Title: Doping - Awareness of female tennis players from the world elite group regarding the question of doping. Thesis objective: The objective of this thesis is to gather and evaluate information acquired using questionnaire surveys about the knowledge of top female tennis players from the world elite group related to the issue of doping, and the selection of information and materials for subsequent analysis. Methods: To achieve the objectives of the thesis, the author applied the questionnaire and document analysis sociological research method. Results: After evaluating the results of the questionnaire survey, a significantly insufficient awareness was discovered for one third of the respondents. The remaining two thirds show average knowledge in the monitored area. Key words: doping, anti-doping policy, prohibited list, doping control, tennis. 4

Title: Anti - doping policy and its global development Thesis objective: The main objective of this work is to develop a comprehensive set of anti-doping policy, putting the key milestones in the development and evaluation of importance for the whole process to the anti-doping policy. Methods: The thesis is based on descriptive analysis and document examination. Outcomes: This thesis is intended to provide an overall summary of the global anti-doping policy. Key words: Anti - doping policy - doping - doping control - doping conference

Title Positive findings in doping of athletes in the Czech Republic Objectives The aim of this thesis, based on the doping control protocols from 1993 - 2011 provided by the Czech Anti-Doping Committee, is to assess the most commonly abused doping substances and evaluate sports in which doping is most frequently abused, and attitudes of athletes who have violated doping regulations. Methods The main method used in this thesis was an analysis of documents, especially doping protocols, which led to gathering important data and compile final tables, which were subsequently processed and evaluated. Finally, aninterview with two bodybuilders who have received completely different penalties for the same offense. Results This thesis comes to the conclusion that the most commonly used substances are, in order: testosterone, nandrolone, marijuana, ephedrine and then metandienon. The most common individual sports, where the use of doping substances is most commonly encoutered include bodybuilding and powerlifting. In team sports, it is hockey, rugby, water polo and handball. Use is most prevalent in men, and less common in women. Keywords Anti-doping policy, doping, doping control, doping substance, sport.

Sport as a kind of physical activity should, above all, be a way of entertainment for people. However, one cannot forget to mention the other aspects, which sport provides to people, whether from the point of view of health or psychology. It particularly concerns the improvement of both physical and mental form, prevention against health problems, relaxation, improvement of communication, cooperation etc. Sport has become a phenomena of these days. Shining careers, tremendous popularity and last but not least sometimes huge amounts of money offered to the best sportsmen lead to swindles as a matter of course. Nowadays, the desire for success can overcome the limits of fair play, and therefore a lot of sportsmen tend to the use of illegal aids in order to have an advantage over their rivals. In spite of that, doping is not the term only connected with top-performance sports, but also with recreation sports. Dope substances are used on all sporting levels at the present. The main aim of this work is to introduce readers to the problems of using dopes and their detection. The work describes the ways in which doping checks proceed, and in which sports and on which levels doping checks can be anticipated. Keywords Doping, blood tests, testing, doping controls , prohibited substances , prohibited metods...

This thesis describes the in vitro production of drug metabolites using fungi of the Cunninghamella species. The metabolites were characterized with mainly liquid chromatography-mass spectrometry using ion-trap and quadrupole-time-of-flight instruments. A fungal in vitro model has several advantages e.g., it is easily up-scaled and ethical problems associated with animal-based models are avoided. The metabolism of bupivacaine and the selective androgen receptor modulators (SARMs) S1, S4 and S24 by the fungi Cunninghamella elegans and Cunninghamella blakesleeana was investigated. The detected metabolites were compared with those formed in vitro and in vivo by human and horse and most phase I metabolites formed by mammals were also formed by the fungi. The higher levels of bupivacaine metabolites in the fungal samples allowed an extensive mass spectrometric structural characterization which shows that the fungi are relevant metabolic models. Glucuronides are important drug metabolites but they are difficult to synthesize. The discovery that the fungus Cunninghamella elegans formed large amounts of glucosides led to the idea that they could be used to form glucuronides. A new concept was developed where a fungal incubate containing a SARM S1 glucoside was mixed with the free radical tetramethylpiperidinyl-1-oxy (TEMPO), sodium bromide and sodium hypochlorite which produced a glucuronide. Isolation and characterization by nuclear magnetic resonance spectroscopy proved that the new method could produce glucuronides for use as reference material. An investigation of reactive metabolite formation of the drugs paracetamol, mefenamic acid and diclofenac by the fungus Cunninghamella elegans was performed. It was demonstrated for the first time that the fungus could produce glutathione, glutathione ethyl-ester, cysteine and N-acetylcysteine conjugates that are indicative of a preceding formation of reactive intermediates. A comparison with conjugates formed by human liver microsomes showed that both models formed identical metabolites. The presented investigations prove that Cunninghamella fungi are relevant drug metabolism models. They show that the fungi to a large extent forms the same metabolites as mammals and that they can produce metabolites for use as reference material in, e.g. doping control. It was also demonstrated that the fungal model can be used in the important assessment of drug toxicity.

Made available in DSpace on 2014-08-20T13:32:48Z (GMT). No. of bitstreams: 1 tese_thais_farias_collares.pdf: 1094762 bytes, checksum: 647dd933472bf9127c0164ee875617ea (MD5) Previous issue date: 2013-10-17 / The history of sports competition has been related to the use of methods for physical training associated with physiological methods to increase athlete performance. Ethical issues involving the high performance sport and doping are confused with the history of competitive sports. The World Anti - Doping Agency (WADA) prohibits the use of substances or methods to artificially increase sports performance. Currently, 258 substances are listed by WADA and athletes around the world are subjected to tests proving the non-use of doping. EPO is a glycoprotein hormone that has as its main physiological effect induction of erythropoiesis and thereby improving the capacity to transport oxygen in the blood. For these reason EPO has been included in WADA list. The analytical differentiation of endogenous erythropoietin from its recombinant counterpart, using isoelectric focusing and double blotting is a milestone in the detection of doping with recombinant erythropoietin. However, several analogs of the original recombinant EPO are not easily detectable by standard IEF method, requiring the development of alternatives for the detection of doping. In order to improve the current methods of EPO detection, monoclonal and polyclonal antibodies against EPO were obtained and used in various techniques for detection of EPO in biological samples. However, the specificity of these antibodies has been quite controversial and discussed. Therefore, it is necessary to obtain antibodies capable of reacting specifically with the EPO. In this study, rHuEPO was inoculated in New Zealand rabbits to generate a polyclonal antibody (pAb anti-rHuEPO). The pAb was characterized for its potential in detecting rHuEPO using different approaches. The pAb anti- rHuEPO identified the expression of recombinant protein in eukaryotic cells and was able to detect rHuEPO in a suspension at 0.1 μg/mL, showing its potential as a tool for detection of doping by rHuEPO. / A história da competição esportiva sempre esteve relacionada à utilização de metodologias de treinamento físico associadas a métodos de incremento fisiológico do atleta, visando sua máxima performance. Questões éticas envolvendo o esporte de alto rendimento e o doping se confundem com a própria história do esporte competitivo. A Agência Mundial Anti-Doping (WADA) proíbe o uso de substâncias ou métodos capazes de aumentar artificialmente o desempenho esportivo. Atualmente, 258 substâncias estão na lista da WADA e atletas do mundo inteiro são submetidos a testes comprobatórios da não utilização do doping. Entre essas substâncias ilícitas para uso por atletas destaca-se a eritropoetina (EPO). A EPO é um hormônio glicoproteico que possui como principal efeito fisiológico a indução da eritropoiese e consequente melhoria da capacidade de transporte de oxigênio no sangue. A diferenciação analítica da eritropoetina endógena produzida a partir de sua contraparte recombinante usando focalização isoelétrica e duplo blotting é um marco na detecção do doping com eritropoetina recombinante. Anticorpos monoclonais e policlonais específicos anti-EPO foram obtidos e utilizados em várias técnicas para a detecção da EPO em amostras biológicas a fim de melhorar os métodos atuais de detecção. Contudo, a especificidade destes anticorpos tem sido bastante controversa e discutida. Portanto, a obtenção de anticorpos específicos capazes de reagir especificamente com a EPO faz-se necessária. Neste estudo, a rHuEPO foi utilizada para imunizar coelhos New Zealand para gerar um anticorpo policlonal (pAb anti-rHuEPO). O pAb foi caracterizado quanto ao seu potencial na detecção da rHuEPO usando diferentes metodologias. O pAb anti-rHuEPO identificou a expressão da proteína recombinante em células eucarióticas e foi capaz de detectar rHuEPO em suspensão até 0,1 μg/mL, comprovando seu potencial como ferramenta para detecção do doping por rHuEPO.

Peer reviewed literature currently provides little knowledge of how to recruit and train Doping Control Officers (DCOs) for Mega Sport Events. This study adds to the existing body of literature on creating knowledge transfer and legacy for future Mega Event Organizing Committees. A phenomenological designed study was conducted through the use of a qualitative on-line survey during the 2015 Toronto Pan American Games. A survey response rate of 58% was achieved. Survey responses by DCOs were analyzed using inductive coding to identify four main themes; i) DCO motivations, ii) training program design, iii) engagement and support, and iv) barriers to participation in the results. Recommendations include using mixed methods for future studies, provide DCOs with training and evaluation opportunities prior to the Mega Sport Event, and developing mentorship opportunities during games operations. Additional research is needed to create greater understanding of DCO motivations and perceptions beyond this study.

El di(2-etilhexil) ftalat (DEHP) és un plastificant que s’afegeix als productes de clorur de polivinil (PVC) per a dotar-los de més flexibilitat. El material mèdic fet de PVC, i en particular els dispositius i bosses que s’utilitzen en les transfusions de sang, conté el DEHP com additiu. Així, el receptor d’una transfusió està altament exposat a aquest compost. L’objectiu de la tesi va ser estudiar els metabòlits del DEHP en orina com a possibles marcadors de la pràctica d’una transfusió de sang en l’esport. Es va desenvolupar i validar un mètode d’anàlisi per cromatografia líquida acoblada a espectrometria de masses en tàndem per a la quantificació dels principals metabòlits del DEHP en orina humana: mono-(2-etilhexil) ftalat (MEHP), mono-(2-etil-5-hidroxihexil) ftalat (MEHHP), mono-(2-etil-5-oxohexil) ftalat (MEOHP), mono-(2-carboximetilhexil) ftalat (2cx-MMHP) i mono-(2-etil-5-carboxipentil) ftalat (5cx-MEPP). El mètode es va aplicar a mostres procedents de voluntaris sans (grup control), de pacients hospitalitzats que havien rebut una transfusió de sang i de pacients hospitalitzats sotmesos a tractaments mèdics amb materials de PVC i no a transfusions. Es van obtenir diferències significatives en les concentracions dels tres metabòlits estudiats (MEHP, MEHHP, MEOHP) entre les mostres dels pacients transfosos respecte els altres dos grups de població. El mètode també es va aplicar a mostres d’orina de vint-i-cinc voluntaris sans que s’havien sotmès a un procediment d’autotransfusió. Els resultats van indicar concentracions elevades dels cinc metabòlits del DEHP en orina fins a les 48 hores després d’haver rebut la sang. Finalment, es van determinar les concentracions dels cinc metabòlits de DEHP en una població d’esportistes i es van calcular límits de referència que permetessin sospitar d’una transfusió. Així doncs, els resultats indiquen que la mesura dels metabòlits de DEHP en orina pot ser usada com una eina pel cribatge de l’ús de transfusions en l’esport. / The plasticizer di(2-ethylhexyl)phthalate (DEHP) is used in polyvinyl chloride products (PVC) to increase its flexibility. Medical devices made of PVC, especially blood bags used in blood transfusions, contain DEHP as additive. Therefore, subjects submitted to blood transfusion are widely exposed to this compound. The aim of the project was to evaluate DEHP metabolites in urine as possible markers of the use of a blood transfusion in sports. An analytical method was developed and validated to quantify the main DEHP metabolites mono-(2-ethylhexyl)phthalate (MEHP), mono-(2-ethyl-5-hydroxyhexyl)phthalate (MEHHP), mono-(2-ethyl-5-oxohexyl)phthalate (MEOHP), mono-(2-carboxymethylhexyl)phthalate (2cx-MMHP) and mono-(2-ethyl-5-carboxypentyl)phthalate (5cx-MEPP), in human urine by liquid chromatography tandem mass spectrometry. The methodology was applied to samples belonging to healthy volunteers (control group), hospitalized patients subjected to blood transfusions and hospitalized patients subjected to medical treatments involving plastic material different to blood transfusions. Significant differences were obtained in the concentrations of the three metabolites studied (MEHP, MEHHP, MEOHP) between transfused patients samples’ and the other two population groups. The method was also applied to urine samples from twenty-five healthy volunteers who were subjected to an autologous blood transfusion. The results indicated high concentrations of the five DEHP metabolites in urine up to 48 hours after the blood transfusion. Finally, the concentration of the five DEHP metabolites were evaluated in a sportsmen population and reference limits to allow suspicion of blood transfusion were calculated. Thus, the results indicate that the DEHP metabolites could be used as markers of blood transfusions in sports.