Context: Testosterone abuse is conventionally assessed by the urinary testosterone/epitestosterone (T/E) ratio, levels above 4.0 being considered suspicious. The large variation in testosterone ...glucuronide (TG) excretion and its strong association with a deletion polymorphism in the uridine diphospho-glucuronosyl transferase (UGT) 2B17 gene challenge the accuracy of the T/E ratio test.
Objective: Our objective was to investigate whether genotype-based cutoff values will improve the sensitivity and specificity of the test.
Design: This was an open three-armed comparative study.
Participants: A total of 55 healthy male volunteers with either two, one, or no allele insertion/insertion, insertion/deletion, or deletion/deletion (del/del) of the UGT2B17 gene was included in the study.
Intervention: A single im dose of 500 mg testosterone enanthate was administered.
Main Outcome Measures: Urinary excretion of TG after dose and the T/E ratio during 15 d were calculated.
Results: The degree and rate of increase in the TG excretion rate were highly dependent on the UGT2B17 genotype with a 20-fold higher average maximum increase in the insertion/insertion group compared with the del/del group. Of the del/del subjects, 40% never reached the T/E ratio of 4.0 on any of the 15 d after the dose. When differentiated cutoff levels for the del/del (1.0) and the other genotypes (6.0) were applied, the sensitivity increased substantially for the del/del group, and false positives in the other genotypes were eliminated.
Conclusions: Consideration of the genetic variation in disposition of androgens will improve the sensitivity and specificity of the testosterone doping test. This is of interest not only for combating androgen doping in sports, but also for detecting and preventing androgen abuse in society.
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.
Abstract
Context
Little is known about how exogenous testosterone (T) affects the steroid profile in women. More knowledge would give the antidoping community keys as to how to interpret tests and ...detect doping.
Objective
This work aimed to investigate the steroid profile in serum and urine in young healthy women after T administration.
Methods
In a randomized, double-blind, placebo-controlled study, 48 healthy young women were assigned to daily treatment with T cream (10 mg) or placebo (1:1) for 10 weeks. Urine and blood were collected before and at the end of treatment. Serum steroids were analyzed with liquid chromatography–tandem mass spectrometry, and urine levels of T, epitestosterone (E), and metabolites included in the Athlete Biological Passport (ABP) were analyzed with gas chromatography–tandem mass spectrometry.
Results
In serum, T and dihydrotestosterone levels increased, whereas sex hormone–binding globulin and 17-hydroxyprogesterone decreased after T treatment as compared to placebo. In urine, T and 5α-androstanediol increased in the T group. The median T increase in serum was 5.0-fold (range, 1.2-18.2) and correlated to a 2.2-fold (range, 0.4-14.4) median increase in T/E in urine (rs = 0.76). Only 2 of the 24 women receiving T reached the T/E cutoff ratio of 4, whereas when the results were added to the ABP, 6 of 15 participants showed atypically high T/E (40%). In comparison, 22/24 women in the T group increased serum T more than 99.9% of the upper confidence interval of nontreated values.
Conclusion
It seems that the T/E ratio is not sufficient to detect exogenous T in women. Serum total T concentrations could serve as a complementary marker of doping.
To evaluate the impact of genetic polymorphisms of UGT enzymes (UGT1A4, UGT2B7, UGT2B15 and UGT 2B17) and the transporter protein ABCB1 on Lamotrigine (LTG) metabolism.
Single nucleotide ...polymorphisms UGT1A4*2 (P24T, c.70C>A), UGT1A4*3 (L48V c.142T>G), UGT2B7*2 (H802Y, c.802C>T), UGT2B15*2 (Y85D, c.253G>T), UGT2B17 deletion and transporters ABC 1236C> T and 3435C> T were determined in 337 Caucasian patients with epilepsy treated with LTG in Denmark. The prospectively collected data included LTG dosage, LTG plasma concentration, 2-N-GLU concentration, sex, smoking habits, concomitant medicine, oral contraceptives (OC).
The non-smokers with LTG monotherapy and LTG polytherapy with other non-interacting drugs NIAEDs (n = 199) were analyzed separately in univariant analyses. LTG ratios (LTG plasma concentration/ (LTG dose/weight)) in patients carrying wild type UGT1A4*2 C-allele were 22% lower than in heterozygous C-carriers (p = 0.013). Patients with UGT2B7*2 polymorphism TT genotype had 1.2-fold higher LTG ratios (p = 0.0078) and 0.78-fold lower GLU/LTG ratio (p = 0.0275) than patients homozygous for the C allele. The similar significant findings were also seen comparing homozygotes (TT) with heterozygotes patients (CT). Individuals homozygous for the UGT2B15*2 T allele displayed 18% lower LTG ratio concentrations than individuals homozygous for the G allele (p = 0.014),while significant difference in GLU/LTG ratio was only seen comparing wild type with homozygous patients (GG versus TT, p = 0.031). A copy number variation gene deletion polymorphism of UGT2B17 showed that individuals devoid of the gene (del/del) exhibited 1.3-fold higher LTG ratio (p = 0.015). For ABCB1c.1236 C>T and ABC1B1c.3435 C>T no associations with LTG and GLU ratios were found.
Sex specific differences in enzyme activity (most prominent effect in women) on LTG metabolism were found for UGT2B15, UGT2B17, UGT1A4 and UGT2B7 polymorphisms.
Multiple regression analysis confirmed the significant effect of OC, VPA and UGT1A4 * 2 and UGT2B7 * 2 on LTG metabolism.
Our study confirms the previous findings that genetic variations in UGT2B7 and UGT1A4 genes are associated with serum LTG concentrations.
Furthermore, our results indicate that it is possible that different UGT genotypes may exert larger impact on LTG metabolism in women than in men.
•Lower enzyme activity was seen on LTG metabolism in patients with UGT1A*2 and UGT2B7*2 polymorphisms.•Sex specific differences in enzyme activitywere found for UGT2B15, UGT2B17, UGT1A4 and UGT2B7 polymorphisms•Multiple regression analysis confirmed the significant effect of OC, VPA and UGT1A4*2 and UGT2B7*2 on LTG metabolism.
The role of endogenous androgens for body composition and physical performance in women athletes is still not elucidated.
To examine the serum androgen profile in relation to body composition and ...physical performance in women Olympic athletes and to compare endocrine variables and body composition to controls.
Cross-sectional study, conducted between 2011 and 2015 at the Women's Health Research Unit, Karolinska University Hospital, Stockholm.
Swedish women Olympic athletes (n=106) and age-matched and body mass index-matched sedentary controls (n=117) were included in the study. Blood sampling was performed in a rested, fasting state for the measurement of serum androgens and their metabolites by liquid chromatography-tandem mass spectrometry. Body composition was determined by dual-energy X-ray absorptiometry (controls n=100, athletes n=65). The athletes performed standardised performance tests (n=59) (squat jump (SJ) and countermovement jump (CMJ).
The athletes demonstrated significantly higher levels of the precursor androgens dehydroepiandrosterone (DHEA) and 5-androstene-3β, 17β-diol (5-DIOL) and the metabolite etiocholanolone glucuronide (Etio-G), significantly lower levels of estrone (p<0.05, respectively), higher bone mineral density (p<0.001) and more lean mass (p<0.001) compared with controls. Serum levels of DHEA, 5-DIOL and Etio-G correlated positively to lean mass variables and physical performance in the athletes. DHEA and lean mass legs explained 66% of the variance in SJ, whereas lean mass explained 52% of the variance in CMJ.
The present data suggest that endogenous androgens are associated with a more anabolic body composition and enhanced performance in women athletes. These results are of importance for the current discussion regarding hyperandrogenism in women athletes.
The large variation in disposition known for most drugs is also true for anabolic androgenic steroids. Genetic factors are probably the single most important cause of this variation. Further, there ...are reasons to believe that there is a corresponding variation in efficacy of doping agents. Doped individuals employ a large variety of doping strategies in respect of choice of substance, dose, dose interval, duration of treatment and use of other drugs for enforcement of effects or correction of side effects.
Metabolic steps up‐stream and down‐stream of testosterone are genetically variable and contribute substantially to the variation in disposition of testosterone, the most common doping agent in sports and in society. Large inter‐ and intra‐ethnic variation in testosterone glucuronidation and excretion is described as well as the pit‐falls in evaluation of testosterone doping test results. The hydrolysis and bioactivation of testosterone enanthate is also genetically variable yielding a 2–3 fold variation in excretion rate and serum concentration, thereby implicating a substantial variation in ‘efficacy’ of testosterone.
Given this situation it is logical to adopt the new findings in the doping control programme. The population based cut‐off level for the testosterone : epitestosterone ratio should be replaced by a Bayesian interpretation of consecutive tests in the same individual. When combined with the above genetic information the sensitivity of the test is considerably improved. The combination of the three approaches should reduce the rate of falsely negative or positive results and the number of expensive follow‐up tests, stipulated by the World Anti‐Doping Agency.
Background:
The second to fourth digit ratio (2D:4D ratio) is suggested to be a negative correlate of prenatal testosterone. Little is known about the role of the 2D:4D ratio in relation to serum and ...urinary androgens for physical performance in female athletes. We aimed to compare the 2D:4D ratio in female Olympic athletes with sedentary controls, and to investigate the 2D:4D ratio in relation to serum and urinary androgens and physical performance in the athletes.
Methods:
This cross-sectional study included 104 Swedish female Olympic athletes participating in power, endurance and technical sports and 117 sedentary controls. The 2D:4D ratio was calculated using direct digit measurements. Serum androgens and urinary androgen metabolites were analyzed by liquid chromatography-tandem mass spectrometry. The athletes performed standardized physical performance tests and body composition was established by dual-energy X-ray absorptiometry.
Results:
The 2D:4D ratio was significantly lower in the athletes compared with controls although serum testosterone levels were comparable between groups and within normal reference values. The 2D:4D ratio correlated negatively with urinary levels of testosterone glucuronide and 5α- and 5βAdiol-17G, whereas there were no correlations to serum androgen levels. Furthermore, the 2D:4D ratio correlated negatively with strength tests and positively with 3,000-meter running in the athletes.
Conclusion:
Female Olympic athletes had a lower 2D:4D ratio, possibly reflecting a higher prenatal androgen exposure, than sedentary controls. Furthermore, the 2D:4D ratio was related to urinary levels of androgen metabolites and physical performance in the athletes but not to serum androgen levels. It is suggested that the 2D:4D ratio could reflect androgen metabolism and may be of importance for sporting success in female athletes.
Anabolic androgenic steroids (AAS) are used for their aesthetic and performance-enhancing effects and are associated with physical and psychological side effects. Behavioural changes/side effects as ...mood swings, aggressiveness, depression, potency problems, anxiety, and emotional coldness have been reported by next of kin to people using AAS.
This phenomenological study is based on the reflective lifeworld research approach. Interviews were conducted with twelve next of kin about their experiences of living close to persons using AAS.
Next of kin to persons using AAS are particularly vulnerable because they experience little opportunity to influence their situation. Their given and safe context is lost, and their lives are circumscribed by feelings of insecurity, fear, powerlessness, and grief. Feelings of loneliness develop when their problems are not noticed by others and support is lacking from family and society.
Our research adds important knowledge on how the use of AAS affects next of kin. Understanding is required to approach the lifeworld of next of kin with flexibility and empathy in their difficulties and vulnerability. Healthcare professionals and other concerned professions need to be aware of next of kin existential needs to be able to meet and support them in their life situation.
The World Anti‐Doping Agency (WADA) has recently implemented dried blood spots (DBSs) as a matrix for doping control. However, specifications regarding the analysis of the class of prohibited ...substances called erythropoietin (EPO) receptor agonists (ERAs) from DBSs are not yet described. The aim of this study was to find optimal conditions (sample volume and storage) to sensitively detect endogenous erythropoietin (hEPO) and prohibited ERAs from DBSs and compare detection limits to WADA‐stipulated minimum required performance levels (MRPLs) for ERAs in serum/plasma samples. Venous whole blood was spotted onto Whatman 903 DBS cards with primarily 60 μl of blood, but various volumes from 20 to75 μl were tested. All samples were immunopurified with MAIIA EPO Purification Gel kit (EPGK) and analysed with sodium N‐lauroylsarcosinate polyacrylamide gel electrophoresis (SAR‐PAGE) and Western blot. Sixty‐microliter DBSs allowed the detection of the four main ERAs (BRP, NESP, CERA and EPO‐Fc) at concentrations close to WADA's MRPLs described for 500 μl of serum/plasma. Different storage temperatures, from −20°C to 37°C, were evaluated and did not affect ERA detection. A comparison of the detection of endogenous EPO from the different anti‐doping matrices (urine, serum and DBSs produced from upper arm capillary blood) from five participants for 6 weeks was performed. Endogenous EPO extracted from DBSs showed intra‐individual variations in male and female subjects, but less than in urine. Doping controls would benefit from the stability of ERAs on DBSs: It can be a complementary matrix for ERA analysis, particularly in the absence of EPO signals in urine.
The main findings from this project about erythropoietin (EPO) receptor agonists (ERAs) in dried blood spots (DBSs) are as follows:
ERA concentrations close to the World Anti‐Doping Agency's minimum required performance levels could be detected from 60‐μl DBSs.
ERAs are confirmed to be stable on DBSs.
Endogenous erythropoietin can be detected from an upper arm volumetric device with a polymer support.
Dyslipidemia in metabolic syndrome may introduce an underestimation of the risk for cardiovascular disease (CVD) using Low-Density Lipoprotein-Cholesterol (LDL-C) as a surrogate marker. Recently, ...non-High-Density Lipoprotein-Cholesterol (non-HDL-C), Apolipoprotein B (ApoB) and remnant-Cholesterol (remnant-C) have been suggested as better biomarkers for dyslipidemia. In addition, the microbial metabolites trimethylamine-N-oxide (TMAO), betaine and choline have been associated with CVD and suggested as markers for dysbiosis. There is a lack of knowledge on potential alterations in these biomarkers during the menstrual cycle. The aim of this single center, prospective non-interventional study, was to investigate variations in biomarkers of dyslipidemia and dysbiosis in healthy volunteers during the menstrual cycle.
Serum samples were collected from 17 healthy, regularly menstruating women during two menstrual cycles, including the follicular, ovulatory and luteal phases. Levels of lipoproteins, lipoprotein ratios and microbial metabolites were analyzed in a total of 90 samples (30 complete menstrual cycles).
ApoB, ApoB/HDL and non-HDL-C/HDL ratios were significantly higher in the follicular phase compared to the ovulatory and luteal phases (p < 0.05). Remnant-C were higher during the luteal phase (p < 0.05). TMAO did not vary during the different phases and did not correlate with estrogen levels.
Our data support that biomarkers for dyslipidemia vary during the menstrual cycle. Thus, to avoid an underestimation of cardiovascular risk, sampling during the follicular phase, when levels of pro-atherogenic lipids are higher, may be considered.
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