2503 replies to this topic

[dlewis1453]

I believe Turnbuckle has used Resveratrol in the fission part of his mitochondria protocol and in the senescent cell clearance portion of his stem cell protocol, but it causes him strong joint pain. 

Edited by dlewis1453, 05 February 2020 - 03:54 PM.

[Turnbuckle]

I believe Turnbuckle has used Resveratrol in the fission part of his mitochondria protocol and in the senescent cell clearance portion of his stem cell protocol, but it causes him strong joint pain. 

 

 

See post #1126 on the previous page.

[Mr Matsubayashi]

Using nicotinamide as a precursor might be better than others because it lowers total blood lipids, including stearic acid. I could only find one study in quick googling that discussed stearic acid and nicotinamide. The effect is seemingly small so maybe not as important. 

https://www.ncbi.nlm.../pubmed/1411265

 

I think stearic acid consumption will promote weight loss. Stearic acid is a signalling molecule to our mitochondria and my guess is that evolution used it to indicate consumption of a fatty meal. When stearic acid is the only lipid consumed the body gears up to process and burn correlating amount of palmitic, oleic, ect. If these are not present it results in weight loss. 

It could also be that stearic acid is poorly absorbed by bile acids if consumed in large amounts and is largely excreted. 

It also seems to increase insulin release. 

https://pdfs.semanti...e4590c4a411.pdf

 

How satiating are your brownies? 

Edited by Mr Matsubayashi, 07 February 2020 - 12:09 AM.

[QuestforLife]

Using nicotinamide as a precursor might be better than others because it lowers total blood lipids, including stearic acid. I could only find one study in quick googling that discussed stearic acid and nicotinamide. The effect is seemingly small so maybe not as important. 

https://www.ncbi.nlm.../pubmed/1411265

 

I think stearic acid consumption will promote weight loss. Stearic acid is a signalling molecule to our mitochondria and my guess is that evolution used it to indicate consumption of a fatty meal. When stearic acid is the only lipid consumed the body gears up to process and burn correlating amount of palmitic, oleic, ect. If these are not present it results in weight loss. 

It could also be that stearic acid is poorly absorbed by bile acids if consumed in large amounts and is largely excreted. 

It also seems to increase insulin release. 

https://pdfs.semanti...e4590c4a411.pdf

 

How satiating are your brownies? 

 

Weight loss on stearic acid has been reported on this thread, by myself and lost69, I believe. The paper 'Dietary stearic acid regulates mitochondria in vivo in humans'  actually states ingesting C18 reduces circulating acyl-carnitines.

[Iporuru]

Using nicotinamide as a precursor might be better than others because it lowers total blood lipids, including stearic acid. I could only find one study in quick googling that discussed stearic acid and nicotinamide. The effect is seemingly small so maybe not as important. 

https://www.ncbi.nlm.../pubmed/1411265

 

You're conflating nicotinamide with nicotinic acid

[Mr Matsubayashi]

You're conflating nicotinamide with nicotinic acid

 

Thanks mate, I was reading too quickly and missed that. 

[Nate-2004]

Turnbuckle, have you seen this?

 

The model systematically underestimates age in tissues from older people. This is seen in all examined tissues but most strongly in the cerebellum and is consistently observed in multiple datasets.

 

https://doi.org/10.1...3059-019-1810-4

[Turnbuckle]

Turnbuckle, have you seen this?

 

 

 

 

https://doi.org/10.1...3059-019-1810-4

 

 

I don't care about systematic effects, as a baseline test two years ago reported my epigenetic age at .5 years above my chronological age. My results since then have drifted consistently lower (except when I experimented with telomerase enhancers), reaching -13 years relative to chronological age late last last year. 

[Turnbuckle]

An experimental protocol for epigenetic age reduction (updated, with threonine restored) —

 

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.

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

  

 

Part 1: Stem cell self-renewal

 

 

Time 0 —

¹Stearic acid — 5-10 g (food grade, in brownie)

 

  

Time 3:00 —

²Sulforaphane — 100 mg

³Liposomal glutathione — 1 g

⁴SAM-e — 500 mg

⁵Resveratrol — 300 mg

 

Time 3:30 —

⁶C60 — 3 mg (in oil)

⁷Amino acids: Threonine — 2-3 g, Lysine — 2 g, Methionine — 1 g, Leucine — 1 g

 

These amino acids should be repeated as necessary every couple of hours, and perhaps days.

 

 

Part 2: Senescent cell replacement (24 hours later)

 

⁸Nicotinamide — 2g, Ribose — 2g

⁹Curcumin (liposomal or phytosomal) — 2g

⁷Lysine — 2 g, Methionine — 1 g, Leucine — 1 g

 

These amino acids should be repeated as necessary every couple of hours, and the entirety of Part 2 may be repeated the next day as well.

 

 

 

Notes:

 

(1) This is for mito fusion, which directs SCs to self-renewal. Food grade stearic acid is a waxy triglyceride with about 50% stearic acid moieties. It has a high melting point and will have very poor availability unless properly prepared. Baking it into brownies is one option: Using a box mix that calls for 1/2 to 2/3 cups of oil, eliminate the oil and add 120 grams of stearic acid flakes or granules, leaving the rest of the recipe unchanged. Mix at room temp using a power mixer, bake according to directions on the box, then divide 3x4 and freeze most of it for later use. Use ½ to 1 brownie for this protocol. With the addition of sulforaphane, I've been using ½. Stearic acid has much longer half life than sulforaphane (11 vs 4 hours), and thus may interfere with subsequent senescent cell replacement, which requires mito fission. It may be possible to use sulforaphane alone, but I've been content with just cutting it back.

 

(2) Sulforaphane penetrates the BBB to produce mito fusion there, while stearic acid is blocked. 50 mg caps can be obtained from Amazon — Thorne Research - Crucera-SGS - Broccoli Seed Extract for Antioxidant Support - Sulforaphane Glucosinolate (SGS)

 

(3) Primary antioxidant.

 

(4) Methyl group donor.

 

(5) Stimulates stem cells and protects from apoptosis

 

(6) UCP2 blocker. A teaspoon of commercially available C60 oil.

 

(7) These can be used in caps or tablets, though I use a commercial blend of essential amino acids here, adding extra lysine, methionine and leucine. Pluripotent cells have a special need for these three. I would spread out the doses as indicated, to avoid nausea. Note that I deleted threonine in the last update, but I got negative results by leaving it off. This became increasingly apparent after a few weeks, thus I restored it.

 

(8) Used together, nicotinamide and ribose rapidly increase NAD+, which drives mitochondrial fission, required for senescent cell apoptosis and replacement.

 

(9) Senolytic

 

 

 

[dlewis1453]

 

Note that I deleted threonine in the last update, but I got negative results by leaving it off. This became increasingly apparent after a few weeks, thus I restored it.

 

 

 

 

Thanks for the updated protocol Turnbuckle. Can you expand a bit on the effects you noticed when you ceased and then resumed your threonine supplementation? 

[Turnbuckle]

Thanks for the updated protocol Turnbuckle. Can you expand a bit on the effects you noticed when you ceased and then resumed your threonine supplementation? 

 

 

I often feel flu like symptoms during the senolytic phase of a treatment, but this was worse. It included joint pain and seemed to build over subsequent days, even without senolytics. Several grams of threonine put an end to it very quickly. 

[QuestforLife]

I often feel flu like symptoms during the senolytic phase of a treatment, but this was worse. It included joint pain and seemed to build over subsequent days, even without senolytics. Several grams of threonine put an end to it very quickly. 

 

Are you sure it's not just a requirement for glycine? Threonine a precursor to glycine, and we know human ESCs don't have the requirements for threonine murine ESCs do (I posted references to this previously). Might be worth a future experiment (glycine is also much cheaper than threonine).

 

One other thing, how did you arrive at curcumin as a senolytic?

[Turnbuckle]

Are you sure it's not just a requirement for glycine? Threonine a precursor to glycine, and we know human ESCs don't have the requirements for threonine murine ESCs do (I posted references to this previously). Might be worth a future experiment (glycine is also much cheaper than threonine).

 

Unlike threonine, glycine is nonessential, so developing a glycine deficiency seems unlikely. As for the cost, threonine powder is around 2 cents a gram based on Amazon's price for 1 kg, while glycine is slightly more. Both are dirt cheap.

 

It's worth doing a test, however, as you say.

 

One other thing, how did you arrive at curcumin as a senolytic?

 

 

Curcumin is known to upregulate p53, and p53 is induces apoptosis. Combine that with NAD+ induced mito fission (necessary for apoptosis) along with the availability of active stem cells, and expect enhanced turnover of senescent cells.

Edited by Turnbuckle, 19 February 2020 - 10:44 AM.

[QuestforLife]

Unlike threonine, glycine is nonessential, so developing a deficiency seems unlikely. As for the cost, threonine powder is around 2 cents a gram based on Amazon's price for 1 kg, while glycine is slightly more. Both are dirt cheap.
 
It's worth doing a test, however, as you say.

 

Glycine is what is known as conditionally essential. We can make it in vivo, but not enough, and increasingly with age have a deficit. (Threonine is expensive in the UK whereas Glycine is cheap, for some reason, so for me there is added reason to do the experiment).

 

Curcumin is known to upregulate p53, and p53 is induces apoptosis. Combine that with NAD+ induced mito fission (necessary for apoptosis) along with the availability of active stem cells, and expect enhanced turnover of senescent cells.

 
I always thought curcumin was lacklustre as a senolytic, but that was no doubt testing it alone; combining it with NAD+ makes sense. I assume you have found it is effective by trial and error over other compounds? I seem to remember you used butyrate before.

[granmasutensil]

 

Unlike threonine, glycine is nonessential, so developing a glycine deficiency seems unlikely. As for the cost, threonine powder is around 2 cents a gram based on Amazon's price for 1 kg, while glycine is slightly more. Both are dirt cheap.

 

It's worth doing a test, however, as you say.

 

 

Curcumin is known to upregulate p53, and p53 is induces apoptosis. Combine that with NAD+ induced mito fission (necessary for apoptosis) along with the availability of active stem cells, and expect enhanced turnover of senescent cells.

 

 

A suggestion. What about killing two birds with one stone and replacing SAMe for TMG as a methyl donor, TMG also supplies glycine? Perhaps it's cellular hydration properties would also help.

[Turnbuckle]

Glycine is what is known as conditionally essential. We can make it in vivo, but not enough, and increasingly with age have a deficit. (Threonine is expensive in the UK whereas Glycine is cheap, for some reason, so for me there is added reason to do the experiment).

 

 
I always thought curcumin was lacklustre as a senolytic, but that was no doubt testing it alone; combining it with NAD+ makes sense. I assume you have found it is effective by trial and error over other compounds? I seem to remember you used butyrate before.

 

 

Living in the US has a definite advantage when it comes to availability of supplements. I always try to find the least expensive approach, but being US centric, it may not be the least expensive for everyone. As for cucurmin, it is not a strong senolytic. I originally used other things to get a decent effect, but eventually I discovered I didn't need them, and sometimes--after an SC treatment--I didn't need anything but NAD+ and SC nutrition. This is based on subjective effects as I haven't broken out that part of it for epigenetic tests. As you probably know, epigenetic changes are slow and so are the tests, taking 5 weeks or more to get them back.

[ambivalent]

You had eliminated lack of threonine as the cause of the joint pain, not inclusion of resveratrol*?

 

Sounds as though this might be a build of uric acid which the glycine via threonine off-set. You may not recall the problems I had a few years ago through high dosing NR which led to big spikes in uric acid and kidney problems.

 

It should be in the thread somewhere but it may well have been your suggestion that it was excess d-ribose causing the uric acid spike. Just a quick search:

 

Fenstad et al. ¹⁵⁾found significantly higher levels of uric acid (which were, however, still in the normal range) following oral administration of 10 g D-Ribose when compared to the dose of 2 g at 30 and 60 min, no changes in serum uric acid concentrations were reported by Thompson et al. ¹⁶⁾ following ingestion of 10 g d‐ribose and in the study of Gross et al. ¹⁷⁾, during or after treatment with 27–89 g d‐ribose. Gastrointestinal symptoms were noted at single intakes of around 70 g ¹⁸⁾.

 

Needless to say I wasn't anywhere near these levels but I was pretty sick on taking up to 7gms of NR, and some N+R. When tested several months later after improving (lots of cherry juice) my uric acid levels were just in normal range.

 

Anyhow, glycine seems to lower uric acid:

 

https://www.ncbi.nlm...les/PMC6471320/

 

It appears to have been an old treatment for the condition:

 

https://ard.bmj.com/...9/1/38.full.pdf

 

 

 

 

*Incidentally, I suffered badly from joint pain when taking trans-ptero 

 

 

Edited by ambivalent, 19 February 2020 - 12:18 PM.

[Turnbuckle]

You had eliminated lack of threonine as the cause of the joint pain, not inclusion of resveratrol*?

 

Sounds as though this might be a build of uric acid which the glycine via threonine off-set. You may not recall the problems I had a few years ago through high dosing NR which led to big spikes in uric acid and kidney problems.

 

It should be in the thread somewhere but it may well have been your suggestion that it was excess d-ribose causing the uric acid spike. Just a quick search:

 

Fenstad et al. ¹⁵⁾found significantly higher levels of uric acid (which were, however, still in the normal range) following oral administration of 10 g D-Ribose when compared to the dose of 2 g at 30 and 60 min, no changes in serum uric acid concentrations were reported by Thompson et al. ¹⁶⁾ following ingestion of 10 g d‐ribose and in the study of Gross et al. ¹⁷⁾, during or after treatment with 27–89 g d‐ribose. Gastrointestinal symptoms were noted at single intakes of around 70 g ¹⁸⁾.

 

Needless to say I wasn't anywhere near these levels but I was pretty sick on taking up to 7gms of NR, and some N+R. When tested several months later after improving (lots of cherry juice) my uric acid levels were just in normal range.

 

 

 

2 grams of ribose isn't the cause, and it happened with and without resveratrol. 

[ambivalent]

Well, be that as it may: other symtoms were tingling, neruopathy and foot pain (which lasted months after cessation, likely nerve damage). I wasn't alone in reporting those symptoms from NR (others at lower doses). I became extremely sensitive to any dose. 

Edited by ambivalent, 19 February 2020 - 12:46 PM.

[Turnbuckle]

Well, be that as it may: other symtoms were tingling, neruopathy and foot pain (which lasted months after cessation, likely nerve damage). I wasn't alone in reporting those symptoms from NR (others at lower doses). I became extremely sensitive to any dose. 

 

 

This was not a daily dose, but maybe once per SC treatment--once a week or so of 1-2 g each of N+R. I never experienced the symptoms I noted above from that before. If you experienced neuropathy from NR or other NAD+ raisers, it might be from large amounts of ribose, or it might derive from excessive mitophagy and lower mitochondrial mass, and subsequent apoptosis. Especially if you were to take it every day. For instance--

 

enhanced levels of mitophagy can lead to excessive removal of mitochondria, loss of cardiac myocytes, and development of heart failure

https://www.ncbi.nlm...les/PMC3538875/

 

 

Extended periods of all fission is not good, just as extended periods of all fusion is not good.

[ambivalent]

Well certainly I experienced excessive fatigue and endurance, heightened alertness and extreme brain fog, as well what appeared to be visually rejuvenating effects til I crashed - which appears consistent with that finding, except the latter which I put down to raised NAD+. As you infer its the d-ribose which is known to raise glycated haemoglobin and caused the nerve damage. Still it seemed a lot of damage for not really that much d-ribose. All NAD precursors I now take sublingually. 

 

 

[ambivalent]

 I mentioned before somewhere it felt as though the symmetric stem cell differentiation was a response to a fasted state as Longo demonstrated there was stem cell activation during the feeding phase to replace the fasting induced metabolised organelles. So it would seem that the increase in stem pool size during fusion is in preparation for the post-fasting state. On fasting and mitochondrial fusion:

 

https://www.ncbi.nlm...es/PMC4231308/ 

 

These findings show that adaptive mitochondrial fusion protects metabolically challenged mitochondria.

 

This would seem at points a danger with the ketogenic diet and explains why so many feel highly energised on it. It would also be presumably wise for regular fasters too, to induce states of fission between fasts.

 

As for Turnbuckle's protocols is fasting practical or as effective ?  Fusion might take too long to induce after fission via fasting than stearic acid. Could be worth a go for those worried about palmitric acid in the commercial food grade stearic acid.

Edited by ambivalent, 20 February 2020 - 11:38 PM.

[Empiricus]

Well, be that as it may: other symtoms were tingling, neruopathy and foot pain (which lasted months after cessation, likely nerve damage). I wasn't alone in reporting those symptoms from NR (others at lower doses). I became extremely sensitive to any dose. 

 

You had foot pain as well? In addition to foot pain (arches of feet), I had also experienced kidney pain while I was taking NR. 

 

However, kidney pain I attributed entirely to olive leaf and hydroxytyrosol (an olive polyphenol).  It could well be the NR was a factor in my kidney pain.  

[ambivalent]

Yes, I did, there were other cases reporting it too in the NR personal experiences thread. Its worth checking H1abc (ribose) and uric acid (nicotinamide) iirc. There are links in a few other threads. These were early warning signs which I ignored (about two days before suffering acute kidney pain I felt amazing). I switched to sublingual NMN and recovered well (vision improved too after becoming poor too), I will try again high doses of NR but likewise under the tongue. Still a little cautious of N+R in TB's protocols but of course its only in blasts.

 

One additional point, while fasting induced mito-fusion may well be different from the stearic-acid induced state it would seem as though sustained mito-fusion is not an artificial and damaging cellular condition (or at least recoverable) - perhaps, though, there are fission breaks during a sustained fast as energy levels can rise and fall. 

 

 

[Turnbuckle]

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

 

To recap. 

 

Test date...Epigenetic age - chronological age

2/2018 ... +.5 years (This was baseline, before treatments)

6/2018 ... -11

 

I began to use telomerase stimulants at this point, and my epigenetic age began to increase. When I stopped telomerase stimulants, epigenetic age stabilized and slowly began to fall with continued SC treatments. Cycloastragenol and astragalus extract screwed up my age regression for a year.

 

12/2019 ... -13

2/2020 ... -14.2

 

Edited by Turnbuckle, 02 March 2020 - 06:00 PM.

[dlewis1453]

Great result! Thanks for sharing as always. 

[ryukenden]

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

 

To recap. 

 

Test date...Epigenetic age - chronological age

2/2018 ... +.5 years (This was baseline, before treatments)

6/2018 ... -11

 

I began to use telomerase stimulants at this point, and my epigenetic age began to increase. When I stopped telomerase stimulants, epigenetic age stabilized and slowly began to fall with continued SC treatments. Cycloastragenol and astragalus extract screwed up my age regression for a year.

 

12/2019 ... -13

2/2020 ... -14.2

Great results. 

[Nate-2004]

All good things to know!! So how many SC cycles/treatments have you gone through and how many were just like the above protocol with the senescent cell treatment a day after?

 

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

 

To recap. 

 

Test date...Epigenetic age - chronological age

2/2018 ... +.5 years (This was baseline, before treatments)

6/2018 ... -11

 

I began to use telomerase stimulants at this point, and my epigenetic age began to increase. When I stopped telomerase stimulants, epigenetic age stabilized and slowly began to fall with continued SC treatments. Cycloastragenol and astragalus extract screwed up my age regression for a year.

 

12/2019 ... -13

2/2020 ... -14.2

 

All good things to know!! So how many SC cycles/treatments have you gone through and how often and how many were just like the above protocol with the senescent cell treatment a day after? 

Edited by Nate-2004, 02 March 2020 - 11:40 PM.

[Andey]

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

 

  How long does it takes Trumelabs to run a sample? I recently got it delivered it to them and dying of curiosity)

 

BTW I recall reading that their between samples variance(same person, several tests during one day) is 3 month max so your 1.2 years is definitely above it. 

[Turnbuckle]

  How long does it takes Trumelabs to run a sample? I recently got it delivered it to them and dying of curiosity)

 

 

 

This last TruMe test took 17 days after receipt -- about 3 times faster than mydnage.