Doping inspector backs Armstrong

[Anonymous]

And one liner put downs!!!!

Yea, I am not sure that would endear me to a jury, but I might get a laugh or two...and contempt of court jail time like Hugh said. I can picture the "How funny do you feel now Mr Smartass" looks from the judge when the bailiff leads me out...

 

[CentralCaliBike]

Yea, I am not sure that would endear me to a jury, but I might get a laugh or two...and contempt of court jail time like Hugh said. I can picture the "How funny do you feel now Mr Smartass" looks from the judge when the bailiff leads me out...

State judges generally will give you a little latitude (especially if you go to the dark side) but the Feds have no since of humor at all.

If you are looking to get in the courtroom, I think you will find that the local county prosecutor's office will give you a lot more experience. The larger offices have civil consumer protection units. If you are more interested in the prosecution side most also have welfare fraud, white collar (usually embezzlement type cases, but also includes large scale check forgery operations), worker's compensation fraud, and real estate fraud.

Of course if you want to stay with your personality and get in the one liner or two you might consider prosecuting violent crime - juries are perfectly ok with you verbally beating up on a murderer or rapist where they might not be interested in hearing that great slam for the woman who misstated the number of days the ex lived in the house causing a $1000.00 overpayment of welfare.

 

[Mellow Velo]

State judges generally will give you a little latitude (especially if you go to the dark side) but the Feds have no since of humor at all.

Must mean they are all Armstrong fanboys.

 

[Anonymous]

State judges generally will give you a little latitude (especially if you go to the dark side) but the Feds have no since of humor at all.

If you are looking to get in the courtroom, I think you will find that the local county prosecutor's office will give you a lot more experience. The larger offices have civil consumer protection units. If you are more interested in the prosecution side most also have welfare fraud, white collar (usually embezzlement type cases, but also includes large scale check forgery operations), worker's compensation fraud, and real estate fraud.

Of course if you want to stay with your personality and get in the one liner or two you might consider prosecuting violent crime - juries are perfectly ok with you verbally beating up on a murderer or rapist where they might not be interested in hearing that great slam for the woman who misstated the number of days the ex lived in the house causing a $1000.00 overpayment of welfare.

Are you a litigator? It seems like it would fit my personality. My son's soccer coach is one of the most well respected (if there is such a thing) criminal defense attorneys in the city, and he said that the prosecutor route was the way to go. Hopefully there is going to be a scholarship involved, so paying student loans will be less of a burden and doing that will be a bit easier for me.

 

[CentralCaliBike]

Are you a litigator? It seems like it would fit my personality. My son's soccer coach is one of the most well respected (if there is such a thing) criminal defense attorneys in the city, and he said that the prosecutor route was the way to go. Hopefully there is going to be a scholarship involved, so paying student loans will be less of a burden and doing that will be a bit easier for me.

I have prosecuted a few cases - right now there is not much in the way of scholarship payments for government attorneys although there has been a bill in congress for a decade or more since the income of government attorneys is much lower than the private bar. There is a concern that most attorneys will spend a couple of years prosecuting and, once they get some experience, leave for the greener pastures of private practice.

 

[Anonymous]

I have prosecuted a few cases - right now there is not much in the way of scholarship payments for government attorneys although there has been a bill in congress for a decade or more since the income of government attorneys is much lower than the private bar. There is a concern that most attorneys will spend a couple of years prosecuting and, once they get some experience, leave for the greener pastures of private practice.

No, I meant scholarships from the Charlotte School of Law. But it would be nice to see the government do something like that because the pay is pretty low.

 

[CentralCaliBike]

No, I meant scholarships from the Charlotte School of Law. But it would be nice to see the government do something like that because the pay is pretty low.

I wish I knew but the closest I have been to NC is SC. I took a couple of classes at the National Advocacy Center at the University of South Carolina in Columbus. I really liked the environment there and took an afternoon trip to Charleston which was even better.

 

[Anonymous]

I wish I knew but the closest I have been to NC is SC. I took a couple of classes at the National Advocacy Center at the University of South Carolina in Columbus. I really liked the environment there and took an afternoon trip to Charleston which was even better.

Yea, my two best friends live in Charleston and we go pretty often. I love that city. I do get out to Cali too as my wife is from Nevada City and her parents still live there. I love it out there too.

 

[CentralCaliBike]

Yea, my two best friends live in Charleston and we go pretty often. I love that city. I do get out to Cali too as my wife is from Nevada City and her parents still live there. I love it out there too.

You should plan to come out in July next time and ride the Markleeville Death ride - the ride is not too far from Nevada City. Although, if you want a challenge, try the Climb to Kaiser in June (starts out of Clovis - which is basically Fresno).

Also, riding got me through law school and the bar exam - plan to scheduling regular riding along with your classes.

 

[RTMcFadden]

I also wondered about why he focused only on spiking the retentate. Surely one would need to spike the whole pot of p1ss, that the 20ml for filtering was taken from in the first place?....unless the whole pot is filtered, which would seem odd to me (especially as they asked for permission to split it and re-test after the results were out, so there must have been some p1ss left). If the idea is to run the test once and calculate the volume of rEPO required to spike the whole pot, the spiking volume should be big enough to be practical. But that calculation would depend on knowing what fraction of total isoforms survives the filtration process - do you have a view on whether this information might have been available? Regardless of whether it's doable or not, running tests on samples to calculate the volume needed to spike, in a way that would show expected trends in EPO doping, requires elaborate and deliberate sabotage.

Ok, I’ll try to answer your questions somewhat in order.

EPO Procedure Step 1
I’m not quite sure why your asking this question, but let me try to answer it. Let’s assume you had some chemistry in high school. There your would have probably learned about the concept of recrystallization. Basically you dissolve the substance, salt it out of solution, filter, dissolve, salt, filter, … Usually done by controlling temperature (steam bath – ice bath). Each cycle of the process yields a more pure sample (retentate). As a matter of fact, retentate simply means the stuff retained, or left on the filter. You simply continue cycling the process until the desired level of purity is achieve.

The same concept applies to the “retentate” in the EPO test. It’s just that instead of using solubility to differentiate the compound of interest from the contaminants, you’re using molecular weight. In addition, instead of using temperature, your using centrifugal force. Notwithstanding, based on the design of the experiment (DOE), the object would be the opposite. Instead of removing contaminants, you would be introducing one, specifically rEPO.

Spiking the retentate
Now, based on the above, it doesn’t really matter when you spike. You could spike an alloquat of urine, then purify, or you can purify, spike, and re-purify. What’s important here is that you first know the concentration of rEPO in the sample. You can determine this by purifying a different alloquat of sample, or working with the one you have. However, if you only have one sample (alloquat), then you would have to start with the retentate. What’s important here, based on the DOE, is that there is no “contaminant” (rEPO) in the original sample. Although, valid results can be obtained using samples already containing the “contaminant” it makes the entire process much more difficult.

Sample Size
According to Ashenden (http://nyvelocity.com/content/features/2009/spiking-armstrongs-99-samples) the original sample size was about 125ml. From that 20ml aliquots were taken, and I assume the rest was disgarded. One aliquot was tested in 99, the A Sample. The second aliquot (B Sample) was store for future reference. So, for this study, the whole sample would be 20 ml. I would guess that the reason they shoe 20ml was it provided enough sample for testing, on average, and it was a small enough to store efficiently.

Surviving Isoforms
This is the point of a recovery study. Filtration methods are normally prone to loss, which we’ll call mechanical loss. In addition, there are issues with protein binding, and degradation and other things. First, step is to identify if they is loss, then the second step is to identify where. If it’s mechanical loss, you look to modify the method. Note. that I believe this type of filtration is less prone to loss than you would see from a recrystallization process, but thinking is not the same as knowing.

In closing, let me apologize if you feel that I’m being pedantic or condescending, I don’t mean to be either. It’s just that I’m not sure how familiar you are with the subject matter, so I’m taking it slow. More to follow.

 

[CentralCaliBike]

In closing, let me apologize if you feel that I’m being pedantic or condescending, I don’t mean to be either. It’s just that I’m not sure how familiar you are with the subject matter, so I’m taking it slow. More to follow.

Glad to see someone else used the term pedantic in this thread

It appears that you are stating that it is clearly possible to spike the tests - I guess the question I would have is, even if possible, is it likely to have occurred in this case. Since I do not know the bias or character of the lab techs I really would have to think that the likelihood of spiking the test is unlikely (for those of you who think I am biased in favor of LA) but I also would not claim it is impossible to believe the tests were manipulated.

 

[RTMcFadden]

Yes but that would only mean that 3 to 5 times the volume would need to be added (to the retentate), and I suspect the volume would need to be 1000 times bigger to make the spiking straightforward. That's because Ashenden says "So if you wanted to add 0.0037674 UI you would add 0.000000113 ml (i.e., 0.6ml divided by 20,000/0.0037674 = 0.000000113 ml) of the synthetic EPO solution. This is 0.113 microlitres." But 0.000000113mL is 0.113 nano-litres (for the non-geeks that's 1000 times smaller, which really is tiny and it could well be difficult to spike accurately even with dilution techniques.) So maybe he is right in essence, and the source of the confusion is that he has just befuddled his units.

Dilution of Standard Solution for Spiking
From the information you provided Ashenden started with a 20,000 IU/ml concentration. In addition, he identified the target concentration as 0.0037674 UI, or 3.7674 mIU. I would start with the singe-dose preservative free vial, which has a concentration of 2,000 IU /ml. Then, using class A volumetric glassware, I would proceed as follows.

Aliquot Final volume Final Concentration
1 ml (2,000 IU /ml) 1000 ml 2 IU/ml
1 ml (2.0 IU /ml) 1000 ml 0.002 IU/ml = 2 mIU

Using the final standard solution identified above, I would probably pipet 2 ml of solution into a 10ml flask add my 20 ul of retentate, and mix. Then I would proceed with the sample prep with a known concentration of 4mIU. The volumes here could also be doubled, or modified to provide whatever concentration in this range that you want. Remember, that is a recovery study you’re usually targeting 100% of the expected unknown concentration, but you technically target additional concentrations.

 

[RTMcFadden]

Glad to see someone else used the term pedantic in this thread

It appears that you are stating that it is clearly possible to spike the tests - I guess the question I would have is, even if possible, is it likely to have occurred in this case. Since I do not know the bias or character of the lab techs I really would have to think that the likelihood of spiking the test is unlikely (for those of you who think I am biased in favor of LA) but I also would not claim it is impossible to believe the tests were manipulated.

Therein lies the rub. It equally as hard to prove that it did happen, as that it didn't. See, this could be done with out generating any type of paper trail. I could, with malice and forethought, do this without anyone being able to discover it. If I had people colluding with me, it wouldn’t even take forethought.

On the other hand, if I were conducting research, specifically performing a recovery study, I could do this with everyone’s full knowledge and without malice. Once the results are generated, they’re generated. If someone inadvertently misinterpreted these “research result” for actual same results, they could be passed along without malice. Yes, I know this scenario is a stretch, but I use it to demonstrate that it could happen without a massive conspiracy. The easiest way to disprove this scenario, would be to ask the right person, which isn’t me. Absent a response to the affirmative, I don’t believe this could have been done without malicious intent. And I don’t know the scientists so I can’t vouch for their character. Nevertheless, I choose to believe people, especially scientists, to be honorable.

 

[Race Radio]

Ok, I’ll try to answer your questions somewhat in order.

EPO Procedure Step 1
I’m not quite sure why your asking this question, but let me try to answer it. Let’s assume you had some chemistry in high school. There your would have probably learned about the concept of recrystallization. Basically you dissolve the substance, salt it out of solution, filter, dissolve, salt, filter, … Usually done by controlling temperature (steam bath – ice bath). Each cycle of the process yields a more pure sample (retentate). As a matter of fact, retentate simply means the stuff retained, or left on the filter. You simply continue cycling the process until the desired level of purity is achieve.

The same concept applies to the “retentate” in the EPO test. It’s just that instead of using solubility to differentiate the compound of interest from the contaminants, you’re using molecular weight. In addition, instead of using temperature, your using centrifugal force. Notwithstanding, based on the design of the experiment (DOE), the object would be the opposite. Instead of removing contaminants, you would be introducing one, specifically rEPO.

Spiking the retentate
Now, based on the above, it doesn’t really matter when you spike. You could spike an alloquat of urine, then purify, or you can purify, spike, and re-purify. What’s important here is that you first know the concentration of rEPO in the sample. You can determine this by purifying a different alloquat of sample, or working with the one you have. However, if you only have one sample (alloquat), then you would have to start with the retentate. What’s important here, based on the DOE, is that there is no “contaminant” (rEPO) in the original sample. Although, valid results can be obtained using samples already containing the “contaminant” it makes the entire process much more difficult.

Sample Size
According to Ashenden (http://nyvelocity.com/content/features/2009/spiking-armstrongs-99-samples) the original sample size was about 125ml. From that 20ml aliquots were taken, and I assume the rest was disgarded. One aliquot was tested in 99, the A Sample. The second aliquot (B Sample) was store for future reference. So, for this study, the whole sample would be 20 ml. I would guess that the reason they shoe 20ml was it provided enough sample for testing, on average, and it was a small enough to store efficiently.

Surviving Isoforms
This is the point of a recovery study. Filtration methods are normally prone to loss, which we’ll call mechanical loss. In addition, there are issues with protein binding, and degradation and other things. First, step is to identify if they is loss, then the second step is to identify where. If it’s mechanical loss, you look to modify the method. Note. that I believe this type of filtration is less prone to loss than you would see from a recrystallization process, but thinking is not the same as knowing.

In closing, let me apologize if you feel that I’m being pedantic or condescending, I don’t mean to be either. It’s just that I’m not sure how familiar you are with the subject matter, so I’m taking it slow. More to follow.

As none of what you have written makes sense I choose to believe Ashenden and Parisotto, both of whom have years of experience and helped develop the tests and procedures.

 

[Digger]

Therein lies the rub. It equally as hard to prove that it did happen, as that it didn't. See, this could be done with out generating any type of paper trail. I could, with malice and forethought, do this without anyone being able to discover it. If I had people colluding with me, it wouldn’t even take forethought.

On the other hand, if I were conducting research, specifically performing a recovery study, I could do this with everyone’s full knowledge and without malice. Once the results are generated, they’re generated. If someone inadvertently misinterpreted these “research result” for actual same results, they could be passed along without malice. Yes, I know this scenario is a stretch, but I use it to demonstrate that it could happen without a massive conspiracy. The easiest way to disprove this scenario, would be to ask the right person, which isn’t me. Absent a response to the affirmative, I don’t believe this could have been done without malicious intent. And I don’t know the scientists so I can’t vouch for their character. Nevertheless, I choose to believe people, especially scientists, to be honorable.

So, even if it is scientifically possible to spike the samples, do the odds of 480 to 1 mean anything to you?

 

[CentralCaliBike]

So, even if it is scientifically possible to spike the samples, do the odds of 480 to 1 mean anything to you?

I think you might have missed the point - unlikely that the samples were spiked - if they were, it was intentional - if intentional the odds really do not count.

 

[Sprocket01]

Someone else to throw into the mix. This is an article about possible false positives from EPO urine tests. Some said that it could not happen, but these guys reckon it can.

-------------------------
False-positive detection of recombinant human erythropoietin in urine following strenuous physical exercise

http://bloodjournal.hematologylibrary.org/cgi/content/full/107/12/4711

Monique Beullens, Joris R. Delanghe, and Mathieu Bollen
From the Division of Biochemistry, Department of Molecular Cell Biology, Faculty of Medicine, Catholic University of Leuven, Belgium; and the Department of Clinical Chemistry, University Hospital, Ghent, Belgium.

Erythropoietin (Epo) is a glycoprotein hormone that promotes the production of red blood cells. Recombinant human Epo (rhEpo) is illicitly used to improve performance in endurance sports. Doping in sports is discouraged by the screening of athletes for rhEPO in urine. The adopted test is based on a combination of isoelectric focusing and double immunoblotting, and distinguishes between endogenous and recombinant human Epo. We show here that this widely used test can occasionally lead to the false-positive detection of rhEpo (epoetin-) in postexercise, protein-rich urine, probably because the adopted monoclonal anti-Epo antibodies are not monospecific.

 

[Race Radio]

I think you might have missed the point - unlikely that the samples were spiked - if they were, it was intentional - if intentional the odds really do not count.

How did they figure out which samples were Armstrong's?

 

[RTMcFadden]

spike samples in a varying pattern[/B] that would match the fluctuations seen by EPO dosing approx every 4 days. Feel free to explain it slowly.

I'm not suggesting that the samples were spiked in a varying pattern. I'm suggesting that the results indicate that there may be some varaibility in the test method. That's why looking at the results as % is a little frustrating to me, as it could masks what may be going if this were a recovery study.

This is also what I find frustrating about Ashenden. He deliberately witholds some key information (data), which he specifically admitted to in haematologica/the hematology journal | 2006]. Now, I'm sure the conspiracy theorists will say that to hide the truth. Others (including Ashenden) will say it's for good reason. Either to keep the cheaters from gaining in sight into what they are really doing, or to protect subjects privacy, or something else. Everyone has to make up their own mind, it's just frustrating to me because it gets in the way of see the complete picture.

 

[RTMcFadden]

So, even if it is scientifically possible to spike the samples, do the odds of 480 to 1 mean anything to you?

No, 480 to 1 just makes me laugh.

 

[Race Radio]

Someone else to throw into the mix. This is an article about possible false positives from EPO urine tests. Some said that it could not happen, but these guys reckon it can.

-------------------------
False-positive detection of recombinant human erythropoietin in urine following strenuous physical exercise

http://bloodjournal.hematologylibrary.org/cgi/content/full/107/12/4711

Monique Beullens, Joris R. Delanghe, and Mathieu Bollen
From the Division of Biochemistry, Department of Molecular Cell Biology, Faculty of Medicine, Catholic University of Leuven, Belgium; and the Department of Clinical Chemistry, University Hospital, Ghent, Belgium.

Erythropoietin (Epo) is a glycoprotein hormone that promotes the production of red blood cells. Recombinant human Epo (rhEpo) is illicitly used to improve performance in endurance sports. Doping in sports is discouraged by the screening of athletes for rhEPO in urine. The adopted test is based on a combination of isoelectric focusing and double immunoblotting, and distinguishes between endogenous and recombinant human Epo. We show here that this widely used test can occasionally lead to the false-positive detection of rhEpo (epoetin-) in postexercise, protein-rich urine, probably because the adopted monoclonal anti-Epo antibodies are not monospecific.

You may want to post a study that has not been completely discredited.

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

the compound they are discussing is not rhEpo. If the compound detected can be identified as not rhEpo, then it cannot cause a false positive. This term sensationalizes an otherwise interesting case report that could in due course contribute to the body of science.

and

Beullens et al do not state the criteria they used to make the "false-positive" claim. Using the WADA criteria, the "false-positive" electropherogram1(Fig1A) is clearly negative.

 

[Race Radio]

No, 480 to 1 just makes me laugh.

I find that you cannot answer how they found out which samples were Armstrong's equally hilarious.

You have confused word count with content.

 

[RTMcFadden]

You may want to post a study that has not been completely discredited.
http://bloodjournal.hematologylibrary.org/cgi/content/full/108/5/1778

A article posted in a peer-reviewed journal and then discredited by a letter to the editor. Troubling indeed.

 

[CentralCaliBike]

How did they figure out which samples were Armstrong's?

As mentioned earlier - if it was intentional, figuring out the samples would be part of the malice. DNA could be used to determine who belonged to the sample (although finding DNA in urine is much more difficult than blood it is possible). Again, I doubt this scenario without evidence of a conspiracy, but I think you cannot claim it is impossible.

 

[RTMcFadden]

As mentioned earlier - if it was intentional figuring out the samples would be part of the malice. DNA could be used to determine who belonged to the sample (although finding DNA in urine is much more difficult than blood it is possible). Again, I doubt this scenario without evidence of a conspiracy, but I think you cannot claim it is impossible.

Boy, I never ventured down that path. Good catch.