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Problem: With menopause, the risk of cardiovascular diseases (CVD) increases significantly. A possible molecular mechanism is an estrogen-related change in fat metabolism. Endurance training has been demonstrated to reduce the risk of CVD and to have an impact on fat metabolism (FM). This study aims to analyze the ability of pre- (preW) and postmenopausal women (postW) of comparable body mass index (BMI) to activate their FM during endurance training.Methods: 12 preW and 12 postW were included. Serum Triglyceride, LDL, HbA1C, estradiol and body composition data were determined. The respiratory quotient (RQ) was determined during moderate 30-minute exercise (60% of the 4 mmol threshold) on an ergometer.Results: While the BMI of preW and postW was comparable, body fat (BF) (p=0.001), lean body mass (LBM) (p=0.001) and abdominal girth (AG) (p=0.003) were significantly different. Significant group effects could also be identified in HbA1c (p=0.001), cholesterol (p=0.001) and LDL (p=0.000) serum concentrations. RQ decreased during 30 minutes of cycling in preW and increased in postW (p=0.010) over time.Discussion: The higher AG and BF and the lower LBM demonstrates the change in body composition in postW. An accumulation of fat, especially in the trunk region, goes along with an increase of CVD in postW, even with a normal BMI.Conclusion: It is evident that postW show altered lipid metabolism compared to preW.Key Words: Menopause, Cardiovascular Risk Factor, Aerobic Training, Metabolism

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ABSTRACT Dietary analysis using δ15N values of human remains such as bone and hair is usually based on general principles and limited data sets. Even for modern humans, the direct ascertainment of dietary δ15N is difficult and laborious, due to the complexity of metabolism and nitrogen fractionation, differing dietary habits and variation of δ15N values of food items. The objective of this study was to summarize contemporary regional experimental and global literature data to ascertain mean representative δ15N values for distinct food categories. A comprehensive data set of more than 12,000 analyzed food samples was summarized from the literature. Data originated from studies dealing with (1) authenticity tracing or origin control of food items, and (2) effects of fertilization or nutrition on δ15N values of plants or animals. Regional German food δ15N values revealed no major differences compared with the mean global values derived from the literature. We found that, in contrast to other food categories, historical faunal remains of pig and poultry are significantly enriched in 15N compared to modern samples. This difference may be due to modern industrialized breeding practices. In some food categories variations in agricultural and feeding regimens cause significant differences in δ15N values that may lead to misinterpretations when only limited information is available. Am J Phys Anthropol 152:58-66, 2013. © 2013 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

The side effects and safety of cannabidiol (CBD) products are currently discussed in different contexts. Of all adverse effects, hepatotoxic effects have been reported most frequently in previous studies. However, the threshold for liver toxicity of CBD in humans is uncertain due to the lack of adequately designed studies in humans below the lowest observed adverse effect level (LOAEL) of 300 mg/day. In a randomised, three-arm, double-blind, crossover study, the effects of two CBD products (oil and solubilisate (solu) containing 60 mg CBD) were investigated during a high-intensity exercise protocol. Seventeen well-trained subjects (26±4 years, 181±5 cm, 85.6±9.4 kg) participated in the intervention. All subjects were healthy and had no physiological or psychological injuries. Participants were divided into advanced (Ad) and highly advanced (Hi) athletes … They consumed 60 mg of the compound in each microcycle over 7 days. To evaluate possible effects of short-term repeated use of 60 mg CBD on oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), gamma-glutamyl transferase (GGT) and creatinine (CREA) were analysed before and after each microcycle. GOT increased significantly in both performance levels of the placebo groups (Ad: p≤0.001; Hi: p=0.003). This increase was significantly reduced in the Ad group by both CBD oil (p=0.050, ES=0.66) and CBD solu (p=0.027; ES=0.75). GPT also increased significantly in both placebo groups (Ad: p≤0.001; Hi: p=0.032). This increase was significantly reduced in the Ad group by both CBD oil (p=0.027; ES=0.75) and CBD solu (p=0.023; ES=0.77). These effects were not observed in the Hi group for either parameter. Our results show that short-term repeated use of 60 mg CBD can inhibit exercise-induced liver activity. Furthermore, under the conditions of the present study, there was no evidence for hepatotoxic effects of oral intake of CBD at 60 mg for seven days. Nevertheless, despite the inhibitory effect on exercise-induced liver activity, the study provides evidence for the pharmacological effects of CBD on the liver even at low CBD dose and does not exclude adverse effects in sensitive individuals., Competing Interests: Declarations. Conflict of interest: The authors declare no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Rapid regeneration after intense exercise is essential for competitive athletes. Based on this assumption, supplementation strategies, focusing on food supplements, are increasing to improve the recovery processes. One such supplement is cannabidiol (CBD) which is gaining more attention in competitive sports. However, the evidence is still lacking and there are no data available about the effect of a short-term chronic application., Methods: A three-arm double-blind cross-over study was conducted to determine the effects of two different CBD products on performance, muscle damage and inflammatory processes in well-trained athletes. In total 17 subjects took successfully part in this study. Each subject underwent the six-day, high-intensity training protocol three times. After each training session, each subject took either a placebo or a CBD product (60 mg of oil or solubilisate). Between the intervention phases, at least four weeks of washout period was conducted. Before and after the training protocols the performance capacity in countermovement jump (CMJ), back squat (BS), bench press (BP) and 1-mile run were measured and biomarkers for muscle damage (creatine kinase, myoglobin), inflammatory processes (interleukin 6 and 10) and immune cell activity (ratios of neutrophil granulocytes, lymphocytes and, platelets) were analyzed. For statistical analyses, the current version of R and a linear mixed model was used., Results: It could identify different effects of the training protocol depending on performance level (advanced or highly advanced athletes) ( p  < .05). Regardless of the performance level, muscle damage and a reduction in performance could be induced by the training protocol. Only CBD oil was associated with a reduction in myoglobin concentration ( p  < .05) in advanced athletes. Concerning immune activity, a significant decrease in platelets lymphocyte ratios was observed in advanced athletes after placebo treatment ( p  < .05). CBD oil application showed a slight inhibitory effect ( p  < .10). Moreover, the reduction in performance differs between the performance levels. A significant decrease in CMJ was observed in advanced athletes and a decreasing trend in BS was observed in highly advanced athletes after placebo treatment ( p  < 0.10). Both CBD products do not affect performance parameters. For inflammatory parameters, no effects were observed., Conclusion: It was found that the performance level of the subjects was a decisive factor and that they responded differently to the training protocol and the CBD application. However, no clear effects of either CBD product were found and further research is needed to identify the long-term effects of CBD application.

Objective: The influence of the menstrual cycle on practical power performance such as barbell back squats and jumping performance in women has not yet been fully investigated. In addition, the performance level of athletes has not been considered in previous studies. This study aimed to investigate the influence of different cycle phases on acute back squat performance, jumping ability and psychological state concerning the performance level., Methods: 24 female strength athletes (age: 25.2 ± 3.3 years; height: 169.5 ± 3.4 cm; body weight: 67.7 ± 7.3 kg) were recruited for the study. Level of performance was classified according to Santos et al. (intermittent (n = 13), advanced (n = 6), highly advanced (n = 5)). Participants were tested for 1RM barbell back squat and jumping performance (countermovement and squat jump) as well as two questionnaires assessing their psychological states in the menses (M), late follicular phase (FP) and mid-luteal phase (LP) in three MC. Saliva estradiol and progesterone concentrations with a menstrual cycle diary were used to confirm a normal MC. A principal components analysis for power performance, well-being, relaxation and alertness was carried out and a linear mixed model was used for statistical evaluation., Results: No significant differences were found between the MC phases in performance scores (p > 0.05), readiness (p > 0.05) and alertness (p > 0.05). However, a high correlation between MC phases, performance level and back squat performance was detected. Correlation analyses indicate that a higher performance level results in a higher variation depending on the MC of the squat performance. For well-being, a significantly lower score was detected in M than in FP and LP., Conclusion: In general the performance score of the lower body is not influenced by the MC. If strength performance and jumping ability are considered separately, there are indications that strength capability is influenced at a higher performance level. In addition, individual variance was also observed, so this should also be considered. However, further studies are needed to confirm this assumption due to the small sample sizes of the individual performance levels., Trial Registration: German registry for clinical studies (DRKS00034816, Date: 08/01/2024)., (© 2024. The Author(s).)

Background: The consumption of dietary supplements (DS) is widespread among the general population and competitive athletes. However, only a few competitive athletes seek information from experts about the effects and use of DS. Furthermore, it is currently unknown whether certain sports have a higher affinity for DS than others., Methods: This study aimed to identify differences between different sports categories and subgroups that may have a very high affinity for DS. For this purpose, competitive athletes were surveyed. The survey included the type of sport, the training frequency, the number of competitions, the consumption behaviour of five DS categories (general health, regeneration promotion, performance enhancement, booster, and weight loss) as well as personal data such as biological sex and age. Subsequently, correlations, configural frequencies (CFA), and multiple correspondence analyses (MCA) were used to identify subgroups with a high affinity of consumption behaviour., Results: A total of 409 questionnaires could be evaluated. It was found that all DS categories except weight loss were related. In addition, it was observed that in sports from the power category and from the endurance category, there was even higher consumption behaviour than in other sports categories. Male power athletes in particular have a higher affinity for consuming DS than other subgroups., Conclusions: This study shows that there is a clear different consumption behaviour depending on the type of sport. Male power athletes in particular are the subgroup with the greatest consumption behaviour and therefore require special education on the effects and use of DS. This subgroup in particular should receive increased attention in counselling on DS to minimise the possible risks of DS use.

Background: Resistance training (RT) is effective in counteracting the age- and menopause-related loss of muscle mass (MM) and strength in middle-aged women (40-60 years). Research on RT with free weights is limited in pre- and post-menopausal women. Based on this, a 20-week training intervention was conducted with this population to investigate the effects of systematic RT with free weights on strength capacity and body composition., Method: Forty-one healthy women (52.0 ± 3.6 years) participated in this study. After 10-week control phase (no RT, T0-T1) followed by a 10-week intervention phase (T1-T2) with RT twice a week and 6-8 sets of each muscle per week. Subjects were randomly assigned to a low-intensity (50% 1-RM) or moderate-intensity (75% 1-RM) RT group and divided into pre-menopausal and post-menopausal according to their hormone profile. Fat-free mass (FFM), MM, fat mass (FM), muscle thickness (Vastus lateralis (VL), Rectus femoris (RF), Triceps brachii (TB)), grip strength, 1-RM squat and bench press were assessed before and after each phase. Statistical analysis was performed using a linear mixed model to account for fixed (time and group) and random (individual) effects., Results: A total of 31 women successfully completed the study. No injuries occurred during the intervention. Significant increases in 1-RM squat and bench press were observed in all groups. No interaction effect was observed for the strength parameters. In pre-menopausal women, FFM, MM and RF muscle thickness increased significantly, while VL showed a trend. These effects were not present in post-menopausal women regardless of RT intensity., Conclusion: RT with free weight is safe and effective for middle-aged women to increase 1-RM. Hypertrophy effects were found exclusively in pre-menopausal women. To achieve hypertrophy and/or body composition changes in post-menopausal women, larger training volumes (> 6-8 sets/muscle per week) are likely required., (© 2023. BioMed Central Ltd., part of Springer Nature.)

The use of plant steroids to improve physical health and performance is becoming increasingly popular. One of these plant steroids is diosgenin, which is mainly available in fenugreek. As a result, some studies have been conducted to improve physical health. Fenugreek extracts are also becoming increasingly popular in the context of athletic performance. Based on these assumptions, a systematic review with meta-analysis was conducted to evaluate the promoting effects of fenugreek on strength performance, body composition, and hormone concentration. Four databases were screened according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The current version of ReviewManager (RevMan) was used for the statistical evaluation. Seven studies with 449 participants (378 male, 71 female) met the inclusion criteria. A small effect of fenugreek was detected for total testosterone (standard mean difference (SMD): 0.32; 95% confidence interval (CI): 0.09 0.55), free testosterone (SMD: 0.24; 95% CI: -0.04, 0.52), lean body mass (SMD: 0.19; 95% CI: -0.10, 0.49), fat mass (SMD: -0.19; 95% CI: -0.44, 0.05), and leg press performance (SMD: 0.22; 95% CI: -0.02, 0.47), in male athletes. The meta-analysis shows that chronic application of fenugreek has performance-enhancing and anabolic effects in male athletes, but no statements can be made for female athletes., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

The age-related loss of muscle mass promotes many impairments. Training and protein supplementation are suggested to prevent muscle wasting, but recommendations for all populations are not based on scientific evidence. This study combines protein/carbohydrate supplementation (PCS) and training for seniors and postmenopausal women. Project A: 51 postmenopausal women (PMW, 57.3 ± 3.0 years old) underwent health-oriented training (12 weeks, moderate-strength training + moderate-endurance training). The intervention group (IG) additionally received 110 g sour milk cheese (SMC) and toast. Project B: 25 women and 6 men (65.9 ± 4.9 years old) performed intense sling training (12 weeks). The IG additionally received 110 g SMC, toast, and buttermilk. Strength was tested before and after in both studies. Project A: there was significant increase in strength, no additional effect of PCS, and a reduction in body fat in the controls. Project B: there was significant increase in strength, significant additional effects of PCS for trunk strength, and a significant reduction in body weight. Combining training and PCS may counteract strength loss. Combined endurance/resistance training is recommended to PMW for whom the benefits of PCS are restricted. Aged subjects may benefit from PCS when training intensely, but these benefits may be strongly individual.

Background: Chronic obstructive pulmonary disease (COPD) is one of the world's most common diseases and reasons for death. Systemic consequences, especially reduced muscle strength, muscle mass and muscle function, are common and contribute to worsening prognosis and increasing morbidity and mortality. There is strong evidence that physical activity and strength training are effective in prolonging life and lead to better quality of life. Numerous studies have shown that ingestion of protein and carbohydrates after strength training can increase regeneration of strength in young athletes. Recently, we demonstrated that the same effect can be achieved with these macronutrients administered in a meal. Until now, it is not clear if patients with COPD, integrated in regular physical training, respond similarly., Methods: Prescribed strength training, consisting of two sets circular training with machines for big muscle groups was supplemented with a meal rich in protein and carbohydrates. Changes in maximum strength after 24 h were investigated to find out about the impact of this meal on physical capacity. A pilot study was conducted with pragmatic cross-over design., Results: With nutritive intervention, strength in both knee extensor and chest press were significantly higher than in control training., Conclusion: The study showed beneficial effects for the intake of protein and carbohydrates in changes in maximum strength. For now, the underlying mechanism remains unclear. Clinical relevance needs further research. The study design and study protocol can be used for further studies with only small adaptions.

Background: Hormones like testosterone play a crucial role in performance enhancement and muscle growth. Therefore, various attempts to increase testosterone release and testosterone concentration have been made, especially in the context of resistance training. Among practitioners, sexual activity (coitus and masturbation) a few hours before training is often discussed to result in increases of testosterone concentration and thus promote muscle growth. However, there is no evidence to support this assumption and the kinetics of the testosterone and cortisol response after sexual activity have not been adequately investigated. Therefore, the aim of this pilot-study was to examine the kinetics of hormone concentrations of total testosterone, free testosterone and cortisol and their ratios after masturbation. In a three-arm single blinded cross-over study, the effects of masturbation with visual stimulus were compared to a visual stimulus without masturbation and the natural kinetics in healthy young men., Results: The results showed a significant between-condition difference in free testosterone concentrations. Masturbation (p < 0.01) and a visual stimulus (p < 0.05) may seem to counteract the circadian drop of free testosterone concentrations over the day. However, no statistical change was observed in the ratios between total testosterone, free testosterone and cortisol., Conclusions: It can be assumed that masturbation may have a potential effect on free testosterone concentrations but not on hormonal ratios. However, additional studies with larger sample sizes are needed to validate these findings., (© 2021. The Author(s).)

Cannabidiol supplements (CBD) are increasingly consumed by athletes to improve regeneration. However, the evidence for the pro-regenerative effects of CBD in sports is quite limited. Therefore, our aim was to investigate the effects of a single CBD supplementation in a six-arm placebo-controlled crossover study after resistance training on performance and muscle damage. Before and after the resistance training, one-repetition maximum in the back squat (1RM BS), countermovement jump (CMJ), and blood serum concentrations of creatine kinase (CK) and myoglobin (Myo) were measured in healthy, well-trained participants. 16 out of 21 participants completed the study and were included in the analysis. In 1RM BS, a significant decrease was observed after 24 h ( p < 0.01) but not after 48 and 72 h. A significant group difference was detected after 72 h ( p < 0.05; ES = 0.371). In CMJ, no significant changes were observed. The CK and Myo concentrations increased significantly after 24 h (CK: p < 0.001; Myo: p < 0.01), 48 h (CK: p < 0.001; Myo: p < 0.01) and 72 h (CK: p < 0.001; Myo: p < 0.001). After 72 h, significant group differences were observed for both muscle damage biomarkers (CK: p < 0.05 ES = 0.24; Myo: p < 0.05; ES = 0.21). The results show small but significant effects on muscle damage and recovery of squat performance after 72 h. However, more data are required for clearer statements concerning potential pro-regenerative effects of CBD supplementation after resistance training.

Isotope ratio mass spectrometry (IRMS) has been established in doping control analysis to identify the endogenous or exogenous origin of a variety of steroidal analytes including the 19-norsteroid metabolite norandrosterone (NorA). NorA can be found naturally in human urine in trace amounts due to endogenous demethylation or in situ microbial degradation. The administration of nortestosterone (nandrolone) or different prohormones results in the excretion of urinary NorA. Usually, this can be detected by IRMS due to differing δ 13 C values of synthetic 19-norsteroids compared to endogenous reference compounds. The consumption of uncastrated pig edible parts like offal or even meat may also lead to a urinary excretion of NorA. In order to determine the δ 13 C values of such a scenario, urine samples collected after consumption of a wild-boar-testicle meal were analyzed. IRMS revealed highly enriched δ 13 C values for urinary NorA, which could be related to the completely corn-based nutrition of the animal. Isotopic analysis of the boar's bristles demonstrated a dietary change from C 3 -based forage, probably in winter and spring, to a C 4 -based diet in the last weeks to months prior to death. These results supported the interpretation of an atypical test result of a Central European athlete's doping control sample with δ 13 C values for NorA of -18 ‰, most probably caused by the consumption of a wild boar ragout. As stated before, athletes should be fully aware of the risk that consumption of wild boar may result in atypical or even adverse analytical findings in sports drug testing., (© 2018 John Wiley & Sons, Ltd.)

Natural stable isotope ratios (δ 15 N) of humans can be used for nutritional analyses and dietary reconstruction of modern and historic individuals and populations. Information about an individual's metabolic state can be obtained by comparison of tissue and dietary δ 15 N. Different methods have been used to estimate dietary δ 15 N in the past; however, the validity of such predictions has not been compared to experimental values. For a total of 56 meals and 21 samples of 24-h diets, predicted and experimental δ 15 N values were compared. The δ 15 N values were predicted from self-recorded food intake and compared with experimental δ 15 N values. Predicted and experimental δ 15 N values were in good agreement for meals and preparations (r = 0.89, p < .001) as well as for the 24-h diets (r = 0.76, p < .001). Dietary δ 15 N was mainly determined by the amount of fish, whereas the contribution of meat to dietary δ 15 N values was less pronounced. Prediction of human dietary δ 15 N values using standardised food records and representative δ 15 N data sets yields reliable data for dietary δ 15 N intake. A differentiated analysis of the primary protein sources is necessary when relating the proportion of animal-derived protein in the diet by δ 15 N analysis.

Rationale: Natural stable nitrogen isotope ratios (δ 15 N) are frequently used for the determination of provenance and dietary assessment of recent and ancient humans. Although individual δ 15 N values typically correspond to the dietary δ 15 N composition, they are also affected by metabolic conditions. Preferred matrices for the measurement of human δ 15 N values have been hair, nail or blood. The goal of this study was to validate a novel approach for the assessment of the δ 15 N values from urinary urea, the principal end-product of human N metabolism., Methods: The method, which involves the precipitation of urea from urine using xanthydrol, was validated using fortified urea solutions. Intra- and inter-individual variance of the δ 15 N values of urinary urea was determined from samples obtained from multiple human subjects., Results: Precipitation with xanthydrol did not alter the δ 15 N values of urea. The mean δ 15 N value in urinary urea from human subjects from Germany was +4.4 ± 0.6 ‰, which corresponds to the estimated dietary composition. It falls below previously reported δ 15 N values for human tissue and blood samples. Longitudinal analyses over 7 days illustrate short-time changes linked to varying protein intake., Conclusions: Our results indicate that δ 15 N values can be measured reliably from human urine and that the method is suitable to monitor rapid dietary and metabolic changes of an individual. Our findings further confirm that urinary urea is depleted in 15 N compared with human tissue but within the range of the δ 15 N composition of the diet. Copyright © 2017 John Wiley & Sons, Ltd., (Copyright © 2017 John Wiley & Sons, Ltd.)

Rationale: Natural stable carbon (δ(13)C) and nitrogen isotope ratios (δ(15)N) of humans are related to individual dietary habits and environmental and physiological factors. In forensic science the stable isotope ratios of human remains such as hair and nail are used for geographical allocation. Thus, knowledge of the global spatial distribution of human δ(13)C and δ(15)N values is an essential component in the interpretation of stable isotope analytical results., Methods: No substantial global datasets of human stable isotope ratios are currently available, although the amount of available (published) data has increased within recent years. We have herein summarised the published data on human global δ(13)C andδ(15)N values (around 3600 samples) and added experimental values of more than 400 additional worldwide human hair and nail samples. In order to summarise isotope ratios for hair and nail samples correction factors were determined., Results: The current available dataset of human stable isotope ratios is biased towards Europe and North America with only limited data for countries in Africa, Central and South America and Southeast Asia. The global spatial distribution of carbon isotopes is related to latitude and supports the fact that human δ(13)C values are dominated by the amount of C4 plants in the diet, either due to direct ingestion as plant food, or by its use as animal feed. In contrast, the global spatial distribution of human δ(15)N values is apparently not exclusively related to the amount of fish or meat ingested, but also to environmental factors that influence agricultural production., Conclusions: There are still a large proportion of countries, especially in Africa, where there are no available data for human carbon and nitrogen isotope ratios. Although the interpretation of modern human carbon isotope ratios at the global scale is quite possible, and correlates with the latitude, the potential influences of extrinsic and/or intrinsic factors on human nitrogen isotope ratios have to be taken into consideration., (Copyright © 2015 John Wiley & Sons, Ltd.)

The leptin resistant Zucker diabetic fatty (ZDF) rats are hyperphagic and become obese, but whereas the males develop type 2 diabetes mellitus (T2DM), the females remain euglycaemic. As estrogen deficiency is known to increase the risk of developing T2DM, we evaluated the role of ER subtypes alpha and beta in the development of glucose tolerance in leptin resistant ovariectomized (OVX) ZDF rats. At least six rats per group were treated with either vehicle (OVX), 17β-estradiol (E2), ER subtype-selective agonists (Alpha and Beta), or genistein (Gen) for 17 weeks. At the end of the treatment period a glucose tolerance assay was performed and the metabolic flux of (13)C-glucose for the E2 group was investigated. OVX ZDF rats treated with E2, Alpha, Beta, and Gen tolerated the glucose significantly better than untreated controls. E2 treatment increased absorbance/flux of (13)C-glucose to metabolic relevant tissues such liver, adipose tissue, gastrocnemius, and soleus muscle. Moreover, whereas Alpha treatment markedly increased mRNA expression of GLUT4 in gastrocnemius muscle, Beta treatment resulted in the largest fiber sizes of the soleus muscle. Treatment with Gen increased both the mRNA expression of GLUT 4 and the fiber sizes in the skeletal muscle. In addition, E2 and Alpha treatment decreased food intake and body weight gain. In summary, estrogen-improved glucose absorption is mediated via different molecular mechanisms: while activation of ER alpha seems to stimulate muscular GLUT4 functionality, activation of ER beta results in a hypertrophy of muscle fibers. In addition, selective activation of ER alpha decreased food intake and body weight gain. Our data further indicate that ER subtype-selective agonists and genistein improve systemic glucose tolerance also in the absence of a functional leptin signaling pathway., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The present study aimed to investigate the relationship between the mechanical load during resistance exercise and the elicited physiological responses. Ten resistance-trained healthy male subjects performed 1 set of resistance exercise each at 55%, 70%, and 85% of 1 repetition maximum for as many repetitions as possible and in 4 training modes: 4-1-4-1 (4 s concentric, 1 s isometric, 4 s eccentric, and 1 s isometric successive actions), 2-1-2-1, 1-1-1-1, and explosive (maximum velocity concentric). Mean concentric power and total concentric work were determined. Oxygen uptake (V̇O2) was measured during exercise and for 30 min post exercise. Total volume of consumed oxygen (O2 consumed) and excess post-exercise oxygen consumption (EPOC) were calculated. Maximum blood lactate concentration (LAmax) was also determined. V̇O2 exhibited a linear dependency on mean concentric power. Mean concentric power did not have a detectable effect on EPOC and LAmax. An augmentation of total concentric work resulted in significant linear increase of O2 consumed and EPOC. Total concentric work caused a significant increase in LAmax. In general, a higher mechanical load induced a larger physiological response. An increase in mean concentric power elicited higher aerobic energy turnover rates. However, a higher extent of total concentric work augments total energy cost covered by oxidative and (or) glycolytic pathways.

The molecular mechanisms underlying the effects of selective ER subtype activation on lipogenesis, adipogenesis, lipid utilization and storage as well as glucose metabolism are currently largely unknown and were analyzed in female OVX Wistar rats on a high-fat diet. Rats received estradiol (E2), ER subtype-selective agonists (Alpha and Beta), and genistein (Gen) for 10 weeks. In adipose tissue, treatment with E2, Alpha, and Beta significantly decreased lipogenic (SREBP-1c, FAS) and adipogenic genes (LPL, PPAR gamma). In liver and skeletal muscle of E2-, Alpha-, Beta-, and Gen-treated animals, lipogenesis and triglyceride accumulation were significantly reduced. Increased hepatic and muscular PPAR gamma mRNA expression was observed in untreated, Beta- and Gen-treated animals, which correlates with increased hepatic glucose uptake. However, only untreated animals showed impaired insulin sensitivity compared to all other groups. Therefore, PPAR gamma up-regulation in untreated animals suggests a compensatory mechanism, probably due to increased triglyceride accumulation in non-adipose tissues. Beta- and Gen-treated animals may benefit from the anabolic potency of ER beta that ameliorates lipid and glucose utilization in muscle. Activation of either ER subtype reduces fat enrichment and improves insulin sensitivity. Depending on the investigated tissue, different molecular pathways seem to be involved., (Copyright © 2013. Published by Elsevier Ireland Ltd.)

The administration of musk extract, that is, ingredients obtained by extraction of the liquid secreted from the preputial gland or resulting grains of the male musk deer (eg, Moschus moschiferus), has been recommended in Traditional Chinese Medicine (TCM) applications and was listed in the Japanese pharmacopoeia for various indications requiring cardiovascular stimulation, anti-inflammatory medication or androgenic hormone therapy. Numerous steroidal components including cholesterol, 5α-androstane-3,17-dione, 5β-androstane-3,17-dione, androsterone, etiocholanolone, epiandrosterone, 3β-hydroxy-androst-5-en-17-one, androst-4-ene-3,17-dione and the corresponding urea adduct 3α-ureido-androst-4-en-17-one were characterised as natural ingredients of musk over several decades, implicating an issue concerning doping controls if used for the treatment of elite athletes. In the present study, the impact of musk extract administration on sports drug testing results of five females competing in an international sporting event is reported. In the course of routine doping controls, adverse analytical findings concerning the athletes' steroid profile, corroborated by isotope-ratio mass spectrometry (IRMS) data, were obtained. The athletes' medical advisors admitted the prescription of TCM-based musk pod preparations and provided musk pod samples for comparison purposes to clarify the antidoping rule violation. Steroid profiles, IRMS results, literature data and a musk sample obtained from a living musk deer of a local zoo conclusively demonstrated the use of musk pod extracts in all cases which, however, represented a doping offence as prohibited anabolic-androgenic steroids were administered.

Δ(13)C and δ(13)C values of endogenous urinary steroids represent physiological random variables. Measurement uncertainty and biological scatter likewise contribute to the variances. The statistical distributions of negative controls are well investigated, but there is little knowledge about the corresponding distributions of steroid-users. For these reasons valid discrimination of steroid users from non-users by (13)C/(12)C analysis of endogenous steroids requires elaborate statistical treatment. Corresponding Bayesian approaches are presented following an introduction to the rationale. The use of mixture models appears appropriate. The distribution of routine data has been deconvolved and characterized accordingly. The mixture components, which presumably represent steroid users and non-users, exhibit considerable overlap. The validity of a given result depends on both the analytical uncertainty and the prior probability of doping offenses. Low analytical uncertainties but high prior probabilities facilitate valid detection of doping offenses. Two recommendations can be deduced. First, before starting an (13)C/(12)C analysis, any initial suspicion should be well-substantiated. This precludes use of permissive criteria derived from the steroid profile. Secondly, knowledge of relevant (13)C/(12)C distributions is required. This must cover representative numbers of authentic steroid users. Finally, it is desirable that the conditional probability for steroid administration rather than the measurement uncertainty is calculated and reported. This quantity possesses superior validity and it is largely independent of laboratory bias. The findings suggest and facilitate flexible handling of decision limits. Proposals for the evaluation of stable isotope data are presented., (Copyright © 2012 John Wiley & Sons, Ltd.)

The history of isotope ratio mass spectrometry (IRMS) is briefly described. It is shown that the fundamental design of isotope ratio mass spectrometers has not changed since the 1940s. The basic findings concerning the natural variation of isotope abundances even date back to the 1930s. Recent improvements in the methodology mainly concern online coupling and analytical peripherals. The nature of isotopic scales necessitates a specific terminology which is unfamiliar to many analysts. However, corresponding guidelines exist that should be adopted by the anti-doping community. Currently, steroids represent the only group of compounds routinely analyzed by IRMS in doping-control. Suggestions are made in respect to a harmonized terminology concerning the nature and origins of steroids., (Copyright © 2012 John Wiley & Sons, Ltd.)

Technical advances are being made in many areas of biotechnology and genetics that are facilitating the detection of doping in sport. These improvements have been catalyzed by the need to counter the ever-increasing sophistication of the community of athletes and their retinues who are intent on the illicit use of physical, pharmacological and genetic tools and methods to enhance athletic performance, in contravention of established international ethical and legal standards and of international treaty. The methods described in this article present a partial and general picture of only some of these advances.

The determination of salbutamol and its glucuronide in human urine following the inhalative and oral administration of therapeutic doses of salbutamol preparations was performed by means of direct urine injection utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and employing d(3)-salbutamol and d(3)-salbutamol glucuronide as internal standards. Unconjugated salbutamol was detected in all administration study urine samples. Salbutamol concentrations following inhalation were commonly (99%) below 1000 ng/ml whereas values after oral administration frequently (48%) exceeded this threshold. While salbutamol glucuronide was not detected in urine samples collected after inhalation of the drug, 26 out of 82 specimens obtained after oral application contained salbutamol glucuronide with a peak value of 63 ng/ml. The percentage of salbutamol glucuronide compared to unconjugated salbutamol was less than 3%. Authentic doping control urine samples indicating screening results for salbutamol less than 1000 ng/ml, showed salbutamol glucuronide concentrations between 2 and 6 ng/ml, whereas adverse analytical findings resulting from salbutamol levels higher than 1000 ng/ml, had salbutamol glucuronide values between 8 and 15 ng/ml. The approach enabled the rapid determination of salbutamol and its glucuronic acid conjugate in human urine and represents an alternative to existing procedures since time-consuming hydrolysis or derivatization steps were omitted. Moreover, the excretion of salbutamol glucuronide in human urine following the administration of salbutamol was proven., (Copyright © 2011 John Wiley & Sons, Ltd.)

Boldenone (androsta-1,4-dien-17beta-ol-3-one, Bo) is an anabolic steroid known to have been used in cattle breeding or equine sport as a doping agent for many years. Although not clinically approved for human application, Bo or its main metabolite 5beta-androst-1-en-17beta-ol-3-one (BM1) were detected in several doping control samples. For more than 15 years the possibility of endogenous Bo production in human beings has been discussed. This is a challenging issue for doping control laboratories as Bo belongs to the list of prohibited substances of the World Anti-Doping Agency and therefore the chance for false positive testing is significant. By GC/C/IRMS (gas chromatography/combustion/isotope ratio mass spectrometry) it should be possible to analyze the (13)C/(12)C ratio of either Bo or BM1 and to distinguish whether their source is endogenous or exogenous. Therefore a method was developed to determine the (13)C/(12)C ratios of Bo, BM1, pregnanediol, androsterone, etiocholanolone, and testosterone from a single urine specimen. The validity of the method was ensured by repeated processing of urine fortified with 2-50 ng/mL Bo and BM1. The specificity of the method was ensured by gas chromatography/mass spectrometry determinations. Out of 23 samples investigated throughout the last four years, 11 showed (13)C/(12)C ratios of Bo or BM1 inconsistent with an exogenous origin. Two of these samples were collected from the same athlete within a one-month interval, strongly indicating the chance of endogenous Bo production by this athlete.

Testosterone is the principal male sex hormone. As with all natural steroids, it is biosynthesized from cholesterol. Phase I metabolism employs some very specific enzymes and pathways. Phase II metabolism and excretion follow more general patterns. The effects of testosterone are twofold: anabolic and androgenic. Because of its anabolic effects, testosterone is frequently abused in sports. Because of its endogenous nature, testosterone doping is difficult to detect. The standard procedure is based on the evaluation of the urinary steroid profile. Conspicuous samples then are submitted to compound-specific (13)C/(12)C analysis. Synthetic and endogenous steroids differ in this measure. Numerous xenobiotic compounds have been derived from testosterone. The modifications typically aim at a reduction of the androgenic properties while maintaining the anabolic potential. Most of these compounds have been withdrawn from the legal market. However, they are found to be illicitly added to otherwise inefficient nutritional supplements. These products represent a major problem to doping control. Recently, clinical trials with selective androgen receptor modulators have been started., (Copyright 2010 Elsevier Inc. All rights reserved.)

Adrenosterone (androst-4-ene-3,11,17-trione, 11-oxoandrostenedione) is an endogenous steroid hormone that has been promoted as a dietary supplement capable of reducing body fat and increasing muscle mass. It is proposed that adrenosterone may function as an inhibitor of the 11beta-hydroxysteroid dehydrogenase type 1 enzyme (11beta-HSD1), which is primarily responsible for reactivation of cortisol from cortisone. The urinary metabolism of adrenosterone was investigated, after a single oral administration in two male subjects, by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Substantially increased excretion of 11beta-hydroxyandrosterone, 11beta-hydroxyetiocholanolone, 11-oxoandrosterone and 11-oxoetiocholanolone was observed. Minor metabolites such as 3alpha,17beta-dihydroxy-5beta-androstan-11-one, 3alpha-hydroxyandrost-4-ene-11,17-dione and 3alpha,11beta-dihydroxyandrost-4-en-17-one were also identified. The exogenous origin of the most abundant adrenosterone metabolites was confirmed by GC-C-IRMS according to World Anti-Doping Agency criteria. Through analysis of a reference population data set obtained from urine samples provided by elite athlete volunteers (n = 85), GC-MS doping control screening criteria are proposed: 11beta-hydroxyandrosterone concentration greater than 10 000 ng/mL (specific gravity adjusted to 1.020) or 11beta-hydroxyandrosterone/11beta-hydroxyetiocholanolone ratio greater than 20.Urine samples fulfilling these screening criteria may be subjected to GC-C-IRMS analysis for confirmation of adrenosterone administration., (Copyright (c) 2010 John Wiley & Sons, Ltd.)

Epitestosterone (17alpha-hydroxy-androst-4-en-3-one, EpiT) belongs to the list of prohibited substances of the World Anti-Doping Agency (WADA). Although it possesses no anabolic effect, it is presumed to be misused by athletes in order to mask administration of testosterone (T) by lowering the urinary T/EpiT ratio.To improve detection, an excretion study with 40 mg of orally administered EpiT was conducted focusing on the metabolites of EpiT: 5alpha- and 5beta-androstane-3alpha,17 alpha-diol (5aEpiD and 5bEpiD). A reference population of n = 74 volunteers was investigated to elucidate the urinary concentrations of these steroids.In order to prove whether an unusual finding in urinary concentrations or ratios is due to an illicit intake of steroids or due to physiological elevation, determination of carbon isotope ratios is advisable. A method for isotope ratio determination was developed to enable (13)C/(12)C ratios of EpiT, 5bEpiD, 5aEpiD, pregnanediol and androsterone and etiocholanolone to be measured from a single urine specimen. The method's validity was tested by applying linear mixing models and specificity was ensured by means of gas chromatography/mass spectrometry analysis. delta(13)C values at natural levels were determined with a reference population and both Delta values and corresponding reference limits were calculated.Considering the implemented EpiT-metabolites, a more than twofold extension of the detection time of EpiT administration was achieved with both the urinary concentration thresholds and the (13)C/(12)C ratios., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

The abundances of the stable isotopes of the bioelements are not constant. Subtle, but significant, variations may be induced by physical, physiological and biochemical processes. These variations may be detected and quantified. Often, isotope fingerprints are characteristic of certain processes and may reveal information concerning the sources and origins of compounds of interest. Moreover, natural variabilities of stable isotopes may be exploited in order to perform tracer experiments. The most accurate technology to perform stable isotope analysis is (gas) isotope ratio MS (IRMS). Compound-specific approaches employ hyphenation of GC and LC to IRMS. In these approaches, complete conversion to simple gases prior to MS is required. Analysis by stable isotope ratio spectroscopy currently approaches the accuracy of IRMS. However, for bioanalytical projects, it is still predominantly confined to material synthetically enriched with stable isotopes.

The carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) stable isotope ratios of human hair can be used for the interpretation of dietary habits and nutritional status in contemporary or past populations. Although the results of bulk or segmental isotope ratio analysis of human hair have been used for the reconstruction of an individual's diet for years, only limited data of controlled dietary changes on the carbon and nitrogen isotopic composition of human hair are available. Hair of four individuals, two males and two females, who participated in a dietary change experiment for 28 days was segmentally analysed for delta(13)C and delta(15)N. The dietary change included a change from C3 to C4 plant enriched diets and a simultaneous replacement of terrestrial animal products by marine products. This resulted in an increase in delta(13)C(diet) of +8.5 to +9.9 per thousand and in delta(15)N(diet) of +1.5 to +2.2 per thousand. All subjects showed significant increases in delta(13)C(hair) and delta(15)N(hair) during the dietary change period, although no subject reached a new steady state for either carbon or nitrogen. The change in delta(15)N(hair) was faster than the change in delta(13)C(hair) for all individuals. The magnitude of change of the isotopic composition during the dietary change period could be attributed to the degree of physical activity of the individuals, with a higher physical activity resulting in a faster change., (2009 John Wiley & Sons, Ltd.)

The development and application of a combined gas chromatography/thermal conversion/isotope ratio mass spectrometry (GC/TC/IRMS) method for D/H ratio determination of endogenous urinary steroids are presented. The key element in sample preparation was the consecutive cleanup with high-performance liquid chromatography of initially native and subsequently acetylated steroids. This strategy enabled sufficient cleanup off all target analytes for determination of their respective D/H values. Ten steroids (11beta-hydroxyandrosterone, 5alpha-androst-16-en-3alpha-ol, pregnanediol, androsterone, etiocholanolone, testosterone, epitestosterone, 5alpha-androstan-3alpha,17beta-diol, 5beta-androstan-3alpha,17beta-diol and dehydroepiandrosterone) were measured from a single urine specimen. Depending on the biological background, the determination limit for all steroids ranged from 10 to 15 ng/mL for a 20 mL specimen. The method was validated by application of linear mixing models on each steroid and covered repeatability and reproducibility. The specificity of the procedure was ensured by gas chromatography/mass spectrometry (GC/MS) analysis of the sample using equivalent chromatographic conditions to those employed in the GC/TC/IRMS measurement. Within the sample preparation, no isotopic fractionation was observed, and no amount-dependent shift of the D/H ratios during the measurement was noticed. Possible memory effects occurring during IRMS measurements were corrected by applying a simple rule of proportion. In order to determine the naturally occurring D/H ratios of all implemented steroids, a population of 18 male subjects was analyzed. Relevant mean Delta values among selected steroids were calculated which allowed us to study the metabolic pathways and production sites of all the implemented steroids with additional consideration of the corresponding (13)C/(12)C ratios., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

The urinary phase II metabolites of norsteroids, 19-norandrosterone, 19-noretiocholanolone and 19-norepiandrosterone glucuronide and sulphate, were analyzed in samples collected during the pregnancy, following the administration of norsteroids or the consumption of edible parts of non-castrated pig and in athletes' samples in which they were found during routine controls. The level of the sulfo- and glucuroconjugated metabolites was precisely determined by GC/HRMS, after selective hydrolysis. The goal was to evaluate whether the fine analysis of the norsteroid conjugates produced and excreted in different conditions would show a pattern that could be linked to their origin. The delta (13)C values of the metabolites formed following the ingestion of edible parts of non-castrated pig were measured by isotope ratio mass spectrometry. Our results indicated that it is not possible to determine the origin of the urinary metabolites based upon the sole evaluation of the different metabolites and conjugates. The GC/C/IRMS is the only method permitting to distinguish between the exogenous and endogenous origin of the metabolites.

In order to detect the misuse of endogenous anabolic steroids such as testosterone by athletes a total of n = 1734 suspicious urine samples were investigated by gas chromatography/combustion/isotope ratio mass spectrometry throughout the years 2005, 2006 and 2007. The (13)C/(12)C ratio of a target substance (androsterone, a testosterone metabolite) was compared to the (13)C/(12)C ratio of an endogenous reference compound (11beta-hydroxyandrosterone).N = 1340 samples were investigated due to elevated testosterone/epitestosterone ratios, with n = 87 (6.5%) exceptional findings regarding their isotopic ratios. An additional n = 164 samples were investigated because of elevated dehydroepiandrosterone concentrations, with n = 2 (1.2%) exceptional findings. The remainder were subjected to isotope ratio analysis because of elevated androsterone levels or because this was requested by sports federations.Significant differences between female and male samples were found for the (13)C/(12)C ratios of androsterone and 11beta-hydroxyandrosterone but not for samples taken in or out of competition.A further n = 645 samples originating from other World Anti-Doping Agency accredited laboratories, mainly throughout Europe as well as South America, South Africa and Southeast Asia, were investigated. The (13)C/(12)C ratios of the urinary steroids differ significantly for each geographical region, reflecting the dietary status of the individuals.The system stability over time has been tested by repeated injections of a standard solution and repeated processing of frozen stored blank urine. Despite a drift over time in absolute (13)C/(12)C ratios, no significant change in the difference of (13)C/(12)C (11beta-hydroxyandrosterone) minus (13)C/(12)C (androsterone) could be observed., (Copyright 2009 John Wiley & Sons, Ltd. Copyright 2009 John Wiley & Sons, Ltd.)

The aim of this study was to investigate the possible existence and magnitude of stable carbon isotope discrimination by human 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR). The catalytic portion of HMGR was expressed and purified. The reaction product mevalonate was lactonized and extracted from the reaction mixture by a solid-phase extraction protocol. Stable carbon isotope ratios of mevalonolactone (MVL) were analyzed by gas chromatography-combustion-isotope ratio mass spectrometry. An average fractionation factor (12)k/(13)k of 1.0031 +/- 0.0004 for all carbon atoms contained in MVL was estimated by the method of internal competition. The value was calculated by nonlinear curve fitting, where the ratio (13)C/(12)C of MVL was plotted versus the fraction of reaction.