2287 replies to this topic

[Adam1]

Many arrhythmias cause no signs or symptoms.

[PAMPAGUY]

Have now done 4 cycles of fission - fusion started out with 1.5N + 3R x 3 days. Big mistake. Kicked my butt. Next week 1N 2R x3. Better but last day was hard. Tried 1N 2R x 2 days. Felt good. Will go slow like Turnbuckle recommended. Building up slowly to 1.5N + 3R x3. Fusion was no problem. Day off between fission and fusion. Used 10g stearic acid+20mg PQQ+Broccomax 1 capsule on 3 fusion days. Used stearic acid 10g 1st. 2 days then 5g last day.  Lots of energy except when I overdue it.

Would appreciate any help with my protocol.

Edited by PAMPAGUY, 26 December 2017 - 08:55 AM.

[Andey]

Have now done 4 cycles of fission - fusion started out with 1.5N + 3R x 3 days. Big mistake. Kicked my butt. Next week 1N 2R x3. Better but last day was hard. Tried 1N 2R x 2 days. Felt good. Will go slow like Turnbuckle recommended. Building up slowly to 1.5N + 3R x3. Fusion was no problem. Day off between fission and fusion. Used 10g stearic acid+20mg PQQ+Broccomax 1 capsule on 3 fusion days. Used stearic acid 10g 1st. 2 days then 5g last day.

Would appreciate any help with my protocol.

If I recall correctly it's a way higher dosage than one used to kick your but before. If the previous dosage doesn't cause it now than you are on a right track and you mito population is healthier than before.

I monitor my overreaching after night fission protocol with morning BG levels that reflects cortisol levels = stress, and it shows that body adapts to fission protocol slowly but steadily to the level when I consider adding more to the mix.

[PAMPAGUY]

Andey thanks for sharing and your right I can tolerate higher dose. My big problem before was taking 1000 mg Metformin with Fission. Turnbuckle advised it was an uncoupler. Stopped on fission days and have been handling much better.

[BieraK]

Report here, an experimental modified fission protocol for removing mutated mitochondria:
Before bed I took
4 grams N + 5 grams R
1000 mg Cyelon Cinnamon

------------------------------------------

This for increasing mitophagy and fission

After 30 minutes
1 gram Aspirin

---------------------------------------------

A bit dangerous, concerns of bleeding. I took aspirin for being an uncoupler in such doses

Aproximately 1 hour after the N+R dose I took 100 mg Gypenosides for AMPK and increased fission.

Then I do HIIT sessions on the elliptical bike. I was able only to do 6 x 25 seg hight intensity - 10 seg low intensity.
Then I go to bed, I felt tired before doing HIIT, and after it was enhanced, I was trying to read in the notebook however I was feeling tired and a bit body uncomfort, so I decided to sleep. Sleeping was too profound, like a melatoning dose, I dreamed a lot also. 

I've never used such doses of aspirin, so I don't know how is the anti-inflammatory potential, but I felt with less nerve and head pain in my neck and a general sense of well being, tired but with a sense of wellbeing. 

Today I'm feeling refreshed, a bit "foggy" and sleepy. I'm feeling my heart calmer, I think that this stack dropped my blood pressure now is in 100 - 60. While sleeping I was feeling an strange sensation like being too slow and calmer I think that it was a drop in blood pressure by that moment, however I was not feeling groggy the two times I wake up in the night.

So interesting stack, this is not the first time I do it. The difference now was the inclussion of a mild uncoupler. It is difficult for me to know if the differences compared to regular fission protocol are due to reduced mutated mitochondrias or just for the effects of aspirin per se, I don't wan't to use more aspirin for uncoupling because looks too risky, I don't have access to DNP so perhaps metformin could be another optio. Now I will go for the Sulforaphane, PQQ and a high dose of pure Cacao Powder.

Edited by BieraK, 27 December 2017 - 04:42 PM.

[PAMPAGUY]

I was taking Metformin unknowingly during Fission and it kicked my ass. 1000mg
It is a powerful uncoupler.

[Andey]

I imagine that ketogenic diet + metformin + periodic N+R could be a good anticancer therapy (or a prevention measure). At least against glycolytic cancer types

[Adam1]

I don't know if the month-long silence on the thread was related to issues I raised at the end of November that were not discussed. Now that the thread is showing signs of life, I will reflect on this. It’s one thing to promote a protocol for mitochondrial dynamics if it is harmless, but I pointed out the protocol can kill if it works, and the response was to walk away rather than address the evidence.

 

Not only, as I posted above, can inadvertently induced mitochondrial dysregulation cause organs to collapse, but also as studies show it can bring about massive loss of neurons (neurodegenerative diseases such as Parkinson’s) etc. This potential damage starts invisibly.

 

Mitochondrial dynamics are rapid and multifaceted, and include fleeting kiss-and-run interactions that require flexibility and responsiveness. Thus, any protocol that constrains dynamics and entrains slow cycles of fission and fusion should consider safety and likelihood of improving dynamics. Is there even one shred of evidence that dynamics are helped by slowing the vibrancy of natural oscillations to a relative dead halt?

[Richard McGee]

I don't know if the month-long silence on the thread was related to issues I raised at the end of November that were not discussed. Now that the thread is showing signs of life, I will reflect on this. It’s one thing to promote a protocol for mitochondrial dynamics if it is harmless, but I pointed out the protocol can kill if it works, and the response was to walk away rather than address the evidence.

 

Not only, as I posted above, can inadvertently induced mitochondrial dysregulation cause organs to collapse, but also as studies show it can bring about massive loss of neurons (neurodegenerative diseases such as Parkinson’s) etc. This potential damage starts invisibly.

 

Mitochondrial dynamics are rapid and multifaceted, and include fleeting kiss-and-run interactions that require flexibility and responsiveness. Thus, any protocol that constrains dynamics and entrains slow cycles of fission and fusion should consider safety and likelihood of improving dynamics. Is there even one shred of evidence that dynamics are helped by slowing the vibrancy of natural oscillations to a relative dead halt?

 

It is indeed wise to exercise caution when dealing with this, or any other, untested protocol. That being said, all life is risk, including the risk of missed opportunities that might improve one's health. The risk of missed opportunities is especially poignant, given 1) the the halting slowness of infiltration of medical research into application, 2) the dearth of applications that can't be easily converted into profit, and 3) the limited time that we have as individuals to deal with these issues.

 

These factors change the risk/reward analysis for many people. This doesn't mean throwing caution to the winds. It means playing close attention to our body's reactions. It means becoming as well-informed as possible on the scientific rationale behind these protocols. It means paying attention to new information as it become available, including any negative indications.

 

Mitochondrial dysregulation is a fact of life as we age. The "potential" damage is actually very real and accumulates throughout life, and you're certainly not going to avoid it by doing nothing. In the absence of controlled studies we can't actually specify with any degree of certainty how this protocol works at the cellular level. However, I'm skeptical of your assertion that we are "slowing the vibrancy of natural oscillations to a relative dead halt." It is equally likely that we are simply weighting the likelihood of fission or fusion in the aggregate, not halting the fleeting micro-interactions. This seems more in line with the reported effects described by those who have utilized the protocols. 

 

Nevertheless, caution is justified and we should be on the lookout for any reported negative effects.

[Adam1]

Thanks Richard. I was not suggesting to do nothing. There are alternatives.

 

Someone indicated on my last post they wanted references. Please specify what you need backed up.

 

 

[StanG]

I'm delighted to see you post again Adam1. This blog started out well but it went overboard IMO quite awhile ago and I stopped following it. It seemed as though everybody was trying to out do the last one. We all take chances with what we do and I'd hate to see anyone harmed by what they do but when I read about effects some of these people have, I have to wonder what will happen to them in the long run if they continue what they're doing.

 

I don't expect this to be popular but if it stops one person from overdoing things, it served its' purpose. 

Edited by StanG, 27 December 2017 - 10:22 PM.

[Adam1]

Thanks Stan. You understand where I am coming from.

[Nate-2004]

On the subject of uncoupling during fission, earlier in the thread I mentioned that berberine inhibits uncoupling with a link to a study concluding as much, but now I read here in another publication that berberine induces thermogenesis, which I assume means uncoupling. Berberine obviously needs more study, what with these kinds of contradictions.

[Adam1]

Berberine is one of the many compounds that induce both autophagy and biogenesis. Referenced here.

[Andey]

I don't know if the month-long silence on the thread was related to issues I raised at the end of November that were not discussed. Now that the thread is showing signs of life, I will reflect on this. It’s one thing to promote a protocol for mitochondrial dynamics if it is harmless, but I pointed out the protocol can kill if it works, and the response was to walk away rather than address the evidence.

 

Not only, as I posted above, can inadvertently induced mitochondrial dysregulation cause organs to collapse, but also as studies show it can bring about massive loss of neurons (neurodegenerative diseases such as Parkinson’s) etc. This potential damage starts invisibly.

 

Mitochondrial dynamics are rapid and multifaceted, and include fleeting kiss-and-run interactions that require flexibility and responsiveness. Thus, any protocol that constrains dynamics and entrains slow cycles of fission and fusion should consider safety and likelihood of improving dynamics. Is there even one shred of evidence that dynamics are helped by slowing the vibrancy of natural oscillations to a relative dead halt?

 

    Do you have any evidence to start with that protocol "slowing the vibrancy of natural oscillations to a relative dead halt"?

  Looks like you are opposing not the protocol but your idea of what protocol is instead. Compare it to fasting or exercise or whatever that shifts mitochondrial dynamics equilibrium. Probably its effects more profound than any natural approach but I hardly doubt its so profound as you speak. If you look at collective experience the more advanced age somebody is, the more profound side effects are experienced generally. I for one hardly feel anything and resort to measuring HRV, BG and ketones to monitor its effects. If you are right then I should feel at least something when all mitochondria in all cells in my body strangled, is in it?

Seriously, I am not saying that its safe 100%, far from it. If protocol kicks relatively healthy individuals, it could be too little too much for not so healthy ones. As too much exercise or fasting could lead to heart attack and kill if somebody is already 99% down the road to this outcome. 

Edited by Andey, 28 December 2017 - 07:16 AM.

[Turnbuckle]

I don't know if the month-long silence on the thread was related to issues I raised at the end of November that were not discussed. Now that the thread is showing signs of life, I will reflect on this. It’s one thing to promote a protocol for mitochondrial dynamics if it is harmless, but I pointed out the protocol can kill if it works, and the response was to walk away rather than address the evidence.

 

Not only, as I posted above, can inadvertently induced mitochondrial dysregulation cause organs to collapse, but also as studies show it can bring about massive loss of neurons (neurodegenerative diseases such as Parkinson’s) etc. This potential damage starts invisibly.

 

Mitochondrial dynamics are rapid and multifaceted, and include fleeting kiss-and-run interactions that require flexibility and responsiveness. Thus, any protocol that constrains dynamics and entrains slow cycles of fission and fusion should consider safety and likelihood of improving dynamics. Is there even one shred of evidence that dynamics are helped by slowing the vibrancy of natural oscillations to a relative dead halt?

 

You are welcome to your opinion, but this is a site for experimenters, and people know that. Your concerns have been noted, but repeated over the top warnings we can do without. As for this "vibrancy of natural oscillations," that is too kooky to address.

[Nate-2004]

The silence was more due to the holidays than the arguments you were making. Honestly, I didn't find them convincing enough to be alarmed by them. Turnbuckle is right, this is a site for experimenters. There's no supporting research or evidence for any claims made in this thread about manipulating mitochondrial dynamics. It's all hypothesis based on a review of existing evidence about how the quality control system works. The hypothesis is all we can really experiment with until a legitimate controlled study involving more participants is run.

 

I don't think anybody trying this protocol is going to drop dead any time soon. We're all pretty healthy as is just based on our lifestyle. I have to assume that since there is a strong interest in longevity here. Cycling supplements is nothing new and not mixing one supplement with another, is also nothing new. Trying to avoid contradicting interactions is nothing new. These things are not going to kill us. In fact I've never felt better.

[Adam1]

Thanks for the responses. This cycling protocol was designed with one purpose: “to increase mito fission or fusion for specific goals.” The protocol has alternating fission days and fusion days, with an emphasis on keeping the two functions separate.  

 

Two possibilities:

1. The protocol does what it was designed to do  

Most of my expressed concern: if the protocol guides fission and fusion as it was meant to, there may be some potentially serious issues. The cycles of functional mitochondrial dynamics are extremely rapid. Some of these functional cycles need to include partial and transient fusions where only a few proteins are exchanged (post above referencing kiss-and-run).

 

In contrast, the protocol is for dedicated fission days and fusion days. As noted, Turnbuckle said “expect that fluidity is not lost, but the center-point is pushed first in one direction, then the other--a slow oscillation that operates in the background to the rapid activity of mitochondria.” I did not get an answer on: “what were you saying was oscillating slowly?” If there is nothing that can even be speculated about in the background, I have to conclude that the mitochondria are influenced directly if the protocol works. You can’t say the protocol exclusively promotes fission one day and fusion another day and yet can’t have an effect on the speed and flexibility of dynamics.

 

To be fair, there may be links to circadian rhythm. For example, a study published last month suggests it is possible mitochondria are driven by a roughly 24 hour rhythm concurrently with rapid dynamics. Some of the preliminary evidence for such is that mitochondria appear to be entrained by light:

 

The primary site of light absorption in mammalian cells has been identified as the mitochondria, and more specifically, cytochrome c oxidase (CCO). It is hypothesized that inhibitory nitric oxide can be dissociated from CCO thus restoring electron transport and increasing mitochondrial membrane potential. Another mechanism involves activation of light or heat-gated ion channels.

 

However, this protocol does not revolve around this circadian hypothesis, and appears to be unaware of it, so post-hoc rationalizations would be questionable. Besides, the above circadian research is consistent with the understanding that mitochondria are highly responsive to their environment and entrained by it.

 

Even if as Turnbuckle explained, the protocol was not for the mitochondria but for his proposed unspecified background process, the mitochondria could be entrained by this process. In my posts last month I referenced evidence that mitochondria are organized as a network susceptible to global entrainment. The risk of entrainment reaching criticality and system collapse is analogous to sudden seizures after a period of strobe light exposure in photosensitive epilepsy.

 

One of the key reasons for cycling stacks is to prevent homeostatic adaptations to any regimen, thus to keep the intervention effective. This protocol is different from common cycling regimens because it narrowly targets the biological cycle of fission and fusion on alternate days, so it could entrain fission and fusion to follow the same slow pattern that is repeatedly followed.

 

    2.   The protocol does not do what it was designed to do

 

Events following critical thresholds don’t always have warning signs. However, it is possible those who are not feeling ill effects are actually safe because the protocol does not work in the ways that it is meant to work.

 

Exercise promotes biogenesis, fusion, fission, and mitophagy (attached picture):

 

 

 

There was a question on this thread: “This protocol calls for inducing mitochcondrial biogenesis on fusion days. Is NR itself, taken on fission days, the heavyweight though when it comes to biogenesis?” This question was referenced with one study that Turnbuckle acknowledged: “Ultimately this would be a problem as mitophagy requires fission.” However, there are also other studies and reviews implicating NR in biogenesis, e.g. here, here and here. In addition NR may have a synergistic role in biogenesis (attached picture):

 

 

 

Stearic acid has diverse effects aside from fusion, e.g. this study: “not only the deleterious impact of stearic-acid-induced lipotoxicity but also apoptosis in beta cells and progression to type 2 diabetes” There are age differences in baseline levels of stearic acid according to this study, and this may be a factor in what you observed about age effects, Andey (“our data demonstrated a significant correlation between concentration of palmitic, stearic, and linoleic acid and plasma glucose levels and glycated hemoglobin with aging”).

 

Please let me know if I neglected to respond to anything, or if I am missing something. I don’t know if the protocol works as intended under specific conditions, but it stimulates mitochondrial gateways that can go either way. It looks like the conceptual model behind this protocol may be flawed. I understand self-experimenters go on theory a lot of the time without evidence for the practical application of the theory, but I don’t think the theory is sufficient to support any protocol.

[BieraK]

That's something I've asked myself since I started reading this post. Mitophagy need to be paired with biogenesis?. As I understand CR increases autophagy and mitogenesis, and at a some extent mitophagy. I also wonder what are the effects of NR in low doses compared to high doses, apparently at high doses it induces fussion, but what happens at low doses? It is something that I pointed out here and was answered here
 

Similar statements can be found in other studies

https://www.hindawi....l/2017/2012798/
"It is important to highlight that many interventions leading to health improvement and extension in lifespan, such as calorie restriction, or treatment with rapamycin, spermidine, metformin, or the antioxidant resveratrol, also induce an activation of the autophagic/mitophagic machinery [4]. In addition, regular physical activity has demonstrated benefits on adults’ health and has been also identified as an inducer of autophagy in vivo [2, 10]. In the same line, recent evidences have demonstrated that exercise is also able to affect the activity and turnover of mitochondria by increasing biogenesis and mitophagy [11]"

 

So, one might ask, if not it simpler to remove damaged mitochondria by doing periodic ketogenic diets or periodic CR?
However, it could be thought that CR or mitophagy with biogenesis only "masks" the problem, and not get rid of mutated mitochondria, similar to ketogenic diet that produces biogenesis and enhances autophagy. I not deny at all some repairing effects of CR, Fasting and Keto on mitochondrias and other things, however I don't think that eliminates at all mutated mitochondrias acquired through life and maternal inheritance

 

That's why I'm interested in uncouplers during enhanced fission and mitophagy, the idea is to try to eliminate mutated mitochondria even those inherited. For me DNP looks interesting, obviously in a low dose like 50 mg.
 

I was taking Metformin unknowingly during Fission and it kicked my ass. 1000mg
It is a powerful uncoupler.

I will try (N+R)+Cinnamon+Metformin+Gypenosides the next time, I don't want to take excess Aspirin. For me Cinnamon looks interesting also, because it could enhance mitophagy.

 

 

I imagine that ketogenic diet + metformin + periodic N+R could be a good anticancer therapy (or a prevention measure). At least against glycolytic cancer types

I was doing ketogenic diet for 3 weeks this year, It was wonderful, plenty of energy, focus, motivation, good mood, better skin, less hair loss.
 

I had my doubts about whether to consume N + R with Keto, I assumed that ribose could get me out of ketosis, but perhaps this protocol works better in a state of ketosis due to the enhanced autophagy and mitophagy of ketoD

 

Edited by BieraK, 30 December 2017 - 01:39 AM.

[Turnbuckle]

 

Most of my expressed concern: if the protocol guides fission and fusion as it was meant to, there may be some potentially serious issues. The cycles of functional mitochondrial dynamics are extremely rapid. Some of these functional cycles need to include partial and transient fusions where only a few proteins are exchanged (post above referencing kiss-and-run).

 

 

 

Your kiss-and-run paper says this--

 

Cells expend considerable effort to continually merge and divide mitochondrial compartments that are essential for maintaining the metabolic function of these organelles as well as regulating their roles in cell signalling (Yaffe, 1999; Westermann, 2002; Chan, 2006; McBride et al, 2006; Tatsuta and Langer, 2008). Although it is unclear whether the causal link is to morphology, per se, or to the morphology-regulating proteins, most studies linked loss of fusion with mitochondrial dysfunction and greater susceptibility to apoptosis (Youle and Karbowski, 2005).

 

 

The referenced Youle opinion paper says this--

 

We propose that mitochondrial fission is an

integral step of apoptosis. First, mitochondrial

fragmentation occurs during numerous pathways

of apoptosis in mammalian cells and

during developmental cell death in C. elegans

upstream of caspase activation. Second, two

physiological mediators of mitochondrial fission,

DRP1 and FIS1, mediate this apoptotic

process and are required for the full induction

of cell death. The colocalization of DRP1

with BAX and BAK in submitochondrial

foci further implicates mitochondrial fission

in apoptosis. Early in apoptosis, both DRP1

and BAX move from the cytosol to the same

mitochondrial fission sites.

However, mitochondria divide and fuse

regularly in healthy cells. Reversible mitochondrial

fragmentation can be induced

without inducing apoptosis by, for example,

inhibiting respiratory complexes and raising

cytosolic Ca2+ levels. So, fission per se seems

not to induce apoptosis. We suggest that the

translocation of BAX and BAK to the fission

sites along with DRP1 might be essential to

enable the fission machinery to promote

apoptosis.

 

 

 

Fission is part of the apoptosis process, but is not sufficient to cause it. Also, fusion does not completely stop apoptosis if the cell is marked for it. From the same paper--

 

Although

mitochondrial fragmentation obviously

occurs during apoptosis, the effects of inhibiting

apoptosis by inhibiting mitochondrial

fission — which would demonstrate the crucial

nature of this step for caspase activation

— are mild in some reports. 

 

 

 

Caspases are proteins that execute programmed cell death when activated in a cascade. Without them, the cell is not going to self-destruct. And with them, fusion is not necessarily going to prevent self-destruction. Thus your concern is misplaced. Things that start the caspase cascade include "IR [ionizing radiation] chemotherapeutic drugs, death receptors-mediated processes like tumor necrosis factor α [TNFα], growth factor withdrawal, loss of cell adhesion (anoikis) and cytoskeletal damage..." Source.

Edited by Turnbuckle, 30 December 2017 - 02:46 AM.

[PAMPAGUY]

Don't know if this clinical trial of NR has been posted, but here it is.

 

http://journals.plos...&type=printable

[Nate-2004]

There's just not enough information to draw any conclusions about this. Yes this protocol of cycling NAM+R or NR is targeted less towards adaption and more towards fission/fusion but as far as #1 or #2 as far as Adam1 puts it, I can't begin to have any certainty about it. I'm more inclined to revert back to my previous protocol of morning NR and fast and then 2 hrs after post workout sulforaphane and antioxidants with food. Separated by hours not days. 

 

I don't know for sure that there is any real way to get QC on dysfunctional or mutated mitochondria but it's one of the major targets in aging research, one in which SENS proposes a solution of storing "backup copies" (see point 3) in the cell nucleus, rather than attempting to weed out the baddies so to speak. I think this is something I'll have to read more about but perhaps there's just not enough information to draw any actionable conclusions right now.

 

I do feel great now but let's see what happens when I revert back to my old similar but more rapid protocol. I do plan to keep it otherwise the same but cycle for adaption this time around though, not necessarily for fission/fusion.

Edited by Nate-2004, 03 January 2018 - 06:47 PM.

[Turnbuckle]

 

I don't know for sure that there is any real way to get QC on dysfunctional or mutated mitochondria but it's one of the major targets in aging research, one in which SENS proposes a solution of storing "backup copies" (see point 3) in the cell nucleus, rather than attempting to weed out the baddies so to speak. I think this is something I'll have to read more about but perhaps there's just not enough information to draw any actionable conclusions right now.

 

 

Cells have backup copies already, as the average cell has some thousand mitochondria and each has several mtDNA loops. Combine that with fusion that exchanges proteins, and even highly damaged mitochondria can keep on trucking. At least until they reach the point that every mtDNA is damaged, and then there is no saving the cell. So it's important to eliminate defective mtDNA before that point. You can do that with fission where mitochondria are divided up into minimal size with a single loop of mtDNA. If that loop has a defective gene, the membrane potential drops and the mitochondrion is marked for lysosomal digestion and cannot re-fuse with other mitochondria. Then when you restock the cellular population of mitochondria with fusion/biogenesis, the population is less damaged than before. Do multiple cycles and you can restore a more youthful state in a step-wise fashion. After that you could go into a maintenance mode of just topping off your NAD+ level until defective mitochondria build up again. (And you might find the high fission state useful for exercise at any time.)

 

As for the speculative idea of modifying human DNA to put the 37 mito genes into the nucleus, evolution has already moved most human mtDNA there except for that handful, and it is not theoretically impossible to move it all as a few organisms have managed to do this. But doing that in our lifetime seems unlikely. 

Edited by Turnbuckle, 03 January 2018 - 08:11 PM.

[Turnbuckle]

Protocol without NAD+ (experimental)

 

Rationale: Taking NAD+ precursors for long periods could produce a dependent state where mitochondria become addicted to the exogenous supplements due to the buildup of defective genes related to the internal production of NAD. These defects could thus be hidden from the cells QC by the superabundance of NAD. Alternating the fission routine without supplemental NAD should avoid this problem. 

 

Fission/mitophagy (before bed, 1-3 days)

Apigenin (fission) — 100mg

Fisetin (Sirt1 activator) — 100mg

AMPK activators (Life Extension) — 1g

 

(to convert this to the NAD version, add 2g each of nicotinamide and ribose and delete the apigenin)

 

Fusion/Biogenesis

Stearic acid (fusion) — 5g

Leucine (biogenesis) — 5g

PQQ (biogenesis) — 20mg

Hydroxytyrosol (biogenesis & antioxidant) — 25mg

Vitamin B complex (commercial mix)

 

or, if returning to fission the next day-- 

 

Fusion/Biogenesis 

Broccoli sprout extract with sulforaphane (fusion) — .5-1g

Leucine (biogenesis) — 5g

PQQ (biogenesis) — 20mg

Hydroxytyrosol (biogenesis & antioxidant) — 25mg

Vitamin B complex (commercial mix)

 

 

 

References

 

The dynamic regulation of NAD metabolism in mitochondria

 

A growing body of evidence also suggests that mitochondria have their own NAD biosynthetic machinery which appears to play an important role in maintaining the mitochondrial NAD pool, in response to environmental and nutritional stresses. Although the details still remain unclear, how mitochondria maintain their NAD pool by their own NAD biosynthetic machinery and/or through the communication with nuclear/cytoplasmic NAD biosynthetic pathways has recently become a focus of intensive investigation.

 

https://www.ncbi.nlm...les/PMC3683958/

 

 

 

The Black Queen Hypothesis: Evolution of Dependencies through Adaptive Gene Loss

 

Reductive genomic evolution, driven by genetic drift, is common in endosymbiotic bacteria [such as mitochondria]. .. Gene loss can provide a selective advantage by conserving an organism’s [mitochondria in this case] limiting resources, provided the gene’s function is dispensable. Many vital genetic functions are leaky, thereby unavoidably producing public goods that are available to the entire community. [Such as the cell's NAD pool.] Such leaky functions are thus dispensable for individuals, provided they are not lost entirely from the community. [This hypothesis] predicts that the loss of a costly, leaky function [the mitochondria's own production of NAD] is selectively favored at the individual level and will proceed until the production of public goods is just sufficient to support the equilibrium community...

 

https://www.ncbi.nlm...les/PMC3315703/

 

 

 

Edited by Turnbuckle, 06 January 2018 - 12:36 PM.

[Nate-2004]

Apigenin has a half life of 92 hrs roughly. To get 100g apigenin you'd need about 300g parsley (~30mg / 100g), 3 or more celery sticks and possibly a glass of chamomile tea or three. 100g parsley is approximately 20 sprigs.  I put this into my CD38 inhibiting smoothies with celery. I have chamomile that night as well.

 

For 100mg apigenin it would take 26 days to eliminate it from your system.

 

Apigenin is said to inhibit CD38, a NAD+ consumer and possibly to blame for NAD+ deficiency as CD38 rises as we age (possibly due to SASP).

 

The real upstream solution, if it *is* SASP, would be the clearance or revival of senescent cells.  Novotoclax, Dasatinib, FOXO4-DRI, HSP90 inhibitors are on the horizon.

 

I think Turnbuckle's hypothesis is testable and worthy of exploration but I am not quite sure this new protocol will do anything as far as induced fission goes. You would have to prove somehow that by inhibiting CD38 with Apigenin the resulting increase in NAD+ is what fissions mitochondria. Hopefully we know more in 5 years about whether exogenous NAM+R or NR helps or hurts in the long run.

Edited by Nate-2004, 06 January 2018 - 03:59 PM.

[Turnbuckle]

Apigenin has a half life of 92 hrs roughly. To get 100g apigenin you'd need about 300g parsley (~30mg / 100g), 3 or more celery sticks and possibly a glass of chamomile tea or three. 100g parsley is approximately 20 sprigs.  I put this into my CD38 inhibiting smoothies with celery. I have chamomile that night as well.

 

For 100mg apigenin it would take 26 days to eliminate it from your system.

 

Apigenin is said to inhibit CD38, a NAD+ consumer and possibly to blame for NAD+ deficiency as CD38 rises as we age (possibly due to SASP).

 

The real upstream solution, if it *is* SASP, would be the clearance or revival of senescent cells.  Novotoclax, Dasatinib, FOXO4-DRI, HSP90 inhibitors are on the horizon.

 

I think Turnbuckle's hypothesis is testable and worthy of exploration but I am not quite sure this new protocol will do anything as far as induced fission goes. You would have to prove somehow that by inhibiting CD38 with Apigenin the resulting increase in NAD+ is what fissions mitochondria. Hopefully we know more in 5 years about whether exogenous NAM+R or NR helps or hurts in the long run.

 

 

Good points, Nate. The dose of apigenin could probably be reduced from the 100mg I posed above and only taken on the first day of a several day fission cycle. You don't need to get it from foods as 50mg caps are available. I've used 50mg before with the NAD+ protocol with no apparent problem, and 100mg seems not out of line. Apigenin is known to induce fission--

 

Apigenin (10 mM) decreased the number of cells undergoing mitochondrial fusion and induced mitochondrial fragmentation after 24 h.

https://www.ncbi.nlm...pubmed/25909344

 

 

 

This same paper talks of apigenin inhibiting quality control, but only insofar as it inhibits fusion, as they recognize that both fission and fusion are required. We of course are only using it as a temporary measure to enhance quality control and get rid of mitochondria that have become dependent on exogenous NAD.

Edited by Turnbuckle, 06 January 2018 - 04:42 PM.

[zorba990]

Protocol without NAD+ (experimental)

Rationale: Taking NAD+ precursors for long periods could produce a dependent state where mitochondria become addicted to the exogenous supplements due to the buildup of defective genes related to the internal production of NAD. These defects could thus be hidden from the cells QC by the superabundance of NAD. Alternating the fission routine without supplemental NAD should avoid this problem.

Fission/mitophagy (before bed, 1-3 days)
Apigenin (fission) — 100mg
Fisetin (Sirt1 activator) — 100mg
AMPK activators (Life Extension) — 1g

(to convert this to the NAD version, add 2g each of nicotinamide and ribose and delete the apigenin)

Fusion/Biogenesis
Stearic acid (fusion) — 5g
Leucine (biogenesis) — 5g
PQQ (biogenesis) — 20mg
Hydroxytyrosol (biogenesis & antioxidant) — 25mg
Vitamin B complex (commercial mix)

or, if returning to fission the next day--

Fusion/Biogenesis
Broccoli sprout extract with sulforaphane (fusion) — .5-1g
Leucine (biogenesis) — 5g
PQQ (biogenesis) — 20mg
Hydroxytyrosol (biogenesis & antioxidant) — 25mg
Vitamin B complex (commercial mix)



References

The dynamic regulation of NAD metabolism in mitochondria

A growing body of evidence also suggests that mitochondria have their own NAD biosynthetic machinery which appears to play an important role in maintaining the mitochondrial NAD pool, in response to environmental and nutritional stresses. Although the details still remain unclear, how mitochondria maintain their NAD pool by their own NAD biosynthetic machinery and/or through the communication with nuclear/cytoplasmic NAD biosynthetic pathways has recently become a focus of intensive investigation.

https://www.ncbi.nlm...les/PMC3683958/

The Black Queen Hypothesis: Evolution of Dependencies through Adaptive Gene Loss

Reductive genomic evolution, driven by genetic drift, is common in endosymbiotic bacteria [such as mitochondria]. .. Gene loss can provide a selective advantage by conserving an organism’s [mitochondria in this case] limiting resources, provided the gene’s function is dispensable. Many vital genetic functions are leaky, thereby unavoidably producing public goods that are available to the entire community. [Such as the cell's NAD pool.] Such leaky functions are thus dispensable for individuals, provided they are not lost entirely from the community. [This hypothesis] predicts that the loss of a costly, leaky function [the mitochondria's own production of NAD] is selectively favored at the individual level and will proceed until the production of public goods is just sufficient to support the equilibrium community...

https://www.ncbi.nlm...les/PMC3315703/

Thanks for the new info.

LEF recently changed their AMPK formula. Do you still support using the new formula with hesperidin?

Serving Size 1 vegetarian tablet
Amount Per Serving
Calcium (as calcium carbonate)
130 mg
Hesperidin [from orange extract (fruit)]
500 mg
ActivAMP® gynostemma extract (leaf)
450 mg
Other ingredients: microcrystalline cellulose, stearic acid, croscarmellose sodium, hydroxypropyl cellulose, aqueous film coating (hypromellose, glycerin, purified water), silica, vegetable stearate.

[Turnbuckle]

 

LEF recently changed their AMPK formula. Do you still support using the new formula with hesperidin?

 

 

I can't tell you why they changed it or whether it might be better. Gynostemma from Immortalitea gets better ratings at Amazon, and that will probably be sufficient.

 

A note on post 716 above. I copy pasted a quote from a Apigenin paper, and μM changed to mM. The actual concentration was 10 micromolar.

Edited by Turnbuckle, 06 January 2018 - 10:52 PM.

[Turnbuckle]

I’ve found that the following protocol works at least as well as the previous ones, while reducing the potentially negative effects of too much ribose and the potential for mitochondria becoming dependent on NAD supplements—

 

 

Five-day protocol with reduced NAD and ribose

 

 

Day 1: Fission/mitophagy

 

Morning

Nicotinamide (NAD+) — 2g

Ribose (NAD+) — 2g

Jiaogulan leaf (AMPK activator) — 1g

Apigenin (fission, has a long half-life) — 100mg

Fisetin (Sirt1 activator) — 100mg

 

Evening

Jiaogulan leaf — 500mg

Fisetin — 100 mg

 

Day 2-3: Fission/mitophagy

 

Morning & Evening

Jiaogulan leaf  — 500mg

Fisetin — 100 mg

 

Day 4: Fusion/biogenesis

 

Morning

Stearic acid (fusion) — 5g

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

Edited by Turnbuckle, 12 January 2018 - 12:00 AM.

[stephen_b]

I got a lipid profile back, and this protocol seems to lower cholesterol. My total cholesterol went from 211 to 178, and LDL was reduced from 145 to 114 mg/dL (LDL calculated). HDL was unchanged at around 40.

 

That would seem to provide a bit of anecdotal evidence that niacinamide  + riboside is more effective than niacinamide by itself. I was taking 1.5 g nicotinamide and about twice that in riboside.