Marchand's B Sample test postponed

She is using the same strategy Lagat used. And the same strategy any OTC boy or girl will use if and when they get caught.

spaniel wrote:

You are just proving my original post. The EPO test separates by CHARGE, not size. If you are going to critique someone who has actually witnessed the test, perhaps you should at least read enough to understand it.

Whatever the physical property (I was assuming an SDS-PAGE gel was used - which is probably the case and measures proteins based on size), the point stands. It's incredibly unlikely that degradation would cause the size OR charge of natural EPO to change precisely to that of recombinant EPO. Thus it's incredibly unlikely that degradation would result in a false positive. Beating your chest about your personal observations of the test doesn't change the facts, but does make you look like a blowhard (many court experts are, sadly).

PS The test uses isoelectric focusing. For those interested in seeing the difference between a positive and negative test (at least circa 2006), see here:

http://bloodjournal.hematologylibrary.org/cgi/content/full/108/5/1778/FIG1

spaniel wrote:

You people should really stick to talking about things you have some sort of knowledge of. Why does the internet make people think they know everything?

Look, I have served as an expert witness for an EPO doping test. It paid for part of my wedding. So read and learn:.

There basically is no such thing as an expert (that isn't on the dark side), unless you are the sketchy scientists out there making the newer, harder to detect versions. You don't have any idea what conditions will change a drug test, or length of time at all. So while you may know what will happen with a drug sample of known out of date drugs, you are most certainly not up to date with anything relevant. You only know what information you've been given. And you haven't been given any information that would allow you to make such statements.

It's funny if you leave a sample for long enough a PED will always appear

5th in the NCAA XC is very good (plus top finishes in track)

Now we have a discussion, rather than what I have often seen as just an assumption; this is good.In your comment, there are enough different things that might be referred to by pronouns etc. that I find it a little uncertain exactly what you are saying and I want to clearly understand your comments. Would you kindly re-write this addressing the issues of time, heat, differences between the A and B results, etc. Repeating what I had said is that it is my understanding (see a long thread either here or T&FNews several years back with someone with access to relevant papers/tests posting) that Lagat's "A" sample was mistreated (kept over a weekend in a warm/hot environment (car?!) instead of being refrigerated) while the "B" sample was appropriated handled. I do not know how long after the event the "A" was tested and how long after the that test the "B" test was done. If you know anything in this regard, please add to the body of information on this matter.Thanks 26mi235

herr.doctor wrote:

Is IS commonly believed to have assisted Lagat. Feel free to dispute that it is not. I said nothing about "time matters", I stated what is a common opinion in this part of the world.

Reading is fundamental.

Hope you are not representative of the quality of education in Wisconsin.

26mi235 wrote:

Bullshit, Lagat's "A" sample was left for a long time in very hot conditions and almost certainly was degraded and damaged. The "B" sample was properly cared for, refrigerated etc. If you think that the time matters much look at the supposed positive of Armstrong's 1999 sample in about 2005 -- that is a longer interval by two orders of magnitude (although I will say that I do not remember how long it was between the test and the testing of either sample).

skinnie minnie wrote:

why can't her name be rachel instead of "racheal?" how pompus!

It's probably hispanic.

Logic wrote:

PS The test uses isoelectric focusing. For those interested in seeing the difference between a positive and negative test (at least circa 2006), see here:

http://bloodjournal.hematologylibrary.org/cgi/content/full/108/5/1778/FIG1

Great. Now look up and post Lagat's, and you will have a clearer understanding of the issue of degradation. What you linked is a clean sample.

I see. So being educated in the science behind the test, having conducted similar ones myself, and being qualified enough for an ATHLETE to trust me to verify the procedure does not make my opinion meaningful, but the ability to post on the internet makes you able to comment on this intelligently? Interesting logic.

spaniel wrote:

Great. Now look up and post Lagat's, and you will have a clearer understanding of the issue of degradation. What you linked is a clean sample.

What would Lagat's sample tell me? It's not a controlled sample like the ones I linked to. How many posts have you made now without addressing the real point? The fact that you're so interested in appealing to your own authority (a logical fallacy) indicates to me that you just can't admit that you're wrong.

Here's the point again in case you ever decide to post something with substance:

It's incredibly unlikely that degradation would cause the size OR charge of natural EPO to change precisely to that of recombinant EPO. Thus it's incredibly unlikely that degradation would result in a false positive.

I won't pretend to be particularly knowledgeable but it looks like a positive is defined by the bands falling outside a defined "endogenous" range. So any alteration in net charge could potentially create a false positive.

Logic wrote:

What would Lagat's sample tell me? It's not a controlled sample like the ones I linked to. How many posts have you made now without addressing the realp oint? The fact that you're so interested in appealing to your own authority (a logical fallacy) indicates to me that you just can't admit that you're wrong.

Here's the point again in case you ever decide to post something with substance:

It's incredibly unlikely that degradation would cause the size OR charge of natural EPO to change precisely to that of recombinant EPO. Thus it's incredibly unlikely that degradation would result in a false positive.

Holy wow are you obtuse. You don't think Lagat's sample was run with a control lane on the gel??? Then what on earth did they compare it to? Of course it was a controlled sample! Once again, just like "assuming" it was SDS-PAGE when it was not and then blithering on about a test you did not even understand, you are speaking about things you know nothing about. If you had actually looked up Lagat's gel image, it would be very clear to you how ambiguous these tests can be, especially with a degraded sample.

To add to the complexity, there is plenty of evidence that the isotype distribution of endogenous EPO varies between individuals and is altered by training. So two people's EPO may be charged differently and appear differently on a gel (with some of it appearing in the "doped" area), and a person's endogenous EPO may appear differently depending on their training. These are well-known criticisms of the test, as well as the fact that they use a monoclonal antibody for detection with is NOT specific to exogenous EPO and they run a 1-D test on charge instead of a 2-D test on charge and molecular weight.

You say "it's incredibly unlikely" but this is simply your completely uninformed opinion and not based on anything. If you had read Heid's report on Lagat it would be obvious to you how it is not that simple. But I suppose it is easier to make a fool of yourself than educate yourself.

for what its worth wrote:

http://www.wada-ama.org/rtecontent/document/td2004epo_en.pdf

I won't pretend to be particularly knowledgeable but it looks like a positive is defined by the bands falling outside a defined "endogenous" range. So any alteration in net charge could potentially create a false positive.

Yes, they take steps to lessen the likelihood that a simple shift will be misread but as you imply it is the bands falling in an area, whether than the precise position, which defines a positive. And thank you for adding real information.

No Lagat's sample is not controlled like the ones I linked to. His sample was experimental - that's the whole point of the test. If his sample was a control then there wouldn't have been any point to running the sample. A control is sample that gives a known result. Talk about obtuse. You've spoken in a court of law as an expert? Seriously?

Talk about blathering! You sure can type a lot of words without saying anything. Until you show a controlled experiment that shows a sample that is known to have undergone degradation (not just speculation), you haven't addressed the point.

What does this have to do with the effect of sample degradation on the test?

I have educated myself. But no thanks to you. From your first post you've been completely arrogant, rude, and obnoxious and you're contributions to the thread have been minimal except to brag about how smart you are. You must be real fun at parties.

for what its worth wrote:

http://www.wada-ama.org/rtecontent/document/td2004epo_en.pdf

I won't pretend to be particularly knowledgeable but it looks like a positive is defined by the bands falling outside a defined "endogenous" range. So any alteration in net charge could potentially create a false positive.

My understanding is that it's not the presence of ANY band outside the endogenous range, which I agree would be more prone to false positives, but the presence of bands in a specific area that gives a positive. From the test literature, "in the basic area there must be at least 3 consecutive bands..."

To abstract from this document:"11. Summary A) All experts present agreed that the urine of Mr. Bernard Lagat did not contain any recombinant erythropoietin (rhEpo) and he therefore had not committed Epo-doping.B) While the test of Mr. Lagat´s B-sample did not reveal any reactive components in positions taken as indication of a positive test, the original A-sample showed bands in this region which, however, were not fully identical with those of the reference recombinant Epo and might well be due to glycoprotein modifications. C) The test used involves many steps with many pitfalls. Only very experienced and critical “bench biochemists”, as those working in the Cologne lab, can responsibly handle the complicated and error-prone procedure. There are several problems with background staining, reproducibility, technical artifacts, losses of proteins during the enrichment steps and losses and changes as result of glycoprotein modifications etc."more Letters, mainly about the issues of the test and its attributes/difficulties etc.

Bob Dobbs wrote:

http://www.google.com/url?sa=t&source=web&cd=1&ved=0CBgQFjAA&url=http%3A%2F%2Fwww.letsrun.com%2F2003%2Flagatfull.doc&rct=j&q=%22Report%20of%20B-sample%20testing%20in%20the%20laboratory%20of%20Prof.%20W.%20Sch%C3%A4nzer%2C%20Institute%20of%20Biochemistry%2C%20Germany%2C%20Sport%20University%2C%20Cologne%22&ei=-kuZTLaMNsb_lgfd1owO&usg=AFQjCNGZwOOXyDljBNwEfUfQ_hhj0McQIA&sig2=qYf79gJHMnspPwrmHTfx6g&cad=rja

Logic wrote:

"No Lagat's sample is not controlled like the ones I linked to. His sample was experimental - that's the whole point of the test. If his sample was a control then there wouldn't have been any point to running the sample. A control is sample that gives a known result. Talk about obtuse. You've spoken in a court of law as an expert? Seriously?"

Any EPO test is a controlled test. There are both experimental samples and control samples run. One of the fundamental problems with Lagat's test, and ALL EPO test run prior to that, was that they did not control for degradation. It was not until Heid's scathing report that a degradation control was added. That said, if you would simply look up Lagat's test you would easily see that Lagat's test lane, and a known positive, do not appear identical at all.

If a recombinate-specific monoclonal antibody were used, a false-positive would not result due to degradation, only a false-negative would be possible. If a 2-D process rather than a 1-D process were used, degradation resulting in a charge shift would not result in a false positive unless it also resulted in a matching weight shift, so the odds of a false-positive would be greatly reduced. Yet ten years after this test was developed neither is done.

While protein stored at -20C will be stable for several years and that stored at -80C for many more, instability may result upon thawing if the sample is not properly maintained during testing. While protease inhibitors are added to the sample, these chemicals do not necessarily inhibit glycosidases, which are capable of removing the sugar groups which, to a large extent, determine the charge differences between endogenous and exogenous EPO. A standard and rational step to take to reduce the likelihood of this would be to run the samples in refrigerated centrifuges and other equipment, which would inhibit all degrading enzymes to some extent. Yet despite taking over 48 hours to run this test through all of its many steps, it is run at room temperature despite the equipment being capable of refrigeration. This is yet another factor which may result in uncertainty in the results if degrading enzymes are present.

"Talk about blathering! You sure can type a lot of words without saying anything. Until you show a controlled experiment that shows a sample that is known to have undergone degradation (not just speculation), you haven't addressed the point."

Multiple experts agreed with me, hence Lagat was cleared. How can you, a random anonymous internet poster arguing about a test they don't even bother to understand is not SDS-PAGE before running their mouth, pretend to know better?

"What does this have to do with the effect of sample degradation on the test?"

If you were educated enough on protein biochemistry to understand what I had written, the connection would be obvious. I would explain further but you have made if very clear that you are interested in arguing, not learning reality.

"I have educated myself. But no thanks to you."

Unlike other posters here, who actually looked up the test and provided useful information you could learn from, you have not brought in any factual information.

" From your first post you've been completely arrogant, rude, and obnoxious and you're contributions to the thread have been minimal except to brag about how smart you are. You must be real fun at parties."

Stating actual experience with the test and a knowledge base to comment from is not arrogance. But I do not suffer fools well, you are right for once. While it has been entertaining in a sad way, I think I'm done here, this is the reason nobody reads these boards anymore.