NOTEWORTHY ARTICLES 2005

Doping Journal 'Noteworthy' section alerts interested readers about the selected noteworthy original research and selected review articles, book reviews, and meeting reports (published in other journals) on the subject of the Doping Journal scope.

The reference to each article may be accompanied by the referee name (a member of the journal editorial board or a journal reader), the authors' key note comments, the date of the 'noteworthy' alert, letter to the editor link, and links to related articles (if any).

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DJ Noteworthy pages readership 1 September 2004 - 31 August 2005: 4129

Tetrahydrogestrinone: the discovery of a designer steroid
TC Malvey and TD Armsey Ii
Curr Sports Med Rep (1 August 2005) 4(4): 227
[PubMed] [Authors contact]

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Erythropoietin administration does not influence the GH-IGF axis or makers of bone turnover in recreational athletes
AE Nelson, CJ Howe, TV Nguyen, MJ Seibel, RC Baxter, DJ Handelsman, R Kazlauskas, and KK Ho
Clin Endocrinol (Oxf) (1 September 2005) 33(3): 305
[PubMed] [Authors contact]

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Quantitative analysis of images in erythropoietin doping control
I Bajla, I Hollander, G Gmeiner, and Ch Reichel
Med Biol Eng Comput (1 May 2005) 43(3): 403
[PubMed] [Authors contact]

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Androgenic-anabolic steroids abuse in males
L Di Luigi, F Romanelli, and A Lenzi
J Endocrinol Invest (1 January 2005) 28(3 Suppl): 81
[PubMed] [Authors contact]

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Growth hormone and connective tissue in exercise
S Doessing and M Kjaer
Scand J Med Sci Sports (1 August 2005) 15(4): 202
[PubMed] [Authors contact]

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Accidental breaches of the doping regulations in sport: is there a need to improve the education of sportspeople?
SJ Somerville, M Lewis, and H Kuipers
Br J Sports Med (1 August 2005) 39(8): 512
[PubMed] [Authors contact]

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Erythropoietin in Sports: A New Look at An Old Problem
Joshua Scott and George C Phillips
Toxicol Lett (1 August 2005)
[PubMed] [Authors contact]

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Cannabis use to enhance sportive and non-sportive performances among French sport students
FO Lorente, P Peretti-Watel, and L Grelot
Addict Behav (1 August 2005) 30(7): 1382
[PubMed] [Authors contact]

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Challenges in Detecting the Abuse of Growth Hormone in Sports
Cathy M McHugh, Roderick T Park, Peter H Sonksen, and Richard I G Holt
Clin Chem (14 July 2005)
[PubMed] [Authors contact]

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Haematological testing to fight blood doping response - letter to the editors: haematological testing and antidoping policies - re: Robinson N, Schattenberg L, Zorzoli M, Mangin P, Saugy M. Haematological analysis conducted at the departure of the tour de france 2001
N Robinson, P Mangin, and M Saugy
Int J Sports Med (1 July 2002) 26(6): 510
[PubMed] [Authors contact]

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Adverse effects of anabolic steroids in athletes A constant threat
C Maravelias, A Dona, M Stefanidou, and C Spiliopoulou
Toxicol Lett (5 July 2005)
[PubMed] [Authors contact]

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Forecasting of top athletic performance
P Derevenco, M Albu, and E Duma
Rom J Physiol (1 January 2002) 39-40: 57
[PubMed] [Authors contact]

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The negative regulation of red cell mass by neocytolysis: physiologic and pathophysiologic manifestations
L Rice and C Alfrey
Cell Physiol Biochem (1 January 2005) 15(6): 245
[PubMed] [Authors contact]

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Herbal supplements in sport: use and abuse
Joshua Scott and George C Phillips
Ann Ist Super Sanita (1 January 2005) 41(1): 35
[PubMed] [Authors contact]

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Adverse effects of anabolic steroids in athletes A constant threat
C Maravelias, A Dona, M Stefanidou, and C Spiliopoulou
Toxicol Lett (5 July 2005)
[PubMed] [Authors contact]

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Medication and supplement use by athletes
Ciocca M
Clin Sports Med (1 July 2005) 24(3): 719
[PubMed] [Authors contact]

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Chimera on a Bike?
Credit: Stein W
Science (24 June 2005) 308(5730): 1864b
[PubMed] [Related Doping J News: 1 | 2 | 3 | 4 | 5 | 6 ] [Authors contact]

Excerpt: "...Last September, Olympian Tyler Hamilton, 34, of Boulder, Colorado, was accused of taking a blood transfusion to boost his performance after a newly developed test showed he had two different types of red blood cells. Hamilton denied the charge, and with the help of geneticist David Housman of the Massachusetts Institute of Technology in Cambridge, he has been arguing that he might be a chimera: an organism with a mix of genetically distinct cells. Human chimeras are not all that rare, Housman told a board arbitrating the case in March. Mothers and fetuses often exchange blood-producing stem cells, and fetuses can also get foreign cells from sharing the womb with a "vanishing twin," he said. But the arbitrators didn't bite, voting 2 to 1 to uphold a 2-year suspension and stating that blood doping was "the only reasonable conclusion." Encouraged by the split decision, however, Hamilton is again appealing, this time to a sport arbitration court in Switzerland... Mother-fetus chimerism is unlikely to produce foreign cells at the levels found in Hamilton's blood - about 2% - says geneticist Wendy Robinson of the University of British Columbia in Vancouver, Canada. But Housman predicts that more athletes who are actually chimeras will show up as suspected blood dopers. And, wrote Hamilton this month in his online journal, "If we've accomplished nothing else in this case, we have put a spotlight on the vanishing twin phenomenon.""

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Governing Doped Bodies: The World Anti-Doping Agency and the Global Culture of Surveillance
Jin-Kyung Park
University of Illinois - Urbana-Champaign
Cultural Studies <=> Critical Methodologies (2005) 5(2): 174-188
[PubMed] [Authors contact]

Abstract: "This essay examines the governing practices of the World Anti-Doping Agency (WADA), an organization established in 1999 to cope with the crisis of illicit performance-enhancing drug use in international sport. The background, structure, and policies of WADA are analyzed while reflecting upon recent cultural studies debates on governmentality. In doing so, it is shown how WADA policies fundamentally work to police athletic bodies. Also demonstrated is that WADA embodies a First World, technology-driven governance of doping. Key Words: governmentality, surveillance, body, doping, globalization."

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The world anti-doping code 2003 - consequences for physicians associated with elite athletes
Striegel H, Rossner D, Simon P, and Niess AM
Medical Clinic and Policlinic, Department of Sports Medicine, University of Tubingen, Silcherstrasse 5, 72076 Tubingen, Germany
Int J Sports Med (1 April 2005) 26(3): 238
[PubMed] [Authors contact]

Abstract: "The purpose of the World Anti-Doping Code 2003 and the 2004 Prohibited List is to create a universal international standard to fight doping in competitive sports. The result of this is a whole series of changes for doctors with regard to their work with competitive athletes. The revised definition of doping now includes physicians in the group of persons who can fulfil the elements of a doping offence. Moreover, the mere possession of substances appearing on the Prohibited List represents a violation of anti-doping regulations. The 2004 Prohibited List includes several changes to the Olympic Movement List from 2003. Caffeine, for example, was removed from the list. Cannabinoids, on the other hand, are now prohibited in competition for all sports. The same is true for all forms of glucocorticosteroids. Therapeutic use exemptions in an abbreviated process are possible for the administration of glucocorticosteroids by non-systemic routes, as well as inhalative therapy with the beta-2-agonists formoterol, salbutamol, salmeterol, and termbutalin. In other cases, a therapeutic use exemption is possible using a standard application process. Further changes will become effective in the 2005 Prohibited List. In 2005, it is essential that beta-2-agonists are prohibited in and out of competition. HCG and LH are prohibited for all athletes. Dermatological preparations of glucocorticosteroids are no longer prohibited, and intravenous infusions will be a prohibited method in 2005, except as a legitimate acute medical treatment. In cases of violations of anti-doping regulations where it is permissible for the affected person to furnish proof of exoneration, the burden of proof is not higher than that required to prove the violation. The sanctions provided for in the World Anti-Doping Code follow a principle of rules and exceptions which at first glance seems difficult to understand. In the case of doping violations by physicians, the anti-doping code provides - as a general rule - for exclusion from sports associations for at least four years. Since several of the changes are questionable under constitutional aspects, it remains to be seen whether the World Anti-Doping Code 2003 will allow the achievement of a universal standard to combat doping."

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Screening for unknown synthetic steroids in human urine by liquid chromatography-tandem mass spectrometry
Thevis M, Geyer H, Mareck U, Schanzer W
Institute of Biochemistry, German Sport University Cologne, Carl-Diem Weg 6, 50933, Cologne, Germany
J Mass Spectrom (2 June 2005) ePub ahead of print
[PubMed] [Related Article] [Authors contact]

Abstract: "Chemically modified steroids (designer steroids), including tetrahydrogestrinone and norbolethone, pose a threat to the integrity of the sport community. These compounds have recently been detected in urine specimens from athletes, resulting in temporary or permanent suspension from amateur and/or professional competition. Triple quadrupole mass spectrometers enable doping control laboratories to screen for unknown, anabolic, androgenic steroids utilizing precursor ion scans. On the basis of common dissociation patterns of steroids with common structural features, characteristic product ions were selected to serve as diagnostic markers for previously unidentified drugs or drug metabolites in human urine samples. An assay was established to complement standard screening procedures. Urine specimens were enzymically hydrolyzed, partitioned into ether, concentrated, and analyzed by precursor ion scanning. Spectra from samples fortified with eight standard compounds (methyltestosterone, ethyltestosterone, 1-testosterone, gestrinone, dihydrogestrinone, tetrahydrogestrinone, norbolethone, and propyltrenbolone) and one deuterium-labeled analog (d(4)-tetrahydrogestrinone) at 50 ng/ml of urine, had precursor ion peaks other than those from common endogenous steroids. Subsequent product ion scan experiments on precursor ions of peaks of unknown origin provided structural identification of the unknown compounds."

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Screening for unknown synthetic steroids in human urine by liquid chromatography-tandem mass spectrometry
Thevis M, Geyer H, Mareck U, Schanzer W
Institute of Biochemistry, German Sport University Cologne, Carl-Diem Weg 6, 50933, Cologne, Germany
J Mass Spectrom (2 June 2005) ePub ahead of print
[PubMed] [Related Article] [Authors contact]

Abstract: "Chemically modified steroids (designer steroids), including tetrahydrogestrinone and norbolethone, pose a threat to the integrity of the sport community. These compounds have recently been detected in urine specimens from athletes, resulting in temporary or permanent suspension from amateur and/or professional competition. Triple quadrupole mass spectrometers enable doping control laboratories to screen for unknown, anabolic, androgenic steroids utilizing precursor ion scans. On the basis of common dissociation patterns of steroids with common structural features, characteristic product ions were selected to serve as diagnostic markers for previously unidentified drugs or drug metabolites in human urine samples. An assay was established to complement standard screening procedures. Urine specimens were enzymically hydrolyzed, partitioned into ether, concentrated, and analyzed by precursor ion scanning. Spectra from samples fortified with eight standard compounds (methyltestosterone, ethyltestosterone, 1-testosterone, gestrinone, dihydrogestrinone, tetrahydrogestrinone, norbolethone, and propyltrenbolone) and one deuterium-labeled analog (d(4)-tetrahydrogestrinone) at 50 ng/ml of urine, had precursor ion peaks other than those from common endogenous steroids. Subsequent product ion scan experiments on precursor ions of peaks of unknown origin provided structural identification of the unknown compounds."

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Oro-facial injuries in Central American and Caribbean sports games: a 20-year experience
Amy E
Department of Physical Medicine, Rehabilitation and Sports Medicine, School of Medicine, University of Puerto Rico, San Juan, Puerto Rico
Dent Traumatol (1 Jun 2005) 21(3): 127
[PubMed] [Authors contact]

Abstract: "Dental services in sports competitions in the Games sponsored by the International Olympic Committee are mandatory. In every Central American, Pan American and Olympic Summer Games, as well as Winter Games, the Organizing Committee has to take all the necessary measures to assure dental services to all competitors. In all Olympic villages, as part of the medical services, a dental clinic is set up to treat any dental emergency that may arise during the Games. Almost every participating country in the Games has its own medical team and some may include a dentist. The major responsibilities of the team dentist as a member of the national sports delegation include: (i) education of the sports delegation about different oral and dental diseases and the illustration of possible problems that athletes or other personnel may encounter during the Games, (ii) adequate training and management of orofacial trauma during the competition, (iii) knowledge about the rules and regulations of the specific sport that the dentist is working, (iv) understanding of the anti-doping control regulations and procedures, (v) necessary skills to fabricate a custom-made and properly fitted mouth guard to all participants in contact or collision sports of the delegation. This study illustrates the dental services and occurrence of orofacial injury at the Central American and Caribbean Sports Games of the Puerto Rican Delegation for the past 20 years. A total of 2107 participants made up the six different delegations at these Games. Of these 279 or 13.2% were seen for different dental conditions. The incidence of acute or emergency orofacial conditions was 18 cases or 6% of the total participants. The most frequent injury was lip contusion with four cases and the sport that experienced more injuries was basketball with three cases."

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Quantitative detection of doping substances by a localised surface plasmon sensor
Kreuzer MP, Quidant R, Badenes G, Marco M-P
Department of Biological Organic Chemistry IIQAB-CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain
Biosens Bioelectron (31 May 2005) ePub ahead of print
[PubMed] [Authors contact]

Abstract: "Within this communication, consistent evidence of a quantitative biosensing principle for steroidal residue analysis is presented. Our approach uses a simple method for the quantitative determination of an anabolic agent called stanozolol (Sz). Sz (Mw 328) is widely used in sports, horse racing and as a growth promoter in animals for human consumption. Through the use of localised surface plasmons (LSPs), sustained by three-dimensional noble metal nano-structures, we have developed a highly specific, label-less immunosensor for the detection of this small organic molecule to low levels (nM range). A main practical advantage over conventional flat extended film surface plasmon resonance (SPR) systems is the simplicity of the optical configuration, since there is no need for cumbersome total internal reflection illumination, thus making integration easier. In addition, the active area of the LSP-based sensor is smaller, decreasing the minimum detectable number of molecules involved in the binding event. Assay times are short and the set-up is comprised of relatively cheap instrumentation. Detection levels found here are comparable with SPR, even at this early stage of development and with further modifications, we envisage sensing down to pM (10(-12)) levels."

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DNA typing: an accessory evidence in doping control
MA Sipoli Marques, LM Pinto Damasceno, HM Gualberto Pereira, CM Caldeira, BF Pereira Dias, D de Giacomo Vargens, ND Amoedo, RO Volkweis, RO Volkweis Viana, FD Rumjanek, and FR Aquino Neto
LABDOP-LADETEC, Instituto de Quimica, Universidade Federal do Rio de Janeiro, Ilha do Fundao, Rio de Janeiro, RJ, Brazil-21949-900
J Forensic Sci (1 May 2005) 50(3): 587
[PubMed] [Authors contact]

Abstract: "A clear positive case for anabolic steroids doping was confounded by alleged urine tampering during doping control procedures. Review of the chain of custody showed no flaws, but nevertheless the athlete was adamant that the urine sample should be analyzed for DNA in order to support her contention that she was not the donor of the sample. The results obtained showed that the urine sample that scored positive for steroids contained nuclear DNA that could not be matched to the DNA obtained from the athlete's blood. On the other hand, the same urine sample contained mitochondrial DNA whose nucleotide sequences spanning the hyper variable regions HV1 and HV2 proved to be identical to those determined in mitochondrial DNA amplified from the athlete's blood. The occurrence of an extraneous genotype is compatible with exogenous nuclear DNA admixture to the athlete's urine. Alternatively, taking in consideration the mitochondrial DNA, we could not exclude that a sibling or a maternal relative of the athlete could have acted as a donor of the urine utilized for doping control and DNA analysis. Both situations point to possible tampering of the urine by the athlete. Adjudication at CAS maintained previous national and international federation decision that there was no proof of a chain of custody flaw to justify the athlete's allegation of urine substitution after collection."

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Qualitative Determination of Synthetic Analogues of Insulin in Human Plasma by Immunoaffinity Purification and Liquid Chromatography-Tandem Mass Spectrometry for Doping Control Purposes
Mario Thevis, Andreas Thomas, Philippe Delahaut, Alain Bosseloir, Wilhelm Schanzer
Laboratoire Suisse d'Analyse du Dopage, Institut Universitaire de Medecine Legale, Rue du Bugnon 21, 1005 Lausanne, Switzerland
Anal Chem (1 June 2005) 77(11): 3579
[PubMed] [Related Article by this group] [Authors contact]

Abstract: "Synthetic insulins such as Humalog Lispro, Novolog Aspart, or Lantus Glargine, are commonly employed for the treatment of insulin-dependent diabetes mellitus owing to convenient handling and fast or prolonged bioavailability. However, the misuse of insulin in sports has been reported often, and the international doping control system requires a reliable and robust assay to determine the presence or absence of related drugs prohibited by the World Anti-Doping Agency. Qualitative evidence of administered substances, which is preferably obtained by mass spectrometry, is of utmost importance. Plasma specimens of 2 mL were fortified with three synthetic insulin analogues and purified by immunoaffinity chromatography, and extracts were analyzed by microbore liquid chromatography and tandem mass spectrometry. Product ion scan experiments of intact proteins enabled the differentiation between endogenously produced insulin and its synthetic analogues by collisionally activated dissociation of multiply charged precursor ions. This top-down sequencing-based assay allows the assignment of individual fragment ions, in particular, of those comprising modifications that are originating from C-termini of B-chains. Recoveries of synthetic insulins from plasma aliquots ranged from 91 to 98%, and detection limits were accomplished at 0.5 ng/mL for all target analytes."

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Proportionate mortality of Italian soccer players: is amyotrophic lateral sclerosis an occupational disease?
Belli MS, Vanacore N
Istituto Superiore di Sanita, Department of Environment and Primary Prevention, Rome, Italy
Eur J Epidemiol (1 January 2005) 20(3): 237
[PubMed] [Authors contact]

Abstract: "OBJECTIVES: The objective of the study is to investigate the mortality experience of Italian soccer players and to discuss the findings in the light of possible long term effects of doping. METHODS: Standardized proportionate mortality ratio (SPMR) and standardized proportionate cancer mortality ratio (SPCMR) were computed for 350 deceased subjects deriving from a list of about 24,000 active Italian soccer players from 1960 to 1996 in the three top leagues (A, B and C). RESULTS: When considering SPMRs, there is a substantial adherence of observed to expected mortality, with the only exception of mortality for diseases of the nervous system (13 obs. vs. 6 exp.) mainly explained by an excess of amyotrophic lateral sclerosis (8 obs. vs 0.69 exp.). As far as SPCMRs are concerned, some digestive cancers (namely: colon cancer, liver cancer and pancreas cancer) show a doubled risk. CONCLUSIONS: A high risk for amyotrophic lateral sclerosis is observed among Italian soccer players. Epidemiological data on association between sport and Amyotrophic Lateral Sclerosis (ALS) are contrasting. On the basis of the overall available evidence we suggest a possible connection between dietary supplements or drugs used to enhance sporting performance and ALS pathogenesis. Further epidemiological studies are needed to confirm these specific mortality risks among soccer players."

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Quantification and profiling of 19-norandrosterone and 19-noretiocholanolone in human urine after consumption of a nutritional supplement and norsteroids
Tseng YL, Kuo FH, Sun KH
Institute of Pharmacology and Toxicology, and Doping Control Center, Tzu Chi University, Hualien, Taiwan
J Anal Toxicol (1 March 2005) 29(2): 124
[PubMed] [Authors contact]

Abstract: "Nandrolone is one of the synthetic anabolic steroids banned in sports and has been a popular substance abused by athletes in recent years. One of its major metabolites, 19-norandrosterone (19-NA), has been used as a determinant for drug violations in sports. Current reports regarding nandrolone-positive cases have been related to intake of some nandrolone-free nutritional supplements. The aim of this study was to learn whether if a nutritional supplement sold by over-the-counter (OTC) nutritional stores could yield the same metabolic products as that of nandrolone. If so, what is (are) the substance(s) that contributed to the nandrolone metabolites? To determine the content of an OTC nutritional supplement, a tablet was dissolved in methanol, followed by N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA)-trimethyliodosilane (TMIS) derivatization prior to gas chromatography-mass spectrometry (GC-MS) analysis. The collected urine samples underwent extraction, enzymatic hydrolysis, and derivatization before the analyses of GC-MS. The results showed that seven anabolic steroids were found as contaminants in the nutritional supplement, in addition to six that were listed in the ingredients by the manufacturer. We confirmed previous reports that administration of the OTC supplement could produce a positive urine test for nandrolone metabolites. Furthermore, the results from excretion studies showed that 19-NA and 19-noretiocholanolone (19-NE) were present in urine after consuming the nutritional supplement, nandrolone, 19-nor-4-androsten-3,17-dione, 19-nor-4-androsten-3beta,17beta-diol, and 19-nor-5-androsten-3beta,17beta-diol. The 19-NA concentrations in urine were generally higher than that of 19-NE (19-NA/19-NE ratio > 1.0) especially during the early stage of excretion, that is, before 6 h post-administration. After this period of time, the concentrations of 19-NA and 19-NE fluctuated and might even have reversed (19-NA/19-NE ratio < 1.0) in their ratio, that is, higher yield in 19-NE than that in 19-NA. On the basis of this study, we postulate that some doping violations of nandrolone could be attributed by indiscriminate administration of the OTC nutritional supplements that contained 19-norsteroids."

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Short-Term administration of supraphysiologic rhGH does not increase maximum endurance exercise capacity in healthy, active young men and women with normal GH-IGF-1 axes
Berggren A, Ehrnborg C, Rosen T, Ellegard L, Bengtsson B-A, Caidahl K
Departments of Clinical Physiology, Endocrinologyand Clinical Nutrition, Sahlgrenska University Hospital, Göteborg; Karolinska Institute, Stockholm, Sweden
J Clin Endocrinol Metab (22 March 2005) doi: 10.1210/jc.2004-1209
[PubMed] [Abstract] [Authors contact]

Abstract: "Context. In spite of the fact that the use of growth hormone (GH) as a doping agent in sports is widespread, little is known about its short-term effects. Objective. To study the effects of GH on exercise capacity. Design. Double-blind, placebo-controlled study; treatment period 28 days. Setting. Subjects from general community studied ambulatory at a university hospital. Participants. 30 healthy active young normal volunteers, 15 female and 15 male, were recruited by local announcement and all completed the study. Intervention. All the subjects were randomized to receive a low GH dose (0.033 mg, or 0.1 IU, per kg/day), a high GH dose (0.067 mg, or 0.2 IU, per kg/day) or placebo. Main outcome measures. Power output and oxygen uptake (VO2) on bicycle exercise. Results. We found no effect by the low or high dosages of GH on maximum oxygen uptake during exercise (mean ± SE for placebo 45.2 ± 1.6 to 45.2 ± 2.1ml/kg/min, GH low dose 42.8 ± 1.6 to 42.8 ± 1.6 ml/kg/min and GH high dose 44.8 ± 3.4 to 44.8 ± 2.2 ml/kg/min; two-way ANOVA not significant). Neither was there any effect on maximum achieved power output during exercise, nor on blood pressure, heart rate or the electrocardiographic ST-level at rest or during exercise. GH significantly increased total body weight (P = 0.028), an effect predominantly ascribed to fluid retention (increased extracellular water volume, ECW), while muscle mass (as indicated by intracellular water volume, ICW) did not change. However, changes of the latter correlated to changes in physical performance, possibly due to different training efforts. Conclusion. Supraphysiologic rhGH during a period of 4 weeks does not improve power output or oxygen uptake. Key words: Doping, exercise capacity, growth hormone, intracellular water, oxygen uptake"

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Seasonal allergy and seasonal decrements in athletic performance
Komarow HD, Postolache TT
Clin Sports Med (1 April 2005) 24(2): e35
[PubMed] [Authors contact]

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Haematological analysis conducted at the departure of the Tour de France 2001
Robinson N, Schattenberg L, Zorzoli M, Mangin P, Saugy M
Int J Sports Med (1 April 2005) 26(3): 200
[PubMed] [Authors contact]

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Modafinil in sports: ethical considerations
Kaufman KR, Gerner R
Br J Sports Med (1 April 2005) 39(4): 241
[PubMed] [Authors contact]

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[Doping control in sports]
Ueki M
Nippon Rinsho (1 December 2005) 62(Supplement 12): 356
[PubMed] [Authors contact]

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[The fight against doping: today and tomorrow]
Rieu M
Bull Acad Natl Med (1 January 2004) 188(6): 955
[PubMed] [Authors contact]

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Development of Miniaturized Competitive Immunoassays on a Protein Chip as a Screening Tool for Drugs
Du H, Wu M, Yang W, Yuan G, Sun Y, Lu Y, Zhao S, Du Q, Wang J, Yang S, Pan M, Lu Y, Wang S, Jing Cheng J
Clin Chem (29 January 2005) 51(2): 368-375
[Abstract] [Authors contact]

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Use of ion trap gas chromatography-multiple mass spectrometry for the detection and confirmation of 3'hydroxystanozolol at trace levels in urine for doping control
Mateus-Avois L, Mangin P, Saugy M
Laboratoire Suisse d'Analyse du Dopage, Institut Universitaire de Medecine Legale, Rue du Bugnon 21, 1005 Lausanne, Switzerland
J Chromatogr B Analyt Technol Biomed Life Sci (25 February 2005) 816(1-2): 193-201 doi:10.1016/j.jchromb.2004.11.033
[PubMed] [Authors contact]

Abstract: "Stanozolol, a synthetic anabolic androgenic steroid, is often abused in sports to enhance performance. Consequently, the anti-doping laboratories daily screen for its metabolites (3'hydroxystanozolol and 4beta hydroxystanozolol) in all urines, mainly by GC-MS, after enzymatic hydrolysis and TMS derivatization. A sensitive and specific method by GC-MS(3) has been developed for the identification in urine of 3'hydroxystanozolol at trace levels. Full mass spectra and diagnostic ions are presented and a case report is commented. In this case, it was possible to attest the presence of a low concentration of stanozolol metabolite in a sample obtained from a competition test. This would have not been possible with other analytical techniques used in the laboratory. Through this case report, it was also possible to demonstrate the importance of sampling and the difficulties that has to face the laboratory when the pre-analytical step is not correctly performed."

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Another designer steroid: discovery, synthesis, and detection of 'madol' in urine
Sekera MH, Ahrens BD, Chang YC, Starcevic B, Georgakopoulos C, Catlin DH
UCLA Olympic Analytical Laboratory, Department of Molecular and Medical Pharmacology, University of California at Los Angeles, 2122 Granville Ave., Los Angeles, CA 90025, USA
Rapid Commun Mass Spectrom (11 February 2005) 19(6): 781-784 doi: 10.1002/rcm.1858
[PubMed] [Authors contact]

Funded by: United States Anti-Doping Agency

Abstract: "Madol (17alpha-methyl-5alpha-androst-2-en-17beta-ol) was identified in an oily product received by our laboratory in the context of our investigations of designer steroids. The product allegedly contained an anabolic steroid not screened for in routine sport doping control urine tests. Madol was synthesized by Grignard methylation of 5alpha-androst-2-en-17-one and characterized by mass spectrometry and NMR spectroscopy. We developed a method for rapid screening of urine samples by gas chromatography/mass spectrometry (GC/MS) of trimethylsilylated madol (monitoring m/z 143, 270, and 345). A baboon administration study showed that madol and a metabolite are excreted in urine. In vitro incubation with human liver microsomes yielded the same metabolite. Madol is only the third steroid never commercially marketed to be found in the context of performance-enhancing drugs in sports."

Article intro: "Rumors of designer steroids, conceived and used specifically to evade sport drug testing, have been festering for several years. In 2002 we found norbolethone, a never-marketed steroid, in two urine samples and speculated that it was recently synthesized as a steroid that would not be detected because it was not named on the prohibited list.[1] After we discovered tetrahydrogestrinone (THG) in a syringe provided to us by the United States Anti-Doping Agency in 2003,[2] there was no doubt that a clandestine designer steroid industry existed. Recently, an oily product allegedly containing an anabolic androgenic steroid not screened for in routine sport doping control urine tests was received by our laboratory. We describe herein the identification of the unknown compound contained in the oily product, its synthesis, a baboon excretion study, a human liver microsome incubation experiment, and a gas chromatography/mass spectrometry (GC/MS) method for detection and identification in urine. The method was applied at the UCLA Olympic Laboratory and at the 2004 Olympic Games in Athens..."

Note added in proof: "On February 1, 2005, the World Anti-Doping Agency (WADA) announced the independent discovery by scientists at INRS Sante, Montreal, of a designer steroid which the scientists named desoxymethyltestosterone (DMT). Based on a communication from INRS Sante, DMT is 17-methyl-5-androst-2-en-17-ol. This is madol."

AK note: Baboon madol excretion study has apparent n=1 and thus is not statistically valid.

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Characterization of chemically modified steroids for doping control purposes by electrospray ionization tandem mass spectrometry
Thevis M, Bommerich U, Opfermann G, Schanzer W
Institute of Biochemistry, German Sport University Cologne, Carl-Diem Weg 6, 50933 Cologne, Germany
J Mass Spectrom (15 February 2005)
[PubMed] [Authors contact]

Abstract: "The discovery of the designer steroid tetrahydrogestrinone (THG) in elite athletes' doping control samples in 2003 demonstrated the availability of steroid derivatives prepared solely for doping purposes. Modern mass spectrometers utilizing electrospray ionization and collisionally activated dissociation (CAD) of analytes allow the structural characterization of steroids and their derivatization sites by the elucidation of fragmentation behaviors. A total of 21 steroids comprising either a 4,9,11-triene, a 3-keto-4-ene or a 3-keto-1-ene nucleus were investigated regarding their dissociation pathways, deuterated analogues were synthesized and fragmentation routes were postulated, permitting the identification of steroidal structures and modifications. Compounds based on a 4,9,11-triene steroid with an ethyl residue at C-13 (gestrinone analogues) generate abundant fragment ions at m/z 241 and 199, whereas the substitution of the C-13 ethyl group by a methyl residue (trenbolone analogues) results in a shift of m/z 241 to 227. Substances related to testosterone with a 3-keto-4-ene structure give rise to abundant fragment ions at m/z 109 and 97 whereas steroids with a 3-keto-1-ene nucleus eliminate the A-ring including the carbons C-1-C-4, in addition to C-19 that is proposed to migrate from C-10 to C-1 under CAD conditions."

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Postoperative course and anabolic-androgenic steroid abuse -- a case report
Medras M, Tworowska U, Jozkow P, Dumanski A, Dubinski A
Department of Sports Medicine, University of Physical Education, Wroclaw, Poland
Anaesthesia (January 2005) 60(1): 81-84
[PubMed] [Authors contact]

Abstract: "It is estimated that 80% of weight lifters and body-builders take anabolic-androgenic steroids. Their long-term use is associated with a variety of pathological conditions and premature death. Anabolic-androgenic steroid abuse may lead to changes in the presentation and progression of some conditions. It remains unclear whether anabolic steroids should be given to patients with a history of abuse of these drugs who are to undergo surgery. We report on a fatal outcome following surgery in a 48-year-old weight lifter."

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Detection of physiological concentrations of cortisol and cortisone in human hair
Raul JS, Cirimele V, Ludes B, Kintz P
Clin Biochem (December 2004) 37(12): 1105-11
[PubMed] [Authors contact]

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World Medical Association Declaration on Principles of Health Care for Sports Medicine
World Medical Association
Bull Med Ethics (December 1999) (153): 10-11
[PubMed] [Authors contact]

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Drugs, sport and the Olympics 2000-2004
Millar AP
Director of Research, Lewisham Sports Medicine Clinic, 1 West Street, Petersham, NSW 2049
Med J Aust (15 November 2004) 181(10): 584
[PubMed] [Authors contact]

Excerpt: "There is a ban on oxygen-transport drugs and on physical environment enhancers such as hypobaric chambers. Both are alleged to produce the same result, but only use of the drug can be tested. The penalty for the drug user is disqualification, but for the hypobaric enthusiast a rousing cheer for a drug-free effort. The crime is the same, so why vary the penalty? There is never likely to be a level playing field under the present system, in which one reads of positive test results being swept under the table. How will drug testing eliminate the genetic inequalities between athletes? How will testing improve the availability of top-level coaches and training facilities to all? How can it eliminate the inequality in financial incentives, allowing some athletes to train for 6 hours daily while others have to work to enable them to train for even 2 hours daily? We have swimming costumes that decrease drag in the water, resulting in faster times. These are not universally available, giving their owners an advantage. A level playing field will never exist in our present system. It is incongruous that in all this mess, only drugs are available to all. The current frenzy to test blood has ethical problems which have not been addressed. What is to happen to an athlete who develops an infection from a dirty needle? Who is responsible for the tester who has a needlestick injury from an HIV-positive athlete? It is worth remembering that this diagnosis will only be made 3 months after the Games, when everyone has dispersed. The whole area needs to be reviewed by an outside body with no vested interest in the outcome."

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WADA’s 2005 List of Prohibited Substances and Methods
WADA
WADA Web Resources (effective January 2005)
[FullText (.PDF)]

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Longitudinal variation of hemoglobin and reticulocytes in elite rowers
Ashenden MJ, Lacoste A, Orhant E, Audran M, Sharpe K.
Haematologica (November 2004) 89(11): 1403-1404
[PubMed] [Authors contact]

Abstract: "Longitudinal monitoring of athlete's hematologic parameters holds considerable promise as a strategy to detect and thereby deter illicit blood doping. This study documents temporal changes of hemoglobin concentration (Hb) and reticulocyte counts in elite rowers. The 'within subject' variation in rowers was comparable to that of athletes from other sports. Reticulocyte results were dependent on the type of instrument used."

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"Genetic Doping" with erythropoietin cDNA in primate muscle is detectable
Lasne F, Martin L, de Ceaurriz J, Larcher T, Moullier P, Chenuaud P.
Laboratoire National de Dépistage du Dopage, 143 Avenue Roger Salengro, 92290, Châtenay-Malabry, France.
Mol Ther (September 2004) 10(3): 409-410
[PubMed] [Authors contact]

Leading Text: "Forthcoming “genetic doping” is predicted to be undetectable. In the case of recombinant human erythropoietin (rhEPO), a hormone used in endurance sports, it is being predicted that exogenous drug injections will be replaced by the transfer of the corresponding gene into some of the athlete’s own cells. The hormone thus produced inside the organism is assumed to be completely identical to the physiological one. Our results show that this is not the case and open up optimistic prospects for antidoping control involving gene transfer.

Doping in sport, with very few exceptions, arises from misused medical treatments. This is the case for rhEPO, a hormone that stimulates red blood cell production and that has become a key element of doping in endurance sports. Treatment with rhEPO currently requires repeated injections of recombinant hormones obtained from nonhuman cells, i.e., Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, into which the human gene of the hormone has been inserted. Natural endogenous and rhEPO were shown to present different isoelectric profiles, probably the result of altered posttranslational modifications that are species- and tissue type-dependent. This difference has allowed for the development of a test to detect the presence of rhEPO in urine, a test that is currently used in antidoping controls [1].

Genetic technologies are expected to change the very nature of medical treatments. For instance, it is now conceivable that administration of an exogenous therapeutic protein will be replaced by introducing the corresponding gene into some of the patient’s own cells. It is almost inevitable that athletes will exploit such medical progress in an effort to elude detection by sport authorities charged with curbing doping practices. Doping practices, in addition to being the focus of regulatory issues, may also severally and adversely affect the health of athletes that engage in such practices. Doping by gene transfer may compound these adverse side effects because of direct toxic effects, persistent gene expression, or potential insertional mutagenesis [2] and [3]. Furthermore, the assumption that these new methods of doping will yield proteins that are identical to the endogenous gene product, thus making detection impossible, may not be the case.

To compare the isoelectric profiles of physiological EPO and hormone resulting from in vivo gene transfer, we have adapted for serum analysis a method previously developed for urine [4]. Using this method, samples from cynomolgus macaques were analyzed for the serum recombinant EPO profile before and after transfer of the homologous cDNA into skeletal muscle by injection of recombinant adeno-associated virus [5]. Transgene expression was controlled by a doxycycline-regulatable system [6]..."

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Detection of isoelectric profiles of erythropoietin in urine: differentiation of natural and administered recombinant hormones
Lasne F, Martin L, Crepin N, de Ceaurriz J.
Laboratoire National de Dépistage du Dopage, 143 Avenue Roger Salengro, 92290, Châtenay-Malabry, France.
Anal Biochem (15 December 2002) ePub 12 November 2002 10(3): 409-410
[PubMed] [Authors contact]

Abstract: "Erythropoietin (EPO) is normally present in urine at a low concentration (about 1IU/L, i.e., about 10ng/L) for a total protein concentration of at least 50mg/L. A method to study the isoelectric profile of this hormone from 20-ml urine aliquots without previous purification was developed. This method involves isoelectric focusing of the retentate from ultrafiltered urine. Both the ultrafiltration and the isoelectric focusing required precautionary measures to prevent EPO degradation by the proteases that are present in urine. Because classical immunoblotting gave rise to an unspecific detection of various urinary proteins in the focused retentate, it was essential to use the "double-blotting" process developed to solve this problem. Sufficient sensitivity was achieved using amplified chemiluminiscent detection after the blotting membrane was treated with dithiotreitol. The patterns that were revealed from various urinary samples proved to be highly heterogeneous as they were composed of more than 10 isoforms in a pI range of 3.7-4.7. Clear transformation of the patterns was observed in the case of treatment by the recombinant hormone, suggesting that this method can be regarded an efficient tool for indicating recombinant EPO misuse in sports. It may also open new investigations in the field of physiologic or pathologic exploration."

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