Michael Ashenden - Voice of Reason

[Galic Ho]

did you actually read my post?

I was defending the guy. Hence my first post in this thread .....





I have actually read the velo stuff - and yes I am quite well aware of who he is and how well respected he is thanks.

I have only ever posted very positive stuff about him, as I actually agree with you on pretty much all points. Not sure why you think I am not - but perhaps you are looking at another posters remarks???????

My apologies. My moment of madness. I misread your tone 100%. Next time I drop into the Clinic, I'll make sure I am fully awake and not in Zombie Land.

 

[Oldbiker]

Why is spiking the sample so difficult?

Several posters have stated that it is extremely difficult to spike the samples. I do not understand why it would be that difficult to make an EPO solution of a specific percentage and then taking a small amount( different for each vial) add it to a number of vials. Since LA had 17 samples. You would almost surely spike some of his during the process. If he had doped on one day wouldn't that days sample and the next also show EPO presence. So you might expect more than 6 of his samples to show positive.

 

[Special_oz_ed]

rhEPO vs concentration, and vs natural EPO

G'day Merckx,

I think we may have got our wires crossed.

Keep in mind that every urine sample is different. It doesn't contain a fixed amount of natural EPO, it might not contain a detectable amount. So even if you added a fixed amount of rhEPO to different urine samples, you would not get the same ratio of synthetic/natural in each.

Absolutely agree. Sorry but I think I wasn't very clear. I was talking about %rhEPO isoforms vs Concentration of rhEPO; that is, the ratio of rhEPO to natural EPO and also the concentration of rhEPO in the sample.

Here's a quote from Ashenden as to what it would take to spike the sample:

For the Prologue sample with 100% isoforms, the easy explanation would be that you would 'drown' the sample in EPO to ensure that it was 100%. Note - this is not failsafe, because compare sample 16 July (906 UI/L and 95.2%) against Prologue (600UI/L and 100%) and you see there is no correlation between EPO concentration and %isoforms!!!

http://velocitynation.com/content/features/2009/spiking-armstrongs-99-samples

It seems to me, from this example, to try to get a "random" looking mix of concentrations of rhEPO vs %isoforms it would take the most extraordinary of efforts, wouldn't it?

 

[Special_oz_ed]

Oldbiker,

Several posters have stated that it is extremely difficult to spike the samples. I do not understand why it would be that difficult to make an EPO solution of a specific percentage and then taking a small amount( different for each vial) add it to a number of vials. Since LA had 17 samples. You would almost surely spike some of his during the process. If he had doped on one day wouldn't that days sample and the next also show EPO presence. So you might expect more than 6 of his samples to show positive.

Not having investigated the lengths a lab would have to go to in order to spike a sample, and not being familiar with EPO analysis in urine, I at least considered the possibility that spiking could happen - I was still holding out hope LA hadn't doped - that is, until I found Ashenden's explanation.

BUT, if you check the Ashenden link I posted in reply to Merckx Index above (and in the quote below), then you'll see why I think spiking is next to impossible. It is very technical at points. A digest of it: to try to manipulate the samples to give you the kinds of % of EPO in the urine sample vs natural EPO and then to get the concentrations of EPO right would be next to impossible. The doses of EPO to spike the urine samples would be incredibly small - too small to be done by hand.

Here's Ashenden again,

Let's now try and manipulate that same sample to arrive at 95% basic isoforms. You would need .95 x 0.0042 UI = 0.00399 UI. At a concentration of 20,000UI per 0.6ml you would need 0.000000126 ml. This is 0.126 microlitres.

So 0.000000113 ml gives you 89.7% and 0.000000126 ml gives you 95%. Good luck finding a pipette able to get down to that level of accuracy so you could inject the corresponding amount.... Remember 1 microlitre is one millionth of a litre, so you want to get your accuracy down to less than 1 ten millionth of a litre.

And perhaps most importantly - remember that the smallest overdose in this context and the synthetic EPO will drown out the endogenous and you will get 100% basic isoforms (because literally their intensity will drown out any of the endogenous EPO appearing). http://velocitynation.com/content/features/2009/spiking-armstrongs-99-samples

An accuracy of less than 1 ten millionth of a litre is the telling part. That's simply not going to happen.

 

[Digger]

Several posters have stated that it is extremely difficult to spike the samples. I do not understand why it would be that difficult to make an EPO solution of a specific percentage and then taking a small amount( different for each vial) add it to a number of vials. Since LA had 17 samples. You would almost surely spike some of his during the process. If he had doped on one day wouldn't that days sample and the next also show EPO presence. So you might expect more than 6 of his samples to show positive.

Testers didn't know who the samples belonged to - they were anonymous. In order to make a guess correctly, it was a 480/1 chance they spiked the correct samples on each day.

 

[AussieGoddess]

and more they would have needed to know a lot about each sample they were spiking in order to get the balance right for a positive test ..... its not a case of simply adding a bit extra to a random few samples.

 

[Merckx index]

Absolutely agree. Sorry but I think I wasn't very clear. I was talking about %rhEPO isoforms vs Concentration of rhEPO; that is, the ratio of rhEPO to natural EPO and also the concentration of rhEPO in the sample.

When EPO is measured in the urine sample, it is total EPO, natural + synthetic. A distinction is not made; if it could be made, the whole gel process would be unnecessary. So the whole point is that in fact you cannot determine the ratio of bands to rhEPO amounts, or the ratio of amounts of rhEPO to natural EPO.

I think, though, there is an easier way to make this argument. Spiking a sample is not doing anything fundamentally different from actually doping, except the EPO is added to the urine after it is passed by the athlete, rather than before. Any pattern of bands and amount of EPO you could get from injecting EPO, and waiting till some of it got into the urine, you could also get from adding it directly to the urine. In both cases you are using a single preparation of synthetic EPO, so the relationship of the proportion of certain bands to total amount of rhEPO will be the same always. So even if you could measure these amounts—and again, you can’t—it would not preclude the possibility of spiking.

Let's now try and manipulate that same sample to arrive at 95% basic isoforms. You would need .95 x 0.0042 UI = 0.00399 UI. At a concentration of 20,000UI per 0.6ml you would need 0.000000126 ml. This is 0.126 microlitres.

So 0.000000113 ml gives you 89.7% and 0.000000126 ml gives you 95%. Good luck finding a pipette able to get down to that level of accuracy so you could inject the corresponding amount.... Remember 1 microlitre is one millionth of a litre, so you want to get your accuracy down to less than 1 ten millionth of a litre.

Again, you don’t need to add EPO from straight out of the bottle. You can dilute it first. So, in the example he gives, you could add 113 ul and 126 ul, respectively, of an appropriately diluted sample. The urine sample that is prepared for analysis is I believe around 20 ml, so adding that extra volume will not significantly affect the total sample volume.

Ashenden surely knows this. I think what he means is that the spiker would not bother to be that careful. He would just add a large amount of EPO to each sample, because after all, if 100% of the critical bands were observed, it would still be a positive. Why would he bother to add smaller amounts when it’s easier to drown the sample, as Ashenden puts it, and be certain of getting positives?

This is a reasonable point. But anyone familiar with the EPO test, as the spiker almost certainly would be, would know that there are maximum amounts of EPO possible in urine, even in dopers. He would not want to exceed that, because that would certainly suggest spiking. So he would add an amount of rhEPO that would stay well within those limits. And this amount might well result in band proportions of less than 100%, depending on how much natural EPO was present.

I think that at a certain point, Ashenden’s argument is like someone who sees some broken glass on his living room floor and says, that couldn’t have resulted from someone throwing a rock through the window, because if I did that, I could never replicate that exact pattern of broken glass. Of course one can’t, but the point is if one throws a rock through the window, one will get some highly improbable pattern of broken glass. In fact, the difference between 113 and 126 is about 10%, which could easily be pipetting error for someone who was not overly concerned with adding exact amounts. IOW, a spiker could have attempted to add about the same amount of rhEPO to two samples, and in fact added about 10% more to one than to the other. You couldn't "try" to add exactly the greater amount that was actually found, but it could certainly happen.

But as I said before, there are other arguments against spiking. I'm more concerned now with the statement Saugy made the other day, that their lab had evidence that upon long-term storage, EPO in urine was degraded in a way that resulted in more bands of the kind found in rhEPO. I'm a little astonished no one else has commented on this. When L'Equipe announced the positives in 05, several prominent anti-doping scientists, e.g., Ayotte, pointed out that it was extremely unlikely this could have resulted from degradation over time. If Saugy had evidence that this could happen, dating back to 02 and 03, why didn't he provide it?

If it were true, this would be a much better argument for LA's team. Spiking has a lot of problems, not the least of which is that someone who wanted to destroy LA's reputation had to know, at a minimum, that at least some of the samples used for research were LA's. And all the problems of adding the right amount of rhEPO, as discussed before. The degradation argument avoids all of that, and simply says as the samples sat frozen over the years, changes occurred producing bands that looked like those characteristic of rhEPO. It would also account for the positives in the other samples. The fact that a higher proportion of LA's samples were positive wouldn't matter much, because one could argue his urine might have more of the proteases which would catalyze this degradation.

I still think is very unlikely, though. Degradation of a protein will result in the protein becoming smaller. On a gel that determines size or molecular weight (SDS gel), this shift in size is very apparent. But the EPO test is based on differences in charge, not size (more recently, size separation has also been incorporated, but it was not used in 02-03, and even if it was, a change in charge would still have to occur for the patterns to look like rhEPO). While degradation could quite plausibly affect the charge of the bands, it is very unlikely that it would it produce bands looking just like the three major ones that are the key in the EPO test.

I think what Saugy is saying is that degradation results in a tendency for one or more of the bands from natural EPO to be converted into a more basic band, which could result in a significant bias in the test. IOW, if the existing pattern (before storage) was already fairly close to the 80% criterion, or whatever criterion happened to be in effect, the change might push it over the line. But even if this is the case, it is unlikely that a typical pattern from an undoped individual would be converted into a pattern characteristic of rhEPO.

 

[sashimono]

Your ire should be directed at sashimono who was actually attacking Ashenden...

It was just an opinion, not an attack.

If I was going to attack Ashenden I might have pointed out how in sports medicine "independent researcher" and "unemployed hack" are the same thing. Or I might have questioned why a "scientist" would spend so much time on public Lance-bashing, including attacks on Coyle et cetera before the 1999 EPO thing got his attention -- with no relevant data of his own to add to the discussion. But I didn't.

 

[ChrisE]

So let me get this straight....isn't this the same guy who was surprised at FL's admission of microdosing EPO?

Regardless, what matters is what can be proven. I am a broken record, admittedly. LA pis out of sight for 5+ years turns up positive with no established protocols in terms of checks/balances for the accused. No B sample. No witnessing of testing. No confirmation of chain of command, etc. Even if the feds retest the 99 samples alot of these same arguments can be made by the defense.

LA gives his codes.....that could be spun by by team LA by saying "why would he give the codes if he was dirty?" More doubt is sowed.

I'm not saying that was not LA's pis and it did not contain EPO as a result of him using it. I'm not saying that at all. I am just realistic on how this will play out IMO if it is part of the prosecution strategy.

 

[Merckx index]

So let me get this straight....isn't this the same guy who was surprised at FL's admission of microdosing EPO?

Yes, and I imagine this information might have spurred the investigation. This study was published very recently, and might well have been begun just after Ashenden heard about what Floyd had to say.

LA gives his codes.....that could be spun by by team LA by saying "why would he give the codes if he was dirty?" More doubt is sowed.

If one wants to play the spinning game, how about, LA gave the codes because he knew they had tested negative in the past, and he wanted to show the media he had nothing to hide. IIRC, didn't Ressiot get the numbers by asking LA something about a TUE? And LA answered forthrightly because he had a TUE, and wanted to show the press he wasn't hiding it.

I'm not saying that was not LA's pis and it did not contain EPO as a result of him using it. I'm not saying that at all. I am just realistic on how this will play out IMO if it is part of the prosecution strategy.

I think this is a reasonable point. The defense will certainly hammer home the fact that there was no B test, so that the evidence couldn't have been sanctionable. The fact that there was no proof of no tampering will make many people feel that the samples are out of bounds, that regardless of the arguments against spiking, no conclusions should be drawn about them. Also that they sat around so long. The defense, e.g., might ask, if you gave a blood sample that was stored frozen for several years, with no clear cut chain of custody, and then years later it tested positive for some serious disease, would you believe it, or would you insist on giving a fresh sample and being retested (which of course couldn't be done in LA's sample)?

I agree, points like this can sway a jury, no matter how strong the scientific evidence is. In this respect, I think Bert's case is somewhat similar. No matter how strong the evidence against contaminated meat is, a lot of people have trouble understanding how such a small amount of CB, appearing a day after a negative test, could possibly have been the result of intentional doping. It doesn't seem right.

But I still think the testimony of former teammates is what will make the case. The 99 samples, regardless of how strongly they are regarded as evidence, will certainly supplement the testimony, but I think that the testimony is critical.

 

[ChrisE]

....

If one wants to play the spinning game, how about, LA gave the codes because he knew they had tested negative in the past, and he wanted to show the media he had nothing to hide. IIRC, didn't Ressiot get the numbers by asking LA something about a TUE? And LA answered forthrightly because he had a TUE, and wanted to show the press he wasn't hiding it.



I think this is a reasonable point. The defense will certainly hammer home the fact that there was no B test, so that the evidence couldn't have been sanctionable. The fact that there was no proof of no tampering will make many people feel that the samples are out of bounds, that regardless of the arguments against spiking, no conclusions should be drawn about them. Also that they sat around so long. The defense, e.g., might ask, if you gave a blood sample that was stored frozen for several years, with no clear cut chain of custody, and then years later it tested positive for some serious disease, would you believe it, or would you insist on giving a fresh sample and being retested (which of course couldn't be done in LA's sample)?

I agree, points like this can sway a jury, no matter how strong the scientific evidence is. In this respect, I think Bert's case is somewhat similar. No matter how strong the evidence against contaminated meat is, a lot of people have trouble understanding how such a small amount of CB, appearing a day after a negative test, could possibly have been the result of intentional doping. It doesn't seem right.

But I still think the testimony of former teammates is what will make the case. The 99 samples, regardless of how strongly they are regarded as evidence, will certainly supplement the testimony, but I think that the testimony is critical.

I can't remember the details of under what circumstances the codes were given, but even if it was under the ruse of the TUE still LA is giving the keys of his samples to a journalist. I still think this can be spun by his lawyers as LA being clean is why he did it...he had nothing to hide.

As for AC, the rule is clear ie zero threshold. For this reason I do not see how he can get off, but nothing surprises me. If he gets off by CAS then that will be very damaging because it could be a slippery slope IMO; everybody will claim contamination in instances like these. I do think they need to do away with the zero threshold to get rid of the contamination argument for good, but this is a subject for another thread.

Yes, the testimony of others will be valuable, but remember there will be alot of other "Sally Jenkins" type characters in the sport, former teamates etc., that will testify he is the greatest thing since sliced bread and that they never saw him dope. I think alot of this scientific stuff as you say can be negated by a smart lawyer.

I am still unclear on whether or not it has been established if LA was an owner in Tailwind. He said in SCA he was I think, but then Herman or somebody denied it. If he wasn't an owner then what is he liable for? If that was the case he was no different than FL or TH, or GH.....

I also think, and I have said this many times, the lack of a formal AAF is very powerful. Read some of the comment sections after articles on espn or any sports website. This is a common theme touted by the LA believers, and it holds alot of weight with Joe six-pack regardless of how much we ridicule it in here.

 

[Merckx index]

Yes, the testimony of others will be valuable, but remember there will be alot of other "Sally Jenkins" type characters in the sport, former teamates etc., that will testify he is the greatest thing since sliced bread and that they never saw him dope.

A lot of LA fans are pointing out he had hundreds of former teammates, most of whom haven't ratted him out. I don't see how that helps much. The fact that someone never saw something isn't very good evidence that that something never happened.

The picture I get from Tyler is that LA and his most important lieutenants doped, and probably freely shared both substances and methods. It's less clear that others on the team were in on this. The guys who were not in the TDF, who might have ridden in other GTs or stage races or one day races, particularly as domestiques. I'm not saying they were clean, but if they weren't of direct benefit to LA, they might have been left to dope on their own. They wouldn't necessarily know what LA and the big boys on the team were doing.

I think it would be significant if a major lieutenant, someone who we would expect would be privy to everything guys like Tyler, Floyd and George were, came forward and said LA never doped. Ekimov comes to mind, but he has an obvious credibility problem, given his employer, and i don't believe he testified before the GJ (if he did, I'm very surprised he's commenting in public on Tyler's testimony). Assuming he did dope, and knows very well that Tyler is telling the truth, I think it's pretty sleazy to come out and deny. Given his job situation, I can understand keeping his mouth shut, but not going on the attack. I think it would be delicious irony if Eki were awarded Tyler's medal, then later evidence came out of his own doping. Apparently nothing short of threat of jail will get these guys to talk about what seems to have been common knowledge among much of the peloton.

Btw, Chris, I really miss your old avatar. He was so much better looking than the guy who immediately replaced him. But if you're into female journalists now, how about Bonnie Ford?

 

[Neworld]

When EPO is measured in the urine sample, it is total EPO, natural + synthetic

snip

But even if this is the case, it is unlikely that a typical pattern from an undoped individual would be converted into a pattern characteristic of rhEPO.

Excellent post Merckx. Thank you.

I wonder if Cobbles, Polish or Flick can digest, and I hope respond to, your fascinating value-added post?

NW

 

[AussieGoddess]

So let me get this straight....isn't this the same guy who was surprised at FL's admission of microdosing EPO?

Regardless, what matters is what can be proven. I am a broken record, admittedly. LA pis out of sight for 5+ years turns up positive with no established protocols in terms of checks/balances for the accused. No B sample. No witnessing of testing. No confirmation of chain of command, etc. Even if the feds retest the 99 samples alot of these same arguments can be made by the defense.

LA gives his codes.....that could be spun by by team LA by saying "why would he give the codes if he was dirty?" More doubt is sowed.

I'm not saying that was not LA's pis and it did not contain EPO as a result of him using it. I'm not saying that at all. I am just realistic on how this will play out IMO if it is part of the prosecution strategy.

You do make very good points here ChrisE

The tests alone are not sanctionable. The issues with the tests only being done for research purposes (and therefore perhaps looser chain of command, witnessing etc) and no B sample means that the UCI (or anyone else) were never going to be able to sanction LA on these tests alone.

That doesnt mean though that they are not good evidence.

They are still evidence pointing towards the conclusion that the feds are trying to reach - that LA used PED's. This evidence, in addition to the eyewitness testimony, the medical waste stuff from the 09 Tour, the finacial transactions, connections with Ferrarri, the stuff on Popo's laptop .... who knows what else.

These tests are not good enough on their own. But they are nonetheless useful towards building a case.

 

[rata de sentina]

These tests are not good enough on their own. But they are nonetheless useful towards building a case.

Well really the tests are good enough on their own. From a scientific perspective a number of independent longdituidinal samples positive is vastly superior to two aliquots of the same sample. The WADA rules are rigid because they are designed to maintain the integrity of a system but that doesn't mean that they are any more scientifically valid depending on the circumstances. The spiking scenario is so outlandish as to fall into the realm of one of those 9/11 conspiracy theories. I'm surprised that Ashenden even bothered with that demented drool.

 

[sashimono]

...The spiking scenario is so outlandish as to fall into the realm of one of those 9/11 conspiracy theories. I'm surprised that Ashenden even bothered with that demented drool.

I didn't read the original Lasne De Ceaurriz Nature paper, but the Vrijman report indicates that spiking samples was very much a part of the way they were building their database.

So its relevant, but obviously absurd to suggest that guys with their experience would fail to have meticulous records of what got spiked.

 

[lean mean &green]

Well really the tests are good enough on their own. From a scientific perspective a number of independent longdituidinal samples positive is vastly superior to two aliquots of the same sample. The WADA rules are rigid because they are designed to maintain the integrity of a system but that doesn't mean that they are any more scientifically valid depending on the circumstances. The spiking scenario is so outlandish as to fall into the realm of one of those 9/11 conspiracy theories. I'm surprised that Ashenden even bothered with that demented drool.

for the statistically challenged yes, 6 positive B samples are MORE conclusive than a single matching A/B sample. using a strict interpretation of WADA code it wasn't sanctionable but does it make good sense? umm, no.

merckx index's criticisms of ashendon are valid but i'd categorize them as academic. maybe even nit-picking when you consider he's talking to a cycling website, not presenting findings to a professional conference - word choice changes a bit. the layperson understands absolutes, not levels of confidence

spiking as an explanation is silly. the fact that such an explanation gained traction shows how easy it is to sway a fanatic. all that's needed to shift PR back the way you want it is willful ignorance and a sliver of doubt.

 

[rata de sentina]

I didn't read the original Lasne De Ceaurriz Nature paper, but the Vrijman report indicates that spiking samples was very much a part of the way they were building their database.

So its relevant, but obviously absurd to suggest that guys with their experience would fail to have meticulous records of what got spiked.

Spiking is useful in creating positive controls with known concentrations of the analyte especially in the initial stages of assay development when you might be uncertain of sensitivity. Also you might want to know what effect components of the urine would have on the analyte maybe during storage. You wouldn't spike unknowns but known negatives. The Vrijman report isn't a reliable source of information. I don't dispute the practicalities of spiking since it is a perfectly legitimate thing that you do in the lab when you are trying to get an assay working. In the context of the study of the 1999 samples it has no credibility.

 

[Merckx index]

The spiking scenario is so outlandish as to fall into the realm of one of those 9/11 conspiracy theories. I'm surprised that Ashenden even bothered with that demented drool.

merckx index's criticisms of ashendon are valid but i'd categorize them as academic… spiking as an explanation is silly

The problem is that a lot of what transpires in court rooms is demented drool. If these samples are actually entered as evidence, I would expect Fabiani to raise the spiking scenario. Really, it wouldn’t be any more of a fantasy than his other pronouncements. You can be sure LA is not just going to roll his eyes and say, I have no idea, but I never took EPO. He does have to give a reason why these positives could not be correct, and spiking and degradation are the only two alternatives that I see.

And spiking does not have to be intentional. How do we know that they were not accidentally contaminated? To someone not familiar with scientific labs, this might seem to be a possibility. Ashenden’s analysis is important in that it shows just why values like these could not result from some kind of accident. I would be very surprised if someone on LA’s team is not onto this already, trying to come up with some kind of explanation for these positives.

It’s easy to dismiss these possibilities now, but when the L’Equipe story first came out, a lot of people, including medical professionals, argued that lacking a clear chain of custody, these positives were meaningless. Someone will have to put this story to rest. There is too much hanging on these samples for there to be any doubts whatsoever.

 

[RTMcFadden]

Spiking is useful in creating positive controls with known concentrations of the analyte especially in the initial stages of assay development when you might be uncertain of sensitivity. Also you might want to know what effect components of the urine would have on the analyte maybe during storage.

As in, say, reasearch. Actually, this methodology can be used at three different points in the method life cycle: development, improvement, and validation.

You wouldn't spike unknowns but known negatives.

Like samples that have already been tested and deemed to be negative.

It's nice to see that after all of these years people have actually come to accept this reality.