Doesn't Russian Doping Scandal Prove Canova is Right about EPO?

Update: the link to Oxygen is wrong. Searching for oxygen-hemoglobin dissociation curve in Wikipekia will give the correct page.

This page has some of the same information:

https://en.wikipedia.org/wiki/Oxygen_saturation_

(medicine)

1. Yes, I stated that malaria is a virus, but this because, without disctionary, I didn't know how to call the disease. Excuse me, but "protozoan parasite" is not part of my reduced English lexicon.

2. Of course all endurance training, included endurance training at low level, increases the total blood volume. The difference is that at sea level the increase is about 5%, while in altitude, for the best athletes when are in full training (a type of training for the best performances in the World), the percentage of increase can reach 25%, and it's not exactly the same thing...

3. In order to make your physiological vision more wide, go to read some scientific work from researchers who tried to analyze the problem. Frankly, I don't understand anything of what they wrote, eccept the part where they talk about "human mutant with increased affinity between Hb and O2".

Respir Physiol Neurobiol. 2007 Sep 30;158(2-3):121-7. Epub 2007 Mar 24.

The role of hemoglobin oxygen affinity in oxygen transport at high altitude.

Winslow RM1.

Author information

Abstract

Hemoglobin is involved in the regulation of O(2) transport in two ways: a long-term adjustment in red cell mass is mediated by erythropoietin (EPO), a response to renal oxgyenation. Short-term, rapid-response adjustments are mediated by ventilation, cardiac output, hemoglobin oxygen affinity (P50), barriers to O(2) diffusion, and the control of local microvascular tissue perfusion. The distribution of O(2) between dissolved (PO2) and hemoglobin-bound (saturation) is the familiar oxygen equilibrium curve, whose position is noted as P50. Human hemoglobin is not genetically adapted for function at high altitude. However, more specialized species native to high altitudes (guinea pig and bar-headed goose, for example) seem to have a lower P50 than their sea level counterparts, an adaptation that presumably promotes O(2) uptake from a hypoxic environment.

"HUMANS, NATIVE TO VERY HIGH ALTITUDE EITHER IN THE ANDES OR HIMALAYAN MOUNTAINS, ALSO CAN INCREASE O(2) AFFINITY, NOT BECAUSE OF A FUNDAMENTAL DIFFERENCE IN HEMOGLOBIN STRUCTURE OR FUNCTION, BUT BECAUSE OF EXTREME HYPERVENTILATION AND ALKALOSIS."

High Altitude Medicine & Biology

High Alt Med Biol. 2008 Jun; 9(2): 148–157.

doi: 10.1089/ham.2007.1079

PMCID: PMC3140315

Mechanisms of Hemoglobin Adaptation to High Altitude Hypoxia

Jay F. Storz 1 and Hideaki Moriyama2

Hemoglobin Function

Homotopic effects: cooperative O2 binding

The binding of O2 at each of the four heme irons in the Hb tetramer exhibits a positive cooperativity, meaning that O2 binding at one site increases the O2 binding affinity at each remaining site. Likewise, O2 unloading at one site decreases the O2 binding affinity at the remaining sites. This cooperativity among the four globin subunits of each Hb molecule enhances the efficiency of O2 loading and unloading for a given difference in pulmonary–tissue O2 tensions and is manifest in the sigmoid shape of the O2 equilibrium curve (Fig. 1). The cooperativity of O2 binding results from the fact that the binding of O2 to the heme iron of a given subunit produces a localized change in tertiary structure that is transmitted to adjacent subunits, thereby triggering the shift in quaternary structure (Perutz, 1970, 1979; Arnone, 1974; Baldwin and Chothia, 1979; Gelin et al., 1983; Perutz et al., 1987; Liddington et al., 1988). This oxygenation-linked shift in quaternary structure between the T- and R-states is central to the allosteric function of Hb as an O2 transport molecule.

Heterotropic effects: binding of allosteric effectors

The respiratory functions of Hb are a product of its intrinsic O2-binding affinity and its interactions with allosteric effectors such as protons, chloride ions, CO2, and organic phosphates. These effectors exert their influence on Hb–O2 affinity by binding more strongly to deoxy Hb, mainly at sites located at the N- and C-termini, thereby stabilizing the low-affinity T structure through the formation of additional salt bridges within and between subunits (Perutz, 1970, 1989; Bettati et al., 1983).

The allosteric effect of proton binding accounts for the typical reduction in Hb–O2 affinity at low pH (the Bohr effect) and facilitates O2 unloading under conditions of metabolic acidosis in working muscles. At physiological pH, the Bohr effect of human Hb is primarily attributable to proton binding at the following residues: α1(NA1)Val, α122(H5)His, β2(NA2)His, β82(EF6)Lys, β143(H21)His, and β146(HC3)His (Perutz et al., 1969; Kilmartin et al., 1978; Ho and Russu, 1987; Lukin and Ho, 2004). Chloride ions bind to one α-chain site between α1(NA1)Val and α131(H14)Ser and one β-chain site between β1(NA1)Val and β82(EF6)Lys (Riggs, 1988). Similarly, CO2 combines with the N-terminal residues of each subunit chain of deoxyHb (Arnone, 1974; Perutz, 1983). It has also been hypothesized that Cl− may modulate O2 affinity through delocalized electrostatic effects that do not involve binding at specific residues (Perutz et al., 1994). According to this view, Cl− partially neutralizes the excess of positive charges between the β-chains of deoxy Hb, thereby stabilizing the T-state conformation. IF THIS VIEW IS CORRECT, THEN IT IS POSSIBLE THAT HB-O2 AFFINITY COULD BE INCREASED NOT JUST BY SUBSTITUTIONS AT SPECIFIC CL-BINDING SITES, BUT ALSO BY SUBSTITUTIONS AT A NUMBER OF RESIDUE POSITIONS THAT INCREASE THE NET ELECTROPOSITIVITY OF THE CENTRAL CAVITY. 2,3-DPG carries four negative charges, which allows it to bind between the β-chains of deoxyHb by charge–charge interactions with the β1(NA1)Val residue of one chain and with β2(NA2)His, β82(EF6)Lys, and β143(H21)His of both chains (Fig. 4). In principle, substitutions at any one of these binding sites can alter the sensitivity of Hb to the various allosteric effectors, thereby altering the equilibrium between the T- and R-state quaternary structures. Since the binding of allosteric effectors typically stabilizes T-state deoxyHb, substitutions that inhibit effector binding will typically increase Hb–O2 affinity by shifting the equilibrium in favor of R-state oxyHb.

Pleiotropic Constraints on Hemoglobin Evolution

In addition to comparative studies of Hb function in animal species that are native to high and low altitude environments, the wealth of functional information about HUMAN HB MUTANTS ALSO PROVIDES INSIGHT INTO MOLECULAR MECHANISM THAT UNDERLIE CHANGES IN INTRINSIC HB-O2 AFFINITY AND SENSITIVITY TO ALLOSTERIC EFFECTORS (Dickerson and Geis, 1983). Some of the same mutations that alter Hb–O2 affinity of human Hbs may be involved in adaptive modifications of Hb function in other species. In humans, high-affinity Hb mutants are typically associated with reduced levels of tissue oxygenation, which results in polycythemia due to the increased production of erythropoietin. However, affinity-enhancing mutations that have deleterious effects in humans living under normoxic conditions may be physiologically advantageous in animal species that inhabit hypoxic environments where the preservation of arterial O2saturation is at a premium.

FUNCTIONAL INFORMATION ABOUT HUMAN HB MUTATIONS CAN ALSO PROVIDE INSIGHTS INTO THE NATURE OF PLEIOTROPIC CONSTRAINTS ON HB EVOLUTION (where pleiotropy here refers to the case where a single mutation has multiple effects on different aspects of protein structure or function). Information on pleiotropic constraints is important because it can provide clues as to why certain evolutionary pathways are followed more often than others. FOR EXAMPLE, EVEN IF MUTATION X AND MUTATION Y PRODUCE IDENTICAL EFFECTS ON ONE PARTICULAR ASPECT OF HB FUNCTION, SUCH AS O2-BINDING AFFINITY, the two mutations may have different pleiotropic effects on other aspects of Hb structure or function. THUS, tHE TWO MUTATIONS MAY DIFFER WITH RESPECT TO THEIR NET EFFECTS ON PHYSIOLOGICAL PERFORMANCE (AND FITNESS).

THE MAJORITY OF HUMAN HB MUTANTS THAT INCREASE O2-BINDING AFFINITY do so by destabilizing the T-state (Dickerson and Geis, 1983). This is typically accomplished by disrupting hydrogen bonds or salt bridges at the chain termini of deoxyHb. For example, three different α-chain mutations (Hbs Tarrant, Suresnes, and Legnano) favor the T-to-R transition by disrupting an important salt bridge between 126(H9)Asp and the C-terminal 141(HC3)Arg in deoxyHb. This is accomplished by eliminating a side chain at either one of the two residue positions.

THE AVAILABLE DATA ON HUMAN GB MUTANTS INDICATE THAT THERE ARE MANY POTENTIAL MUTATIONAL CHANGES THAT CAN PRODUCE AN INCREASED HB-O2 AFFINITY.

Convergent Evolution of Hemoglobin Function in High-Altitude Andean

Waterfowl Involves Limited Parallelism at the Molecular Sequence Level

• Chandrasekhar Natarajan,

• Joana Projecto-Garcia,

• Hideaki Moriyama,

• Roy E. Weber,

• Violeta Muñoz-Fuentes,

• Andy J. Green,

• Cecilia Kopuchian,

• Pablo L. Tubaro,

• Luis Alza,

• Mariana Bulgarella,

• Matthew M. Smith,

• Robert E. Wilson,

• Angela Fago,

• Kevin G. McCracken,

• Jay F. Storz

• Published: December 4, 2015

•

http://dx.doi.org/10.1371/journal.pgen.1005681

Parallel replacements that contribute to convergence in Hb function.

Parallel β94Asp→Glu replacements that occurred in high-altitude crested ducks and Puna teal are associated with pronounced increases in the O2-affinities of both HbA and HbD relative to variants of the same isoforms that predominate in low-altitude sister taxa (S9 and S10 Figs; S2 Table), although there is potentially confounding amino acid variation in the αA-globin gene (Fig 1). In the HbA isoforms of both species, β94Asp→Glu is associated with a slightly higher intrinsic O2-affinity (S9A and S9B Fig, S10A and S10B Fig), and in both HbA and HbD it is generally associated with a suppressed anion sensitivity (S9C and S9D Fig, S10C and S10D Fig). With regard to overall effects on Hb-O2 affinity, the β94Asp→Glu mutation appears to have similar affinity-enhancing effects on different genetic backgrounds, as revealed by comparisons between the same isoforms in different species (orthologous comparisons), and comparisons between different isoforms in the same and in different species (paralogous comparisons). The high- and low-altitude HbA variants of crested ducks exhibited a 1.5-fold difference in P50(KCl+IHP) (25.14 vs. 37.98 torr, respectively), as did the HbA isoforms of Puna teal and silver teal (27.32 vs. 39.66 torr, respectively). The fact that the differences in P50 were identical in magnitude in both pairwise comparisons suggests that the shared β94Asp→Glu replacement in high-altitude crested ducks and Puna teal accounts for all or most of the observed difference in HbA O2-affinity, and that the additional αA5Ala→Thr replacement in high-altitude crested ducks is less consequential. Moreover, the comparison between HbD variants of crested ducks cleanly isolates the effect of the β94Asp→Glu mutation, as there is no confounding variation in the αD-globin gene (Fig 1). This single amino acid replacement produced a 1.9-fold reduction in P50(KCl+IHP) (20.35 vs. 10.51 torr; S2 Table). The β94Asp→Glu mutation is therefore associated with significant increases in Hb-O2 affinity on all backgrounds in which it occurs.

IN HUMAN HB, MUTATIONAL REPLACEMENTS OF β94Asp INCREASE O2-AFFINITY AND GREATLY DIMINISH THE pH-DEPENDENCE OF O2-BINDING (Bohr effect) by disrupting the salt bridge between the carboxyl group of β94Asp and the imidazole group of the C-terminal β146His of the same β subunit [48–50]. Elimination of this intra-chain salt bridge destabilizes the low-affinity T-state conformation of the Hb tetramer, thereby increasing Hb-O2 affinity by shifting the allosteric equilibrium in favor of the high-affinity R-state. Disruption of the electrostatic interaction between β94 and β146 greatly diminishes the Bohr effect by attenuating the charge stabilization of β146His [51]. In human Hb, the β94Asp→Glu mutation diminishes the Bohr effect by ~50%, and crystallographic analysis confirmed that the salt bridge is not formed between β94Glu and β146His [50].

OK, for convergent evolution, the second fastest female marathoner , Liliya Shobukhova (Russia) - 2:18:20, was born near the Ural Mountains in Russia.

She was placed in a Sports Academy during part of her childhood, and developed her aerobic system as part of the curriculum. She would have received whatever chemicals the Soviets liked to use.

This does not explain Paula.

Your studies discuss the same oxygen-hemoglobin saturation curve and the effect of increasing pH by hyperventilation and respiratory alkalosis, which increases the binding affinity of Hb for oxygen.

Look at this part:

" In humans, high-affinity Hb mutants are typically associated with reduced levels of tissue oxygenation, which results in polycythemia due to the increased production of erythropoietin. However, affinity-enhancing mutations that have deleterious effects in humans living under normoxic conditions may be physiologically advantageous in animal species that inhabit hypoxic environments where the preservation of arterial O2saturation is at a premium."

The high affinity human hemoglobin mutants DECREASE tissue oxygenation, and the body adapts by increasing endogenous EPO production which increases the number of RBCs. These mutants are not seen frequently in Kenyan or Ethiopian runners living at high altitude. Mutant high-affinity hemoglobin may be an advantage is some animal species that inhabit hypoxic environments. Your last study is about high-altitude Andean crested ducks and Puna teal…which are high-altitude waterfowl.

I do not think that a study on hemoglobin evolution in crested ducks living at high altitude in the Andes mountains is relevant to Kenyan and Ethiopian runners living and training at high altitude in Kenya or Ethiopia.

Ridiculo.

Renato.

That article about hemoglobin evolution in the crested duck living (and training) at high altitude in the Andes mountains was really boring.

Here is the high altitude/high volume/high intensity training program that I will be doing in 2017:

https://drive.google.com/file/d/0B_zzkn1-wR0dVU5wX1hveWppeUE/view

If I do not run really well in 2017…then you are going to be in really big trouble.

OK?

rjm33 wrote:

Renato.

That article about hemoglobin evolution in the crested duck living (and training) at high altitude in the Andes mountains was really boring.

Here is the high altitude/high volume/high intensity training program that I will be doing in 2017:

https://drive.google.com/file/d/0B_zzkn1-wR0dVU5wX1hveWppeUE/view

If I do not run really well in 2017…then you are going to be in really big trouble.

OK?

What the he'll was this guy taking? 14 x 3min WTF.

Probably not. No one ever said how much 2:08 Kenyans would improve, if at all.

casual obsever wrote:

Do I understand that correctly, Renato: if a 2:04 Kenian runner takes EPO, he runs 2:05, and if a 2:08 Kenian takes EPO, he runs 2:06 - 2:07 (both with the full altitude set)?

How does that translate to Americans, who don't live at altitude? Would a 2:12 runner go to 2:10? And the 2:08 guy 2:07?

You always start from a wrong point : that the best in the World and good athletes of international level, but not top elite, can have the same type of attitude.

Try to look at the WR holder and athletes around their times in different perspective.

WHY ARE THEY THE BEST IN THE WORLD ? WHICH IS THE DIFFERENCE IN THEIR TALENT, ALLOWING THEM TO DO WHAT THE MOST PART OF STRONG ATHLETES ARE NOT ABLE TO DO ?

The theme is complicated, AND NEVER WAS STUDIED with specific subjects.

But is a fact that, for example, if we have two Kenyan athletes running 28' in 10 km with a training of normal level (not very hard and without geat intensity), and both come to me for having more serious training, in 4 months one is able to run 26'30" like Nicholas Kemboi, and another, training always together with same volume and same intensity, can only lower his PB to 27'30".

It's a fact that very few athletes are able to grow in very short time from 70% of their PB till the full shape, while I never met some European with the same attitude, and also the most part of African are not in this category.

About the studies I previously indicated, we can take two important informations : there are human mutants living in high altitude having the affinity between Hb and O2 higher than in other human beings, and the adaptation of their body (because the lack of O2 to their tissues) is compensated by an increased production of endogeneous EPO.

Is this one of the reasons ? I don't know, but what I know is that there is a "subclass" of top athletes having a different behavior from other strong runners. And to this subclass, ALL THE BEST IN THE WORLD BELONG.

All the studies about change of affinity Hb-O2, increase of RBC, differences in the respiratory system between sea level and altitude, REGARD PEOPLE MOVING FROM A NORMAL SITUATION OF SEA LEVEL TO AN ABNORMAL SITUATION OF ALTITUDE.

This means that all these studies speak about the REACTION OF THE BODY to a new situation, while the stable situation of Kenyans and Ethiopians must be specifically studied under the point of view of ADAPTATION.

The two situations are totally different, and the physiological answers are, in some case, at the opposite.

I'm not able to know in right way ALL WHAT HAPPENS in the body of top athletes born, living and training in altitude, but is of course a mistake to put them in the same pot with weaker athletes living and training at sea level for the most part of their seasons.

There is a reason if I ask that some scientist can start to study THE EFFECTS OF TRAINING of the best in the World, before speaking about doping. The fact that ALL THE BEST RECORD HOLDERS were able to run completely clean (some of them is/was my athlete, so I well know), and in spite of their records I was not totally happy about their performances (so, some WR is still far from the personal possibilities of the holder), makes me thinking that there is some physiological mechanism that THEY own, and other athletes with the same morphology and the same motivation don't have.

Again, are we able to answer to the question : WHAT IS TALENT FOR THE ENDURANCE EVENTS ? Because it's something we don't see, and never was studied by any scientist in the past and in the present.

Of course. If "you" are making the argument about anything, there is a burden of proof on "you". For example, equally "if you are making the argument that blood doping *DOES* work for a specific subclass, then the burden of proof shifts (back) to you." Existing studies on different populations do not relieve you of this burden, nor permit you to shift the burden onto others to prove the contrary. Both arguments remain unproven, requiring more data.Google "dangers of extrapolation" to see good reasons why observations of the 11:00 3K runner may fail to accurately predict what happens at the extremes.There is no Olympic event determining who has the highest RBC or delivers the most O2 to the muscles. The argument is not that physiological and biological mechanisms and body responses are vastly different, but how these responses impact performance improvements in running, as measured by time over a distance.As we have just seen (and we've seen this before), there are studies that show various high altitude inhabitants have adapted in different ways to low oxygen environments. This is enough to suggest that extra RBCs from EPO may be superfluous in populations that are not starved for oxygen under the increased demands of competition. This is not a proof, but it reinforces the need to validate the assumptions between the two groups with known differences, in order to build an accurate performance model.

Mr. Obvious wrote:

... if you are making the argument that blood doping does not work for a specific subclass, then the burden of proof shifts to you. It is the most reasonable assumption that the same mechanisms that work in study populations also work in elite well trained athletes.

I just don't see good reason to believe that athletes who were "born, live, and train at altitude" or just elite athletes in general are so different physiologically or biologically that it makes sense to assume that their bodies respond differently. In the absence of clinical studies, it makes sense to believe their bodies respond the same way everybody else's do.

+1

jingle jingle wrote:

Merry Christmas Mr Canova!

rjm33,You told us there was ZERO evidence. Looks like you were wrong.

rjm33 wrote:

...You have ZERO evidence to back up your statement on increased affinity of hemoglobin for oxygen in altitude responders.

rekrunner wrote:

No one ever said how much 2:08 Kenyans would improve, if at all.

Correct. However, Renato explained that EPO works normally (like for Ramzi, and for Aden's group except for Dibaba), but not for the subclass of top Kenyans born, living and training at altitude. Then he proceeded to call 2:08 Kenyans mid-level Kenyans, so I was trying to verify how much, in Renato's opinion, a "medium athlete" could improve with EPO.

Renato Canova wrote:

The fact that ALL THE BEST RECORD HOLDERS were able to run completely clean (some of them is/was my athlete, so I well know)

Those are two very bold statements. I am 100% that the first one is wrong however. For examples see Junxia (3000 m WR) and Liliya (30 km road WR). (You are welcome, rekrunner.) But if Renato meant to limit that to these altitude born/raised/trained athletes, then there are no counterexamples (yet).

The second one, i.e. that coaches know whether or not their athletes dope (which is obviously correct for capable coaches), shines a certain light onto the coaches of drug cheats. And on athletes who chose to train under coaches of drug cheats.

Not doped and shafted at the Olympics

http://therecord.blogs.com/.a/6a00d8341c465d53ef016764e6c4c8970b-600wi

I don't think any Kenyan can have some real advantage taking EPO, if decides to change his training increasing the volume of specific quality.

I know personally athletes running for 3-4 years marathon in 2:12 - 2:14, with little and wrong training, and without the possibility to eat more than once per day. After this situation, they were invited in some training camp managed by some strong athlete (for example, Dennis Kimetto in the camp of Geoffrey Mutai), started to eat 3 times per day, started to have different training with more volume and more intensity, and immediately ran 2:07.

Athletes who are not in some advanced training group, especially who continues to live in his village, don't train really hard, and particularly don't have any progression in their training, always repeating the same schedule. In other words, they "run" but don't "train", since running and training are two different things. They are "amateurs" as type of training, interest in running and mental motivation, with the difference that an Italian amateur of good level runs 2:20, the Kenyan 2:13.

In some case, if we explain to athletes alone in their village how much training they have to do, and with which intensity, in order to become able running 2:07, they see themselves, at the moment, so far from that situation, that start thinking of some external help, in order to change their training.

Taking EPO gives them the mental strength for starting to think possible a training looking at that performance, while without this illegal support they think that NEVER can train so hard (and this was the motivation of Mathew Kisorio for asking for EPO and steroids).

So, finally certain in their mind that now can be able to train so hard, recovering the fatigue, they start the REAL TRAINING OF HIGH LEVEL for the first time in their life.

It's the training pushing them to run 2:07, not EPO, but EPO is the reason because, for the first time, they accept to train so hard.

It's possible to train so hard also without EPO, if the mind is from the beginning n this direction, and this is the first basic difference between top champions and good athletes : top champions don't fear training and competition, and want to explore everytime new horizonts, while good athletes fear to go in a new (for them) situation, preferring to remain inside the limits of training already known.

I'm not sure that high Hct and high Hb can be a reason for improving the PB of an athlete. There is something different that can explain the exploits of the best. If I coach an athlete who continues to improve and I carry out with him periodical blood tests, I can see that, when his shape is better and he is able running faster, his values go down, and Hct and Hb are lower than when he is in little training and not able running fast. This is a NORMAL condition we can always find in ALL the athletes, including who always train at sea level, and frankly I don't understand how this situation can be explained by the idea that higher Hct and Hb, better performance.

I think that with EPO, continuining the same type of training, with the same intensity and the same volume of BEFORE EPO, we can have a little increase in performances. But the reality is that all the athletes taking EPO increased their training in all the directions, giving merit to the EPO assumption for the possibility to increase their training, BUT NEVER THEY TRIED TO HAVE THE SAME KIND OF TRAINING WITHOUT EPO, BECAUSE SUPPOSED IT WAS NOT POSSIBLE.

At the end, their improvement is directly due to the increase of their training in both direction (volume and intensity). The real question, therefore, is : IS EPO DIRECT, OR NOT DIRECT, RESPONSIBLE FOR THE INCREASE OF QUALITY IN TRAINING ?

Many researchers give merit to EPO looking at the variations of the blood values. I, instead, think that the blood values are EFFECT, not CAUSE, of training, and the factor changing the level of performances is a different training only, so the real effect of EPO was only to give more confidence to the athlete, now accepting the idea to train in more hard and smart way.

This argument keeps getting better and better.

Athletes take EPO, train harder, and race faster.

But it is not because of the EPO that they run faster.

Catch-22