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Manipulating mitochondrial dynamics

nad nad+ c60 mito fission fusion stearic acid mtdna methylene blue

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#961 pone11

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Posted 12 June 2018 - 07:47 PM

Are there any studies showing lifespan extension with mitochondrial fission?

 

How do we know the appropriate amounts of time to remain in fission versus fusion states?  Other than totally subjective reports of how we feel, is there any objectively measured outcome to show that this protocol accomplished its objectives?


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#962 pone11

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Posted 12 June 2018 - 09:10 PM

So after a brief review of mitochondrial dynamics of fusion and fission, here are some thoughts:

 

* Reviewing fusion and fission, what is very very clear is that fusion is absolutely critical to being energetic and healthy.  When the key proteins involved in fusion fail, you get a number of very serious diseases, some of which are profiled in the summary article above.   The proper function of fusion is critical to peak bioenergetics of the cell, so if you want to oxidize aerobically and efficiently, you must have good fusion.

 

* Fission is more of a cloud.   There is a clear association with autophagy and mitophagy and reduced bioenergetics.   One would assume that this mitochondrial state helps to support fasting or starvation, by removing non-optimal structures from the cell.   The problem is that the studies showing lifespan extension from autophagy are murky.   And I have found at least one study in worms where suppressing autophagy as the worms age EXTENDS LIFESPAN.

 

I am not saying that it plays out in humans the way it plays out in worms.   But I am saying that the science behind autophagy and mitochondrial fission really is not at a level where you can definitively say that fission is good for lifespan extension.   By hypothesis only, maybe it ends up being shown that autophagy is a necessary adaptation for a low-energy environment, but that - on net - the absence of high bioenergetics to the cell is far more harmful than the presence of fission during autophagy / mitophagy is good.   There is a complex system at work here and we are making a simplistic assumption that because mitophagy does something favorable that entering into the state where it occurs is on net beneficial to our bodies.  Maybe yes.  Maybe no.   The worm study I linked above should make you think twice.

 

This then feeds into my question posted earlier:  even if you want to believe that fission is good for lifespan extension, how much is enough?  How much is actually harmful?   How long do these cycles need to last to be therapeutic?   Do we have studies answering any of those questions in any kind of organism?

 


Edited by pone11, 12 June 2018 - 09:12 PM.

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#963 Turnbuckle

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Posted 12 June 2018 - 10:15 PM

Mitochondria are not static. They are constantly merging and dividing, and mito morphology is used by cells for certain processes. The health of mitochondria is one of them. Fission is required for mitophagy, and without it bad loops of mtDNA will not be removed. They will build up unnoticed until it's too late. The fused state is more efficient at producing ATP as it disperses mtDNA loops and mito components, thereby diluting the effects of bad components, particularly bad loops. But because of this averaging effect in the fused state, cells cannot detect bad mitochondria (nor can lysosomes digest them).

 

If the natural process of fission/fusion were sufficient for quality control, mitochondria would not be a source of aging. But mito QC falls behind over the decades and bad mitochondria build up. Using this protocol drives mitochondria all the way in one direction and then the other, eliminating bad mitochondria and replacing them via biogenesis, thus clearing out the bad population in an iterative fashion. You will know that you've done enough of it when the fission state doesn't seem much different than the fusion state.

 

Mito morphology also directs the fate of stem cells during proliferation. Fission biases stem cells to asymmetric division while fusion biases it to symmetric division, also called self-renewal. See the thread Stem cell self-renewal with C60, where I argue that using C60 in a fission state will deplete the stem cell pool, while using it in the fusion state will enlarge the pool.

 

Note: As the fission/fusion and stem cell protocols are my ideas, you will not find any animal research on them.


Edited by Turnbuckle, 12 June 2018 - 10:52 PM.

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#964 sthira

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Posted 12 June 2018 - 10:55 PM

...If the natural process of fission/fusion were sufficient...

General question asked in the spirit friendly curiosity and not in criticism of anything you're trying: are you seeking substance combinations that may mimic the overall states of prolonged fasting and refeeding?

Hope I'm not off topic, but the title says "Manipulating mitochondrial dynamics," and prolonged fasting seems to tear pieces down; healthy refeeding seems to rebuild them. But what remains internally aging right along is the organism itself -- each cell is living and dying at its own unique pace. Even if we do manage to eliminate zombie mitochondria, rebuild them, and kill off senescent cells, the organism is still aging anyway. Having ways to measure if anything is happening by using our various protocols would be nice, but it still wouldn't be rejuvenation.

Slow down aging damage, stop it, then reverse the damages -- these seem like three separate humongous tasks.

Edited by sthira, 12 June 2018 - 11:02 PM.

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#965 Turnbuckle

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Posted 12 June 2018 - 11:14 PM

(1) General question asked in the spirit friendly curiosity and not in criticism of anything you're trying: are you seeking substance combinations that may mimic the overall states of prolonged fasting and refeeding?

(2) Hope I'm not off topic, but the title says "Manipulating mitochondrial dynamics," and prolonged fasting seems to tear pieces down; healthy refeeding seems to rebuild them. But what remains internally aging right along is the organism itself -- each cell is living and dying at its own unique pace. Even if we do manage to eliminate zombie mitochondria, rebuild them, and kill off senescent cells, the organism is still aging anyway. Having ways to measure if anything is happening by using our various protocols would be nice, but it still wouldn't be rejuvenation.

Slow down aging damage, stop it, then reverse the damages -- these seem like three separate humongous tasks.

 

1. No, I wasn't trying to do that. I came at this directly from the research that showed large amounts of nicotinamide eliminated mitochondria. And that a high NAD+/NADH ratio fissioned them, making them subject to mito QC. 

 

2. Fission/fusion deals with bad mitochondria as a source of aging, but there are other sources. A number of them. Depletion of stem cells is a big one, and my stem cell protocol is directed to that. Epigenetic and telomeric aging are two more (sometimes referred to as clocks), and while ways of dealing with these have been explored, reducing one seems to accelerate the other. In my thread Reversing the Clocks, I suggested a method of reversing both.


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#966 pone11

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Posted 12 June 2018 - 11:24 PM

Mitochondria are not static. They are constantly merging and dividing, and mito morphology is used by cells for certain processes. The health of mitochondria is one of them. Fission is required for mitophagy, and without it bad loops of mtDNA will not be removed. They will build up unnoticed until it's too late. The fused state is more efficient at producing ATP as it disperses mtDNA loops and mito components, thereby diluting the effects of bad components, particularly bad loops. But because of this averaging effect in the fused state, cells cannot detect bad mitochondria (nor can lysosomes digest them).

 

If the natural process of fission/fusion were sufficient for quality control, mitochondria would not be a source of aging. But mito QC falls behind over the decades and bad mitochondria build up. Using this protocol drives mitochondria all the way in one direction and then the other, eliminating bad mitochondria and replacing them via biogenesis, thus clearing out the bad population in an iterative fashion. You will know that you've done enough of it when the fission state doesn't seem much different than the fusion state.

 

Mito morphology also directs the fate of stem cells during proliferation. Fission biases stem cells to asymmetric division while fusion biases it to symmetric division, also called self-renewal. See the thread Stem cell self-renewal with C60, where I argue that using C60 in a fission state will deplete the stem cell pool, while using it in the fusion state will enlarge the pool.

 

Note: As the fission/fusion and stem cell protocols are my ideas, you will not find any animal research on them.

 

The theory is good, yet when you test a counter-theory by suppressing autophagy in worms, the worms live longer.   

 

I want to believe that mitophagy is both necessary and healthful and that doing it on a regular basis is a good health practice.  But the experimental evidence is very underwhelming.   For every study like this one that claims autophagy promotes lifespan, you get a study like this one where suppressing autophagy extends lifespan.   There is no clear trend, and there are not many studies.

 

Compare this against the thousand+ studies being done on sulforaphane and urolithin (from pomegranate seeds) which have shown overwhelming benefits and duplicate over and over and the story actually just keeps getting better and stronger.   Those are sure things, at least much as anything can be that is based on ongoing research.   By comparison, mitophagy and autophagy protocols are speculative and not confirmed by research as healthful practices.


Edited by pone11, 12 June 2018 - 11:25 PM.

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#967 Turnbuckle

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Posted 12 June 2018 - 11:51 PM

Mitophagy is a form of autophagy, but they are not the same thing. As for these protocols being speculative, they are and I don't claim otherwise. I have used them and gotten benefit from them, and I have put these up for the benefit of others. If you want to wait for animal trials, you are not alone, however many people on this forum are into self-experimentation and aren't interested in waiting for animal trials that may never come, at least in their lifetimes.


Edited by Turnbuckle, 12 June 2018 - 11:57 PM.

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#968 pone11

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Posted 13 June 2018 - 01:03 AM

Mitophagy is a form of autophagy, but they are not the same thing. As for these protocols being speculative, they are and I don't claim otherwise. I have used them and gotten benefit from them, and I have put these up for the benefit of others. If you want to wait for animal trials, you are not alone, however many people on this forum are into self-experimentation and aren't interested in waiting for animal trials that may never come, at least in their lifetimes.

 

It's all reasonable hypothesis, and yes this site is for those who do not mind being on a the leading edge.

 

What are your thoughts on the minimum time that a person should spend in mitophagy each week?   In your view, would it be enough to push fusion six days a week and dedicate one day to fission?

 

Trying to regulate these processes over the course of a single day seems really speculative.  It takes the body time to ramp up and down processes and the chances of getting everything synchronized seems hit or miss.   But maybe dedicating an entire day to the autophagy/mitophagy idea might at least make for a decent experiment.


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#969 Turnbuckle

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Posted 13 June 2018 - 01:47 AM

The fission/fusion thread is very long and I tried a number of versions of the protocol along the way, so I posted a link to one on my profile page, and also to the other protocols.


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#970 pone11

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Posted 13 June 2018 - 02:50 AM

The fission/fusion thread is very long and I tried a number of versions of the protocol along the way, so I posted a link to one on my profile page, and also to the other protocols.

 

I guess the question was not so much what is your protocol, but more why do you believe that a minimum of X hours / days of mitochondrial fission per week is optimal.



#971 Turnbuckle

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Posted 13 June 2018 - 09:59 AM

I guess the question was not so much what is your protocol, but more why do you believe that a minimum of X hours / days of mitochondrial fission per week is optimal.

 

 

That discussion goes back a couple of years to a previous thread -- A protocol to upgrade mitochondria. In the first post, see the first figure in the first reference -- Nicotinamide-induced Mitophagy. 

 

Fig. 1A shows an example of the change in the mitochondrial content that is commonly observed in various tested human cells (including normal fibroblasts and MCF-7, H460, and HCT116 cancer cell lines) after supplementation of 5 mM NAM in culture media. The mitochondrial content as determined by flow cytometry using two different mitochondrion-specific dyes was substantially decreased for the first 3 days and thereafter remained at a level about 70% that in the untreated cells, as reported previously (23).

http://www.jbc.org/c...7/23/19304.long

 

 


Edited by Turnbuckle, 13 June 2018 - 10:56 AM.

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#972 pone11

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Posted 13 June 2018 - 11:00 PM

That discussion goes back a couple of years to a previous thread -- A protocol to upgrade mitochondria. In the first post, see the first figure in the first reference -- Nicotinamide-induced Mitophagy. 

 

If you have not already considered it, look at urolithin as a mitophagy inducer.  This metabolite is derived from tannins in pomegranate seeds and there is amazingly positive research behind it.   Be careful taking this from supplements as the vast majority do not have the right tannins in them.   I can share research on that if you decide to look further.



#973 APBT

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Posted 14 June 2018 - 12:53 PM

If you have not already considered it, look at urolithin as a mitophagy inducer.  This metabolite is derived from tannins in pomegranate seeds and there is amazingly positive research behind it.   Be careful taking this from supplements as the vast majority do not have the right tannins in them.   I can share research on that if you decide to look further.

 

See this thread:  https://www.longecit...y-fight-ageing/



#974 Turnbuckle

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Posted 14 June 2018 - 04:55 PM

If you have not already considered it, look at urolithin as a mitophagy inducer.  This metabolite is derived from tannins in pomegranate seeds and there is amazingly positive research behind it.   Be careful taking this from supplements as the vast majority do not have the right tannins in them.   I can share research on that if you decide to look further.

 

 

This was considered on the second page of the ancestor thread to this one--A protocol to upgrade mitochondria. I'm not sure if anyone tried it as N+R seemed sufficiently effective.


Edited by Turnbuckle, 14 June 2018 - 04:56 PM.

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#975 onz

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Posted 16 June 2018 - 12:14 AM

I have read through most of this huge thread, but still have a couple of questions.

 

What is a suitable alternative to stearic acid? I cannot find a Canadian distributor for food-grade stearic acid and the shipping charges for other countries are ridiculous. 

 

What are you all using for your source of hydroxytyrosol? The most pure extract I could find was by ProHealth, but again the shipping cost to Canada is outrageous.



#976 Turnbuckle

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Posted 16 June 2018 - 09:39 AM

I have read through most of this huge thread, but still have a couple of questions.

 

What is a suitable alternative to stearic acid? I cannot find a Canadian distributor for food-grade stearic acid and the shipping charges for other countries are ridiculous. 

 

What are you all using for your source of hydroxytyrosol? The most pure extract I could find was by ProHealth, but again the shipping cost to Canada is outrageous.

 

 

Stearic acid is by far the best that I know of. If you can't find food grade, look for NF/USP grade -- "A chemical grade of sufficient purity to meet or exceed requirements of the United States Pharmacopeia (USP); acceptable for food, drug, or medicinal use; may be used for most laboratory purposes."

 

This appears to be available on Amazon/Canada at the moment, though 4 times more expensive than in the US.

 

If you can't find hydroxytyrosol, leave it off.


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#977 Turnbuckle

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Posted 16 June 2018 - 09:48 AM

Five-day protocol, updated

 

 

Day 1-2: Fission/mitophagy

 

Morning

Nicotinamide (NAD+) — 2g

Ribose (NAD+) — 2g

Jiaogulan leaf (AMPK activator) — 1g

Apigenin (fission) — 100mg

Fisetin (Sirt1 activator) — 100mg

 

Evening

Jiaogulan leaf — 500mg

Fisetin — 100 mg

 

Day 3: Fission/mitophagy

 

Morning & Evening

Jiaogulan leaf  — 500mg

Fisetin — 100 mg

 

Day 4: Fusion/biogenesis

 

Morning

Stearic acid (fusion) — 5-10 g

Leucine (biogenesis) — 5g

PQQ (biogenesis) — 20mg

Hydroxytyrosol (biogenesis & antioxidant) — 25mg

Vitamin B complex (commercial mix)

 

Evening

Leucine — 5g

PQQ — 20mg

Hydroxytyrosol — 25mg

 

Day 5: Fusion/biogenesis

 

Morning & evening

Leucine  — 5g

PQQ  — 20mg

Hydroxytyrosol  — 25mg

 

 

Notes:

Stearic acid — use either “food grade” or NF/USP grade.

Hydroxytyrosol — if unavailable where you live, leave it off.

 

 


Edited by Turnbuckle, 16 June 2018 - 10:02 AM.

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#978 aribadabar

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Posted 16 June 2018 - 01:29 PM

 I cannot find a Canadian distributor for food-grade stearic acid and the shipping charges for other countries are ridiculous. 

 

What are you all using for your source of hydroxytyrosol? The most pure extract I could find was by ProHealth, but again the shipping cost to Canada is outrageous.

 

Ship to US side of the border and go pick it up in a day trip?



#979 adamh

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Posted 16 June 2018 - 07:09 PM

I'm not seeing much about the role of deep red and near infrared light in this thread. It clearly affects mito dynamics. I've been using it myself with great results. Its excellent for osteo, aches and pains, many other things too. It reenergizes the mitochondria you already have and makes them work better. Apparently they absorb certain wavelengths of light much as plant cells do and use it.

 

Light may also affect fusion and fission as part of the process. It seems very important and unlike with drugs you do not get side effects. Here are some interesting links with proposed explanations of the mechanism

 

https://www.newscien...ging-our-cells/

 

http://www.warp-ligh...sWongWhelan.pdf


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#980 Turnbuckle

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Posted 16 June 2018 - 07:27 PM

I'm not seeing much about the role of deep red and near infrared light in this thread. It clearly affects mito dynamics. I've been using it myself with great results. Its excellent for osteo, aches and pains, many other things too. It reenergizes the mitochondria you already have and makes them work better. Apparently they absorb certain wavelengths of light much as plant cells do and use it.

 

Light may also affect fusion and fission as part of the process. It seems very important and unlike with drugs you do not get side effects. Here are some interesting links with proposed explanations of the mechanism

 

https://www.newscien...ging-our-cells/

 

http://www.warp-ligh...sWongWhelan.pdf

 

It's also known to stimulate stem cell proliferation, and that process is set in motion by mitochondria beginning to produce ATP. A few days ago I started a thread called Losing weight at light speed, in which I used red light and fasting to achieve a previously unattainable (for me) rate of weight loss. So you might want to post these links over there as well, as some think it must be a placebo effect.


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#981 stephen_b

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Posted 16 June 2018 - 08:01 PM

PMID 24513856: A small natural molecule promotes mitochondrial fusion through inhibition of the deubiquitinase USP30.

 

 

  1. Here, we identified a diterpenoid derivative 15-oxospiramilactone (S3) that potently induced mitochondrial fusion to restore the mitochondrial network and oxidative respiration in cells that are deficient in either Mfn1 or Mfn2.

 

(Emphasis mine.)

 

This is referred to as a 'small natural molecule', but I was not able to find whether there is a natural source for it. Might be a nice tool to have.



#982 Turnbuckle

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Posted 16 June 2018 - 08:10 PM

Sulforaphane works as well, supposedly, but I've never liked the feel of it.


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#983 onz

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Posted 18 June 2018 - 07:56 AM

Would using one of the Olive Leaf Extracts available on iHerb, which are standardising to Oleuropein content, be ok? Or would some of the additional compounds interfere with the fusion goals?



#984 Turnbuckle

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Posted 18 June 2018 - 10:13 AM

Would using one of the Olive Leaf Extracts available on iHerb, which are standardising to Oleuropein content, be ok? Or would some of the additional compounds interfere with the fusion goals?

 

It likely has little hydroxytyrosol, but I wouldn't expect the oleuropein to hurt anything, as it is also known to support biogenesis. As for unknown components, I can't say.


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#985 HaplogroupW

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Posted 21 June 2018 - 05:46 PM

Consider adding L-carnitine and/or acetyl-l-carnitine to the fission protocol for better mitophagy:

 

 

https://www.scienced...026049517302494

 

This is the first study to suggest that the activation of autophagy may be a novel mechanism by which carnitine treatment enhances mitochondrial function. After six weeks of a high-fat diet, the mitochondria in the skeletal muscle of mice were swollen with destroyed cristae and a reduction in mitochondrial function. Treatment with carnitine for two weeks eliminated the malfunctioning mitochondria and resulted in an increase in the formation of autophagosomes with a concomitant increase in PPARγ. In a cellular model of free fatty acid (palmitate)-induced insulin resistance, we observed the repression of autophagy, which was reversed by pre-incubation with carnitine in a dose-dependent manner. However, autophagy was not activated by carnitine after PPARγ was silenced, suggesting that autophagy activation by carnitine occurs through PPARγ.

 

 

 

When abnormal mitochondria that can no longer be repaired by physiological fission and fusion machinery accumulate, mitochondrial autophagy to remove the damaged mitochondria should be activated to maintain cellular health. The defective turnover of mitochondria in the pathogenesis of type 2 diabetes has been suggested [21]. In the present study, autophagy was suppressed in high-fat diet-fed mice and palmitate-treated myotubes, and mitochondria with abnormal morphology predominated in the skeletal muscle of high-fat diet-fed mice. Carnitine treatment in insulin resistance activated autophagy and improved both mitochondrial function and insulin resistance.

 

 

 

To summarize, we observed the clearance of abnormal mitochondria by mitochondrial autophagy after carnitine treatment in an insulin-resistant rodent model and suggest that carnitine activates autophagy through PPARγ in a cellular model of free fatty acid-induced insulin resistance. Further investigation is required to determine if other factors are involved in the regulation of autophagy by carnitine.

 


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#986 BieraK

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Posted 21 June 2018 - 09:46 PM

I wonder if N+R in low doses boost NAD+ comparable to NR, and if low doses NR promotes fission as high dose



#987 aribadabar

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Posted 22 June 2018 - 04:04 PM

Consider adding L-carnitine and/or acetyl-l-carnitine to the fission protocol for better mitophagy:

 

 

https://www.scienced...026049517302494

 

In an earlier iteration of the SC renewal protocol ALCAR has been suggested as promoting neurogenesis i.e. better be taken during the fusion phase?

Perhaps Turnbuckle can weigh in which effect is more pronounced.



#988 Nate-2004

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Posted 22 June 2018 - 05:57 PM

In an earlier iteration of the SC renewal protocol ALCAR has been suggested as promoting neurogenesis i.e. better be taken during the fusion phase?

Perhaps Turnbuckle can weigh in which effect is more pronounced.

 

ALCAR promotes autophagy and fat burning.

 

https://www.scienced...026049517302494

https://nutritionj.b...475-2891-13-110


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#989 Turnbuckle

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Posted 22 June 2018 - 06:32 PM

In an earlier iteration of the SC renewal protocol ALCAR has been suggested as promoting neurogenesis i.e. better be taken during the fusion phase?

Perhaps Turnbuckle can weigh in which effect is more pronounced.

 

 

Carnitine and ALCAR will likely inhibit fission, and thus would go better with fusion. And given the usefulness of carnitine shown in high fat diets, both carnitine and ALCAR would seem excellent additions to fasting. As in the red light fast, since the fuel source during fasting is the triglycerides released by fat cells. 


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#990 Rocket

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Posted 23 June 2018 - 12:39 AM

So what are you guys getting out of this? Better looking skin? Better endurance? More strength?

Can someone quantify the results? I am in the process of sourcing the products but want to hear real stories before buying all this new stuff to start a new stack of sups. Will it aid me in weight training and how?

Edited by Rocket, 23 June 2018 - 12:40 AM.

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