67 replies to this topic

[ClarkSims]

I just read skimmed this article several times,

http://www.fasebj.or.../23/7/2065.long

Senile hair graying: H2O2-mediated oxidative stress affects human hair color by blunting methionine sulfoxide repair

They say:

Accordingly, Met-S=O formation of Met residues, including Met 374 in the active site of tyrosinase, the key enzyme in melanogenesis, limits enzyme functionality, as evidenced by FT-Raman spectroscopy, computer simulation, and enzyme kinetics, which leads to gradual loss of hair color. Notably, under in vitro conditions, Met oxidation can be prevented by L-methionine.

There is a big problem with this analysis. They are using Methionine to prevent the oxidation of methione in an oxidizing environment. This will kind of work, if you have more methionine than H2O2, and if you are looking at the ratio of methionine to oxidized methionine. But if you are looking at the absolute value of the oxidized metionine, it is a monotonically increasing function the the amount of methionine and the amount of H2O2.

Morever metionine has no role in the functioning of Tyrosinase:


http://en.wikipedia....wiki/Tyrosinase

Tyrosinase catalyzes the change of Catechol into Quinone.
Oxidized Methionine (methionine sulfoxide) stops the enzyme from working.

To my ear, the solution is to reduce the amount of methionine, which will reduce the amount of methionine sulfoxide, increase the amount of other anti oxidants, like C60.

The only role I see for Methionine, is that it gets turned into Cystiene, which is a major component of hair:
http://www.ajichem.c...=397&h=219&as=1

So one should supplement Cystiene, and restrict Methionine.

In particular, I think a solution might be to mix C60 olive oil, with a small amount of DMSO, and use it as a hair tonic. The DMSO, should carry the C60-OO adduct into the skin, and the C60 should prevent the formation of H2O2, as well as neutralize any H2O2 which is there.

[Turnbuckle]

In particular, I think a solution might be to mix C60 olive oil, with a small amount of DMSO, and use it as a hair tonic. The DMSO, should carry the C60-OO adduct into the skin, and the C60 should prevent the formation of H2O2, as well as neutralize any H2O2 which is there.

Have you tried this?

[ClarkSims]

I haven't tried adding the DMSO yet.
I have been putting oo-c60 on my scalp and skin for about a month.
I have been on a methionine restricted diet for about 2 months.
I have been taking 2g of cystiene every day for about 2 months.

I just started looking for hairs changing colors with a magnifying glass. I will report any changes if I find them.

I have been wanting to add some dmso to the oo-c60 combination, but haven't had time.

Edited by ClarkSims, 03 January 2013 - 11:43 PM.

[ClarkSims]

Could someone tell me what Met 374 is?


Accordingly, Met-S=O formation of Met residues, including Met 374 in the active site of tyrosinase, the key enzyme in melanogenesis, limits enzyme functionality, as evidenced by FT-Raman spectroscopy, computer simulation, and enzyme kinetics, which leads to gradual loss of hair color.

[Turnbuckle]

I tried C60 in DMSO some time ago, and there was no solubility.

[zorba990]

You could try limonene.

[malbecman]

Met 374 refers to the 374th amino acid in the enzyme tyrosinase being a methionine (Met). Every amino acid has a 3 letter abbreviation. It's a shorthand way of labeling amino acids and their location in a protein's sequence.

Could someone tell me what Met 374 is?


Accordingly, Met-S=O formation of Met residues, including Met 374 in the active site of tyrosinase, the key enzyme in melanogenesis, limits enzyme functionality, as evidenced by FT-Raman spectroscopy, computer simulation, and enzyme kinetics, which leads to gradual loss of hair color.

[Turnbuckle]

You could try limonene.

That's a good idea. The solubility of C60 in limonene is quite good--about the same as in olive oil--and limonene itself has been touted as a hair growth stimulator.

[ClarkSims]

That is interesting. What would be a good ratio of C60-oo to limonene? For the DMSO I was thinking about 90% oo-c60 and 10% DMSO.

[ClarkSims]

Limonene sounds promising:

http://www.sciencedi...378517305008690

[YOLF]

This would make a great experiment and quite possibly attract alot of attention and research to the C60 molecule. The experiment definitely needs to be done right. Do you have a camera with a macro lens for taking detail pics up close? It would be nice to have a photographic record. I suggest taking the picture in the same place and under the same light, keeping around a spare set of CFLs that you only plug in to take the pics (the light output decreases significantly in even a months time and may be accompanied by a slight color temp shift). When you take the picture it should be in either a neutral pose or in a pose that you will easily be able to remember, like a pose from the thriller dance to ensure reproduceable light conditions in your photos. Check with friends. Macro lenses are common on some mid grade sony digitals and then there are the DSLRs that will have them. Sony, to my eyes always made the most functional macro lenses and they were on $200-300 cams. Try taking some close up pics of book text with autofocus, the more stuff in crystal clear focus, the better the macro will be.

I haven't tried adding the DMSO yet.
I have been putting oo-c60 on my scalp and skin for about a month.
I have been on a methionine restricted diet for about 2 months.
I have been taking 2g of cystiene every day for about 2 months.

I just started looking for hairs changing colors with a magnifying glass. I will report any changes if I find them.

I have been wanting to add some dmso to the oo-c60 combination, but haven't had time.

[niner]

One of the problems with trying to deliver c60-oo transdermally is that c60-oo is most likely a prodrug rather than the active agent. Olive oil is a glycerol triester, so when c60 reacts with one of the fatty acid chains, there's still a bunch of other junk attached to it, and it's a complex neutral molecule that isn't going to work well in a membrane. It's not until it encounters a lipase, which enzymatically cleaves the fatty acid from the glycerol backbone, that you end up with a free fatty acid with a c60 attached. The body normally encounters glycerol triesters in the diet, so you would expect to find lipases in the GI tract, but are they found in skin? At sufficient activity? I don't know. A possible way around this, if someone wants to experiment, would be to buy some ethyl oleate. This is the ester formed by condensing oleic acid with ethyl alcohol. You could probably react C60 with it the same way we do with olive oil. Instead of a triglyceride product, you'd get a mono-ester, which should be more soluble in DMSO, and should penetrate skin better. Instead of requiring a lipase, most any garden variety cytosolic esterase should be able to cleave it, so you should end up with the oleic acid adduct, once it got into the cell. The ethyl ester might even be more cell-permeable than the acid.

[zorba990]

I was thinking just C60 dissolved in limonene no olive oil.

[Turnbuckle]

I was thinking just C60 dissolved in limonene no olive oil.

You may find the straight stuff to be too aggressive. A while back, limonene dissolved the plastic test tubes I was using and poured out.

[niner]

I was thinking just C60 dissolved in limonene no olive oil.

Well, you could try it, bearing Turnbuckle's warning in mind. C60 might actually react with limonene, rather than just dissolve, since limonene has two double bonds. The end result would be pure hydrocarbon, rather than having a polar head group like a fatty acid adduct, so it wouldn't be locked into the membrane structure as well, but it would probably still work, at least at some level.

[NanoDoom]

Hello,

The other day, I took 25ml EVOO and 25ml Jacob Lab 90% DMSO (10% water) and mixed it in a glass container.
I put it in a dark cupboard at room temperature for around 24 hours, where it was stirred about five times in total.

The solution remained separated in DMSO/water and Oilve Oil parts.

I threw it out. It might take a lot longer and take a lot more stirring for it to mix. I don't know.


Have you considered replacing DMSO with Emu Oil?

For the past three weeks, I've been applying a mixture of 66% C60-OO and 33% Emu Oil to my receding hairline, where I also have some gray hair.
I use a 0.3mm Dermaroller/Microneedle roller for better skin penetration.

So far, no changes at all.

I'm also taking 1.5mg C60-OO orally, daily.


Emu Oil is said (by those who sell it...) to have the same properties as DMSO when it comes to pulling in other compounds into the skin. I have no idea whether this is true or not.

What I do know is that the Emu Oil seemed to mix with the C60-OO readily.
I don't know how/if adding Emu Oil to the mix will change the C60 itself. I'm hoping it will not.

[ClarkSims]

Niner raises a good point about the olive oil being a triglyceride of oleic acid.

Perhaps it would be best do dissolve the C60 into oleic acid, and then combine the oleic acid with something like limonene or DMSO, Emu Oil, or ethyl oleate for transdermal delivery.


http://www.dkhardwar...id-1-quart.html

I just dug up this paper

http://www.jlr.org/c...t/51/1/120.full

methyl-β-cyclodextrin might be a good thing to include in the mix also, to increase the rate at which the oleac acid - c60 adduct is incorporated into cell membranes.

Edited by ClarkSims, 04 January 2013 - 08:46 PM.

[ClarkSims]

I just found this paper about oleic acid and ethanol. Appearently just mixing them together gives good skin penetration. The author seems to think that most of the penetration comes from the oleic acid, and not the ethanol.

http://www.sciencedi...168365995000887

[ClarkSims]

Well I have moved across the country, started a new job and finally got settled in. I mixed up about 100 ml of C60-oo, with about 10 mil of limonene last night.

I put a small amount on my scalp, to check if I would have any irritation from the mixture. This morning I am fine.

I will take a picture once a week or so, and start soaking the scalp every night, and see what happens.

[RedCairo]

Would be interested to see your results.


Regarding DMSO: it will not carry the olive oil. But it would probably carry the fullerenes. If you put it on your skin and then put the OO+F on the skin, would it not be probable that it would pull "some" of the fullerenes through the skin (any that hit the surface in the blend) and leave most of the OO on the outer skin (which is actually ideal since if you don't oil skin awhile after using DMSO you get dehydration effects).

My only concern is this: is there any evidence for what happens when fullerenes go through the BBB? Because "if" "some" of them were taken through by DMSO, they'd cross it. This is really the only thing keeping me from doing this little experiment with the OO+F bottle I ordered from overseas.

PJ

[hav]

Would be interested to see your results.


Regarding DMSO: it will not carry the olive oil. But it would probably carry the fullerenes. If you put it on your skin and then put the OO+F on the skin, would it not be probable that it would pull "some" of the fullerenes through the skin (any that hit the surface in the blend) and leave most of the OO on the outer skin (which is actually ideal since if you don't oil skin awhile after using DMSO you get dehydration effects).

My only concern is this: is there any evidence for what happens when fullerenes go through the BBB? Because "if" "some" of them were taken through by DMSO, they'd cross it. This is really the only thing keeping me from doing this little experiment with the OO+F bottle I ordered from overseas.

PJ

To cross the bbb, a molecule has to be oil soluble and small enough. c60 in olive oil clearly fits the bill, all by itself. I don't think c60 dissolves into dmso very well, if at all. Dmso seems to work better as a trans dermal carrier of water soluble things. Olive oil applied to the skin works pretty well for oil solubles and is pretty good for the skin itself. Dmso, on the other hand, tends to leave my skin dehydrated and pretty wrinkled requiring something like aloe gel to repair it. Perhaps c60/oo might get absorbed more rapidly into dmso-wrinkled skin than normal skin.

Howard

[hav]

Here's a study mentioning using a sonicated combo of dmso and tween80 to achieve a 100 ppm c60 concentration... but it sounds like more of a suspension than a dissolved solution:

Choosing safe dispersing media for C60 fullerenes

Assessment of C(60) nanotoxicity requires a variety of strategies for dispersing it into biological systems. Our objective was to determine organic solvent/surfactant combinations suitable for this purpose. We used Escherichia coli (ATCC# 25254) to determine the cytotoxicity of C(60) in solvents at concentrations up to 100 ppm. In this preliminary study we hypothesized that C(60) toxicity is directly correlated with its degree of dispersion in solution and that more solubilizing solvents induce higher toxicity. Test solvent concentration (1%) and Tween 80 (0.04%) were based on E. coli viability assay. Sonication was used to further enhance C(60) dispersal. The end-point response was measured with viability (in terms of LC(50)) and general metabolic activity (in terms of IC(50)) of E. coli cultures after exposure. The ultimate goal was to select safe dispersing media and enrich the database of C(60) nanotoxicity for NanoQuantitative-Structure-Activity-Relationship (NanoQSAR) applications. LC(50) range was 30 ppm to >400 ppm. IC(50) followed the trend. Among the six solvent combinations, DMSO combined with Tween 80 was the optimum combination for defining a dose-response relationship for assessing its toxicity to E. coli. However, N,N-dimethylformamide has the greatest potential to be a safe solvent for C(60) applications based upon its biocompatibility. Solvent solubility alone could not account for the cytotoxicity observed in this study.

Here's what looks like a pre-publication presentation of the above study with a little additional info in it:
http://ehr.cset.jsum...bstracts 64.pdf

Howard

[nowayout]

My only concern is this: is there any evidence for what happens when fullerenes go through the BBB? Because "if" "some" of them were taken through by DMSO, they'd cross it.

Do scalp capillaries drain via the brain? I don't know anatomy that well but would be surprised if that were the case. Of course there would be some secondary exposure via systemic circulation, but at a lower concentration.

Edited by nowayout, 16 May 2013 - 11:32 PM.

[niner]

My only concern is this: is there any evidence for what happens when fullerenes go through the BBB? Because "if" "some" of them were taken through by DMSO, they'd cross it. This is really the only thing keeping me from doing this little experiment with the OO+F bottle I ordered from overseas.

To cross the bbb, a molecule has to be oil soluble and small enough. c60 in olive oil clearly fits the bill, all by itself.

I don't know. C60-oo is pretty large, heavy, and is charged. There's been erroneous information posted about its molecular weight, which is 720 Daltons just for the c60, plus about that much again for two C18 fatty acid adducts. I think it would be a stretch to get it into the brain. I'd want to see some experimental evidence that it crossed the BBB, with or without dmso. I don't think dmso is particularly magic other than as a transdermal carrier. The only way it could help drugs pass the BBB would be to have a very toxic amount in the bloodstream.

[Jakare]

My only concern is this: is there any evidence for what happens when fullerenes go through the BBB? Because "if" "some" of them were taken through by DMSO, they'd cross it. This is really the only thing keeping me from doing this little experiment with the OO+F bottle I ordered from overseas.

To cross the bbb, a molecule has to be oil soluble and small enough. c60 in olive oil clearly fits the bill, all by itself.

I don't know. C60-oo is pretty large, heavy, and is charged. There's been erroneous information posted about its molecular weight, which is 720 Daltons just for the c60, plus about that much again for two C18 fatty acid adducts. I think it would be a stretch to get it into the brain. I'd want to see some experimental evidence that it crossed the BBB, with or without dmso. I don't think dmso is particularly magic other than as a transdermal carrier. The only way it could help drugs pass the BBB would be to have a very toxic amount in the bloodstream.

What about stacking C60-oo with vinpocetine and MSM. I do not yet understand well the BBB but, wouldn't those supplements increase its permeability in general (and therefore to C60-oo also)?

[hav]

I don't know. C60-oo is pretty large, heavy, and is charged. There's been erroneous information posted about its molecular weight, which is 720 Daltons just for the c60, plus about that much again for two C18 fatty acid adducts. I think it would be a stretch to get it into the brain. I'd want to see some experimental evidence that it crossed the BBB, with or without dmso. I don't think dmso is particularly magic other than as a transdermal carrier. The only way it could help drugs pass the BBB would be to have a very toxic amount in the bloodstream.

Here's a quote directly out of the Baati study... it pertains to the amount of c60 detected in animals that were sacrificed early on:

As C60 contents in lungs and kidneys are likely weaker than those in livers and spleens, we only focused on C60 content in brains because the issue of its translocation to the brain is still a matter of debate [25,43]. Whereas C60 particles were not detected in the brain after intratracheal instillation [43], 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 blood-brain barrier [25].

Howard

Edited by hav, 19 May 2013 - 05:08 PM.

[ClarkSims]

Sorry I have taken so long to respond. I have been working really long hours. I would like to post before pics, and current pics. I think there has been little improvement in the gray hair. I can say for sure, that there are no individual gray hairs, which are black at the root and grey further out. So any improvement, would be from new hair follicles coming out of dormancy, and existing gray hair follicles going into dormancy.

*sigh* I should have taken more pictures, and taken pictures around my lips. I think there has been an improvement in the fraction of dark hair, but I don't have the pics, or the hair counts to prove it. Alas, the my clients ask to much of me. I really need to just make a lot of money, and get rid of my clients.

[ClarkSims]

As C60 contents in lungs and kidneys are likely weaker than those in livers and spleens, we only focused on C60 content in brains because the issue of its translocation to the brain is still a matter of debate [25,43]. Whereas C60 particles were not detected in the brain after intratracheal instillation [43], 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 blood-brain barrier [25].

Since the c60 is adducted to the fatty acid molecules, and the fatty acid molecules are used in every membrane in the body, it makes sense that it would cross the blood brain barrier.

I can't remember the name of the packages of small fatty acids, and wikipedia hasn't played nice this morning. I remember that the duodenum puts long chain fatty acids into small packages, which then flow through out the body. IIRC, they flow primarly through the lymph on the the first pass. The packages are broken apart, and the liver cleans up the fragments.

I don't understand how lymph flows through the brain. I wonder if the brain gets its fatty acids from the lymph?

[niner]

I don't know. C60-oo is pretty large, heavy, and is charged. There's been erroneous information posted about its molecular weight, which is 720 Daltons just for the c60, plus about that much again for two C18 fatty acid adducts. I think it would be a stretch to get it into the brain. I'd want to see some experimental evidence that it crossed the BBB, with or without dmso. I don't think dmso is particularly magic other than as a transdermal carrier. The only way it could help drugs pass the BBB would be to have a very toxic amount in the bloodstream.

Here's a quote directly out of the Baati study... it pertains to the amount of c60 detected in animals that were sacrificed early on:

As C60 contents in lungs and kidneys are likely weaker than those in livers and spleens, we only focused on C60 content in brains because the issue of its translocation to the brain is still a matter of debate [25,43]. Whereas C60 particles were not detected in the brain after intratracheal instillation [43], 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 blood-brain barrier [25].

Since the c60 is adducted to the fatty acid molecules, and the fatty acid molecules are used in every membrane in the body, it makes sense that it would cross the blood brain barrier.

Thanks Howard and Clark. Baati does provide evidence that c60-oo gets through the BBB. And Clark is right- there needs to be a way for fatty acids to get in, and in fact there are fatty acid transporters for that very purpose. The c60 is probably hitching a ride on a fatty acid transporter.

[ClarkSims]

Well after 4 months of treating my scalp with OO-C60 and limonene, I don't see any results :-( Perhaps 4 months isn't long enough? Perhaps I should try some other anti oxidants also?
Here are some photos.

sorry for the long load times. The pictures are all high res.

http://lab.homelinux.org/hair/