2503 replies to this topic

[QuestforLife]

Fibrotic problems after senolytic treatments have been found before, I read a study where it happened in the endothelial lining of the cardiovascular system. It was fixed by using a ROCK inhibitor.

Presumably this would only be a problem with Turnbuckle's protocol if you didn't have enough stem cell ready replacements.

[Andey]

Fibrotic problems after senolytic treatments have been found before, I read a study where it happened in the endothelial lining of the cardiovascular system. It was fixed by using a ROCK inhibitor.

Presumably this would only be a problem with Turnbuckle's protocol if you didn't have enough stem cell ready replacements.

 

  I am not sure, they had this problem with young mice where stem pool is presumably full. 

 

TBH it could be that its not applicable to humans too, it could be mice specific problem as they dont have to maintain tissue integrity for long so evolution hasn't optimized it.

Edited by Andey, 20 May 2020 - 08:04 AM.

[Turnbuckle]

 

One could argue that we dont really want too effective senolitic treatment unless this is resolved. 

 

 

I agree. Losing a limb could be considered the maximum end point of cell removal, and while a few animals can replace lost limbs, humans cannot. Thus there must be some maximum cell loss that can be repaired via stem cells without creating chaos. Less than that and you maintain order; more than that and you get various types of scarring.

Edited by Turnbuckle, 20 May 2020 - 09:02 AM.

[kurt9]

Fibrotic problems after senolytic treatments have been found before, I read a study where it happened in the endothelial lining of the cardiovascular system. It was fixed by using a ROCK inhibitor.

Presumably this would only be a problem with Turnbuckle's protocol if you didn't have enough stem cell ready replacements.

 

This reinforces my belief that senolytics should not be used outside of a larger process, such as either the Mitochondrial fission/fusion or this C60 process. Senescent cell clearance is clearly a part of a larger process, and that larger process has to be activated for proper results.

[QuestforLife]

This reinforces my belief that senolytics should not be used outside of a larger process, such as either the Mitochondrial fission/fusion or this C60 process. Senescent cell clearance is clearly a part of a larger process, and that larger process has to be activated for proper results.

And as others have said an environment where excessive inflammatory/fibrotic signalling is suppressed is also required as part of this, probably related to the other topics you mention like mitochondrial state and stem cell availability. Generally in my experience tgf-b is the master switch, which if turned up too much or for too long, results in fibrosis.

[Turnbuckle]

Update on my epigenetic age.

It continues to tick backwards. The last 3 tests from TruMe --

 

Date      Epigenetic age - Chronological age, years

02/2018.... +0.5 (Baseline before treatments)

11/2019.... -13.0

02/2020.... -14.2

04/2020.... -14.5

 

These treatments presently comprise a stem cell enhancement step once every 10-14 days, and a senolytic + stem cell nutrition step 24 hours later, with another the next day. I've recently added a telomerase shortening agent to the senolytic/nutrition steps, and we'll see how that goes with the next results. This agent is cinnamaldehyde (see the paper below), which is the major component of cinnamon oil. I previously found that telomerase enhancers increased epigenetic age, so its logical to try going the other way. 

The cinnamaldehyde and eugenol could be considered as the potential chemotherapeutic agents for cancer prevention via telomere shortening or telomerase blockade in proliferating cells such as cancer stem cells. 

https://www.ncbi.nlm...les/PMC5378970/

 

 

Note that the curcumin I'm using in the senolytic step also leads to telomere shortening. 

 

 

we found curcumin to be inhibiting telomerase activity and down-regulating hTERT mRNA expression leading to telomere shortening. 

https://onlinelibrar....1002/jcb.24466

 

 

 

While shortening telomeres may seem counter intuitive in view of the desire of so many to lengthen them, the fact is, the telomere clock is only weakly correlated with chronological age while the epigenetic clock tracks it very nicely. I explained the logic of this approach in a previous post, 1197. Longer telomeres result in blocking the natural progression of cellular replacement, which begins with circulating pluripotent cells (VSELs), then to adult stem cells, transit amplifying cells, and finally to somatic cells, which do all the work. Thus shortening telomeres at all points except the first should speed up replacement, resulting in lower epigenetic age. The progression looks like this--

 

VSELs >> SCs >> TACs >> Somatic cells

 

 

Edited by Turnbuckle, 04 June 2020 - 10:07 AM.

[yz69]

consider the standard deviation of trume test, would you say the 0.3 year gain in the latest test could just be noise?

[Turnbuckle]

consider the standard deviation of trume test, would you say the 0.3 year gain in the latest test could just be noise?

 

 

Could be. However I plan to continue tests every 2-4 months, so that possibility should lessen with time. And overall, the two year results are substantial.

[stephen_b]

The slope is fairly consistent over a long baseline. Some tapering off recently. It will be interesting to follow the changes over time.

 

 

Edited by stephen_b, 05 June 2020 - 09:52 PM.

[hsibai]

Thanks Turnbuckle,

 

I presume SA can be substituted by GMS in this protocol. If so, how would that affect dose and timing of the protocol?

 

Thanks!

 

This is an update from post #1149. The only change is with resveratrol. After I added that, I felt there was an acceleration in age reduction, but the drop I saw in the epigenetic test results wasn't significant enough to say it actually improved anything. So best to take it out for now.

 

 

 

 

[Nietzsche]

If you were to follow this protocol using only sulforaphane, what dose would you use?

 

It may be possible to use sulforaphane alone, but I've been content with just cutting it back.

 

 

 

Edited by Nietzsche, 15 June 2020 - 05:36 PM.

[eighthman]

https://www.fightagi...-mice/#comments

 

Someone objects to the mouse model in the comments.  I just ordered the supplements in the protocol.

[ZuoCi]

 

Interesting info here - liver sinusoid endothelial cells (LSEC) gets senescent early(or dont get removed efficiently) and during a study with senolitic treatment they get replaced by fibrotic tissue, this affecting the liver function.

Another problem - with massive senescent cells removal capillary system loses integrity and leads to blood leaking.

 

One could argue that we dont really want too effective senolitic treatment unless this is resolved. Then we probably want something that supresses fibrosis (I believe curcumin shown to decrease liver fibrosis), we need to be sure that we have enough stem cells in a pool and probably need some help with differentiating into correct cell types.

Maybe addition of phospatidylserine would help? I see a small bunch of studies where it helps to differentiate into correct cell types for different environments.

Yes, Tumeric and Taurine are your two top anti-fibrosis supps. 

[platypus]

Excellent, how much younger, if any, would you say you feel after starting these treatments? 

[Turnbuckle]

Excellent, how much younger, if any, would you say you feel after starting these treatments? 

 

The feeling of youth, in my experience, parallels epigenetic age very closely. So does appearance. This was my hypothesis from the beginning, that aging primarily results from the failure of the body to replace aging cells with new cells, and secondarily, the failure of mitochondrial QC to maintain healthy mito populations. Both can be corrected.

[platypus]

The feeling of youth, in my experience, parallels epigenetic age very closely. So does appearance. This was my hypothesis from the beginning, that aging primarily results from the failure of the body to replace aging cells with new cells, and secondarily, the failure of mitochondrial QC to maintain healthy mito populations. Both can be corrected.

Congrats Turnbuckle, you are in the forefront of age-reversal therapies. Your protocols and GDF-11 are the most promising things I've seen in this front since forever. Do you happen to monitor other biomarkers than epigenetic age, like the GDF-11 folks are doing? 

[Turnbuckle]

Congrats Turnbuckle, you are in the forefront of age-reversal therapies. Your protocols and GDF-11 are the most promising things I've seen in this front since forever. Do you happen to monitor other biomarkers than epigenetic age, like the GDF-11 folks are doing? 

 

No. I haven't been following the GDF-11 discussions. Has anyone reported lowered epigenetic age from using it?

[platypus]

No. I haven't been following the GDF-11 discussions. Has anyone reported lowered epigenetic age from using it?

Frankly I don't remember and cannot check right now, but they report lowered reaction-time, which I find astonishing. And then there are reports of gaining more youthful vocal/whistling range, lowered BP etc. 

[eighthman]

https://www.frontier...2020.00515/full

 

Damn !  Confirmation/vindication.......

[ryukenden]

Update on my epigenetic age.
It continues to tick backwards. The last 3 tests from TruMe --

Date Epigenetic age - Chronological age, years
02/2018.... +0.5 (Baseline before treatments)
11/2019.... -13.0
02/2020.... -14.2
04/2020.... -14.5

These treatments presently comprise a stem cell enhancement step once every 10-14 days, and a senolytic + stem cell nutrition step 24 hours later, with another the next day. I've recently added a telomerase shortening agent to the senolytic/nutrition steps, and we'll see how that goes with the next results. This agent is cinnamaldehyde (see the paper below), which is the major component of cinnamon oil. I previously found that telomerase enhancers increased epigenetic age, so its logical to try going the other way.




Note that the curcumin I'm using in the senolytic step also leads to telomere shortening.




While shortening telomeres may seem counter intuitive in view of the desire of so many to lengthen them, the fact is, the telomere clock is only weakly correlated with chronological age while the epigenetic clock tracks it very nicely. I explained the logic of this approach in a previous post, 1197. Longer telomeres result in blocking the natural progression of cellular replacement, which begins with circulating pluripotent cells (VSELs), then to adult stem cells, transit amplifying cells, and finally to somatic cells, which do all the work. Thus shortening telomeres at all points except the first should speed up replacement, resulting in lower epigenetic age. The progression looks like this--

VSELs >> SCs >> TACs >> Somatic cells

Are you thinking of using any other labs apart from Trume to compare? After reading a post at fightaging.org, maybe it is worth tying alternative labs to validate the changes.

[Turnbuckle]

Are you thinking of using any other labs apart from Trume to compare? After reading a post at fightaging.org, maybe it is worth tying alternative labs to validate the changes.

 

Since early 2018 I've used 3 labs. They all appeared to give similar results. I'm presently using Trume, because first they're still in business, and second they're faster and cheaper. Though not as fast and cheap as Osiris Green, so I was sorry to see them go. If you know any others besides those two and myDNAge, let me know.

[yz69]

Here's another company offering DNA methylation clock test for $145

http://epiagingusa.com/product/epiaging-kit/

 

 

[eighthman]

https://www.fightagi...-age-in-humans/

 

Perhaps this is worth a look in the regimen.

[QuestforLife]

https://www.fightagi...-age-in-humans/

Perhaps this is worth a look in the regimen.

I did a 3 1/2 month protocol of AKG and am awaiting a methylation test result. I will post back when I have results.

[Turnbuckle]

This supplement contains a proprietary form of calcium alpha-ketoglutarate, which the FDA considers to be GRAS. The company believes that Rejuvant works by slowing down the rate of age-related DNA methylation...

https://www.fightagi...-age-in-humans/

 

 

This makes sense. Increasing the amount of  methyltransferase or improving its efficiency should lower epigenetic age. Thus if Rejuvant is right about the mechanism, adding it to the SC protocol should multiply the effect.

[Fafner55]

https://www.fightagi...-age-in-humans/

 

Perhaps this is worth a look in the regimen.

 

It looks like a calorie restriction mimic to me.

 

“The metabolite alpha-ketoglutarate extends lifespan by inhibiting ATP synthase and TOR” (2014) https://www.ncbi.nlm...les/PMC4263271/

[yz69]

How does this work?

 

 

Increasing the amount of  methyltransferase or improving its efficiency should lower epigenetic age. 

 

[QuestforLife]

Alpha ketoglutarate is an important intermediate in the Krebs cycle. It is linked to calorie restriction because AMPK increases the enzyme step before AKG (isocitrate dehydrogenase), hence increasing its concentration. AKG has epigenetic effects because it is a cofactor in the TET enzymes, which can remove methylation. This is most likely the mechanism behind its purported effects, but there are many interlinked pathways in energy metabolism and epigenetic control so it's hard to say for sure. It also leads to a reduction in triglycerides and cholesterol because increasing the rate of the Krebs cycle increases the requirement on acetyl coA, which starts drawing on beta oxidation. Therefore there is probably also a link between the effects of AKG and eating stearic acid, which increases beta oxidation. I can dig out a tonme of references if anyone wants them.

Incidentally AKG has been used in salt form (with calcium or magnesium or a mix) for years, I can't see how Rejuvant have a patent. So there is no need to buy their expensive supplement.

[Andey]

I did a 3 1/2 month protocol of AKG and am awaiting a methylation test result. I will post back when I have results.

 

How do you use it? Do you use salt of some kind or plain AKG?

I just started to do the AKG mixed with Guar Gum to supposedly extend digestion period, and take it whatever I remember so few times a day.

[Turnbuckle]

I think the following paper is on point -- 

 

Intracellular α-ketoglutarate maintains the pluripotency of embryonic stem cells

 

The role of cellular metabolism in regulating cell proliferation and differentiation remains poorly understood1. For example, most mammalian cells cannot proliferate without exogenous glutamine supplementation even though glutamine is a non-essential amino acid1,2. Here we show that mouse embryonic stem cells (ESCs) grown under conditions that maintain naïve pluripotency3 are capable of proliferation in the absence of exogenous glutamine. Despite this, ESCs consume high levels of exogenous glutamine when the metabolite is available. In comparison to more differentiated cells, naïve ESCs utilize both glucose and glutamine catabolism to maintain a high level of intracellular α–ketoglutarate (αKG). Consequently, naïve ESCs exhibit an elevated αKG/succinate ratio that promotes histone/DNA demethylation and maintains pluripotency. 

https://www.ncbi.nlm...les/PMC4336218/

 

 

Keeping in mind this study was done with mice, it appears there are two ways to improve pluripotency: with supplemental glutamine, or with α–ketoglutarate. In adults, this should improve the function of VSELs -- the pluripotent backup to adult stem cells and the main target for the protocols of this thread.

 

In vitro, α–ketoglutarate can also promote differentiation of human SCs --

 

α-Ketoglutarate Accelerates the Initial Differentiation of Primed Human Pluripotent Stem Cells

 

Pluripotent stem cells (PSCs) can self-renew or differentiate from naïve or more differentiated, primed, pluripotent states established by specific culture conditions. Increased intracellular α-ketoglutarate (αKG) was shown to favor self-renewal in naïve mouse embryonic stem cells (mESCs). The effect of αKG or αKG/succinate levels on differentiation from primed human PSCs (hPSCs) or mouse epiblast stem cells (EpiSCs) remains unknown. We examined primed hPSCs and EpiSCs and show that increased αKG or αKG/succinate ratios accelerate, and elevated succinate levels delay, primed PSC differentiation.

https://www.ncbi.nlm...les/PMC5023506/

 

 

This result with human cells suggests that α-ketoglutarate should not be taken with the fusion part unless stearic acid is enough to override differentiation. And I expect it will be. I further expect that taking α–ketoglutarate as a mono-therapy would produce lowered epigenetic age in the short term, but may deplete stem cells in the long term.

 

 

Edited by Turnbuckle, 08 July 2020 - 12:16 PM.