floyd's tests, discuss the science, influences... Options

[Bill Hue]

Catchy Carbon; November 2006; Scientific American Magazine; by Sarah Simpson; 2 Page(s)

When marine chemist John Hayes pioneered a way to scrutinize the carbon atoms in seafloor mud 15 years ago, he was trying to unravel mysteries about how dead microbes once lived. Never did he guess that sporting officials would one day use his invention to catch drug cheats.

By modifying Hayes's method to examine the carbon atoms in athletes' urine, medical researchers at the U.C.L.A. Olympic Laboratory have developed the first definitive screen for synthetic testosterone, a popular anabolic steroid banned by most sports organizations since the 1970s. The new test--known as the carbon isotope ratio (CIR) test--figures prominently in several recent, high-profile doping cases, including the disqualification of sprinter Justin Gatlin's world record in the 100 meters. It may also strip cyclist Floyd Landis of his 2006 Tour de France title.
."...if the body were able to make testosterone from an artificial compund-such as the cortisone athletes sometimes inject to reduce muscle inflammation-might the natural hormone carry a synthetic-looking finger-print." Hayes notes.


So, can the body make testosterone with a sythetic looking fingerprint from cortisone? Is that then the an "internal" or natural explaination for the "external" CIR result?



The metabolites of cortisol are among those metabolites used by WADA as references to give a baseline of comparison with the testosterone metabolites. In other words, the cortisone he was taking didn't convert somehow to testosterone. His (possibly synthetic) cortisol metabolites would be compared directly to his testosterone metabolites.
Thomas A Fine


Does this explain the ratio differential?


This is hard stuff for me.
Bill

[floyd]

Catchy Carbon; November 2006; Scientific American Magazine; by Sarah Simpson; 2 Page(s)

When marine chemist John Hayes pioneered a way to scrutinize the carbon atoms in seafloor mud 15 years ago, he was trying to unravel mysteries about how dead microbes once lived. Never did he guess that sporting officials would one day use his invention to catch drug cheats.

By modifying Hayes's method to examine the carbon atoms in athletes' urine, medical researchers at the U.C.L.A. Olympic Laboratory have developed the first definitive screen for synthetic testosterone, a popular anabolic steroid banned by most sports organizations since the 1970s. The new test--known as the carbon isotope ratio (CIR) test--figures prominently in several recent, high-profile doping cases, including the disqualification of sprinter Justin Gatlin's world record in the 100 meters. It may also strip cyclist Floyd Landis of his 2006 Tour de France title.
."...if the body were able to make testosterone from an artificial compund-such as the cortisone athletes sometimes inject to reduce muscle inflammation-might the natural hormone carry a synthetic-looking finger-print." Hayes notes.
So, can the body make testosterone with a sythetic looking fingerprint from cortisone? Is that then the an "internal" or natural explaination for the "external" CIR result?
The metabolites of cortisol are among those metabolites used by WADA as references to give a baseline of comparison with the testosterone metabolites. In other words, the cortisone he was taking didn't convert somehow to testosterone. His (possibly synthetic) cortisol metabolites would be compared directly to his testosterone metabolites.
Thomas A Fine
Does this explain the ratio differential?
This is hard stuff for me.
Bill

Thanks Bill, I don't really understand it but it seems to me that they should never use metabolites of cortisone until they have ruled out the possibility that they may contain exogenous molecules.

[Bill Hue]

Thanks Bill, I don't really understand it but it seems to me that they should never use metabolites of cortisone until they have ruled out the possibility that they may contain exogenous molecules.

And they have NOT ruled that out. It will be interesting to see how the panel deals with that.
Do you happen to know their burden of proof or do you have the burden?

[floyd]

And they have NOT ruled that out. It will be interesting to see how the panel deals with that.
Do you happen to know their burden of proof or do you have the burden?

I'll research that, but it sure feels like I have it.

[Bill Hue]

I'll research that, but it sure feels like I have it.

If you do, then you have the obligation to forward the theory- or they will dismiss it like they always do. If the national federation has the proof, then forwarding the theory calls their contention into question.
The literature is not real clear that the cortisone metabolite can make a sythetic looking fingerprint.

Thomas A Fine's notion though, about the ratio, if that is what it is a little more scientifically developed, I think???

[floyd]

If you do, then you have the obligation to forward the theory- or they will dismiss it like they always do. If the national federation has the proof, then forwarding the theory calls their contention into question.
The literature is not real clear that the cortisone metabolite can make a sythetic looking fingerprint.

Thomas A Fine's notion though, about the ratio, if that is what it is a little more scientifically developed, I think???

Thanks, I'll tell the "experts" about that.

[Bill Hue]

Your welcome. I'm sure none of this is new to your team.
Bill

[MacRoadie]

Thanks, I'll tell the "experts" about that.

As an architect, I'd like to think I bring a reasonably balanced left/right brain approach to problem solving (a kind of holistic approach to rational analysis). As an expert witness in building forensics, I am not unfamiliar with the process of discovery and the rather "black and white" approach taken within the legal arena.

What I find both fascinating and disturbing about this whole matter is this apparent dichotomy between the empirical nature of the testing and what would appear to be a quite subjective process of interpreting the test results.

Many more knowledgeable posters have provided us with a rather thorough description of what to the lay person would appear to be a very exhaustive laboratory process. We're not talking about simply dipping some litmus paper in a cup or peeing on an early pregnancy test strip.

In stark contrast to the empirical process of lab work, we are then confronted by innumerous interpretations of the lab results (setting aside for a moment any question or theory of lab error). Many of these are based on solid science, some are possibly wild or speculative in nature.

The bottom line seems not to necessarily be what the test results are (although those are obviously subject to dissection), but how they were achieved (again, not suggestive of lab error) be it through natural metabolic processes, or some other phenomena.

In my arena, we as experts can agree to disagree with regard to interpretation of our investigative analysis. We can look at a condition and come to 4 or 5 logical and plausible explanations for it's existence, all relying on the same physics and mathematical formulae. We can agree that there can be multiple contributing factors to an assembly's failure, without agreeing on causation.

We have that "luxury". I don't think Floyd does and this is what concerns me most of all.

[adapa]

Catchy Carbon; November 2006; Scientific American Magazine; by Sarah Simpson; 2 Page(s)


."...if the body were able to make testosterone from an artificial compund-such as the cortisone athletes sometimes inject to reduce muscle inflammation-might the natural hormone carry a synthetic-looking finger-print." Hayes notes.
So, can the body make testosterone with a sythetic looking fingerprint from cortisone? Is that then the an "internal" or natural explaination for the "external" CIR result?

The metabolites of cortisol are among those metabolites used by WADA as references to give a baseline of comparison with the testosterone metabolites. In other words, the cortisone he was taking didn't convert somehow to testosterone. His (possibly synthetic) cortisol metabolites would be compared directly to his testosterone metabolites.
Thomas A Fine
Does this explain the ratio differential?
This is hard stuff for me.
Bill

This doesn't seem correct to me but I'm a DALP also....

I'm a bit leary of accepting the statement by Hayes that the 'if the body were abel to make T from Cortisone' as a possability. If I'm reading the quote correctly it's from 1990 so I would assume there should have been some evidence since then to back the statment up.

RH did make a reference to doing a lot of research on this subject, I can't find the postings at this point but quote his reference to it.
Also, I do *not* fully understand what he's stating so can not comment on it.

"In brief,

1. Cortisone shots do not result in metabolic intermediate compounds that after leaking into urine could be confused with testosterone metabolites as seen by the specific test - IRMS - isotope ratio mass spectrometry. I researched the subject for hours after I saw the statement in the media.

2. IRMS determined presence of exogenous testosterone, not exogenous cortisone. There can be no confusion of two products. The amount is irrelevant.

3. Cortisone shots do not typically change T/E ratio. Cortisone is a biochemical antipode of testosterone.
If anything it would tend to LOWER testosterone and T/E.

4. Test error is of cause possible but I doubt it. The test was adapted in 1999. That why Botero just barely got away. This is the exact same test that caught Gatlin despite what media reported.

5. Anabolic steroids and testosterone are NOT tested in the same way. They are only screened out by the same test. Once the flag is raised they undergo different tests."
http://www.dailypelotonforums.com/main/ind...topic=831&st=60 post #72

This post has been edited by adapa: Oct 11 2006, 03:09 AM

[Thomas A. Fine]

Floyd asks about cortison metabolites being used as references...

Floyd, It's a good question and I know that certain studies I saw used cortisone metabolites or a mixture of
those with some other ERCs. But at the end (as I just explained in the Floyd thread to Vaun's question) they'll mostly like make a conclusion on the basis of "likely ERC compared to "likely" metabolites. For example androstane compared to pregnane. Cortisol may be ignored in results because it would be used for cortisone like metabolites.

Rational, lets assume for a second that, since it is Floyd, this question wasn't just a stab in the dark. I mean, he may not know the science, but he knows what was tested. And metabolites of cortisone are valid ERCs. And additionally I'll put out that in the past people have said that the lab would be stupid to use a metabolite of a TUE for an ERC, but everybody including me failed to remember something - when the lab does the tests, they (in theory) don't know who they're testing for, so there's no way to eliminate a TUE
metabolite before the fact.

So let's just answer the question.

Unfortunately Floyd, it doesn't skew your way. If you still have synthetic cortisone in your system, and it's metabolites are used as a reference, then this will skew the test towards masking a positive, because the synthetic cortisone would have carbon-13 close to that of synthetic testosterone. Synthetic cortisone would actually be a protection against failing thist test.

Of course, if a cortisol metabolite was used as a reference, and you still had synthetic cortisone in your system, and you ended up testing positive, this would be what is technically referred to as "whacked". If the synthetic cortisone didn't mask out a (true or false) positive, then the measurements for the reference would be highly suspect. I guess in that sense it might skew your way.

This is all assuming that synthetic cortisone is made from soy or yams (the best info I've seen suggests that this has been the case in the past). If it turned out that synthetic cortisone is made from corn, then this would tend to create a false positive.

Did you have synthetic cortisone in your system? The half-life for the pure form is very short, but they probably inject you with something that metabolizes slowly, much as they do with weekly testosterone injections.

Speaking of which, what about longitudinal studies? The WADA protocol looks to me liike it says that longitudinal studies are required, if available. This should mean that you should have isotope test results from your piss two days earlier and two days later.

tom

[dbrower]

I'll research that, but it sure feels like I have it.

Man, I just spewed my drink on my screen with that one.

I can just imagine a Leslie Visser interview: "Tell me Floyd, how does it feel to have the burden of proof on your shoulders? Is it heavy? Does it hurt? Have you trained for it?"

-dB

[cyclenut]

Does synthetic thyroid play any part in any of this, by chance? Or do you take animal thyroid instead of synthetic? And if you take animal, are there any chances of products the animal was given (growth hormones) could show up in your test results?

And all that water you poured over your head in S17, are there any products that leach out of the plastic? Just a thought....

[dbrower]

If you do, then you have the obligation to forward the theory- or they will dismiss it like they always do. If the national federation has the proof, then forwarding the theory calls their contention into question.
The literature is not real clear that the cortisone metabolite can make a sythetic looking fingerprint.

It's way worse than that -- he can't just come with a theory, he needs to come in with some pretty unassailable test results that demonstrate the theory.

The only case I've found where there was clear proof was one I titled, "You can get off -- if you are a horse with cancer". In this one, the horse had a whopping TE, which the defense claimed was from an undiagnosed cancer; when removed, the levels returned to normal and CAS accepted it was endogenous.

see http://www.horsesport.org/PDFS/FEI/05_02/D...mine_DAuzay.pdf

-dB http://trustbut.blogspot.com for Landis news, research, and comment.

[MacRoadie]

Man, I just spewed my drink on my screen with that one.

I can just imagine a Leslie Visser interview: "Tell me Floyd, how does it feel to have the burden of proof on your shoulders? Is it heavy? Does it hurt? Have you trained for it?"

-dB

I'm imagining Kirsten Gum, apres "enhancement" of course...

[rational head]

Floyd asks about cortison metabolites being used as references...
Rational, lets assume for a second that, since it is Floyd, this question wasn't just a stab in the dark. I mean, he may not know the science, but he knows what was tested.

Tom, we discussed this issue extensively. One-mint and Hopar are quoted above. When I am asked a question I try to answer it as if I asked something I don't know much about. Bottom line, little probability of cortisone metabolites messing up the results..But I'm only judging from what I've seen in the studies.
---------
Below is a bunch of general and more specific interesting studies on Cortisone and more I checked out. May be Floyd or anyone else will find some help there.
----------------
Regarding T/E longitude study, check WADA papers - it's a fall back at best -they are described very well.
----------
Negative relationship btwn T and cortisol

http://www.jssm.org/vol4/n1/10/v4n1-10pdf.pdf

Same

http://www.ncbi.nlm.nih.gov/entrez/query.f...5&dopt=Abstract

Cortisol and T.

http://www.exrx.net/AnabolicSteroids/Testosterone.html

Cortizon/T influences in various excersize modes

http://www.the-aps.org/press/journal/28.htm

Cortisol byproducts

http://lib.bioinfo.pl/pmid:16183337


Cortisol and thyroid

25. Ueki M, Okano M. Analysis of exogenous dehydroepiandrosterone excretion in urine by gas chromatography/combustion/isotope ratio mass spectrometry. Rapid Commun Mass Spectrom. 1999;13(22):2237-43.

Cortisol in urine

http://www.endocrine-abstracts.org/ea/0002/ea0002P97.htm

Pfizer's Prednisone data (what it's made of)
http://www.pfizercentresource.com/product_...ocortisone.html

Vegetable Origin of Raw MaterialsPfizer produces steroid active pharmaceutical ingredients (APIs) by what is best described as a semi-synthetic process using a crude mixture of vegetable sterols that are isolated from various oilseeds as the starting material. These vegetable sterols, stigmasterol and sitosterol, are processed through several fermentation and chemical steps to yield Hydrocortisone Hemisuccinate.




109 studies of T and various metabolic factors

http://www.ncbi.nlm.nih.gov/entrez/query.f...Pager&DB=pubmed

Study of false positives

http://www.ncbi.nlm.nih.gov/entrez/query.f...2&dopt=Abstract

False positives

http://www.thesportjournal.org/2002Journal...o1/drug-use.htm

T metabolites

http://www.ncbi.nlm.nih.gov/entrez/query.f...1&dopt=Abstract

[dbrower]

"I can just imagine a Leslie Visser interview..."

I'm imagining Kirsten Gum, apres "enhancement" of course...

Cheeze, I'm showing my age. Whose this Kristin Gum, young feller? Wasn't she with Vance Armstrong before running off with a Wrigley heir?

-dB

[jr.]

Tom

I find the WADA Technical Document a little ambiguous in that Section 4, Reviewing and evaluating test results. contains bullet points without indicating conjunctive/disjunctive or other hints as to the intended construction. But when read together with the previous section that sets out the standards for a 'finding consistent with administration of a steroid' versus a finding which should be reported as "inconclusive" and looking at the flow chart in Guideline/Reporting and Managing of Elevated T/E ratios, it appears to me that no longitudinal study would be required of the lab or testing authority if the initial analysis is that the metabolites measured three delta units or more from the urinary reference steroid or if the c13/c12 value can't be measured but the ratio for the metabolites is below -28 0/00. If the tests don't fall into the conclusive category, that's when the longitudinal studies described in the technical document would be performed.

I don't work for WADA, so I won't guarantee my analysis, but I think that is how its applied. I suppose Thursday we will all see if there are IRMS on the other samples FL submitted at the Tour in the lab report. As a matter of curiosity I would like to see IRMS results on all of his samples. While almost anything can happen, no matter how stupid it appears to outsiders, ie, the no one would take testosterone for one day theory, it would seem unlikely that all his other samples would show up as inconclusive if FL were on a typical doping program.

[weezy]

here's a thot -- everyone is looking at FL's results on one day -- can the defense (FL) ask to see ALL tests of all riders for the entire tour -- or even the entire season -- to see where his "alleged" results fall with respect to all tests administered? and how those results were interpreted? and how they were handled?

if he got access to them, they would most likely have to be anonymous -- and the larger sample of multiple races would make breaking a code as to whose was whose less likely, which would reduce invasion of privacy concerns.

can/should FL request all of his raw results and interpretations from all tests for the year? from whatever lab did the testing?

(it just occurred to me that this might be THE all-time record holder for number of people thinking about, and providing input for, an arbitration.

[Thomas A. Fine]

A long time ago, in a posting that started this very thread...

2. IRMS test

By now everyone probably is clear on how it works - so I'll spare my time here.
I re-reviewed all my references. I think it's a very robust test. The instrumentation is difficult to calibrate but it's plenty accurate. By far, the best reference material is this
If you want to understand EVERYTHING about the test -sample preparation, metabolites, statistical analysis, qualification criteria, nutritional, gender and race influences - it's all there. It clearly shows why 3 SD are sufficient. It answers earlier questions about the actual as found SD and why they are low. It explains why SD in other studies were higher.

First, I think it's pretty clear that very few people understands how this test works, and that it can be repeated often for the sake of the newcomers, and those who keep glossing over the details.

At any rate, that reference is interesting. Look at Figure 2. 5betaP (which is a common reference compound) is on average about 2 per mil higher than either of the other two compounds tested (which are testosterone metabolites). Doesn't this mean that all of these 73 normal subjects in the control group have a two per mil head start towards a positive? In other words, one of their T metabolites would only have to vary by one per mil negative in order to result in a postive test? That's less than 1.5 SD
variation.

Look at Table 3. This has various stats about the control group. Look at the line marked "Maximum". This shows the maximum per mil values of the tested metabolites. It also shows the maximum differences. Look closely at this -- the differences are not the differences between the maximums listed, that doesn't add up. They must actually be the maximum differences found in the control group. These are postives for doping. Since it is a control group, presumably they're false positives.

Jeez, am I going to find false positives in every "supportive" study I look at?

tom

[Will]

Nice work, Tom.