2523 replies to this topic

[Empiricus]

Very Interesting thread, I've read some but not all (75 pages!) of it.  Too bad that TB isn't still here; I value everyone's input but his especially.  

 

About the protocol, why not simplify it,  at least at first?  

 

So, say:  a 24 hour fast with light cardio -> pool your stem cells -> C60 5ml.  Once a week?

24 hours fast may be not be long enough.  

 

Probably safest to first add fasting to protocol, instead of eliminating everything that several have found actually seems to work. 

[Kelvin]

Very Interesting thread, I've read some but not all (75 pages!) of it. Too bad that TB isn't still here; I value everyone's input but his especially.

About the protocol, why not simplify it, at least at first?

So, say: a 24 hour fast with light cardio -> pool your stem cells -> C60 5ml. Once a week?

Humans, unlike rats, would need to fast longer than a day to trigger fusion.

Besides, why induce fusion with fasting which most people would find uncomfortable to do when mitofusion can be triggered almost immediately by just taking fusion promoting capsules like DHM, sulforaphane, or MSG?

Edited by Kelvin, 05 January 2023 - 10:27 PM.

[Kelvin]

I’ve done more research into how UCP2 inhibition might damage cancer cells by making them lose control of their oxidation suppression mechanisms. I also looked into Fisetin’s effect on oxidation and cancer cells.

I wanted to see if the C60 protocol could be modified into an anti cancer regimen.

Based on what I found at these links I think the protocol as it is should be damaging to cancer cells, with only a slight tweak needed to enhance it.

Mitochondrial uncoupling protein 2 and pancreatic cancer: A new potential target therapy

https://www.ncbi.nlm...les/PMC4363752/

Among the regulated genes, UCP2 expression was predominantly higher in rho0 cells suggesting that UCP2 may inhibit ROS accumulation and protect the cells from excessive ROS production induced by mitochondrial defects linked to Warburg effect. In this respect, UCP2 may function as a potential diagnostic marker of cancer associated with the Warburg effect[42]. In addition to its antioxidant role, UCP2 acts as a direct metabolic regulator contributing to the Warburg phenotype. Indeed, as schematically reported in Figure ​Figure2,2, UCP2 has been proposed to function as a uniporter for pyruvate, which promotes pyruvate efflux from mitochondria, restricts mitochondrial respiration, and increases the rate of glycolysis in cancer cells[43]. Furthermore, UCP2 catalyzes the exchange of intramitochondrial C4 metabolites for cytosolic phosphate by an H+-assisted mechanism, which is stimulated by both the electrical potential (negative inside) and pH gradient (acidic outside) existing across the inner mitochondrial membrane of respiring cells[44]. In particular, by exporting oxaloacetate and related C4 compounds from mitochondria, UCP2 negatively controls the oxidation of acetyl-CoA-producing substrates via the Krebs cycle, thus lowering the redox pressure on the mitochondrial respiratory chain, the ATP: ADP ratio, and ROS production. Notably, the mitochondrial concentration of oxaloacetate is usually very low, and its availability regulates the entry of acetyl-CoA into the Krebs cycle. Thus, UCP2 prevents mitochondrial glucose oxidation and favors a higher glucose utilization by aerobic glycolysis. In this context, our research group further confirmed the pro-glycolytic effect of UCP2 demonstrating for the first time that UCP2 can stabilize the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in the cytoplasm of cancer cells[45]. Accordingly, in response to oxidative stress, GAPDH has been demonstrated to undergo protein oxidation of redox-sensitive cysteine residues that stimulates its translocation to cell nuclei[46], where the enzyme favors transcriptional induction of cell death-related genes[47,48]. Thus, the antioxidant effect of UCP2 can inhibit both GAPDH oxidation and nuclear translocation supporting the glycolytic flux and preventing cancer cells from stimulating cell death mechanisms (Figure ​(Figure22).

…

A careful analysis of the recent scientific literature concerning the role of UCP2 in tumor development reveals that UCP2 and cancer may have a double relationship. Indeed, a dual regulation of UCP2 expression, depending on the stages of cancer development, has been observed in many tumor types. A number of studies have established the key role that UCP2 has in tumorigenesis and in chemoresistance. The generally accepted thesis envisages that, during the first stages of tumorigenesis, UCP2 is repressed to allow ROS accumulation and genomic instability, while it is triggered or over-expressed in the following stages of cancer development, determining chemoresistance and tumor aggressiveness by defending cancer cells from apoptosis through the negative regulation of mitochondrial ROS production (Figure ​(Figure33)[49-51]. Accordingly, UCP2-null mice have a predisposition for enhanced tumorigenesis in the proximal colon, providing the first in vivo confirmation of a link between mitochondrial uncoupling proteins and cancer[52], while highly expressed UCP2 is associated with metastatic colon cancer and tumor aggressiveness[53]. The dual and opposite regulation of UCP2 expression in various stages of tumor development has also been demonstrated in breast cancer. In this system, the repression of UCPs by estrogens, a major risk factor for breast cancer initiation, may play a key role in estrogen-induced breast carcinogenesis[54]. On the contrary, the enhanced expression of UCP2 has been correlated to breast cancer progression. Indeed, a significant correlation between UCP2 levels and tumor grade-associated functional phenotypes has been found in a large number of breast cancer patients (n = 234)[55]. Concerning PC, some studies have shown that the protein level of UCP2 is significantly higher in human PC samples than in the adjacent normal tissues, suggesting that UCP2 may promote tumor growth in this tumor type[56]. An extensive study on Oncomine data sets addressed to analyze the UCP2 expression level in a number of cancer types, including pancreatic cancer, has revealed that UCP2 is over-expressed in ovarian, bladder, esophageal, testicular, kidney, colorectal, lung, breast, leukemia, prostate, as well as pancreas cancers[42]. This study has concluded that UCP2 over-expression is a general phenomenon linked to the progression of human cancers.

Along this line of evidence, our research group has demonstrated that increased expression of UCP2 mRNA directly correlates with resistance to gemcitabine treatment, in a panel of pancreatic adenocarcinoma cell lines, and that the UCP2 gene is induced by gemcitabine, demonstrating that the antioxidant effect of UCP2 plays a critical role in pancreatic cancer cell resistance to standard chemotherapy. We have also shown that UCP2 inhibition has a synergistic antiproliferative effect with gemcitabine in pancreatic adenocarcinoma cell growth[57].

Fisetin, a phytopolyphenol, targets apoptotic and necroptotic cell death in HepG2 cells

https://pubmed.ncbi....h.gov/31634424/

Fisetin (3,7,3',4'-tetrahydroxyflavone), a bioactive dietary flavonoid, intrigued scientists for its anticancer potential against various cancer types. We investigated the fisetin-induced inhibition of growth and survival of human hepatocellular carcinoma. Fisetin decreased cell viability and proliferation of HepG2 cells as revealed from MTT and clonogenicity assays. Cell cycle arrest in the G2/M phase was observed. Annexin V/propidium iodide (PI) staining followed by flow cytometry revealed that fisetin induced both apoptosis and necroptosis in HepG2 cells. Apoptotic cells were significantly increased on fisetin treatment as observed in morphological evaluations and 4',6-diamidino-2-phenylindole and Acridine orange staining. Flow cytometry, fluorescence imaging, and 2', 7'-dichlorofluorescein diacetate analyses showed an increase in reactive oxygen species (ROS) generation on fisetin treatment. Pretreatment with N-acetyl cysteine inhibited ROS production and also rescued mitochondrial membrane potential in HepG2 cells. The underlying mechanisms of apoptosis and necroptosis were determined by analysis of their respective signaling molecules using qRT-PCR and Western blotting. Fisetin showed a marked increase in the expression of TNFα and IKκB with a decrease in NF-κB, pNF-κB and pIKκB expression. Fisetin reduced the expression of Bcl2, and elevated levels of Bax, caspase-3, and PARP and thus induced apoptosis in HepG2 cells. zVAD suppressed the fisetin-induced expression of caspase-8, RIPK1, RIPK3, and MLKL as opposed to fisetin treatment. Nec-1 + fisetin could not completely block necroptosis, which warrants further investigation. Taken together, our findings demonstrate that the fisetin exhibited anti-proliferative effects on HepG2 cells through apoptosis and necroptosis via multiple signaling pathways. Fiestin has potential as a therapeutic agent against hepatocellular carcinoma.

Fisetin induces apoptosis in human nonsmall lung cancer cells via a mitochondria-mediated pathway

:text=Fisetin%20induced%20both%20the%20production%20of%20intracellular%20reactive,of%20sub-G1%20phase%20cells%2C%20and%20mitochondrial%20membrane%20depolarization.' class='bbc_url' title='External link' rel='nofollow external'>https://pubmed.ncbi....depolarization.

The present study investigated the apoptotic effects of fisetin, a phenolic compound, against the human nonsmall cell lung cancer cell line, NCI-H460. Fisetin showed dose-dependent cytotoxic activity against NCI-H460 cells, with 50% inhibition of cell viability occurring at a concentration of 75 μg/mL. Fisetin induced both the production of intracellular reactive oxygen species and apoptosis, as evidenced by apoptotic body formation, DNA fragmentation, an increase in the number of sub-G1 phase cells, and mitochondrial membrane depolarization. Moreover, fisetin significantly modulated the expression of apoptosis-associated proteins, resulting in reduced expression of B cell lymphoma-2, increased expression of Bcl-2-associated X protein, and activation of caspase-9 and caspase-3. In addition, pretreatment with a caspase inhibitor blocked fisetin-induced cell death.

Edited by Kelvin, 05 January 2023 - 10:55 PM.

[Kelvin]

Based on these links it looks like UCP2 is over expressed in cancer cells because without UCP2 over expression their heavy use of glycolysis would create higher than normal reactive oxidative species.

This extra oxidation would otherwise cause cancer cells to age and die faster than healthy tissues.

However, the oxidation suppression properties of UCP2 give cancer cells (and stem cells) the quality of cellular immortality.

This is consistent with my hypothesis that suppressing UCP2 with C60 could set off oxidative pressures within cancer cells that would lead to cellular damage or death.

According to the links on Fisetin research Fisetin’s anti cancer properties apparently work by triggering oxidative reactions in cancer cells lines, but those reactions can be neutralized to save the cancer from apoptosis by using an antioxidant like N acetyl carnitine or a caspase inhibitor.

However it is not clear if Fisetin inhibits UCP2 in order to set off oxidative stress, or if it’s pro-oxidant effects use a different mechanism.

To modify the protocol for a more anti-cancer effect all one would have to do is drop glutathione and any other antioxidants on the C60 + fusion day (because anti-oxidants would relieve cancer cells of oxidative stress despite C60).

Then increase the Fisetin dosage on the fission days, perhaps to around 1 or 2 grams of Fisetin to provide extra oxidative stress on any cancer cells that survived the pro-oxidant effect on the C60 day.

Hopefully, the C60 exposure and resulting oxidation processes would leave any surviving cancer cells more vulnerable than normal to a large dose of Fisetin.

This protocol could be repeated every week or twice a week for a month as a cancer preventative protocol.

The paper on UCP2 and pancreatic cell lines did suggest that prolonged UCP2 inhibition (such as in genetically engineered UCP2-null mice) could initiate a tumor, presumably by making healthy cells more susceptible to oxidative damage and free radicals.

But I don’t think this would be a risk with the protocol because the action of C60 in human mitochondria is short lived.

It does suggest taking C60 daily may not be a good idea because of the risk constant UCP2 inhibition of healthy cells would leave them vulnerable to oxidative damage that could lead to tumorogenesis.

Edited by Kelvin, 05 January 2023 - 11:47 PM.

[Kelvin]

I searched again to see if Fisetin inhibits UCP2 but didn’t find any papers that study a connection.

My suspicion is that it does not trigger oxidative stress in cancer cells via UCP2 inhibition because if Fisetin did then it would also trigger stem cell proliferation and be noted for similar health benefits that C60 users (who are taking just C60 and not the Turnbuckle protocol which includes fusion) have experienced.

I suspect the oxidative stress Fisetin inflicts on cancer cells is being induced through some other mechanism.

Edited by Kelvin, 06 January 2023 - 12:46 AM.

[Kelvin]

C60 fullerene and its nanocomplexes with anticancer drugs modulate circulating phagocyte functions and dramatically increase ROS generation in transformed monocytes

https://cancer-nano....2645-017-0034-0

Background

C60 fullerene-based nanoformulations are proposed to have a direct toxic effect on tumor cells. Previous investigations demonstrated that C60 fullerene used alone or being conjugated with chemotherapeutic agents possesses a potent anticancer activity. The main aim of this study was to investigate the effect of C60 fullerene and its nanocomplexes with anticancer drugs on human phagocyte metabolic profile in vitro.

Methods

Analysis of the metabolic profile of phagocytes exposed to C60 fullerene in vitro revealed augmented phagocytic activity and down-regulated reactive nitrogen species generation in these cells. Additionally, cytofluorimetric analysis showed that C60 fullerene can exert direct cytotoxic effect on normal and transformed phagocytes through the vigorous induction of intracellular reactive oxygen species generation.

Results

Cytotoxic action as well as the pro-oxidant effect of C60 fullerene was more pronounced toward malignant phagocytes. At the same time, C60 fullerenes have the ability to down-regulate the pro-oxidant effect of cisplatin on normal cells. These results indicate that C60 fullerenes may influence phagocyte metabolism and have both pro-oxidant and antioxidant properties.

Conclusions

The antineoplastic effect of C60 fullerene has been observed by direct toxic effect on tumor cells, as well as through the modulation of the functions of effector cells of antitumor immunity.

Background

Nanocarbon materials attract growing interest as a platform for drug development, including anticancer preparations (Dellinger et al. 2013; Yang et al. 2014; Chen et al. 2015). Fullerene–biomolecule conjugates exhibit a very high antineoplastic efficiency. For example, the chemical attachment of anticancer preparations such as Paclitaxel and Doxorubicin (Dox) to C60 fullerene results in an improvement of these drugs’ pharmacokinetics and increases their therapeutic efficacy (Liu et al. 2011; Magoulas et al. 2015; Prylutska et al. 2015). Previous investigations from our research group revealed that pristine C60 fullerene possesses a potent anticancer activity per se (Prylutska et al. 2011a, b; Lynchak et al. 2017). In addition, we have demonstrated that conjugation of Dox and Cisplatin (Cis) with the C60 fullerene led to significant increase in its toxicity toward various human tumor cell lines in vitro and greatly enhances the inhibitory effect of these drugs on the growth of Lewis lung carcinoma in vivo (Prylutska et al. 2015a, b, 2017 Panchuk et al. 2015). It is well documented that the underlying mechanism of antineoplastic effect of C60 fullerene-based nanoformulations is related with the combination of the direct effect on tumor cells (modulation of oxidative stress, apoptosis, necrosis, and autophagy), their effect on tumor microenvironment (reduction of the blood supply to tumor tissues), and activation of the host immune system (Dellinger et al. 2013; Harhaji et al. 2007; Shi and Li 2012).

Edited by Kelvin, 06 January 2023 - 01:34 AM.

[ambivalent]

I am not sure if I am misremebering this but does anyone recall if Moussa and Baati used old c60 oil - from a previous study? They cited their preparation method, but would still perhaps have referenced the previous study methid, if so. Save Turnbuckle, Anthony might be the only one who remembers.

 

.

[joesixpack]

Here is the Baati study for everyone's reference.

 

https://c60-france.c...n-Longevite.pdf

[Empiricus]

As I mentioned above, I suspect some of the c60 I was taking may not have been of the highest potency or quality. Following is my review of some brands of c60 available on Amazon and brief descriptions of some side effects that followed.

First a disclaimer. My inclination to attribute the protocol’s main side effects to the c60 may not be correct. For example, after my first three rounds I reduced the Day 2&3 niacinamide dose and replaced most of it with apigenin. Possibly there’s a toxic interaction caused by niacinamide and residual c60 (which i discussed at s i ome length in earlier posts).

- June, stem cell rounds 1-3 (c60 brand A): Side effects were unprecedented clumsiness causing toe injury, nightmares, not needing as much sleep, some memory issues, much improved complexion, general health rebound in following weeks.

- Oct, stem cell round 4 (c60 brand B): Side effects were nightmare, not needing as much sleep, and two nights of dizziness. At Turnbuckle’s recommendation I followed up with mito protocol for two weeks. My overall health and strength improved a lot. Immunity seemed better too.

- Dec, stem cell round 5-6 (C60 brands D&C): Needed to sleep longer, complexion poor, feel run down.

- Dec, took TruMe test of bio age.

- Dec, stem cell round 7-8 (c60 brand D): Need to sleep longer, complexion poor. I feel as if the last 4 cycles have been useless or counter-productive.

A - PureC60OliveOil 99.95%
B - SES 99.99% olive
C - AvaNaturals 99.9% carbon-60 in olive oil
D - Suspended Solutions olive 99.9%

I’m not prepared to recommend any of the above brands. I can’t be sure A wasn’t responsible for the causing the next-morning extreme clumsiness that resulted in an badly injured toe; B had unpleasant side effects, but when followed by mito protocol delivered tangible health improvements; C&D didn’t feel like they were doing anything at all except increase my need for sleep.

If i were to take one of the above again, I’d choose B and cut the dose. I might be willing to give C a second try. I’m done with A and D.

I missed something important in my review. The last two brands say they contain 99.9% c60 (I just edited the above post to show this). When I ordered them, I was so focused on the customer reviews and method of preparation— both companies claimed to brew the c60 in complete darkness—that i missed that neither used even 99.95% purity!

Although I am not sure if SES is the source these two companies use for their 99.9% c60, when you go the SES website, SES tells you their 99.9% c60 powder isn’t prepared the same way as their higher purity 99.95% and 99.99% c60.

Anyway, I think I may have the answer to why these brands left me feeling tired and somewhat unwell: too many impurities. At the time I didn’t realize 99.9% c60 was available, and must have assumed the manufacturer had rounded down, as their descriptions of their processing with regards to avoiding light contamination —a really big concern of mine—sounded so meticulous.

Edited by Empiricus, 07 January 2023 - 06:49 AM.

[Blu]

An interesting meta-analysis on antidepressants and neural stem cells proliferation.

 

https://stemcellsjou...02/sctm.19-0020

[Kelvin]

I’ve been on my low carb diet again to lose extra pounds I gained over Christmas and New Year’s.

Losing weight has been very easy compared to before when I hadn’t tried C60.

I can enter ketosis in less than 24 hours whereas it used to take 2 to 3 days.

I am also triggering ketosis without cutting as many carbs as recommended by people like Dr Atkins or Gary Taubs.

I used to need to reduce my carb intake to under 40 grams a day but now I am losing weight while still eating 100 grams of carbs.

Edited by Kelvin, 11 January 2023 - 03:39 AM.

[ambivalent]

I can enter ketosis in less than 24 hours whereas it used to take 2 to 3 days.

 

 

That's a fascinating result, and would be great to prove several times over (both cases). Any thoughts as to why? 

 

When I used to fast frequently, I played around with c60, before during and after fasts but couldn't recall anything noteworthy - there may have been an observation relating to c60 prior to fasting though possibly an invented memory. I will have to trawl old posts.       

[Answers]

I have read that stem cells can help brain injury and i'm wondering if c60 would be a good candidate for this?

I see discussion of antidepressants also which caused me permanent anhedonia, cognitive problems, dysautonomia and depersonalization. Could boosting stem cells with this substance be helpful for these problems?

[Gern]

I have read that stem cells can help brain injury and i'm wondering if c60 would be a good candidate for this?

Since one study found up to a 90% extension of lifespan, but the results were not replicated in subsequent ones, I’m with Turnbuckle in thinking there have to be other factors in play besides just c60 on its own. Anecdotal evidence seems to bear this out as the effectiveness of c60 appears to diminish over time. If Trunbucklrs theory is correct, stearin acid to promote fusion is a critical component of his protocol, and I don’t believe it crosses the blood brain barrier. Keep in mind I’m a layman and I would have to defer to someone more learned for a more detailed answer.

[Blu]

There are fusion promoters that cross the blood brain barrier.

In this thread there has been discussion about sulforaphane and dihydromyricetin, in that regard.

[Empiricus]

I have read that stem cells can help brain injury and i'm wondering if c60 would be a good candidate for this?

I see discussion of antidepressants also which caused me permanent anhedonia, cognitive problems, dysautonomia and depersonalization. Could boosting stem cells with this substance be helpful for these problems?

 

As it happens, I got a mild concussion about 10 days ago and have been dealing with nausea, headaches, weakness, and some memory issues.    I did a few days of Turnbuckle's mito protocol (alternating niacinamide+ribose+senolytics with dihydromiricetn.  PQQ & AKG every day).  I also took a lot of taurine.  

 

Intermittent fasting and keto diet seemed to be helping the most.  They make a huge difference.  

 

Yesterday, after fasting for 40 hours, I began a round of the stem cell protocol.  I only used dihydromiricetin (DHM) for fusion as it crosses the bbb, and probably the fasting strongly augmented this fusion process. I took the first brew yesterday evening.  I included some astragalus, as I hadn't used it with a stem cell protocol before, and I believe this was my 10th or 11th cycle.   Digesting the brew felt mentally taxing, but the headache that had disappeared by the 30th hour of my fast hasn't returned. The explanation could be that I only had one meal after starting the protocol and then resumed fasting. Anyway, so far so good.  Tonight I take the fission stuff.    

 

There's some evidence DHM, AKG, niacinamide, lysine can be helpful for brain traumas of various kinds, at least in rodents, so the protocol contains various things that would appear to be beneficial. PPQ, a component of the mito protocol, has been suggested as helpful for concussions. On forums, several have reported good results from intermittent fasting and/or keto.

 

Digesting the fusion stuff did feel like doing some really heavy mental work, and left me feeling mentally exhausted, so it's perhaps not a good idea to start this protocol too soon after a head injury.  

Edited by Empiricus, 26 January 2023 - 06:22 PM.

[ambivalent]

Empiricus, I hope you recover well. The c60oo may have helped:

 

https://news.rice.ed...-flow-in-brain/

 

I recall niner mentioning a decade or so ago, that he had given c60oo to his mother in law after a stroke and she made a full recovery. I don't recall the administration window but I wouldn't assume it was just a handful of hours.

Edited by ambivalent, 26 January 2023 - 06:30 PM.

[eighthman]

The way forward in longevity seems clear.  With regard to cells:

 

Kill 'em  (senolytics)

 

Fix 'em  (autophagy)

 

Replace 'em ( stem cells, as here)

 

Steps done in neat, separate cycles may be critical.   #1 and 2 could contradict each other.

[Answers]

Thanks for the responses- I'll try not to sidetrack this thread too much.

I am relatively young (28) and after taking several medications and things my brain is not in good health. I feel that I may need to call upon some stem cells now to try and rebuild my brain after suspected damage and, hopefully, reverse any epigenetic changes caused by antidepressants.

Could someone post the stem cell protocol please? As I have a hard time finding it.

Edited by Answers, 27 January 2023 - 02:29 PM.

[Answers]

There's some evidence DHM, AKG, niacinamide, lysine can be helpful for brain traumas of various kinds, at least in rodents, so the protocol contains various things that would appear to be beneficial. PPQ, a component of the mito protocol, has been suggested as helpful for concussions. On forums, several have reported good results from intermittent fasting and/or keto.

Digesting the fusion stuff did feel like doing some really heavy mental work, and left me feeling mentally exhausted, so it's perhaps not a good idea to start this protocol too soon after a head injury.

Thank you. I did try lots of PQQ and niacinamide to reverse my suspected damage. I also took AKG but I consumed all these substances seperately

Edited by Answers, 27 January 2023 - 02:35 PM.

[ambivalent]

Also, TUDCA, which is or was if I recall, part of the protocol:

 

"Tauroursodeoxycholic acid increases neural stem cell pool and neuronal conversion by regulating mitochondria-cell cycle retrograde signaling"

 

https://www.ncbi.nlm...les/PMC4613652/

 

Unfortunately, Turnbuckle, removed the protocols from his profile - so perhaps someone could be custodian and add them? Or perhaps create a standalone thread for them. It would be a task, but it could be worthwhile creating a thread which narrates the protocol changes from the start, providing reasons for altering them and so forth. 

 

Additionally, Answers, fasting might be considered an option - Mark Mattson gave a TED a few years ago on Fasting and the Brain, though I am sure there are more updated talks. 

Edited by ambivalent, 27 January 2023 - 05:18 PM.

[Heisok]

This is an extended update to post 1711. The updated protocol can be found near the end.

 

 

 

THE DE-AGING HYPOTHESIS

 

The human body is a work in progress. Some 300 billion cells are lost every day and must be replaced by stem cells (SCs). Given that viable SC numbers decline dramatically throughout life even while cellular replacement needs increase, the endogenous proliferation of stem cells should provide the most direct and comprehensive approach to restoring cellular maintenance and extending lifespan.

 

Each of the 200 different cell types in the human body has the same genetic code, but are distinguished by another code that resides above the genetic code and determines which genes are turned on or off for each cell type. This is the epigenetic code. Most of the epigenetic code is in the form of methylation of the DNA or the histones that carry them. 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. It will also reduce the epigenetic age, which can be easily measured in a lab. Tests are available to the public, with one offered by Trumelabs.com presently the cheapest.

 

Cells are unaware of their epigenetic damage, but have a clock to determine when they are old and thus sufficiently damaged to require replacement. This is the telomeric clock. When it runs out (the Hayflick limit) cells commit suicide (apoptosis). Hopefully there are sufficient SCs to supply replacements. But with age, there aren’t, as SC niches become depleted.

 

So how to refill SC niches?

 

It’s known that SCs are quiescent due to high expression of uncoupling protein 2 (UCP2), which allow protons to dissipate through the inner mitochondrial membrane without producing ATP. It is also known that mitochondrial fusion will bias SC division to proliferation. Thus by simultaneously supplying a fusion supplement and a blocker for UCP2, quiescent SCs should awaken and begin proliferating, subsequently replacing senescent cells as needed.

 

A proposed method for reversing age can be divided into three parts:

1. Filling SC niches: Blocking proton leakage through UCP2 pores with C60* activates SCs while promoting mito fusion directs them to proliferation.
2. Replacement of senescent cells: Mitochondrial fission/apoptosis using senolytics removes senescent cells, which are replaced by cells derived from proliferated SCs of part A, at the direction of natural paracrine signaling.
3. Maximizing epigenetic age reversal: Supplying a demethylase promoter during parts A and/or B further reduces aberrant methylation and epigenetic age.

*The fullerene C60 dissolved in oil is proposed herein as a UCP2 blocker. The use of C60 to extend rodent lifespans was the subject of three papers. The first (in 2012, PMID 22498298) showed that feeding rats C60 in oil increased rat lifespans by 90% over controls, and attributed the increase to the antioxidant properties of C60. The next two (in 2021, PMID 33123847 and PMID 33849306) attempted to replicate this result in mice, but found no increase in lifespan. The discrepancy can be resolved by postulating a different MOA for C60 — UCP2 blocking rather than ROS quenching — and a different feeding regime for the three experiments. Although none of the papers precisely described how their test animals were fed, it is likely the research group of the first paper fasted their rats overnight (as they did in a previous C60 toxicology study), while research groups of the next two papers likely did not, as there was no suggestion to do so in the first paper. Rodents have a metabolic rate about six times that of humans, so a state of fasting-induced mitochondrial fusion would have been easily achieved in the first study, but no fusion would have occurred in next two.

 

 

SUPPLEMENTS FOR REDUCING EPIGENETIC AGE

 

 

For fusion, use one or more of the following:

 

Preferred: Dihydromyricetin (DHM): This can penetrate the BBB, but has terrible taste. It should be stirred into water or juice with a high speed blender. It can also be baked into brownies, but ruins the taste.

 

Stearic acid triglyceride (food grade stearic acid): This has low availability unless prepared properly (baked into a brownie, for instance) and taken 3 hours before other supplements due to its slow digestion.

 

Mito fusion brownies

 

Betty Crocker Fudge brownie mix* (519 g)

2 eggs

4-5 tbsp water

120 g food grade stearic acid

 

*Or any mix that requires ½ cup oil, but don’t use the oil. Diet versions don’t work well. 

Mix with power mixer and bake as directed on the box. Cut into 16 pieces, dust with flour to avoid sticking, and freeze. Dosage: 1 piece three hours before SC treatment.

 

Stearic acid monoglyceride (aka glycerol monostearate, GMS): This is a much more soluble and bioavailable form of stearic acid, if stirred into water of juice with a high speed blender. Both types of stearic acid have poor penetration of the blood brain barrier (BBB).

 

For fission: Nicotinamide (NAM). This may optionally be used with a like amount of ribose. Niacin may be used, but most will find the flush objectional. NR might be used, but it is essentially NAM + ribose that requires a time delay to be broken down. Apigenin is yet another possibility, but I prefer nicotinamide.

 

For UCP2 blocking: C60 dissolved in a biocompatible oil.

 

For SC nutrition: Lysine and methionine.

 

For promoting demethylase: AKG or source thereof.

 

 

 

UPDATED AGE REVERSAL PROTOCOL

(with DHM and GMS)

  

 

Day 1: Mito fusion/C60 —

1. C60 — one teaspoon if in olive oil, 1.5-2 teaspoons if in MCT oil

2. Sunflower lecithin — 2-4 grams

3. Dihydromyricetin — 2-6 grams

4. GMS — 1-2 grams

5. Glutathione (reduced or liposomal) — .5-1 grams

6. AKG (alpha ketoglutarate) — 1 gram

7. AAKG (arginine alpha ketoglutarate)  — 2-3 grams

8. SAM-e — 200 mg

9. Lysine — 2-6 grams

10. Methionine — 1-3 grams

 

Astragalus root powder, every 10th treatment — 500 mg

 

Day 2: Mito fission, may be repeated for 2 days —

1. Nicotinamide — 1 gram

2. Ribose — 1-2 grams

3. AAKG  — 2-3 grams (AKG can also be used)

4. Lysine — 2-6 grams

5. Methionine — 1-2 grams

6. Fisetin — 100-200 mg

 

You may skip this fission step for the first month. 

 

Notes:

1. I generally add the C60 stack to water spiked with an orange/tangerine “water enhancer.” I take the fission stack as pills.

2. More lysine and methionine may be taken a few hours later.

3. Threonine (not listed) is optional. I have used 5-10g on occasion, so I can't say with certainty if it helps.

4. The solubility of C60 in MCT oil is less than in olive oil, so I use more with MCT oil.

5. I've dispensed with sulforaphane as a fusion agent, but it might be helpful for some people. 

6. Present schedule varies, but generally 4-5 times a month.

7. Sunflower lecithin helps the bioavailability of DHM and GMS and supplies a source of choline that find useful. Sunflower lecithin powder seems considerably superior to soy lecithin for this purpose.

8. Lysine and methionine provide SC nutrition.

9. Astragalus root powder provides a telomerase promoter, but using too much will ruin the epigenetic benefits, as it will lengthen the telomeres of TACs and somatic cells, and block their replacement.

10. 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.

 

Caveats:

1. This is a work in progress.

2. It is intended as a geriatric treatment for age reversal.

3. One should avoid alcohol during this treatment. Do not add other random supplements. Telomerase promoters like resveratrol should definitely be avoided.

4. A link to the latest protocol can always be found on my profile page.

5. All amounts are approximate and based on a 180 pound individual.

  

Results:

Best epigenetic age result to date was 28 years below chronological. Baseline result in 2018 was half a year above chronological. Substantial improvements in appearance and general health, consistent with the drop in epigenetic age.

 

[Empiricus]

Thank you. I did try lots of PQQ and niacinamide to reverse my suspected damage. I also took AKG but I consumed all these substances seperately

 

You're welcome, Answers.  Before he left us, Turnbuckle advised several of us to alternate the stem cell protocol (reposted above by Heisok) with the mitochondria dynamics protocol (posted here).  The latter protocol includes PQQ.  

 

For the purpose of assisting the brain to recover, for both protocols, the fusion component needs to include dihydromiricetin (DMH).  Turnbuckle recommends the addition of DHM in posts on this page of the mito protocol thread:

I noted a dramatic improvement in mental functioning when I added DHM to my C60/fusion SC proliferation protocol, indicating that DHM was more effective than stearic acid in the brain.

 

I just completed a round of the stem cell protocol.  It went very well. Even though I took a break from intermittent fasting, I am feeling a lot better.  I might do one more cycle of stem cell before starting the mito dynamics protocol which could take around 2 weeks (unlike the stem cell protocol, the duration of the mito protocol is determined by the results of exercise tests).  

Edited by Empiricus, 28 January 2023 - 03:58 PM.

[Dallasboy]

You're welcome, Answers.  Before he left us, Turnbuckle advised several of us to alternate the stem cell protocol (reposted above by Heisok) with the mitochondria dynamics protocol (posted here).  The latter protocol includes PQQ.  

 

 

What would be a good schedule to alternate these two protocols?

[Kelvin]

For anyone with brain trauma I recommend my modification of Turnbuckles protocol that includes 5-HTP.

It is a natural SSRI and helps circulate neural stem cells.

I think it may circulate stem cells to a certain degree in the parts of the brain that do not have them because I noticed a cognitive boost when I included it that I didn’t experience when I used the C60 protocol without 5-HTP.

Just remember to use TMG as your methyl donor instead of SAM-e because it is not recommended to mix 5-HTP with SAM-e.

In general I recommend the protocol for those who have used SSRIs of any kind (both natural and pharmaceutical) because SSRIs promote neurogenesis, initially, but probably deplete areas of the brain that do have stem cells. This is why SSRI users often experience a crash after initially improving on them.

WARNING - I strongly recommend replacing SAM-e with TMG for a methyl donor source regardless of whether one uses 5-HTP with the protocol. The reason I dropped SAM-e and now only use TMG for methyl groups is not because SAM-e doesn't work but because 5-HTP should not be combined with SAM-e and I don't want risk accidentally using SAM-e at the same time I am using 5-HTP with the protocol.

It is easier to avoid mixing them up by never using SAM-e with the C60 cycle, regardless of whether or not I am using 5-HTP.

However, I continue to use SAM-e once month with phosphatidylcholine when I am not on the C60 protocol because of the liver protective effects of SAM-e and phosphatidylcholine.

I have also dropped Nicotinamide Riboside chloride both to save money and because NRcl is too slow acting.

I have dropped d-Ribose as unneccessary.

Instead I now take 1 gram of Nicotinamide for fission on Fission day 1 and then NMN on Day 2 as the only fission and NAD+ promoter.



I am making this change because, although NMN causes fission in most of the body, it seems to cause mito fusion in the brain.

Therefore I will alternate it with the fission and NAD+ promoter, Nicotinamide.

I want to keep NMN as a fission and NAD+ promoter because of its very impressive results on muscles, cardio-vascular system, and other health benefits I have experienced using NMN with both the C60 protocol and the mitochondrial protocol.



Here is my C60 protocol.

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

Also I do not make 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.

I only take it out of the freezer the night before I use it and place it (still wrapped in aluminum) in a dark closet overnight. After I briefly take the 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, otherwise I get nauseus if I try to take the capsules all at once-



***** 0 hour *****

2 to 3 grams Dihydromyricetin (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 (Swanson Vitamins brand)



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

1 gram Liposomal Glutathione
1 gram AKG (Now using Double Wood's AKG instead of Simplesa)
100 mgs Sulforaphane (2 capsules)
500 or 1,000 mgs TMG

--------> 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. Do not combine SAM-e and 5-HTP)


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

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

2 grams Methionine
2 grams Lysine

3 mgs C60 oil capsules (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 capsule on hand whenever doing the C60 cycle.



*** Fission/Senolytics Day 1. The day after taking C60 take senolytics and fission promoters once in the morning, but not for more than 3 days. One could also drop the senolytics if you aren't seeing benefits and just use AAKG, Nicotinamide or NMN for fission promotion.


3 grams AAKG
500 mgs Fisetin (Fission promoter and Senolytic. Can be reduced to 100 mgs if one stops feeling an effect from them)
500 mgs Quercetin (Senolytic)
1 gram Nicotinamide (Fission and NAD+ promoter)
(Optional) 1 gram Curcumin (Senolytic but promotes fusion. Use only occasionally with fission promoters to trigger apoptosis of senescent cells because combining fission promoters with fusion promoters may work against eachother)


*** Fission/Senolytics Day 2.


3 grams AAKG
500 mgs Quercetin (Senolytic)
1 gram NMN (Fission and NAD+ promoter (except in the brain where it promotes fusion). When using NMN do not use it with other fission promoters or with other NAD+ promoters because NMN causes fusion in the brain)

[Kelvin]

I have read that stem cells can help brain injury and i'm wondering if c60 would be a good candidate for this?

I see discussion of antidepressants also which caused me permanent anhedonia, cognitive problems, dysautonomia and depersonalization. Could boosting stem cells with this substance be helpful for these problems?

If you were on SSRIs your neural stem cell pool is likely depleted (who knows what mental disorders have been causing for the general population that has been subscribed SSRIs…)

Try my version of the protocol that uses 5-HTP that I just posted.

Without it Regenerating stem cells in the brain would only work in areas like the hippocampus that already have stem cells, but it would probably have little to no effect on areas like the frontal lobe that do not have stem cells.

Turnbuckle suggested an SSRI like 5-HTP could circulate stem cells across the brain more generally.

In my experience there was a cognitive boost to adding 5htp that I did not get when I tried the protocol without it, which makes me suspect it did (to some extent) get stem cells to areas of the brain that do not have them.

[Kelvin]

Thank you. I did try lots of PQQ and niacinamide to reverse my suspected damage. I also took AKG but I consumed all these substances seperately

You should take them at the same time, although I don’t think the mito protocol has as dramatic an improvement in cognition as the C60 protocol.

I had tried the mito protocol 3 years before I tried C60 and it was only when I tried C60 that I got mental improvements like greatly reduced need for sleep (I can function ok with 5.5 hours and more than 6.5 feels excessive whereas before I wasn’t satisfied with 8 hours).

Adding 5-HTP worked even better than when I tried the C60 protocol without it.

Edited by Kelvin, 31 January 2023 - 12:12 AM.

[Kelvin]

You're welcome, Answers. Before he left us, Turnbuckle advised several of us to alternate the stem cell protocol (reposted above by Heisok) with the mitochondria dynamics protocol (posted here). The latter protocol includes PQQ.

I recently tried a couple rounds of my version of the mito protocol and fusion really knocked the stuffing out of me, like the first time I tried mito fission/fusion 4 years ago.

I think my mitochondria was suboptimal because I had created so many stem cells that good mitochondria became more scarce in my body.

Side effects gradually reduced until the mito protocol had no side effects but I am going to increase my use of them mito cycle to 2 to 3 cycles a month just to make sure I don’t let mito damage buildup again.

Edited by Kelvin, 31 January 2023 - 12:19 AM.

[Answers]

If you were on SSRIs your neural stem cell pool is likely depleted (who knows what mental disorders have been causing for the general population that has been subscribed SSRIs…)

Try my version of the protocol that uses 5-HTP that I just posted.

Without it Regenerating stem cells in the brain would only work in areas like the hippocampus that already have stem cells, but it would probably have little to no effect on areas like the frontal lobe that do not have stem cells.

Turnbuckle suggested an SSRI like 5-HTP could circulate stem cells across the brain more generally.

In my experience there was a cognitive boost to adding 5htp that I did not get when I tried the protocol without it, which makes me suspect it did (to some extent) get stem cells to areas of the brain that do not have them.

I had no idea they depleted stem cells until I came to this thread.

I won't pretend to know how any of this works as I find it pretty complex. But if I did have depleted stem cells, could that be a reason why my brain can't find homeostasis or seem to repair itself? And why the effects of taking antidepressants continue after being off them for years?

Can stem cells reverse potential epigenetic changes?
Taking 5htp makes me nervous as my efforts and everything I've researched has been towards reducing serotonin, as it's linked to autism, PSSD and anhedonia.

The frontal lobe and other brain areas is exactly where I would want stem cells to go

[Kelvin]

I had no idea they depleted stem cells until I came to this thread.

I won't pretend to know how any of this works as I find it pretty complex. But if I did have depleted stem cells, could that be a reason why my brain can't find homeostasis or seem to repair itself? And why the effects of taking antidepressants continue after being off them for years?

Can stem cells reverse potential epigenetic changes?
Taking 5htp makes me nervous as my efforts and everything I've researched has been towards reducing serotonin, as it's linked to autism, PSSD and anhedonia.

The frontal lobe and other brain areas is exactly where I would want stem cells to go

I have often wondered if neural stem cell depletion from SSRIs has anything to do with the spate of school shootings since the late 1990s.

Stem cells are needed for repairing every type of cell in the body.

Yes the protocol will restore epigenetic youthfulness.

If you are nervous about 5-HTP then you can try 3 to 6 cycles of my protocol EXCEPT for the 5-HTP.

Then try 2 cycles with the complete protocol I just reposted.

After what you learned about SSRIs you might be nervous about trying them again. But 5-HTP, like all SSRIs, does stimulate neurogenesis as long as the stem cell pools of the hippocampus, and other areas of the brain that have stem cell reserves, are plentiful.

An SSRI therefore might be the only way to circulate stem cells to parts of the brain like the frontal lobe that have no, or very little, access to stem cells.

Give my protocol first without 5-HTP and then with it a try and then report back your results.

If you have a negative reaction take a break and ask questions.

Edited by Kelvin, 31 January 2023 - 01:54 AM.