3585 replies to this topic

[Metrodorus]

Re fullerene supply: In the Ukraine, at least, fullerene in its hydrated form has been approved as a food additive and antioxidant for human consumption.

http://www.ipacom.co...ticles/fws1.pdf

[Junk Master]

Yup, that's the "watered down" stuff. Not the same at all.

[Metrodorus]

Experimental confirmation (2011) that fullerene is taken up and incorporated in cell membranes:
http://pubs.rsc.org/...1/CC/c1cc14650e

[Metrodorus]

Yup, that's the "watered down" stuff. Not the same at all.

The point of quoting that was to indicate that no matter what the FDA does,there will remain sources of supply for the pure reagent.

I cannot anyway imagine the pure reagent ever being controlled in any way whatsoever, as it is essentially non toxic, and it isn't a drug. Its uses are so multifarious in such a wide range of industries, that I cannot imagine a pathway that could lead to medical regulation of the raw product.

[smithx]

Experimental confirmation (2011) that fullerene is taken up and incorporated in cell membranes:
http://pubs.rsc.org/...1/CC/c1cc14650e

The picture on that link looks somewhat worrisome in that the position of enzymes within cell membranes is very important. They float at a certain depth within the membrane based on the location of hydrophilic and hydrophobic residues.

If C60 is in there changing the configuration of the membrane, it could affect the activity of these enzymes and cause all sorts of problems.

I hope it's a magic elixer, but it has lots of potential to do all kinds of other stuff.

[HighDesertWizard]

Experimental confirmation (2011) that fullerene is taken up and incorporated in cell membranes:
http://pubs.rsc.org/...1/CC/c1cc14650e

The point is, if C60 is a radical sponge, then even if it does catalyze some reactions, it will be changed by others and will get used up like other antioxidants get used up.

But that will be a slow process if there are no carbon radicals present, and these would be exceedingly rare species in vivo. I would expect that over a span of months or years, the C60 would essentially become 'used up', but I don't think it's going to happen on a rapid timescale. [Emphasis added]

It's still a mystery as to exactly how persistent the C60 is in the mitochondrial membrane. If the radionucleide C14 had a half life of 10-14 days in previous tests where it was being recycled in the new mitochondria being formed, I'd hazard a guess that the C60 might last a similar amount of time. It might be prudent to top off with an new dose every 10-14 days indefinitely. [Emphasis added]

From the peanut gallery, so far, I am behind Niner's theory of enhanced membrane lipid functionality. It gets cut the least by occam's razor. [Emphasis added]

I'm very confused about the status of the "C60-in-the-cell-membrane-is-the-essential-mechanism" argument vis-a-vis the Baati study life extension result.

According to the study, "pharmacokinetic studies show that dissolved C60 is absorbed by the gastro-intestinal tract and eliminated in a few tens of hours." Still, in various posts to this thread, the proponents of the "C60-in-the-membrane" argument have used various phrases to ascribe a much longer period of persistence of the C60 in the membrane.

Some questions...

• If it does turn out that C60 is cleared quickly, what's the "Plan B Backup Hypothesis" to support the "C60-in-the-membrane-as-SOD-mimetic-provdes-long-term-effects" argument? Is there a one? Does the entire argument hinge on the Baati team getting the clearance data wrong?

• If you make posts promoting the C60 in the membrane case without a Plan B Hypothesis about persistence duration, shouldn't you also be adding a footnote to those posts, at least once in a while, that says: "BTW, this argument assumes that Baati et al got the C60 clearance data wrong"...

• And Mind... How is believing that (1) Baata got the C60 clearance data wrong but (2) got the life extension result right, consistent with the simplicity principle of Occam's Razor?

Like I said, I'm confused. If I've misunderstood or misconstrued this essential question, please help me out and explain it to me.

Edited by wccaguy, 18 June 2012 - 10:58 AM.

[taho]

I think it goes something like this:

One part of C60/OO gets cleared from the blood in couple of days, but the other part of c60/OO gets integrated into cell membranes. If the cell would die and it's cell membrane would desintigrate, C60's hidrofobic nature would most likely mean that it would reintegrate itself in the first lipid membrane it would get in touch with. That means that C60 could stay in the body for a very long time and it wouldn't show up on any test in blood, urine or stool, since that part of c60 is not moving around the body in testables quantities.

By being in the middle of membrane, being catalist for neutralization of ROS, it can stop lipid peroxidiation chain reaction and stop further ROS from being formed.

Less lipid peroxidation, ROS formation => less damage to the body => better is body able to maintain itself => longer, healthier lifespan.

Edited by taho, 18 June 2012 - 12:59 PM.

[HighDesertWizard]

One part of C60/OO gets cleared from the blood in couple of days, but the other part of c60/OO gets integrated into cell membranes. If the cell would die and it's cell membrane would desintigrate, C60's hidrofobic nature would most likely mean that it would reintegrate itself in the first lipid membrane it would get in touch with. That means that could stay in the body for a very long time and it wouldn't show up on any test in blood, urine or stool, since that part of c60 is not moving around the body in testables quantities.

Ok. Are you saying the study authors were not familiar with the limitations of the approach they used to assess C60 clearance?

Are you saying the "pharmacokinetic studies" they performed are not adequate to the task of detecting all the C60/OO molecules, hence, the statement they make in the study abstract about clearance in "tens of hours" is technically incorrect, at best, and misleading, at worst?

Those are the implications of what you're saying, right? Isn't this a very serious objection to the basic scientific competence of the study authors?

[Turnbuckle]

Yup, that's the "watered down" stuff. Not the same at all.

The point of quoting that was to indicate that no matter what the FDA does,there will remain sources of supply for the pure reagent.

I cannot anyway imagine the pure reagent ever being controlled in any way whatsoever, as it is essentially non toxic, and it isn't a drug. Its uses are so multifarious in such a wide range of industries, that I cannot imagine a pathway that could lead to medical regulation of the raw product.

If the federal government wants to control or ban any product, they can do it under the commerce clause of the constitution. They could regulate quilting bees if they wanted to, according to Clarence Thomas in his dissent to a 2005 ruling. As for regulating pure reagents, that can be done under the precursor laws, which is any chemical reactant that takes part at any stage in the production by whatever method of a toxic chemical. So find that any C60 derivative is toxic and C60 could be regulated as a precursor.

[HighDesertWizard]

One part of C60/OO gets cleared from the blood in couple of days, but the other part of c60/OO gets integrated into cell membranes. If the cell would die and it's cell membrane would desintigrate, C60's hidrofobic nature would most likely mean that it would reintegrate itself in the first lipid membrane it would get in touch with. That means that could stay in the body for a very long time and it wouldn't show up on any test in blood, urine or stool, since that part of c60 is not moving around the body in testables quantities.

And I note that you didn't answer the other question I asked, namely, whether there is a Plan B Hypothesis just in case the Baati team aren't shown to be scientifically incompetent or imprecise in writing the study abstract and the C60 really did clear in "tens of hours."

Who has the answer to that question?

Edited by wccaguy, 18 June 2012 - 01:27 PM.

[taho]

Are you saying the "pharmacokinetic studies" they performed are not adequate to the task of detecting all the C60/OO molecules, hence, the statement they make in the study abstract about clearance in "tens of hours" is technically incorrect, at best, and misleading, at worst?

I am saying that we should read what was tested and what is writen.

The elimination half-lives indicate that C60 is completely eliminated from blood 97 h after administration irrespective of the route of administration.

Even if very small amount of c60 ends up in lipid membrane it is very powerfull antioxidant that would be just in a right place to stop lipid peroxidation.

That could even explain why Olive Oil alone was somehow better then control, since its mostly made of monounsaturated lipids, that are more resistant to oxidation then poliunsaturated lipids. If every mouse in the study was fed diet that has poliunsatured fats in them (seeds basicly), but "Olive Oil only" had extra monounstaturated fats in their. They could end up with less oxidative stress from less oxidative prone fats, thus live longer then controls.

Edited by taho, 18 June 2012 - 01:51 PM.

[Turnbuckle]

Even if very small amount of c60 ends up in lipid membrane it is very powerfull antioxidant that would be just in a right place to stop lipid peroxidation.

You see, wccaguy, the answer to your question can be found in homepathathy.

[arska]

Is it possible that C60 functions as liposomal delivery method for OO's tyrosol and hence has greater impact on SIRT1 and FOXO3?
This could contribute to explain why there is an observed effect even if the C60 is excreted.

[taho]

Even if very small amount of c60 ends up in lipid membrane it is very powerfull antioxidant that would be just in a right place to stop lipid peroxidation.

You see, wccaguy, the answer to your question can be found in homepathathy.

nice joke, but I am serious.

Pathological examinations show that even at very low doses,
500 times lower than that used previously [21], C60-olive oil solutions
effectively protects the livers against CCl4 toxicity. These
results are in agreement with those reported for very low doses of
water solution of hydrated C60 fullerene in other experimental
models

Rats were injected/feed large amounts of c60/OO. (they were testing for toxicity after all). It was in the blood for few days, then got either integrated in the tissue or excreted.

That means, if less then 100% of c60 is excreted from the body it is in the body somewhere. And since it even crosses the blood-brain barier it probably is everywhere. Even if 0,2% of that dose would stay in the body, that would be enough c60 to work it's magick for a long time.

Did 100% of c60 get excreted? I guess, no.

[HighDesertWizard]

Even if very small amount of c60 ends up in lipid membrane it is very powerfull antioxidant that would be just in a right place to stop lipid peroxidation.

You see, wccaguy, the answer to your question can be found in homepathathy.

Yes, Turnbuckle. One of my favorite scientists actually says this outright...

As I understand it, the claim is that the less you use Homeopathy, the better it works. Sounds plausible to me. -- David Deutsch



http://193.189.74.53...=20040808030513

Edited by wccaguy, 18 June 2012 - 03:03 PM.

[Turnbuckle]

The major objection to the epigenetic theory of action is that there is no evidence that fullerenes could act epigenetically. Except for this one--SiO(2) nanoparticles induce global genomic hypomethylation in HaCaT cells*, which suggests that nanoparticles can do this even though they aren't designed to do anything in particular. It's a function of size, apparently. And there's this paper--Epigenetic modulation of human breast cancer by metallofullerenol nanoparticles. These particles were C82 with a single metal adduct. So still not there, but getting closer.

*I wonder if this might be the mechanism of "Microhydrin," a nano-silica product that has been sold for some time as a super antioxidant. The stuff was invented by a pyramid power promoter and the marketing is a total scam, but it apparently has some real effects.

[NewtonPulsifer]

One part of C60/OO gets cleared from the blood in couple of days, but the other part of c60/OO gets integrated into cell membranes. If the cell would die and it's cell membrane would desintigrate, C60's hidrofobic nature would most likely mean that it would reintegrate itself in the first lipid membrane it would get in touch with. That means that could stay in the body for a very long time and it wouldn't show up on any test in blood, urine or stool, since that part of c60 is not moving around the body in testables quantities.

And I note that you didn't answer the other question I asked, namely, whether there is a Plan B Hypothesis just in case the Baati team aren't shown to be scientifically incompetent or imprecise in writing the study abstract and the C60 really did clear in "tens of hours."

Who has the answer to that question?

It is a very good question. I did a quick google of "C14 labelled fullerene" and the first hit was:
In vivo biological behavior of a water-miscible fullerene: ¹⁴C labeling, absorption, distribution, excretion and acute toxicity

"When administered orally to rats, this compound was not efficiently absorbed and was excreted primarily in the feces. When injected intravenouesly, however, it was distributed rapidly to various tissues, and most of the material was retained in the body after one week."
http://www.ncbi.nlm..../pubmed/9383440

Is this directly relevant to lipofullerene adducts? No....but it does establish there are fullerene varieties that are biologically persistent. I don't have time at the moment but I'll research this more.

We wouldn't really have the proof until someone does an experiment with the C60 adducts carefully separated and individually tested (is it C60/Oleic Acid, C60/Alpha-Linoleic Acid, C60/Chlorophyll, a combo?), and then also carefully creates a C14 labelled lipofullerene to get data on bio-accumulation.

Edited by NewtonPulsifer, 18 June 2012 - 04:07 PM.

[taho]

One part of C60/OO gets cleared from the blood in couple of days, but the other part of c60/OO gets integrated into cell membranes. If the cell would die and it's cell membrane would desintigrate, C60's hidrofobic nature would most likely mean that it would reintegrate itself in the first lipid membrane it would get in touch with. That means that could stay in the body for a very long time and it wouldn't show up on any test in blood, urine or stool, since that part of c60 is not moving around the body in testables quantities.

And I note that you didn't answer the other question I asked, namely, whether there is a Plan B Hypothesis just in case the Baati team aren't shown to be scientifically incompetent or imprecise in writing the study abstract and the C60 really did clear in "tens of hours."

Who has the answer to that question?

I checked the paper for an answer and have found this nugget of information:

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 [21] as it has been
recently confirmed [25]. 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 [40]. 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 [21,42]. 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.

So.. some of it actually does not get excreted, and stays in the body.

If it is incorporated into cellular lipid walls, that would mean that it would not only stay there, but it would be a low quantity reservoir of c60, which would be released by apoptosis and recycling of material in new cells. That would explain longevitiy efect, even after they stoped giving c60/OO to rats when the controls died.

I read somewhere they died of "old age" instead of the usual "tumors and lung failures", so my guess is c60 worked till the end and other things unrelated to oxidative stress killed the animals (cell loss, cell athropy, death resistant cells, AGEs, extracellular and intracellular aggregates - take a pick).

Edited by taho, 18 June 2012 - 05:09 PM.

[Turnbuckle]

...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.

So.. some of it actually does not get excreted, and stays in the body.

.

That doesn't follow from the quote.

Edited by Turnbuckle, 18 June 2012 - 05:07 PM.

[NewtonPulsifer]

Ok. Are you saying the study authors were not familiar with the limitations of the approach they used to assess C60 clearance?

Are you saying the "pharmacokinetic studies" they performed are not adequate to the task of detecting all the C60/OO molecules, hence, the statement they make in the study abstract about clearance in "tens of hours" is technically incorrect, at best, and misleading, at worst?

Those are the implications of what you're saying, right? Isn't this a very serious objection to the basic scientific competence of the study authors?

The pharmacokinetic analysis is page 3, section 2.7 of the paper. It only references plasma concentration.

So the reference to clearance in "tens of hours" is in reference to blood plasma clearance *only*.

[maxwatt]

C60 may be cleared from blood in a matter of hours after being absorbed by the gut, but did they test where it went? I believe none was found in urine, I don't know if they measured in feces. But being in the cell membrane would not count as being in the blood. They tested by drawing blood samples. So the clearance data is not inconsistent, depending on one's method of measuring and defining clearance.

Having an hypothesis to test is enough; a backup hypothesis is not necessary. If it turns out C60 is not present in the cell membrane following in vivo administration, then it's up to the "theorists" to come up with an alternate hypothesis to test. The cell-membrane hypothesis is consistent with what is currently known, and dovetails nicely with the "ROS free radical damage to mitochondria" theory of aging. We don't need an alternate theory to test it. If and when it is established that C60 is present and possibly acting in the cell membrane, then we can figure out a way to measure what it is doing there.

[taho]

That doesn't follow from the quote.

Hmm.. I think you are right.. It is circumstantial at best. But you can look at it in the other way..

If and when it is established that C60 is present and possibly acting in the cell membrane, then we can figure out a way to measure what it is doing there.

In the paper they did disect rats and find c60 in blood, liver, spleen and brains of those rats. They could see C60 crystals containing macrophages with specific brown colour. What happens to those crystals? This?

Our aim was to establish if human monocyte macrophages internalise nanoparticles and to assess whether nanoparticles are modified by cells following uptake. Using these techniques we were able to show a marked increased in the amount of information gained from 3-D imaging. 3-D electron tomography revealed several sub-cellular compartments containing C(60) within the cell: secondary lysosomes, along the outer and nuclear membrane and most notably inside the nucleus of the cell. Using EFTEM and STEM-based techniques we were able to visualize cell structures such as membranes, the mitochondria, ribosomes and the nucleus, without the need for traditional staining techniques.

http://www.ncbi.nlm....pubmed/16765881

So.. they get there.. how long do they stay in the walls?

Edited by taho, 18 June 2012 - 06:05 PM.

[JohnD60]

Sarah, as far as recouping your investment, I'd focus your marketing on non-longevity related benefits. Turnbuckle is exactly right when he says, "Hey, isn't it enough that it gives you good hair," because good hair would be enough for a billion dollar market worldwide, as evidenced by Rogaine's 700 million dollar U.S. sales.

Again, if the product produced even a 5% increase in Vo2Max it's use would sweep the sporting/fitness industry and would also be a multi-billion dollar market.

I wonder if there is a new member researching the VO2Max angle named Juan Pelota (Lance Armstrong's alias).
As far as the reported hair growth, I am skeptical, primarily because I know how slow hair grows, about .5"/month, and that it grow in waves (I have had Hair transplants in the past, am a former finasteride user and a current minoxidil user), it doesn't seem to me as if there has been enough time since the study was reported to have a noticeable increase in hair growth for anyone

[Metrodorus]

Re Bioaccumulation - the particular fullerene adduct is very important here.

There is wide variation in bioaccumulation reported in the literature, depending on what adducts are present.

As we do not know what the adducts are in fullerne-olive oil, we cannot say much.

Clearance from the blood is not a useful measure of bioaccumulation.
If the fullerene adducts generated in olive oil accumulate in the liver, then they may be redistributed from there to other organs.

A good summary of the extant literature ( up to 2008 ) on bioaccumulation can be found here: under the heading "absorption, distribution and elimination", about half way through the paper.

The key paragraph is this one:
" Excretion may be absent, a very slow process or a relatively fast process, which was the case with a fullerol and a C₆₀-PEG derivative. Thus, bioaccumulation can be prevented by an appropriate selection of derivatives"

[Metrodorus]

This study is important:

http://www.sciencedi...958166911006136

It points out that fullerene aggregates are not toxic as originally thought, and observed toxic effects were the result of experimental artefacts in the early studies.

Thus, non-filtration / centrifuging of the olive oil - fullerene mix might not be as risky as originally surmised.

"This review aims to clarify inconsistencies in previous reports regarding the potential for aqueous aggregates of fullerenes (nC₆₀) to generate reactive oxygen species (ROS) and cause toxicity in fish. Methods for evaluation of ROS production and toxicity of aqueous nC₆₀ have evolved over time and limitations in initial studies have led to unintentional erroneous reports of nC₆₀ ROS generation and toxicity. Some of these reports continue to lead to misconceptions of the environmental effects of C₆₀. Critical review of the evidence (2007–2011) indicates that aqueous nC₆₀ have minimal potential to produce ROS and that oxidative stress in fish is not induced by environmentally relevant exposure to nC₆₀. Future studies should acknowledge that current evidence indicates low toxicity of nC₆₀ and refrain from citing articles that attribute toxicity in fish to nC₆₀ based on methods shown to be compromised by experimental artifacts. "

Edited by Metrodorus, 18 June 2012 - 07:11 PM.

[Metrodorus]

Bioretention of a metallo-fullerene gives some idea of what might happen......with the usual caveats.

This is a totally different molecule to our olive oil fullerene adducts - so probably not much light is thrown by these results.


http://www.pnas.org/.../96/9/5182.full

Edited by Metrodorus, 18 June 2012 - 07:17 PM.

[Turnbuckle]

Bioretention of a metallo-fullerene gives some idea of what might happen......with the usual caveats.

This is a totally different molecule to our olive oil fullerene adducts - so probably not much light is thrown by these results.


http://www.pnas.org/.../96/9/5182.full

That's a plot of the control compound, not the C60.

[Metrodorus]

oops.

This (2010) toxicology review of various fullerene adducts is interesting and thorough

http://toxsci.oxford.../114/2/162.full

[HighDesertWizard]

Having an hypothesis to test is enough; a backup hypothesis is not necessary. If it turns out C60 is not present in the cell membrane following in vivo administration, then it's up to the "theorists" to come up with an alternate hypothesis to test. The cell-membrane hypothesis is consistent with what is currently known, and dovetails nicely with the "ROS free radical damage to mitochondria" theory of aging. We don't need an alternate theory to test it. If and when it is established that C60 is present and possibly acting in the cell membrane, then we can figure out a way to measure what it is doing there.

I now think the statement in the Baati study abstract is misleading and it's reasonable to assume C60 adducts for some period. The clearance issue was my major concern about the "C60-in-the-cell-membrane" hypothesis. I'm glad I pressed the point for several reasons, I think Turnbuckle's argument up thread against longer persistence is the best one I've seen... 'nuff from me about this...

That said, I believe the "Cell-Membrane-ROS-Scavenging-As-Essential Mechanism-Explanation" folks should have have a backup hypothesis. The longer term life span enhancing effects experienced by the Baati rats have to be explained somehow. Earlier up thread, niner had tied these effects specifically to the C60 extended stay in the membrane hypothesis.

... 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 may be right. But in case he's wrong, how does the SOD mimetic/radical scavenging as Essential Mechanism Explanation stay alive? That is why I pressed the point this morning. It needs to be kept alive if C60 clears relatively quickly. We need more explanations, more testable hypotheses, not less... The more diverse and better ideas we generate for the theorists to consider the sooner we'll see the results of the tests we want to see. I propose that we give the theorists even more good reason to pay attention to this discussion...

Edited by wccaguy, 18 June 2012 - 08:44 PM.

[niner]

Is it possible that C60 functions as liposomal delivery method for OO's tyrosol and hence has greater impact on SIRT1 and FOXO3?
This could contribute to explain why there is an observed effect even if the C60 is excreted.

If you mean the tyrosol is inside the fullerene cage, then no, it's not possible. Small molecules like tyrosol can not get inside the fullerene cage. The conditions for liposome formation don't appear to exist in the oo/C60 mixture. Aggregates are filtered out, so if the hypothesis were that the tyrosol is sneaking in by hiding inside a very small aggregate that managed to pass through the filter, I don't think there could be enough mass of C60 in that form to bring in enough tyrosol to matter. Whether the C60 were excreted or not, the tyrosol would be excreted, since it would be rapidly conjugated by phase II xenobiotic metabolism, and would be quickly eliminated. This would also apply to any other phenolic compounds in the olive oil as well. In short, I don't think that EVOO phenols, chlorophyll, or any other non-lipid component of the OO has anything to do with the effect of the C60. On the other hand, rats that were fed EVOO by gavage, 24 times in the course of their lives were reported to live significantly longer than those given water. If we believe the C60 arm of the study, then we all ought to be swigging EVOO, since it's certainly on the GRAS list. It's funny how we're all ignoring that part of the study. If we don't believe it, then is there a reason to believe the C60 arm? If we do believe it, then we should start drinking EVOO. (I like EVOO, but 4/5 of a cup doesn't seem very appealing.)