Drug Use on Mont Blanc: A Study Using Automated Urine Collection.

Mont Blanc, the summit of Western Europe, is a popular but demanding high-altitude ascent. Drug use is thought to be widespread among climbers attempting this summit, not only to prevent altitude illnesses, but also to boost physical and/or psychological capacities. This practice may be unsafe in this remote alpine environment.

Improved Steroids Detection and Evidence for Their Regiospecific Decompositions Using Anion Attachment Mass Spectrometry.

Nonpolar anabolic steroids are doping agents that typically do not provide strong signals by electrospray ionization-mass spectrometry (ESI-MS) owing especially to the low polarity of the functional groups present. We have investigated the addition of anions, in ammonium salt form, to anabolic steroid samples as ionization enhancers and have confirmed that lower instrumental limits of detection (as low as 10 ng/mL for fluoxymesterone-M) are obtained by fluoride anion attachment mass spectrometry, as compared to ESI(+)/(-) or atmospheric pressure photoionization (APPI)(+). Moreover, collision-induced decomposition (CID) spectra of precursor fluoride adducts of the bifunctional steroid "reduced pregnenolone" (containing two hydroxyl groups) and its d4-analogue provide evidence of regiospecific decompositions after attachment of fluoride anion to a specific hydroxyl group of the steroid.

Formation of the diuretic chlorazanil from the antimalarial drug proguanil--implications for sports drug testing.

Chlorazanil (Ordipan, N-(4-chlorophenyl)-1,3,5-triazine-2,4-diamine) is a diuretic agent and as such prohibited in sport according to the regulations of the World Anti-Doping Agency (WADA). Despite its introduction into clinical practice in the late 1950s, the worldwide very first two adverse analytical findings were registered only in 2014, being motive for an in-depth investigation of these cases. Both individuals denied the intake of the drug; however, the athletes did declare the use of the antimalarial prophylactic agent proguanil due to temporary residences in African countries.

Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures.

Nonspecific interactions between blotted proteins and unrelated secondary antibodies generate false positives in immunoblotting techniques. Some procedures have been developed to reduce this adsorption, but they may work in specific applications and be ineffective in others. "Double-blotting" has been developed to overcome this problem.

Detection of tetracosactide in plasma by enzyme-linked immunosorbent assay (ELISA).

As a synthetic analogue of adrenocorticotropic hormone (ACTH), tetracosactide is prohibited in sport by the World Anti-Doping Agency (WADA). An enzyme-linked immunosorbent assay (ELISA) method is proposed for detection of this drug in plasma. Since its structure corresponds to the 24 N-terminal of the 39 amino acids of the natural endogenous peptide ACTH, tetracosactide can be detected with a commercial ELISA kit for ACTH that uses antibodies, the epitopes of which are located in the 1-24 part of ACTH.

Ergogenic and metabolic effects of oral glucocorticoid intake during repeated bouts of high-intensity exercise.

All systemically administered glucocorticoids (GC) are prohibited in-competition, because of the potential ergogenic effects. Although short-term GC intake has been shown to improve performance during submaximal exercise, literature on its impact during brief intense exercise appears to be very scant. The purpose of this study was to examine the ergogenic and metabolic effects of prednisone during repeated bouts of high-intensity exercise.

Acute apnea swimming: metabolic responses and performance.

Competitive swimmers regularly perform apnea series with or without fins as part of their training, but the ergogenic and metabolic repercussions of acute and chronic apnea have not been examined. Therefore, we aimed to investigate the cardiovascular, lactate, arterial oxygen saturation and hormonal responses to acute apnea in relation to performance in male swimmers. According to a randomized protocol, 15 national or regional competitive swimmers were monitored while performing four 100-m freestyle trials, each consisting of four 25-m segments with departure every 30 seconds at maximal speed in the following conditions: with normal frequency breathing with fins (F) and without fins (S) and with complete apnea for the four 25-m segments with (FAp) and without fins (SAp).

Data-handling strategies for metabonomic studies: example of the UHPLC-ESI/ToF urinary signature of tetrahydrocannabinol in humans.

Metabonomics has become a very valuable tool and many research fields rely on results coming out from this combination of analytical techniques, chemometric strategies, and biological interpretation. Moreover, the matrices are more and more complex and the implications of the results are often of major importance. In this context, the need for pertinent validation strategies comes naturally.

The problem of anti-doping control of luteinizing hormone in boxing.

Luteinizing hormone (LH) is physiologically produced by the anterior pituitary gland. Male athletes may use pharmaceutical LH for doping since it increases the production of testosterone by testes. This hormone is thus on the World Anti-Doping Agency (WADA) list of substances prohibited for males.

Changes in steroid hormones during an international powerlifting competition.

The purpose of this study was to assess changes in the steroid hormone levels of elite athletes during an international powerlifting competition. Baseline cortisol, DHEA and testosterone were determined in saliva samples in 19 (8 men, 11 women) junior and sub-junior athletes on the day before competition, and then on the competition day during the official weighing and in the hour after competition. Performance was determined by total output and the Wilks formula.

Detection of erythropoiesis-stimulating agents in human anti-doping control: past, present and future.

Stimulation of erythropoiesis is one of the most efficient ways of doping. This type of doping is advantageous for aerobic physical exercise and of particular interest to endurance athletes. Erythropoiesis, which takes place in bone marrow, is under the control of EPO, a hormone secreted primarily by the kidneys when the arterial oxygen tension decreases.

Cave digitabula!

We report here observations clearly demonstrating that gloves can have a deleterious impact on an IEF experiment. These observations were made during the practice of analyses for anti-doping control of erythropoietin. We show that the wearing of nitrile gloves to apply the catholyte strip onto the IEF gel may be responsible for dramatic distortions in the pattern of this hormone.

Simultaneous quantification and qualification of synacthen in plasma.

Tetracosactide (Synacthen), a synthetic analogue of adrenocorticotropic hormone (ACTH), can be used as a doping agent to increase the secretion of glucocorticoids by adrenal glands. The only published method for anti-doping control of this drug in plasma relies on purification by immunoaffinity chromatography and LC/MS/MS analysis. Its limit of detection is 300 pg/mL, which corresponds to the peak value observed 12 h after 1 mg Synacthen IM administration.

Effects of exercise on the isoelectric patterns of erythropoietin.

Objectives: Recombinant erythropoietin has a strong impact on aerobic power and is therefore one of the most potent doping agents in endurance sports. The anti-doping control of this synthetic hormone relies on the detection, in the urine, of its isoelectric pattern, which differs from that of the corresponding natural hormone, the latter being typically more acidic than the former. However, a small number of natural urinary patterns, referred to as "atypical patterns," are less acidic than the dominant form.

Biosafety in ex vivo gene therapy and conditional ablation of lentivirally transduced hepatocytes in nonhuman primates.

Ex vivo gene therapy is an interesting alternative to orthotopic liver transplantation (OLT) for treating metabolic liver diseases. In this study, we investigated its efficacy and biosafety in nonhuman primates. Hepatocytes isolated from liver lobectomy were transduced in suspension with a bicistronic liver-specific lentiviral vector and immediately autotransplanted (SLIT) into three cynomolgus monkeys.

Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures.

Nonspecific interactions between blotted proteins and unrelated secondary antibodies generate false positives in immunoblotting techniques. Some procedures have been developed to reduce this adsorption but they may work in specific applications and be ineffective in other ones. "Double-blotting" has been developed to overcome this problem.

Caspase activation, sialidase release and changes in sialylation pattern of recombinant human erythropoietin produced by CHO cells in batch and fed-batch cultures.

The activation of caspases represents a crucial turning point during a batch or a fed-batch culture of mammalian cells. It not only affects the quantity but also the quality of the recombinant glycoprotein produced. In this study, the activation of various caspases, the release of intracellular sialidase and the changes in sialylation pattern of a recombinant product, erythropoietin (EPO), in the culture medium were analyzed in both batch and fed-batch cultures.

Isoelectric profiles of human erythropoietin are different in serum and urine.

By adding a step of immunoaffinity to the method we had previously developed for analysing erythropoietin (EPO) in urine, we were able to study the isoelectric profiles of this hormone in human serum samples. This method was sensitive enough to investigate samples presenting physiological levels of this hormone. Comparison with the corresponding profiles in urine showed that natural EPO was systematically more acidic in urine.

Detection of recombinant human erythropoietin in urine for doping analysis: interpretation of isoelectric profiles by discriminant analysis.

The detection in urine of recombinant human erythropoietin (rHuEPO), a hormone misused by endurance athletes as a doping agent, is based on the differentiation of its isoelectric pattern from that of the corresponding natural hormone. Different empirical criteria have been proposed for discriminating the images of the patterns but none of them have been elaborated from a rational statistical approach. Discriminant analysis was applied to a dataset of profiles defined as positive (116 profiles from 26 subjects) (presence of rHuEPO and possibly residual natural endogenous hormone) and negative (131 profiles from 131 subjects) (presence of natural endogenous hormone only).