635 replies to this topic

[ImmInst]

NOTE: this thread is about the paper by Baati et al, as and about the potential mechanisms of action. Many discussions are focussed on trying C^0 as a supplement at home, these can be found here and elsewhere in this subforum.
UPDATE 18th June 2012: This thread is for general scientific discussions only. Further anecdotal reports, banter etc. will be DELETED without warning


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A research group is claiming that fullerenes (C60, ingested and injected) greatly extend life span in mice; this is meeting with some considerable skepticism, given the degree of life extension and the lack of a plausible mechanism. "In the current study researchers fed the molecule dissolved in olive oil to rats and compared outcomes to a control group of rats who got plain olive oil. The main question they wanted to answer was whether chronic C60 administration had any toxicity, what they discovered actually surprised them. ... Here we show that oral administration of C60 dissolved in olive oil (0.8 mg/ml) at reiterated doses (1.7 mg/kg of body weight) to rats not only does not entail chronic toxicity, but it almost doubles their lifespan. ... The estimated median lifespan (EML) for the C60-treated rats was 42 months while the EMLs for control rats and olive oil-treated rats were 22 and 26 months, respectively. Using a toxicity model the researchers demonstrated that the effect on lifespan seems to be mediated by 'attenuation of age-associated increases in oxidative stress'." So what might be going on here? The average life span of the Wistar rats used is 2-3 years (24 - 36 months). This was a small study size, but that's no so important in determining whether you have an actual means of life extension if you can show that any of your study group lived much longer than usual - but it is important when it comes to the degree of life extension. If the study group is small, as it is here, using only a handful of rats, then the size of the effect can be much more readily distorted by chance. This line in the paper jumped out at me: "Before C60 administration, the rats were fasted overnight but with access to water." If they failed to fast the control group, then we're looking at yet another study that failed to control for calorie restriction, and this is actually largely an intermittent fasting study - which has certainly been shown to extend life in rats.

Link: http://extremelongevity.net/2012/04/16/chronic-buckyball-administration-doubles-rat-lifespan/

View the full article at Fight Aging
download the paper here

Edited by caliban, 19 April 2015 - 09:48 PM.

[8bitmore]

I agree that the study group is smallish but there's several interesting bits to be found in the full text (http://extremelongev...0-Fullerene.pdf) that are suggestive of interesting (possibly novel?) life extension properties of C60.

Here's what stood out for me when I read the study, let me know if I'm blatantly off on any of the points,

• It is interesting that the C60 is almost equally well absorbed orally as compared to IP injection (body cavity injection)
• C60 crosses the BBB - "the presence of significant amounts in the brain 24 hours after both oral and i.p. administrations under our experimental conditions (Table 2) confirms that solubilized C60 can cross the bloodebrain barrier"
• It might well be that compounds formed when C60 breaks down are instrumental to the health benefits, it is as of yet unknown how the animals process/eliminate the C60 - "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."
• The C60 seems to work by down-regulating the amount of anti-oxidant activity in the rats (both Catalase and SOD); especially when under stress from CCl4 (Carbon tetrachloride?). Yet, it still obviously manages to protect them very well.

Edited by 8bitmore, 17 April 2012 - 01:59 PM.

[Ampa-omega]

Buckyballs are just carbon but this is surprising... hell, in 20 years maybe diamonds will be the next selegiline.

wow i find this fascinating
neurogenesis from carbon buckyballs/nanotubes? how does that work?

Edited by Ampa-omega, 17 April 2012 - 05:37 PM.

[smithx]

http://www.sciencedi...142961212003237

Countless studies showed that [60]fullerene (C₆₀) and derivatives could have many potential biomedical applications. However, while several independent research groups showed that C₆₀ has no acute or sub-acute toxicity in various experimental models, more than 25 years after its discovery the in vivo fate and the chronic effects of this fullerene remain unknown. If the potential of C₆₀ and derivatives in the biomedical field have to be fulfilled these issues must be addressed. Here we show that oral administration of C₆₀ dissolved in olive oil (0.8 mg/ml) at reiterated doses (1.7 mg/kg of body weight) to rats not only does not entail chronic toxicity but it almost doubles their lifespan. The effects of C₆₀-olive oil solutions in an experimental model of CCl₄ intoxication in rat strongly suggest that the effect on lifespan is mainly due to the attenuation of age-associated increases in oxidative stress. Pharmacokinetic studies show that dissolved C₆₀ is absorbed by the gastro-intestinal tract and eliminated in a few tens of hours. These results of importance in the fields of medicine and toxicology should open the way for the many possible -and waited for- biomedical applications of C₆₀ including cancer therapy, neurodegenerative disorders, and ageing.

[tunt01]

haven't gone through the whole thing, but on the surface it seems less than enthralling. sample size of only 6 rats per group. and the data seems more compelling for olive oil than fullerene, per se. they really should have had a 4th group for water+fullerene.

[MrHappy]

What changes in lifespan and neurite outgrowth did they get with just the olive oil?

[8bitmore]

[...] and the data seems more compelling for olive oil than fullerene, per se. [...]

On the surface yes, but when you look at the data on page 8 (fig.5 & 6) it becomes clear that olive oil unlike the C60 does nothing to protect the rats against CCl4 intoxication; i.e. it does not modulate the antioxidant response at all, not with Glutathione, SOD or Catalase. Also, obviously it only modulates the lifespan marginally (18%) whereas the C60 treated rats are living 90% longer. I think these data definitely warrants a followup study; the authors mention that C60 is produced by the ton now (what for I wonder, probably some industrial purpose?) so it should be relatively straightforward to try and replicate their results with a larger group of animals.

Also, on a personal note: I really find the images on page 7 rather incredible, those C60 treated rat livers look SO healthy compared to the oil/control groups.

[8bitmore]

What changes in lifespan and neurite outgrowth did they get with just the olive oil?

I just answered that (well, not the neurite outgrowth question, but that's because they do not deal with that directly in the research paper*) in Bioscience forum -->
http://www.longecity...675#entry511675

*they did however reference a paper that does:

Syntheses of water-soluble [60]fullerene derivatives and their enhancing effect on neurite outgrowth in NGF-treated PC12 cells.
Tsumoto H, Kawahara S, Fujisawa Y, Suzuki T, Nakagawa H, Kohda K, Miyata N
Bioorg Med Chem Lett. 2010 Mar 15; 20(6): 1948-52

Water-soluble [60]fullerene (C(60)) derivatives were synthesized to examine their bioactivities. PC12 cells were used as a model of nerve cells and the bioactivities of synthesized C(60) derivatives together with some reported ones were tested. Among the compounds tested, C(60)/(gamma-CyD)(2), C(60)-bis(gamma-CyD) (5) containing C(60)-mono(gamma-CyD) (5'), and C(60)/PVP were sufficiently soluble in water and showed an enhancing effect on the neurite outgrowth of NGF-treated PC12 cells.

[niner]

This is an amazing result, at least it will be once it's replicated with a larger sample size and attention to diet in order to exclude inadvertent CR effects. C60 fullerenes come in a variety of purities; we would want to know what else was in it and if it was toxic. At this site, (Fullerenes for Less!) the following prices are given:

Amount >99.0% >99.5% >99.98% Sublimed
50-99g $24/g $26/g $58/g $110/g

I'd hope that we wouldn't need the sublimed stuff. 50g of 99.5% is $1300. That's $2.60/day for a 100 mg dose, which would be a little less than the dose used in the paper for most people. If you apply an interspecies scaling factor of 6, then it's 43 cents a day. Not too bad. If the interspecies scaling is legitimate for a therapeutic dose, as opposed to the toxicity tests for which such scaling was developed, then even the sublimed purity would cost less than a lot of people spend on their daily coffee.

My favorite thing about this paper? They used the phrase "paramount potentialities" in the first sentence.

[stephen_b]

Dose might play a role too. What are the chances they hit upon the optimal dose for this study?

[8bitmore]

[...]
Amount >99.0% >99.5% >99.98% Sublimed
50-99g $24/g $26/g $58/g $110/g

I'd hope that we wouldn't need the sublimed stuff.
[...]

Do you know what the "sublimed" denominator stands for? (beyond the layman's i-can-infer-its-been-distilled-a-lot meaning of it). It would be interesting to know what the contaminants are, novel nano-diseases can be a bit boring..!

Also, has anyone found-out/seen what the industrial use is for this stuff?

[Invariant]

Another post on extremelongevity on this study: http://extremelongev...span-extension/
The authors claim there probably wasn't any CR going on.. This could be big, but we definitely need human testing. Such evidence might be forthcoming or already in existence, since authorities seem to be quite concerned with potential hazards of nanomaterials (and rightly so I guess).

[Junk Master]

Has anything else almost doubled lifespan in mice?

[Junk Master]

I also wonder, if we are assuming the result is due to antioxidant properties, if the results would be even more significant with cyclical dosing.

[smithx]

Or maybe it's all a mistake and a really bad idea:

Eva Oberdorster of Southern Methodist University in Dallas, US, who led the study, found modest concentrations of buckyballs in water caused significant harm to two aquatic animals. Water fleas were killed by the addition of the tiny carbon balls, and fish showed up to a 17-fold increase in brain damage compared with unexposed animals.

http://www.newscient...ge-in-fish.html

[8bitmore]

Or maybe it's all a mistake and a really bad idea:

Eva Oberdorster of Southern Methodist University in Dallas, US, who led the study, found modest concentrations of buckyballs in water caused significant harm to two aquatic animals. Water fleas were killed by the addition of the tiny carbon balls, and fish showed up to a 17-fold increase in brain damage compared with unexposed animals.

http://www.newscient...ge-in-fish.html

Thank you for finding that study; it extremely important to temper these sort of threads so we don't inadvertently end up enticing some idiot to eat up a dosage of "chemical-x" based on some very vague assumption that it could make them live 90% longer (it could potentially, but then it could also potentially give them brain damage instead).

About the actual study mentioned in the New Scientist article you linked; here is the abstract of some of the research (see end of post), what I find interesting about it is that they used nC60 and not C60 which can fully explain the toxic effects (and coincidentally also demonstrates that it is probably absolutely crucial to suspend these molecules in olive oil before using them so they stay C60 instead of nC60). The following paragraph from this study (http://cohesion.rice...plibrary/96.pdf) highlights the point nicely:

"Initially, the hydrophobicity of C60 (water solubility <10-9mg/L) was thought to limit its interactions with biological systems and oral administration, skin application, or injection of C60 in rats and other cellsystems revealed no acute toxicity. However, once C60 is introduced into water (either via solvents or by extensive stirring) it forms stable nanoscale suspended aggregates known as fullerene water suspensions (FWS) or nC60 and becomes biologically active. nC60 is highly toxic to eukaryotic cell lines, Daphnia magna and fish."

In other words, any animal/human who is/were to ingest C60 better well make sure that it is suspended in oil like the authors of the life extending 2012 study did (study linked again for good measure: http://extremelongev...0-Fullerene.pdf) and abstain from drinking water close to the ingestion of the compound.. that being said humans are roughly 60-70% water.. it is a tricky but potentially extremely positive situation in relation to life extension.

Manufactured nanomaterials (fullerenes, C60) induce oxidative stress in the brain of juvenile largemouth bass.
Oberdorster E
Environ Health Perspect. 2004 Jul ; 112(10): 1058-62

Although nanotechnology has vast potential in uses such as fuel cells, microreactors, drug delivery devices, and personal care products, it is prudent to determine possible toxicity of nanotechnology-derived products before widespread use. It is likely that nanomaterials can affect wildlife if they are accidentally released into the environment. The fullerenes are one type of manufactured nanoparticle that is being produced by tons each year, and initially uncoated fullerenes can be modified with biocompatible coatings. Fullerenes are lipophilic and localize into lipid-rich regions such as cell membranes in vitro, and they are redox active. Other nano-sized particles and soluble metals have been shown to selectively translocate into the brain via the olfactory bulb in mammals and fish. Fullerenes (C60) can form aqueous suspended colloids (nC60); the question arises of whether a redox-active, lipophilic molecule could cause oxidative damage in an aquatic species. The goal of this study was to investigate oxyradical-induced lipid and protein damage, as well as impacts on total glutathione (GSH) levels, in largemouth bass exposed to nC60. Significant lipid peroxidation was found in brains of largemouth bass after 48 hr of exposure to 0.5 ppm uncoated nC60. GSH was also marginally depleted in gills of fish, and nC60 increased water clarity, possibly due to bactericidal activity. This is the first study showing that uncoated fullerenes can cause oxidative damage and depletion of GSH in vivo in an aquatic species. Further research needs to be done to evaluate the potential toxicity of manufactured nanomaterials, especially with respect to translocation into the brain.

Edited by 8bitmore, 19 April 2012 - 08:40 AM.

[smithx]

Very interesting data.

Even if C60 suspensions are ok initially do we have any evidence they're fine for 40 or 50 years or more?

The animals in the study live less than 4 years, so if the C60 breaks down into something toxic or clumps to form nC60 after 6 or 10 or 20 years, we wouldn't see it in such studies.

I'd be concerned that some percentage of the C60 would sit around, in a human brain, for example, until it either aggregates and becomes toxic, or breaks down into something else which is toxic.

[8bitmore]

Very interesting data.

Even if C60 suspensions are ok initially do we have any evidence they're fine for 40 or 50 years or more?

The animals in the study live less than 4 years, so if the C60 breaks down into something toxic or clumps to form nC60 after 6 or 10 or 20 years, we wouldn't see it in such studies.

I'd be concerned that some percentage of the C60 would sit around, in a human brain, for example, until it either aggregates and becomes toxic, or breaks down into something else which is toxic.

Hi smithx, in regards to the long term impact of C60; no we do obviously not have evidence but then again just simply reading the abstract from the Prolongation Study provides us with the following observation: "Pharmacokinetic studies show that dissolved C60 is absorbed by the gastro-intestinal tract and eliminated in a few tens of hours". And again: "Previous results obtained after i.p. administration of large doses
of micronized C60[21] or intratracheally instilled C60 suspensions [43] showed that the clearance of C60from organs can take several days.". This all boils down to: the C60 is eliminated alright: if there has been gradual accumulation of the molecules then it would have shown in the subsequent dissection of the rats. This will probably be even more clearly illuminated in a yet-to-be-published-study since the authors state on page 9 that: "A complete biodistribution study including intestine, skin, bone and fatty tissue is in progress in our laboratory."

[stephen_b]

because i thought it was totally misleading that they claim a doubling of lifespan where olive oil alone produced ~70-80% of the same benefits. i must be missing something.

The article has this quote from the paper:

“The estimated median lifespan (EML) for the C60-treated rats was 42 months while the EMLs for control rats and olive oil-treated rats were 22 and 26 months, respectively,” they write.

26 months is 62% of 42 months.

[tunt01]

26 months is 62% of 42 months.

it's statistical sleight of hand. he's taking the median number when obviously the dispersion in the olive oil rats wasn't nearly as uniform as the other groups. i also think with an n=6, it makes me really think very little of these results other than semi-interesting.

but who knows... haven't looked closely enough. will this weekend. thx.

[PWAIN]

I thought 42 weeks was around normal maximal lifespan for rats? What was the maximum lifespan of these rats?

[hav]

About the actual study mentioned in the New Scientist article you linked; here is the abstract of some of the research (see end of post), what I find interesting about it is that they used nC60 and not C60 which can fully explain the toxic effects (and coincidentally also demonstrates that it is probably absolutely crucial to suspend these molecules in olive oil before using them so they stay C60 instead of nC60). The following paragraph from this study (http://cohesion.rice...plibrary/96.pdf) highlights the point nicely:

"Initially, the hydrophobicity of C60 (water solubility <10-9mg/L) was thought to limit its interactions with biological systems and oral administration, skin application, or injection of C60 in rats and other cellsystems revealed no acute toxicity. However, once C60 is introduced into water (either via solvents or by extensive stirring) it forms stable nanoscale suspended aggregates known as fullerene water suspensions (FWS) or nC60 and becomes biologically active. nC60 is highly toxic to eukaryotic cell lines, Daphnia magna and fish."

In other words, any animal/human who is/were to ingest C60 better well make sure that it is suspended in oil like the authors of the life extending 2012 study did (study linked again for good measure: http://extremelongev...0-Fullerene.pdf) ...

Looks like they went to allot of trouble to make sure that all the C60 was dissolved in the olive oil and that none was in suspension:

2.1. C60-olive oil solution preparation

Virgin olive oil is obtained from a Chemlali Boughrara cultivar from Tunisia
planted in the Sahel area. C60 (purity 99.98%) was obtained from SES Research
Corporation (USA) and used without further purification.
Fifty mg of C60 were dissolved in 10 ml of olive oil by stirring for 2 weeks at
ambient temperature in the dark. The resulting mixture was centrifuged at 5.000 g for
1 h and the supernatant was filtered through aMillipore filter with 0.25 mmporosity.

I wonder if the toxicity results might be different without the filter and centrifuge steps.

Howard

[hav]

This other study (cited as reference #52) indicates that C60 in suspension in saline or corn oil does have increased toxicity:

http://www.ncbi.nlm....les/PMC2685830/

Also, the C60 olive oil study mentions on the last page:

It is to be stressed that dissolved C60 appears hundred times
more active than when it is in suspension [21]. In fact the action of
soluble C60 is immediate while that of suspended C60 is delayed
because it has to be dissolved to act.

So I guess making sure its all dissolved is kind of important.

Howard

[malbecman]

Just digging around on the web about this brought up this interesting find about the authors of the original paper using the same histology image twice in the paper. The
blog writer has requested clarification from the journal. The comments at the end of the blog are interesting as well.

http://pipeline.cora...s_a_problem.php

[LeonardElijah]

i'd sooner shoot up AICAR than put buckyballs into my body. without knowing the mechanism of action, it's just crazy to me. i gotta look at this study more closely this weekend, because i thought it was totally misleading that they claim a doubling of lifespan where olive oil alone produced ~70-80% of the same benefits. i must be missing something.

Not knowing the mechanism of action makes it exciting.

Suppose it simply makes sense that taking NGF extends neurites and taking Ashwaghanda extends neurites, but an otherwise inert substance affects the neural environment in some manner that produces a similar effect. What effect does the buckyball have on the environment? Understanding this effect, what are the implications for the apothecary's cupboard?

Edited by LeonardElijah, 20 April 2012 - 06:17 PM.

[Elus]

I hope this doesn't turn out to be a hoax or bad science (Thanks for pointing out that suspicious aspect of the paper, Malbecman). Bad science could undermine the credibility of the longevity movement.

Edited by Elus, 20 April 2012 - 07:36 PM.

[tunt01]

Just digging around on the web about this brought up this interesting find about the authors of the original paper using the same histology image twice in the paper. The
blog writer has requested clarification from the journal. The comments at the end of the blog are interesting as well.

http://pipeline.cora...s_a_problem.php

This doesn't surprise me at all. It looked to me like they spruced up these results.

If you look at comment #6 in the link you cited, he points out the longevity estimate issue that I mentioned earlier in the thread. They are using some statistical sleight of hand or just plain misrepresenting the data to get to a "doubling in lifespan".

Edited by prophets, 21 April 2012 - 03:33 AM.

[8bitmore]

Looks like they went to allot of trouble to make sure that all the C60 was dissolved in the olive oil and that none was in suspension:

2.1. C60-olive oil solution preparation

Virgin olive oil is obtained from a Chemlali Boughrara cultivar from Tunisia
planted in the Sahel area. C60 (purity 99.98%) was obtained from SES Research
Corporation (USA) and used without further purification.
Fifty mg of C60 were dissolved in 10 ml of olive oil by stirring for 2 weeks at
ambient temperature in the dark. The resulting mixture was centrifuged at 5.000 g for
1 h and the supernatant was filtered through aMillipore filter with 0.25 mmporosity.

I wonder if the toxicity results might be different without the filter and centrifuge steps.

Thanks so much for catching the distinction between "suspension" and "dissolvement" - for me they were one and the same when I wrote the post which is obviously NOT the case seeing the procedure required to make the C60 dissolve! Of course this changes the biological interactions radically: even just simple day-to-day things like Creatine Monohydrate intake for me is massively influenced by whether the particles are suspended or dissolved*

*dissolving Creatine Monohydrate completely in water can be achieved by making sure the water is roughly 45-50c anything less than that does not do the job.

Edited by Michael, 15 June 2012 - 04:33 PM.

[Brett Black]

A prior study also showing lifespan extension with fullerene, but less dramatic; this time in mice starting at 12 months of age and with much larger control and treatment groups:


1. Neurobiol Aging. 2008 Jan;29(1):117-28. Epub 2006 Oct 31.

A carboxyfullerene SOD mimetic improves cognition and extends the lifespan of
mice.

Quick KL, Ali SS, Arch R, Xiong C, Wozniak D, Dugan LL.

Department of Neurology, Washington University School of Medicine, St. Louis, MO
63110, United States.

In lower organisms, such as Caenorhabditis elegans and Drosophila, many genes
identified as key regulators of aging are involved in either detoxification of
reactive oxygen species or the cellular response to oxidatively-damaged
macromolecules. Transgenic mice have been generated to study these genes in
mammalian aging, but have not in general exhibited the expected lifespan
extension or beneficial behavioral effects, possibly reflecting compensatory
changes during development. We administered a small-molecule synthetic enzyme
superoxide dismutase (SOD) mimetic to wild-type (i.e. non-transgenic,
non-senescence accelerated) mice starting at middle age. Chronic treatment not
only reduced age-associated oxidative stress and mitochondrial radical
production, but significantly extended lifespan. Treated mice also exhibited
improved performance on the Morris water maze learning and memory task. This is
to our knowledge the first demonstration that an administered antioxidant with
mitochondrial activity and nervous system penetration not only increases
lifespan, but rescues age-related cognitive impairment in mammals. SOD mimetics
with such characteristics may provide unique complements to genetic strategies to
study the contribution of oxidative processes to nervous system aging.

PMID: 17079053 [PubMed - indexed for MEDLINE]

http://www.ncbi.nlm....pubmed/17079053
http://thebronxproje... Aging 2006.pdf (full text)

Edit: fixed link to full text

Edited by niner, 21 April 2012 - 09:22 PM.

[Brett Black]

A prior study also showing lifespan extension with fullerene, but less dramatic; this time in mice starting at 12 months of age and with much larger control and treatment groups:



1. Neurobiol Aging. 2008 Jan;29(1):117-28. Epub 2006 Oct 31.

A carboxyfullerene SOD mimetic improves cognition and extends the lifespan of
mice.

Quick KL, Ali SS, Arch R, Xiong C, Wozniak D, Dugan LL.

Department of Neurology, Washington University School of Medicine, St. Louis, MO
63110, United States.

In lower organisms, such as Caenorhabditis elegans and Drosophila, many genes
identified as key regulators of aging are involved in either detoxification of
reactive oxygen species or the cellular response to oxidatively-damaged
macromolecules. Transgenic mice have been generated to study these genes in
mammalian aging, but have not in general exhibited the expected lifespan
extension or beneficial behavioral effects, possibly reflecting compensatory
changes during development. We administered a small-molecule synthetic enzyme
superoxide dismutase (SOD) mimetic to wild-type (i.e. non-transgenic,
non-senescence accelerated) mice starting at middle age. Chronic treatment not
only reduced age-associated oxidative stress and mitochondrial radical
production, but significantly extended lifespan. Treated mice also exhibited
improved performance on the Morris water maze learning and memory task. This is
to our knowledge the first demonstration that an administered antioxidant with
mitochondrial activity and nervous system penetration not only increases
lifespan, but rescues age-related cognitive impairment in mammals. SOD mimetics
with such characteristics may provide unique complements to genetic strategies to
study the contribution of oxidative processes to nervous system aging.

PMID: 17079053 [PubMed - indexed for MEDLINE]

http://www.ncbi.nlm....pubmed/17079053
http://thebronxproject.com/Articles/7.)%20%20Neurobiology%20of%20Aging%202006.pdf