179 replies to this topic

[Turnbuckle]

Object: A protocol to stimulate an innate cellular mito quality control routine to destroy unhealthy mitochondria, followed by stimulating mitochondrial biogenesis and increasing ATP production.

 

The experimental protocol:

 

(1)   High dose nicotinamide (one dose, 1-2 grams)

(2)   Induction time (1-2 days? Defective mtDNA and mitochondria are recycled)

(3)   PQQ (three doses, 10 mg per dose every 6 hours)

(4)   Induction time (1-2 days? New mtDNA and mitochondria are created)

(5)   C60 (one dose, 3-4 mg)

(6)   Time (1-4 weeks? Functioning of all mitochondria is improved)

(7)   Repeat

 

Background: PQQ and C60 are used for higher energy and possibly longer life. The mechanism for C60 is still unknown, but user experiences suggests enhanced ATP production is one feature. PQQ is known to stimulate biogenesis of mitochondria, and thus also more ATP production. But few ask what happens when the cells are full of mitochondria and if the cell is capable of eliminating mitochondria (mitophagy) with any sort of quality control. Until very recently, this was unknown.

 

 

Rationale for the protocol:

 

(1) Certain substances are able to dramatically increase mitophagy, by which cells eliminate poor functioning mitochondria. Cells use the ratio of NAD+ to NADH (the oxidized and reduced forms of nicotinamide adenine dinucleotide) as a signal to begin mitophagy. Precursors to NAD such as nicotinamide (and also niacin and nicotinamide riboside) can increase this ratio and dramatically increase mitophagy—

 

Reference: Nicotinamide-induced mitophagy: event mediated by high NAD+/NADH ratio and SIRT1 protein activation.

 

(2) It appears that mitophagy occurs very fast, within one to two days. The unwanted mitochondria are destroyed by lysosomes, and the speed of the process is likely related to the age of the individual. Unfortunately, lysosomes collect indigestible garbage (lipofuscin, for instance, and possibly C60 as well), which slow things down.

 

Reference: The mitochondrial-lysosomal axis theory of aging

 

(3-4) PQQ (Pyrroloquinoline quinone) stimulates mitochondrial biogenesis, and this is rather rapid as well. It appears to be proportional to dose and time.

 

Reference: Pyrroloquinoline Quinone Stimulates Mitochondrial Biogenesis . . .  See Fig 1, A & C.

 

(5-6) If C60 improves all mitochondria (speculative) then that will reduce the ability of the cell machinery to distinguish good from bad mitochondria, as this is likely based on mitochondrial membrane potential. Thus, to be on the safe side, sufficient time should be allowed before the next nicotinamide dose. Certainly they should not be taken at the same time. Some years back I tried a large dose of niacin along with C60, and it took a week before I felt normal again. On the other hand, I don’t see why steps (3-4) and (5-6) couldn’t be combined.

 

 

 

[MissMaggie]

I really, really like the way you think, Turnbuckle. Have you attempted a version of this proposed protocol yet, or waiting for comments / warnings first?

[Turnbuckle]

I have given it one shot, Maggie, though the protocol was slightly different from what I suggested above. The first dose was 1.5 grams nicotinamide plus 20 mg elemental lithium from lithium orotate. I've had a lot of experience with niacin, taking it for years at the 3 gram level, but nicotinamide proved to be different. Within a few hours I felt a bit like I had the flu, then fell into a deep sleep for hours in the middle of the day. Which wouldn't be surprising if it were really knocking off mitochondria. The next day I took 20 mg PQQ, and then several hours later took a 3 mg dose of C60 in MCT oil. By the third day I had an unusual amount of energy. To get the best results, I expect this would have to be repeated a number of times, as it seems unlikely the cellular mechanism is capable of eliminating all bad mtDNA in one pass, as each mitochondrion has several copies and are constantly exchanging them through fission and fusion, so it's a game of percentages.

[MissMaggie]

makes sense, and signs very promising. fortunate that nicotinamide is very cheap, and mega doses seem to be relatively well tolerated (though toxic 3mg or more?). I've been on Mitoq for about 5 months, PQQ for 4-ish, and c60oo for almost 2 months. if one were to add Mitoq to this protocol, where would you place it? with the PQQ or the c60?

[Turnbuckle]

makes sense, and signs very promising. fortunate that nicotinamide is very cheap, and mega doses seem to be relatively well tolerated (though toxic 3mg or more?). I've been on Mitoq for about 5 months, PQQ for 4-ish, and c60oo for almost 2 months. if one were to add Mitoq to this protocol, where would you place it? with the PQQ or the c60?

 

 

I take mitochondrial supplements on a regular basis--CoQ10, ALCAR, ALA, GSH--but I would avoid all such supplements during steps (1-2), since antioxidants can inhibit autophagy.

 

Supplements that enhance mito fission (or block fusion) should be helpful during steps (1-2), since mitochondria must be reduced to their smallest size to allow lyosomes to engulf them. The actual process involves surrounding the dysfunctional mitochondrion with a double-membrane vesicle and delivering it to a lysosome for hydrolytic degradation. Oversized, dysfunctional mitochondria of the elderly can thus escape destruction and clog up the cell.

 

As for supplements that might enhance fission--

 

 

These data suggest that FAs, particularly PA, are an external factor altering mitochondrial dynamics and shifting the balance toward fission in muscle cells.

 

http://mcb.asm.org/c...t/32/2/309.full

 

 

The PA this refers to is palmitate, which is believed to increase mtROS, the action of which can be blocked by mito antioxidants. So while it's not clear that palmitate will help with steps (1-2), it seems clear that mito antioxidants will hurt. It also suggests that the daily use of mito antioxidants could result in a growing load of dysfunctional mitochondria.

Edited by Turnbuckle, 11 April 2016 - 11:01 AM.

[Rocket]

What about sr9009 reportedly growing new mitochondria?

[pamojja]

The first dose was 1.5 grams nicotinamide plus 20 mg elemental lithium from lithium orotate. I've had a lot of experience with niacin, taking it for years at the 3 gram level, but nicotinamide proved to be different. Within a few hours I felt a bit like I had the flu, then fell into a deep sleep for hours in the middle of the day. Which wouldn't be surprising if it were really knocking off mitochondria.

 

Do you have any theory why nicotinamide showed that different from regular niacin?

[Turnbuckle]

 

The first dose was 1.5 grams nicotinamide plus 20 mg elemental lithium from lithium orotate. I've had a lot of experience with niacin, taking it for years at the 3 gram level, but nicotinamide proved to be different. Within a few hours I felt a bit like I had the flu, then fell into a deep sleep for hours in the middle of the day. Which wouldn't be surprising if it were really knocking off mitochondria.

 

Do you have any theory why nicotinamide showed that different from regular niacin?

 

 

 

Nicotinic acid (niacin) and its amide nicotinamide (niacinamide) are said to both convert to NAD, but it's not clear how they compare in efficiency. Certainly they don't have the same biological effect. Niacin produces pronounced flushing and modifies cholesterol profiles, while nicotinamide does neither, and the biological half life of nicotinic acid is around one hour while that for the amide is much longer--4-6 hours. So for the half life alone, niacinamide seems much better for the protocol of the OP. Both have been recommended for sleep, though I've seen some say that niacin worked while niacinamide didn't, so it seems there's a lot of individual variation.

[Turnbuckle]

What about sr9009 reportedly growing new mitochondria?

 

 

Certainly. I was just using supplements commonly available and that could be taken orally as examples. The protocol is simply a recognition that the cell has an inbuilt process of mitochondria quality control--fission and fusion, mitophagy and biogenesis--and supplements should be taken in a certain sequence so as to work with the cell and not screw up the process. 

Edited by Turnbuckle, 12 April 2016 - 12:30 PM.

[Turnbuckle]

Another trial of the OP hypothesis—

 

(1) 2 grams nicotinamide + 20 mg elemental lithium from lithium orotate. This I took late in the evening to avoid falling asleep during the day.

 

(2) Next day I took 40 mg PQQ throughout the day, plus 3 mg C60 (mix #8) that morning, plus my regular regime of supplements as indicated above in post #5. I felt fine, but running was substantially off, as expected.

 

(3) Second day, supplements as indicated in (2). Running was back 100% and I felt a good deal of muscle tone. Gym was more than 100% and I maxed out the weights of one machine I hadn’t gone past 2/3rds in several years.

 

Conclusion: The overall effects were good, but perhaps less than the first round.  This is as I would expect from the way cells handle mito quality control. The process is never 100% efficient, and thus rounds of treatment should eliminate a declining number of defective mtDNA strands (the ultimate source of defective mitochondria). My plan is to allow 2-4 weeks before I try this again.

 

Edited by Turnbuckle, 19 April 2016 - 11:28 AM.

[Empiricus]

(1) I took 1 gram nicotinamide riboside (NR) with 10 mg elemental lithium before bed.  Though I didn't sleep very well and felt some cold symptoms, by the following afternoon I felt rather rejuvenated.  

 

(2) Thirty-six hours later, I was feeling a rather run down again.  That's when I took first of 4 doses of PQQ at 20 mg per dose every 6 hours.  The next day I woke up with ringing in ears.  I feel very low energy and lethargic for the next couple days.  

 

Four days later I took 100 mg of NR spaced 12 hours apart and that perked me up both times.  I still hear ringing when I wake up, but the sound goes away after a while or at any rate I don't notice it.  I read somewhere that NR is a treatment for tinnitus, but I hadn't realized too much PQQ could set it off.  

 

Funny thing is before this experiment I had concluded NR did nothing (I just used some leftover tablets in a bottle) and yet I swore by PQQ. Now I am seeing real benefits of NR and becoming wary of PQQ. 

 

I'm taking a break from c60oo, so nothing to report on that.  

[momof08]

Hi,

re:  your protocol as outlined on this page.  Would you share your source for your supplements and your reason for purchasing that particular brand?  I'm a newbie and need all the info I can get--I'm also "old" and need all the help I can gather in order to stay where I am or improve!  Thanks,  

[Vitalist]

What about sr9009 reportedly growing new mitochondria?

 

I'm surprised to see that in several places on this forum people have admitted taking or are considering taking SR9009 since a quick search will turn up comments from the drug's lead developer, Professor Burris, such as this:

emailed professor burris. This is what he said.
"I just recently found out that someone is selling SR9009 in a manner that is "supported" for human use in bodybuilding. I agree with your comments. The drug does in fact alter the circadian rhythm (in mice) and we would need to assume in humans – and we don't know if it is beneficial or detrimental at this point. SR9009 was designed as a "tool" molecule to determine if we should pursue making more drug like compounds that could be used to treat disease. Of course, the data is supportive of going forward – and we are designing better compounds – but the issue is that SR9009 has several issues that make it unsuitable for human use. Firstly, it has no oral bioavailability. I know the company selling this is indicating taking it orally is ok – but it doesn't even get into the blood. At this point that is probably not a bad thing since the drug has not been evaluated in appropriate toxicology assays to determine if there are issues like inducing disease (cancer as you indicate or others). There is going to be exposure to the gut and that is not necessarily good since we don't know the effects as of yet. SR9009 has some functional groups that are known to have potential toxicology liabilities and it would never be developed as a drug. It was ok as a "tool" to figure out if we should continue to spend money and effort to get better drugs – but we had to design these "bad" functional groups out since they are know to have toxic effects in humans. So bottom line – I would never recommend using this compound at this point. That being said – we are still working on improved compounds with one of the potential uses being sarcopenia – loss of muscle and strength due to aging."

→ source (external link)

 

I guess the good news is that since SR9009 has almost zero oral bioavailability, it probably won't kill you right away. You're probably just wasting your money.

 

 

[MissMaggie]

Have been reading more about the difference between Niacin and Nicotinamide, came across the research of the rather controversial Dr Abram Hoffer.

http://www.doctoryou...fer_cardio.html

I think a 90 percent reduction in mortality compared to placebo from extended supplementation is pretty damn impressive. He claims nicotinamide (aka niacinamide) doesn't have the same effects, and it's clear it doesn't in terms of cholesterol, but might there be some merit to using both forms perhaps, given the impressive longevity effects claimed of niacin specifically? And if so, any opinions on taking them together or one after the other? Niacin seems to often cause sleeplessness, so maybe Niacin in the morning and Nicotinamide before bed? I've also seen a number of references to continued Niacin use over long periods eventually becoming ineffective, so the theory of cycling rather than taking together with mitochondrial biogenesis supplements would be borne out perhaps in light of these kinds of reports, and would maybe not reduce in effectiveness long term. And even an occasional cycle as opposed to constant eg one week per month or less often, may be sufficient to spring clean the mitochondria, perhaps with increasing frequency at greater age?

[Turnbuckle]

Have been reading more about the difference between Niacin and Nicotinamide, came across the research of the rather controversial Dr Abram Hoffer.

http://www.doctoryou...fer_cardio.html

I think a 90 percent reduction in mortality compared to placebo from extended supplementation is pretty damn impressive. 

 

 

 

Hoffer is referring to unpublished work by this fellow E. Boyle that had no controls and no way to judge the claims made. He then refers to different work that may or may not have been associated with Boyle in some way, but appears to have been properly done with large trial groups and a control group. He reports the results as if they were good, and maybe they were, but they weren't anything like Boyle's results--

 

 

The 1985 follow-up study showed no significant differences in mortality between those treatment groups which had been discontinued and placebo or Clofibrate. However, to the investigator's surprise, the niacin group fared much better. The cumulative percentage of deaths for all causes was 58.4%, 56.8%, 55.9%, 56.9% and 50.6% for low dose estrogens, high dose estrogens, Clofibrate, dextrothyroxine, placebo and niacin, respectively. 

 

The mortality in the niacin group was 11 percent lower than in the placebo group (P = 0.002). The mortality benefit from niacin was present in each major category or cause of death: coronary, other cardiovascular, cancer and others. Analysis of life table curves comparing niacin against placebo showed the niacin patients lived two years longer. With an average followup of fourteen years, there were 70 fewer deaths in the niacin group than would have been expected from the mortality in the placebo group. Patients with cholesterol levels higher than 240 mg per 100 mL benefited more than those with lower levels.

 

 

 

It's impossible to say much about this except Hoffer is rather sloppy. He gives five results when he should have six--

From Clinicaltrials.gov, these groups were:

 

2.5 mg/day of conjugated estrogens

5.0 mg/day of conjugated estrogens

1.8 gm/day of clofibrate

6.0 mg/day of dextrothyroxine sodium

3.0 gm/day of niacin

3.8 gm/day of lactose placebo.

 

No results were posted on the clinical trials site, but I found them on a French site here, though these were results at 6.2 years rather than 14. The ratio of all cause mortality of niacin to placebo was 0.96. That's to say, at the follow up at 6.2 years, niacin lowered the all cause death rate by 4%. Above, Hoffer claims it was 11% at 14 years. That might be true, but it's far from the 90% mortality reduction for "patients who remained on niacin for ten years and received individual attention" that Hoffer begins his story with, so he's being less than honest in his presentation. He's bolstering an uncontrolled study with a controlled one that had far less impressive results. To be honest, he should have given us actual mortality rates and ages for Boyle's patients so we could decide for ourselves how this 90% reduction claim fared when compared to this later controlled study.

Edited by Turnbuckle, 24 April 2016 - 11:10 AM.

[MoreNowAgain]

Where are you getting your PQQ?

[MissMaggie]

Thanks Turnbuckle, I did say he was controversial

Good points, but I was rather more interested in discussion on the pros and cons of niacin vs nicotinamide. I've also now read a bit more about nicotinamide riboside, which is under patents and thus bloody expensive, but looks to have some very promising features and trying to work out how much more bioavailable / useful it might be. Any comments on niacin vs ncotinamide vs NR for this protocol?

[Turnbuckle]

Thanks Turnbuckle, I did say he was controversial

Good points, but I was rather more interested in discussion on the pros and cons of niacin vs nicotinamide. I've also now read a bit more about nicotinamide riboside, which is under patents and thus bloody expensive, but looks to have some very promising features and trying to work out how much more bioavailable / useful it might be. Any comments on niacin vs ncotinamide vs NR for this protocol?

 

 

Cold dosing niacinamide at the gram + level is probably not a problem for most people. But niacin is a different animal and the dose should be worked up to gradually. Even assuming it is as effective as niacinamide for increasing mitophagy, there's the question of how continuous treatment would effect the NAD+/NADH ratio referred to in the OP. One might expect homeostasis to result in a less dramatic effect than a single large dose. As for NR, I don't know, but it certainly looks interesting--

 

Treatment with the NAD+ precursor nicotinamide riboside (NR) induced the mitochondrial unfolded protein response (UPRmt) and synthesis of prohibitin proteins, and this rejuvenated MuSCs in aged mice. NR also prevented MuSC senescence in the Mdx mouse model of muscular dystrophy. We furthermore demonstrate that NR delays senescence of neural SCs (NSCs) and melanocyte SCs (McSCs), and increased mouse lifespan.

 

http://www.ncbi.nlm....pubmed/27127236

 

[MissMaggie]

Thanks! I have done more reading on Longecity about NR and may have possible found some useful info...

I don't think it's possible to quote from different threads, but the below is from BryanS from Nov15:

"Niacin still has a longer road to get to NAD than the other 2 "available precursors" and Niacin can tax the liver at therapeutic doses as it dedicates resources to build upon this molecule.

The Gpr109a receptor is the root of the flush and over time this can be desensitized. I could given time begin to tolerate the flush. Still for the population as a whole getting people to embrace the flush without giving up the daily regiment is difficult. Obviously as seen on the Longecity forums there are very dedicated Niacin users. The effects with the Gpr109a receptor have been associated with good lipid (cholesterol) management and this has been the draw for many but now there seems to be a change in the general consensus as to where these benefits arise. Just raising NAD levels may be the key and the SIRT1 and SIRT3 activation that results may be where this lipid benefit arises.

So if this is truly the case NR is the shortest route to NAD and SIRT1 and SIRT3 are both elevated as a primary effect and we also find the liver is not over taxed in the process. On the other hand cheap Nicotinamide (NAM) will also raise NAD levels and with this more SIRT3 is eventually produced but as we've read SIRT3 has a Nicotinamide binding pocket to inhibit activity so the end result is likely not as intense.

So I tend of look at all this thru the lens of nutrient sensors as it relates to reproduction and periods of famine. As we look at the Sirtuins they appear to modulate mitochondrial function to match nutrient supply. NR has the effect of a Calorie Restriction (CR) mimic and the cell responds by ramping up its vital maintenance process's protecting the cell until times of plenty. On the other hand Nicotinamide tells the cell I've got it good (nutritionally speaking) and in that world its not as imperative to extend the cells existence longer than normal for the reproduction of the organism.

In reality bumping up NAD with NA or NR will also eventually raise levels of NAM. We naturally swing between these states. As (NAM) levels increase eventually they begin to inhibit the very same (CR) benefits we seek. So its not just a black and white picture and these feedback loops are at work all thru the day and night. These nutrient swings are all tied to our circadian rhythm. So from this perspective I think we can help enhance each side of this cycle by supplementing with NR by day and using (NAM) Nicotinamide before bed.

Even as far back as 1976 the sedative effects of nicotinamide were being documented and this fits with the idea that its use helps reenforce our nighttime cycle. http://www.ncbi.nlm....v/pubmed/133816 Also from a nutritional standpoint the body doesn't need to gear up for the chase to feed itself if its being told nicotinamide levels are high. We can also find many other references to the use of nicotinamide in cases of schizophrenia to calm patients. I think the clinical observations are tied to a fundamental feedback loop telling the cell all is well, its time to relax.

So if this is the case and I think there is a lot of evidence to support this idea, we can enhance our natural cycle by timing what we take and what time we take it. JMHO"

As a side note I took 1g nicotinamide last night and feel really well rested today. I think I'll do one more night dose and then begin PQQ late tomorrow. I think I like the idea of following the circadian cycles here. But if Nicotinamide is encouraging mitophagy, and that is best achieved when at rest, what's happening with the NR? Is it actually doing something similar to PQQ? Maybe the protocol could instead be daytime / nightime rather than over several days?

[Turnbuckle]

But if Nicotinamide is encouraging mitophagy, and that is best achieved when at rest, what's happening with the NR? Is it actually doing something similar to PQQ? 

 

 

Good question. Does nicotinamide riboside act as a better nicotinamide with a shorter path to NAD+, or a combination of nicotinamide + PQQ? There is some evidence it's the latter. If so, then the PQQ step could be deleted, but I still believe the C60 step should be done subsequently so as not to interfere with quality control during the process. Also, NR should not be taken every day to avoid the potential homeostasis I mentioned above. In fact, perusing the Amazon postings on one NR product page, one user mentioned that NR was very good at first, but then faded, suggesting that a "homeostatic mechanisms tend to cause NAD+ increases to be transient." While that seems a strong possibility, but there are others, such as clogging the vacuoles with lipofuscin due to an excessive diet of mitochondria.

Edited by Turnbuckle, 01 May 2016 - 12:09 PM.

[Adam1]

Turnbuckle, I have been following various posts of yours with interest. Your protocol drew me in from lurking to reply. I find the parsimony appealing. Also I appreciate your conservative approach with a hormetic process such as mitophagy. However, I am wondering what makes you think the protocol is sufficient. Autophagy involves various pathways, and as numerous compounds (along with CR, methionine restriction etc.) can influence its different aspects, I am not certain what constitutes sufficient activation of mitophagy. Thoughts on a multi-targeted approach to activate autophagy with clearance of the resultant debris?

 

Also, I am not certain stimulation of mitophagy and mitogenesis/cell protection can be clearly demarcated. Compounds such as curcumin appear to activate autophagy while having opposite cell-protective functions. If autophagy is stimulated from various angles, it would likely involve overlapping and in some ways contradictory processes, but perhaps with a higher margin of safety??

[Turnbuckle]

Turnbuckle, I have been following various posts of yours with interest. Your protocol drew me in from lurking to reply. I find the parsimony appealing. Also I appreciate your conservative approach with a hormetic process such as mitophagy. However, I am wondering what makes you think the protocol is sufficient. Autophagy involves various pathways, and as numerous compounds (along with CR, methionine restriction etc.) can influence its different aspects, I am not certain what constitutes sufficient activation of mitophagy. Thoughts on a multi-targeted approach to activate autophagy with clearance of the resultant debris?

 

Also, I am not certain stimulation of mitophagy and mitogenesis/cell protection can be clearly demarcated. Compounds such as curcumin appear to activate autophagy while having opposite cell-protective functions. If autophagy is stimulated from various angles, it would likely involve overlapping and in some ways contradictory processes, but perhaps with a higher margin of safety??

 

 

All of your points are valid, Adam1. The process of mitophagy and mitochondrial biogenesis is natural and ongoing, but my thinking is to give it some help by dividing it into parts and then using the best supplements for each. The supplements I started with are cheap and available and not necessarily optimal, and it may even be better to combine them in some way. By contrast, a more expensive supplement like NR might stimulate both mitophagy and biogenesis at once. That it doesn't have a better reputation as a supplement may reflect on the way it's been marketed more than anything. People are taking it every day on the advice of the retailers, when once a week or once a month might be better. Because once defective mitochondria are eliminated, why keep stressing the cellular machinery? 

 

One can think of mitochondria as sheep on a farm (or bacteria in a vat), with each cell/farm having a herd of a thousand or so. Thus to maintain the best herd, sheep must be encouraged to breed, sick sheep must be culled, and the herd must be fed a nutritious diet. One could do all those things at once and all the time and end up with a herd that is far from optimal. For instance, look at the culling aspect. If all sheep act equally healthy from a diet rich in antioxidants and with ATP production jacked up by C60, then how does the cellular machinery know which ones to cull? So withholding supplements that can boost mito function before taking mitophagy boosting supplements would appear to make sense.

 

It's also true that larger mitochondria are more efficient for day to day use, but must be fissioned into smaller bodies for quality control and culling, so these are contradictory requirements.

 

You mentioned various pathways of autophagy, but mitophagy is a specialized type where only mitochondria are being culled. And exactly how hard is it? Not particularly, it seems, except in certain cases. For instance, if the lysosomes have become dysfunctional or full of indigestible cellular debris, then the trash can't be taken out. Or when large and dysfunctional mitochondria refuse to fission, then they become cellular junk, taking up space and leaking toxins. Eventually, the protocol will have to address these cases as well.

 

 

A possible factor limiting the effectiveness of mitochondial turnover is the enlargement of mitochondria which may reflect their impaired fission. Non-autophagocytosed mitochondria undergo further oxidative damage, resulting in decreasing energy production and increasing generation of reactive oxygen species. Damaged, enlarged and functionally disabled mitochondria gradually displace normal ones, which cannot replicate indefinitely because of limited cell volume. Although lipofuscin-loaded lysosomes continue to receive newly synthesized lysosomal enzymes, the pigment is undegradable. Therefore, advanced lipofuscin accumulation may greatly diminish lysosomal degradative capacity by preventing lysosomal enzymes from targeting to functional autophagosomes, further limiting mitochondrial recycling. This interrelated mitochondrial and lysosomal damage irreversibly leads to functional decay and death of postmitotic cells.

 

http://onlinelibrary...02.02869.x/full

 

[Adam1]

This is a useful model and is worth developing. Indeed, a regimen just to boost mitochondria may impede autophagy and thus may show initial benefit then hit a ceiling. On the other hand, an overly zealous autophagy regimen may result in excessive mitophagy, muscle wasting, frailty and fatigue. We want to cull organelles not the entire organism.

 

Mitochondria are a frequent target of autophagy, so the various pathways that network to result in autophagy apply to mitophagy, with additional pathways for directing the process to the mitochondria. Thus mitophagy may be even more multi-targeted than general autophagy. Ramping up the induction of autophagy could help fission of zombie mitos. Inducing autophagy in longer regimens may also benefit lysosomes that clogged up due to incomplete autophagy (insufficient steam to finish the job), although specific interventions might prove to activate the garbage pickup.

 

I really like the idea of a protocol with phases, although I believe the phases may necessarily end up overlapping. The timing of the phases is worth discussing. Circadian rhythms play a part, and over one night we typically have both anabolic and catabolic phases. This does not necessarily preclude longer cycles but suggests there is a place for very short cycles. Cycling over weeks with a brief intervention would be ideal and worth working on, but I don’t know of evidence to base it on. A minimalist approach is worth refining and testing to make sure it is sufficient.

[Turnbuckle]

Speculative method to kill zombie mitochondria—

 

Zombies are fat, lazy mitochondria that have ceased to function properly. They can be too big to fit into lysosomes and thus accumulate and eventually suffocate the cell.

 

An anti-zombie protocol might include—

 

(1)   Elimination of antioxidants that might hide them from quality control, and supplements that increase mito output that don’t also act as antioxidants. These would include trace minerals important to the citric acid cycle—zinc, copper, magnesium, molybdenum, manganese and others. Note that zinc is an antioxidant, but I suspect it will be of more help here than harm.

 

Next, mito fission and clearance should be encouraged—

 

(2)   Supplements that enhance the genes PINK1 and Parkin, which direct fission and clearance. Such supplements would be include cinnamon and sodium benzoate. Cinnamon alone should be sufficient, as it is metabolized to sodium benzoate.

 

Finally, enhancing the NAD+/NADH ratio and SIRT1 will set the quality control mechanism running—

 

(3)   For NAD+, lithium, niacin, nicotinamide and NR, and for SIRT1, resveratrol, tyrosol and hydroxytyrosol

 

 

References--

 

When normal mitophagic organelle elimination is suppressed by Parkin insufficiency, abnormal undead or zombie mitochondria accumulate and (as zombies will do) contaminate the normal mitochondrial population by fusing with normal organelles. Mitochondrial fusion that is ordinarily protective, therefore, becomes the mechanism for a general contagion of mitochondrial dysfunction. Interrupting mitochondrial fusion prevents contamination of functionally normal mitochondria by virulent zombie mitochondria, sequestering abnormal mitochondria that can then be removed by alternate, albeit almost certainly less efficient, elimination pathways.

 

Inhibiting mitochondrial fusion to contain mitochondrial contagion induced by mitophagic dysfunction may have applications in human diseases caused by defective mitochondrial quality control, most notably hereditary Parkinson disease induced by PINK1 and Parkin mutations.

 

http://www.ncbi.nlm....les/PMC4392818/

 

 

 

We have revisited the role of PINK1 and Parkin in mitochondrial dynamics, and explored its relation to the mitochondrial clearance program controlled by these proteins. We show that PINK1 and Parkin promote Drp1-dependent mitochondrial fission by mechanisms that are at least in part independent.

 

http://www.sciencedi...167488914001591

 

 

 

Mitochondrial Regulation by PINK1-Parkin Signaling

http://www.hindawi.c...rn/2012/926160/

 

Enhancing nucleotide metabolism protects against mitochondrial dysfunction and neurodegeneration in a PINK1 model of Parkinson’s disease

http://www.ncbi.nlm....les/PMC4199097/

 

Cinnamon Treatment Upregulates Neuroprotective Proteins Parkin and DJ-1 and Protects Dopaminergic Neurons in a Mouse Model of Parkinson’s Disease

http://www.ncbi.nlm....les/PMC4167597/

 

Sodium Benzoate, a Metabolite of Cinnamon and a Food Additive, Upregulates Neuroprotective Parkinson Disease Protein DJ-1 in Astrocytes and Neurons

http://www.ncbi.nlm....les/PMC3189510/

Edited by Turnbuckle, 04 May 2016 - 11:55 AM.

[Adam1]

That was helpful and interesting. Indeed PINK1 and Parkin are critical in mediating fission and mitophagy and are worth enhancing. Further mix ‘n match candidates for the autophagy portion of the protocol (mostly from Nixon, Ralph A. "The role of autophagy in neurodegenerative disease." Nature medicine 19.8 (2013): 983-997.):

 

TORC1 inhibition – curcumin

AMPK activation – lithium, trehalose, quercetin, catechins, naringen, berberine, curcumin

PP2A activation – palm oil for tocopherols, melatonin, forskolin, epigallocatechine gallate

Other induction targets: parthenolide in feverfew, fisetin, luteolin

 

Many phytochemicals can have oxidant or antioxidant effects depending on factors such as dose and what they encounter. Some compounds may both induce and inhibit autophagy. For example, berberine is also an inhibitor (Hundeshagen, Phillip, et al. "Concurrent detection of autolysosome formation and lysosomal degradation by flow cytometry in a high-content screen for inducers of autophagy." BMC biology 9.1 (2011): 38.) It appears specific antioxidants can have a direct role in mitophagy (Yang, Lei, Xuping Wang, and Xiaolan Yang. "Possible antioxidant mechanism of melanoidins extract from Shanxi aged vinegar in mitophagy-dependent and mitophagy-independent pathways." Journal of agricultural and food chemistry 62.34 (2014): 8616-8622.)

 

Ideally the protocol would be pared down and individualized to contain just the rate-limiting steps of mitochondrial turnover through mitophagy, biogenesis, dynamics of fusion/fission, and could also address selective protein quality control. I am interested in learning about ways of implementing this.

[Turnbuckle]

Zombie protocol trial—

 

(1)   Minerals as per post #24.

(2)   After one hour (arbitrary), 4 grams of cinnamon extract (Natrol) 

(3)   After another hour, 1.5 grams nicotinamide and 20 mg elemental lithium from lithium orotate, along with 50 mg hydroxytyrosol and 250 mg trans-resveratrol (+50 mg DHEA to avoid joint pain)

 

Results--

 

Day 0: Vague flu like symptoms

Day 1: Took 20 mg PQQ in one dose & 3 mg C60. Felt fine except for some muscle soreness as if I had worked out the day before. Smell and hearing seemed enhanced.  An hour later running was definitely off and I didn't feel energetic enough for the gym.

Day 2: Took 20 mg PQQ in one dose & 3 mg C60. Muscle soreness as on Day 1. Distance vision enhanced along with smell and hearing. Mentally, a slight nootropic effect. Running a bit better than normal, gym a bit worse.

Day 3: No PQQ or C60: No soreness. Senses and mental enhancement still there. Running better than normal.

 

Conclusion: There's no way for a self-experimenter to know if zombie mitochondria were affected, but there seemed to be some effect where ATP production seems to have dropped off and built back up over several days to a point above baseline. This is consistent with the elimination of defective mitochondria and their replacement. The effect on senses is not surprising as they are dependent on the functioning of mitochondria, however such effects are notoriously prone to subjective bias.

 

Edited by Turnbuckle, 07 May 2016 - 06:16 PM.

[sthira]

Have you tried prolonged fasting to possibly remove zombie mitochondria?

[Adam1]

I think even the short "fasting" during sleep would be potentially synergistic if the compounds are taken at night or before breakfast.

 

This personal trial is helpful to share. I experienced vague flu-like symptoms after a sharp increase in autophagy induction, so I think I know what you mean, Turnbuckle. However, just to rule one thing out, as you probably know cassia cinnamon contains coumarin which is a bit hepatotoxic, so in higher doses the safer Sri Lankan cinnamonum verum might be worth a try to compare side-effects. Also, rosemary might have a similar effect to cinnamon (Lin, Chia-Yuan, and Chia-Wen Tsai. "Carnosic acid protects SH-SY5Y cells against 6-hydroxydopamine-induced cell death through upregulation of parkin pathway." Neuropharmacology (2016).)

[Empiricus]

However, just to rule one thing out, as you probably know cassia cinnamon contains coumarin which is a bit hepatotoxic, so in higher doses the safer Sri Lankan cinnamonum verum might be worth a try to compare side-effects.

 

This article compares the two varieties: 

Little Bit of Spice for Health, but Which One? http://www.wsj.com/a...135502891970942

 

Here's a brand of ceylon cinnamon:

- Oregon's Wild Harvest, True Cinnamon

Here's a standardized extract of Indonesian Cinnamomum burmannii. The manufacturer uses an extraction process that supposedly eliminates the toxic compounds:

- Bluebonnet Nutrition, Cinnulin PF, Cinnamon Bark Extract

 

[Turnbuckle]

Many phytochemicals can have oxidant or antioxidant effects depending on factors such as dose and what they encounter. Some compounds may both induce and inhibit autophagy. 

 

My concern with anti-oxidants is only that they might interfere with the quality control process during the short period of the trial. 

 

Have you tried prolonged fasting to possibly remove zombie mitochondria?

 

 

No. I'm trying to do this the easy way.

 

However, just to rule one thing out, as you probably know cassia cinnamon contains coumarin which is a bit hepatotoxic...

 

 

I've used this particular cinnamon before with no such effect, so I don't believe it's the cinnamon alone. However, in my next trial I will try sodium benzoate instead.

Edited by Turnbuckle, 08 May 2016 - 10:31 AM.