635 replies to this topic

[Turnbuckle]

That's what I proposed in post 313 of this thread, except that the example I used was a related compound, SkQ1. I still think this is a plausible explanation, mainly hinging on the ability of C60 to react with unsaturated fatty acids under mild conditions, which it appears to do. SkQ1 is reported to be good at squaring the lifespan curve in rodents, which C60/olive oil is extremely good at, if we believe what we read in the paper...

Except that MitoQ is supposed to be a CoQ10 analogue, which itself is supposed to be a super antioxidant that is delivered preferentially to the mitochondria, yet has been shown not to increase the lifespan of rodents.

[Turnbuckle]

I wonder. According to the theories (on how C60 might work) mentioned in this forum, what would an old person (say, over 80) expect by taking C60 with olive oil? I mean, whould this person expect improvement of just a delay of what is next to come? In other words, is it logical to assume that by taking C60 with olive oil (assuming of course that the rat study is correct and that it can be applied in humans exactly as it is), the old person will experience a "becoming younger" effect?

If the effect is in the mitochondria, as it appears to be, then that will gradually be reflected in the systems of the body. The first results will likely appear in those organs that recycle the fastest--the gut, skin, and liver. At the age of sixty, I've noticed lower cholesterol and much better alcohol tolerance, and also better skin and some hair regrowth. So there's definitely some global reshaping to a more youthful age. How far that will go, I don't know, but I expect it's limited. I won't become Benjamin Button, but I'll settle for being carded once in a while.

[niner]

The explanation for that Olive Oil only study result has already been figured out. I posted about it in post #501.

Well, that would explain why people on high fat diets feel better than when on low fat diets. In Luyer's paper, which Tracy references, they used 52 energy% fat for the high fat diet, and 17% for the low fat diet in rodents. Two ml oil per kg body weight would be enough to trigger the CAIP, at least in rats if not humans, but wouldn't that be a transitory effect? I don't see how doing that 24 times in an animals lifetime would cause such an extreme life extension.

[niner]

Except that MitoQ is supposed to be a CoQ10 analogue, which itself is supposed to be a super antioxidant that is delivered preferentially to the mitochondria, yet has been shown not to increase the lifespan of rodents.

But C60 is a super antioxidant too. Super-er than Q10, even. It's a SOD mimetic. There's a big difference between ordinary Q10 and MitoQ, because MitoQ is targeted directly to the mitochondrial membrane and held there, while ordinary Q10 floats around wherever it wants.

Sure seems there's something to high dose olive oil for inflammation. Dang, my Italian Dad has been touting the stuff for years!

Jeanne Calment, too.

[Turnbuckle]

But C60 is a super antioxidant too. Super-er than Q10, even. It's a SOD mimetic. There's a big difference between ordinary Q10 and MitoQ, because MitoQ is targeted directly to the mitochondrial membrane and held there, while ordinary Q10 floats around wherever it wants.

Still, quite a lot of CoQ10 goes to the mitochondria, so you would still expect that supplementation would result in a longer lifespan if its antioxidant character were a factor. But it didn't.

On the other hand, using olive oil as a vector to take more of it into the mitochondria ought to help whatever it does do: Olive oil supplemented with Coenzyme Q(10) : Effect on plasma and lipoprotein oxidative status.

"EV olive oil enriched with both doses of CoQ(10) significantly affects its bioavailability and plasma redox status. These changes are associated with a decreased susceptibility of plasma lipoproteins to peroxidation associated with a chain-breaking antioxidant activity of the formulation."

Edited by Turnbuckle, 28 May 2012 - 12:57 PM.

[niner]

Still, quite a lot of CoQ10 goes to the mitochondria, so you would still expect that supplementation would result in a longer lifespan if its antioxidant character were a factor. But it didn't.

Not really. CoQ10 has lousy bioavailability, for one thing, and MitoQ, because it contains a hydrophobic cation, drags the Q10 moiety into the mitochondrial membrane, driven by the large membrane potential. The result is hundreds of times more antioxidant in the mitochondria, so they really aren't comparable at all.

On the other hand, using olive oil as a vector to take more of it into the mitochondria ought to help whatever it does do: Olive oil supplemented with Coenzyme Q(10) : Effect on plasma and lipoprotein oxidative status.

The olive oil is definitely enhancing the bioavailability of the Q10. That's a general phenomenon of hydrophobes in oil versus trying to get them absorbed from a dry state. However, the oil doesn't act like a chaperone, bringing Q10 into the mitochondrion. By the time the mixture has been absorbed to that extent, the oil and the Q10 will be fairly well separated.

In order to act as a chaperone, the two molecules would either need to be covalently linked, or one of them would need to be an "artificial receptor" for the other, like the way certain macrocycles, like crown ethers or cyclic peptides, can hold a cation, or the way a cyclodextrin can hold and solublize a hydrophobe. When Q10 is dissolved in olive oil, the big difference compared to dry Q10 is that it's dissolved instead of crystalline, and that it follows the oil into lymphatic circulation. Ultimately, though, the Q10 goes its own way, riding on various lipid carrier proteins and accumulating in various lipid depots throughout the organism, some fraction of which will be mitochondria.

[JohnD60]

Playing devil's advocate for a moment, these guys http://www.highbeam....-500897071.html have been looking at this tech for close to 10 years. So why no fullerene drugs or supplements already? Do they work too well, not at all, or too much risk for healthy patients (as in buy the innovator C-Sixty and then bury the work).

I am not sure from that blurb how much actual research was done on the subject. My belief is that Big Pharama is profit driven and their research is steered toward profitiable projects. I do not see a lot of profit potential in researching C60 forumulations because they can not patent C60. At best they could patent some formulation that includes C60, but that formulation could be easily hacked. So they end up with a product that has all the liabilty of their usual products, but a fraction of the profit potential. And, the cynical side of me thinks that broad range cures or life extension is not in the best interest of Big Pharama as it would cannbalize their existing profits centers.

[niner]

as another poster said I am leaning toward the theory that the c60 does " wipes out the methylation, the DNA automatically recovers it, and the effective age is set to zero."

I'm pretty sure that even the poster who proposed this doesn't buy it anymore. The odds of this being the mechanism of action are vanishingly small, as it requires several highly improbable steps, one of which might kill you.

[Turnbuckle]

I'm pretty sure that even the poster who proposed this doesn't buy it anymore. The odds of this being the mechanism of action are vanishingly small, as it requires several highly improbable steps, one of which might kill you.

Actually, I do. Even though mitochondria tend to lose global methylation with age--which seemingly goes against the theory--they also tend to get the methylation pattern scrambled, and that may be even more deleterious.

[HighDesertWizard]

I'm pretty sure that even the poster who proposed this doesn't buy it anymore. The odds of this being the mechanism of action are vanishingly small, as it requires several highly improbable steps, one of which might kill you.

Actually, I do. Even though mitochondria tend to lose global methylation with age--which seemingly goes against the theory--they also tend to get the methylation pattern scrambled, and that may be even more deleterious.

There are other extremely healthful substances, the ones I know about 5-Lipoxygenase Inhibitors, which have been shown to do demethylation of DNA, Boswellia and Curcumin for example.

Boswellic acid induces epigenetic alterations by modulating DNA methylation in colorectal cancer cells

I once experienced a dramatic change in my worsening Carpal Tunnel Syndrome symptoms within a week of ingesting very high dose Boswellia (AKBA). I stopped the ~5 days of dosing and the symptoms returned. I then restarted the very high dosing and the symptoms vanished again and after a couple of weeks I stopped again. And the symptoms have never returned.

I have always wondered "what the heck happened?"

Turnbuckle's experience has been interesting to me because of how rapidly his positive health change took place. Even more rapid than mine did.

The open question is: What is the explanation for this kind of positive change this quickly?

Edited by wccaguy, 30 May 2012 - 06:01 PM.

[HighDesertWizard]

Here is one of the original Turnbuckle statements providing an explanation for the positive and long lasting health effects of C60 in the study.

Assuming my hypothesis that C60 acts to strip methyl groups from the mitochondrial DNA is correct, then curcumin (which I've taken for some time) might act synergisticly--

Quote

Genomic DNA was extracted from a leukemia cell line exposed to curcuminoids at concentrations of 0, 1, 3, and 30 μM for 72 hours. Analyses of global DNA methylation levels showed stability at 1 μM curcumin, but decreased by approximately 15% to 20% at 3 μM and 30 μM curcumin compared with untreated basal methylation levels, which was equivalent to decitabine-induced decreases in global DNA methylation levels. These data show that curcumin is a potent DNA hypomethylating agent, which is consistent with its broad activity in inflammation, cancer, and many other diseases, while remaining relatively safe in normal healthy cells.

Development of curcumin as an epigenetic agent.

There are other extremely healthful substances, the ones I know about, 5-Lipoxygenase Inhibitors, which have been shown to do demethylation of DNA, Boswellia and Curcumin for example.

Boswellic acid induces epigenetic alterations by modulating DNA methylation in colorectal cancer cells

5-Lipoxygenase Inflammatory Pathway inhibitors both inhibit inflammation and "modulate" gene level processes. I posted just one study link to avoid cluttering the thread with dozens.

I'm pretty sure that even the poster who proposed this doesn't buy it anymore. The odds of this being the mechanism of action are vanishingly small, as it requires several highly improbable steps, one of which might kill you.

Actually, I do. Even though mitochondria tend to lose global methylation with age--which seemingly goes against the theory--they also tend to get the methylation pattern scrambled, and that may be even more deleterious.

In Turnbuckle's case, he had what sounds like a pre-existing mitochondrial dysfunction. It's not surprising to me that this could be rapidly corrected with a redox-active compound (just like his experience with PQQ). With your carpal tunnel syndrome, you might have broken an inflammatory feedback loop; the same thing might have happened with a different anti-inflammatory compound. Typically, if you can get inflammation to calm down, it goes away on its own.

The first hypothesis should be the simplest and most plausible.

My questions are these...

The same highly improbable steps required for C60 to act at the level of the gene would be required of 5-LO inhibitors. How do we know for certain, then, that this isn't the mechanism of action of C60? Why isn't the simplest explanation of C60 action the same as that increasingly found for 5-LO inhibitors?

I may be confused but I think we've been circling around the big question here, namely, the apparent long lasting effect of C60 action given that the authors state it cleared within a few "tens of hours." It's legitimate to argue that they must have screwed up the clearance data. They did screw up some graphic figures evidently. But the simplest explanation is not to assume they screwed the clearance data too.

Again, I apologize if my limited understanding of these issues is creating needless churn... It seems to me, however, that, so far, Turnbuckle's hypothesis explains a lot. It...

• explains the long lasting effect of the C60s in the rats
• explains the virtually immediate positive impact on his own health
• is consistent with increasing evidence about 5-LO inhibitor impact taking place at the gene level

Maybe it's been posted before and I missed it...

What is the alternative explanation for both the long lasting and immediate positive effect of C60, other than to suggest that the study authors screwed up the C60 clearance data?

One final question...

As one component of any followup projects Longecity might do, what experiments might be performed to confirm the C60 gene level impact hypothesis? More importantly, how might we Falsify it?

Edited by wccaguy, 31 May 2012 - 04:56 AM.

[niner]

Again, I apologize if my limited understanding of these issues is creating needless churn... It seems to me, however, that, so far, Turnbuckle's hypothesis explains a lot. It...

• explains the long lasting effect of the C60s in the rats
• explains the virtually immediate positive impact on his own health
• is consistent with increasing evidence about 5-LO inhibitor impact taking place at the gene level

Maybe it's been posted before and I missed it...

What is the alternative explanation for both the long lasting and immediate positive effect of C60, other than to suggest that the study authors screwed up the C60 clearance data?

Turnbuckle's hypothesis was that C60 stripped all the methyl groups off the DNA, and that these were 'rebuilt' to a youthful state. That can't be right; nearly all the methyl groups are inaccessible most of the time, there is no known chemistry whereby C60 would abstract a methyl group, and there would be a period of time where gene expression was completely screwed up, probably lethally, and I don't think that somatic cells can do this rebuilding thing anyway, further, there is no known epigenetic change that results in a 90% increase in lifespan. As David Deutsch said in a Ted talk you posted recently, a good hypothesis must be hard to vary. This one can be varied all over the map: e.g.: C60 hypermethylates the DNA, which is then de-methylated to a youthful state. It's not methyl groups, it's acetyl groups. It's not either of them, it's modifying the histones. It's not modifying the histones, it's replacing them, resulting in a more efficient genome...

So maybe C60 is interacting with one of the various enzymes that control epigenetic modifications? This is at least plausible, but C60 is a large hydrophobic sphere. To interact with an enzyme, it would need a large hydrophobic spherical cavity, and that is not the sort of thing that you find very often in proteins. And this still doesn't explain how an epigenetic change could have such a large effect on lifespan. If you ask Aubrey de Grey, he would tell you that somatic cell mutation and epimutation are not significant contributors to aging, and only matter in the case of cancer.

Instead, let's start with things that we know to be true about C60: It acts as a "free radical sponge", soaking up electrons from radicals, holding them in a stable form, and giving them up to reactions that produce safer products. One consequence of this is that it can act as an SOD mimetic, performing a dismutation of the dangerous superoxide ion which is produced during mitochondrial respiration. We have spectroscopic evidence that C60 forms adducts with vegetable oils. Cataldo says that they are formed at room temperature over a period of days when the oil/C60 mixture is exposed to air. They are red in color, like the solutions that people are seeing. They are structurally and electronically similar to mitochondrially active agents like MitoQ or SkQ1. I propose that they work the same way as those agents; they become part of the mitochondrial membrane where they neutralize damaging free radicals.

That's my explanation for the immediate effects; here is an explanation that explains the long term effects and addresses the clearance issue:

The clearance data is only showing what is excreted from the rat, but not what stays inside. It's not a quantitative recovery of C60. I propose that the 24 treatments loaded the rats' membranes with C60 fatty acid adducts. As mitochondria are turned over through autophagy, the C60 compounds are released and are recycled into a new membrane. This could be tested by using a radiolabeled C60, so you could determine the time course of its residence in the animal.

[HighDesertWizard]

Turnbuckle's hypothesis was that C60 stripped all the methyl groups off the DNA, and that these were 'rebuilt' to a youthful state. That can't be right; nearly all the methyl groups are inaccessible most of the time, there is no known chemistry whereby C60 would abstract a methyl group, and there would be a period of time where gene expression was completely screwed up, probably lethally, and I don't think that somatic cells can do this rebuilding thing anyway, further, there is no known epigenetic change that results in a 90% increase in lifespan. As David Deutsch said in a Ted talk you posted recently, a good hypothesis must be hard to vary. This one can be varied all over the map: e.g.: C60 hypermethylates the DNA, which is then de-methylated to a youthful state. It's not methyl groups, it's acetyl groups. It's not either of them, it's modifying the histones. It's not modifying the histones, it's replacing them, resulting in a more efficient genome...

So maybe C60 is interacting with one of the various enzymes that control epigenetic modifications? This is at least plausible, but C60 is a large hydrophobic sphere. To interact with an enzyme, it would need a large hydrophobic spherical cavity, and that is not the sort of thing that you find very often in proteins. And this still doesn't explain how an epigenetic change could have such a large effect on lifespan. If you ask Aubrey de Grey, he would tell you that somatic cell mutation and epimutation are not significant contributors to aging, and only matter in the case of cancer.

Instead, let's start with things that we know to be true about C60: It acts as a "free radical sponge", soaking up electrons from radicals, holding them in a stable form, and giving them up to reactions that produce safer products. One consequence of this is that it can act as an SOD mimetic, performing a dismutation of the dangerous superoxide ion which is produced during mitochondrial respiration. We have spectroscopic evidence that C60 forms adducts with vegetable oils. Cataldo says that they are formed at room temperature over a period of days when the oil/C60 mixture is exposed to air. They are red in color, like the solutions that people are seeing. They are structurally and electronically similar to mitochondrially active agents like MitoQ or SkQ1. I propose that they work the same way as those agents; they become part of the mitochondrial membrane where they neutralize damaging free radicals.

That's my explanation for the immediate effects; here is an explanation that explains the long term effects and addresses the clearance issue:

The clearance data is only showing what is excreted from the rat, but not what stays inside. It's not a quantitative recovery of C60. I propose that the 24 treatments loaded the rats' membranes with C60 fatty acid adducts. As mitochondria are turned over through autophagy, the C60 compounds are released and are recycled into a new membrane. This could be tested by using a radiolabeled C60, so you could determine the time course of its residence in the animal.

niner... I don't yet understand your reply in its entirety. I have homework to do to understand it all. Nevertheless, I think it's one of the most informative and insightful posts in this entire thread and moves the discussion forward significantly. Thank you.

[Turnbuckle]

The clearance data is only showing what is excreted from the rat, but not what stays inside. It's not a quantitative recovery of C60. I propose that the 24 treatments loaded the rats' membranes with C60 fatty acid adducts. As mitochondria are turned over through autophagy, the C60 compounds are released and are recycled into a new membrane. This could be tested by using a radiolabeled C60, so you could determine the time course of its residence in the animal.

We discussed that before and I pointed out that even a small loss rate would eliminate all of the C60 in short order. For instance, the average life of a rat liver mitochondria is between one and two days, so after six months it would undergo something like 100 mitochondrial fissions--ie, 100 generations. The half lives of mitochondrial enzymes and proteins are known from C14 studies to be between 1 day and 1 week--similar to what was reported in the rat paper for C60 where all of it was eliminated in "a few tens of hours"--so if the C60 half life were actually as long as a week, the mitochondria would lose 99.999999% of it after six months, and most mitochondria would not have even a single molecule of C60. And even if the mitochondria lost only 5% of the original C60 in each generation (an unbelievably small amount), the final generation after six months would still have lost 99.5% of what it had originally.

Now given that the rats were dosed over a period of six months and given that almost none of the C60 would be around for very much longer, the best the rats could have hoped for would be a life extension of less than twelve months, and more like six or seven months. Yet they got a lot more than that, so the actual mechanism has to be something other than (or in addition to) C60 acting as a resident super anti-oxidant.

Edited by Turnbuckle, 31 May 2012 - 05:55 PM.

[hav]

If the C60 isn't excreted, I would presume its still in there somewhere doing its anti-oxidizing thing. Unless there's some other accounting for where it went.

Howard

Edited by hav, 31 May 2012 - 06:40 PM.

[Turnbuckle]

If the C60 isn't excreted, I think your analysis would need to explain where it went. Otherwise, I would presume its still in there somewhere doing its anti-oxidizing thing.

From the rat paper--

The elimination process follows a non-urinary route because
unmodified C60 was not detected in urine samples taken up 48 h
after administration. Previous investigations showed that C60 is
mainly eliminated through the bile ducts as it has been
recently confirmed. Besides, a small increase in C60 concentrations
at 12 and 24 h after i.p. administration (Fig. 1) suggests the
presence of an enterohepatic circulation. Furthermore, it has
been already shown that C60 reacts inside the liver cells with
vitamin A following a DielseAlder like reaction both in mice and in
rats. These two routes may be sufficient for C60 elimination,
nevertheless, we have to look for other possible biotransformations
and elimination routes, all the more so as the fate of the addition
product is not known...Nevertheless,
the weakness of organ concentrations notably at D8 after 7
daily successive administrations of C60 dissolved in olive oil clearly
shows that C60 molecules are eliminated from the organs in a few
hours after both oral and i.p. administrations.

Edited by Turnbuckle, 31 May 2012 - 06:46 PM.

[hav]

Sounds like all the elimination routes they postulate lead to the stool. Do you think any could be exhaled?

Howard

[Turnbuckle]

Sounds like all the elimination routes they postulate lead to the stool. Do you think any could be exhaled?

Howard

Since C60 sublimes at nearly 1000 F, I don't think so.

[taho]

What happens to c60 in those C60 fatty acid adducts if they are used as fuel in mitochondria?

My thinking is.. if they would "fall out" and "hang around" there, they could reduce mitochondria autophagy by soaking up free radicals that signal which mitochondria are broken and need to be replaced. That interferance could reduce long term acumulation of damaged mitochondria. Kind of long shot, but it could explain why people with damaged mitochondria would get instant efect. The body stops destroying them faster then their replacements could be grown.

The second line of thought is, that some of it ends up incorporated into the cell lipid walls, that has much longer half life and basicly catches ROS before they can destroy (poly?) unsaturaded lipids, and that stops ROS chain reaction and all the damage that results from it. Since c60 is not destroyed by ROS, it will work as long as it is in the body (anywhere in the body). And since it crosses the blood-brain barrier, that means less destruction of nevrons, which means that the body can build new things not fight a progresivly loosing battle to repair ROS damage..

[niner]

We discussed that before and I pointed out that even a small loss rate would eliminate all of the C60 in short order. For instance, the average life of a rat liver mitochondria is between one and two days, so after six months it would undergo something like 100 mitochondrial fissions--ie, 100 generations. The half lives of mitochondrial enzymes and proteins are known from C14 studies to be between 1 day and 1 week--similar to what was reported in the rat paper for C60 where all of it was eliminated in "a few tens of hours"--so if the C60 half life were actually as long as a week, the mitochondria would lose 99.999999% of it after six months, and most mitochondria would not have even a single molecule of C60. And even if the mitochondria lost only 5% of the original C60 in each generation (an unbelievably small amount), the final generation after six months would still have lost 99.5% of what it had originally.

Yes, if they lose that much, but I don't think they do. The mitochondria are inside the cell, and mitophagy occurs entirely within the cytoplasm. The C60-fatty acid adduct wouldn't have a rapid way to leave the cell I doubt very much that the lysosome is able to degrade C60, so the C60 probably remains intact even if the adduct doesn't, though I suspect the adduct would remain intact as well. I wouldn't expect very much at all to be lost. If 1% were lost per generation, after a hundred turnovers, there would be 37% of the original amount remaining. If 0.1% were lost, there would be 90% left. We don't actually know how much C60 you would need in the membrane to get the job done, so the initial amount might be in substantial excess. I would speculate that humans would need to be "topped up" every so often; maybe once a year, more or less?

[maxwatt]

...
Instead, let's start with things that we know to be true about C60: It acts as a "free radical sponge", soaking up electrons from radicals, holding them in a stable form, and giving them up to reactions that produce safer products. One consequence of this is that it can act as an SOD mimetic, performing a dismutation of the dangerous superoxide ion which is produced during mitochondrial respiration. We have spectroscopic evidence that C60 forms adducts with vegetable oils. Cataldo says that they are formed at room temperature over a period of days when the oil/C60 mixture is exposed to air. They are red in color, like the solutions that people are seeing. They are structurally and electronically similar to mitochondrially active agents like MitoQ or SkQ1. I propose that they work the same way as those agents; they become part of the mitochondrial membrane where they neutralize damaging free radicals.

That's my explanation for the immediate effects; here is an explanation that explains the long term effects and addresses the clearance issue:

The clearance data is only showing what is excreted from the rat, but not what stays inside. It's not a quantitative recovery of C60. I propose that the 24 treatments loaded the rats' membranes with C60 fatty acid adducts. As mitochondria are turned over through autophagy, the C60 compounds are released and are recycled into a new membrane. This could be tested by using a radiolabeled C60, so you could determine the time course of its residence in the animal.

I would like to raise the possibility that it is the polyphenols in extra virgin olive oil that are responsible for the observed effect, and that C60 is acting as a carrier or a chaperone for those polyphenols, increasing their efficacy. If I recall, the olive-oil-only control group rodents also showed unusual longevity, though it was overshadowed by that of the C60+olive oil group. This is consistent with the hypothesis that C60 is a carrier for EVOO polyphenols.

[niner]

I would like to raise the possibility that it is the polyphenols in extra virgin olive oil that are responsible for the observed effect, and that C60 is acting as a carrier or a chaperone for those polyphenols, increasing their efficacy. If I recall, the olive-oil-only control group rodents also showed unusual longevity, though it was overshadowed by that of the C60+olive oil group. This is consistent with the hypothesis that C60 is a carrier for EVOO polyphenols.

Yes, actually the olive oil-only group had a rather shockingly large increase in lifespan, which is kind of hard to explain. Can it really be that no one ever used an intermittent dosing strategy, and that's the magic? Or is this just indicative of a whole raft of errors, of which the olive oil-induced LS increase is just the tip of the iceberg? Or is this the first time someone used a "really good" olive oil? I have to admit, the error idea is pretty attractive here... At any rate, a factor of almost two in LS from EVOO polyphenols is a lot to swallow; surely the fullerenes were also doing something, wouldn't you think? Finally, C60 is a pretty tight cage; I don't see how a typical polyphenol would get inside. I can't imagine any other chaperone mode that makes much structural sense.

[maxwatt]

It is a conundrum, an enigma wrapped in a buckyball.

Perhaps only a piece of a polyphenol gets attached to a fullerene molecule, and transported thusly? Hard to swallow, but there are patents out there for using fullerenes to transport substances in this manner. C60 also forms macro-structures of several bucky-balls, in a crystal-like arrangement even in acqueous solutions, and these can trap other, somewhat larger molecules. The possibilities are many, each seemingly equally implausible.

I would not have suggested it but for Michael's enthusiasm for EVOO polyphenols. and their possible life-enhancing effects. They are poorly understood, we don't know what molecules are present, much less which of them might be responsible for the phenomenon we are investigating. But EVOO polypheneols do inhibit HER-2, killing cancer cells, they inhibit inflammation and tyrosine kinase also. That the oil-only rodents also exhibited an unusual increase in lifespan lends weight to the hypothesis. I don't insist the hypothesis is true, but think it quite possible than we are seeing an interaction between the olive oil and the fullerene.

In the meantime, I would suggest using the best quality, polyphenol-rich olive oil you can find if you want to play with fullerenes.

Or it is possible and even likely that the results are not reproducible.

[HighDesertWizard]

About C60 clearance...

... if the mitochondria lost only 5% of the original C60 in each generation (an unbelievably small amount), the final generation after six months would still have lost 99.5% of what it had originally.

From the rat paper--

...Nevertheless, the weakness of organ concentrations notably at D8 after 7

daily successive administrations of C60 dissolved in olive oil clearly
shows that C60 molecules are eliminated from the organs in a few
hours after both oral and i.p. administrations.

Yes, if they lose that much, but I don't think they do...

About the profound difference Olive Oil appears to have made...

Yes, actually the olive oil-only group had a rather shockingly large increase in lifespan, which is kind of hard to explain. Can it really be that no one ever used an intermittent dosing strategy, and that's the magic? Or is this just indicative of a whole raft of errors, of which the olive oil-induced LS increase is just the tip of the iceberg?

I don't think any of us have the time and energy to go through a dozen papers on the Vagus--CAIP--HRV nexus on the thin chance that it's relevant to the reported effects of C60. Remember, there is a large difference in lifespan enhancement between the olive oil control and the C60/olive oil mixture. I don't see how the vagal mechanism explains that.

Heart Rate Variability (HRV) has been a subject of at least 1000 scientific studies over more than 20 years. It's not some new scientific study subject. I posted the following study as well as others making the same point about the importance of HRV to Longevity here at post 551. I posted links to 25 studies showing the importance of HRV to Longevity and Disease incidence here.

---------------------------
Relation of high heart rate variability to healthy longevity

The population's aging underscores the need to understand the process and define the physiologic markers predictive of healthy longevity. The findings that aging is associated with a progressive decrease in heart rate variability (HRV), an index of autonomic function, suggests that longevity might depend on preservation of autonomic function. However, little is known about late life changes.... The HRV-sympathetic function continues to decrease throughout life. In contrast, the decrease in HRV-parasympathetic function reaches its nadir in the eighth decade, followed by reversal and a progressive increase to higher levels (p <0.05), more characteristic of a younger population. In conclusion, healthy longevity depends on preservation of autonomic function, in particular, HRV-parasympathetic function, despite the early age-related decrease. The eighth decade reversal of the decrease in HRV-parasympathetic function and its subsequent increase are key determinants of longevity. Persistently high HRV in the elderly represents a marker predictive of longevity.
---------------------------
Here's a graphic figure from that study. The X axis is defined by study population age bracket. The Y axis defines HRV-related data points about the study populations who have survived into the X axis age bracket.



---------------------------

I posted links to multiple studies in another thread that convincingly demonstrates the existence of an Innate Immune System related process called the Cholinergic Anti-Inflammatory Pathway (CAIP).

I posted links to studies here in post 501 that shows 1) Vagus Nerve Stimulation Activates the CAIP, 2) Dietary Fat Stimulates the Vagus Nerve, and 3) Olive Oil is an especially powerful Stimulator of the Vagus Nerve.

But the Vagus Nerve can be stimulated by more than just Olive Oil. And the CAIP includes specific, well understood, measurable, life extension-related impacts on HRV.

Vagus Activation -->> Higher HRV: http://www.jci.org/articles/view/30555

Positive Emotions -->> Extreme Longevity: http://www.impactagi...bs/100456a.html
Positive Emotions -->> Vagus Activation: http://www.ncbi.nlm....pubmed/20851735

Exercise -->> Extreme Longevity: http://extremelongev...ds/sardinia.pdf
Exercise -->> Vagus Activation: http://www.ncbi.nlm....pubmed/11560079
Respiratory muscle training -->> Improves "sympathetic and vagal heart modulation": http://jap.physiolog...11/6/1664.short

----------------------------------------

A final point... the CAIP inhibits TNF-a and IL-6 expression when "Vagal Tone" is high, among other things... So the CAIP impact ROS, mitochondria, etc. For example...

http://www.ncbi.nlm....pubmed/20203691

----------------------------------------

Late last year, I was astonished when I first learned of the CAIP. What caused my astonishment? It was my ego. I'm 57 and I had been a very serious student of Inflammatory Diseases and processes for several years. Kevin Tracey had first written about the CAIP in 2002 and it was 2011. And I couldn't believe something as important as the CAIP had escaped my notice all that time...

As the acceleration of the growth of scientific knowledge continues, we can EXPECT new ideas and new evidence will race toward us. So we all face a constant struggle to keep our game faces on and to remain open to new evidence and ideas, even if they don't fit our preferred categories, theories, and preferred ways of thinking.

I expect to get stuck in a rut sometimes as the new evidence races toward me. And I've found that the probability of my having gotten stuck is high when I must resort to insisting that the data is wrong rather than explore the notion that my preferred theory can't explain everything...

Edited by wccaguy, 01 June 2012 - 11:45 AM.

[HighDesertWizard]

It [the question of Olive Oil's contribution to health] is a conundrum, an enigma wrapped in a buckyball.

Um, no, it's not. It's the Olive Oil -->> CCK -->> Vagus Nerve -- >> CAIP/HRV Nexus and Kevin Tracey is going to win the Nobel Prize in Medicine for doing the most work around it...

8-)

Edited by wccaguy, 01 June 2012 - 11:14 AM.

[niner]

HRV is associated only very loosely with normal aging. Look at the boxplots you posted; the means and medians barely change at all between the 8th and 9th decade, only the outliers. But you see the next highest outliers in the third decade! I agree that the CAIP is important in normal humans, but I've seen no evidence at all that it is involved in abnormally long lifespan.

[HighDesertWizard]

HRV is associated only very loosely with normal aging. Look at the boxplots you posted; the means and medians barely change at all between the 8th and 9th decade, only the outliers. But you see the next highest outliers in the third decade! I agree that the CAIP is important in normal humans, but I've seen no evidence at all that it is involved in abnormally long lifespan.

Have you browsed the other HRV related abstracts I provided links to? If you had, you'd have come across this.

Altered autonomic regulation of cardiac function may contribute to the onset of cardiovascular disease and provide a substrate for malignant ventricular arrhythmias. This study was designed to assess cardiovascular neuroautonomic status in healthy subjects with short-term power spectral analysis of heart rate variability, including a group over 100 years of age, to identify a neuroautonomic pattern that could help to protect ultra-centenarians against cardiovascular disease. One hundred and twelve subjects (22 men and 90 women, age range 20 to 107 years) were subdivided into five age groups: <40 years (N=26, mean age 30.6±0.9); 41 to 60 years (N=27, mean age 51.9±1.2); 61 to 80 years (N=37, mean age 70.3±1.1); 81 to 100 (N=10, mean age 85.2±0.8) and older than 101 years (N=13, mean age: 103.6±0.6). Power spectral analysis with autoregressive algorithm provides two indexes of autonomic activity: a low-frequency component oscillating around 0.10 Hz, mainly reflecting sympathetic activity and a high-frequency component around 0.30 Hz, reflecting parasympathetic activity. Subjects 40 years of age or younger had significantly higher spectral high-frequency power values expressed in logarithmic form than the other age groups (P<0.05), the age group from 41 to 100 years had values similar to those of the other groups. However, the age group over 101 years had significantly higher values than the group from 81 to 100 years (P<0.05). Low-frequency spectral density expressed in logarithmic form and in normalized units decreased with age (P<0.0001). These data confirm an age-related decline in sympathetic activity. Compared with elderly subjects from 81 to 100 years of age ultra-centenarians have significantly higher spectral parasympathetic indexes. Parasympathetic predominance may be the neuroautonomic feature that helps to protect ultra-centenarians against cardiovascular disease.

→ source (external link)

Imagine what those boxplots in that figure would have looked like given this data. If the results of those two studies were combined, the boxplot for 100 and higher would have been higher than for 80-100...

An implication of the study above is that those outliers in the 80-100 data on the high side are the survivors when the 100+ bracket is hit.

Can we agree now that you have seen some, maybe just a smidgen of, evidence that CAIP is involved in abnormally long lifespans?

HRV is the only age related statistic I know of that does an abrupt about face at about the age 80 among survivors within various age brackets... Do you know of another age related statistic that does an about face like that?

Finally, after review of this evidence, do you believe it makes more sense and is a simpler explanation that Baati et al have gotten the data of their study profoundly wrong with regard to Olive Oil than to take the CAIP as being important? Especially given the fact that Olive Oil has been shown to more powerfully trigger the CAIP than most (all?) other dietary substances in the gut?

(I assume here that Anthony Loera heard it right that Baati et al understood that they had some graphic figures wrong but the data in the text was right.)

Edited by wccaguy, 01 June 2012 - 12:34 PM.

[Turnbuckle]

HRV is associated only very loosely with normal aging. Look at the boxplots you posted; the means and medians barely change at all between the 8th and 9th decade, only the outliers. But you see the next highest outliers in the third decade! I agree that the CAIP is important in normal humans, but I've seen no evidence at all that it is involved in abnormally long lifespan.

I agree. This Vagus stuff is very left field. In the meantime, we can rejoice that the two oldest people in the world until recently (both over 120) credited their longevity in part to olive oil.

Edited by Turnbuckle, 01 June 2012 - 12:30 PM.

[niner]

Can we agree now that you have seen some, maybe just a smidgen of, evidence that CAIP is involved in abnormally long lifespans?

HRV is the only age related statistic I know of that does an abrupt about face at about the age 80 among survivors within various age brackets... Do you know of another age related statistic that does an about face like that?

Finally, after review of this evidence, do you believe it makes more sense and is a simpler explanation that Baati et al have gotten the data of their study profoundly wrong with regard to Olive Oil than to take the CAIP as being important? Especially given the fact that Olive Oil has been shown to more powerfully trigger the CAIP than most (all?) other dietary substances in the gut?

With P < 0.05, there is only a barely significant correlation between HRV and surviving past 100. That says nothing about causality. The fact that HRV does an about face in late life doesn't mean it's causal of long life; it could just as easily mean that aging causes HRV. There is a lot of evidence that olive oil is good for you, and that the polyphenolic content is mainly responsible. (I'm personally a big fan, and have five liters of a great Piqual in the closet.) Still, I don't know of any evidence that olive oil alone can cause the sort of life extension reported by Baati. Given that, I'd rank the likelihood of explanation as Error > polyphenols > HRV. This could still mean that the combination of polyphenols and lipid-induced CAIP effects are at least parts of the explanation, assuming error is ruled out. To test if CAIP is possible as the primary explanation, rats could be dosed with olive oil that had been stripped of polyphenols.

[HighDesertWizard]

I agree. This Vagus stuff is very left field. In the meantime, we can rejoice that the two oldest people in the world until recently (both over 120) credited their longevity in part to olive oil.

et tu, Turnbuckle?

Does it feel "out of left field" because you've never heard of it? I completely understand that feeling. I felt the same way late last year. But then I actually looked at the studies. I recommend starting with the 2007 Review of the Literature... If you have looked at the studies, do you dispute their validity? If so, which one and how? Details...

We live in an age when more and more serious diseases that kill us off before 100 are understood, at least in part, to be Auto-Immune diseases. And the CAIP appears to be the key mechanism modulating the cytokine immune response... The Vagus-HRV-aging studies I've posted are consistent with the argument that individuals with modulated immune responses would experience, in general, more extreme longevity. How, specifically, is that argument coming out of left field?

I'm merely suggesting that Auto-Immune processes of the Vagus-CAIP might be important and that these processes might be important in explaining the Baati study result. I'm not saying I know. I'm merely saying it could be important. Experiments could determine if it is.

Final question... Is this "vagus stuff" more or less left field than niner's suggestion, playful? or serious?, just yesterday that Baati et al might have screwed up the Olive Oil data and other study data in some serious way?

When radical life extension is a reality, all the scientifically credible schools of thought about aging will be be able to point to actual evidence and say "see, I told ya so."

8-)