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C60 experiments @ home

buckyball c60 fullerene buckyballs

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#661 Turnbuckle

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Posted 09 June 2012 - 08:18 PM

Slightly off topic, but for storage purposes, Miron glass seems better than amber bottles with regard to blocking UV:
http://www.idspackag...g_supplier.html
Just thought I would post this here also in case it is of interest to anyone.

Scratch that, Miron lets UVA in, looks like black glass would be best...


That miron glass looks almost black so it would be hard to tell what you had in there--a beautiful purple solution or a horribly rancid brown one. Better just to keep it in the fridge.

#662 AgeVivo

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Posted 09 June 2012 - 09:36 PM

Sorry to slow down the temperature for some, but the #1 rule in biology at this stage is that theories are wrong. Numerous papers are difficult to reproduce, in part because results may depend on some specific things that are present in a lab (eg cell dish in plastic A rather than plastic B) and no one even knows it is because of it. So, results from studies should be extrapolated a little rather than a lot. It is not like playing chess with thinking 6 moves in advance.

It doesn't mean that we should'nt think, in the contrary. The only thing is that we should understand that most often our thinkings, however nice they sound, will prove wrong. So either try to find theories that are only slight extrapolations of many papers, if possible, or have a lab where you test test test test things to progress one step at a time; try things and analyze as much as you can. It is generally the best way to go forward in biology. Most great discoveries are said to be luck, but if you analyze them they are actually 99.9% transpiration and 0.1% luck, the luck coming from numerous side testings and analysis rather than being purely driven by theory and standard protocols.

Edited by AgeVivo, 09 June 2012 - 09:42 PM.

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#663 smithx

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Posted 09 June 2012 - 09:50 PM

I don't see in that paper where hypomethylation of the mtDNA causes any specific problem.


Methylation is one of the methods used by the cell to control gene expression. It is not better if every gene is expressed to the same degree. This is basic biology. The point I was hoping that paper would make clearer is that methylation performs the same function for mitochondrial DNA as it does for nuclear DNA.

A non-specific demethylation agent would cause problems by increasing expression of genes which should be suppressed. See these or other similar papers:
https://docs.google....WUSg0WjiAKp47Kw

http://www.biomedcen...1741-7015/10/26


So C60, if it is beneficial, is very unlikely to be a non-specific demethylation agent.

Since there's no evidence at all that C60 has anything to do with methylation, and since non-specific demethylation agents would be harmful, I would hope this wild speculation about methylation could be put to rest.

Edited by smithx, 09 June 2012 - 09:53 PM.

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#664 revenant

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Posted 09 June 2012 - 09:58 PM

C60 has been observed tenuously "encapsulating" patassium ions (up to four at a time), and is also know to disrupt hydrogen bonds. It would not surprise me if C60 had the ability to demethylate CH3+ cations from DNA.

Edited by revenant, 09 June 2012 - 09:59 PM.


#665 Turnbuckle

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Posted 09 June 2012 - 10:07 PM

Methylation is one of the methods used by the cell to control gene expression. It is not better if every gene is expressed to the same degree. This is basic biology. The point I was hoping that paper would make clearer is that methylation performs the same function for mitochondrial DNA as it does for nuclear DNA.

A non-specific demethylation agent would cause problems by increasing expression of genes which should be suppressed. See these or other similar papers:
https://docs.google....WUSg0WjiAKp47Kw

http://www.biomedcen...1741-7015/10/26


So C60, if it is beneficial, is very unlikely to be a non-specific demethylation agent.

Since there's no evidence at all that C60 has anything to do with methylation, and since non-specific demethylation agents would be harmful, I would hope this wild speculation about methylation could be put to rest.


These papers don't address mtDNA and so don't make your point While it's true that methylation is important in distinguishing one cell type from another via the epigenome of the nuclear DNA, it does not appear so important with mitochondria. Different cell types have different numbers of mitochondria, but they all seem to be of one type. As for this "wild speculation," changes to mitochondrial methylation would be an easy explanation for how a few treatments with C60 could have such lasting effects. The epigenome is much more malleable than the genome, yet persistent enough to explain the results. Other explanations can't account for the persistence.

Edited by Turnbuckle, 09 June 2012 - 10:12 PM.


#666 smithx

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Posted 09 June 2012 - 10:29 PM

These papers don't address mtDNA and so don't make your point


OK how about this one:
https://digarchive.l...ndle/10156/3563

The point to take away from this article is that if mitochondrial DNA methyltransferase 1 is highly conserved, it must be vital for the function of the mitochondria.

This article shows that methylation of mitochondrial DNA varies by area, and again implies that this methylation is doing something important:
http://www.ncbi.nlm....pubmed/19740762

Methylation in the wrong places can certainly cause harm, but it's a part of a basic system used to control gene expression. It's certainly not true that all genes should be expressed as much as possible. Again, this is a basic fact of biology: each system is in a very precise state of balance. A big part of that is maintained by feedback mechanisms which control the degree of expression of genes, and a basic part of those mechanisms is methylation. Anything which disrupts those systems is likely to cause severe problems.

And again, there's zero evidence that C60 would have anything to do with this system.

#667 Turnbuckle

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Posted 09 June 2012 - 10:56 PM

OK how about this one:
https://digarchive.l...ndle/10156/3563

The point to take away from this article is that if mitochondrial DNA methyltransferase 1 is highly conserved, it must be vital for the function of the mitochondria.

This article shows that methylation of mitochondrial DNA varies by area, and again implies that this methylation is doing something important:
http://www.ncbi.nlm....pubmed/19740762

Methylation in the wrong places can certainly cause harm, but it's a part of a basic system used to control gene expression. It's certainly not true that all genes should be expressed as much as possible. Again, this is a basic fact of biology: each system is in a very precise state of balance. A big part of that is maintained by feedback mechanisms which control the degree of expression of genes, and a basic part of those mechanisms is methylation. Anything which disrupts those systems is likely to cause severe problems.

And again, there's zero evidence that C60 would have anything to do with this system.


There's not zero evidence. There's theoretical work that suggests C60 binds to DNA, and C60 is certainly large enough to interfere with the action of methyltransferase. The paper you linked to says, "We propose that the enzymes responsible for epigenetic modification of mtDNA have potential as therapeutic targets, with relevance to a broad spectrum of human disorders," and that is perhaps what C60 is doing if it reversibly binds to mtDNA, thus acting as a methyltransferase inhibitor. Another methyltransferase inhibitor, lidocaine, is also believed to bind to mtDNA and has been shown to extend the lifespans of Wistar rats.

#668 Mind

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Posted 09 June 2012 - 11:53 PM

Just wanted to chime in on the speculative parts of this thread. With this study, there is only theory and speculation as to the method of action. Thus many possibilities are open. Hopefully we can close some branches of speculation by conducting our own study, please contribute your thoughts here for potential new research.

Also, please let me express my sincerest gratitude to everyone for keeping this very informative and interesting discussion - civil. I am certain it is one of the qualities that has made this thread one of the hottest life extension discussions on the net in the last few weeks.

There are many different theories bubbling up that need to be rationally analyzed. A few times in the past some very intriguing discussions devolved into flame wars. Not here. Let's keep the discussion rolling!
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#669 JohnD60

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Posted 10 June 2012 - 02:22 AM

First, we need to characterise the molecules that form in the olive oil....


I will take that as an invitation to post my most recent completely speculative theory about the interaction of Olive Oil with C60. :) My hypothesis is that the polyphenols in the OO wrap themselves around the C60 and hydrogen bonds form linking the two together. This hypothetical linkage could create two very different possible beneficial results... 1. The polyphenol shield protects the C60 from biochemical attack, and allows the C60 to pass through the digestive process without being broken down. 2. The hydrogen bonds serve to provide the polyphenols with increased resistance to chemical attack, and allows the polyphenols to pass through the digestive process without being broken down.

#670 niner

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Posted 10 June 2012 - 02:46 AM

If you only kinda know how a car engine works, and your car needed a new timing chain, would you fix it yourself? No, you'd have a mechanic do it. If the main electrical service panel at your house shorted out, and you aren't an electrician, would you rewire it yourself? No, you'd hire someone qualified to do it. Now, considering that life is the most complex chemico-physical phenmonon in the known universe, everyone should ask themselves: "Am I qualified to propose mechanistic explanations for the apparent results of an unusual pharmacologic intervention?"

#671 niner

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Posted 10 June 2012 - 02:52 AM

C60 has been observed tenuously "encapsulating" patassium ions (up to four at a time), and is also know to disrupt hydrogen bonds. It would not surprise me if C60 had the ability to demethylate CH3+ cations from DNA.

It would surprise the hell out of me. Do you know how high energy a CH3+ cation is?

#672 niner

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Posted 10 June 2012 - 03:09 AM

As for this "wild speculation," changes to mitochondrial methylation would be an easy explanation for how a few treatments with C60 could have such lasting effects. The epigenome is much more malleable than the genome, yet persistent enough to explain the results. Other explanations can't account for the persistence.


What about the explanation that the fullerene-fatty acid adducts populate the membrane compartment, and remain there for a year or more? That would explain it. Before someone says that they clear in ten hours (or whatever it was), we first need to know what 'clearance' means in a pharmacokinetic experiment. After dosing the animal, blood is periodically withdrawn from an indwelling catheter. The blood will be extracted with solvent and analyzed by HPLC. When the analyte peak is within the noise in the chromatogram, it has "fully" cleared to most people's satisfaction. One of the places the C60 may be clearing TO is the membrane compartment. Membranes are made of fatty acids, so fatty acids like to hang out in membranes. They like to be with their own. If they happen to have a C60 attached covalently, it will come along for the ride.
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#673 niner

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Posted 10 June 2012 - 03:20 AM

There's not zero evidence. There's theoretical work that suggests C60 binds to DNA, and C60 is certainly large enough to interfere with the action of methyltransferase. The paper you linked to says, "We propose that the enzymes responsible for epigenetic modification of mtDNA have potential as therapeutic targets, with relevance to a broad spectrum of human disorders," and that is perhaps what C60 is doing if it reversibly binds to mtDNA, thus acting as a methyltransferase inhibitor. Another methyltransferase inhibitor, lidocaine, is also believed to bind to mtDNA and has been shown to extend the lifespans of Wistar rats.


Molecular dynamics simulation is a step up from zero, just not a very large step. However, this idea that C60 sits in the groove and interferes with methyltransferase makes no sense at all. If that's the case, how would the methyltransferase do its job? If you're postulating either enhanced methylation or demethylation, then interfering with methyltransferase isn't what you want to do. The C60 getting in the way would result in no change to methylation.

How does your hypothesis work if the active species is a C60 substituted with one or two fatty acids?

#674 Mind

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Posted 10 June 2012 - 12:10 PM

A couple previous discussion about cell membrane fatty acid composition and life extension, here and here.

#675 Metrodorus

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Posted 10 June 2012 - 02:44 PM

Another piece in the puzzle of fullerene in lipid activity:

http://www.springerl...812275587w8855/
Summary:
Overall C60 appears to play an antioxidant role that is modulated by Polyunsaturated Fatty Acids (PUFA), taking into account its effects on intracellular ROS concentration and MDA levels. Results also suggest that C60 influences GSH synthesis, as showed for the augmented levels of this antioxidant and also for the lowering of the intracellular cysteine concentration.

#676 Turnbuckle

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Posted 10 June 2012 - 03:03 PM

Molecular dynamics simulation is a step up from zero, just not a very large step. However, this idea that C60 sits in the groove and interferes with methyltransferase makes no sense at all. If that's the case, how would the methyltransferase do its job? If you're postulating either enhanced methylation or demethylation, then interfering with methyltransferase isn't what you want to do. The C60 getting in the way would result in no change to methylation.

How does your hypothesis work if the active species is a C60 substituted with one or two fatty acids?


Why does it make no sense? The diameter of DNA is about 20A and the diameter of C60 is about 10A. Even without doing any computer analysis it would seem geometrically favorable for the spherical C60 to sit in the groove of the helix where it has two points of attraction rather than just one. And even without considering the structure of methyltransferase, it would seem impossible for it to function without communicating across the groove and thus C60 would act as a physical barrier. During replication in the presence of C60, two daughter loops are formed that are hemimethylated to some degree because C60 is suppressing methylation of the newly constructed strands. And during the next generation, if still in the presence of C60, half of the granddaughter loops will now lack any methylation where the daughter loops were hemimethylated. So the C60 would have to be present for at least two generations for its effects to become permanent. This might be as little as two days for more actively dividing mitochondria.

Edited by Turnbuckle, 10 June 2012 - 03:08 PM.


#677 Metrodorus

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Posted 10 June 2012 - 03:13 PM

Another line of research on which there are already a number of papers, covers the issue of where the fullerenes end up in the cell membranes.

It appears that the fullerenes are readily drawn into the membrane. (2012 theoretical study here: http://www.springerl...5p784863284710/ )

It also appears that they do not penetrate, but end up congregating in the hydrophobic interior of the membrane.
(2011 study here: http://pubs.rsc.org/...1/cc/c1cc14650e )

Posted Image

Some off the cuff questions that arise:
a. What effect do the fullerenes have on overall molecular stability of the membrane, and its longevity? I would imagine a direct overall organism level longevity effect if membranes were protected from oxidation by incorporation of fullerenes.

Long term incorporation into membranes could explain the persistent effects observed.

b. How persistent is the fullerene? If a cell undergoes apoptosis, is the fullerene drawn into the membranes of surrounding cells?

c. What happens to fullerene that enters the cytoplasm? It it incorporated into organelle membranes too, apart from the mitochondria?

d. How is fullerene incorporation into mitochondrial lipid membranes characterised?

Edited by Metrodorus, 10 June 2012 - 03:14 PM.


#678 revenant

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Posted 10 June 2012 - 03:37 PM

It would surprise the hell out of me. Do you know how high energy a CH3+ cation is?


Though I do anyway, I am not qualified to speculate about chemical physics (I mispell potassium :laugh:). Based the conjecture of the computer modeling study that proposed C60 (though not C60/oo) had the ability to split the double strand at the hydrogen bonds between bases, is it counter intuitive to think C60 could cause dissociation of hydrogen bonds in the methyl group?

#679 Metrodorus

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Posted 10 June 2012 - 03:39 PM

With all the usual caveats, this study on the protective effects of fullenerol points to some possible modes of action:
(See paragraph 5 )


Many studies have been carried out on fullerene and mineral oils, for much longer than fullerene and organic oils such as olive oil.

Some of the research results may possibly shed some light....although I admit this is unlikely.

http://www.ipme.ru/i.../ginzburgEN.htm

Incorporation of fullerene into the lipid membrane would change membrane chemistry.

Could some type of fullerene polymerisation be taking place at a membrane level as well, following incorporation?

Is fullerene polymerisation taking place with the fullerne-olive oil solution prior to ingestion?

Edited by Metrodorus, 10 June 2012 - 04:06 PM.


#680 Metrodorus

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Posted 10 June 2012 - 04:43 PM

Food for thought?
Posted Image

http://acswebcontent...PORTS/P7877.HTM

#681 smithx

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Posted 10 June 2012 - 05:36 PM

Why does it make no sense? The diameter of DNA is about 20A and the diameter of C60 is about 10A. Even without doing any computer analysis it would seem geometrically favorable for the spherical C60 to sit in the groove of the helix where it has two points of attraction rather than just one. And even without considering the structure of methyltransferase, it would seem impossible for it to function without communicating across the groove and thus C60 would act as a physical barrier.


If there was a molecule strongly bound to DNA, blocking enzyme activity, it would block transcription as well. Cells would not be able to replicate and genes would not be able to be expressed. The cell would die.

I hope it is clear that it's not possible to postulate that C60 binds just strongly enough with DNA to prevent methylation, while not interfering with any other enzymes.

Edited by smithx, 10 June 2012 - 05:39 PM.


#682 Turnbuckle

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Posted 10 June 2012 - 05:48 PM

If there was a molecule strongly bound to DNA, blocking enzyme activity, it would block transcription as well. Cells would not be able to replicate and genes would not be able to be expressed. The cell would die.

I hope it is clear that it's not possible to postulate that C60 binds just strongly enough with DNA to prevent methylation, while not interfering with any other enzymes.


It might very well decrease transcription, just as lidocaine does.

#683 niner

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Posted 10 June 2012 - 06:16 PM

Based the conjecture of the computer modeling study that proposed C60 (though not C60/oo) had the ability to split the double strand at the hydrogen bonds between bases, is it counter intuitive to think C60 could cause dissociation of hydrogen bonds in the methyl group?


There's been some misunderstanding in this thread about what exactly a hydrogen bond is. Classic hydrogen bonds only occur when there is a hydrogen covalently attached to an electronegative element like oxygen or nitrogen. The electrons in this bond are attracted to the more electronegative element, resulting in a partial positive charge on the hydrogen and a partial negative charge on the oxygen or nitrogen. Thus you have a small electrostatic dipole. This dipole can interact electrostatically with others like itself, as such:

O-H - - - - O-H


In this example, the dashed line is the hydrogen bond, and the other bonds are normal covalent bonds. Here's the important thing: The energy needed to break a hydrogen bond is low; between 5-30kj/m. The energy needed to break a covalent bond is large, like 350kj/m for a carbon-carbon single bond.

Hydrogen bonds are stronger than Van der Waals interactions, but covalent bonds are one to two orders of magnitude stronger.

Pristine C60 has no electronegative atoms, thus can not engage in hydrogen bonds as they're normally defined.

#684 Metrodorus

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Posted 10 June 2012 - 07:23 PM

Chemistry of fullerene in solution:
https://intranet.nas...nsformation.pdf

#685 HighDesertWizard

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Posted 10 June 2012 - 08:47 PM

wccaguy - science isn't philosophy - not since Galileo anyway.

You keep persisting in positing these modes of action, when we don't even know what molecule(s) we are dealing with!!!!!

It is all, quite frankly, rather far fetched.

First, we need to characterise the molecules that form in the olive oil.

I assume there are several reaction products.

Then, the metabolic effect of these need to be teased out, and that will take years, as if there are several reaction products, there will be several variables.

If the molecules work synergetically, then it will be even more difficult to untangle, as designing experiments that deal with multiple variables is fraught with difficulty.

The mechanisms of action posited so far in the extant literature are sufficient to explain what Mr Turnbuckle has felt - simply increasing the efficiency of the electron transport chain in the mitochondria will account for that. And we already know that fullerene in some of its forms has this effect. We do not need to posit any epigenetic changes. We do not even have to posit a direct effect on electron transfer, simply physical blockage of reactive species from access.

At this point we do not actually need any explanations. We simply need the original experiment to be reproduced, preferably more than once. Once that has been done, we will see whether the result was an artefact or not.


Turnbuckle has provided evidence that he is a credible person. And so I believe his truly remarkable, if anecdotal, report. So, therefore, I take it that it's very likely, though not certain, that something remarkable happened with the Baati rats as well.

That's it, it's that simple. Who disagrees?

Metrodorus... That said, we have a difference of opinion about how scientific progress takes place. Call it a "philosophical" difference if you like.

You argue that we cannot quicken the pace of discovery through Conscious and Deliberate Explanation Generation because we don't know enough about Fullerenes. More than that, you argue that we OUGHT NOT proceed with Explanation Generation until the Science of Fullerenes is understood, either completely or close to completely: "More must be proven about Fullerene Science before we can know anything...Stop Generating Explanations..."

I couldn't disagree more. The fact is, the subject matter that Explanations have to be built around are the sciences of Human Biology and Physiology and those sciences, however new, have a voluminous literature. We know a lot about ourselves and how and why we age. And the science of Fullerenes does not make settled science about homo sapien Biology/Physiology mysteriously disappear.

At this stage, I believe it's a huge mistake to concern ourselves with Proving anything. I believe we need to be generating More Explanations, not less, that (1) generally fit existing evidence, (2) are Falsifiable, and (3) Hard to Vary.

Metrodorus... Your argument that we shouldn't be generating Explanations isn't really with me, it's with David Deutsch, the noted Oxford Physicist. I've posted an 18 minute TED Talk Dr. Deutsch gave in 2009 about The Nature of Scientific Theory and the Reason why progress has exploded since the Enlightment. I recommend watching it. Let's not get this thread off track. I'd be happy to discuss or debate Dr. Deutsch's (and Karl Popper's) ideas with you in that thread if you like...

Metrodorus... It's clear you have serious knowledge about Fullerene Science. Can we count on you to help us brainstorm the experiments we might do to Replicate the C60/Olive Oil Study? Mind has now pointed to this thread twice, asking folks to participate to move the project definition forward. I've posted version 0.001 of a matrix to organize the kinds of experiments that should be done. That draft needs addition, enhancement, and deletions. Knowledge of Fullerene Science is important to making progress. Can we count on you to help so your knowledge begins to make itself felt by version 0.003?

More Explanations, not fewer, plausibly fitting existing evidence, are the foundation for Generating more Hypotheses that are Falsifiable and Hard to Vary. Generating these Explanations, and the Falsiable Hypotheses they imply, is the path to more and faster progress... Who's In?

#686 HighDesertWizard

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Posted 10 June 2012 - 09:16 PM

Turnbuckle, niner, et al, anyone, everyone... The discussion is amazing and we all hope it continues.

It would be great, also, if you could think about what experiments might be performed to Falsify your own hypothesis or the hypothesis of someone else in the discussion. I might be wrong, but it seems to me we could test some of the hypotheses floating around without doing full-scale life span studies in animals. Right?

Post descriptions of experiments to Falsify your own or someone else's hypothesis in this Replicating the C60/Olive Oil Study forum thread. I'll make sure to include it soon as a potential project study in a Google document that you have accessibility to edit.

Edited by wccaguy, 10 June 2012 - 09:18 PM.


#687 Metrodorus

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Posted 10 June 2012 - 09:35 PM

wcca guy:
It is far too early to be extrapolating in as much detail as proposed.


We don't need explanations at this point, we need good, sharp questions.
Science advances by forming hypotheses, which we test. These are also based on conceptual frameworks, which may sometimes be less grounded. No argument there.

But to do this, certain base data is needed.

I suspect that we don't have firm facts from which to extrapolate in detail in a meaningful way.

We have a number of fullerene studies, all dealing with different adducts.

We know there are orders of magnitude differences in observed behaviour of the adducts. We cannot just talk about 'fullerene' as a single molecule. It isn't that simple.

Extrapolating from the results of a published fullerene study that is not dealing with our particular to-date uncharacterised fullerene adduct(s), is going to be fraught with problems.

We don't know what the adducts are that form in the olive oil.
We don't know which ones are the active constituent.
We don't know their bioreactivity.
We don't know where these particular fullerene adducts end up at a cellular level - in the mitochondria? Maybe.
In the nucleus? Maybe.
In the cell membranes? - Almost definitely.
In various organelles like the endoplasmic reticulum? Maybe.
Do we know what they do in any of these locations? No, we do not have a clue.
Do we know how persistent they are once absorbed? No.
Do we know how they are excreted? No.
Until we have some actual data, more detailed explanations are going to be tenuous.

I think we should free to continue to generate explanations - but be aware of the limitations.


Certainly, some posited interactions can be ruled out outright, or given a very low probability, for chemical / physical reasons / biological reasons.

To reiterate, wccaguy, I do agree that we should continue to posit mechanisms of action - and debate their plausibility - but with the caveat that we really are flying blind.
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#688 Anthony_Loera

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Posted 10 June 2012 - 10:39 PM

Well,

What I do know is that taking 4/5 cup of oil at night is much more disgusting than taking it in the morning.... And doesn't give ya night time indigestion.

:p hehehe

Cheers
A

#689 Allen Walters

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Posted 10 June 2012 - 10:55 PM

Well, I got the replacement 99.5% c60 from Solaris chem inc. The product code on this is SOL5060X, and it's a black powder with no smell. I put .18 gr in 200 ml of the Carapelli first cold pressing extra virgin olive oil that I purchased from walmart. I put it on the magnetic mixer at 5pm yesterday. The olive oil is very green, and when I checked it at 12pm today it was a red wine color. Not brown at all.

#690 Allen Walters

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Posted 10 June 2012 - 11:18 PM

This is how it looks nowAttached File  IMG_20120610_190540.jpg   59.05KB   14 downloads

Edited by Allen Walters, 10 June 2012 - 11:19 PM.






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