3585 replies to this topic

[Anthony_Loera]

Hi,

The centrifuge is rated at 4420 g.... (at least the one i was considering)

Let me see if i can find a similar one that tops out at 5000 gs.

The filtering... now that is something i am still looking at. Any suggestions on that end would be appreciated!

Cheers
A

Circle me: https://profiles.goo...236572014252197

[niner]

I don't see how this is like cold fusion. It's consistent with lots of other fullerene work. We have reasonable hypotheses as to mechanism of action. After seeing reports of life extension in multiple species, including mammals given water soluble fullerols or acid derivatives, it doesn't surprise me that the fatty acid adducts are a lot better. I think that the odds of them extending human lifespan are reasonable. I think the odds that they will increase it by 90% are vanishingly small. Skepticism is appropriate, but not knee-jerk skepticism.

BTW, there is still research going on in the field now known as 'Low Energy Nuclear Reactions' or some such thing. A lot has been learned since 1989. Unfortunately, the field is very greatly hampered by public perception that "Cold Fusion is a Fraud". It was never a fraud. A debacle, yes, but not a fraud, nor a scandal.

[Junk Master]

Besides, even C60 just turns out to be a better version of PQQ that would be exciting.

[nowayout]

I would be the first to wish that this direction of research is successful.

One thing that bothers me, though, is that the median life span of Wistar rats is known to be around 30 months. That the control rats lived for 22 months, so much less than the known median lifespan, rings alarm bells despite the small size of the study. For a study to be convincing, the known median lifespan should at least be approximated in the control group, otherwise it raises questions about any number of possible problems, for example related to husbandry, lack of blinding causing differential treatment between groups, size of study, etc.

If the difference is solely due to a size effect, and we ascribe the fact that the control group lived 30% shorter than the expected 30 months to be a non-significant statistical variation, then the fact that the buckyball group lived 30% longer than the expected 30 months is likely also ascribable to a non-significant statistical variation. This would void the claimed confirmation of the hypothesis, because given the wide error bar, there is a 50/50 chance that the outcome could have gone the opposite way.

Edited by viveutvivas, 15 May 2012 - 09:58 PM.

[Turnbuckle]

I would be the first to wish that this direction of research is successful.

One thing that bothers me, though, is that the median life span of Wistar rats is known to be around 30 months. That the control rats lived for 22 months, so much less than the known median lifespan, rings alarm bells despite the small size of the study. For a study to be convincing, the known median lifespan should at least be approximated in the control group, otherwise it raises questions about any number of possible problems, for example related to husbandry, lack of blinding causing differential treatment between groups, size of study, etc.

If the difference is solely due to a size effect, and we ascribe the fact that the control group lived 30% shorter than the expected 30 months to be a non-significant statistical variation, then the fact that the buckyball group lived 30% longer than the expected 30 months is likely also ascribable to a non-significant statistical variation. This would void the claimed confirmation of the hypothesis, because given the wide error bar, there is a 50/50 chance that the outcome could have gone the opposite way.

A Japanese paper gives the average lifespans for male Wistar Mishima and male Wistar/Lobund rats as 731 days and 707 days respectively. That’s 24 months and 23 months, which isn’t far off from this set of male “Wistar” rats.

[HighDesertWizard]

re: the biological mechanism underlying this Frickin'-Fuller rat study... The discussion thus far is beyond my DiY expertise (inflammation, especially of the 5-Lipoxygenase--5-LO--variety). That said, I have a few thoughts that others, more knowledgeable, might find useful. Tell me what you think.

First, and to get your attention, I note the following...

• Just a few months ago, the LEF made the case that 5-LO was implicated in 7 of the 10 most lethal diseases in the US.

• Faster than normal aging--10 times faster?--takes place during extended Space Flights. A study conducted on the International Space Station, published just a few weeks ago, found that 5-LO was implicated as a potential key cause of accelerated aging. Summary here.

• Well, and there’s this... For me, by 2009, the science underlying the importance of 5-LO had gotten to be so voluminous that I could no longer stand it and prepared an FB Note about the diseases it’s implicated in in December 2010, before the LEF article appeared.

Ok, so what does this have to do with this Frickin'-Fuller rat study?

I don’t have familiarity with the mtDNA literature to evaluate the relevance of the following. I just think these thoughts might be important and so I provide them.

• Olive Oil (EPA/DHA) is a profound inhibitor of the 5-Lipoxygenase--Leukotriene B4 Inflammatory Cascade. The point is... The Olive Oil choice as a solvent is not inconsequential to the study result.

• The OLIVE OIL--C60 Study authors say, in the abstract itself, they believe the result has something to do with attenuating OxIdative Stress.

• Oxidative Stress implicates ROS (Reactive Oxygen Species) are at work and Mitochondrial DNA impact (about which I know nothing).

• But mtDNA impact implicates P53, the Guardian of the Genome, right?

So this study should be noted...

---------
5-Lipoxygenase regulates senescence-like growth arrest by promoting ROS-dependent p53 activation

5-Lipoxygenase (5LO) is involved in the production of leukotrienes and reactive oxygen species (ROS) from arachidonic acid. Its strong activation has been associated with several diseases like cancer and neurodegeneration. Here we show that 5LO activity increases during senescence-like growth arrest induced by oncogenic ras or culture history in both human and mouse embryo fibroblasts. Overexpression of 5LO promotes senescence-like growth arrest via a p53/p21-dependent pathway, and this occurs independently of telomerase activity. 5LO stabilizes p53 through phosphorylation at Ser15 and increases expression of the p53-transcriptional target p21. This is achieved by regulating ROS production. Indeed, ROS are increased in 5LO-arrested cells. Antioxidants and a low oxygen environment prevent 5LO-induced growth arrest. Finally, 5LO inhibition reduces the growth arrest induced by oncogenic ras or culture history and these effects are neutralized by the addition of exogenous ROS. These data link the 5LO pathway to oxidative crises of primary fibroblast and suggest that the ability of 5LO to induce senescence-like growth arrest may be important in the pathogenesis of 5LO-associated disorders.
---------

I presume to post these thoughts, not because I understand how they’re relevant but because, just this year...

• the LEF says 5-LO is important
• an aging in space study says 5-LO may be a major driver of aging processes
• a study of aging finds that rats, high on C60 Fullerenes dissolved in a 5-LO inhibitor, get their life spans doubled.

Tell me, if you can, that this is not yet another study about the profound health effects of managing 5-LO...

8-)

I didn't provide the link to the ISS Study Summary. Here it is... http://www.sciencedaily.com/releases/2012/04/120430105404.htm

Edited by wccaguy, 16 May 2012 - 12:07 AM.

[niner]

• the LEF says 5-LO is important
• an aging in space study says 5-LO may be a major driver of aging processes
• a study of aging finds that rats, high on C60 Fullerenes dissolved in a 5-LO inhibitor, get their life spans doubled.

I didn't get the impression from the space study that 5-LO was a major driver of aging processes. Maybe a minor one. There's at least one pretty decent 5-LO inhibitor already being used in humans. It's called Singulair, made be Merck. I took it for several years as part of my therapy for atypical asthma (allergy-driven inflammation without reactive airways). I didn't notice a thing from it, and eventually dropped it. It seems to me that if 5-LO inhibition was the magic bullet, I'd have seen something. My allergist claims that for some people, the effect of adding a 5-LO inhibitor is huge, but I wasn't one of them.

As for the relevance to the rat study, my question would be: Quantitatively, how potent a 5-LO inhibitor is olive oil? I don't think that's considered a major activity. That said, the vehicle controls lived a lot longer than the base controls. We know olive oil is good for us, but that result seemed too big. I consider that a problem with this small study, but maybe the rats really like being force-fed a bizarre variant of the Mediteranean diet.

[HighDesertWizard]

I didn't get the impression from the space study that 5-LO was a major driver of aging processes. Maybe a minor one. There's at least one pretty decent 5-LO inhibitor already being used in humans. It's called Singulair, made be Merck. I took it for several years as part of my therapy for atypical asthma (allergy-driven inflammation without reactive airways). I didn't notice a thing from it, and eventually dropped it. It seems to me that if 5-LO inhibition was the magic bullet, I'd have seen something. My allergist claims that for some people, the effect of adding a 5-LO inhibitor is huge, but I wasn't one of them.

As for the relevance to the rat study, my question would be: Quantitatively, how potent a 5-LO inhibitor is olive oil? I don't think that's considered a major activity. That said, the vehicle controls lived a lot longer than the base controls. We know olive oil is good for us, but that result seemed too big. I consider that a problem with this small study, but maybe the rats really like being force-fed a bizarre variant of the Mediteranean diet.

niner... Thanks for the reply. Some thoughts...

- Singulair is a prescribed drug. There are innumerable natural 5-LO inhibitors of various types. Many healthy substances we take for granted as healthy are especially healthy just because they inhibit 5-LO. (Omega 3s, Cocoa, Curcumin, Boswellia, Quercetin, even Resvertatrol, etc.) I don't see that the use of a 5-LO inhibition drug is necessary until the natural means for inhibiting 5-LO have been exhausted.

- About the importance of 5-LO... I'm just recently back to this forum. I'll create a thread about 5-LO soon. In the meantime, I encourage those not familiar with how important it is to check out the first few links in my post above. (The LEF says 5-LO is implicated in 7 of the 10 most lethal diseases in the US.)

- Olive Oil is a 5-LO inhibitor. I don't have time now to post studies showing this. I can easily do that at some point if it needs doing. I thought I read that a high quality Olive Oil was used in the Buckyball study. That could make a difference.

- But my main point had been to emphasize the choice of a high quality, known 5-LO inhibitor as the solvent for the Fullerene C60s. This would NOT be relevant or interesting to the mechanism question of a presumed actual life span increase in this study's rats if multiple studies hadn't already found a relationship between 5-LO inhibition and a reduction in ROS with implied impacts on Telomeres and Telomerase.

- But 5-LO actually has been found to inhibit ROS in multiple studies. It is a profound inhibitor of TNF-a. Study links below about ROS inhibition.

- So my question/point is this... Could it be that some unique feature of the combination of a 5-LO solvent with the Buckyballs is what extended life span? I believe I read upthread that the Buckyballs might clump in the mitochondria for some extended period. If that's true, then the combination with the Olive Oil would maintain 5-LO inhibition in the mitochondria for that same period, hence, perhaps clearing out the ROS.

I'm not certain this is what's going on that explains a presumed actual life span increase. I'm not familiar with the language and the meaning of much of the science around mtDNA and telomeres. I'm merely putting this point on the table for discussion: Let's not forget the importance of the Solvent. 8-)

Just a few of dozens of study links about 5-LO and ROS inhibition...

http://www.nature.co...s/7600502a.html
http://www.sciencedi...006295298003207
http://ajpgi.physiol...9/6/G1137.short
http://en.scientific...ns.org/57531507
http://informahealth...715761003758122

ROS and Telomere/Telomerase links...
http://www.sens.org/node/1538 -->> perhaps 5-LO can do to ROS what Telomerase can't
http://jcs.biologist...121/7/1046.full
http://www.ncbi.nlm....pubmed/14963003

[HighDesertWizard]

This figure helped me to better understand the question I've raised above. If I'm not mistaken, it underscores the importance of the point I'm trying to make.

Olive Oil is a 5-LO inhibitor. 5-LO inhibition reduces ROS, independent of P53. I cannot escape the conclusion that the choice of the Olive Oil solvent, per se, cannot be dismissed as being unimportant to the Buckyball study conclusion. At least not yet.

Thoughts?


The graphic figure is from this study...

http://www.ncbi.nlm....pubmed/20729567

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

Anthony Loera: Might this be THE hint about HOW to think about the choice of a Solvent for the Fullerene C60s? The solvent must be a 5-LO inhibitor. Perhaps an even more effective one than Olive Oil exists?

I dunno. I'm just asking questions.

8-)

Edited by wccaguy, 16 May 2012 - 12:41 PM.

[testerer]

Here's a public report from 2008 by the Russian team on solubility in natural oils:
http://www.spbcas.ru/Kozyrev/publications/reports/Fulleren's%20Interaction/Fulleren's%20Interaction.htm

[testerer]

Another interesting article, pointing that nano-C60 (nC60) which is C60+water (not oil) complex, is toxic (oxidative), though toxicity can be prevented by adding ascorbic acid or other antioxidants. Other studies also talk about several hydrophilic C60 complexes being oxidative in presence of optical stimuli, will try to find those.

http://nanonet.rice....ytotoxicity.pdf

[testerer]

Summary of studies: Caridovascular effects of water-soluble C60 derivatives:

http://www.kopfinstr...s/Carrier63.pdf

[testerer]

Another interesting study investigating water-soluble C60 derivative virucidal activity.
From results:

Although additional studies of the
mechanism of antiviral action of this compound are war-ranted, our results and those of Friedman et al. suggest that
the compound inhibits HIV-1 RT and DNA polymerase a
and selectively inhibits HIV-1 protease in cell-free systems
(Table 1) (6). This compound can be viewed as a lead in the
discovery of additional water-soluble fullerenes with greater
virucidal and antiprotease activities and should also stimu-late biochemical and pharmacological studies to determine
the metabolic disposition of this novel class of compounds in
living systems.

http://www.ncbi.nlm....cles/PMC188048/

[HappyPhysicist]

I am preparing a batch of C60/MCT oil with a concentration of 0.5 mg/mL for kicks. It has a much lower viscosity and should be easier to filter at least. The big question is whether there is something about olive oil that makes it so effective (assuming the rat study results are correct).

[testerer]

Here's a European company that could probably be contracted for some of the operations (centrifuge, filtration) of the C60-oil solution in europe.
http://particular.eu..._particular.pdf

[Turnbuckle]

Another interesting article, pointing that nano-C60 (nC60) which is C60+water (not oil) complex, is toxic (oxidative), though toxicity can be prevented by adding ascorbic acid or other antioxidants. Other studies also talk about several hydrophilic C60 complexes being oxidative in presence of optical stimuli, will try to find those.

http://nanonet.rice....ytotoxicity.pdf

Notice that they first dissolved the C60 in tetrahydrofuran and their test to determine that there was none left seems suspect. If C60 dragged any THF into the mitochondria, that would not be good.

[HighDesertWizard]

I am preparing a batch of C60/MCT oil with a concentration of 0.5 mg/mL for kicks. It has a much lower viscosity and should be easier to filter at least. The big question is whether there is something about olive oil that makes it so effective (assuming the rat study results are correct).

HappyPhysicist... I assume that by MCT Oil you mean Medium Chain Triglyceride Oil...

I take it from posts immediately prior to yours that the choice of solvent is not a trivial one.

To my way of thinking, here's what we know from the study we're discussing and from other sources...

• Olive Oil was used as a solvent
• Olive Oil is a 5-Lipoxygenase (5-LO) Inflammatory Pathway Inhibitor (google it to see for yourself)
• 5-LO inhibits ROS
• ROS inhibition has longevity implications (see the diagram I posted)
• To my mind, the links I've provided and the diagram I posted in my last thread provide a plausible *explanation* for the C60/Olive Oil study result
• The impact of 5-LO on ROS makes sense as a part of that *explanation*

I am NOT arguing that the explanation I've put on the table is the correct or only one. But I haven't seen a better one yet.

I wouldn't do this experiment on myself, but I understand the rationale related to your health that is driving your effort. I would do the same thing if I were in your position.

That said, I think it's a misstep to use anything other than a clear 5-LO inhibitor as a solvent. WHY NOT use precisely what the study used (Olive Oil) when there is a plausible argument, derived from studies, for the solvent they used? Or, at least, WHY NOT use a solvent for which 5-LO inhibition pops up immediately when doing a google search?

I just don't see 5-LO inhibition popping out for MCT Oil. Maybe I'm missing something... If I am, I stand corrected... But, in that case, prove it to me... I hope you see the value in getting this kind of feedback...

Just my two cents... I hope my posts and thoughts provide you with feedback that provides benefit.

All my best to you and yours.

8-)

Edited by wccaguy, 16 May 2012 - 03:28 PM.

[HappyPhysicist]

wccaguy,

These are all good points. Why would one deviate from the C60 preparation in the rate study? Well, if it wasn't so had to prepare I wouldn't. What I like about MCT is that it is transparent and after mixing with C60 has the signature purple color of C60 solutions. This gives me an added level of comfort that the solution is prepared properly and if I decide to skip the filter and centrifuge steps at least the color is a good indication that the vast majority of C60 is dissolved and not in the form of aggregates.

There are just too many unknowns here to make an educated decision. Its all guesswork at this stage.

Edited by HappyPhysicist, 16 May 2012 - 03:39 PM.

[HighDesertWizard]

There are just too many unknowns here to make an educated decision. Its all guesswork at this stage.

There are a lot of unknowns. I agree. There are also a lot things we do know. We know that the solvent choice is a non-trivial one...

You're taking some risk here in doing this n=1 experiment for understandable reasons. You're putting time and energy into doing this. IMHO, it's a mistake to take a short cut on the 5-LO oil question.

(My background for being able to say this: I've spent a couple thousand hours studying the 5-LO pathway. I have posted prolifically about this topic at Dr. Davis' TrackYourPlaque.com forum. I know more than a little about the importance of 5-LO. And (see the link in my first post) the LEF says its important too...)

Let's be clear... In using MCT Oil and given the evidence about the importance of 5-LO to human health and oxidation, you're NOT actually designing your experiment in a way that is consistent with the Buckyball study for a key study dimension, i.e., the solvent choice...

Tell me you're using an Omega 3 Oil... EPA/DHA... My response would be a positive one...

To be clear, my purpose here is not to give you a hard time for no reason but to provide you with informed feedback about a detail I have expertise about...

All the best...

[HappyPhysicist]

wccaguy,

The value of these forums and what make them in some ways superior to the traditional academic paradigm is the open real time debate, so I encourage this line of questioning. I only wish academics would engage is such open debate but I think their egos get bruised too easily.

You are definitely correct that I will likely learn little or noting by taking C60/MCT for a week, but I am going to give it a try anyway. Currently I am taking C3/OO (Olive Oil) and plan to do it primarily.

Thanks,

HP

[HighDesertWizard]

Ok. I hope you'll reconsider the decision to use MCT...

I believe WHAT you dissolve the Buckyballs in is more important than the HOW or the DEGREE of dissolving them... But I'm not going to preach about it any more.

Moving on.... Omega 3 Fatty Acids (EPA/DHA) inhibit 5-LO. Is it possible for you to shift your effort to Omega 3s? If you're just looking to vary the solvent, try some Omega 3 Oil you can purchase online...

[niner]

I am preparing a batch of C60/MCT oil with a concentration of 0.5 mg/mL for kicks. It has a much lower viscosity and should be easier to filter at least. The big question is whether there is something about olive oil that makes it so effective (assuming the rat study results are correct).

I think there is something special about olive oil; most of the fatty acids are unsaturated. This may be particularly important in the case of the doubly (or higher) unsaturated fatty acids, which may engage in a Diels-Alder coupling with the fullerene, with the fullerene acting as the dienophile. I read somewhere about a mechanism whereby a fullerene could link to an unsaturated [Edit: saturated(!)] oil (like MCT); it involved a peroxide intermediate. You might need to let the MCT/fullerene sit around for a couple weeks with some oxygen exposure to help that along. Keep us posted on how effective the MCT seems to be in terms of dissolution of the C60, and what colors you get.

My working assumption at present is that C60 reacts with the fatty acids in various oils, forming what I think is the active species, a fatty acid C60 adduct that can become an integral part of a biological membrane, including the mitochondrial membrane. I need to learn a lot more about the chemistry of this reaction, the structure and stability of the adducts, and their biological fate before I can say that this is more than an educated guess.

[Further edit: As long as I'm editing, I'll note that in the lipid chart I posted somewhere below, it lists coconut oil as 91% saturated and 9% unsaturated. Therefore, you might still form adducts without resorting to peroxide chemistry.]

Edited by niner, 17 May 2012 - 12:09 PM.

[niner]

I'm having a hard time with the 5-LO hypothesis. For one thing, olive oil isn't a very potent 5-LO inhibitor. The most potent component, hydroxytyrosol, has an EC50 of 15 uM. You would need to drink a LOT of olive oil to hit a concentration like that. OTOH, Singulair is a very potent 5-LO inhibitor, and while it is a good anti-inflammatory for some people, I've not seen any epidemiology showing anti-aging effects from it. We see life extension effects from water soluble fullerenes where there is nothing involved that could be called a 5-LO inhibitor.

[HappyPhysicist]

The experimental results show that the solubility of fullerenes appears to be dependent on the unsaturation level of the fatty acids composing the vegetable oils being lower in oils with higher unsaturation level. Franco Cataldo

Has anyone found a chart which orders the various oils in terms of "unsaturation" level?

It was already pointed out by someone in this forum that MCT is a pure saturated fat, and therefore has a low "unsaturation level" (If I understand that term correctly), and therefore should have a higher solubility of C60. The question is does the C60 still go along for the ride to the cells or mitochondria.

Edited by HappyPhysicist, 16 May 2012 - 06:34 PM.

[hav]

Here's a public report from 2008 by the Russian team on solubility in natural oils:
http://www.spbcas.ru...Interaction.htm

Looks like the link above got chopped. But I was able to locate it here. Seabuckthorn Oil sounds interesting... looks like it can dissolve 2x 53.75x as much C60 as Olive Oil and has some pretty interesting claimed qualities of its own with no reported undesirable effects I could find. But it is a bit more expensive.

Howard

Edited by hav, 16 May 2012 - 07:43 PM.

[Turnbuckle]

Here's a public report from 2008 by the Russian team on solubility in natural oils:
http://www.spbcas.ru...Interaction.htm

Looks like the link above got chopped. But I was able to locate it here. Seabuckthorn Oil sounds interesting... looks like it can dissolve 2x as much C60 as Olive Oil and has some pretty interesting claimed qualities of its own with no reported undesirable effects I could find. But it is a bit more expensive.

Howard

It's not at all clear what reacting C60 with a heated solvent would do, but it would surely act differently in vivo.

Edited by Turnbuckle, 16 May 2012 - 07:08 PM.

[hav]

Here's a public report from 2008 by the Russian team on solubility in natural oils:
http://www.spbcas.ru...Interaction.htm

Looks like the link above got chopped. But I was able to locate it here. Seabuckthorn Oil sounds interesting... looks like it can dissolve 2x as much C60 as Olive Oil and has some pretty interesting claimed qualities of its own with no reported undesirable effects I could find. But it is a bit more expensive.

Howard

It's not at all clear what reacting C60 with a heated solvent would do, but it would surely act differently in vivo.

I think I confused myself. If 43 mg of C60 dissolve into 1 ml of Seabuckthorn Oil that would be over 50x the ratio of .8 grams of C60 dissolving into a liter of Olive Oil. Cutting the oil volume needed by a factor of 50 would make dosage quite practical. Too bad there's no research on C60 with Seabuckthorn Oil.

Howard

[HappyPhysicist]

Something is not right. Cataldo's paper states a solubility of Castor oil, for instance, as 392 mg/l = 0.392 mg/mL, which is different from the Russian paper by about a factor of 100.

Edited by HappyPhysicist, 16 May 2012 - 07:49 PM.

[Turnbuckle]

Something is not right. Cataldo's paper states a solubility of Castor oil, for instance, as 392 mg/l = 0.392 mg/mL, which is different from the Russian paper by about a factor of 100.

They're heating it, and when you heat it you get a reaction. Then it's not a true solution anymore.

[hav]

The translation is so cryptic, I just couldn't get that part...

The fullerene's was entered into a liquid sample which then placed in the dispenser with composite ultrasonic and electromagnetic influence , temperature in which one could be changed.

Thanks for decoding.

Howard