2503 replies to this topic

[ambivalent]

Thanks for the reply. The benefit you described, though, was considerable, would you say much of it has been retained? What is your likely strategy from here, fasting and senolytics for months followed up by the stem cell protocol again? 

[Turnbuckle]

Thanks for the reply. The benefit you described, though, was considerable, would you say much of it has been retained? What is your likely strategy from here, fasting and senolytics for months followed up by the stem cell protocol again? 

 

All of it was retained. My present goal is to find a suitably effective senolytic treatment for age regression.

[Nate-2004]

So the goal of deaging requires the refilling of stem cell pools, but also requires the elimination of old cells. Either one by itself is inadequate.

 

This is why I've been doing extended fasting (sometimes while taking dasatinib, fisetin, quercetin and apigenin) in the week leading up to starting your stem cell protocol for about 3 days after I start refeeding again. 

 

 

 

(3) telomerase stimulants are a mistake as non-stem cells are doing most of dividing in many organs and resetting their telomeres adds to epigenetic aging. For example, in the skin these are transit amplifying cells. See this post in "Reversing the Clocks."

 

What about during this protocol while attempting to induce symmetric division of stem cells using C60 and Stearic Acid?  I've been taking vitamin C, NAC and Liposomal Astragalus along with it.

Edited by Nate-2004, 08 December 2018 - 03:05 AM.

[sthira]

This is why I've been doing extended fasting (sometimes while taking dasatinib, fisetin, quercetin and apigenin) in the week leading up to starting your stem cell protocol for about 3 days after I start refeeding again.

So that's cool. I tried this approach last year on a few occasions, that is, combining 5-7 day water-only fasts with D+Q. I didn't notice anything (what would I notice) beyond the negatives and positives thart fasting alone offers. So eventually I figured screw it, maybe these compounds are not even safe, maybe I'm overdoing it, give it a rest with the extra compounds already. As if I can outsmart millions of years of evolution -- the hubris... Then someone mentioned mixing fisetin with oil to assist with better bioavailability and I though hmm. Then I thought maybe this entire activity of attempting to slow aging is a type of mental disease -- obsessively and compulsively seeking to fulfill some story in my mind.

Have you noticed anything (since we're not blood testing), and if not, what would you expect to notice by adding additional compounds to an already powerful intervention?

I get all kinds of crazy upside and downside effects from fasting. But I'm not pretending that I'm slowing the aging process itself; I'm staying healthy as the aging story continues.

[Nate-2004]

I'm the one who mentioned adding MCT oil to improve bioavailability for the 4 types I was taking during the fast, only a tsp as I didn't wanna break the fast at all. After this latest fast, the one where I did the above described regimen, I no longer have a problem with my fasted blood glucose. May have been the synolitics, may have just been the number of extended fasts I've done over the past 6 months. The next one is coming up. I was 106 to 115 for fasted blood glucose every single morning, now I'm at 80 to 90.

Edited by Nate-2004, 08 December 2018 - 05:28 AM.

[sthira]

...my fasted blood glucose...I was 106 to 115 for fasted blood glucose every single morning, now I'm at 80 to 90.

And lowering of blood sugar, I believe, is one of the most important indicators we can actively monitor. Fasting gives it to us for nearly free (until we stop fasting, and levels may slowly rise again according to whatever diet and activity patterns we follow).

When continuous blood glucose monitors come up to speed, and they're more widely available to non-diabetics -- wow. I want one. To know is power, and knowing changes behavior. Even behavior I may think is healthy may not be. Continuously monitored inflammatory markers and continuously monitored stem cell fluctuations would also give us more insight into the biology we can control.

[Nate-2004]

I didn't start measuring my blood sugar in the mornings before eating until a week or so after the fast and after the stem cell protocol which I began a couple of days after I began refeeding. I measured it on a couple of different days. I'm coming up on a month since the fast and honestly I'm afraid to measure it again hahaha, considering the holiday meals I've had.

 

So I plan to do the same exact protocol again a week from now starting with the fast, the "4 horsemen" as we could call it, then the refeed, then the stem cell protocol. I want to just finish off the C60 I made because it's just sitting there going bad otherwise. I suppose I could freeze it afterward though but I like to start fresh each time I do this since I'm using EVOO and not MCT.

Edited by Nate-2004, 08 December 2018 - 06:03 PM.

[QuestforLife]

Why not combine the Stem Cell expansion and Senolytic portion of this protocol to take advantage of the growth inducing effect of apoptosis products?

 

I've discovered this effect whilst researching my Alternative Methods for Lengthening Telomeres:

 

https://www.longecit...meres/?p=864534

 

 

The correlation between apoptosis of irradiated feeder cells and the release of feeder factors that promote conditional immortalization suggests that irradiation induces these factors and/or that the factors are released from dying cells.

 

https://ajp.amjpatho...0594-4/fulltext
 

 

 

Edited by QuestforLife, 11 December 2018 - 10:29 AM.

[lost69]

today i tried the original recipe from the study and it is by far extremely easy and fast to prepare.the taste is amazing, easy to drink and digest

 

i first made an extremely fine powder of Sigma-Aldrich W303518 by a coffee grinder.it is easy to mix and eat 24g by making a very fine powder

 

i put some milk on the powder, mixed a little and microwaved to reach boiling point

 

in a separate bowl i did banana milk shake

 

i added the hot stearic acid/milk to the banana shake and blended some more and drank straight away.

 

i did not use all the milk like in the study (i think they used 500ml in total) but much less maybe about 250ml in total, the stearic acid was not solid even if it was not hot while drinking

i think that boiling in microwave until milk becomes a tranparent liquid and blending it well with this liquid, or remaining concentrated milk, makes it possible to drink it not hot and not solid.i guess as turnbuckle said it bonds to the milk molecules and this way it stays liquid some more when cooling off

 

i don t know if this recipe can work with the commercial stearic acid, the sigma stearic acid is very diiferent in the way it melts and solidifies again

 

Edited by lost69, 16 December 2018 - 05:17 PM.

[jgkyker]

I didn't read through the entire thread. So, forgive me if this idea has already been posted.

 

Rather than buy stearic acid that is ~50%+ palmitic acid, I have decided to go with cocoa butter. It is ~25% palmitic acid and is ~30% stearic acid, tastes better (put it in coffee), melts more easily, and is fairly inexpensive. From what I remember, eating cocoa butter has fewer consequences than other high-fat foods, but I do not have a study to post on that at the moment.

 

Fat Composition of 100g Cocoa Butter

"59.7 g of total saturated fat. Some of these saturated fatty acids are 33.2 g of Stearic Acid (SA), 25.4 g of Palmitic Acid (PAL) and 0.1 g of Myristic Acid (MA). The amount of Caprylic Acid (CA) and Lauric Acid (LAA) are zero or negligible."

 

1 lb of food grade cocoa butter on Amazon is less than $15

 

16 oz ~= 453 grams * ~30% stearic acid content = 135.9 grams stearic acid

 

At a 10 gram target of stearic acid per dose, that is roughly 13 doses. Granted, it isn't pure stearic acid, but neither is that palmitic acid mix. My guess is the cocoa butter is healthier, and this is why I chose this route.

 

The most interesting part of all this discussion reminds me of how incredible chocolate tastes and smells. It is intoxicating. I wonder if the body knows the high level of stearic acid in it and gives us that intoxicating effect to try to get us to promote mitochondria fusion...

 

Also, regarding stearic acid as topical treatment, cocoa butter seems to be a very popular topical treatment for skin. At least, many people on Amazon seem to be very happy with it (NOTE that prior link is probably NOT food grade). I am considering it... but will probably pass for now. I'm not really big into topical treatments at this time.

Edited by jgkyker, 18 December 2018 - 05:28 PM.

[jgkyker]

Any thoughts on this study? It seems to indicate that NMN (a NAD+ precursor) promotes fusion rather than fission.

 

Effect of nicotinamide mononucleotide on brain mitochondrial respiratory deficits in an Alzheimer’s disease-relevant murine model

"Furthermore, we found a shift in dynamics from fission to fusion proteins in the NMN-treated mice."

 

If I'm following, we have research indicating nicotinamide promotes fission:

"Here, we present evidence to show that the effect of nicotinamide is mediated through an increase of the [NAD+]/[NADH] ratio and the activation of SIRT1, an NAD+-dependent deacetylase that plays a role in autophagy flux. The [NAD+]/[NADH] ratio was inversely correlated with the mitochondrial content, and an increase in the ratio by the mobilization of the malate-aspartate shuttle resulted in autophagy activation and mitochondrial transformation from lengthy filaments to short dots."

 

However, we have NMN research indicating a shift to fusion... perhaps, this is why Sinclair saw exercise performance increases in his study:

"Treatment of mice with the NAD + booster nicotinamide mononucleotide (NMN) improves blood flow and increases endurance in elderly mice by promoting SIRT1-dependent increases in capillary density, an effect augmented by exercise or increasing the levels of hydrogen sulfide (H 2S), a DR mimetic and regulator of endothelial NAD + levels. These findings have implications for improving blood flow to organs and tissues, increasing human performance, and reestablishing a virtuous cycle of mobility in the elderly."

 

And this study also by Sinclair:

"Without any obvious toxicity or deleterious effects, NMN suppressed age-associated body weight gain, enhanced energy metabolism, promoted physical activity, improved insulin sensitivity and plasma lipid profile, and ameliorated eye function and other pathophysiologies."

 

Whereas NR (promotor of fission) is indicating exercise loss:

The NAD⁺ precursor nicotinamide riboside decreases exercise performance in rats

"The nicotinamide riboside group showed a tendency towards worse physical performance by 35 % compared to the control group..."

 

So, I'm beginning to wonder if NMN should be cycled against NR? Perhaps something like:

Day 1: Fission day (NR etc.)

Day 2: Fusion day (NMN, Sulforaphane, C60, Stearic Acid)

Edited by jgkyker, 19 December 2018 - 05:42 PM.

[jgkyker]

I may be missing something here, but our body is constantly in a state of mitochondrial fission/fusion. Isolated areas can change state in a matter of seconds. The idea here is to "promote" fusion. That does not eradicate fission. It would just lessen it. With that said, fission will continue to take place, of course. Consequently, why does the scale ever need to be tipped into the fission direction? Why would I need to spend a day promoting fission over fusion? My understanding is the idea is that promoting fission will help the body clean-up defective mitchondria. However, this is still happening even on the fusion promotion days, perhaps less so. Why have a fission promotion day/period at all? The Sinclair studies seem to indicate successive periods of fusion promotion are okay, and so does the C60 study showing the extraordinary life expectancy improvement. Did I miss something? (I probably did.)

 

C60 Study Full-Text with Graphs Showing Enhanced Survival Rate

[Kentavr]

I may be missing something here, but our body is constantly in a state of mitochondrial fission/fusion. Isolated areas can change state in a matter of seconds. The idea here is to "promote" fusion. That does not eradicate fission. It would just lessen it. With that said, fission will continue to take place, of course. Consequently, why does the scale ever need to be tipped into the fission direction? Why would I need to spend a day promoting fission over fusion? My understanding is the idea is that promoting fission will help the body clean-up defective mitchondria. However, this is still happening even on the fusion promotion days, perhaps less so. Why have a fission promotion day/period at all? The Sinclair studies seem to indicate successive periods of fusion promotion are okay, and so does the C60 study showing the extraordinary life expectancy improvement. Did I miss something? (I probably did.)

C60 Study Full-Text with Graphs Showing Enhanced Survival Rate

Formula 1: min(igf-1) --> max(foxo1) --> + own stem cells (stimulate) Very simple!

[jgkyker]

Formula 1: min(igf-1) --> max(foxo1) --> + own stem cells (stimulate) Very simple!

 

Thank you for this. I will research further. I believe what is implied here is that the mice in the C60 study could have lived even longer if they had cycled fission/fusion promotion, and this is why the OP is suggesting stearic acid fusion promotion with C60 in opposition to fission cycle days with N+R.

[QuestforLife]

Thank you for this. I will research further. I believe what is implied here is that the mice in the C60 study could have lived even longer...

No one knows why the rats in the study lived so long. The authors proposed it was due to the reduction in oxidative stress. Apparently the rats also stayed free of cancer, which is unusual.

Turnbuckle believes C60 stimulated their stem cell production and this protocol is an effort to recapture these benefits in humans, which requires stearic acid to sustain the benefits of C60.

[orion22]

Turnbuckle aren t you worired about Glycation  from the d-ribose you take with you re nam

and wodn t this explain why you re test showed some genes of you res where older after the protocol?

Edited by orion22, 20 December 2018 - 05:10 PM.

[Fafner55]

My present thinking on this protocol is summarized below—

 

1. Taking telomerase supplements on a regular basis is a mistake. In fact, it may always be a mistake. Telomere length is a tool by which the body protects itself from cancer and marks old cells for destruction. Lengthening telomeres injudiciously may produce a short-term improvement in health by rescuing senescent cells, but will ultimately allow cells to age beyond their sell-by date.

 

2. Filling stem cell pools produces a noticeable improvement in youthfulness, but overfilling them produces no further apparent improvement. Old cells don’t die just because new cells are available. With a pseudo-geometric progression of stem cells via fusion/C60 treatment, just a few cycles of the stem cell protocol may be all that is necessary. Three will make a difference (though it might take weeks to see it), and there may be no further improvement after a few dozen.

 

3. Eliminating old cells (and not just senescent cells) allows stem cell replacement and restoration of health. Once stem cells are replenished, this is the bottleneck.

 

4. Apoptosis (programmed cell death) is just as necessary to restoration of youth as filling stem cell pools. In ancient medicine, purgatives were used to restore health, and some of these were quite toxic.  Black hellebore, for instance, was the most popular among the Greeks. Today hellebore is known to contain substances that stimulate apoptosis via the caspase mechanism.

 

5. Caspases are suicide enzymes present in precursor form in all cells. These procaspases must be activated to create a “caspase cascade,” which is like a nuclear chain reaction inside the cell. Once started, it accelerates and cannot be stopped. Since it is programed, apoptosis is orderly and much less toxic to the body than necrosis.

 

6. Adults are said to lose more than 50 billion cells to apoptosis every day, which is about 0.1% of the total, though most of this is limited to a few organs. If dead cells are replaced by somatic cells, this continues the aging process as somatic cells become epigenetically older every time they divide. With depleted stem cell pools, somatic cell replacements predominate and aging accelerates.

 

7. The goal is then to increase the apoptosis of somatic cells and to bias their replacement by stem cells from full stem cell pools (#3 above) rather than by somatic cells.

 

 

And here I’m a bit up in the air. Could one maximize #7 by stimulating stem cells (ie, C60 without fusion) and stimulating apoptosis at the same time?

 

Increased mitochondrial mass leads to greater autophagy.

"Mitochondrial levels determine variability in cell death by modulating apoptotic gene expression" (2018) https://www.ncbi.nlm...les/PMC5785974/

 

This result might imply that PQQ should be part of a senolytic treatment, and it raises questions about what states mitochondria should be driven (fission or fusion; high or low ROS) to enhance clearance of senescent cells.

 

What are this community's thoughts on the subject of manipulating mitochondrial dynamics for more effective invocation of apoptosis of aged and senescent cells?

[Turnbuckle]

Turnbuckle aren t you worired about Glycation  from the d-ribose you take with you re nam

and wodn t this explain why you re test showed some genes of you res where older after the protocol?

 

No, I'm not worried about ribose and glycation. First, there is no ribose in this protocol. Second, ribose does not impact epigenetic age that I know of. Epigenetic age will increase with telomerase stimulants as such stimulants allow somatic cells to divide more times, and epigenetic age increases with the number of replications. Each replication adds more epimutations, and epimutations occur far more frequently than mutations of the underlying DNA.

Edited by Turnbuckle, 27 December 2018 - 07:51 PM.

[Nate-2004]

No, I'm not worried about ribose and glycation. First, there is no ribose in this protocol. Second, ribose does not impact epigenetic age that I know of. Epigenetic age will increase with telomerase stimulants as such stimulants allow somatic cells to divide more times, and epigenetic age increases with the number of replications. Each replication adds more epimutations, and epimutations occur far more frequently than mutations of the underlying DNA.

 

Does this matter though? Wouldn't these also increase telomerase in stem cells?

[Turnbuckle]

Does this matter though? Wouldn't these also increase telomerase in stem cells?

 

It does matter as in many organs stem cells are only the backup to transit amplifying cells, which do most of the dividing. See this post.

[jgkyker]

Increased mitochondrial mass leads to greater autophagy.

"Mitochondrial levels determine variability in cell death by modulating apoptotic gene expression" (2018) https://www.ncbi.nlm...les/PMC5785974/

 

This result might imply that PQQ should be part of a senolytic treatment, and it raises questions about what states mitochondria should be driven (fission or fusion; high or low ROS) to enhance clearance of senescent cells.

 

What are this community's thoughts on the subject of manipulating mitochondrial dynamics for more effective invocation of apoptosis of aged and senescent cells?

 

Here are my thoughts in the mitochondria dynamics thread. In short, it looks like they go hand-in-hand.

[QuestforLife]

It does matter as in many organs stem cells are only the backup to transit amplifying cells, which do most of the dividing. See this post.

 

There is evidence that longer telomeres lead to (less frequent cell replacement and therefore) more advance epigenetic age (as measured by the Horvath clock of methylation changes). This evidence is mainly from in vitro studies (done by Horvath and not very well in my opinion), and also from epidemiological studies (again, these are questionable because of many confounding factors). But still, I tend to agree that being born with naturally longer telomeres will tend to mean that your Horvath clock will be older.

 

But it is a leap from there to say that telomerase activating molecules will necessarily advance the epigenetic clock. This is because, as Nate says, stem cells will also have their telomeres elongated. We also know that progenitor cells have active telomerase and therefore it may be easier for telomerase activators to extend their telomeres than those of somatic cells. Of course there are many more somatic cells, so the net result is difficult to predict. In any case my point is the work has not been done, so we can't say for sure what will happen either way.

 

One way to resolve this issue would be for those Longecity members who have used telomerase activators long term to take an epigenetic age test to see if their Horvath clock has been accelerated (and if this has in turn lead to a deterioration in health).

 

One anecdotal account - Ed Park, who has used telomerase activators for 10 years, had his epigenetic age measured last year, and he came out as -7 years (so significantly younger epigenetically than his chronological age, not older). You can read about it on his blog.

Edited by QuestforLife, 10 January 2019 - 10:13 AM.

[Kentavr]

One anecdotal account - Ed Park, who has used telomerase activators for 10 years, had his epigenetic age measured last year, and he came out as -7 years (so significantly younger epigenetically than his chronological age, not older). You can read about it on his blog.

Please give a link to his blog.

[QuestforLife]

Please give a link to his blog.

 

https://www.recharge...s-your-dna-age/

[lost69]

too bad he did not take 2 tests, one before intervention or at least few years back from now

[NeilR]

Turnbuckle, thank you very much for all of your work. Do you mind giving me your advice? I’m 31 years old but doing anti-aging stuff and would like to keep my stem cells intact and am very interested in your protocols here.

I assume you wouldn’t recommend your c60 protocol to me due to my age, what would you recommend though?

I’m currently taking Niacin (2 g), D-Ribose (5 g), Icaarin (100 mg), Taurine (5 g) 4 days a week. I’m planning on adding Resveratrol and stearic acid and am considering Metformin, ALA, TUDCAvand Apigenin.

You mentioned that c60 isn’t good for people under 60, so is there a protocol that would work on my stem cells without it?

[Rocket]

I think Niacin and Resveratrol are contraindicated. One will block Sirt2. One will activate it. 

[Turnbuckle]

Turnbuckle, thank you very much for all of your work. Do you mind giving me your advice? I’m 31 years old but doing anti-aging stuff and would like to keep my stem cells intact and am very interested in your protocols here.

I assume you wouldn’t recommend your c60 protocol to me due to my age, what would you recommend though?

I’m currently taking Niacin (2 g), D-Ribose (5 g), Icaarin (100 mg), Taurine (5 g) 4 days a week. I’m planning on adding Resveratrol and stearic acid and am considering Metformin, ALA, TUDCAvand Apigenin.

You mentioned that c60 isn’t good for people under 60, so is there a protocol that would work on my stem cells without it?

 

I have stated several times that this geriatric protocol is experimental. It is for refilling your stem cell niches and using that to reverse epigenetic age, so unless you have reason to believe that your stem cells are depleted, or that stem cell numbers need to be enhanced for a specific purpose (injury or surgery or the like), or that your epigenetic age is increasing too rapidly, using this protocol is not recommended.  After building up my own stem cell pools, I have continued stimulating stem cells at appropriate times without C60. I use 10 grams each of threonine and leucine after senyloic treatment, for instance, or prior to exercise. Pluripotent stem cells have a critical need for threonine and muscle satellite cells are triggered by leucine.*

 

As for your N+R stack, this is off topic on this thread. My N+R protocol was directed to eliminating dysfunctional mitochondria, but someone 30 years old is unlikely to have any. I suggest you get your genetics tested, and if you have one or more APOE4 genes, then there might be a reason to take nicotinamide.

 

*The effect of leucine on muscle gain has been known for some time. It operates by increasing satellite cells, which then mature into muscle cells. The discovery that mice and human pluripotent cells are so dependent on threonine is very new. Even the presence of pluripotent cells in the adult is new and not totally accepted. That said, threonine supplementation is not recommended for daily use, but only as part of this protocol. Don't feed your stem cells when they don't need feeding. 

 

 

Edited by Turnbuckle, 14 January 2019 - 03:20 PM.

[Rocket]

I think Niacin and Resveratrol are contraindicated. One will block Sirt2. One will activate it. 

 

I made a mistake 2 instead of 1. Same issue though. One blocks and one activates.

[Nate-2004]

https://www.recharge...s-your-dna-age/

 

That test doesn't measure telomere length and increasing length is known to speed up methylation age. Not sure about this one.