WADA ... Olympics ...

Tort N, Salvador JP, Marco MP. Multiplexed immunoassay to detect anabolic androgenic steroids in human serum. Anal Bioanal Chem 2012;403(5):1361-71. http://www.springerlink.com/content/q94w723050111536/ (Analytical and Bioanalytical Chemistry, Volume 403, Number 5 - SpringerLink)

A multianalyte enzyme-linked immunosorbent assay (ELISA) has been developed for the simultaneous detection of anabolic androgenic steroids (AAS) in human serum. The multiplexed method was developed according to a planar strategy in which the analytes are identified by their location in the microtiter plate. In the immunochemical procedure established here, human serum samples are mixed with a cocktail of antibodies and added to the distinct sections of a microplate biofunctionalized with different haptenized biomolecules. The cocktail of antibodies consists of a mixture of polyclonal antibodies raised against stanozolol (ST), boldenone (B), and tetrahydrogestrinone (THG). The whole immunochemical analytical procedure takes around 2 h including sample preparation, and many samples can be processed simultaneously to screen for the presence of the three AAS in a single run. Using this ELISA, ST, B, and THG can be detected and quantified individually. When used as a screening method, due to the cross-reactivity profiles of the immunoreagents used, the presence of up to 11 AAS can be detected simultaneously. The detectabilities achieved by this method in human serum are below the MRPLs (minimum required performance limits) proposed by WADA (World Anti-Doping Agency) and reference laboratories of the European Community.

all very interesting. i hope this translates into other areas.
but biological passport is a hormone panel, i think, taken over time and any changes noted. then non analytical rules come into play. this looks like a new high tech drug test. i may be wrong.
it says up to 11 can be detected due to cross reactivity without naming them. any ideas?
why those three. stanozolol a derivative of dht. boldenone of testosterone. thg a progestin. i think. maybe this?
 
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First doping violation concluded in Athletics on the basis of the Athlete Biological Passport

Monaco – Hélder Ornelas, 38, a long distance runner from Portugal, has been found guilty of a doping violation and sanctioned with a 4-year period of ineligibility by the Portuguese Athletic Federation (FPA).

The evidence against Ornelas comprised of a series of blood test results collected by the IAAF in the course of the Athlete Biological Passport programme between December 2009 and November 2010. It is the first time that the Athlete Biological Passport has been used in Athletics as sole evidence in support of an anti-doping rule violation.

The Athlete Biological Passport measures and monitors an athlete’s blood variables over time and establishes an individual longitudinal profile which can indicate the use of a prohibited substance or a prohibited method.

Ornelas’s blood profile was flagged as being abnormal in May 2011 which triggered further investigations in accordance with IAAF Anti-Doping Regulations. Ornelas’s blood profile was submitted to an Expert Panel composed of three international experts in the field of haematology. Following an in-depth review of Ornelas’s profile, the experts unanimously concluded that there was no known reasonable explanation for the abnormalities observed in his blood profile other than the use of a prohibited substance or a prohibited method.

On the basis of the experts’ conclusion, the IAAF initiated disciplinary proceedings against Ornelas and referred the case to the Portuguese Athletic Federation for adjudication. In referring the case to the FPA, the IAAF sought a 4-year sanction as it considered that there were aggravating circumstances in this case justifying the imposition of an increased sanction. This is in line with the IAAF’s long-standing position in favour of an increased sanction for serious first time doping offences and it will continue to enforce the rules on aggravating circumstances whenever there are valid grounds for doing so.

The FPA decision concludes a 10-month evaluation and adjudication process. Ornelas did not exercise his right of appeal to the Court of Arbitration for Sport and the FPA’s decision has become final and binding under IAAF Rules.

IAAF President Lamine Diack said: "Those who try to cheat within the athletics community should be warned that the Athlete Biological Passport is not merely a concept but rather an efficient method that is now being used by the IAAF Anti-Doping Department to identify, target and catch those who believe that doping is the only route to success. Cheaters should also be aware that, if they are caught, the IAAF will seek an increased 4-year sanction whenever the circumstances so justify."
 
Bosch J, Ueki M, Such-Sanmartin G, Segura J, Gutierrez-Gallego R. Tracking growth hormone abuse in sport: A comparison of distinct isoform-based assays. Anal Chim Acta 2012;733:56-63. ScienceDirect.com - Analytica Chimica Acta - Tracking growth hormone abuse in sport: A comparison of distinct isoform-based assays

Detecting recombinant human growth hormone (rhGH) abuse in sport remains one of the major challenges in doping control. We have compared two different approaches to detect the hGH (human growth hormone) abuse. The first measures the concentrations of the 22kDa hGH isoform (rec assay) and pituitary derived isoforms (pit assay) and a ratio rec/pit is obtained. The second measures the concentrations of 22 and 20kDa hGH isoforms and also a ratio 22/20kDa is derived. Using a single set (nine healthy male subjects, 7 days, 0.026mg/kg/day of rhGH, 2 week wash out period) both approaches were compared. To quantify the agreement between the immunoassays, B.A. (Bland-Altman) analysis and P.r. (Pearson correlation) were used. To fully understand the assay readings, all relevant antibodies were characterised by surface plasmon resonance (SPR). In either approach the ratio numerator produces similar results and the denominator determines both signal-amplitude and time-frame of possible application. The rec vs pit approach displays a higher distinctive capacity to detect hGH abuse but the complex binding properties of the capture antibodies make it very difficult to evaluate the precise contributions of the individual hGH variants to the assay result. In the 22 vs 20 approach, the 20kDa hGH concentration measures determine its applicability. Both approaches are based on a different principle, should be preferably applied within 24h after rhGH administration, and are perfectly comparable given the results obtained. The reduced time frame of application indicates that their principle application should be preferably in an out-of-competition setting.
 
Baumann GP. Growth Hormone Doping in Sports: A Critical Review of Use and Detection Strategies. Endocrine Reviews 2012;33(2):155-86. http://edrv.endojournals.org/content/33/2/155.abstract

GH is believed to be widely employed in sports as a performance-enhancing substance. Its use in athletic competition is banned by the World Anti-Doping Agency, and athletes are required to submit to testing for GH exposure. Detection of GH doping is challenging for several reasons including identity/similarity of exogenous to endogenous GH, short half-life, complex and fluctuating secretory dynamics of GH, and a very low urinary excretion rate. The detection test currently in use (GH isoform test) exploits the difference between recombinant GH (pure 22K-GH) and the heterogeneous nature of endogenous GH (several isoforms). Its main limitation is the short window of opportunity for detection (∼12–24 h after the last GH dose). A second test to be implemented soon (the biomarker test) is based on stimulation of IGF-I and collagen III synthesis by GH. It has a longer window of opportunity (1–2 wk) but is less specific and presents a variety of technical challenges. GH doping in a larger sense also includes doping with GH secretagogues and IGF-I and its analogs. The scientific evidence for the ergogenicity of GH is weak, a fact that is not widely appreciated in athletic circles or by the general public. Also insufficiently appreciated is the risk of serious health consequences associated with high-dose, prolonged GH use. This review discusses the GH biology relevant to GH doping; the virtues and limitations of detection tests in blood, urine, and saliva; secretagogue efficacy; IGF-I doping; and information about the effectiveness of GH as a performance-enhancing agent.
 
Bosch J, Ueki M, Such-Sanmartin G, Segura J, Gutierrez-Gallego R. Tracking growth hormone abuse in sport: A comparison of distinct isoform-based assays. Anal Chim Acta 2012;733:56-63. ScienceDirect.com - Analytica Chimica Acta - Tracking growth hormone abuse in sport: A comparison of distinct isoform-based assays

Detecting recombinant human growth hormone (rhGH) abuse in sport remains one of the major challenges in doping control. We have compared two different approaches to detect the hGH (human growth hormone) abuse. The first measures the concentrations of the 22kDa hGH isoform (rec assay) and pituitary derived isoforms (pit assay) and a ratio rec/pit is obtained. The second measures the concentrations of 22 and 20kDa hGH isoforms and also a ratio 22/20kDa is derived. Using a single set (nine healthy male subjects, 7 days, 0.026mg/kg/day of rhGH, 2 week wash out period) both approaches were compared. To quantify the agreement between the immunoassays, B.A. (Bland-Altman) analysis and P.r. (Pearson correlation) were used. To fully understand the assay readings, all relevant antibodies were characterised by surface plasmon resonance (SPR). In either approach the ratio numerator produces similar results and the denominator determines both signal-amplitude and time-frame of possible application. The rec vs pit approach displays a higher distinctive capacity to detect hGH abuse but the complex binding properties of the capture antibodies make it very difficult to evaluate the precise contributions of the individual hGH variants to the assay result. In the 22 vs 20 approach, the 20kDa hGH concentration measures determine its applicability. Both approaches are based on a different principle, should be preferably applied within 24h after rhGH administration, and are perfectly comparable given the results obtained. The reduced time frame of application indicates that their principle application should be preferably in an out-of-competition setting.
 
Faster, stronger – and high
The development of genetic enhancement in animals has led to fears that ‘gene doping’ could be abused by athletes
Faster, stronger – and high - FT.com

August 3, 2012 9:24 pm
By Clive Cookson

Meet Marathon Mouse. The rodent genetically engineered in the US can run 5km nonstop on a treadmill, compared with an average of 0.2km in its wild cousins.

The emergence of animals such as Marathon Mouse has led to fears that genetic enhancement will soon boost human sporting performance. Indeed rumours have been circulating for several years of athletes succumbing to the temptation of “gene doping”.

Meanwhile, anonymous accusers maintain that doping with more conventional drugs such as steroids still pervades elite sport, despite the huge clean-up effort made by the World Anti-Doping Agency and the governing bodies of individual sports.

Such suspicions gained fresh momentum at the Olympics this week with the performance of China’s Ye Shiwen. The 16-year-old swimming prodigy completed the last 50m of her 400m medley faster than the men’s champion, leading an American coach pointedly to brand her performance “unbelievable”. Yet she has passed four official drug tests in the past 12 months, including this week at the games.

Chatter in the sporting world about widespread cheating – for example, by athletes taking finely calibrated quantities of drugs during tightly controlled periods, perhaps with the addition of masking agents – contrasts with the view of officials from testing authorities that they are gaining ground in the never-ending race to keep ahead of the cheats.

“Olympic sports are cleaner in the London games than ever before,” says Professor Tony Moffat of London University’s School of Pharmacy, who has more than 40 years experience of sports doping analysis. “The new anti-doping lab is the most sophisticated we have seen. They will detect any banned substance.”

The £20m laboratory set up for the 2012 games in GlaxoSmithKline’s research centre about 30 miles north of London will be analysing 1,200 blood and 3,800 urine samples during the Olympics. One thousand staff, including 150 scientists, are working round the clock to test about half the competitors, chosen at random, for 240 banned substances. In addition, all medal winners are tested.

By Friday, after the first week of competition, at least three athletes had been kicked out for doping offences. They include Albanian weightlifter Hysen Pulaku tested positive for stanozolol, the anabolic steroid that achieved notoriety in 1988, when Canadian sprinter Ben Johnson was stripped of his gold medal after failing a test for the same drug. And Uzbek gymnast Luiza Galiulina was banned for using furosemide, a diuretic that can help with weight loss and act as a masking agent making it harder to detect other drugs.

But, with a typical lag of three to six days between a sample being taken and naming and shaming,it is not unlikely that cheats will be revealed in greater numbers next week and beyond. In Beijing 2008, a total of 14 athletes failed drugs tests.

“Even if athletes believe they are taking a banned substance that cannot be detected, they should remember that their samples are frozen and stored for eight years,” says Prof Moffat. “They can be caught through retrospective analysis, if a test becomes available, and any Olympic titles removed.”

As anti-doping procedures grow more sophisticated in step with emerging drug misuse techniques, several tests are being used for the first time in London. The authorities are particularly pleased with improved methods of detecting human growth hormone. Synthetic HGH has recently been favoured by cheats who want to build muscle strength without risking the side-effects of anabolic steroids, including damage to the metabolism. It is also hard to detect since it is chemically almost identical to the hormone produced naturally by the body. Richard Holt of Southampton university, who helped to develop the new test, says it can reliably detect the substance, even a few weeks after the last dose has been administered.

Steroids – once administered routinely by state sports administrators in East Germany and elsewhere in the Soviet bloc – are the classic muscle-building drugs. Today, the authorities are constantly on the lookout for new “designer steroids” created by underground chemists to avoid detection. The big buzz surrounds synthetic selective androgen receptor modulators, which have fewer side effects than conventional steroids. They are banned by Wada but are widely available on black market websites. The authorities, though geared up to detect them, have not yet caught anyone.

Sports doping is a malign spin-off from legitimate medicines designed to treat diseases such as muscle wasting and anaemia. The primary aims of athletes who abuse such products is to build muscle strength and mass, improve energy metabolism and increase the blood’s oxygen-carrying capacity. Doping through blood boosting, for example, started in earnest during the 1990s when the hormone erythropoietin (EPO) became available as a medical treatment. Many athletes, particularly in endurance sports such as cycling, took it to produce more red blood cells, which would carry more oxygen to their muscles.

Given the industry’s unwitting role in the development of sports doping, Wada was delighted last year when the International Federation of Pharmaceutical Manufacturers and the Biotechnology Industry Organization agreed that member companies should pass on confidential data about drugs in clinical trials with potential for abuse in sport. GSK, for example, has told Wada about a pill in its pipeline that boosts red blood cell production.

Wada has been preparing since 2002 for the advent of gene doping – an abuse of gene therapy, a medical procedure in advanced clinical trials to treat a wide range of disorders from cystic fibrosis to blood disease. Athletes would be given extra copies of genes that make performance-enhancing proteins such as growth factors in their muscles. But despite spectacular results achieved with laboratory animals such as Marathon Mouse, which makes an enzyme called PEPCK-C, scientists say formidable difficulties would face anyone trying to transfer the technology to humans.

“Although we know that some gene therapy experts have been approached by athletes or coaches and there are rumours of gene doping being used, there is no firm evidence for this,” says Olivier Rabin, Wada’s chief scientist. According to UK Anti-Doping, such abuse could not at present be reliably detected. However, scientists hope to develop tests for added genes or the viruses used to carry them into human cells.

...

Wada, meanwhile, is placing more emphasis on the “athlete biological passport” – an indirect catch-all method of detection. This establishes a long-term metabolic profile for each individual, with baseline levels of hormones and other protein biomarkers in his or her blood. Any sudden, performance-enhancing change that cannot be explained legitimately is regarded as evidence of doping, even if there is no direct detection of a banned substance.

Though still in their infancy, biological passports are already exposing cheats. Just before the Olympics started, the International Association of Athletics Federations announced that six athletes (three Russians, one Turk, one Greek and one Moroccan) had been banned for either two or four years on the basis of such records.

At the same time, sports scientists are reminding people that improvements in performance, even as outstanding as those of Chinese swimmer Ms Ye, should not in themselves be taken as evidence of cheating. The pool of potential competitors is expanding, particularly in the developing world and in countries where women are participating more actively. And science, technology and investment are improving infrastructure at elite level – nutrition, training facilities and equipment – which is pushing performance and creating new records everywhere.

Indeed, a new report by the UK Institution of Mechanical Engineers warns of the dangers of “technology doping” in the years ahead, and calls on sporting regulators to work with engineers to predict the consequences of new technologies.

“Engineering has had an enormous and under-appreciated influence on sport over the past 100 years,” says lead author Philippa Oldham. “Almost every sport, from athletics to cycling, has benefited from the introduction of new materials, techniques and tools.”

Yet rumours of pharmaceutical and biological cheating will not go away. “Yes there are persistent myths about super-athletes being produced in hollowed-out volcanoes,” jokes Graham Arthur of UK Anti-Doping. “But the overwhelming majority of athletes want to be clean. They value the benefits of competing in a clean sports environment.”

Even so, it is not certain that the technology that led to Marathon Mouse will remain permanently caged in the animal lab. “It is like an arms race,” says Andy Parkinson, chief executive of UK Anti-Doping. “We need to keep pressing and not think that we’ve tackled it [permanently] because I don’t think we ever will.”
 
Riedmaier I, Benes V, Blake J, et al. RNA-Sequencing as Useful Screening Tool in the Combat against the Misuse of Anabolic Agents. Anal Chem. RNA-Sequencing as Useful Screening Tool in the Combat against the Misuse of Anabolic Agents - Analytical Chemistry (ACS Publications)

The abuse of anabolic substances in animal husbandry is forbidden within the EU and well controlled by detecting substance residues in different matrices. The application of newly designed drugs or substance cocktails represents big problems. Therefore developing sensitive test methods is important. The analysis of physiological changes caused by the use of anabolic agents on the molecular level, for example, by quantifying gene expression response, is a new approach to develop such screening methods. A novel technology for holistic gene expression analysis is RNA sequencing. In this study, the potential of this high-throughput method for the identification of biomarkers was evaluated. The effect of trenbolone acetate plus estradiol on gene expression in liver from Nguni heifers was analyzed with RNA sequencing. The expression of 40 selected candidate genes was verified via RT-qPCR, whereby 20 of these genes were significantly regulated. To extract the intended information from these regulated genes, biostatistical tools for pattern recognition were applied and resulted in a clear separation of the treatment groups. Those candidate genes could be verified in boars and in calves treated with anabolic substances. These results show the potential of RNA sequencing to screen for biomarker candidates to detect the abuse of anabolics. The verification of these biomarkers in boars and calves leads to the assumption that gene expression biomarkers are independent of breed or even species and that biomarkers, identified in farm animals could also act as potential biomarker candidates to detect the abuse of anabolic substances in human sports.
 
Bosch J, Such-Sanmartin G, Segura J, Gutierrez-Gallego R. Tracking growth hormone abuse in sport: Performance of marker proteins in a controlled setting. Anal Chim Acta 2012;745C:118-23. ScienceDirect.com - Analytica Chimica Acta - Tracking growth hormone abuse in sport: Performance of marker proteins in a controlled setting

Human growth hormone (hGH) abuse in sport is a challenge at present. The current strategy used, known as direct method, is based on the quantification of hGH variants in serum. An alternative strategy, known as indirect method, focuses on serum markers such as insulin-like growth factor I (IGF-I) and procollagen type III N-terminal propeptide (P-III-NP). The indirect method allows a longer window of detection (WOO) of hGH abuse. To evaluate the performance of the indirect method, in parallel to the direct method, a clinical trial with recombinant hGH (rhGH) was conducted on healthy male subjects during 7 days (0.026mg(-1)kg(-1)person(-1)day(-1)). The data were fit to the discriminant formula proposed in the previously published GH-2000 project. The low sensitivity of the scores, judged from the high number of false negative outcomes, imposed a new discriminant analysis, standarised using local population subjects demographically similar to the ones of the study. The sensitivity of the method significantly increased, highlighting the importance of the standardisation. The indirect method allowed extended window of opportunity (WOO), although two false positive evaluations were observed derived from elevated basal IGF-I and P-III-NP concentrations stressing the need for an independent confirmation method. When direct and indirect methods were combined the best selectivity and sensitivity were achieved.
 
Jing J, Zhou X, He C, et al. Biomarker detection of rhGH Doping: an excretion study. Drug Testing and Analysis. Biomarker detection of rhGH Doping: an excretion study - Jing - 2012 - Drug Testing and Analysis - Wiley Online Library

The purpose of this research is to validate the biomarker-based approach for the detection of doping with recombinant human growth hormone (rhGH) in sport. The GH-2000 project proposed an indirect method for the detection of exogenously administered growth hormone (GH) based on the measurement of the GH-dependent markers: insulin-like growth factor-I (IGF-I) and Type III pro-collagen (P-III-P). These markers rise in a dose-dependent manner after GH application. In this study, the concentrations of IGF-I, IGF-BP3, and P-III-P in serum were determined to provide further incentives for the implementation of this detection assay in modern anti-doping programmes. This paper reports on an administration study of rhGH involving 25 Chinese male volunteers at a dose of 0.1?IU /kg/day for a continuous 14-day period.

We observed that the serum IGF-I concentration increased rapidly in the rhGH treatment group and showed significantly higher levels compared to baseline between days 4 and day 16 after administration. Although the response of P-III-P to rhGH administration was delayed compared to the IGF-I axis, the P-III-P concentration remained increased for a longer period (from day 4 to day 28). Statistical analysis was carried out to establish a discriminant formula with Statistical Product and Service Solutions (SPSS) concluding that the biomarker methodology is valid and universally applicable.
 
Detection Of Designer Steroid Methylstenbolone In "Nutritional Supplement"

Highlights
? Detection of designer steroid methylstenbolone in “nutritional supplement”.
? Identification of two long-term 16-hydroxylated methylstenbolone metabolites.
? Good targets to monitor methylstenbolone misuse in doping control analysis.
? Elucidation of metabolites structures using gas chromatography and mass spectrometric.


Cavalcanti GD, Leal FD, Garrido BC, Padilha MC, Neto FR. Detection of designer steroid methylstenbolone in "nutritional supplement" using gas chromatography and tandem mass spectrometry: elucidation of its urinary metabolites. Steroids. ScienceDirect.com - Steroids - Detection of designer steroid methylstenbolone in “nutritional supplement” using gas chromatography and tandem mass spectrometry: Elucidation of its urinary metabolites

The use of "nutritional supplements" containing unapproved substances has become a regular practice in amateur and professional athletes. This represents a dangerous habit for their health once no data about toxicological or pharmacological effects of these supplements are available. Most of them are freely commercialized online and any person can buy them without a medical surveillance. Usually, the steroids intentionally added to the "nutritional supplements" are testosterone analogs with some structural modifications. In this study, the analyzed product was bought online and a new anabolic steroid known as methylstenbolone (2,17alpha-dimethyl-17beta-hydroxy-5alpha-androst-1-en-3-one) was detected, as described on label. Generally, anabolic steroids are extensively metabolized, thus in-depth knowledge of their metabolism is mandatory for doping control purposes. For this reason, a human excretion study was carried out with four volunteers after a single oral dose to determine the urinary metabolites of the steroid. Urine samples were submitted to enzymatic hydrolysis of glucuconjugated metabolites followed by liquid-liquid extraction and analysis of the trimethylsilyl derivatives by gas chromatography coupled to tandem mass spectrometry. Mass spectrometric data allowed the proposal of two plausible metabolites: 2,17alpha-dimethyl-16xi,17beta-dihydroxy-5alpha-androst-1-en-3-one (S1), 2,17alpha-dimethyl-3alpha,16xi,17beta-trihydroxy-5alpha-androst-1-ene (S2). Their electron impact mass spectra are resonable to that of 16-hydroxylated steroids presenting diagnostic ions such as m/z 231 and m/z 218. These metabolites were detectable after one week post administration while unchanged methylstenbolone was only detectable in a brief period of 45 h.
 
Personally I think the answer, for me anyway, is to just not care about the Olympics.

For each individual sport, I just don't think the Olympics represent the pinnacle.

Except perhaps in sports that no one cares about in the first place, that really have no other major competition.

E.g., we just don't need the Olympics to find who is the fastest sprinter in the world right now, who is the highest-totalling PL'er, or who is the best soccer team, etc.

Is it possible that part of the extreme doping-obsession isn't just the sanctity, sports-worship aspect combined with the complexes resulted from being stuffed in lockers by the jocks in high school, but also that "only the Olympics" will do this extreme testing so it's your one place to go for "pure" sports? In contrast to say powerlifting competitions or sprint meets or soccer games run by other organizations.

As to the researchers, they are of course trying to get money to fund their careers.

When having to go to these obsessive extremes, which seem to be driven more by distorted psychology, attempts to manipulate the public, and personal gain than anything else, my interest becomes lost.

Similarly, for example even if I never used anabolic steroids, HCG, or GH, if some employer wanted to run 50 different "biochemical profile" tests on me that I had to pass, complete with urine, hair, fingernail clipping, skin cell, and who knows perhaps stool samples my answer would be that I will go elsewhere. Not for me. Take your competition and shove it, basically. Or, as the case may be, we can sit out the season, etc.

The MLB players succeeded a long time with that basic approach but unfortunately didn't manage to hold the line :(

And then besides all the above, I want my baseball players, sprinters, etc to be juiced anyway. I don't care to spend my time watching sub-par performances :)

Word..
I agree 100%.. I mean look at Derek Eater / Jeter. He will manage to get a xript for recovery and have 1 of his best years.. imo if these NFL and MLB players are getting caught then its just pure Greed / pushing the envelope.
 
Ekstrom L, Cevenini L, Michelini E, et al. Testosterone challenge and androgen receptor activity in relation to UGT2B17 genotypes. Eur J Clin Invest. Testosterone challenge and androgen receptor activity in relation to UGT2B17 genotypes - Ekstr[]m - 2013 - European Journal of Clinical Investigation - Wiley Online Library

BACKGROUND: We investigated the androgen receptor (AR) bioluminescense response in serum and urine before and after testosterone challenge in different genotypes of the UGT2B17 enzyme, which catalyses testosterone glucuronidation.

MATERIAL AND METHODS: The androgen receptor activity was determined using a yeast-based bioluminescence assay. The androgens were analysed using LC-MS/MS, and the individuals were genotyped for UGT2B17 deletion polymorphism using real-time polymerase chain reaction.

RESULTS: The serum concentrations of testosterone and dihydrotestosterone (DHT) were markedly elevated on days 2 and 4 and were still above baseline on day 15 after a dose of 500 mg testosterone enanthate. The androgenic activity in serum increased in parallel and correlated with the hormone concentrations and remained above baseline on day 15. The urinary androgenic activity increased 4-5-fold and was closely related to the unconjugated testosterone and independent of the UGT2B17 genotype.

CONCLUSIONS: The AR assay may serve as a complement to the urinary testosterone/epitestosterone (T/E) doping test, because this is profoundly influenced by the UGT2B17 deletion polymorphism. It may also be useful for detection of other illicit androgens in sports, or in the society, or for monitoring and diagnostics of androgen-related disorders.
 
Baseball to Expand Drug-Testing Program
http://www.nytimes.com/2013/01/11/s...agree-to-in-season-blood-testing-for-hgh.html

Published: January 10, 2013
By MICHAEL S. SCHMIDT

WASHINGTON — A day after suffering through the embarrassment of seeing no new players selected to the Hall of Fame, Major League Baseball and its players union moved ahead Thursday by announcing that they had reached an agreement to expand their drug-testing program.

For the first time, baseball will conduct in-season blood testing for human growth hormone. In addition, it will employ a new test designed to catch players using synthetic testosterone, a substance that appears to be growing in popularity because it washes out of a player’s body fairly quickly after being used, making it hard to detect.

The expansion of the testing program allows Major League Baseball to again argue that it has moved ahead of the National Football League on the drug front and that it now has the toughest testing program of any of the professional sports leagues in North America. The N.F.L. has yet to test for H.G.H. and does not have a comparable testosterone test. In 2011, the N.F.L. and its players union said they had agreed to initiate blood testing for H.G.H., but since then the union has expressed reservations and no testing protocol has been established.

The disclosure that there would be expanded testing in baseball came 24 hours after the members of the Baseball Writers’ Association of America delivered a tough verdict on the role that performance-enhancing drugs have played in the sport, emphatically rejecting the first-time candidacies of Barry Bonds and Roger Clemens because of their direct links to such substances.

No other candidates received the support needed to win induction, in part because several of those players were also viewed as possible drug cheaters by some of the voters. The vote results underscored the lingering damage done by baseball’s steroid era, an extended period in which both baseball and its union stand accused of looking the other way while players used performance-enhancing drugs to gain an edge on the field.

But that era ended nearly a decade ago, and since then, Commissioner Bud Selig has gradually, and then more insistently, pushed for tougher testing. And the union has increasingly cooperated. After the Hall of Fame vote was disclosed Wednesday, Selig tried to make the best of the moment, saying he respected the writers’ decisions. Michael Weiner, the union’s executive director, disagreed with the writers’ verdict, calling it “simply unfair” that players like Bonds and Clemens had been denied access to Cooperstown.

Those differing reactions demonstrated that the two men have different constituencies to answer to. Still, the two have worked together to strengthen the existing testing program for several years now and the results are becoming increasingly apparent.

For instance, Major League Baseball was the first major sport in the United States to sign on to H.G.H. testing, reaching an agreement with its union in November 2011 to begin testing for the substance. It was a significant hurdle to clear because accepted H.G.H. testing requires blood samples, and baseball players, like any number of other athletes, had been reluctant to take that step after years in which drug testing was restricted to urine samples.

However, the 2011 agreement called for testing only in spring training and the off-season, reflecting concerns by the players about how their blood would be collected before or after regular-season games and whether the process would impact their performance on the field.

When the original agreement was announced, both sides said they would look into expanding the testing program for 2013, which they have now done. As a result, there will now be in-season testing for H.G.H., a substance that can help players build muscle mass and recover quickly from extended physical activity but which cannot be legally used without a prescription.

The agreement also establishes a testing regimen to detect abnormal levels of testosterone in a player’s body. In the last year, positive drug tests have linked a number of notable players — Ryan Braun of Milwaukee; Melky Cabrera, who was then with the San Francisco Giants; and Bartolo Colon of the Oakland A’s — to a high level of testosterone. Braun’s test result was overturned on appeal.

The new test will establish baseline levels of testosterone in players, creating a so-called longitudinal profile of each player’s testosterone ratio. Additional tests will be conducted on player samples that are considered outside their baselines to determine whether the player has used a performance-enhancing drug to increase the testosterone level.

In announcing the testing regimen, Selig stated that baseball would “continue to do everything we can to maintain a leadership stature in antidoping efforts in the years ahead.” Weiner, in his statement, said that “players want a program that is tough, scientifically accurate, backed by the latest proven scientific methods, and fair.”

So a day after the Hall of Fame vote, Selig and Weiner changed the subject and looked ahead, rather than keep gazing back at the rejected candidacies of some famous players.
 
Is it true that the 4:1 testosterone:epitestosterone ratio test is a bit of a joke amongst athletes and very easy to beat?
 
Is it true that the 4:1 testosterone:epitestosterone ratio test is a bit of a joke amongst athletes and very easy to beat?
Yes, athletes know how to "micro-dose" to avoid detection.

http://www.thehindu.com/sport/other-sports/article3710793.ece

Also, some athletes have genetic advantage when it comes to beating T:E ratio:

Testosterone:Epitestosterone Ratio Test - False Negatives and False Positives

Common Doping Test for Athletes is Unfair (and Racist)

Then, there's green tea:

Green Tea Helps Steroid-Using Athletes Beat Anti-Doping Test
 
Llouquet JL, Crepin N, Lasne F. The problem of anti-doping control of luteinizing hormone in boxing. Drug Test Anal. The problem of anti-doping control of luteinizing hormone in boxing - Llouquet - 2013 - Drug Testing and Analysis - Wiley Online Library

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. Anti-doping laboratories perform the assay of this hormone in urine and report abnormally elevated results. We observed a highly significant prevalence of abnormal results in samples taken after a boxing match. Comparison of the descriptive statistics for 426 LH values observed in boxing and other sports showed significant differences. An experimental study comparing urinary LH levels in 17 boxers before and after a match demonstrated a clear increase after the match. The same observation was made for urinary follicle stimulating hormone (FSH) in all of the eight boxers tested for this other pituitary gonadotropin. These observations have consequences for anti-doping controls, as the reference range for urinary LH levels must take into account the specificities of boxers. They also suggest consequences for the health of boxers. Although to our knowledge such observations have never been described, other pituitary disorders have been reported. Our results deserve further investigation from a medical point of view.
 
van der Gronde T, de Hon O, Haisma HJ, Pieters T. Gene doping: an overview and current implications for athletes. Br J Sports Med. Gene doping: an overview and current implications for athletes -- van der Gronde et al. -- British Journal of Sports Medicine

The possibility of gene doping, defined as the transfer of nucleic acid sequences and/or the use of normal or genetically modified cells to enhance sport performance, is a real concern in sports medicine. The abuse of knowledge and techniques gained in the area of gene therapy is a form of doping, and is prohibited for competitive athletes. As yet there is no conclusive evidence that that gene doping has been practiced in sport. However, given that gene therapy techniques improve continuously, the likelihood of abuse will increase.A literature search was conducted to identify the most relevant proteins based on their current gene doping potential using articles from Pubmed, Scopus and Embase published between 2006 and 2011. The final list of selected proteins were erythropoietin, insulin-like growth factor, growth hormone, myostatin, vascular endothelial growth factor, fibroblast growth factor, endorphin and enkephalin, alpha actinin 3, peroxisome proliferator-activated receptor-delta (PPARdelta) and cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C). We discuss these proteins with respect to their potential benefits, existing gene therapy experience in humans, potential risks, and chances of detection in current and future anti-doping controls.We have identified PPARdelta and PEPCK-C as having high potential for abuse. But we expect that for efficiency reasons, there will be a preference for inserting gene target combinations rather than single gene doping products. This will also further complicate detection.
 
This Table Summarises The Likelihood Of Each Protein Being Used For Gene Doping

For each protein, the average of the possible methods is considered (ie, the multiple options for inhibiting myostatin taken on average, just like the various FGF’s). If it is useful for athletes to abuse a protein, then it is marked in the first column as potential benefits. The experience in gene therapy-column has marks concerning the experience of gene therapy in humans.

When fully developed, some proteins remain more dangerous than others and this is noted in the third column, risk control. The chances of getting away with the illegitimate use of a gene-doping product with a specific protein are given in the column chance of undetected use. The last column indicates the likelihood of present abuse.

ACTN3, a-actinin 3; EPO, erythropoietin; GH, growth hormone; IGF, insulin-like growth factor; FGF, fibroblast growth factor; PEPCK-C, cytosolic phosphoenolpyruvate carboxykinase; PPARd, proliferator-activated receptor-d; VEGF, vascular endothelial growth factor.

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Djordjevic V, Stankovic I, Stipac AV, Putnikovic B, Neskovic AN. Short QT interval is unreliable marker of anabolic androgenic steroid abuse in competitive athletes. Srp Arh Celok Lek ;140(11-12):711-6. http://www.doiserbia.nb.rs/img/doi/0370-8179/2012/0370-81791212711D.pdf

INTRODUCTION: Previous animal and human studies provided the evidence that testosterone may affect ventricular repolarization by shortening of the QT interval. Synthetic derivatives of testosterone, modified to enhance its anabolic properties, are occasionally abused by some competitive athletes.

OBJECTIVE: We assessed whether the QT interval duration could discriminate androgenic anabolic steroids (AAS)-using strength athletes (SA) from drug-free endurance athletes (EA), by comparing 25 formulas for QT interval correction.

METHODS: We recruited 22 elite male athletes involved in long-term strength or endurance training and 20 sedentary controls. All elite SA reported AAS abuse, whereas EA and controls were AAS-free.

RESULTS: AAS-using SA had markedly shorter QT-interval than AAS-free EA (348 +/- 42.3 vs. 400 +/- 34.2 ms; p < 0.001). Also, drug-free EA had a significantly longer QT-interval than sedentary persons (400 +/- 34.2 vs. 358 +/- 18.9 ms; p < 0.01). In contrast, no significant difference in the QT-interval duration was observed between AAS users and control group (348 +/- 42.3 vs. 358 +/- 18.9 ms; p = 0.394). After the QT interval was adjusted for heart rate (HR) according to 25 different formulas, only the Ashman equation yielded considerable differences among the groups that were in line with those observed before correction.

CONCLUSION: Inconsistent results obtained by different correction formulas along with inability to discriminate QT (QTc) interval duration between AAS-misusing athletes and control group do not support the use of QT (QTc) interval for anti-doping purposes.
 
Gilbey A, Perezgonzalez JD. Health benefits of deer and elk velvet antler supplements: a systematic review of randomised controlled studies. N Z Med J 2012;125(1367):80-6. http://journal.nzma.org.nz/journal/abstract.php?id=5476 (NZMA: NZ Medical Journal)

AIMS: The aim of this systematic review was to evaluate the evidence from RCTs of velvet antler supplements for any condition, using the QUOROM statement as a guiding framework.

METHODS: Four electronic databases (PubMed, Medline, Web of Science and Academic search premier, via the bibliographical platform, Endnote) and two review articles were searched for all randomised clinical trials of velvet antler supplements. Retrieved trials were evaluated according to standardised criteria.

RESULTS: Seven RCTs were identified as satisfying all inclusion criteria and examined the effectiveness of velvet antler for rheumatoid arthritis (2), osteoarthritis (1), sexual function (1), and sporting performance enhancement (3). Their methodological quality ranged from 3-5, as measured on the Jadad scale. Two RCTs reported some positive effects of velvet antler supplements, but neither were convincing while the remaining five RCTs found no effect of velvet antler supplements.

CONCLUSIONS: Claims made for velvet antler supplements do not appear to be based upon rigorous research from human trials, although for osteoarthritis the findings may have some promise.
 
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