currently regarded as rather older generation drugs and their use may lead to serious side-effects, they still have medicinal value as androgenic, anabolic, and even anti-androgenic agents. This review summarizes and revisits the medicinal use of compounds based on the structure and biological activity of testosterone, with examples of specific compounds. Additionally, some of the newer androgenic-anabolic compounds are discussed such as selective androgen receptor modulators, the efficacy/adverse-effect profiles of which have not been sufficiently established and which may pose a greater risk than conventional androgenic-anabolic agents.

Synthesis of Two Testosterone Derivatives and their Theoretical Evaluation as Serotonin Reuptake Transporter Inhibitors

This investigation aimed to synthesize two testosterone derivatives (4 and 7) from either testosterone 3-oxime or testosterone 3-(O-carboxymethyl)oxime. The chemical structure of the compounds was determined using nuclear magnetic resonance spectra. Besides, testosterone derivatives' theoretical activity on serotonin transporter (5i6z protein) was evaluated using fluoxetine as a control in a Docking model. The results showed a higher interaction of both compounds 4 and 7 with a 5i6z protein surface compared with fluoxetine. In conclusion, it's noteworthy that reagents used in this investigation no require special conditions. Also, the theoretical study showed that either compounds 4 or 7 could be good serotonin transporter inhibitors.

Analysis of seized stanozolol formulations in South Brazil by liquid chromatography coupled to quadrupole time-of-flight-mass spectrometry

Anabolic-androgenic steroids (AAS) are synthetic derivatives of testosterone which are used medically for several diseases. However, misuse is commonly observed by athletes to promote enhancement of strength and performance. AAS are frequently obtained through online black markets from clandestine drug manufacturing laboratories, without any quality standards, being potentially dangerous for users. The purpose of this work was the development and application of a fast and simple procedure for the quantitation of stanozolol by liquid chromatography coupled to quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) in tablets packs seized in Rio Grande do Sul state, Brazil. The samples of stanozolol were separated considering its dosage form. The internal standard (methyltestosterone) was added to the aliquots of the samples, dissolved in methanol and 5μL were injected into the analytical system. The newly developed method has been validated for lower limit of quantitation (LLOQ), linearity, accuracy, precision and selectivity. The LLOQ was 0.1 µg/mL. The developed method was successfully applied to 31 samples seized by the Secretaria da Receita Federal do Brasil (a Brazilian federal revenue service agency). According to the results, 90.3% of the suspected medicines (n=31) were adulterated, and 65% exhibited higher concentrations of stanozolol than those indicated in the label. This work successfully established a new method for quantification of stanozolol using LC-QTOF-MS. This method aims at contributing to the identification and quantification of this anabolic androgenic steroid frequently seized by federal inspection agencies.

Nandrolone Decanoate: Use, Abuse and Side Effects.

Background and Objectives: Androgens play a significant role in the development of male reproductive organs. The clinical use of synthetic testosterone derivatives, such as nandrolone, is focused on maximizing the anabolic effects and minimizing the androgenic ones. Class II anabolic androgenic steroids (AAS), including nandrolone, are rapidly becoming a widespread group of drugs used both clinically and illicitly. The illicit use of AAS is diffused among adolescent and bodybuilders because of their anabolic proprieties and their capacity to increase tolerance to exercise. This systematic review aims to focus on side effects related to illicit AAS abuse, evaluating the scientific literature in order to underline the most frequent side effects on AAS abusers’ bodies. Materials and Methods: A systematic review of the scientific literature was performed using the PubMed database and the keywords “nandrolone decanoate”. The inclusion criteria for articles or abstracts were English language and the presence of the following words: “abuse” or “adverse effects”. After applying the exclusion and inclusion criteria, from a total of 766 articles, only 148 were considered eligible for the study. Results: The most reported adverse effects (found in more than 5% of the studies) were endocrine effects (18 studies, 42%), such as virilization, gynecomastia, hormonal disorders, dyslipidemia, genital alterations, and infertility; cardiovascular dysfunctions (six studies, 14%) such as vascular damage, coagulation disorders, and arteriosus hypertension; skin disorders (five studies, 12%) such as pricking, acne, and skin spots; psychiatric and mood disorders (four studies, 9%) such as aggressiveness, sleep disorders and anxiety; musculoskeletal disorders (two studies, 5%), excretory disorders (two studies, 5%), and gastrointestinal disorders (two studies, 5%). Conclusions: Based on the result of our study, the most common adverse effects secondary to the abuse of nandrolone decanoate (ND) involve the endocrine, cardiovascular, skin, and psychiatric systems. These data could prove useful to healthcare professionals in both sports and clinical settings.

Kinetics of Electrophilic Fluorination of Steroids and Epimerisation of Fluorosteroids.

Fluorinated steroids, which are synthesised by electrophilic fluorination, form a significant proportion of marketed pharmaceuticals. To gain quantitative information on fluorination at the 6‐position of steroids, kinetics studies were conducted on enol ester derivatives of progesterone, testosterone, cholestenone and hydrocortisone with a series of electrophilic N−F reagents. The stereoselectivities of fluorination reactions of progesterone enol acetate and the kinetic effects of additives, including methanol and water, were investigated. The kinetics of epimerisation of 6β‐fluoroprogesterone to the more pharmacologically active 6α‐fluoroprogesterone isomer in HCl/acetic acid solutions are detailed.

The efficient Δ1-dehydrogenation of a wide spectrum of 3-ketosteroids in a broad pH range by 3-ketosteroid dehydrogenase from Sterolibacterium denitrificans

Cholest-4-en-3-one Δ1-dehydrogenase (AcmB) from Sterolibacterium denitrificans, a key enzyme of the central degradation pathway of cholesterol, is a protein catalyzing Δ1-dehydrogenation of a wide range of 3-ketosteroids. In this study, we demonstrate the application of AcmB in the synthesis of 1-dehydro-3-ketosteroids and investigate the influence of reaction conditions on the catalytic performance of the enzyme. The recombinant AcmB expressed in E. coli BL21(DE3)Magic exhibits a broad pH optimum and pH stability in the range of 6.5 to 9.0. The activity-based pH optimum of AcmB reaction depends on the type of electron acceptor (2,6-dichloroindophenol - DCPIP, phenazine methosulfate - PMS or potassium hexacyanoferrate - K3[Fe(CN)6]) used in the biocatalytic process yielding the best kinetic properties for the reaction with a DCPIP/PMS mixture (kcat/Km = 1.4·105 s-1·M-1 at pH 9.0) followed by DCPIP (kcat/Km = 1.0·105 s-1·M-1 at pH = 6.5) and K3[Fe(CN)6] (kcat/Km = 0.5·102 s-1·M-1 at pH = 8.0). The unique feature of AcmB is its capability to convert both testosterone derivatives (C20–C22) as well as steroids substituted at C17 (C27-C30) such as cholest-4-en-3-one or (25R)-spirost-4-en-3-one (diosgenone). Apparent steady-state kinetic parameters were determined for both groups of AcmB substrates. In a batch reactor synthesis, the solubility of water-insoluble steroids was facilitated by the addition of a solubilizer, 2-hydroxypropyl-β-cyclodextrin, and organic co-solvent, 2-methoxyethanol. Catalytic properties characterization of AcmB was tested in fed-batch reactor set-ups, using 0.81 μM of isolated enzyme, PMS and aerobic atmosphere resulting in >99% conversion of the C17–C20 3-ketosteroids within 2 h. Finally, the whole cell E. coli system with recombinant enzyme was demonstrated as an efficient biocatalyst in the synthesis of 1-dehydro-3-ketosteroids.

Anabolic Steroids and Cardiovascular Outcomes: The Controversy.

Anabolic steroids (AS) are synthetic derivatives of the male sex hormone testosterone. The use of AS is not limited to bodybuilders and athletes, but non-athletes also use them. It is used to enhance athletic performance, induce muscle hypertrophy, and augment male sexual characteristics. AS use is associated with a wide range of side effects and potential cardiovascular complications. In this article, we have searched the available literature to investigate the association between AS use and cardiovascular disease (CVD). The results revealed that AS was linked to lipid metabolism derangements, hypertension, coagulation disorders, and cardiomyopathy. We concluded, based on the relevant data, that there was evidence that suggests an association with CVD, primarily myocardial infarction, fatal arrhythmias, and cardiomyopathy in AS users. The general population should be informed of the risk. Also, methods of primary and secondary prevention should be implemented to mitigate the risk of CVD secondary to AS.

The Effects of Sex Hormone Supplementation on Respiratory Function in Male SCI Rats

Sex hormones significantly influence respiratory function and the expression of respiratory neuroplasticity. For example, 17ß‐ estradiol (E2), the most prevalent and neuroactive sex steroid in females, is necessary for expression of phrenic long‐term facilitation (pLTF), a well characterized form of plasticity induced by acute intermittent hypoxia (AIH) in both females and males. To meet this requirement for E2, males aromatize testosterone to E2 in order to express respiratory neuroplasticity. However, on its own (i.e., without aromatase conversion to E2), testosterone has also been shown to enhance breathing. Since both E2 and testosterone have individually demonstrated the capacity to enhance respiratory function, we hypothesized that supplementation of these sex hormones would improve respiratory function in rats 2 weeks post experimental cervical spinal cord injury (SCI). Adult male Sprague‐Dawely rats received a C2 hemi‐section SCI (or Sham laminectomy) and recovered with standard post‐op care. One‐week post‐injury, a controlled‐release hormone pellet was placed subcutaneously in SCI rats to deliver a pre‐determined dose of daily hormone supplementation for the duration of the study. Pellets contained either E2 (β‐estradiol 3‐benzoate, 10 μg/day), 5α‐dihydrotestosterone (DHT), a testosterone derivative that is not converted to E2 (50 μg/day), or blank placebo pellets (50 μg/day). Respiratory function was measured 7 days later (2 weeks post SCI) using Whole‐Body Barometric Plethysmography. Consistent with prior reports, our preliminary findings demonstrated that placebo treated SCI rats breathed with increased respiratory frequency, but reduced tidal volume production. DHT supplementation had little impact on these respiratory parameters, with values remaining similar to placebo control rats. However, one week of E2 supplementation led to reductions in respiratory frequency and increased tidal volume production during baseline room‐air conditions and in response to a strong respiratory challenge (10.5% O2/5% CO2), suggesting improved overall ventilation. These data suggest that estrogen supplementation may facilitate improvements in breathing function in the early stages of spinal cord injury recovery.Support or Funding InformationThis work was supported by funding from The Minnesota Office of Higher Education, The University of Minnesota Division of Physical Therapy, and The University of Minnesota Department of Rehabilitation Medicine

Epistane, an anabolic steroid used for recreational purposes, causes cholestasis with elevated levels of cholic acid conjugates, by upregulating bile acid synthesis (CYP8B1) and cross-talking with nuclear receptors in human hepatocytes.

Anabolic-androgenic steroids are testosterone derivatives, used by body-builders to increase muscle mass. Epistane (EPI) is an orally administered 17α-alkylated testosterone derivative with 2a-3a epithio ring. We identified four individuals who, after EPI consumption, developed long-lasting cholestasis. The bile acid (BA) profile of three patients was characterized, as well the molecular mechanisms involved in this pathology. The serum BA pool was increased from 14 to 61-fold, basically on account of primary conjugated BA (cholic acid (CA) conjugates), whereas secondary BA were very low. In in vitro experiments with cultured human hepatocytes, EPI caused the accumulation of glycoCA in the medium. Moreover, as low as 0.01μM EPI upregulated the expression of key BA synthesis genes (CYP7A1, by 65% and CYP8B1, by 67%) and BA transporters (NTCP, OSTA and BSEP), and downregulated FGF19. EPI increased the uptake/accumulation of a fluorescent BA analogue in hepatocytes by 50-70%. Results also evidenced, that 40μM EPI trans-activated the nuclear receptors LXR and PXR. More importantly, 0.01μM EPI activated AR in hepatocytes, leading to an increase in the expression of CYP8B1. In samples from a human liver bank, we proved that the expression of AR was positively correlated with that of CYP8B1 in men. Taken together, we conclude that EPI could cause cholestasis by inducing BA synthesis and favouring BA accumulation in hepatocytes, at least in part by AR activation. We anticipate that the large phenotypic variability of BA synthesis enzymes and transport genes in man provide a putative explanation for the idiosyncratic nature of EPI-induced cholestasis.

Impact of long-term androgenic anabolic steroid use on a critically-ill patient following traumatic injury

Introduction: Androgenic-anabolic steroids (AAS) are synthetic derivatives of testosterone often used to enhance athletic performance. While short and long-term complications of AAS are recognised clinically, their impact on management of intensive care unit patients after traumatic injury is poorly defined. Furthermore, the implication for glucocorticoid administration is unclear. Objectives/Aims: Here we describe the use of glucocorticoids for haemodynamic support in a long-term AAS user admitted to the intensive care unit (ICU) after traumatic injury. Methods: A 44-year-old male with history significant for depression and AAS use presented to the emergency department after a motor vehicle accident. Injuries included grade 4 liver laceration, rib fractures 5-7, grade 2 right kidney laceration, open dislocation of the right fourth digit, dilation of ascending aorta with aortic valve stenosis, left ventricular hypertrophy, and comminuted complex tear of left ilium, acetabulum and inferior public ramus. He was transferred to the ICU where his course was complicated by hemodynamic instability, acute renal failure requiring renal replacement therapy (RRT), acute respiratory distress syndrome requiring mechanical ventilation and sepsis. Results: During ICU admission, norepinephrine and vasopressin were prescribed per institution guidelines. Hydrocortisone was added for increased vasopressor requirements and concerns of cortisol depletion with previous AAS use and worsening sepsis. Despite transition to midodrine with RRT, norepinephrine was restarted on hospital day 15 for ongoing adrenal insufficiency. Vasopressin and hydrocortisone were also reinitiated for haemodynamic lability, and hydrocortisone tapered over a 14-day course was required. Conclusion: Long-term users of AAS are at risk for complications during hospitalisation, specifically adrenal insufficiency. Use of a prolonged course of glucocorticoid therapy may be necessary for haemodynamic support in patients requiring vasopressor therapy.

Enzymatic Synthesis of Anabolic Steroid Glycosides by Glucosyltransferase from Terribacillus sp. PAMC 23288.

The application of steroids has steadily increased thanks to their therapeutic effects. However, alternatives are required due their severe side effects; thus, studies on the activities of steroid derivatives are underway. Sugar derivatives of nandrolone, which is used to treat breast cancer, as well as cortisone and prednisone, which reduce inflammation, pain, and edema, are unknown. We linked O-glucose to nandrolone and testosterone using UDP-glucosyltransferase (UGT-1) and, then, tested their bioactivities in vitro. Analysis by NMR showed that the derivatives were 17β-nandrolone β-D-glucose and 17β-testosterone β-D-glucose, respectively. The viability was higher and cytotoxicity was evident in PC12 cells incubated with rotenone and, testosterone derivatives, compared to the controls. SH-SY5Y cells incubated with H2O2 and nandrolone derivatives remained viable and cytotoxicity was attenuated. Both derivatives enhanced neuronal protective effects and increased the amounts of cellular ATP.

Profiling of anabolic androgenic steroids and selective androgen receptor modulators for interference with adrenal steroidogenesis

Anabolic-androgenic steroids (AAS) are testosterone derivatives developed for steroid-replacement and treatment of debilitating conditions. They are widely used by athletes in elite sports and bodybuilding due to their muscle-building and performance-enhancing properties. Excessive AAS use is associated with cardiovascular diseases, mood changes, endocrine and metabolic disorders; however, the underlying mechanisms remain unknown. Selective androgen receptor modulators (SARMs) aim to reduce adverse androgenic effects, while maximizing anabolic effects. This study assessed potential steroidogenic disturbances of 19 AAS and 3 SARMs in human adrenocortical carcinoma H295R cells, comparing basal and forskolin-activated states by mass spectrometry-based quantification of nine major adrenal steroids. Mesterolone, mestanolone and methenolone increased mineralocorticoid but decreased adrenal androgen production, indicating CYP17A1 dysfunction. Cell-free activity assays failed to detect direct CYP17A1 inhibition, supported by molecular modeling. The mRNA expression levels of 3β-HSD2, CYP17A1, CYP21A2, CYP11B1 and CYP11B2 were unaffected, suggesting indirect inhibition involving post-translational modification and/or impaired protein stability. Clostebol and oxymetholone decreased corticosteroid but increased dehydroepiandrosterone biosynthesis in H295R cells, suggesting CYP21A2 inhibition, sustained by molecular modeling. These AAS did not affect the expression of key steroidogenic genes. None of the SARMs tested interfered with steroidogenesis. The chosen approach allowed the grouping of AAS according to their steroidogenic-disrupting effects and provided initial mechanistic information. Mesterolone, mestanolone and methenolone potentially promote hypertension and cardiovascular diseases via excessive mineralocorticoid biosynthesis. Clostebol and oxymetholone might cause metabolic disturbances by suppressing corticosteroid production, resulting in adrenal hyperplasia. The non-steroidal SARMs exhibit an improved safety profile and represent a preferred therapeutic option.

Androstadienone resulted in a rightward shift of women's preference for sexually dimorphic male faces across the continuum of femininity-masculinity

As a derivative of testosterone, androstadienone (AND) can influence human psychological and physiological states. To explore the influence of AND on women's preferences for male sexual dimorphic faces in a mate-choice context, we asked 52 females in the luteal phase to choose one from four sexual dimorphic male faces in a long-term and short-term context while inhaling 250 μm of AND or a placebo odor on two consecutive days. Results revealed that participants had a greater and lesser preference for the +30% masculinized and 60% feminized faces, respectively, while inhaling AND, as compared to when inhaling the placebo. The AND intervention resulted in a rightward shift of the women's preference for male sexual dimorphic faces across the continuum of femininity-masculinity. The current findings highlight that AND may influence women's preference for male sexually dimorphic faces in a mate-choice context.

Improved Method for Differentiation of Synthetic and Natural Endogenous Anabolic Steroids using Gas Chromatography Isotope Ratio Mass Spectrometry (GC/C/IRMS) followed by Two-Fold High Performance Liquid Chromatography (HPLC) Cleanup Method: A Perspective

Abstract: Background: Androgenic Anabolic Steroids (AAS) are also synthetic derivatives of testosterone, modified to improve its anabolic actions. The misuse of AAS is of particular concern in sports and society. Gas chromatography-mass spectrometry had some limitations and allows identification and characterization of steroids and their metabolites in the urine but may not be able to distinguish between pharmaceutical (Exogenous) and endogenous origin. Thus, it is of great importance to discriminate endogenous steroids such as testosterone or testosterone prohormones from their chemically identical synthetic copies. The abuse of Androgenic Anabolic Steroids (AAS) by sports person is banned by World Anti-doping Agency (WADA) as per the WADA Prohibited list 2019. Methodology: The gas chromatography-combustion/isotope ratio mass spectrometry (GC/C/IRMS) technique differentiates between natural and synthetic endogenous steroids by comparing compounds specific 13C/12C ratio. However, the analytes have to be efficiently isolated and purified prior to GC/C/IRMS analysis. Results and Discussion: HPLC Cleanup method prior to analysis by GC−C/IRMS needs to be developed and validated for discriminating the origin of anabolic androgenic steroids. These methods involves the solid-phase extraction, enzymatic hydrolysis with β-glucuronidase, HPLC-fractionation for the cleanup and analysis by GC−C/IRMS. The difference (Δ13C) of urinary δ13C values between synthetic analogues and Endogenous Reference Compounds (ERC) by GC-C/IRMS would be used to elucidate the origin of steroids. The present perspective gives an overview of the use of anabolic-androgenic steroids in sport and methods used in anti-doping laboratories for their detection in urine, with special emphasis on GC−C/ IRMS technique after two-fold HPLC cleanup. Key words: Endogenous Steroids, Exogenous Steroids, HPLC Cleanup, Doping, GC/C/ IRMS.