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Manipulating mitochondrial dynamics

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

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#991 HaplogroupW

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Posted 23 June 2018 - 05:38 PM

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. 

 

Good points, and thanks for the ref on inhibiting fission.

 

My main motivation for bringing this up was that I have the sense that there might be a missing ingredient in this protocol. Namely, something to drive mitophagy. We know people who are chronically overfed/have metabolic syndrome/ pre-diabetes/diabetes are biased towards fission and fragmented mitochondria, but are still metabolically diseased. So merely having fission doesn't necessarily mean mitophagy will come along to clean up the detritus of diseased mitochondria. Presumably this can be due the down-regulation of autophagy in a hyper-caloric environment. Maybe the NAM already covers this. Or perhaps there needs to be some element of fasting or something else to drive mitophagy to make it complete. Your point about ALCAR not a fit to fission phase and moving to a fasting period is well-taken.


Edited by HaplogroupW, 23 June 2018 - 05:45 PM.

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#992 Turnbuckle

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Posted 23 June 2018 - 06:49 PM

Good points, and thanks for the ref on inhibiting fission.

 

My main motivation for bringing this up was that I have the sense that there might be a missing ingredient in this protocol. Namely, something to drive mitophagy. We know people who are chronically overfed/have metabolic syndrome/ pre-diabetes/diabetes are biased towards fission and fragmented mitochondria, but are still metabolically diseased. So merely having fission doesn't necessarily mean mitophagy will come along to clean up the detritus of diseased mitochondria. Presumably this can be due the down-regulation of autophagy in a hyper-caloric environment. Maybe the NAM already covers this. Or perhaps there needs to be some element of fasting or something else to drive mitophagy to make it complete. Your point about ALCAR not a fit to fission phase and moving to a fasting period is well-taken.

 

 

There are a couple of other things needed for mitophagy and some may be deficient in them. See this post from the ancestor thread to this one.


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#993 sub7

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Posted 24 June 2018 - 10:58 AM

Why not add Melatonin to your arsenal of mitochondrial manipulators?

(file attached)

 

Quoted from the attached file:

"In most cases,melatonin reduces mitochondrial fission and elevates their fusion. Mitochondrial dynamics exhibitan oscillatory pattern which matches the melatonin circadian secretory rhythm in pinealeocytes andprobably in other cells. Recently, melatonin has been found to promote mitophagy and improve homeostasis of mitochondria"

Attached Files


Edited by sub7, 24 June 2018 - 11:03 AM.

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#994 Turnbuckle

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Posted 24 June 2018 - 12:54 PM

 

"In most cases,melatonin reduces mitochondrial fission and elevates their fusion. Mitochondrial dynamics exhibitan oscillatory pattern which matches the melatonin circadian secretory rhythm in pinealeocytes andprobably in other cells. Recently, melatonin has been found to promote mitophagy and improve homeostasis of mitochondria"

 

Fusion is inconsistent with mitophagy, which requires fission.


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#995 Rocket

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Posted 24 June 2018 - 01:27 PM

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?

 

No one has any experiences or results to share? 



#996 sub7

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Posted 24 June 2018 - 03:02 PM

Fusion is inconsistent with mitophagy, which requires fission.

 

In that case the paper is all wrong. 

Why don't you contact the authors and have that corrected? 


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#997 HaplogroupW

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Posted 24 June 2018 - 07:04 PM

Fusion is inconsistent with mitophagy, which requires fission.

 

My reading so far suggests it would be more accurate to say fission must have happened at some point prior to mitophagy, having left behind fragmented, low- or no-membrane potential mitochondria. These fragmented diseased mitochondria have a reduced or non-existent ability to fuse, and might not get cleaned up until the next time mitophagy is ramps up at some later time.

 

So while fission is required for mitophagy, they don't need to happen at the same time.

 

Your statement, taken naively at face value seems to imply that mitophagy and fusion could never happen simultaneously, which I think would clearly be wrong. I don't think there's any doubt (the references have already been posted here and on other threads) that 1) nutrient starvation promotes mt fusion, and 2) nutrient starvation upregulates autophagy. In fact I recall the authors in one paper proposing that mt fusion is an evolutionary adaptation to prevent the mitochondria all being destroyed during periods of starvation. The autophagosome is physically unable to engulf  tubular or filament-like mitochondria because they are too large.

 

I don't know any reason why melatonin couldn't be having a similar effect to starvation, especially in view of sub7's reference.

 

 


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#998 Turnbuckle

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Posted 24 June 2018 - 07:50 PM

I'll state it more directly: fission is required for mitophagy. 

 

First, mitochondria have to be small enough to fit into lysosomes, therefore they must be fissioned to a size that can be recycled. Second, mtDNA loops cover for each other, so mitochondria have to be fissioned to the smallest size--preferably with one mtDNA loop each--so that low ΔΨm is noticed and these mitochondria suitiably labeled. Every single mtDNA loop in a cell could have a defective gene and it could continue on its merry way, thanks to fusion and the small chance of several mtDNA strands in a mitochondrion having the same defective gene. Until those mitochondria are fissioned, they won't get labeled and destroyed.

 

I don't see why anyone would argue with that. If you take a substance that drives mitochondria to fusion, you shut that down. The paper says, "in most cases,melatonin reduces mitochondrial fission and elevates their fusion." So while melatonin is present at some level, you will shut down mitophagy, which includes the identification of defective mitochondria. Any mitochondria already labeled during a previous fission state will still be destroyed.

 

Is this clear enough?


Edited by Turnbuckle, 24 June 2018 - 08:02 PM.

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#999 HaplogroupW

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Posted 24 June 2018 - 11:29 PM

No one has any experiences or results to share? 

 

Rocket- I haven't been measuring anything in a systematic way, and so can't quantify any results. Even if I had, it would be difficult to disentangle the effects of the other interventions I engage in. Subjectively I haven't noticed anything either. It could be because I'm fairly metabolically healthy to begin with. I also don't subjectively experience the decline of aging as it progresses from day to day, and yet it is still real. I have to look at a photo of myself from 20 years ago to notice that I'm getting older. To the extent that I do this protocol or any of the other interventions in which I engage, it's based on a the idea that the mechanisms described in the literature are real,  and perhaps a dose of hope.


Edited by HaplogroupW, 24 June 2018 - 11:36 PM.

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#1000 Rocket

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Posted 25 June 2018 - 01:18 AM

Do you feel anything on fission/fusion days? Turnbuckle said something about low BP and feeling tired.

It would be nice to hear about something that actually worked, on this site.

I had some good experiences with lipsomal resveratrol and bpc157 but I would not call it antiaging.

#1001 pone11

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Posted 26 June 2018 - 11:23 PM

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

 

What sensations or effects do you get on sulforaphane?  How were you taking it and what dose?



#1002 Turnbuckle

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Posted 27 June 2018 - 09:57 AM

What sensations or effects do you get on sulforaphane?  How were you taking it and what dose?

 

 

I would sometimes get nausea and/or flu-like symptoms from taking more than one cap of broccoli sprout extract, even though they had very little sulforaphane -- 1.5 mg/cap. Yesterday I tried broccoli seed extract from Thorne with 50 mg sulforaphane glucosinolate. This amounts to 20 mg sulforaphane per cap, and so far I've experienced only ringing in the ears. If I get the ringing again, I will go back to stearic acid.


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#1003 displayname

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Posted 28 June 2018 - 10:31 PM

What are some sources of Sodium pyruvate/methods of increasing its endogenous production?

(Edit: Ok, just testing if I can still post. For some reason I was restricted before.)


Edited by displayname, 28 June 2018 - 10:35 PM.

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#1004 ementic

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Posted 12 July 2018 - 02:25 AM

do you think this protocol will work well if it is followed to its entirety with the exception of using apigenin? 



#1005 onz

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Posted 12 July 2018 - 04:33 AM

The current protocol calls for 3 days of fission, followed by 2 days of fusion... Is it possible to shorten this to 1 day of fission? I'm getting ready to try the protocol, but my concern is the decrease in energy experienced during fission will be too inconvenient if it's lasting 3 days



#1006 Turnbuckle

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Posted 14 July 2018 - 01:00 AM

It should work without the apigenin, and I would expect that as long as you allowed the stearic acid to leave your system, that one day of fission would achieve something. Then you can go from there depending on the results. If you have a lot of bad mitochondria, it could take many cycles to get rid of them.


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#1007 ementic

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Posted 14 July 2018 - 08:48 PM

I really appreciate your contribution turnbuckle. I've just finished one cycle and the difference has been striking. I'm not used to seeing such clear results with health interventions. Is there any reason I can't continue cycles back to back until I see no improvement?


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#1008 Turnbuckle

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Posted 14 July 2018 - 09:08 PM

I really appreciate your contribution turnbuckle. I've just finished one cycle and the difference has been striking. I'm not used to seeing such clear results with health interventions. Is there any reason I can't continue cycles back to back until I see no improvement?

 

 

You have to. It's an iterative process of destruction and creation, and if you have a lot of bad mtDNA loops, there's no way you could get rid of them after one cycle, or two, or ten. It took me months to get to the point that I felt most of them were gone. At that point the original distinct difference between fission and fusion should seem very minor.


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#1009 ementic

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Posted 17 July 2018 - 02:46 AM

Ok perfect. Last question; I saw you made sure to dissolve the stearic acid on your c60 thread. Is that necessary here or is the dry powder fine?



#1010 Nate-2004

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Posted 20 July 2018 - 07:47 PM

So I've been fasting a lot lately, 48 hours per week and I've been trying to work my way up to 72 hours and ultimately 120 hours per month for maybe 6 months.

 

In terms of dynamics I've heard you suggest that fasting may be more helpful in inducing fusion, not fission. This is despite the suggestion to fast prior to a workout which as I understand it, is for helping deplete ATP during your fission workout protocol not for inducing fission itself. 

 

So, I was going to do another 5 days of fission soon, should I not do that during a fast? Just to note, typically during the fast I'm taking metformin as an AMPK activator which should help with the autophagy. However, don't mitophagy and autophagy go hand in hand or not?

 

Maybe just do fission during a refeed?


Edited by Nate-2004, 20 July 2018 - 07:47 PM.


#1011 Turnbuckle

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Posted 20 July 2018 - 08:18 PM

So I've been fasting a lot lately, 48 hours per week and I've been trying to work my way up to 72 hours and ultimately 120 hours per month for maybe 6 months.

 

In terms of dynamics I've heard you suggest that fasting may be more helpful in inducing fusion, not fission. This is despite the suggestion to fast prior to a workout which as I understand it, is for helping deplete ATP during your fission workout protocol not for inducing fission itself. 

 

So, I was going to do another 5 days of fission soon, should I not do that during a fast? Just to note, typically during the fast I'm taking metformin as an AMPK activator which should help with the autophagy. However, don't mitophagy and autophagy go hand in hand or not?

 

Maybe just do fission during a refeed?

 

 

I've suggested that fasting goes well with fusion, which is in fact a natural response to fasting. With fasting, the problem is dealing with hunger, and hunger is triggered by a shortfall in the energy supply from triglycerides. Fused mitochondria are more efficient at burning triglycerides than fragmented mitochondria, so the triglyceride demand is closer to what your fat cells can supply. You can up the supply with L-carnitine fumarate and red light, preventing hunger altogether.

 

See: losing weight at light speed.


Edited by Turnbuckle, 20 July 2018 - 08:37 PM.

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#1012 Fafner55

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Posted 21 July 2018 - 01:59 PM

More support for the need to keep populations of mitochondria healthy.

 

"Reversing wrinkled skin and hair loss in mice by restoring mitochondrial function" (2018) https://www.nature.c...1419-018-0765-9

Mitochondrial DNA (mtDNA) depletion is involved in mtDNA depletion syndromes, mitochondrial diseases, aging and aging-associated chronic diseases, and other human pathologies. To evaluate the consequences of depletion of mtDNA in the whole animal, we created an inducible mtDNA-depleter mouse expressing, in the polymerase domain of POLG1, a dominant-negative mutation to induce depletion of mtDNA in various tissues. These mice showed reduced mtDNA content, reduced mitochondrial gene expression, and instability of supercomplexes involved in oxidative phosphorylation (OXPHOS) resulting in reduced OXPHOS enzymatic activities. We demonstrate that ubiquitous depletion of mtDNA in mice leads to predominant and profound effects on the skin resulting in wrinkles and visual hair loss with an increased number of dysfunctional hair follicles and inflammatory responses. Development of skin wrinkle was associated with the significant epidermal hyperplasia, hyperkeratosis, increased expression of matrix metalloproteinases, and decreased expression of matrix metalloproteinase inhibitor TIMP1. We also discovered markedly increased skin inflammation that appears to be a contributing factor in skin pathology. Histopathologic analyses revealed dysfunctional hair follicles. mtDNA-depleter mice also show changes in expression of aging-associated markers including IGF1R, KLOTHO, VEGF, and MRPS5. mtDNA-repleter mice showed that, by turning off the mutant POLG1 transgene expression, mitochondrial function, as well as the skin and hair pathology, is reversed to wild-type level. To our knowledge that restoration of mitochondrial functions can reverse the skin and hair pathology is unprecedented.


Edited by Fafner55, 21 July 2018 - 02:16 PM.

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#1013 Rocket

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Posted 22 July 2018 - 12:36 AM

Fasting? Fasting??? Are you trying to lose muscle with all that food avoidance?
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#1014 Nate-2004

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Posted 22 July 2018 - 01:44 PM

Fasting? Fasting??? Are you trying to lose muscle with all that food avoidance?

 

Not sure if this is sarcastic or what lol

 

Fasting... yeah.... you know, the thing being talked about most these days in longevity circles. And no that's not what causes muscle loss unless you're out of fat.


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#1015 Rocket

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Posted 23 July 2018 - 08:07 PM

Nate...….…………

 

I've read your posts and I know that you train with weights. I thought you surely recognized the value in building muscle as you get older to improve your quality of life in what are for most people the "frail" years where they can't open a soup can or walk upstairs. The more muscle now while you're young the better.

 

As long as you're essentially dieting, you're never going to make any gains in lean muscle or gains in the gym. Fasting as much as you are talking about doing is 100% totally counterproductive to weight training and making gains. You may even start... or eventually will slip backwards as sarcopenia robs you of muscle each and every year as it does to all humans.

 

Also if CRON doesn't translate into humans like mice, then intermittent fasting can be considered CRON-LITE and won't add decades to your life...… It just doesn't work in humans. Consensus seems to be something like 2 additional years. 

 

EDIT: I know fasting improves some biomarkers, but big whoop if you're skinny and weak and old. My $0.02.

 

 

 

 

 

 


Edited by Rocket, 23 July 2018 - 08:08 PM.

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#1016 Turnbuckle

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Posted 23 July 2018 - 09:20 PM

The more muscle now while you're young the better.

 

 

 

Are you sure? I've seen a lot of jocks from my high school days turn into rotten pumpkins over the years, so anecdotally it seems that muscle building when young might make it more difficult to build muscle when elderly. Which makes sense when you consider that muscle cell function is dependent on muscle stem cells. These are called satellite cells as they reside outside the muscle fibers, and once activated migrate into fibers and replenish their nuclei. Like other stem cells, constant asymmetric division (differentiation) will result in a decline in numbers and function. In fact, the majority of muscle wasting in the elderly is caused by the loss of satellite cell function. 

 

Satellite cells are a heterogeneous collection of adult muscle stem cells that are normally quiescent. ... Adult satellite cells remain quiescent and reside (relatively) dormant within their niche adjacent to the myofiber. While satellite cells might be exposed to the changing cellular niche, they do not become activated until a major insult or stress (e.g., exercise loading) occurs. In response to injury, satellite cells proliferate and their Pax7-positive daughter cells either differentiate, by migrating through the sarcolemma and fusing with existing muscle fibers ... during the growth and regeneration of muscle... or they commit to a program of self-renewal.

 

 

 
So the wasting of muscles with age is due to the decline of the functional satellite cell pool. To reverse that, you'd like to enlarge that pool (actually >600 pools for the >600 muscles) by biasing satellite cells to self-renewal (symmetric division rather than asymmetric, where one satellite cell becomes two). Proliferation of satellite cells is begun by a unique trigger -- nitric oxide -- by which these cells detect muscle damage that needs to be repaired. You can fake them out with supplements like potassium nitrate, which produces that nitric oxide signal. And if satellite cells act like other stem cells in other respects, they can be biased to self renewal by mitochondrial fusion with stearic acid. I created a protocol using stearic acid and potassium nitrate to do this, which can be found at the bottom of this post -- Satellite cell self-renewal, without C60.

Edited by Turnbuckle, 23 July 2018 - 09:29 PM.

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#1017 sthira

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Posted 24 July 2018 - 04:14 AM

Friendly conversation ahead...

...As long as you're essentially dieting, you're never going to make any gains in lean muscle or gains in the gym. Fasting as much as you are talking about doing is 100% totally counterproductive to weight training and making gains. You may even start... or eventually will slip backwards as sarcopenia robs you of muscle each and every year as it does to all humans.


Aren't muscle gain and muscle loss the results of exercise or lack of exercise, and not diet or fasting?

And why would we burn muscle and bone when fasting, rather than stored fat?

Consider: https://www.ncbi.nlm...ubmed/20300080/

Here, researchers studied humans who underwent 70 days of alternate daily fasting (ADF: eat one day; fast the next). And their fat free mass started at 52.0 kg, and ended at 51.9 kg. They didn't lose lean weight (bone, muscle.), they lost fat.

Fat exists, in part, to be utilized during times of food scarcity. Why would the human body, evolved as it has over hundreds of thousands of years, store excess energy as fat, and then right turn around and burn up protein when starving?

Protein is functional tissue -- it's bone, it's muscle, and it's needed to help us find food.

Fat is stored energy for times when we can't; stored excess fat is how our ancestors survived starvation. Nate will burn fat when he fasts, he won't burn much muscle and bone.

And we aren't the only animals that burn fat for energy when fasting. All animals do it -- we eat our own internally stored fat. Animals don't waste muscle and bone, and especially we don't waste muscle and bone while simultaneously holding onto fat stores when starving and forced to use muscles and bones when foraging and hunting.

Also if CRON doesn't translate into humans like mice, then intermittent fasting can be considered CRON-LITE and won't add decades to your life...… It just doesn't work in humans. Consensus seems to be something like 2 additional years.

EDIT: I know fasting improves some biomarkers, but big whoop if you're skinny and weak and old. My $0.02.


Researchers can't study the effects of CR on human lifespan extension because it's too hard. But going out on a limb here, CR extends lifespan in just about every other species tested. So why would humans be exceptional? Why would we, of all other species, fail to have adapted mechanisms to extend lifespan via CR, fasting, and food deprivation?

We did, you say, but we only get two extra years. Where are you getting the idea that consensus says CR extends human lifespan by two years? What consensus?
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#1018 Nate-2004

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Posted 24 July 2018 - 05:55 PM

Aiming for 120 hours this week, I'm nearly at 48 now, dunno if I'll make it past 48 or not lol. I am hoping to just do 120 a month, 48 a week is not fun. The N-Acetyl-L-Carnitine is helping a lot in terms of hunger and making energy. I also get into ketosis really fast, by 48 hours I'm at 1.2 on the blood meter. From what I've read by people who don't cite sources, that's when autophagy/mitophagy begins. Mitophagy is what we're aiming for in this thread buuuuut...

 

Here's what I still don't understand, if fasting leads to autophagy and also mitophagy, yet fasting also induces fusion, then how is mitophagy happening? If they're fused, then they're not going to be eaten up by lysosomes. So then if I'm doing a fission protocol then how am I going to induce mitophagy? Is it the ATP depletion via resistance exercise? Earlier in this thread it was recommended to fast up until an hour after a workout during fission. Why exactly? How does that not work against the fusion response to fasting?

 

Also, with dynamics manipulation in consideration, how should I end the fast? With stearic acid, glutathione and C60 (per the stem cell thread)? Or should I just end it normally like I do with a big dose of electrolytes, thiamin, and a fiber packed smoothie with walnuts, flaxseed, sulforaphane induced raw broccoli (i.e. warm water at 60c and blended, stirring frequently till cool), inulin, whole milk yogurt and blueberries?  

 

Would going from extended fasting into a fission protocol be bad or good?

 

I really wish they'd study fasting more.  I don't know why but the 48 hours a week is definitely helping with my tremors.

 


Edited by Nate-2004, 24 July 2018 - 05:58 PM.


#1019 Turnbuckle

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Posted 24 July 2018 - 06:05 PM

Here's what I still don't understand, if fasting leads to autophagy and also mitophagy, yet fasting also induces fusion, then how is mitophagy happening? If they're fused, then they're not going to be eaten up by lysosomes. So then if I'm doing a fission protocol then how am I going to induce mitophagy? Is it the ATP depletion via resistance exercise? Earlier in this thread it was recommended to fast up until an hour after a workout during fission. Why exactly? How does that not work against the fusion response to fasting?

 

 

 

It's like this--

 

A fundamental role for mitochondrial dynamics during mitophagy is also supported by the fact that acute fasting promotes hyper-fusion of mitochondria via protein kinase A mediated inhibition of Drp1. This hyperfused state of mitochondria is thought to prevent mitochondrial degradation by mitophagy during short periods of nutrient limitations although mitochondria are eliminated following chronic starvation conditions...

https://www.ncbi.nlm...les/PMC3969632/

 

 


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#1020 Nate-2004

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Posted 24 July 2018 - 07:12 PM

It's like this--

 

A fundamental role for mitochondrial dynamics during mitophagy is also supported by the fact that acute fasting promotes hyper-fusion of mitochondria via protein kinase A mediated inhibition of Drp1. This hyperfused state of mitochondria is thought to prevent mitochondrial degradation by mitophagy during short periods of nutrient limitations although mitochondria are eliminated following chronic starvation conditions...

https://www.ncbi.nlm...les/PMC3969632/

 

I actually went and read the full context. Still doesn't quite confirm either way. So mitophagy doesn't happen during fasts? They seem to imply that the fusion is brief (during short periods), but they don't mention how brief.







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