Projets de recherche

Code: 241A04MT

Already in 2006 the synthesis of (17α,20E)‐17,20‐[(1‐methoxyethylidene) bis (oxy)]‐3‐oxo‐19‐norpregna‐4,20‐diene‐21‐carboxylic acid methyl ester (YK-11) was published and investigations into its pharmacological properties demonstrated the potential of YK-11 as a selective androgen receptor modulator (SARM). Albeit YK-11 has not been approved for humans until now, it was first detected in a black-market product in 2017 and several times thereafter. According to the Dietary Supplement Label Database issued by the National Institutes of Health, at least 6 different preparations are currently available in the United States alone claiming to contain YK-11.

In 2018 the human metabolism of YK-11 was investigated and two main urinary metabolites were detected suitable to be implemented into routine doping control procedures. YK-11 itself was not detected in urine, presumably due to the fact that it undergoes fast metabolism in humans. As the parent compound YK-11 will not be found in urine after administration, doping controls can only rely on the identified urinary metabolites. Both metabolites were synthesized in order to elucidate their chemical structure, but no reference material at a larger scale was synthesized at that time suitable to be provided to all doping control laboratories worldwide.

In order to enable all laboratories to implement the urinary metabolites of YK-11 into their screening procedures, the production of a suitable reference material of both YK-11 metabolites will be inevitable. Therefore, the aim of this research project is to reinvestigate the different potential routes of chemical synthesis, optimize these and produce reference material of both YK-11 metabolites in order to distribute this to all accredited doping control laboratories.

Code: 241A02MT

In sports drug testing, isotope ratio mass spectrometry is employed to enable the differentiation between endogenous and exogenous sources of urinary steroids, i.e. steroids produced inside the body or administered. Testosterone or testosterone-prohormones may be used by athlete for performance enhancement. As these steroids are endogenous, their presence alone in human urine is not suspicious. Only if urinary concentrations may exceed individual thresholds, a confirmation of the exogenous source becomes necessary and carbon isotope ratios proved to be the method of choice here.

Sample preparation protocols for isotope ratio mass spectrometry-based method are complicated and time-consuming as the purity of analytes is crucial and a prerequisite for valid isotope ratio determinations. Currently applied sample preparations encompass solid and liquid-liquid extraction steps to pre-purify and concentrate urine samples prior to the final clean-up step employing high performance liquid chromatography and fraction collection. This powerful tool for sample clean-up comes along with relatively long run-times per sample of more than 45 min and it may be necessary to use even a two-fold separation here.

This research project aims for improving this crucial step in sample preparation and shorten the run-times per sample by employing supercritical fluid chromatography coupled to a novel fraction collection tool. In supercritical fluid chromatography, pressurized carbon dioxide is used as solvent offering unique features for chromatographic separation not directly comparable to the commonly applied mixtures of water and organic solvent. Therefore, a novel sample preparation method suitable for isotope ratio mass spectrometry will be developed and validated within this project investigating the benefits of supercritical fluid chromatography in sports drug testing. To evaluate the new method and facilitate its implementation into routine doping controls, reference population-based values will be determined and compared to existing routine applications.

The aim of the project is to develop an anti-doping education framework for the World ParaVolley Association (WPV) For a complete list of National Federations involved, refer to World ParaVolley’s membership.

The proposed objectives will be achieved through a mixedmethods approach delineating action-oriented research. The mixed methods approach will be crucial to understanding the sports person's experience with NADA’s anti-doping educational awareness interventions and, thereby, the measures to improve them.

This project investigates the beliefs, attitudes, and awareness of doping among student-athletes in Singapore. A mixed methodologies of quantitative online surveys and qualitative interviews, focusing on student-athletes attending the National University of Singapore (NUS). In hopes to offer insight into Singapore student-athletes’ doping vulnerability, experience of the anti-doping system, and the factors shaping these aspects. Additionally, shedding light on nonsport influences on substance use among young people in larger society and how these influences might indirectly affect student-athletes’ doping attitudes in sport.

The proposed project will identify and characterize sportspecific stressors contributing to doping vulnerability among athletes, to include exploring their impact, frequency, severity, timing, and duration. Additionally, it seeks to investigate how psychological flexibility moderates the relationship between these stressors and doping vulnerability, thereby protecting athletes from doping.

A qualitative approach and informed by a co-design process with national anti-doping and sporting organizations. This project seeks to interview women in three countries to 1) investigate the role of support personnel (e.g., coaches, trainers, partners) in facilitating or preventing PIED use, and 2) to understand the broader psycho-socio-cultural factors that contribute to facilitating or preventing PIED use among female athletes. Investigating when, why, and how women choose to use or avoid PIEDs this project will gain important insights on women’s lived experiences with PIEDs.

A cross-sectional survey design and recruit a large sample of elite athletes (1500) from the said countries. The survey will include a number of validated screening tools in Spanish about self-compassion, mindfulness, sport anxiety, perfectionism, and doping susceptibility. Participants will also be invited to take part in a podcast series and to share their insights and experiences on doping in sport. In order to contribute towards more informed decision-making playing a crucial role in the prevention of doping behavior in the PANRADO Region, while focusing on a positive psychology approach emphasizing the role of mindfulness and selfcompassion.

Code: 23GD03HS

Gene doping is defined as “the non-therapeutic use of genes, genetic elements and/or modified cells that have the capacity to enhance athletic performance” and is categorized as a threat to the integrity of sport and the health of athletes by the World Anti-Doping Agency (WADA). The only direct approaches for detecting gene doping are PCR assays, so introducing a novel alternative approach for rapid and accurate identification of gene doping is vital. In this project, a novel ultrasensitive detection platform will be designed based on the combination of 2 different technologies i.e. CRISPR and Microfluidics. CRISPR/CAS systems can survey the whole genome and detect transgenic DNA (tDNA) using a single-stranded guide RNA (gRNA). In the presence of a target gene, a ternary complex (Cas12/gRNA/tDNA) will be formed and the quenched fluorescence probe will be degraded by collateral cleavage activity of the ternary complex. Microfluidics fluorescence detection can provide exquisite sensitivity. The combination of CRISPR/CAS as a biological gene doping detection system and a microfluidics system can provide multiplex gene doping detection (up to 96 samples with 96 detection assays per chip). Pre-amplification technologies such as recombinase polymerase amplification will be applied in order to reach single molecule (1 aM) sensitivity. Here, we will investigate more than 20 genes including CSH1, CSH1,CSH2, CSHL1, EPO, GDFB, GH1, GH2, IGF1, IGF2, MYOG, PPARd, VEGF, etc. as a proof of concept. However, the number of assay combinations could be 96 samples with 96 detection assays (96 different Cas/gRNAs). The gRNAs will be designed to target exon-exon junctions, specific sequences of viral vectors, such as CMVp (cytomegalovirus promoter), TkpA (thymidine kinase polyA from herpes simplex virus), hexon (major virus capsid protein from adenovirus), and CMVp (cytomegalovirus promoter) etc. The use of combinations of different gRNAs for the same gene (several gRNAs for different exon-exon junctions) with virus targeted-gRNAs will allow us to detect doping genes and different vectors simultaneously and dramatically increase the successful detection of tDNAs. Moreover, specific CRISPR-reference materials (C-RM) containing modified transgenic sequences will be designed to avoid false positive results due to cross-contamination. Based on the WADA guidelines for gene doping, appropriate no template controls (NTC) and positive template controls (PTC) will be used to determine the specificity, efficiency, sensitivity and limit of detection of the assay. The performance efficiency of the detection platforms will be tested in spiked blood samples. These multiplexed gene doping detection CRISPR-equipped microfluidic platforms could be an ideal and rapidly deployable detection approach, which is accurate, inexpensive and rapid, allowing for anti-gene doping assays in WADA-accredited laboratories.

Code: 23A20MP

GC-C-IRMS is used to confirm the exogenous origin of endogenous anabolic steroids. Samples are analyzed with a fast initial testing procedure to isolate suspicious samples and, afterwards, the suspicious samples are analyzed with an IRMS confirmatory method. High throughput IRMS will allow a substantial expansion of CIR analyses, enabling a considerable increase in doping violation detections and the set-up of an efficient CIR ABP module. High throughput IRMS is also highly desirable in those cases where a large number of samples needs to be tested on IRMS in relative short period of time e.g., preseason/regular season testing and Olympic Games.