2503 replies to this topic

[Turnbuckle]

Update on my epigenetic age.

 

This is an update on my personal experiment with age reversal. The previous update was in post 1441. This chart includes a test by a new lab — EpiAging. These labs use different markers and methodology, so they can't be expected to be the same even though both use saliva. The EpiAging sample was taken within ten minutes of the TruMe sample on 10/2020. The average of the two tests shows an age reversal of -20.4 years.

 

Date              Test   Epigenetic age - Chronological age, years

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

11/2019*.... -13.0

02/2020*.... -14.2

04/2020*.... -14.5

06/2020*.... -13.0 (+ cinnamon powder)

08/2020*.... -11.3 (+ cinnamaldehyde & eugenol)

 

10/2020*.... -22.6 (+ Arginine AKG, 2-3 grams)

10/2020**... -18.1 (+ Arginine AKG, 2-3 grams)

 

 

* TruMe test

** EpiAging test

 

 

Edited by Turnbuckle, 15 December 2020 - 08:46 PM.

[kurt9]

This is clearly an effective age reversal technique. I guess the next step beyond it would be invivo partial cellular reprogramming. Maybe we can have that in the next 5 years or so.

[kurt9]

I've a question for you guys. Can this stem cell renewal protocol be used to overcome auto-immune conditions, like those associated with vaccines?

[Fafner55]

Turnbuckle,

Do you suggest taking your protocol's supplements in the morning or evening? And related, should low dose (3 mg) melatonin be taken before bedtime as part of the protocol to better regulate the circadian rhythm and stem cell state, particularly in older people.

 

The circadian rhythm plays a role in stem cell activation, maintenance and differentiation.

1. "Adult stem cell maintenance and tissue regeneration around the clock: do impaired stem cell clocks drive age-associated tissue degeneration?" (2018) https://link.springe...0522-018-9772-6
2. "Circadian clock-mediated control of stem cell division and differentiation: beyond night and day" (2014) https://dev.biologis...ent/141/16/3105

 

During times corresponding to early morning, differentiation pathways (of in vitro epidermal keratinocyte stem cells) were high, whereas in the evening pathways corresponding to DNA replication and cell division predominated.

"Human epidermal stem cell function is regulated by circadian oscillations" (2013) https://www.cell.com...49?showall=true

 

[Turnbuckle]

Turnbuckle,

Do you suggest taking your protocol's supplements in the morning or evening? 

 

See post 1465 on the previous page.

[userCK]

@Turnbuckle - Can ISRIB be incorporated into this or your Alzheimer's protocol?

Anyone knows where to get legit ISRIB?

 

[aribadabar]

@Turnbuckle - Can ISRIB be incorporated into this or your Alzheimer's protocol?

Anyone knows where to get legit ISRIB?

These guys have been around for a while supplying many other exotic compounds. Sigma's pricing is beyond reach for mere mortals.

[ambivalent]

On hair:

 

Before taking c60 in the summer of 2013 I was frequently plucking out grey eyebrow hairs, there was quite a bit of friendly fire  (notieceable deforrestation). Post c60 though and to this day, it is a rare occurrence and the foliage is pretty dense. In addition when taking c60 (I do so infrequently at quite large doses) there is a real spurt, with the growth bordering on running out of contol (it gets trimmed). I recall one poster possibly Adamzski proudly showing off a 2 inch c60 induced eyebrow hair.

 

Pre c60, I had bald shins and at the top of my legs, which I put down to zinc deficiency. The hair returned post c60 it went unnoticed until taking me quite by surprise one day. This  resulted from c60, I'm sure, but also may have occurred later due to NAD increases too. It is a problem which reccurs, but it was a very impressive effect and has been correlated with large doses of c60 and quite quick to take effect.

 

'Other hair' - seemingly some de-greying from c60 compared to prec-60.

 

Head hair - my suspicion is that c60 may have held off greying, I've been lucky in that regard but not lucky in loss of this hair - I have a very receded hairline.

 

Facial hair - there was a time when I paid close attention, it seemed as though I was winning the war a small amount of grey in 2013 which i seem to hold off and sometimes it 'I thought' reduced. Anyhow 2017 was a year of bad experimentation and I became unwell with kidney problems which seemed to accelerate aging during that period. And the grey increased. For now I have aceepted defeat.

 

Now curiously, when i started taking senolytics most notably large doses of fisetin - I observed regrowth of hair around the temple. It was also PMed that a guy taking senolytics and another compond had some temple hair regrowth although not indicated whether he had receding hair elsewhere. Temple hair is of course slow growing and I assume limited. 

 

So it terms of regrowth and de-greying there does appear traction with c60 and other supplements/protocols with hair of limiting length. So what differentiates those hair cells from those of non-limiting length? 

 

 

[ambivalent]

Also I took two protocols of senolytics predominantly fisetin (5g+). It took a couple of days but i noticed a weakening in the knees but mild compared to the first dose 2 years prior, possibly an indication of the repletion rate of senescent cells. One arthritic knee was especially bad after the first time but was quickly stronger after recovery. There were the ususal subjective improvements in skin but nothing I could prove. I later followed up with a stearic acid/ c60 protocol. 

 

One unmistakeable effect of the large dose of fisetin, which I had noticed before but did't report before out of sensitivity but do so now mostly for entertainement purposes - that which is usually white, was very yellow (for about 24 hours at a guess)

[ambivalent]

Only today have I seen the unpromising recent c60 study on mice - this study came back to me: 

 

https://pubmed.ncbi....h.gov/21410319/

 

it does seem reasonable to hypothesize this is related to with production of stem cells. And we know that fasting induces mitochondrial fusion amd also know that fasting increases stem cell proliferation.

 

But what about the mice, is there something to take from this?

 

 

Mice appear to have some latent ability ro maintain mitochondrial performance under starvation without fusion (prevented through KO HDAC6) - the cost, increased ROS production

Edited by ambivalent, 27 December 2020 - 11:55 PM.

[Rocket]

I wonder when the people restoring their pools ofmstem cells who are over 40 last had their prostates looked at in a psa test. As some people here have reported feet growth, the same effects will be seen in the prostate as elevated psa values and the development of urinating issues that comes with bph. As is known from scientific literature people with acromegaly have far more cases of bph than normal people because of higher hgh that stimulates stem cells in their prostates.

If there are no rising levels of psa in men over 40 on this treatment then it likely doesn't work and isn't stimulating any stem cell renewal.

[bladedmind]

Grateful for your work.  Application of earlier protocols rescued me from eight years of post-exertional malaise (not chronic fatigue, but 24+ hours of exhaustion after strenuous exercise longer than 15 minutes). 

 

Questions about the latest update.          

 

1.  “Lysine and methionine should be repeated as necessary every few hours.”  Just do it every few hours, or take it when you feel down, or something else? 

 

2.  Azithromycin, quercetin add-ons.  Do this early in a single treatment, or early in a string of treatments, or later, or just experiment observing subjective reaction?

 

3.  If I remember correctly, an earlier protocol was not supposed to exceed ten or so repetitions.  But the current protocol suggests 2 or 3 times a month.  Indefinitely?   I’m vague about C60 and the boom and bust effects it had – does this protocol overcome that problem?    

 

Thank you.

[timedilation]

I just finished reading through this thread.  This is all very interesting stuff -- I am pretty surprised there hasn't been more animal research trying ideas like this one to increase stem cell reserves.

I wonder how this protocol might tie into the growing usage of healing peptides like BPC-157, TB-500, GHK-Cu and the like.  Could they be stimulating mass asymmetric stem cell division to promote fibroblast production and angiogenesis?  Could frequent users (even young ones) be depleting their stem cell pools over time?  It would explain why we haven't evolved large endogenous sources of peptides to heal all our wounds.  If so, could the use of this protocol between peptide cycles increase their effectiveness?  I am eager to hear if anyone has relevant studies or thoughts on the matter.

[Rocket]

I just finished reading through this thread. This is all very interesting stuff -- I am pretty surprised there hasn't been more animal research trying ideas like this one to increase stem cell reserves.

I wonder how this protocol might tie into the growing usage of healing peptides like BPC-157, TB-500, GHK-Cu and the like. Could they be stimulating mass asymmetric stem cell division to promote fibroblast production and angiogenesis? Could frequent users (even young ones) be depleting their stem cell pools over time? It would explain why we haven't evolved large endogenous sources of peptides to heal all our wounds. If so, could the use of this protocol between peptide cycles increase their effectiveness? I am eager to hear if anyone has relevant studies or thoughts on the matter.

In short, no.
People with acromegaly don't run out of stem cells at 33 after all..... I in my 40s didn't run out of stem cells gaining 60 pounds of muscle. I've been running BPC-157for a long time now and I heal just fine.

[timedilation]

In short, no.
People with acromegaly don't run out of stem cells at 33 after all..... I in my 40s didn't run out of stem cells gaining 60 pounds of muscle. I've been running BPC-157for a long time now and I heal just fine.

 

Well, of course it wouldn't happen over night -- asymmetric stem cell division only occasionally results in a net loss.  Far more likely is that there would be a gradual but accelerated decline in wound healing over a (probably long?) period of time.  It is interesting that you bring up acromegaly though, as it shortens the lifespan by about 10 years if left untreated.  That actually lines up with this idea pretty well, though obviously GH is much different from BPC. 

 

There is a whole lot we don't know about the interaction of VEGF/FGF/HGH/other growth factors with our stem cells and how fibroblasts fit into to all of that.  It is certainly possible that the effects are minimal, but it would be nice to have more info.  If symmetric stem cell division could increase the potency of peptides, this protocol could become even more important.

[Rocket]

A gradual accelerated decline in wound healing occurs naturally in humans with ageing for various reasons.... The ecm, hgh, signaling... acromegally patients live a normal span when they get treated and their early deaths when not treated aren't the result of stem cell loss. They are prone to bph and cancers as an example.

Until you can quantify stem cells numerically all this is pie in the sky conjecture.

I've gained 60 pounds of muscle bone and other tissues in my early 40s. Assuming you have a fixed N of stem cells in your life, I must be running on empty with all that growth in midlife and I have no issues healing. I healed a completely broken foot well enough in 5 weeks to resume squats and a sprained shoulder in about 3 weeks to resume heavy lifting. As long as cells can differentiate how can you run out? Its a hypothesis and to assume its true is unwise that c60 will deplete your stem cells. As feynman said "shut up and calculate." show me the data.

[Turnbuckle]

 Assuming you have a fixed N of stem cells in your life,

 

That's a false assumption. The numbers of viable stem cells typically decline with age. 

There is increasing evidence that the aging process can have adverse effects on stem cells. As stem cells age, their renewal ability deteriorates and their ability to differentiate into the various cell types is altered. Accordingly, it is suggested aging-induced deterioration of stem cell functions may play a key role in the pathophysiology of the various aging-associated disorders.

 

https://www.ncbi.nlm...les/PMC5316899/

 

 

And from the same paper --

 

Adult stem cells, also known as somatic stem cells, are found throughout the body in every tissues and organs after development and function as self-renewing cell pools to replenish dying cells and regenerate damaged tissues throughout life[16]. However, adult stem cells appear to age with the person. As stem cells age, their functional ability also deteriorates[12,17]. Specifically, this regenerative power appears to decline with age, as injuries in older individuals heal more slowly than in childhood. 

 

 

Adult stem cells thus appear to have a finite life, but by pushing viable stem cells to proliferate (self-renew) rather than differentiate, the numbers of viable stem cells can be increased (though this is limited by the body's homeostatic mechanisms). This bias to proliferation is achieved by manipulating mitochondrial morphology, as discussed in the OP. C60 is known to stimulate stem cells, but depending on mito morphology (fission vs fusion) you will get differentiation or proliferation, with the normal bias set to 80% differentiation. And differentiation is how stem cell pools get depleted to begin with.

 

A chart illustrating the age-associated decline of one type of stem cell is from this page.

Attached Files

•  Decline of SCs with age.PNG   307.23KB   0 downloads

[timedilation]

It appears mTOR manipulation may have a place in this protocol.  I came across a study "mTORC1 Activation during Repeated Regeneration Impairs Somatic Stem Cell Maintenance," which indicates that mTORC1 signaling is responsible for stem cell differentiation and depletion over time.  

 

https://pubmed.ncbi....h.gov/29220665/

 

A few quotes:

 

 

 In human embryonic stem cells, activation of S6K by mTORC1 has been reported to induce differentiation (Easley et al., 2010), and reduction of mTORC1 activity in Paneth cells (ISC support cells) under dietary restriction promotes ISC maintenance and proliferation through a non-autonomous mechanism (Yilmaz et al., 2012).

 

However,

 

 

mTORC1 also appears to play a critical role in the activation of SC proliferation during regenerative episodes in various systems: short term induction of AKT promotes HSC pool expansion (Kharas et al., 2010), while in a mouse model of muscle injury, mTORC1 activation is necessary to allow rapid proliferation of muscle SCs (MuSCs) by promoting a switch from the G0 to a mTORC1-dependent ‘Galert’ state (Rodgers et al., 2014). During hair regeneration, mTORC1 signaling promotes hair follicle SC activation by repressing BMP signaling.

 

The key part:

 

 

Critically, mTORC1 activation is a likely cause of BC loss in aging mice, as long-term treatment with the mTORC1 inhibitor Rapamycin is sufficient to prevent BC loss and restore BC numbers in aging mice. Our data indicate that this loss may be caused by mTORC1-dependent differentiation of BCs. These effects are conserved in the muscle, as activation of mTORC1 promotes reprogramming of the MuSC transcriptome towards a differentiation signature, and the age-dependent loss of MuSCs can be prevented by limiting mTORC1 activity genetically in these cells.

 

Edited by timedilation, 06 February 2021 - 12:08 AM.

[userCK]

1. Azithromycin and Quercetin are in part 2 (meaning, to be taken 24 hours or later)?

2. Other than Azithromycin, can I take double dose of everything, for example, 4 grams of Niacinmide instead of 2g; and other ingredients? I ask because many studies have shown that health supplements are often under-dosed so to be sure I plan to take all ingredients from two different brands.

 

 

[userCK]

Folks, I'm in Poland now and I have not been able to find Threonine as a standalone product. I found some Amino Acid complex that have several amino acids (on iHerb.EU) - is that acceptable in this protocol? One cap has only 112mg of Threonine so I'd need to take like 20caps for 2+grams.

 

[JamesPaul]

Can you order via mail?  I live in the U.S. but have purchased food supplements from the U.K.

 

http://vitamins-supp...tnSearch=Search

[userCK]

Can you order via mail?  I live in the U.S. but have purchased food supplements from the U.K.

 

http://vitamins-supp...tnSearch=Search

With Brexit, I think it is difficult to get stuff from the UK. Can't I just eat a bunch of eggs? They have all the essential amino acids necessary and more.

[Turnbuckle]

I designed this protocol for myself and other Americans who can easily get the ingredients. If you in the UK or elsewhere and can't get something, leave it off. The critical stuff for proliferation is C60, a fusion agent, methionine and lysine, and AAKG (or AKG).

[pamojja]

Folks, I'm in Poland now and I have not been able to find Threonine as a standalone product.

 

Here you go: https://www.sunday.de/threonin/ use deeple.com for excellent translation from German.
 

[userCK]

I designed this protocol for myself and other Americans who can easily get the ingredients. If you in the UK or elsewhere and can't get something, leave it off. The critical stuff for proliferation is C60, a fusion agent, methionine and lysine, and AAKG (or AKG).

Thanks!

And higher than the dosages you suggested is OK? For example, in Mito protocol, I doubled my doses by using different brands of same ingredient (incase a brand was dud or underdosed).

 

Here you go: https://www.sunday.de/threonin/ use deeple.com for excellent translation from German.
 

Thanks!

[nadaepeu]

With Brexit, I think it is difficult to get stuff from the UK. Can't I just eat a bunch of eggs? They have all the essential amino acids necessary and more.

I guess you can try this https://www.ebay.co....872.m2749.l2649

[userCK]

I guess you can try this https://www.ebay.co....872.m2749.l2649

Thanks. ^ doesn't ship to Poland where I'm because of Brexit, I suppose. I ordered from Sunday.de that was posted - they ship to Poland - all amino acids were available there and they're all Vegan source. Protein is protein, so I suppose it's fine.

[simon007]

Hi Turnbuckle,

 

Thanks for sharing this amazing research!

Maybe you should consider writing a case study paper about yourself.

 

What do you notice from having 20 years taken off from your epigenetic age?

Stuff like vision, VO-max, exercise, gray hair etc..

 

What do you think caused the 10 year jump?

What is in your opinion the best fusion agent?

 

Thanks and regards,

 

Simon

Edited by simon007, 17 February 2021 - 08:25 AM.

[yz69]

Hi Turnbuckle,

 

Do you think AKG is good in the senolytic part of SC protocol?

 

Thanks!

 

 

[Turnbuckle]

Hi Turnbuckle,

 

Do you think AKG is good in the senolytic part of SC protocol?

 

Thanks!

 

 

It won't hurt, but it likely won't help much either. Demethylase will do its thing during division, and SC division is forced by C60 during proliferation, so the time frame is predictable, but in differentiation for senescent cell replacement, the timing is dependent on senescent cell apoptosis and paracrine signaling. The process could be lengthy, perhaps taking days. So AKG or AAKG would be too fast. And it would be a duplicated effort, I expect, as once you’ve expanded your SC pools and cleaned them up of aberrant marks, it’s unlikely that doing it again for differentiation would help that much.