Hearing Monday - Shackleton Testimony Part I

[Note -- streaming has corrected passwords]

Cedric Shackleton, of Children's Hospital Oakland, is testifying for USADA. He has 240 or so published papers, and has never testified in a doping case before. From his bio page:

Dr. Cedric Shackleton is a Senior Scientist at CHORI as well as being a Professor (Honorary) at the Department of Medical Sciences, University of Birmingham, England. He has worked at CHORI for over 20 years.

Since 1967, Dr. Shackleton has been an innovator in the use of mass spectrometetry for biochemical analysis. For most of this period his new studies have involved steroid metabolism, but in 1991 he was a pioneer in the use of new technique electrospray mass spectrometry for protein analysis. For several years his group acted as a resource within the Sickle Cell Center for characterization of variant hemoglobins. The central theme of his studies has been the utilization of the technique for clinical analysis.

He starts explaining testosterone metabolism, and carbon content. 1.1% of carbon on Earth is C13, with an extra neutron; the rest is C12. Organic entities - animals and plans have different values of each, based on diet; plants can be selective about what they extract from atmospheric C02.

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Pharmacutical testosterone is made from either yams or soy, and both have way more C13 than animals. The IRMS tests sees how much C13 is in the metabolites of testosterone in sample in question, and if it's out of whack with respect to what is normal for the person, you can conclude some of the testosterone is pharmaceutical.

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Young leads him into metabolic breakdown of testosterone for different administration methods, and he's explaining that some methods yiled more of one product than another, at different rates. This is probably attempting to lead to the UCLA/Cologne studies that are likely to be rejected as unpublished/unreviewed.

Q: Can one delta change indicate exogenous use?

A: Yes, but details are specific to individuals. You'd need a lot of experimentation to know exactly what is going on.

Q: Would individual variance account for -5 or -6 differences.

A: That's impossible.

Q: On exhibit 107, assuming all these results were from the same athlete, do you think these are inconsistent with steroid metabolism?

A: They could be totally consistent. Let me draw a picture.

Floyd is shown taking some gum out, and wrapping it. Someone in the press room suggests he sell it on ebay.

The board he tries to draw on falls over. Landis is smiling in disbelief at the show.

Shackleton's graph, artist's impression

A: There are no differences that can't be explained by normal metabolism over time.

Q: Could this be done by an athlete switching back and forth between delivery mechanisms?

A: Yes, but you'd have to experiment a lot; it's not predictable.

Young is trying to prepare a theory that Landis was switching back and forth between injections and gels and patches to explain the variances.

Q: What could you eat to produce a --6?

A: It's not possible.

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CROSS EXAMINATION

Jacobs is doing cross.

Q: Are you a medical doctor?

A: No.

Q: The metabolisation is always down, or might you see them going up?

A: One might kick in before the other.

Q: [something] would track exactly the same, right?

A: yes.

Q: With administration of topical (on skin), you said 5a will be reduced, but not 5b; what about when it gets to the liver.

A: Needs another step; skin only gets to step 1 on my chart; needs another to get to 5 andrsostinediol (sp?)

Q: Table 3 is table of values on a certain date. We should expect them to track same direction on pairs of metabolites. Didn't see the table to know what it was -- could be results of all tour IRMS tests.

A: yes, i think so.

Q: On the 13th, you see a jump; on the next it's way low. In that one day Jul 13 to 14 do you think the exogenous completely cleared the system

A: I'd have to defer to somebody who has studied this...

Q: So you haven't studied the time values of metabolizations as seen by IRMS?

A: No, my knowledge is conceptual, not current. I have not personally done that to know.

Q: Studies on T/E

A: Completely out of my area of expertise. I'm smart, and I've been doing GCMS for 40 years, but not specifically.

Q: Jul 18-20; One delta increases, one decreases. Make sense?

A: Yes.

Q: Contradicts earlier chart?

A: No, we don't know doses, intervals. Too many things we don't know.

Q: If we charted Jul 18 and Jul 20 on your chart, one would go up, and one down.

A: Not sure I understand. Fumbling, thinking. Since we don't know if there was a dose, or what it was.

Q: Please then draw us what appears to have happened from his data.

[objection: manually plotting is not his area of expertise]

Q: The graph you drew does not match the longitudinal, does it?

A: [pause] I'd still disagree with you on the basis, that without details and timings I can't say.

Q: We're going to draw out what we see, and you'll tell us if it's consistent with what you see of the longitudinal data, from Jul 18 to Jul 20.

[ objection about chart drawn by Paul Scott, because the points aren't at the calibration ]

Jacobs Chart, artists rendering

A: Dependent on a single sample at a given time. This assumes a single dose was taken. If there was a secondary does, it could go the other direction.

Q: So you are testifying they can go different directions?

A: Perhaps if there were multiple doses.

Q: If the pair is returning to normal, they are both going down, right?

A: Yes.

Q: At no point you are going to have one going up and one going down, are you?

A: I'm out of my league.

Q: So your chart is inaccurate, correct?

A: It's meant as an illustration.

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SUH doing Cross:

Q: Are you being paid?

A: $200/hr

Q: Total so far?

A: About $1800.

Q: Any other benefit?

A: No.

Q: Not an endrologist, not an IRMS expert?

A: Correct, but I work with IRMS experts.

Q: Are you an IRMS expert or not?

A: It's not that simple. There are people

Q: Not a GCMS expert.

A: I'm pretty good. I think so. 40

Q: All your testimony assumes the values are correct, right?

A: I reviewed the data.

Q: You assumed they were correct.

A: Yes.

Q: Did you look at the documents underlying the values?

A: Looked at them.

Q: Would you agree that good chromatography is essential to good IRMS results.

A: Essential.

Q: Are you familiar with this?

A: Yes, it's my paper.

Panel: is it marked?

Suh: It is now.

[exhibit 40 at USADA 1241.]

Q: Highlighted part.

[Copy of the document from witness. Young suggests cherry picked selection, give him a chance to refamiliarize himself. ]

[exhibit at 753]

Q: What does good chromatography mean to you?

[gets paper]

A: Optimum would be baseline separation with very high resolution columns. There are times where baseline separation is not possible. You could use it anyway.

Q: When would you use it?

A: I would be looking for an example in things that have already come up today, for instance, the Epi-T measurement.

Q: In what circumstances would you use a chromatogram without good separation?

A: Very important.

Q: You would never use one with good separation.

A: Yes.

Q: When would you use one with co-eluted peaks?

A: Difficult to answer; my line of work is different than this. In IRMS is very important to have peaks that are baseline separated. Might be a small shoulder the software can resolve. It's a dream that every component can be well separated. You get as close as you can to find what's important in a particular case.

Q: If you had software that would remove effects of co-elution, would you use it?

A: No, you shouldn't use it?

Q: Do you know what a sloping baseline is?

A: Yes.

Q: When would you use one?

A: It wouldn't bother me.

Q: Causes to downward and upwards sloping baseline?

A: Down is involatile materials in sample.

Q: Wouldn't know the CIR of those would you?

A: You could measure them.

Q: Hold that thought.

Q: This is USADA 173.

[ Young wants him to look at the real document, not the screen ]

Q: Does this show good separation.

A: Looks pretty good.

Q: What about the humps? The peaks show good separation?

A: Yes.

Q: USADA 349 Looking at the peaks, would you call those good?

A: Acceptable, not good.

Q: What do you mean acceptable?

A: I'd be quite happy with that.

Q: USADA 161

[ Suh changes tack ]

Q: USADA 171 I'd like to show you this chromatogram. You said those were good. Here's the GC chromatogram of the same sample. This is the same as the one you said was good.

Q: Why are there two chromatograms for each sample?

A: I'd rather someone else said.

Q: Where did this hump go? You don't know the C13 of that do you?

A: My argument would be that if that hump if it's 1-2% of the peak, it wouldn't be measurable even if it was included. It's irrelevant.

Q: You don't know where it is?

A: Don't know the C13 of it, do you?

Q: You can ask the person who did the analysis. It could have no affect on the result.

[ Testy exchange between between Young/Suh. Young thinks Suh is interrrupting the answer, and Suh says the witness is not answering the questions being asked. ]

Q: They should be proud of that chromatogram. I would be. And I was the first to do capillary chromatography of steroids.

Q: LNDD 991. No USADA number. See where I circled?

A: Yes.

Q: See where this has a downward sloping baseline.

A: Yes.

Q: Agree there is no peak separation here?

A: I think it's very good.

Q: They are touching each other!

A: I think it's rather good. If there's overlap it looks at about the 2% level, by eyeball.

Q: Do you know what the C12/C13 is of the eyeball.

A: I don't think you've done the math. If there was a component shared at 1/2% and it was at a really low value, say -35, it can't affect the ratios of adjacent large peaks. I doubt you'd even see a difference.

Q: Let's look at LNDD 894.

A: Not as nice, relatively good separation.

Q: How do you know the volume here isn't absorbed by this peak with this high background?

A: I don't think there can be enough material. Background is known for these runs, right? The CIR is known, isn't it? It's not my run.

Q: Not my run either.

A: However low the value, to markedly change the CIR form a -25 to a -29, you need one of equal or half again to affect the result. A very small component can not have that affect.

Q: But this is a bad chromatogram.

A: Obviously not as good.

Q: Can you prove to me that this matrix interference did not affect the AAF?

A: You're going to ask someone else.

Q: You can't prove it.

A: How can I be asked to prove it when it's not my data and its not my method. I wasn't there.

Q: Yes or no. Can you prove it doesn't affect the aaf.

[objection asked and answered ]

A: No I can't prove it. It's not my position.

Q: Shall we take a break for lunch?

RECESS