AFLD offers to retest Lance's 1999 pee

On Oct 6, 11:37*am, MagillaGorilla wrote:
Michael Press wrote:
In article ,
*MagillaGorilla wrote:

Bob Schwartz wrote:

MagillaGorilla wrote:

When the French ladies at the LNDD who run the tests and determine the
positivity run an EPO test on sample #60152, how the hell do they know
it's Lance's to 'get him?'

Dumbass,

The claim was that they ****ed up the test by virtue of not knowing
how to run the equipment they were using. This is agreed upon by
everyone involved except you.

Bob Schwartz

That's like saying if you run an assembly line at the Ford Motor Plant
in Michigan improperly, you will get a Mercedes instead of a broken Ford.

You cannot get an EPO positive from a sample that was run improperly or
from a degraded sample. *You would simply get a negative result if
either occurred.

Also, there was no claim that the women ran the EPO tests improperly
since WADA never cooperated with the so-called independent UCI
investigation (a joke in and of itself considering the investigator was
hired by Verbruggen).

Can you show me any link that says they ran the EPO tests improperly?
No, you cannot. *So where do you come up with this stuff?

Can you show me the detailed protocols for detection of EPO,
and how it is impossible for the test to indicate EPO when
no EPO is present? Pretend that I do not consider the assembly
line analogy impossibly lame.

It's a complex protein. The test detects a complex protein through gel
electrophoresis. How can you get the EPO protein if it's not there?

That's almost like saying it's possible to find your DNA in water that
doesn't contain your DNA. *Not possible.

Magilla

dumbass,

one possibility is rutger beke proteins, the supposed mechanism is
here :

http://www.rutgerbeke.com/en/news/march082005.asp

but based on that it seems like it should be pretty easy to correct
for.

On Oct 6, 11:29*am, MagillaGorilla wrote:

The UCI is the union for the riders? *Absolutely not. *That would be
like saying the NBA is the union for basketball players. *In fact,
players need to form a player's union specifically to fight the NBA (and
teams).

dumbass,

the UCI is supposed to be the union for riders.

the UCI rulebook gives guidelines on what the teams are supposed to
pay riders and what organizers are supposed to provide in terms of
prizemoney and other facilities. that is something like a union.

the NBA on the other hand actually generates it's own revenue by
organizing the events and through it's franchisees contracts athletes.
the UCI doesn't do either of these things. the UCI generates revenue
by asking organizers and athletes/teams to hand over a chunk of their
earnings.

the reason the UCI does a poor job is because it is torn between
representing the interests of euro pro cycling and all the other
amateur/small-time disciplines and governing bodies it represents.

the NBA is a member of FIBA the int'l gov. body for basketball, which
makes it possible for NBA players to play in the olympics, but FIBA is
not involved in the dealings between the NBA and it's players (for
example FIBA doesn't intervene when the commissioner suspends ron
artest).

Amit Ghosh wrote:
On Oct 6, 11:29 am, MagillaGorilla wrote:


The UCI is the union for the riders? Absolutely not. That would be
like saying the NBA is the union for basketball players. In fact,
players need to form a player's union specifically to fight the NBA (and
teams).


dumbass,

the UCI is supposed to be the union for riders.

the UCI rulebook gives guidelines on what the teams are supposed to
pay riders and what organizers are supposed to provide in terms of
prizemoney and other facilities. that is something like a union.



The reason why this isn't true is because there is NO representation in
the UCI by riders. Therefore, by definition, the UCI is not their
union. They are simply a licensing and rules-making body that offers
some claimed protections to riders, although this is unclear since I can
find 20x as many rules that penalize a rider as do reward them.

The definition of a union is an organization that solely represents the
best interest of the riders against "management" - whether its teams or
the UCI.


Magilla

Donald Munro wrote:
Kyle Legate wrote:
How detailed do you want?

The EPO test uses a Western blot detection method. What this entails is
the separation of proteins according to size and recognition with a
specific antibody.

Interesting description of the process, thanks.

Thankfully the lemon club must have started their winter break, thus
allowing the president to make detailed contributions to rbr.

It's been a long season. After the Worlds I had nothing left.

Amit Ghosh wrote:

one possibility is rutger beke proteins, the supposed mechanism is
here :

http://www.rutgerbeke.com/en/news/march082005.asp

but based on that it seems like it should be pretty easy to correct
for.

It's most easy to correct for if your bull**** meter is properly
calibrated. His claims are outrageous! I mean, elevated urinary pH from
bacterial contamination?! Any microbiologist will tell you that the
presence of bacteria in urine would lower the pH.

Also, BLAST the EPO sequence against the entire protein database, and
alpha-ACT is nowhere in sight. It doesn't matter if it runs at the same
position in the gel if the antibody can't detect it.

As an addendum to my wordy earlier post. Rutger Beke's scientifically
incapable team refers to isoelectric focusing. This amounts to
separating proteins by charge rather than by charge/mass ratio. I stand
corrected. Endogenous and exogenous EPO are still separated from one
another.

On Oct 7, 10:06*am, MagillaGorilla wrote:
Amit Ghosh wrote:

The reason why this isn't true is because there is NO representation in
the UCI by riders. *

dumbass,

actually the gov. bodies all have a riders representative. i don't
know how UCI rules are formed, but supposedly there is supposed to be
an input from the membership (riders,officials,teams,organizers).

Therefore, by definition, the UCI is not their
union. *They are simply a licensing and rules-making body that offers
some claimed protections to riders, although this is unclear since I can
find 20x as many rules that penalize a rider as do reward them.

many of the rules are there to make the race fair. so one cheating
rider makes the race unfair to all the non-cheaters. so in theory
penalizing the occasional cheater benefits the majority of riders.
they apparently hadn't foreseen a scenario where 80% of the riders are
potentially cheaters.

The definition of a union is an organization that solely represents the
best interest of the riders against "management" - whether its teams or
the UCI.

riders are paid by their teams or by race organizers, so in essence
those two parties are the "management" and the UCI is just a body that
rubber stamps everything. as i've been saying it's a system suited for
managing amateur sports, but it doesn't work in a situation where some
athletes are being paid large sums of money to compete.

Amit Ghosh wrote:

On Oct 7, 10:06 am, MagillaGorilla wrote:

Amit Ghosh wrote:


The reason why this isn't true is because there is NO representation in
the UCI by riders.


dumbass,

actually the gov. bodies all have a riders representative. i don't
know how UCI rules are formed, but supposedly there is supposed to be
an input from the membership (riders,officials,teams,organizers).


The devil is in the details. Those "reps" have no power. They are
simply an advisory board. Therefore, they have no function whatsoever.

Second, the idea that former cyclists even know what the **** they're
doing on one of these boards is laughable given the bull**** that's
already in the UCI rulebook.

Magilla

Amit Ghosh wrote:

On Oct 7, 10:06 am, MagillaGorilla wrote:

Amit Ghosh wrote:


The reason why this isn't true is because there is NO representation in
the UCI by riders.


dumbass,

actually the gov. bodies all have a riders representative. i don't
know how UCI rules are formed, but supposedly there is supposed to be
an input from the membership (riders,officials,teams,organizers).


Therefore, by definition, the UCI is not their
union. They are simply a licensing and rules-making body that offers
some claimed protections to riders, although this is unclear since I can
find 20x as many rules that penalize a rider as do reward them.


many of the rules are there to make the race fair. so one cheating
rider makes the race unfair to all the non-cheaters. so in theory
penalizing the occasional cheater benefits the majority of riders.
they apparently hadn't foreseen a scenario where 80% of the riders are
potentially cheaters.



The problem with the UCI is they selectively enforce rules. For
example, the 3km rule was not enforced when Levi crashed outside the 3km
zone at AMGEN. You'll never see an NBA official count a shot as 3
points if it's inside the 3-point line just because he "thinks it's
unfair" that the home crowd favorite didn't shoot behind the line.

As for doping, the UCI is still ****ed up. How many people has the
biological passport system caught so far? NONE.

Schumacher was supposedly part of that - yet the CERA EPO he was taking
didn't affect his numbers? Obviously there's something wrong with this
system.

The other thing you have to ask is how come the AFLD caught all these
riders and the UCI didn't catch a single one? The Tour was held outside
the aegis of the UCI this year - how come the UCI didn't target riders
at the Tour?

The UCI catches like 2 rider/year - that tells me they are just
soft-pedaling this whole anti-doping thing.


Magilla

In article ,
MagillaGorilla wrote:

Michael Press wrote:

In article ,
MagillaGorilla wrote:


Bob Schwartz wrote:


MagillaGorilla wrote:


When the French ladies at the LNDD who run the tests and determine the
positivity run an EPO test on sample #60152, how the hell do they know
it's Lance's to 'get him?'


Dumbass,

The claim was that they ****ed up the test by virtue of not knowing
how to run the equipment they were using. This is agreed upon by
everyone involved except you.

Bob Schwartz


That's like saying if you run an assembly line at the Ford Motor Plant
in Michigan improperly, you will get a Mercedes instead of a broken Ford.

You cannot get an EPO positive from a sample that was run improperly or
from a degraded sample. You would simply get a negative result if
either occurred.

Also, there was no claim that the women ran the EPO tests improperly
since WADA never cooperated with the so-called independent UCI
investigation (a joke in and of itself considering the investigator was
hired by Verbruggen).

Can you show me any link that says they ran the EPO tests improperly?
No, you cannot. So where do you come up with this stuff?


Can you show me the detailed protocols for detection of EPO,
and how it is impossible for the test to indicate EPO when
no EPO is present? Pretend that I do not consider the assembly
line analogy impossibly lame.



It's a complex protein. The test detects a complex protein through gel
electrophoresis. How can you get the EPO protein if it's not there?

Okay. I will trust you on this.

That's almost like saying it's possible to find your DNA in water that
doesn't contain your DNA. Not possible.

Another cool analogy.

--
Michael Press

In article ,
Kyle Legate wrote:

Michael Press wrote:


Can you show me the detailed protocols for detection of EPO,
and how it is impossible for the test to indicate EPO when
no EPO is present? Pretend that I do not consider the assembly
line analogy impossibly lame.

How detailed do you want?

The EPO test uses a Western blot detection method. What this entails is
the separation of proteins according to size and recognition with a
specific antibody.

Proteins from a riders urine (or blood also, these days) are separated
by SDS-PAGE: this is a slab of porous polyacrylamide with holes of a
defined size that let small proteins sail through quicker than large
proteins. The proteins are driven through using voltage (or current,
depending on the specific PAGE system). Prior to loading the sample on
the gel the proteins are boiled with a detergent which serves the dual
purpose of denaturing them and applying a constant charge per unit
length so indeed small proteins run faster than large ones. Once
separation is complete, the proteins are transferred from the
polyacrylamide slab to a support membrane, again through the application
of current, for the remaining steps.

Specific proteins are detected through the use of antibodies. As you
know, the antibodies in your body are highly specific against certain
pathogens; commercial antibodies are likewise highly specific. For
detection of linear protein samples on a gel an antibody is raised in an
animal (usually a rabbit or mouse) against a synthetic peptide (the
antigen) corresponding to the protein of interest. Prior to making the
antibody the sequence of the antigen is designed and checked using an
online tool called BLAST. BLAST aligns your input sequence against all
known protein, peptide, and theoretical gene product sequences from all
organisms in the database (hundreds) to determine how specific for your
protein your proposed antigen is. Once the peptide sequence BLASTS only
for EPO, it is synthesized, injected into the animal, and in about 2
months you have the antibody. To separate the antibody of interest from
the antibodies already present in the animal, there are two approaches.
Either the antibody is pulled out of the serum using the original
peptide as bait, or a cell line is derived which makes only the antibody
of interest (significantly more work but more desirable for long term
production of antibody). I hope this convinces you that the antibody
used in the EPO test specifically recognizes only EPO and nothing else,
no bacterial protein, no peptide fragment from a different protein in
the urine, only EPO.

Check.

The support membrane containing the size-separated proteins is incubated
with the antibody against EPO. This antibody is then recognized by
another antibody (anti-rabbit or anti-mouse, usually raised in goat or
donkey), coupled to an enzyme that allows for detection via fluorescent
breakdown products from the developing solution. To wit, the EPO-primary
antibody-secondary antibody complex will glow when a solution of
hydrogen peroxide and Luminol is placed on the membrane. A picture is
taken with film specific for the purpose, or with a CCD camera in a dark
box. Based upon the position of the glowing band on the membrane, even
your mother would be able to tell if Armstrong took EPO. Here's why.

Your kidney cells produce endogenous EPO. Part of the production process
is the addition of sugars to the protein, which makes it migrate as a
heavier protein in the polyacrylamide gel. Synthetic EPO is also made by
cells, but these cells are from chinese hamster ovary. The different
species and organ of origin means that these cells have different sugar
attachment enzymes, which leaves the EPO differently labeled when it is
made from these cells. Your body's EPO and synthetic EPO are therefore
slightly different in mass and will run to different positions in the
gel. What a negative test will look like is a nice, clean, single band
owing to the athlete's own EPO. What a positive test will look like is
this same band, but the addition of a second band nearby. An
inconclusive test would look like either nothing at all, or a smear
beginning at the normal position of EPO and extending down the gel until
the antibody recognition site is destroyed. I presume that years-old
urine samples were being tested to determine whether after all these
years EPO would now present as a smear. Since they found exogenous EPO
in Armstrong's samples I presume that the integrity of the EPO was just
fine, and they saw two distinct bands.

Based on the specificity of the antibody, outlined above, and the
various things that can go wrong with the test, it it only possible to
have false negatives based on inconclusive test results, and not false
positives. In all my years of doing Western blots (for research
purposes) I have encountered false negatives and inconclusive results
many, many times, but I have never encountered a false positive. It is
just not in the nature of the method to give false positives.

Okay, thanks for the write up.

But how does EPO get into the urine?
Protein in the urine is a bad thing, yes?
Indicates serious kidney problem.
Is it that tiny amounts in the urine
are not a problem?

--
Michael Press