Innovative Anti Doping Tests

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Haven't seen this program mentioned yet at the Clinic, but "Partnership for Clean Competition" has its podcast which features (among other topics) interviews now-and-then researchers who develop tests for anti-doping purposes such as a dried blood spots, OMICS approach to detect blood doping etc:


Very accessible and has some interesting topics, but I can't say I learned that much from the handful I've listened thus far, e.g. the AMGEN scientist who developed rHuEPO even mentioned something about having heard rumours of cyclists using the hormone in the late-1980s and it would've been interesting to know where the rumours originated from etc. instead of him spending minutes to tell the details of the ABP program.
There was evidence of rHuEPO use at both the Calgary & Seoul Olympics. Perhaps experimental use starting in the 1988 cycling season?

 
If we are to believe that what is written is true without taking a look into the sources quoted, then Francesco Moser was perhaps the first rHuEPO-user according to the CIRC-report (pp. 30-31):
Francesco Moser twice broke the hour record in 1984, reportedly helped by Dr Francesco Conconi and Dr Michele Ferrari. Francesco Moser is reported to have admitted using – at the time not illegal – blood transfusions in 1999, however in 2014 he apparently stated that: “…There wasn’t any blood transfusions, before the first or second attempt. The benefits of altitude were enough”. Seized documents from Dr. Francesco Conconi in proceedings against him apparently however showed that Francesco Moser had been treated with EPO around that time.
 
If we are to believe that what is written is true without taking a look into the sources quoted, then Francesco Moser was perhaps the first rHuEPO-user according to the CIRC-report (pp. 30-31):
Really is the wrong thread to be having this discussion again but: obviously Moser couldn't have used EPO in 1984. I think it's well known - as "known" as this can be in the face of omertà - that Moser's tenth-anniversary 1994 Hour rides were EPO-fuelled. That CIRC was inaccurate with its historical 'research' ... look, about the only good thing in that report is the way it highlighted how doping is defined by those talking about it, that for some it's puritanism for others it's simply not getting caught. For much of the rest, CIRC was badly written scuttlebutt.

WRT the EPO guy and the 'rumours' about its use in the 1980s. When did he say he heard these rumours? If in the 1980s, it's one thing, if later, well, there's lots of rumours about EPO use in the 1980s and this becomes like a self-referential Wiki claim eating itself. If it was contemporaneous rumours, well we know that EPO was being talked about in news reports from the Calgary Olympics. We know that Les Earnest was expressing an interest in it that same year. And we know that Søren Kragbak has said there was an attempt to recruit Dutch Danish cyclists in 1989. So, rumours of use in the 1980s: we know they exist.
 
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I have had only a precursory look at the article and haven't read it in too much detail yet, but here is a brand new article about a relatively new approach to detect autotransfusions:

https://cyclingtips.com/2018/08/game-changer-the-long-push-to-end-blood-transfusions-in-sport/

The research has been going on for a few years and while the approach has had some problems and there hasn't been (yet) a breakthrough, I know for sure that none-other-than Dr. Björn Ekblom (who "developed" blood doping in the late-1960's) is about 100 % certain that the method will eventually catch the blood dopers.
The breakthrough paper is going to be published relatively soon and a non-edited version is available here:


The authors claim that by focusing on differences between frozen vs. non-frozen RBCs, with their model, they can detect 100 % of autotransfusers (some for up to 5 weeks after the treatment) who use cryopreservation (-80 degrees C) with no false positives, when the control group has been sea level, high altitude and hypoxic chamber athletes.

According to the authors, this is an real breakthrough because they maintain that refrigerated blood is not a viable option for the following reasons:
Because cold-stored (+1º to +4ºC) blood is viable for a maximum of 42 days, during which time the athlete’s physical performance is compromised from blood donation, and erythropoietin is readily detected the blood doping method of choice for cheating athletes is transfusion of one´s own, long-term cryopreserved red blood cells.
Even when one should add ABP-fluctuations after blood donation roughly a month before reinfusion, I wouldn't be top convinced about this, because under the current ABP-system, even cryopreservation doesn't allow enormous transfusions without inducing fluctuations in Hb, Ret-% and OFF-Score.

In addition, an athlete can take certain precautions when donating blood the usual way not to affect the ABP-module or training capacity before the grand competition, such as:
  • elevating Hb mass via microdosing of cobalt/EPO before blood donation --> blood loss shouldn't affect ret-% count that much because the post-donation Hb-level is closer to normal than it would've otherwise have been.
  • Using the blood "refreshing" method (donate a bag, donate 2 bags a month later after which you infuse the old bag etc...) described by Hamilton, Landis et al, but only using something like ½ blood bags instead of a full bag --> Hb fluctuations affecting training status and ABP-module should be less than 5 %.
 
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The authors claim that by focusing on differences between frozen vs. non-frozen RBCs, with their model, they can detect 100 % of autotransfusers (some for up to 5 weeks after the treatment) who use cryopreservation (-80 degrees C) with no false positives, when the control group has been sea level, high altitude and hypoxic chamber athletes.
That's fantastic. Maybe, just maybe the days of blood doping are drawing to a close.
 
This is breakthrough research, but unfortunately freezing blood has also lost its relative advantage when one can't infuse 4-5 bags collected in October-February during a GT but instead must use a handful of 200-300 ml microinfusions that will likely survive in a non-frozen form.

Interestingly the arguments how Malm et al dismiss the viability of standard +4 C storage is very similar to that of Michael Ashenden in 2002 when -- despite his arguments -- blood dopers were many in number and used mainly their refrigerated (+4 C) blood (e.g. Freiburg+almost all clients of Fuentes+USPS) with a few exceptions.

Wrong in 2002, but hopefully not wrong today.

 
A little bit of wishful thinking at the moment but maybe a few years ago some of us would have said DBS testing was wishful thinking too:

View: https://twitter.com/RobinParisotto1/status/1288431094950293510


Parisotto's Tweet refers to an article titled 'Wearable Electronic Sweat Sensor Detects Health Biomarkers' the key takeaway from which is:
Engineers at Tufts University have shared their research in developing a flexible electronic sensing patch that analyzes sweat for a variety of biological markers, including sodium, ammonium, pH and lactate, directly from sweat and in a band format that can be sewn on to clothing.
The science bit:
The sensor platform is built as a patch where threads are placed on to the fabric gauze of a commercial bandage, the gauze enabling sweat transport from the sensing area on the patch to the reverse side of the patch for evaporation. However, the gauze edges are sealad to prevent sweat evaporating from the sensing area.

Sweat is wicked at the sensor surface for real-time measurement, and the threads connect to wireless readout electronics that permit real-time data acquisition and collection for detection and sensing of the biomarkers.
You can see what this looks like here. Basically, it's a Band Aid.



Change the markers being tested for and you can see where this could go. If it gets over the hurdles still ahead.
The study provides proof of feasibility, but lacks strict standardized measurements and statistically validated data at this point. The authors nod to this in their discussion write-up, and suggest that this will be the next step for development of the sensor: "The sensors can be statistically validated for their effectiveness in real-time continuous measurements of multiplexed biomarkers in the context of underlying clinical trials in lieu of sparse standardized measurements."
You can get your geek on with the full paper here.
 
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Perhaps you could summarise it for us...
  • a very sensitive method with no false positives.
  • WADA has the method under scrutiny with the reports available, but a Swedish anti-doping agency executice is optimistic.
I read like 3-4 articles about the method in the past 24 hours and not one mentioned that it only detects frozen blood, not the refrigerated one -- the latter one being cheaper, more accessible and fully viable for guys who don't need like 5-10 BBs during a season.
 
Thanks. So the usual hype without insight. Funny that WADA aren't on the hype train with it as well, though. I mean, they were promising us that our good friend Yannis Pitsiladis's mythical ground-breaking blood doping breakthrough was to have been ready for the Tokyo Games, which should have started a week ago.
 
So the usual hype without insight. Funny that WADA aren't on the hype train with it as well...
In defence of Malm et al, it at least looks a very good method in what it is doing, actually very good.

And it is "fair" in the sense that it'll make the llfe of former blood freezer-dopers (with a lot of money) - who donated blood during the OFFseason - more miserable when they have to use the same methods as their poor competitors.

Had it been the other way (detecting only refrigerated blood), a part of me would've found the method ethically dubious from frair-play perspective.

...If you are interested in coauthoring a short CN-article on the issue (I can do my part for free), I can write and DM you a memo on the history/application of the both of methods.
 
A little bit of wishful thinking at the moment but maybe a few years ago some of us would have said DBS testing was wishful thinking too:

View: https://twitter.com/RobinParisotto1/status/1288431094950293510


Parisotto's Tweet refers to an article titled 'Wearable Electronic Sweat Sensor Detects Health Biomarkers' the key takeaway from which is:The science bit:You can see what this looks like here. Basically, it's a Band Aid.



Change the markers being tested for and you can see where this could go. If it gets over the hurdles still ahead.You can get your geek on with the full paper here.
I'm really struggling to see how this could have anti-doping applications. Measuring ammonium, sodium etc. concs. in sweat compared to measuring metabolites of PEDs is very, very different. Developing a real time sensor to do this via skin sampling is even more different. There has been methods developed in the past that may be more applicable, this for example:


But even this would need to be tested to see if the metabolites of interest were actually seen, which is a very big if.
 
But even this would need to be tested to see if the metabolites of interest were actually seen, which is a very big if.
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:
Some biological markers have been detected in sweat for years so presumably many drug metabolites which 'do their work' in muscle tissue can in theory be transported (and detected) in the sweat that ensues from the tissue. Of course the research has to be done.
WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:
Dr. Rabin introduced the subject on the advantages and disadvantages of using doping control matrices different from urine and blood. He reminded that at present, only urine and blood were accepted matrices and that any other matrices could be used to support the results of an Adverse Analytical Finding but not to override it. WADA frequently received research proposals to use other matrices like saliva, hair, breath, sweat, and these were almost systematically not considered to be a priority. Examples included detection of clenbuterol in hair, perfluorocarbons in breath. Saliva was discussed by the HMRC in the past but did not have support due to the abundance of confounding factors. Sweat may be problematic due to contamination. Dried blood or urine spots were not considered to be alternative matrices since the biological fluids were the ones approved for doping control analysis. The questions posed to the HMRC was to whether WADA should give more consideration to alternative matrices, at least at the research level.

Several of the HMRC members gave their opinion. Prof. Handelsman believed there could be a place for the use of hair analysis. Prof. Ayotte mentioned that hair would only be useful for chronic administrations and if it was a matter of sensitivity, none of the other matrices would compare to urine or blood. Dr. Vernec noted it could be useful to distinguish the cases where a unique exposure was claimed. Dr. Barroso raised the fact that drugs stuck to hair differently depending on the colour and thickness. Dr. Wan noted that hair analysis is used in horse racing doping control, but even the follicle could become contaminated and there was no consensus on how to clean the hair. Dr. Pascual added that hair was always accessible to tampering. Prof. Ayotte proposed that WADA should convene a working group that should look into the published literature and recommend whether the results would be defendable in court. There was not much support for saliva: it could only be used for certain hormones but it could be easily contaminated with blood, and the collection required that the person didn’t eat for a certain period of time. Breath would be limited to a few substances and could be contaminated with saliva, which in turn could be contaminated with blood.
There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
 
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
Both papers you link are reviews, which is a bit annoying as it means reading all the references. Taking the second paper, that one also references another review in the sweat analysis section and one paper that looked at detecting the drugs you list in both saliva and sweat. The results are very mixed and they conclude in the abstract that saliva would be useful. They can't detect most of the drugs in sweat it seems (it's late so I've not signed in to get the full paper). The Tufts device wouldn't work to detect these things. Sweat or saliva might be useful, but it's going to be through sampling and analysing with something like the patch I linked to.
 

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