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OBJECTIVE: Human choriogonadotrophin (hCG) treatment of gonadotrophin-deficient infertile men uses hCG of urinary (uhCG) or recombinant (rhCG) origin, but these treatments have not been compared nor are there studies defining rhCG dosing in men. DESIGN: hCG products were studied in randomized cross-over single-dose studies of standard (Study 1, 1500 IU and 62.5 µg, respectively) or high (Study 2, 5000 IU and 250 µg) dose and a multi-dose population pharmacology study of hCG use. PARTICIPANTS: Eight (Study 1) and seven (Study 2) volunteers in cross-over and 52 gonadotrophin-deficient men in the multi-dose study MEASUREMENTS: In cross-over studies, serum testosterone (T), dihydrotestosterone (DHT) and estradiol by liquid chromatography-mass spectrometry (LCMS) and serum hCG, LH, FSH, SHBG and T (observational study) by immunoassays. RESULTS: After standard and high-dose injection, serum hCG and testosterone responses had similar timing and peak concentrations except for a mildly lower early (<48 h) serum testosterone with uhCG. In the multi-dosing study, both hCGs had similar pharmacokinetics (pooled half-life 5.8 days, p < .001), while serum testosterone concentrations were stable after injection and did not differ between hCG products. Bench testing verified that 20% of pens from 4/10 individuals were used inappropriately. CONCLUSIONS: Although hCG pharmacokinetics are not formally bioequivalent, the similar pharmacodynamic effects on serum testosterone indicate that at the doses tested both hCGs provide comparable clinical effects. The starting dose of rhCG for treating gonadotrophin-deficient men should be 62.5 µg (6 clicks) of the rhCG pen.
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Gonadotropina Coriónica , Estudios Cruzados , Proteínas Recombinantes , Testosterona , Humanos , Masculino , Gonadotropina Coriónica/administración & dosificación , Gonadotropina Coriónica/orina , Testosterona/sangre , Testosterona/administración & dosificación , Testosterona/orina , Adulto , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacocinética , Hormona Luteinizante/sangre , Hormona Luteinizante/orina , Dihidrotestosterona/sangre , Dihidrotestosterona/orina , Estradiol/sangre , Relación Dosis-Respuesta a Droga , Hormona Folículo Estimulante/sangre , Hormona Folículo Estimulante/orina , Adulto Joven , Persona de Mediana Edad , Infertilidad Masculina/tratamiento farmacológico , Infertilidad Masculina/orina , Infertilidad Masculina/sangre , Globulina de Unión a Hormona Sexual/análisisRESUMEN
The steroidal module of the athlete biological passport (ABP) targets the use of pseudo-endogenous androgenous anabolic steroids in elite sport by monitoring urinary steroid profiles. Urine and blood samples were collected weekly during two consecutive oral contraceptive pill (OCP) cycles in 15 physically active women to investigate the low urinary steroid concentrations and putative confounding effect of OCP. In urine, testosterone (T) and epitestosterone (E) were below the limit of quantification of 1 ng/ml in 62% of the samples. Biomarkers' variability ranged between 31% and 41%, with a significantly lesser variability for ratios (except for T/E [41%]): 20% for androsterone/etiocholanolone (p < 0.001) and 25% for 5α-androstane-3α,17ß-diol/5ß-androstane-3α,17ß-diol (p < 0.001). In serum, markers' variability (testosterone: 24%, androstenedione: 23%, dihydrotestosterone: 19%, and T/A4: 16%) was significantly lower than in urine (p < 0.001). Urinary A/Etio increased by >18% after the first 2 weeks (p < 0.05) following withdrawal blood loss. In contrast, serum T (0.98 nmol/l during the first week) and T/A4 (0.34 the first week) decreased significantly by more than 25% and 17% (p < 0.05), respectively, in the following weeks. Our results outline steroidal variations during the OCP cycle, highlighting exogenous hormonal preparations as confounder for steroid concentrations in blood. Low steroid levels in urine samples have a clear negative impact on the subsequent interpretation of steroid profile of the ABP. With a greater analytical sensitivity and lesser variability for steroids in healthy active women, serum represents a complementary matrix to urine in the ABP steroidal module.
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Doping en los Deportes , Humanos , Femenino , Esteroides/orina , Testosterona/orina , Dihidrotestosterona/orina , AnticoncepciónRESUMEN
The steroidal module of the athlete biological passport (ABP) introduced by the World Anti-Doping Agency (WADA) in 2014 includes six endogenous androgenic steroids and five of their concentration ratios, monitored in urine samples collected repeatedly from the same athlete, whose values are interpreted by a Bayesian model on the basis of intra-individual variability. The same steroid profile, plus dihydrotestosterone (DHT) and DHEA, was determined in 198 urine samples collected from an amateur marathon runner monitored over three months preceding an international competition. Two to three samples were collected each day and subsequently analyzed by a fully validated gas chromatography-mass spectrometry protocol. The objective of the study was to identify the potential effects of physical activity at different intensity levels on the physiological steroid profile of the athlete. The results were interpreted using principal component analysis and Hotelling's T2 vs Q residuals plots, and were compared with a profile model based on the samples collected after rest. The urine samples collected after activity of moderate or high intensity, in terms of cardiac frequency and/or distance run, proved to modify the basal steroid profile, with particular enhancement of testosterone, epitestosterone, and 5α-androstane-3α,17ß-diol. In contrast, all steroid concentration ratios were apparently not modified by intense exercise. The alteration of steroid profiles seemingly lasted for few hours, as most of the samples collected 6 or more hours after training showed profiles compatible with the "after rest" model. These observations issue a warning about the ABP results obtained immediately post-competition.
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Carrera , Esteroides/orina , Teorema de Bayes , Deshidroepiandrosterona/orina , Dihidrotestosterona/orina , Doping en los Deportes , Ejercicio Físico , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Masculino , Persona de Mediana Edad , Análisis de Componente Principal/métodos , Detección de Abuso de Sustancias/métodosRESUMEN
In this work, male rats were exposed to multiple phthalate esters (MIXPs) in a long-term low-dose model for the early evaluation of reproductive toxicity. An ananlysis method with better sensitivity, accuracy and precision was established to determine the five sex hormones (androstenedione, testosterone, dehydroepiandrosterone, dihydrotestosterone, and estrone) in collected urine samples. The results showed that all the analytes in the MIXPs treated group changed in a time-dependent manner. Specifically, estrone significantly decreased from the 30th day and the other four changed from the 30th day and then significantly increased on the 60th day, while no obvious changes were found in the control group. Therefore, a possible way was provided for the early evaluation of male reproductive toxicity induced by Phthalate esters (PEs) . The reliability of judgment was improved by observing the changes of five target hormones simultaneously. Furthermore, good compliance was predicted for the practical application due to the noninvasive and convenient urine sample collection.
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Androstenodiona/orina , Deshidroepiandrosterona/orina , Dihidrotestosterona/orina , Ésteres/toxicidad , Estrona/orina , Ácidos Ftálicos/toxicidad , Plastificantes/toxicidad , Testosterona/orina , Animales , Masculino , Ratas , Ratas Sprague-Dawley , Reproducción/efectos de los fármacos , Testículo/efectos de los fármacos , Testículo/metabolismoRESUMEN
Background: The impact of testosterone (T) treatment on antidoping detection tests in female-to-male (F2M) transgender men is unknown. We investigated urine and serum sex steroid and luteinizing hormone (LH) profiles in T-treated F2M men to determine whether and, if so, how they differed from hypogonadal and healthy control men. Method: Healthy transgender (n = 23) and hypogonadal (n = 24) men aged 18 to 50 years treated with 1000 mg injectable T undecanoate provided trough urine and blood samples and an additional earlier postinjection sample (n = 21). Healthy control men (n = 20) provided a single blood and urine sample. Steroids were measured by mass spectrometry-based methods in urine and serum, LH by immunoassay, and uridine 5'-diphospho-glucuronosyltransferase 2B17 genotype by polymerase chain reaction. Results: Urine LH, human chorionic gonadotropin, T, epitestosterone (EpiT), androsterone (A), etiocholanolone (Etio), A/Etio ratio, dehydroepiandrosterone (DHEA), dihydrotestosterone (DHT), and 5α,3α- and 5ß,3α-androstanediols did not differ between groups or by time since last T injection. Urine T/EpiT ratio was <4 in all controls and 12/68 (18%) samples from T-treated men, but there was no difference between T-treated groups. Serum estradiol, estrone, and DHEA were higher in transgender men, and serum T and DHT were higher in earlier compared with trough blood samples, but serum LH, follicle-stimulating hormone, and 3α- and 3ß,5α-diols did not differ between groups. Conclusion: Urine antidoping detection tests in T-treated transgender men can be interpreted like those of T-treated hypogonadal men and are unaffected by time since last T dose. Serum steroids are more sensitive to detect exogenous T administration early but not later after the last T dose.
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Andrógenos/metabolismo , Estrógenos/metabolismo , Hipogonadismo/tratamiento farmacológico , Testosterona/análogos & derivados , Transexualidad/tratamiento farmacológico , Adolescente , Adulto , Andrógenos/sangre , Andrógenos/orina , Androsterona/sangre , Androsterona/orina , Deshidroepiandrosterona/sangre , Deshidroepiandrosterona/orina , Dihidrotestosterona/sangre , Dihidrotestosterona/orina , Estradiol/sangre , Estradiol/orina , Estrógenos/sangre , Estrógenos/orina , Estrona/sangre , Estrona/orina , Humanos , Hipogonadismo/sangre , Hipogonadismo/orina , Hormona Luteinizante/sangre , Hormona Luteinizante/orina , Masculino , Espectrometría de Masas , Persona de Mediana Edad , Testosterona/sangre , Testosterona/uso terapéutico , Testosterona/orina , Personas Transgénero , Transexualidad/sangre , Transexualidad/orina , Adulto JovenRESUMEN
INTRODUCTION: There has been considerable interest in the interplay between testosterone and cognition. Dihydrotestosterone (DHT), which has been correlated with cognitive function, is significantly reduced with the use of 5 alpha reductase inhibitors (5ARI) for prostatic enlargement. Our objective was to assess whether the use of 5ARIs was associated with an increased risk of incident dementia. METHODS: We used a matched cohort design and linked administrative data from the province of Ontario, Canada. A total of 99 covariates were measured, and a propensity score was used for matching; 81,162 men who used a 5ARIs were matched to an equal number of men who did not. RESULTS: New initiation of 5ARI medication was associated with an increased risk of dementia during the first (HR 2.18, 95% CI 2.01-2.35) and second (HR 1.52, 95% CI 1.39-1.67) year, however this risk was nonsignificant among the men with the longest exposure to 5ARIs (HR 1.06, 95% CI 0.98-1.14). There was no difference in the results between types of 5ARIs. CONCLUSION: As the strength of the association decreased with increased exposure, the higher risk seen in the initial two years likely represents the presentation and treatment of urinary symptoms which coexist with mild cognitive impairment and eventually progresses to a diagnosis of dementia.
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Inhibidores de 5-alfa-Reductasa/uso terapéutico , Demencia/diagnóstico , Demencia/epidemiología , Inhibidores de 5-alfa-Reductasa/efectos adversos , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Demencia/orina , Dihidrotestosterona/antagonistas & inhibidores , Dihidrotestosterona/orina , Humanos , Masculino , Ontario/epidemiología , Factores de Riesgo , Testosterona/antagonistas & inhibidores , Testosterona/orinaRESUMEN
Non-steroidal drugs that increase endogenous testosterone (T) may be used to exploit ergogenic effects of androgens in power sports. While superactive GnRH analog use is suspected, neither screening nor detection tests are developed. This study aimed to determine if (a) stimulation for 5 days by leuprolide (a superactive GnRH analog) of serum and urine steroids and urine LH is reproducible at a 2 week interval, (b) nandrolone decanoate (ND) co-administration masks responses to leuprolide administration, (c) performance of urine measurement of leuprolide and M1, its major metabolite, as a detection test. Healthy men were randomized into a 4 week parallel group, open label clinical study in which all men had daily sc injections of leuprolide (1mg) for 4 days in the 1st and 3rd weeks with hormone-free 2nd and 4th weeks. In the 3rd week, men were randomized to either ND injections or no extra treatment. Serum steroids were determined by liquid chromatography, tandem mass spectrometry (LC-MS), urine steroids by gas chromatography, mass spectrometry (GC-MS), urine leuprolide and M1 by high resolution LC-MS and urine LH by immunoassay. Leuprolide stimulated striking, reproducible increases in serum and urine LH and steroids (serum T, dihydroT (DHT), 3α diol; urine T, epitestosterone (E) and androsterone (A). ND suppressed basal serum T, E2, 3α diol, and urinary E but did not mask or change the magnitude of responses to leuprolide. Urine leuprolide and M1 measurement had 100% sensitivity and specificity in detecting leuprolide administration up to one day after cessation of injections with the detection window between 1 and 3 days after last dose. Screening using urine steroid and LH measurements, optimally by urinary log10(LHxT), correctly classified 82% of urine samples. It is concluded that leuprolide stimulation of endogenous testosterone is reproducible after a 10-day interval, is not masked by ND and is reliably detected by urine leuprolide or M1 measurement for at least 1 day after administration.
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Leuprolida/administración & dosificación , Hormona Luteinizante/sangre , Sustancias para Mejorar el Rendimiento/administración & dosificación , Testosterona/sangre , Adulto , Dihidrotestosterona/sangre , Dihidrotestosterona/orina , Doping en los Deportes , Estradiol/sangre , Estradiol/orina , Humanos , Leuprolida/farmacocinética , Leuprolida/orina , Hormona Luteinizante/orina , Masculino , Persona de Mediana Edad , Nandrolona/análogos & derivados , Nandrolona/farmacología , Nandrolona Decanoato , Sustancias para Mejorar el Rendimiento/farmacocinética , Sustancias para Mejorar el Rendimiento/orina , Testosterona/orina , Adulto JovenRESUMEN
CONTEXT: There is increasing interest in the analysis of waste water at sewage treatment plants to monitor recreational drug use. This technique is limited for novel psychoactive substances (NPS) due to limited knowledge on their human and bacterial metabolism and stability in waste water. Small studies have reported the detection of NPS using pooled anonymous urine samples, which eliminates some of these potential confounders. OBJECTIVE: To determine patterns of recreational drug, including NPS, use by confirming their presence in analysis of pooled urine from portable street urinals across a wide geographical area in central London over a 6-month period. MATERIALS AND METHODS: Pooled anonymous urine samples were collected from 12 four-bay stand-alone portable urinals distributed once a month across central London for six consecutive months. Samples were analysed using high-performance liquid chromatography coupled to high-resolution accurate mass spectrometry (LC-HRAM-MS); acquired data were processed against target compound databases. RESULTS: With regards to Classical Recreational Drugs, there was consistency of detection of cathine, cocaine, morphine, MDMA over the 6 months, with variability of detection of amphetamine, ketamine and cannabis. With regards to NPS, a total of 13 NPS were detected during the six months. Mephedrone and methylhexaneamine were detected consistently each month. Other commonly detected NPS included methiopropamine (5 months), pipradrol (4 months), cathinone (4 months), 5-(2-aminopropyl)benzofuran (3 months) and 4-methyethcathinone (3 months). Of note, methoxetamine and the synthetic cannabinoid receptor agonists were not detected in any samples. DISCUSSION: Previous studies using the same method detected three and five NPS in a nightclub and pissoir setting, respectively, on a single night. The longer sampling time of 6 months has allowed detection of 13 NPS. Of note, mephedrone showed the least month-to-month variation in detection over the 6-month sampling period. With regards to classical recreational drugs, those detected were consistent with use-data from UK population surveys. The only exception is amphetamine which these surveys have shown a steady decline in use since 1996 but our study showed some variation in the frequency of its detection. However, the sampling period was too short and a longer study is needed to detect the trend in decreasing use. CONCLUSION: This study demonstrates that analysis of anonymous pooled urine samples from stand-alone urinals can be used to detect and monitor trends in the use of classical recreational drugs and NPS in a large city centre over time. This technique has the potential to be a novel key indicator alongside other existing indicators to provide a more robust picture of the use of recreational drugs including NPS.
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Drogas Ilícitas/orina , Psicotrópicos/orina , Detección de Abuso de Sustancias , Azaesteroides/orina , Ciclohexanonas/orina , Ciclohexilaminas/orina , Dihidrotestosterona/análogos & derivados , Dihidrotestosterona/orina , Humanos , Reino UnidoRESUMEN
Here we report a new method for oxosteroid identification utilizing "tandem mass tag hydrazine" (TMTH) carbonyl-reactive derivatisation reagent. TMTH is a reagent with a chargeable tertiary amino group attached through a linker to a carbonyl-reactive hydrazine group. Thirty oxosteroids were analysed after derivatisation with TMTH by electrospray ionization mass spectrometry (ESI-MS) and were found to give high ion-currents compared to underivatised molecules. ESI-tandem mass spectrometry (MS/MS) analysis of the derivatives yielded characteristic fragmentation patterns with specific mass reporter ions derived from the TMT group. A shotgun ESI-MS method incorporating TMTH derivatisation was applied to a urine sample.
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Hidrazinas/química , Cetosteroides/orina , Espectrometría de Masa por Ionización de Electrospray/métodos , Corticoesteroides/orina , Andrógenos/orina , Deshidroepiandrosterona/química , Deshidroepiandrosterona/orina , Dihidrotestosterona/química , Dihidrotestosterona/orina , Humanos , Nandrolona/química , Nandrolona/orina , Progestinas/orina , Espectrometría de Masas en Tándem/métodos , Testosterona/química , Testosterona/orinaRESUMEN
Alterations in the maternal excretion of steroids during pregnancy are not restricted to the production of progesterone and estriol by the fetoplacental unit. Although there is a lack of longitudinal data on urinary androgen concentrations during pregnancy, some studies revealed that modifications in the excretions of androgens might be significant. Recently, several testosterone metabolites excreted as cysteine conjugates have been reported in human urine. We conducted a longitudinal study on androgens conjugated with cysteine and major androgens and estrogens excreted as glucuronides in three pregnant women by mass spectrometric techniques. The urinary concentrations obtained in samples weekly collected during each of the three trimesters and samples collected before pregnancy were compared. Results showed a significant increase in urinary estrogens and norandrosterone and a moderate decrease in the urinary concentrations for most of the androgens. The most significant exception to this behavior was the rise observed for epitestosterone glucuronide when comparing basal levels with the first trimester. Cysteinyl conjugates of testosterone metabolites showed a different behavior. Whereas 4,6-androstanedione remained almost constant through the three trimesters, and Δ(6)-testosterone decreased as the majority of androgens, the excretion profile of 1,4-androstanedione notably increased, reaching a maximum at the third trimester. Alterations in the steroid profile are used in doping control analysis for the screening of endogenous anabolic androgenic steroid misuse. In this study, the main parameters proposed for doping control have been determined for basal samples and samples collected in the first trimester and they have been compared. In spite of the limited number of cases, significant variations have been found in all pregnancies studied. These alterations have to be taken into consideration if anabolic steroids are included into the Athlete Biological Passport. This article is part of a Special Issue entitled 'Pregnancy and Steroids'.
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Andrógenos/orina , Cisteína/orina , Adulto , Androstadienos/orina , Androstenodiona/orina , Deshidroepiandrosterona/orina , Dihidrotestosterona/orina , Doping en los Deportes , Estrógenos/orina , Femenino , Cromatografía de Gases y Espectrometría de Masas/normas , Humanos , Estudios Longitudinales , Embarazo , Estándares de Referencia , Valores de Referencia , Espectrometría de Masas en Tándem/normas , Testosterona/análogos & derivados , Testosterona/orinaRESUMEN
A novel and sensitive extraction procedure using maghemite nanoparticles (γ-Fe2O3) modified with sodium dodecyl sulfate (SDS), as an efficient solid phase, was developed for removal, preconcentration and spectrophotometric determination of trace amounts of oxymetholone (OXM), in the presence of mestanolone (MSL). Combination of nanoparticle adsorption and easily magnetic separation was used for the extraction and desorption of OXM. The preparation of γ-Fe2O3 nanoparticles were obtained by co-precipitation method and their surfaces were modified by SDS. The size and properties of the produced γ-Fe2O3 nanoparticles were determined by X-ray diffraction analysis, FT-IR and scanning electron microscopy measurements. OXM and MSL became adsorbed at pH 3.0. The adsorbed drugs were then desorbed and determined spectrophotometrically using a selective complexation reaction for OXM. The calibration graph was linear in the range 15.0-3300.0 ng mL(-1) of OXM with a correlation coefficient of 0.9948. The detection limit of the method for determination of OXM was 4.0 ng mL(-1). The method was applied for the determination of OXM in human urine samples.
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Andrógenos/orina , Dihidrotestosterona/análogos & derivados , Compuestos Férricos/química , Nanopartículas de Magnetita/química , Oximetolona/orina , Andrógenos/química , Calibración , Dihidrotestosterona/química , Dihidrotestosterona/orina , Humanos , Concentración de Iones de Hidrógeno , Límite de Detección , Microscopía Electrónica de Rastreo , Oximetolona/química , Tamaño de la Partícula , Dispersión del Ángulo Pequeño , Dodecil Sulfato de Sodio/química , Extracción en Fase Sólida/métodos , Espectrofotometría/métodos , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos XRESUMEN
BACKGROUND: 5α-Reductase 2 deficiency (5ARD) is a known cause of 46,XY disorders of sex development (DSD). Traditionally, the diagnosis relies on dihydrotestosterone (DHT) measurement, but the results are often equivocal, potentially leading to misdiagnosis. We reviewed alternative approaches for diagnosis of 5ARD. METHODS: We conducted a retrospective review of the results of urinary steroid profiling (USP) by GC-MS and mutational analysis of SRD5A2 [steroid-5-alpha-reductase, alpha polypeptide 2 (3-oxo-5 alpha-steroid delta 4-dehydrogenase alpha 2)] by PCR and direct DNA sequencing of all 46,XY DSD patients referred to our laboratory with biochemical and/or genetic findings compatible with 5ARD. We also performed a literature review on the laboratory findings of all 5ARD cases reported in the past 10 years. RESULTS: Of 16 patients diagnosed with 5ARD between January 2003 and July 2012, 15 underwent USP, and all showed characteristically low 5α- to 5ß-reduced steroid metabolite ratios. Four patients had DHT measured, but 2 did not reach the diagnostic cutoff. In all 12 patients who underwent genetic analysis, 2 mutations of the SRD5A2 gene were detected to confirm the diagnosis. Twenty-four publications involving 149 patients with 5ARD were published in the review period. Fewer than half of these patients had DHT tested. Nearly 95% of them had the diagnosis confirmed genetically. CONCLUSIONS: 5ARD can be confidently diagnosed by USP at 3 months postnatally and confirmed by mutational analysis of SRD5A2. Interpretation of DHT results may be problematic and is not essential in the diagnosis of 5ARD. We propose new diagnostic algorithms for 46,XY DSD.
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3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/deficiencia , Dihidrotestosterona/orina , Trastorno del Desarrollo Sexual 46,XY/enzimología , Trastorno del Desarrollo Sexual 46,XY/orina , Proteínas de la Membrana/deficiencia , 3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/genética , Adolescente , Adulto , Algoritmos , Niño , Preescolar , Análisis Mutacional de ADN , Trastorno del Desarrollo Sexual 46,XY/genética , Cromatografía de Gases y Espectrometría de Masas , Humanos , Lactante , Masculino , Proteínas de la Membrana/genética , Estudios Retrospectivos , Adulto JovenRESUMEN
Dihydrotestosterone is a more potent androgen than testosterone and plays an important role in endocrine function. We demonstrated that, like testosterone, dihydrotestosterone can be oxidized by human cytochrome P450 (P450) 19A1, the steroid aromatase. The products identified include the 19-hydroxy- and 19-oxo derivatives and the resulting Δ(1,10)-, Δ(5,10)-, and Δ(9,10)-dehydro 19-norsteroid products (loss of 19-methyl group). The overall catalytic efficiency of oxidation was ~10-fold higher than reported for 3α-reduction by 3α-hydroxysteroid dehydrogenase, the major enzyme known to deactivate dihydrotestosterone. These and other studies demonstrate the flexibility of P450 19A1 in removing the 1- and 2-hydrogens from 19-norsteroids, the 2-hydrogen from estrone, and (in this case) the 1-, 5ß-, and 9ß-hydrogens of dihydrotestosterone. Incubation of dihydrotestosterone with human liver microsomes and NADPH yielded the 18- and 19-hydroxy products plus the Δ(1,10)-dehydro 19-nor product identified in the P450 19A1 reaction. The 18- and 19-hydroxylation reactions were attributed to P450 3A4, and 18- and 19-hydroxydihydrotestosterone were identified in human plasma and urine samples. The change in the pucker of the A ring caused by reduction of the Δ(4,5) bond is remarkable in shifting the course of hydroxylation from the 6ß-, 2ß-, 1ß-, and 15ß-methylene carbons (testosterone) to the axial methyl groups (18, 19) in dihydrotestosterone and demonstrates the sensitivity of P450 3A4, even with its large active site, to small changes in substrate structure.
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Aromatasa/química , Aromatasa/metabolismo , Citocromo P-450 CYP3A/química , Citocromo P-450 CYP3A/metabolismo , Dihidrotestosterona , Microsomas Hepáticos/enzimología , Aromatasa/genética , Citocromo P-450 CYP3A/genética , Dihidrotestosterona/sangre , Dihidrotestosterona/química , Dihidrotestosterona/orina , Humanos , Hidroxilación , Masculino , Oxidación-ReducciónRESUMEN
Anabolic-androgenic steroids are frequently misused compounds in sports, and they belong to the controlled substances according to the requirements of the World Anti-Doping Agency. The classical techniques of steroid detection are mass spectrometry coupled to gas or liquid chromatography. Biological methods that base on the ability of substances to bind the steroid receptor are not applied in routine doping control procedures so far, but they appear to be useful for characterization of steroid androgenic potential. In this study we used the yeast androgen receptor reporter system (YAS), which in the past has already successfully been applied to both various androgenic substances and also urine samples. Giving attention to the androgenic potential of steroidal dietary supplements, we exemplified the analysis using both mass spectrometry techniques and the YAS-based assay on the product "Syntrax Tetrabol" which was a confiscated dietary supplement and marketed as a steroid precursor. Identification, structure and the kinetic behavior of its excreted metabolites were analyzed by NMR, GC-MS and LC-MS/MS. The androgenic potential of the parent compound as well as its metabolites in urine was evaluated with the help of the YAS. The application of urine samples with a previous deconjugation and the inclusion of urine density values were carried out and led to increased responses on the YAS. Further, the possibility of a complementary application of structure-based instrumental analysis and biological detection of androgenicity with the help of the YAS seems to be desirable and is discussed.
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Anabolizantes/farmacología , Andrógenos/farmacología , Dihidrotestosterona/metabolismo , Dihidrotestosterona/farmacología , Doping en los Deportes/métodos , Detección de Abuso de Sustancias/métodos , Activación Transcripcional/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Dihidrotestosterona/orina , Cromatografía de Gases y Espectrometría de Masas , Humanos , Hidrólisis , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Masculino , Persona de Mediana Edad , Saccharomyces cerevisiae/efectos de los fármacos , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Exposure to androgens during prenatal development shapes both physiological and behavioral developmental trajectories. Notably, in rhesus macaques, prenatal androgen exposure has been shown to increase rough-and-tumble play, a prominent behavioral feature in males during the juvenile period in primates. While macaques are an Old World, polygamous species with marked sexually dimorphic behavior, New World callitrichine primates (marmosets and tamarins) live in cooperative breeding groups and are considered to be socially monogamous and exhibit minimal sexual dimorphism in social play, which suggests that androgen may affect this species in different ways compared to macaques. In addition, we previously described considerable variation in maternal androgen production during gestation in marmosets. Here we tested the association between this variation and variation in offspring rough-and-tumble play patterns in both males and females. We measured testosterone and androstenedione levels in urine samples collected from pregnant marmoset mothers and then observed their offspring's play behavior as juveniles (5-10 months of age). In contrast to findings in rhesus macaques, hierarchical regression analyses showed that higher gestational testosterone levels, primarily in the second semester, were associated with decreased rough-and-tumble play in juveniles, and this relationship appears to be driven more so by males than females. We found no reliable associations between gestational androstenedione and juvenile play behavior. Our findings provide evidence to suggest that normative variation in levels of maternal androgen during gestation may influence developmental behavioral trajectories in marmosets in a way that contradicts previous findings in Old World primates.
Asunto(s)
Andrógenos/orina , Callithrix , Juego e Implementos de Juego , Preñez , Andrógenos/análisis , Animales , Animales Recién Nacidos , Conducta Animal/fisiología , Callithrix/fisiología , Callithrix/psicología , Callithrix/orina , Dihidrotestosterona/orina , Regulación hacia Abajo , Femenino , Masculino , Madres , Embarazo/orina , Preñez/orina , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Efectos Tardíos de la Exposición Prenatal/psicología , Efectos Tardíos de la Exposición Prenatal/orina , Efectos Tardíos de la Exposición Prenatal/veterinaria , Testosterona/orinaRESUMEN
In doping control, an athlete can only be convicted with the misuse with endogenous steroids like testosterone (T), if abnormal values of steroid metabolites and steroid ratios are observed and if the subsequent analysis with isotope ratios mass spectrometry (IRMS) confirms the presence of exogenously administered androgens. In this work, we compare the results of a novel steroid profiling approach with the performance an in-house developed IRMS method. The developed IRMS has the advantage over other methods to be relatively short in time and with target compounds androsterone, etiocholanolone, 5ß-androstane 3α,17ß-diol and 5α-androstane 3α,17ß-diol. Pregnanediol was used as an endogenous reference compound (ERC). Reference limits for the IRMS values were established and applied as decision limits for the evaluation of excretion urine from administration with oral T, T-gel, dihydrotestosterone (DHT) - gel and dehydroepiandrosterone (DHEA). Results indicated the importance of both androstanediols as important IRMS markers where relative values compared to an ERC (Δδ(13)C) yielded better detection accuracy than absolute δ(13)C-values. The detection times of all administered endogenous steroids were evaluated using the proposed thresholds. The results of traditional steroid profiling and a new approach based upon minor steroid metabolites monitoring introduced in a longitudinal framework were evaluated with IRMS. With traditional steroid profiling methods, 95% of the atypical samples could be confirmed whereas an additional 74% of IRMS confirmed was provided by a new biomarkers strategy. These results prove that the other steroid profiling strategies can improve the efficiency in detection of misuse with endogenous steroids.
Asunto(s)
Androstano-3,17-diol/orina , Androsterona/orina , Doping en los Deportes , Etiocolanolona/orina , Cromatografía de Gases y Espectrometría de Masas/métodos , Detección de Abuso de Sustancias/métodos , Adulto , Isótopos de Carbono , Cromatografía Líquida de Alta Presión , Deshidroepiandrosterona/administración & dosificación , Deshidroepiandrosterona/orina , Dihidrotestosterona/administración & dosificación , Dihidrotestosterona/orina , Femenino , Humanos , Masculino , Pregnanodiol/orina , Estándares de Referencia , Valores de Referencia , EstereoisomerismoRESUMEN
Because of their perceived and reported effects on self-image, muscle development, performance, and similar factors, anabolic-androgenic steroids (AAS) and their precursors are among the most abused substances by professional, amateur, and recreational athletes. However, AAS abuse is not limited to athletes, but is also prevalent in the workplace, especially those professions in which image, strength, and endurance are coveted attributes. The detection of many steroids in biological specimens is analogous to the detection of an abused drug such as cocaine. Identification of the parent drug or its characteristic metabolite(s) in a donor's sample with a drug screening technique and confirmation of the drug/metabolite with a suitable alternative technology provides evidence of use. These analyses and subsequent interpretive scenarios become far more complex when the ingested AAS is an endogenous compound such as dehydroepiandrosterone (DHEA), androstenedione (Adione), or dihydrotestosterone (DHT). These compounds and their metabolites are present in specimens such as urine as a course of our natural endocrine function. Therefore, it becomes much more challenging for the laboratory to establish testing and interpretative paradigms that can distinguish "normal" urinary profiles of these steroids and their metabolites from profiles indicative of exogenous use. Distinguishing "normal" from "abnormal" urine profiles is particularly challenging during screening when literally tens of steroids and their metabolites may be tested simultaneously in a single chromatographic analysis. The purpose of this paper is to review the relevant literature about DHEA, Adione, and DHT administration, detection, and interpretation specifically as it relates to changes in the urinary AAS profile that may be identified during the routine laboratory screening of donor urine specimens.
Asunto(s)
Androstenodiona/orina , Deshidroepiandrosterona/orina , Dihidrotestosterona/orina , Doping en los Deportes/prevención & control , Sustancias para Mejorar el Rendimiento/orina , Detección de Abuso de Sustancias/métodos , Biomarcadores/orina , Humanos , Límite de Detección , Estándares de ReferenciaRESUMEN
The natural occurrence of endogenous anabolic steroids together with their availability in different administration forms makes the detection of their misuse a great challenge for doping control laboratories. Nowadays, the detection of endogenous steroids abuse is performed by the analysis of the steroid profile. Recently, androst-1,4-dien-3,17-dione (1,4-AD), androst-4,6-dien-3,17-dione (4,6-AD), 17ß-hydroxy-androst-4,6-dien-3-one (6-T), and androst-15-en-3,17-dione (15-AD) have been described as testosterone (T) metabolites released after basic treatment of the urine. In the present work, the usefulness of these metabolites has been evaluated detecting the use of three different forms of endogenous steroids in a single dose: dihydrotestosterone gel (DHT), oral dehydroepiandrosterone (DHEA), and T gel. After the independent administration of these endogenous steroids, a rise in the value of several of the ratios calculated between the tested metabolites was noticed. For DHT, a small increase was observed for the ratios 1,4-AD/15-AD, 6-T/15-AD and 4,6-AD/15-AD although only for one volunteer. Better results were obtained for oral DHEA and T gel where an increase was observed in all volunteers for several of the tested ratios. The detection time in which the misuse can be detected (DT) has been evaluated using two different approaches: (1) comparison with population based reference limits, and (2) comparison with individual threshold levels. The obtained DTs were compared with the results of previously published markers for the misuse of such substances. When using basic released metabolites, shorter DTs were obtained for DHT, similar DTs for DHEA, and the detectability was substantially improved for T gel.
Asunto(s)
Deshidroepiandrosterona/metabolismo , Deshidroepiandrosterona/orina , Dihidrotestosterona/metabolismo , Dihidrotestosterona/orina , Detección de Abuso de Sustancias/métodos , Testosterona/metabolismo , Testosterona/orina , Administración Oral , Administración Tópica , Adulto , Deshidroepiandrosterona/administración & dosificación , Dihidrotestosterona/administración & dosificación , Doping en los Deportes , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Masculino , Sensibilidad y Especificidad , Testosterona/administración & dosificación , Adulto JovenRESUMEN
New analogues of androgens that had never been available as approved drugs are marketed as "dietary supplement" recently. They are mainly advertised to promote muscle mass and are considered by the governmental authorities in various countries, as well as by the World Anti-doping Agency for sport, as being pharmacologically and/or chemically related to anabolic steroids. In the present study, we report the detection of a steroid in a product seized by the State Bureau of Criminal Investigation Schleswig-Holstein, Germany. The product "1-Androsterone" of the brand name "Advanced Muscle Science" was labeled to contain 100mg of "1-Androstene-3b-ol,17-one" per capsule. The product was analyzed underivatized and as bis-TMS derivative by GC-MS. The steroid was identified by comparison with chemically synthesized 3ß-hydroxy-5α-androst-1-en-17-one, prepared by reduction of 5α-androst-1-ene-3,17-dione with LS-Selectride (Lithium tris-isoamylborohydride), and by nuclear magnetic resonance. Semi-quantitation revealed an amount of 3ß-hydroxy-5α-androst-1-en-17-one in the capsules as labeled. Following oral administration to a male volunteer, the main urinary metabolites were monitored. 1-Testosterone (17ß-hydroxy-5α-androst-1-en-3-one), 1-androstenedione (5α-androst-1-ene-3,17-dione), 3α-hydroxy-5α-androst-1-en-17-one, 5α-androst-1-ene-3α,17ß-diol, and 5α-androst-1-ene-3ß,17ß-diol were detected besides the parent compound and two more metabolites (up to now not finally identified but most likely C-18 and C-19 hydroxylated 5α-androst-1-ene-3,17-diones). Additionally, common steroids of the urinary steroid profile were altered after the administration of "1-Androsterone". Especially the ratios of androsterone/etiocholanolone and 5α-/5ß-androstane-3α,17ß-diol and the concentration of 5α-dihydrotestosterone were influenced. 3α-Hydroxy-5α-androst-1-en-17-one appears to be suitable for the long-term detection of the steroid (ab-)use, as this characteristic metabolite was detectable in screening up to nine days after a single administration of one capsule.
Asunto(s)
Anabolizantes/análisis , Androsterona/análogos & derivados , Suplementos Dietéticos/análisis , Detección de Abuso de Sustancias/métodos , Testosterona/análogos & derivados , Anciano , Anabolizantes/farmacocinética , Androstano-3,17-diol/orina , Androsterona/química , Androsterona/farmacocinética , Androsterona/orina , Dihidrotestosterona/orina , Etiocolanolona/orina , Humanos , Masculino , Testosterona/química , Testosterona/orinaRESUMEN
Testosterone regulates a wide variety of behavioral and physiological traits in male vertebrates. It influences reproductive and aggressive behaviors and is used as a marker of gonadal activity. While testosterone is the primary biologically active male gonadal steroid in the blood, it is metabolized into a variety of related steroids when excreted via urine and feces. To monitor endocrinological profiles studies on wild-living animals primarily rely on non-invasively collected samples such as urine or feces. Since a number of androgen metabolites that are found in high concentrations in these matrices do not stem exclusively from gonadal production, but are also produced by the adrenal cortex, the metabolism and excretion pattern of testosterone and its characteristic metabolites have to be investigated. Here, we compare the levels of 11 androgens and their metabolites in serum and urine (after hydrolytic/solvolytic cleavage of conjugates) from female, and intact and castrated male chimpanzees to investigate whether they were of testicular or adrenal origin. For serum, significant differences in concentrations were found only for native testosterone. For urine, testosterone concentrations showed the largest differences between intact and castrated males, and intact males and females, while no differences were seen between females and castrated males. Epitestosterone levels revealed the same pattern. These differences in urinary concentrations could also be seen for 5α-androstane-3α,17ß-diol (androstanediol), and less clearly for 5α-dihydrotestosterone (5α-DHT), etiocholanolone, and androsterone. In urine of males, significant correlations were found between the levels of testosterone and 5α-androstane-3α,17ß-diol, as well as between testosterone and epitestosterone. Therefore, the clearest urinary markers of gonadal activity in male chimpanzees seems to be testosterone itself.