2523 replies to this topic

[Learner056]

Beautiful post.  

 

Turnbuckle you wrote: 

the endogenous proliferation of stem cells should provide the most direct and comprehensive approach to restoring cellular maintenance and extending lifespan.

 

You introduced to us some good concepts earlier.  I presume you mean here, "endogenous proliferation of SCs that is cognizant of symmetric replication (i.e. self-renewal aspect)?  so as to prevent their ultimate depletion .. as that would occur if self-renewal is not factored in.   

 

 

You wrote: 

With time, cells undergo epigenetic mutations (epimutations) that degrade cellular performance. These occur far more frequently than with the underlying genetic code, and there is no repair mechanism except replacement.  Thus replacing old epimutated cells with new cells derived from SCs having de novo methylation will improve the performance of organs and the body as a whole. 

 

You view epimutations as detrimental.  I find this concept difficult, as when I as impartial observer view epiMutations, I see a savior.  These epiMutations could be an evolutionary advantage to keep us alive (instead of us succumbing rapidly to our fundamental genetic defects).   Case in point: John has a major ERCC5 defect, i.e. he has a limited DNA repair capacity.  With an altered lifestyle John has managed to stay very healthy.  John has significant epiMutations - but ironically most seem to be associated with longevity (low igf, reduced heme synthesis, reduced protein turnover etc).  So with all epiMutations erased, that puts John back at a evolutionary dis-advantage, where his new cells would be subjected to the limited damage repair capacity, but with OxPhos this time running at full capacity, the future mutations that will quickly accumulate may not be as friendly this time?

 

You wrote:

replaced by cells derived from proliferated SCs of part A

 

I did not catch, what is Part A and Part B. 

 

You wrote: 

Telomerase promoters like resveratrol should definitely be avoided

 

I appreciate the guidance to follow the protocol strictly.  But for broader knowledge, what is the rationale of not using a Telomerase promoter here?  

Would it be ok to use it with your other Mitochondrial protocol i.e. without C60 (a little passionate about Resveratrol here, it was first supplement I ever tried and with great positive effects)

 

 

Edited by Learner056, 07 September 2022 - 07:57 PM.

[Turnbuckle]

 

You view epimutations as detrimental.  I find this concept difficult, as when I as impartial observer view epiMutations, I see a savior.  These epiMutations could be an evolutionary advantage to keep us alive (instead of us succumbing rapidly to our fundamental genetic defects).   Case in point: John has a pathogenic ERCC5 defect, i.e. he has a limited DNA repair capacity.  With a crafted lifestyle John has managed to stay very healthy.  John has significant epiMutations - but ironically they seem to be associated with longevity (low igf, reduced heme synthesis, reduced protein turnover etc).  So if epiMutations are erased, that puts John back at a evolutionary dis-advantage, where his new cells would be subjected to the limited damage repair capacity, but with OxPhos this time at full capacity, the future mutations that accumulate this time may not be friendly?

 

 

 

1. Unlike with genetic mutations of reproductive cells, epimutations are not inherited by the next generation, and there is no evolution within a single individual.

2. Epimutations are not erased by this process. Cells that are telomerically old are replaced with cells with de novo programming -- a freshly minted epigenetic code. 

 

Think of the genetic code as being the piano and the epigenetic code as the sheet music. There are 200 different sheets, one each for the 200 different cell types. Random epigenetic mutations corrupt the notes like a bad Xerox machine, so that what starts off as a liver cell playing Rachmaninoff at the age of 6 ends up being chop sticks when you reach 100.

 

replaced by cells derived from proliferated SCs of part A

 

I did not catch, what is Part A and Part B. 

 

 

Dang. When I copy pasted this bit from Word, the auto numbering changed from letters to digits. 1-3 in that section should have been A-C. So I'll repost this after all the errors show up thanks to sharp readers.

 

You introduced to us some good concepts earlier.  I presume you mean here, "endogenous proliferation of SCs that is cognizant of symmetric replication (i.e. self-renewal aspect)?  so as to prevent their ultimate depletion .. as that would occur if self-renewal is not factored in.  

 

 

Proliferation is symmetric.

Edited by Turnbuckle, 07 September 2022 - 08:01 PM.

[kurt9]

So Fisetin really does work as a senolytic. I notice from your graph that curcumin does as well.

[Learner056]

But you taught us earlier that Proliferation of stem cells can be: symmetric "OR" assymetric. 

Where symmetric division aka prolif = 2 daughter stem cells

Where assymetric division aka prolif = 1 daughter stem cell + 1 differentiated cell. 

 

While here onwards I can be wrong in verbiage (non biology major here, ancient) but the essence of what you said earlier was: that stem cells divide in some 80:20 ratio (80% chance of asymmetric division vs 20% chance symmetric division).  And you said Fusion that helps stem proliferation (but with its default 80:20 division ratio on its own may not be sufficient) but when combined with C60 (blocking MMP pore etc) helps skew this ratio more towards symmetric division, e.g. it will now be instead of the paltry 80:20, it may become 70:30 or lets say more ambitiously 40:60 i.e. in anycase whatever the numbers, the combination helps beef up symmetric division aka proliferation. 

 

 

Turnbuckle you wrote:

Proliferation is symmetric.

 

In response to:

You introduced to us some good concepts earlier.  I presume you mean here, "endogenous proliferation of SCs that is cognizant of symmetric replication (i.e. self-renewal aspect)?  so as to prevent their ultimate depletion .. as that would occur if self-renewal is not factored in.

 

Edited by Learner056, 08 September 2022 - 01:16 AM.

[Turnbuckle]

 

But you taught us earlier that Proliferation of stem cells can be: symmetric "OR" assymetric. 

 

 

 

Really? I hope I didn't, as proliferation is a symmetric process of division that yields two new stem cells. Differentiation is a process of asymmetric division that yields one new stem cell and one cell that  isn't a stem cell.

[Learner056]

Must have been I that mistook. One critical concept (below), I struggle, appreciate if you or someone here can articulate it to me: 

 

What does the fuel source being used (e.g. glycolysis vs aerobic glycolysis vs OxPhos) tell us about: fission / apoptosis (for cell replacement) / fusion / biogenesis (for stem cell proliferation)?. 

 

"Azithromycin treatment of human fibroblasts was indeed sufficient to strongly induce both aerobic glycolysis and autophagy. However, the effects of Azithromycin on mitochondrial oxygen consumption rates (OCR) were bi-phasic, showing inhibitory activity at 50 μM and stimulatory activity at 100 μM. These autophagic/metabolic changes induced by Azithromycin could mechanistically explain its senolytic activity"

Edited by Learner056, 08 September 2022 - 06:12 PM.

[Termophilus]

Tried the protocol for a second time in a week, I've taken all the supplements except sunflower lecithin, sam-e and dihydromyricetin.

The first time I've noticed something about my metabolism going up, and a little bit anxious, but not too much.

 

After two days of fission I've tried a second round and this time the change in metabolism was quite strong.

I've taken the supplements in the evening, around 7 PM, and I after going to sleep at 2 AM, maybe after three hour of sleep I woke up because I felt very hot, and somewhat anxious, expecially I felt it in the heart region, my feelings were that my heartbeat was irregular, but I didn't checked with some measurements.

 

Then I've taken a cold shower to cool down, but I didn't felt I was going to sleep again soon, because of the anxiousness, so I drink some milk with honey and whatched something on the pc.

 

After one hour maybe I felt good again and I was succesful sleeping. My dreams were quite wild and vivid after.

 

I think I will stay with only one dose x week of C60 in the future, because I understand this is the supplement which is involved in mitochondrial metabolism, and I've found some research about the cardiovascular system particularly targeted by this.

Any advice for safety? I don't have the time and the skill to study the lecterature, maybe I'm doing it too recklessy and I need some advice, which could be useful even for other who want to experiment with it without knowing all the biological mechanism involved. 

 

 

[Turnbuckle]

Tried the protocol for a second time in a week, I've taken all the supplements except sunflower lecithin, sam-e and dihydromyricetin.

The first time I've noticed something about my metabolism going up, and a little bit anxious, but not too much.

 

 

 

C60 hypothetically blocks proton leakage in SC mitochondria and thus wakes them up. Same thing for somatic cells, but the effect is 5-10 times less as there are that many fewer UCP2 pores compared to SCs. So it's a side effect, and not directly connected to what's happening with stem cells.

Edited by Turnbuckle, 08 September 2022 - 07:16 PM.

[Confectman]

@Turnbuckle You mentioned not to combine stearic acid with any supplements. How about mitochondria driving peptides like HNG, SS31, Mots-c ? are they helpful or should I stop them during protocol days?

 

 

[Turnbuckle]

@Turnbuckle You mentioned not to combine stearic acid with any supplements. How about mitochondria driving peptides like HNG, SS31, Mots-c ? are they helpful or should I stop them during protocol days?

 

 

I don't recall saying that. You can combine stearic acid with DHM, for instance. Still, adding random things to the protocol is not a good idea unless you have an excellent reason. The ones you mention seem unlikely to cause a problem, but why take the chance if you're going to use fusion/C60 just once a week? And if you are using these peptides to improve mito performance, you might try my mito protocol first. In which case you might not need these peptides any more.

Edited by Turnbuckle, 09 September 2022 - 04:22 PM.

[Repack Racing]

I've taken the supplements in the evening, around 7 PM

 

Other have suggested taking in the evening, but I strongly recommend doing these protocols in the AM - especially if you have any existing sleep issues.  That gives your body time to process the supplements and put them to use. 

[Repack Racing]

@Turnbuckle You mentioned not to combine stearic acid with any supplements. How about mitochondria driving peptides like HNG, SS31, Mots-c ? are they helpful or should I stop them during protocol days?

 

I have run all of these alongside the protocol.  I find Mots-c to be the only one that really benefits me while doing the protocol.  SS-31 has some very good studies, but I personally do not notice anything. It's possible there isn't any added benefit over the protocol or that it's doing something that I cannot perceive.  I personally avoid HNG due to a bad experience with it. 
 

[Repack Racing]

deleted

Edited by ildr, 09 September 2022 - 08:23 PM.

[Kelvin]

Here is my C60 protocol.

 

It is similar to Turnbuckle's but it uses capsules instead of blending them, except for olive oil C60 which I take a teaspoon of.

 

Also, I do not make the brownies.

 

My C60 bottle is double wrapped in aluminum foil and put in the freezer to prevent it from being exposed to light (which degrades C60's stability and can make it toxic) and to preserve C60's molecular structure with cold temperatures.  

 

I only take it out of the freezer the night before I use it to warm it up and place it (still wrapped in aluminum) in a dark closet overnight.  After I briefly take a teaspoon of olive oil I double wrap the bottle of C60 again with aluminum foil and place it back in the freezer.

 

I replace the bottle every 6 months (even if it is still full) and buy a new one to ensure the purity of the C60.

 

My version of the protocol spreads out the different components across 20-30 minutes to make it easier to take so many supplements -

 

 

***** 0 hours *****

 

4 grams Dihydromyricetin (I might drop this to 2 or 3 grams because I find 4 grams to be draining)

2 grams Sunflower Lecithin (To prevent blood pressure from rising because of the fusion agents)

3 grams AAKG 

 

 

 

***** 10 to 20 minutes later *****

 

1 gram Liposomal Glutathione 

1.2 grams AKG

100 mgs Sulforaphane

--------> 500 mgs SAM-e   (Note - ONLY use SAM-e if you are NOT using 5-HTP because SAM-e and 5-HTP are not good if mixed together) 

 

 

--------> 500 or 1,000 mgs  TMG   (Note - ONLY use TMG if you are using 5-HTP) 

-------->  100 mgs HTP-5 (Note - This is optional and could be done once after every 3 to 4 cycles.  Do NOT take more than 100 mgs of 5-HTP in a 24 hour period.  Only use 5-HTP with TMG and NEVER with SAM-e because SAM-e and 5-HTP are not good if mixed together) 

 

 

***** 10 minutes later *****

 

--------> 500 mgs Astragalus Root Extract (Use astragalus root once every 10th cycle)

 

2 grams Methionine

2 grams Lysine

 

3 mgs C60 in extra virgin olive oil (Taken AFTER having every other supplement)

 

 

 

*** Take the same amino acids (2 grams methionine and 2 grams lysine) whenever you feel sleepy, or, every 1 - 3 hours for that day and the next three days to keep feeding your stem cells.   

 

Remember to have LOTS of methionine and lysine supplements available because you will feel sleepy if you don't feed your stem cells with these two amino acids.  

 

I recommend having 500 capsules of lysine and 500 capsules of methionine with 500 mgs per 1 capsule on hand whenever doing the C60 cycle.

 

 

 

*** Senolytics.  The day after taking C60 take senolytics and fission promoters once in the morning for 2 days, but not more than 3 days.  One could also drop senolytics if you aren't seeing benefits and just use AAKG, NR, and NMN for fission promotion.

 

 

3 grams AAKG 

500 mgs Fisetin (Senolytic.  Can be reduced to 100 mgs if one stops feeling an effect from them)

500 mgs Quercetin (Senolytic)

1 gram Curcumin (Senolytic)

100 mgs Apigenin (Senolytic)

300 mgs NRcl (Nicotinamide Riboside Chloride)

750 mgs NMN 

 

 

Finally, consider taking SAM-e or TMG every 3 cycles to replenish your body's methyl stores which are probably depleted from taking so many demethylating AKG supplements.

Edited by Kelvin, 09 September 2022 - 09:59 PM.

[Kelvin]

deleted by author

Edited by Kelvin, 09 September 2022 - 10:13 PM.

[Learner056]

Super-informative.  Turnbuckle deserves thanks that does not require saying, but yours going through the concepts below (that I, who is well past my age, but many others here - young people you all are, will benefit) it speaks to your character VERY highly. Thank you Kelvin. 

 

 

Here is my C60 protocol. 

 

SAM-e and 5-HTP are not good if mixed together

 

Remember to have LOTS of methionine and lysine supplements available because you will feel sleepy if you don't feed your stem cells with these two amino acids.  

 

Finally, consider taking SAM-e or TMG every 3 cycles to replenish your body's methyl stores which are probably depleted from taking so many demethylating AKG supplements.

 

 

[timedilation]

I'm curious to learn more about the demethylation aspects of this protocol.  Does anyone have interesting studies on the effects of demethylators on epigenetic age?  Apologies if anything has already been posted in this thread.

 

 

I use both AKG (already dissolved by the manufacturer) and AAKG. The first is very fast acting (more important during fusion) and the other somewhat slower (more important during fission). Calcium AKG is expected to be even slower and may be used instead of AAKG. Other AKG salts or AKG conjugates may be used.

 

Do we know that rate of activity is the only real difference among Ca-AKG, AKG, and AAKG?  Is there any chance the total demethylation activity is reduced in AAKG or Ca-AKG?

 

Also, are there any other substances that could potentially complement AKG in this task?

[QuestforLife]

I'm curious to learn more about the demethylation aspects of this protocol. Does anyone have interesting studies on the effects of demethylators on epigenetic age? Apologies if anything has already been posted in this thread.

Do we know that rate of activity is the only real difference among Ca-AKG, AKG, and AAKG? Is there any chance the total demethylation activity is reduced in AAKG or Ca-AKG?

Also, are there any other substances that could potentially complement AKG in this task?

See this paper:
Demidenko O, Barardo D, Budovskii V, Finnemore R, Palmer FR, Kennedy BK, Budovskaya YV. Rejuvant®, a potential life-extending compound formulation with alpha-ketoglutarate and vitamins, conferred an average 8 year reduction in biological aging, after an average of 7 months of use, in the TruAge DNA methylation test. Aging (Albany NY). 2021 Nov 30;13(22):24485-24499. doi: 10.18632/aging.203736. Epub 2021 Nov 30. PMID: 34847066; PMCID: PMC8660611.

Note, I've used both kirkman AKG (Mg and Ca salt mix) and separately arginine-AKG starting from the Kirkman supplement in April 2020, substituting for AAKG later, and both reverse my methylation age by 5-7 years. So it is not necessary to take the Rejuvenant product.

I've also taken long (3 month) breaks, started again for a week or two, and then tested, and maintained my lower epigenetic age. But it took 6 months to get the effect (no reduction@3 months) when I first started. So it appears AKG declines slowly with age, and once topped up stays that way for a while ; probably a function of mitophaghy - as described in this paper:

Vazquez-Martin A, Van den Haute C, Cufí S, Corominas-Faja B, Cuyàs E, Lopez-Bonet E, Rodriguez-Gallego E, Fernández-Arroyo S, Joven J, Baekelandt V, Menendez JA. Mitophagy-driven mitochondrial rejuvenation regulates stem cell fate. Aging (Albany NY). 2016 Jul;8(7):1330-52. doi: 10.18632/aging.100976. PMID: 27295498; PMCID: PMC4993334.

There's a whole AKG thread somewhere, but can't find it on my phone.

[Empiricus]

Other have suggested taking in the evening, but I strongly recommend doing these protocols in the AM - especially if you have any existing sleep issues.  That gives your body time to process the supplements and put them to use. 

 

Starting the protocol in the evening reduces the likelihood of any (hypothetical) dangerous interaction between sunlight hitting your skin and the c60 in your blood.  It's conceivable that even brief exposures to sunlight might be quite hazardous in the hours after you consume c60.

 

Since we can't be sure how long c60 circulates, it might be wise to avoid sunshine for longer than overnight. One possible explanation for my miserable experience beginning the protocol in early July is that I took a walk in bright sunlight about 8 hours after taking the c60.  

 

Of course, if it's December and you live in Utqiagvik, it shouldn't make any difference what time you start the protocol.  

Edited by Empiricus, 11 September 2022 - 10:34 AM.

[trying2survive]

new study Mitochondrial dynamics maintain muscle stem cell regenerative competence throughout adult life by regulating metabolism and mitophagy talks about mito fissionand stem cells etc. Interestingly, DCA (not FDA approved) seems to help, "We confirmed that DCA treatment significantly increased OCR and mitochondrial ATP production rates in SCs (Figure S5D). Of note, daily DCA treatment (for 4 days) of Drp1ΔPax7ER mice increased the levels of proliferating SCs (Ki67+YFP+) in injured muscles, as shown in tissue cryosections (Figure 5F), and rescued the defective muscle regeneration, as shown by increased numbers and sizes of newly formed YFP+ (or eMHC+) myofibers in muscle after injury."

[Learner056]

I have been on unsweetened dark chocolate (100% cocoa) for Fusion days with C60 which works really well for me, but now thinking to upgrade.  I have some Myricetin in stock that I should finish before I switch to DHM for long term.  Would there be some rough dosing equivalence between Myricetin vs DHM? like how much 1g of DHM = xx mg Myricetin?

[Learner056]

Strange, my experience is a little opposite (not that I have tried being in sun for extended period), but I consistently experience single "topical dose" C60 efficiently penetrating facial skin and scalp and being in systemic circulation for days.  I feel post-application it evidentially soaking in arm muscles (3-4 hrs later, peaking at 24 hrs, and then remaining and fading over a week).  On the contrary, C60 molar mass is high enough to prevent this all from occurring?  Could this be even real?

 

interaction between sunlight hitting your skin and the c60 in your blood

 

Edited by Learner056, 12 September 2022 - 04:12 AM.

[Turnbuckle]

 On the contrary, C60 molar mass is high enough to prevent this all from occurring?  Could this be even real?

 

Substances that are intended to cross the skin must have a molecular weight below 1000 daltons. The size is probably not the limiting factor but with increasing size the chemical structure becomes more complex, the partitioning behaviour changes and penetration is therefore reduced. https://www2.mst.dk/...l/kap09_eng.htm

 

 

C60 is 720 daltons. 

Edited by Turnbuckle, 12 September 2022 - 10:08 AM.

[Advocatus Diaboli]

From post 2003:

 

"Substances that are intended to cross the skin must have a molecular weight below 1000 daltons. The size is probably not the limiting factor but with increasing size the chemical structure becomes more complex, the partitioning behaviour changes and penetration is therefore reduced. https://www2.mst.dk/...l/kap09_eng.htm"

 

And, on the other hand, there is the 500 dalton rule.

[QuestforLife]

Glutamine, Leucine and Arginine (with fasting from other AAs) may be of interest to increase mitochondrial fusion:

 

Mitochondrial hyperfusion via metabolic sensing of regulatory amino acids (1)

 

This is a very interesting paper on mitochondrial hyperfusion triggered by the consumption of certain amino acids (AAs) specifically: glutamine, leucine, arginine.

Firstly, a mild fusion state is triggered by cell AA starvation and then severe hyperfusion is induced by addition of the three AAs mentioned. Maximum effect is from 4-8 hours.

Effect requires absence of other AAs, which block fusion. But glucose only partially blocks fusion.

 

So a human could overnight fast, then take Glut+leuc+argin to get these effects (if desired). Fasting should continue at least 4 hours, with lunch being carb (not protein) based.

Mechanism of action was examined with the two most effective AAs of the three (glut+leuc) acting through Krebs cycle and downstream, the creation of GTP, which upregulates fusion protein mfn1. One wonders if direct supplementation of the guanosine nucleoside would be easier (if possible)?

 

Of further note:

1. dGTP concentrations increase speed of the telomerase protein activity, see my discussion link(2), paper ref (3). Therefore upregulation of GTP by this mechanism could benefit (stem) cells that have low telomerase levels by increasing how many base pairs telomerase can add to the telomere during s-phase.

2. Alpha ketoglutarate has been shown to reverse methylation age, and likely overlaps with the AA pathway described above, see my discussion link(4), paper ref (5). So leucine and glutamine consumption would be expected to do the same (my prediction).

3. As a corollary of the above, depletion of glutamine, leucine and arginine would be expected to increase methylation age (again, my prediction), and might explain negative effect of telomerase activators on methylation-age, mediated via increased cellular division (which is known to use up glutamine, see my previous discussion (6)).

4. The advantages of mitochondrial hyperfusion for Turnbuckle's 'stem cell protocol' are obvious (7).

 

References:

(1)Mitochondrial hyperfusion via metabolic sensing of regulatory amino acids, Abdullah et al., 2022, Cell Reports 40, 111198 August 16, 2022 https://doi.org/10.1...rep.2022.111198

(2) Chen Y, Podlevsky JD, Logeswaran D, Chen JJ. A single nucleotide incorporation step limits human telomerase repeat addition activity. EMBO J. 2018 Mar 15;37(6):e97953.https://doi:10.15252/embj.201797953, Epub 2018 Feb 12. PMID: 29440226; PMCID: PMC5852417.

(3)Nucleotides (specifically guanine) for elongation of telomeres: eat Anchovies and Herring!
https://www.longecit...-15#entry904277

(4)Plan to reduce both telomere and methylation age:
https://www.longecit...e-9#entry895170
No improvement in methylation age from 3 months of AKG:
https://www.longecit...-10#entry896760
Improvement in methylation age from 6 months of AKG:
https://www.longecit...-13#entry899822

(5)Demidenko O, Barardo D, Budovskii V, Finnemore R, Palmer FR, Kennedy BK, Budovskaya YV. Rejuvant®, a potential life-extending compound formulation with alpha-ketoglutarate and vitamins, conferred an average 8 year reduction in biological aging, after an average of 7 months of use, in the TruAge DNA methylation test. Aging (Albany NY). 2021 Nov 30;13(22):24485-24499. https://doi:10.18632/aging
203736 Epub 2021 Nov 30. PMID: 34847066; PMCID: PMC8660611.

(6)Glutamine depletion from telomerase activators? https://www.longecit...-24#entry911858

(7)Turnbuckle's protocol: https://www.longecit...-24#entry911858

 

 

 

 

[Turnbuckle]

From post 2003:

 

"Substances that are intended to cross the skin must have a molecular weight below 1000 daltons. The size is probably not the limiting factor but with increasing size the chemical structure becomes more complex, the partitioning behaviour changes and penetration is therefore reduced. https://www2.mst.dk/...l/kap09_eng.htm"

 

And, on the other hand, there is the 500 dalton rule.

 

 

For every rule of thumb, there are exceptions--

 

Quantification of the number of hair follicles in treated vs.

nontreated animals revealed a statistically significant

increase in fullerene-treated mice when given intradermally

(Figure 4, A) or topically (Figure 4, B). Thus, fullerenes not

only accelerate the rate at which hair shafts grow but also

induce de novo synthesis of new hair follicles.

https://pubmed.ncbi....h.gov/19223242/

 

 

[Advocatus Diaboli]

Turnbuckle, unfortunately the particular fullerenes used by the investigators in the study you cite in post #2006 aren't identified (full study is here). And, as you know, there are a huge  number of fullerenes. So, no conclusion about the skin permeability of C60 can be drawn from your cited reference.

 

The reference you cite in post #2003, which gives the 1,000 dalton value has, at the end of the paragraph, a reference to (Guy et al., 1987).

 

Guy, et al. write:

 

"Firstly, the drug will probably have a molecular weight less than approximately 1000 daltons. There is nothing clear-cut about the number 1000, it merely reflects the fact that most simple drug molecules have molecular weights which fall below this figure."

 

The reasoning presented by Guy, et al. is consistent with the reasoning given in the reference I cited in post #2004:

 

"1) virtually all common contact allergens are under 500 Dalton, larger molecules are not known as contact sensitizers. They cannot penetrate and thus cannot act as allergens in man; 2) the most commonly used pharmacological agents applied in topical dermatotherapy are all under 500 Dalton; 3) all known topical drugs used in transdermal drug-delivery systems are under 500 Dalton. In addition, clinical experience with topical agents such as cyclosporine, tacrolimus and ascomycins gives further arguments for the reality of the 500 Dalton rule."

 

 

[Turnbuckle]

Turnbuckle, unfortunately the particular fullerenes used by the investigators in the study you cite in post #2006 aren't identified (full study is here). And, as you know, there are a huge  number of fullerenes. So, no conclusion about the skin permeability of C60 can be drawn from your cited reference.

 

The reference you cite in post #2003, which gives the 1,000 dalton value has, at the end of the paragraph, a reference to (Guy et al., 1987).

 

Guy, et al. write:

 

"Firstly, the drug will probably have a molecular weight less than approximately 1000 daltons. There is nothing clear-cut about the number 1000, it merely reflects the fact that most simple drug molecules have molecular weights which fall below this figure."

 

The reasoning presented by Guy, et al. is consistent with the reasoning given in the reference I cited in post #2004:

 

"1) virtually all common contact allergens are under 500 Dalton, larger molecules are not known as contact sensitizers. They cannot penetrate and thus cannot act as allergens in man; 2) the most commonly used pharmacological agents applied in topical dermatotherapy are all under 500 Dalton; 3) all known topical drugs used in transdermal drug-delivery systems are under 500 Dalton. In addition, clinical experience with topical agents such as cyclosporine, tacrolimus and ascomycins gives further arguments for the reality of the 500 Dalton rule."

 

 

Yes, I'd seen the paper. They are using C70 because this allows them to graft adducts to the polar regions of the molecule. Their goal is to get a patented product they can sell. But nevertheless, it shows that the 500 Dalton rule has its exceptions, as these derivatives are substantially heavier than C60. 

 

Another paper showed that C60 can penetrate pig skin--

 

The application of fullerene C60 for transdermal delivery was assessed and validated through in vitro tests using pig skin as a model. Fullerene C60 was found to be able to cross the intact skin after its dispersion in a solution of fatty acids (IPM with 20% TRC).   PMID: 32624818

 

[Advocatus Diaboli]

In post #2006, Turnbuckle writes: "For every rule of thumb, there are exceptions--"

 

Agreed. As I noted in 2017 in post #26 of the "Methylene blue homemade skin cream?" thread:

 

"Yeah, some people take the "500 Dalton rule" as gospel, not having considered the exceptions such as the "needle" effect.".

 

--In response to Aconita's post #25 in which he coins "needle" effect to describe a condition in which a high-dalton molecule, such as hyaluronic acid, will pass through the skin whereas a more spherically shaped, and equal-weighted, molecule would not be expected to.

 

"Another paper showed that C60 can penetrate pig skin--"

 

Circe's victims on Aeaea might have been pleased to learn that the emollients that they used as men probably would have still worked after their transmogrification.

[FWP]

Yes.

The body replaces some 300 billion cells a day, much of that via TACs (transit amplifying cells that are intermediate between SCs and somatic cells). TACs appear to be the locus of epigenetic aging in many tissues, so replacing TACs via SCs more frequently will lower the average epigenetic age of those tissues. A good number of those 300 billion replaced cells are physically lost (epidermal and epithelial cells, for example), but the others require apoptosis. Fission is required for apoptosis, but the fusion state is sticky, so you can speed things up with a fission supplement to banish fusion once it's not needed. Senolytics can take care of those senescent cells that that have become resistant to apoptosis.

Bottom line, an occasional C60/EVOO + mito fusion treatment will decrease epigenetic age (and can be expected to increase longevity as was seen in the Baati experiment). Adding a demethylase promoter will improve the results. Adding fission + apoptosis promoters (senolytics) with further improve the results.

The Baati experiment likely had C60/EVOO + mito fusion, but didn't appreciate what they had. They fasted rats overnight in their toxicity study, but failed to say anything about feeding with the subsequent longevity study. Overnight fasting would have been sufficient to produce a fasting-induced state of fusion, as rodents have a metabolic rate 6 times that of humans. But they made such a good case for C60 operating via an anti-oxidant mechanism that no one considered the mechanism might be completely different. And thus one attempt at replication has failed after another.

As I understand fasting triggers mito fusion and the first part of the protocol also triggers fusion. What I do not get is fasting triggers autophagy and apoptosis which we achieve with the second part of the protocol. So when I hear fasting is used in the c60 study I always thought it is the second part of the protocol achieving this instead of the first part. Still quite confusing for me as I intuitively feel going into a fasting state using nicotinamide rather than stearic acid. Just trying to wrap my head around this