2286 replies to this topic

[Empiricus]

This post of potential interest to those having negative reactions to nicotinamide/ nicotinamide plus ribose.  Doing the mito profile using N+R has caused nighttime wakefulness for me:  difficulty falling asleep and staying asleep.  Despite some sleep deprivation I've still found the protocol beneficial and have done it periodically over the last several years.  Substituting apigenin for N+R has fixed the sleep problem, and in fact this modified protocol seems to be something that might improve my sleeping generally.  I've been using 500 mg of a liposomal formulation from which I get a fission 'feel' more intense than from N+R.  I've experimented with this apigenin on its own, and I feel the effects of it for about 48 hours.  I realize that studies have shown conflicting results re the half-life of apigenin, ranging from 12 hours to 90+ hours.  I expect it varies among individuals.  Based on this experience, I've modified the protocol to a three day cycle.  First day Apigenin +AKG + PQQ.  Second, AGK plus PQQ.  Third day AKG, PQQ, DHM.  I've completed three passes.   First pass, day 1 and day 2 felt like the kind of fission days I'm used to after an absence from the protocol, sort of confirming my impression that apigenin still circulating to effect.  Fusion day felt like a fusion day, IE, pleasant release from the fission feeling.  Second and third pass, fission days much better (less relative lift during fusion day) perhaps mitigated by better quality sleep.

 

I've just embarked on the protocol having modified rarefried's version as follows:  First day AKG + PQQ + 150 mg (non-liposomal) apigenin + 350 mg of niacinamide for fission; second day AKG + PQQ + 350 mg niacinamide; and on the third AKG + PQQ + 3 g GMS + 800 mg DHM. 

 

I'm curious whether "less is more" holds true for niacinamide, and I don't think it will hurt to toss the riboside.   

Edited by Empiricus, 29 September 2022 - 12:20 PM.

[PAMPAGUY]

The first time I took GMS, it was about 8 grams, and it was frightening the way my BP shot up. If I was ever to kill myself with my experiments, that would have done it. Stearic acid triglycerides will raise my BP as well, but that is so slow the body can adjust to it. So use no more than one gram of GMS, at least initially. And if that is still a problem, switch entirely to DHM. I've taken large amounts of DHM and haven't seen a BP problem. Another reason I use such small amounts is to prevent residual fusion agents from interfering with the fission treatment the next day.

 

Turnbuckel, thanks for feedback.  So the culprit is GMS, will stick to 1 gram, and monitor B/P.  Also, have been using AAKG instead of AKG.  Had a hard time finding AKG in Europe.  I assume 4 gms of AAKG will do the trick of acting quickly.

[Learner056]

Not so fast, I would not be so sure to attribute high blood pressure on GMS/Stearic alone:  It would be important to know the possible underpinnings.  Like we used to say, if you cannot explain it then you are just winging it. 

 

a) I am taking DHM, I am not taking Stearic (though for transparency, I do take cocoa butter which has natural stearic in it, but that should not matter in my view).  And my BP has never been high until now.  Obviously, something is going on that previously was not.  And if you cannot explain that, then .... 

 

b) While I do read good information on DHM.  But I am concerned as my muscles have been twitching (like something happening to them).  Ya Ya I know I can take Mg, but something is going on, I know my body deeply.  I looked up, research papers say: DHM increases 'slow twitch muscle fibers' while decreases 'fast twitch muscle fibers'.  Apparently to author that is good, but why would "fast be bad" while "slow be good"?

 

Ref: DHM Increases MyHC1 (slow-twitch), while decreasing MyHC2 (fast-twitch), https://pubs.rsc.org...2/FO/D2FO02173K

 

So the culprit is GMS

 

Edited by Learner056, 02 October 2022 - 10:07 PM.

[Turnbuckle]

It is quite possible that stearic acid is not the culprit for hypertension, but palmitic acid. The fatty acids of food grade stearic acid triglycerides average around 50% stearic acid and 50% palmitic acid. The same is true for the monoglyceride. Half of what is called "stearic acid" is palmitic acid in commercially available products, including GMS. There are three types of GMS according to the USP, where Type I has 40-60% stearic acid, and Type III has 80-99% stearic acid.

 

If the type is not specified, it is likely Type I, as it is the most inexpensive. Cholesterol esters of stearic acid lower BP, while cholesterol esters of palmitic acid raise BP --

 

 

Using stepwise linear regression, we determined that each SD increase (1.9%) in the serum level of cholesterol ester palmitoleic acid (16:1) was associated with a systolic pressure increase of 3.3 mm Hg (95% confidence interval, 0.9 to 5.6 mm Hg) ... Serum level of cholesterol ester stearic acid (18:0) was inversely associated with diastolic pressure: each SD increase (0.2%) was associated with a decrease of 1.4 mm Hg (95% confidence interval, −2.5 to –0.2 mm Hg). https://www.ahajourn...01.HYP.27.2.303

 

 

Thus Type III should produce a lower BP, while Type I appears to raise it. 

DHM has been found to lower hypertension in rats, eg: this paper.

 

Edited by Turnbuckle, 03 October 2022 - 03:33 AM.

[johnhemming]

An interesting point about Palmitic and Stearic acid. Which encouraged me to find this paper:

 

https://pubmed.ncbi....ih.gov/8596480/

 

"Different effects of palmitic and stearic acid-enriched diets on serum lipids and lipoproteins and plasma cholesteryl ester transfer protein activity in healthy young women

 

Abstract

The effects of palmitic and stearic acid-enriched diets on serum lipids, lipoproteins, apolipoproteins (apo) A-I and B, and plasma cholesteryl ester transfer protein (CETP) activity were examined in 12 healthy young women. Subjects followed the two experimental diets for 4 weeks according to a randomized crossover design. Both experimental diet periods were preceded by consumption of a baseline diet for 2 weeks. The diets provided 37% of total energy intake (E%) as fat, and differed only with respect to fatty acid composition. There was a substitution of 5E% of palmitic acid or stearic acid in the experimental diets for 5E% of monounsaturated fatty acids in the baseline diet. After the palmitic acid diet, serum total and high-density lipoprotein (HDL) cholesterol and apo A-I concentrations were higher (8%, P = .015, 9%, P = .040, and 11%,P = .011, respectively) and mean serum low-density lipoprotein (LDL) cholesterol concentration tended to be higher (8%, P = .077) as compared with values after the stearic acid diet. Plasma CETP activity increased in the palmitic acid diet as compared with the stearic acid diet (12%, P = .006). In conclusion, palmitic acid and stearic acid-enriched diets had different effects on serum lipids and lipoproteins and also on plasma CETP activity in young healthy women."

 

 

Strikes me that identifying foods which have a balance towards stearic acid and away from palmitic acid is a good idea.

 

Edited by johnhemming, 03 October 2022 - 09:53 AM.

[Learner056]

I do am afraid of Palmitic because of my mutations, I get canker sores, and learnt this travesty while back that USP is an "enabler" of this dog-whistle on letting Stearic be 50% Palmitic.  Over the years I have countered my Palmitic side-effects with Kidney extract that like magic heals canker sore (it helps with histamine issues too).  I am bit doubtful though if indeed Palmitic is causing this, but maybe it is, thank you for explanation, will now see.  

 

It is quite possible that stearic acid is not the culprit for hypertension, but palmitic acid. 

 

 

 

 

John, you talked about HIF-1a earlier I think.  I am now trying to understand the "fuel systems" cross-talk in Stem cells.  I believe it is a mistake to view stem cell population as one monolithic entity. I have reason to believe.  When you improve one organ, you unknowingly deteriorate a different organ. 

 

a) Stem cells (various types) use various fuel sources.  Have you (or anyone else here) studied this area and if so what is the distribution like? which stem cell uses which fuel.  This is critical. 

b) I have observed that Lactate is potent in certain types of stem cells.  HIF-1a is related to Lactate I think.  I tried NAD+ on my face (may it be amazing molecule for the body) but it made me look 10 yrs older if not more.  I lost all my subcutaneous fat leaving behind wrinkles and ravage.  I am now trying to undo that damage with Lactic acid, and it seems to be a wonder stimulant for epithelial tissue i.e. face, so far, jury is out, so I should not be presumptuous (as you young like to say)

 

 

 

Edited by Learner056, 03 October 2022 - 09:49 PM.

[johnhemming]

John, you talked about HIF-1a earlier I think.  I am now trying to understand the "fuel systems" cross-talk in Stem cells.  I believe it is a mistake to view stem cell population as one monolithic entity. I have reason to believe.  When you improve one organ, you unknowingly deteriorate a different organ. 

 

 

 

HIF 1 alpha is one of the systems that respond to hypoxia the other two being NRF2 and NF kappa B.

 

HIF 1 alpha is generally a helpful response.

 

If you do the right things you should be able to improve all organs, but if you interfere with homeostasis wrongly it can have negative effects.  Hence for example Pantethine can increase bilirubin through extending bleeding. (probably because it is not rate limited and generates too much of something probably coenzyme A) 

[Learner056]

About PQQ: "A decrease in mitochondrial potential, the promotion of mitochondrial fusion and the accelerated movement of mitochondria were all observed in PQQ-pretreated cells" Ref: https://www.nature.c...514-022-00083-0

Why is decrease in mitochondrial potential good?  (I thought larger mitochondrial membrane potential was good, from other readings, larger potential equates larger ATP). 

[Learner056]

Anyone care to explain?  thank you. 

 

About PQQ: "A decrease in mitochondrial potential, the promotion of mitochondrial fusion and the accelerated movement of mitochondria were all observed in PQQ-pretreated cells" Ref: https://www.nature.c...514-022-00083-0

Why is decrease in mitochondrial potential good?  (I thought larger mitochondrial membrane potential was good, from other readings, larger potential equates larger ATP). 

 

[Learner056]

People no longer care, our society is in decline.  

 

About PQQ: "A decrease in mitochondrial potential, the promotion of mitochondrial fusion and the accelerated movement of mitochondria were all observed in PQQ-pretreated cells" Ref: https://www.nature.c...514-022-00083-0

Why is decrease in mitochondrial potential good?  (I thought larger mitochondrial membrane potential was good, from other readings, larger potential equates larger ATP). 

 

 

Anyone care to explain?  thank you. 

 

[Learner056]

For the record, my criticism (if you call it such) is not directed towards Turnbuckle at all.  It is directed towards the "collective us" i.e. you the biohacker community of thousands here, who should be knowing this all, and sharing it, and helping to explain to others.  Indeed, Turnbuckle is #1 brilliant mind of this century - I am going to attest to that (even though I may not agree with him fully), but this protocol (with certain refinements) is indeed a piece of magnificent art stitched with the real science (vs many misleading material out there) for times to come.  The fact that he shared with all, oh boy I know, many would not be happy with him (a big understatement).  But he did still, that is courage, and that is ethics of humanity.  But he is 1 person.  But "you" are many.  Peace. 

 

Edited by Learner056, 12 October 2022 - 05:57 PM.

[FWP]

Could taurine be of use to the protocol? The nicotinamide can have a negative effect on mood for me and taurine helps. Or taurine is undoing this part of the protocol? It does seem to have an effect on the mitochondria. https://www.scienced...213231721000690

[Turnbuckle]

Could taurine be of use to the protocol? The nicotinamide can have a negative effect on mood for me and taurine helps. Or taurine is undoing this part of the protocol? It does seem to have an effect on the mitochondria. https://www.scienced...213231721000690

 

The point is to isolate, detect, and remove defective mtDNA. If you aid dysfunctional mitochondria, you may undo what you are trying to achieve. However, feel free to try it and see if the curls to failure improve during fission.

Edited by Turnbuckle, 26 October 2022 - 08:19 PM.

[ortcloud]

new fission candidate?

 

New study suggests that vitamin K2 promotes mitochondrial fission by activating the PINK1/Parkin signaling pathway thereby regulating the mitochondrial quality-control loop

 

 

https://www.ncbi.nlm...les/PMC9003256/

 

Turnbuckle, should we throw this into the stack?

[EliotH]

Crap. I take Koncentrated K every day which has 25mg MK-4 and .5 mg MK-7. That might cause a problem on fusion days.

[Repack Racing]

Yes - take K/D supps on fission day.

[EliotH]

I've just embarked on the protocol having modified rarefried's version as follows:  First day AKG + PQQ + 150 mg (non-liposomal) apigenin + 350 mg of niacinamide for fission; second day AKG + PQQ + 350 mg niacinamide; and on the third AKG + PQQ + 3 g GMS + 800 mg DHM. 

 

I'm curious whether "less is more" holds true for niacinamide, and I don't think it will hurt to toss the riboside.   

 

(I just saw this post.) I think Ray Peat and his followers believe niacinamide is better at low doses, like 250 mg divided into 2 or 3 doses per day IIRC (I don't feel like looking it up at the moment).

[Empiricus]

(I just saw this post.) I think Ray Peat and his followers believe niacinamide is better at low doses, like 250 mg divided into 2 or 3 doses per day IIRC (I don't feel like looking it up at the moment).

 

Interesting idea.  The main objection to taking smaller doses throughout the day is the likelihood that chronic dosing could damage the liver.  Time release niacin seemed to do that.  However, since those of us following this Turnbuckle protocol would not be intermittently dosing niacinamide throughout every single day--as some Ray Peat followers seem to be doing--this caveat might not apply so much to us.

 

Nevertheless, as I recall from the early days of this protocol, seeking to avoid this risk altogether was actually part of the reason Turnbuckle chose not divide niacinamide into spread-out smaller doses.  

Edited by Empiricus, 29 November 2022 - 11:02 AM.

[EliotH]

Looks like Turnbuckle left the site completely on 11/26. He was getting a lot of shit posts on the Stem Cell thread and the idiots wouldn't stop despite warnings. LongeCity doesn't have any way for thread starters to delete posts or ban problem posters. Too bad, our loss.

 

ETA: Looks like he deleted his protocols too.

Edited by EliotH, 01 December 2022 - 03:09 PM.

[Empiricus]

ETA: Looks like he deleted his protocols too.

 

Turnbuckle only deleted the links. The protocols remain in their respective threads. 

[EliotH]

Turnbuckle only deleted the links. The protocols remain in their respective threads. 

 

He used to have his latest versions on his profile page, now you would nee to dig through the threads backwards to find the latest. Fortunately I saved them off to a local text file. Except the SC one, I don't recall if I got the latest version.

[Repack Racing]

What happened on the C60 thread was absolutely ridiculous.  We just need to ignore these people who are bent on derailing things. 

 

Here is a link to the latest protocol: https://www.longecit...-58#entry903440

 

Note that TB added some supps to the C60 fusion and fission days.  He was planning to update the mito protocol before things went south.

 

Here is a link to the latest C60 protocol: https://www.longecit...-66#entry917974

 

 

An updated Mito protocol

 

The previous protocol can be found at post #1366

The new protocol:

 

This new procedure is much simplified. It requires only two doses, Mito1 and Mito2, which are alternated on a daily basis.

 

Mito1 (fission)

● NAM+R, 1 g of each

● AKG, 1 g

● PQQ, 20 mg

 

Mito2 (fusion)

● GMS, 1 g

● AKG, 1 g

● PQQ, 20 mg

 

 

Edited by ildr, 04 December 2022 - 12:47 AM.

[Repack Racing]

Ok community, after many years of study and experimentation with this and other protocols, I have developed my new protocol, which I will run for 6 weeks and then do an age test. The principle is Fission => Autophagy => Fusion + stem cells and senescence.

 

A few notes: I realize this is a complicated protocol in contrast to TB's efforts to simplify.  However, it does draw from TB and many others here, as well as in the Stem Cell/C60 thread.  Secondly, it is impractical to provide references for all the items in this protocol.  I am happy to provide specific references on request.  I have taken TB's approach of multiple pathways/actions for each phase - Fission / Autophagy / Fusion. The F-A-F cycle is continuous and can be very short.  The concept here is simply to enhance one over the other.

 

I am relying on the low-end 16 hour fast for autophagy.  I realize this may be controversial.  I am an athlete and also want to enjoy my (hopefully long) life and just can't do full 24h+ fasting.  I have added autophagy promoters to help. It is not ideal, but  we will see.

 

If anyone sees any flaws or red flags in this protocol, please speak up.

 

The protocol:

 

7-Day Fission/Autophagy/Fusion cycle

 

Day 1 – Fission / Senescence

 

Regular Diet/Calories

 

Wake-up – Fission:

• NMN – 1g
• Lipo NR – 600 mg
• Core Lipo B-complex – 3 caps
• Nicotinamide – 1 g
• Lipo CaAKG – 300 mg
• CaAKG – 1 g

30 Minutes – Other:

• ZMA
• Multi
• CoQA

1 Hour – Senescence:

• Lipo Fisetin – 3 g
• Quercetin – 600 mg
• Apigenin – 1 g
• Grape Seed Extract – 2 g
• Bioprene – 20 mg
• Pterostilbene – 250 mg

7 Hours

• NACET/GlyNAC (stops fusion/autophagy/excess fission) 600 mg / 600 mg

Notes:

• Avoid autophagy supplements/fasting during day.

Day 2 – Autophagy/Mitophagy

 

Start fast (7PM or so the night before)

Electrolytes (no calories)

 

Wake up – Autophagy Promoters:

• NMN – 1 g
• Lipo Spermidine – 8 mg
• Acetyl Glucosamine (NAG) – 700 mg
• Pomegranate – 400 mg
• Autophagy Renew Luteolin/Piperlongumine 165 mg / 10 mg
• PQQ – 20 mg
• Electrolytes (no calories)

Plus:

• ZMA
• CoQA

1-2 hours vigorous exercise

 

18 Hours

• C60 (induces autophagy but stops fast)

19 Hours

• Resume regular diet/calories

Notes:

• No antioxidants during autophagy
• No amino acids during autophagy
• No Senolytics during autophagy

Day 3 – Fusion

 

Wake-up:

• Mots-C - 5 mg
• GMS – 3 g
• DHM – 3 g
• PQQ – 20 mg
• Lipo CaAKG – 600 mg
• CaAKG – 1 g
• Liposomal Vitamin D3K2 – 2,500 iu / 200 mcg
• Beta Glucan – 50 mg

30 Minutes – Other:

• ZMA
• Multi
• CoQA
• Lysine, Threonine, Methionine if needed

1-2 hours vigorous exercise

 

5-7 Hours

• C60
• Lysine, Threonine, Methionine if needed

10 Hours

• NACET/GlyNAC (stops fusion)

Day 4 – Fission

 

Wake-up – Fission:

• Same as Day 1

No Senolytics

 

30 Minutes – Other:

• Same as Day 1

7 Hours

• Same as Day 1

Day 5 – Autophagy/Mitophagy

 

Start fast (7PM or so the night before)

Electrolytes (no calories)

 

Wake up – Autophagy Promoters:

• Same as Day 2

1-2 hours vigorous exercise

 

18 Hours

• C60 (induces autophagy but stops fast)

19 Hours

• Resume regular diet/calories

Day 6 – Fusion/Stem Cells

 

Wake-up:

• Stearic acid chocolate bar

3 Hours:

• Same as Day 3
• Add: C60
• Delete: Mots-c

3.5 Hours – Other:

• Same as Day 3

1-2 hours vigorous exercise

 

5-7 Hours

• Same as Day 3

Day 7 – Rest/Light Autophagy (for stem cells)

 

Wake up – Autophagy Promoters:

• Same as Day 2/5

Fast until 12PM.

 

10 Hours:

• NACET/GlyNAC

 

Repeat cycle

 

May add: TMG for methyl stores.

 

Edited by ildr, 05 December 2022 - 08:23 PM.

[Female Scientist]

Thanks for your hard work on this. I would very much like to see a "lite" version that non-fasters could use. I am not able to fast much past say, 14 hours overnight, (nor am I able to perform intense exercise) due to migraine and blood sugar issues. Thankfully these both are improving due to following versions of TB's protocols over the years, but I would ideally like a "lite" version of this protocol which would suit my situation. Just throwing it out there in case anyone has recommendations.

Ok community, after many years of study and experimentation with this and other protocols, I have developed my new protocol, which I will run for 6 weeks and then do an age test. The principle is Fission => Autophagy => Fusion + stem cells and senescence.

 

A few notes: I realize this is a complicated protocol in contrast to TB's efforts to simplify.  However, it does draw from TB and many others here, as well as in the Stem Cell/C60 thread.  Secondly, it is impractical to provide references for all the items in this protocol.  I am happy to provide specific references on request.  I have taken TB's approach of multiple pathways/actions for each phase - Fission / Autophagy / Fusion. The F-A-F cycle is continuous and can be very short.  The concept here is simply to enhance one over the other.

 

I am relying on the low-end 16 hour fast for autophagy.  I realize this may be controversial.  I am an athlete and also want to enjoy my (hopefully long) life and just can't do full 24h+ fasting.  I have added autophagy promoters to help. It is not ideal, but  we will see.

 

If anyone sees any flaws or red flags in this protocol, please speak up.

 

The protocol:

 

7-Day Fission/Autophagy/Fusion cycle

 

Day 1 – Fission / Senescence

 

Regular Diet/Calories

 

Wake-up – Fission:

• NMN – 1g
• Lipo NR – 600 mg
• Core Lipo B-complex – 3 caps
• Nicotinamide – 1 g
• Lipo CaAKG – 300 mg
• CaAKG – 1 g

30 Minutes – Other:

• ZMA
• Multi
• CoQA

 

[Repack Racing]

Thanks for your hard work on this. I would very much like to see a "lite" version that non-fasters could use. I am not able to fast much past say, 14 hours overnight, (nor am I able to perform intense exercise) due to migraine and blood sugar issues. Thankfully these both are improving due to following versions of TB's protocols over the years, but I would ideally like a "lite" version of this protocol which would suit my situation. Just throwing it out there in case anyone has recommendations.

 

Female Scientist,

 

You can absolutely simplify this.  I'd continue with Turnbuckle's protocol above.  He claims an epigenetic age of -20.

 

If you want to add Stem Cell proliferation, sprinkle in the protocol from that thread on fission day every once in a while (link above).

 

The most common senolytics are fisetin and apigenin.  Grape seed extract is good too. For the senolytic effect, the doses must be large.  However, it's good idea to start with less to see how you react.

 

Try to get liposomal versions if possible.

 

With blood sugar issues, I wouldn't worry too much about fasting.  Perhaps stop eating at 6PM and then resume at 9AM or so - that's 15 hrs.  You can try the autophagy supplements, but reviews are mixed.

 

NACET and GlyNAC are powerful antioxidants that help balance the mitochondria and help ameliorate excess fission and fusion.  If you add, make sure to space it out from the other parts of the protocol.

 

Good luck!

 

Edited by ildr, 06 December 2022 - 01:42 AM.

[stephen_b]

ildr, are you expecting results from this showing up on a trume-like test? I know TB had the dumbell test for this protocol and the epigenetic age test for the stem cell protocol.

[Repack Racing]

ildr, are you expecting results from this showing up on a trume-like test? I know TB had the dumbell test for this protocol and the epigenetic age test for the stem cell protocol.

 

stephen_b,

 

I am a competitive athlete, so I am not sure the dumbell test would be particularly helpful for me.  I can definitely tell the general track of my performance, which will be an indicator.

 

I purchased the ProHealth age test, which no one here has done that I know of. I chose it due to ProHealth's supplements consistently coming back with the highest purity in third-party supplement testing. I figure if they can nail that then the test will be good.

Edited by ildr, 06 December 2022 - 02:31 AM.

[Heisok]

Good information.

 

ildr, would you create your own thread?

 

Thanks.

[Empiricus]

Good information.

 

ildr, would you create your own thread?

 

Thanks.

 

idlr, I agree with Heisok. What you're attempting here is sufficiently ambitious that it deserves its own thread. The mito protocol is not intended to be anti-aging, it's mainly for restoring mito function. Hence Stephan's question. 

 

One of the main reasons Turnbuckle initially separated the two protocols was to create a mito protocol that young people can safely use.  Turnbuckle discourages young people from taking c60--even with fusion supps.   Turnbuckle and others have demonstrated that mito protocol works fine without c60.  Even in offering you feedback for your protocol, I would be derailing this thread with thoughts about c60 that don't belong here.  

 

Of course, by all means drop links here or over at the other thread as you deem them relevant.   

[smithx]

Trying to do everything in one day is too much and is very likely to not work (I'm referring specifically to the Day 1 protocol including fission and senolytics spaced by an hour).

 

It takes a while for these compounds to get through the body, and you can't rush things, or you end up with them all being loaded at once and potentially canceling each other out or causing other issues (for example kidney or liver overloading).

 

Pharmacokinetics are important to take into account.

 

My method is to to do senolytics every 8 days, two days before my rapamycin dose, for the above reasons (and others). 

The only thing I do daily (otherwise) is anti-HERV supplements because HERV needs to be suppressed all the time.

 

 

Edited by smithx, 07 December 2022 - 07:06 AM.