2195 replies to this topic

[Turnbuckle]

I have been giving the high energy protocol of post #746 a go for the past couple of days. I notice right away a lower heart rate, both resting heart rate (near 54 bpm, down almost 10 bpm) and exercising heart rate during a 6 mile run.

 

While my heart rate was lower than normal for the same effort and I felt more energy, I still felt off. Maybe it was a higher blood pressure (I wasn't able to measure it). Perhaps the dose is too high? I might try cutting it in half and going out for a run again tomorrow.

 

I don't recommend anyone trying the high energy protocol without some way of measuring their BP. Automatic BP units are very cheap. The  Omron Bp710n 3 Series is available from Walmart and Target for $25.

[PAMPAGUY]

An update on Protocol #746.  Have been following the protocol for 4 weeks now and have had a significant improvement in energy and no side effects as in previous protocols.  Started out at 1.5g N+R + 5g Stearic Acid for 3 days.  It was too much for me, so I reduced it down to 1.5g first day, and 1gm 2nd and 3 day.  5 gm Stearic Acid daily.  Will slowly increase dosage up to 2 gm x 3 days as my body adjusts.  I have had no blood pressure problems at all and I monitor it daily.  Pulse rate goes up 5-10 bpm during protocol days. (normal resting HR is low 50's) I discontinue my Metformin for those 3 days, as it acts as a uncoupler.  Because of the lack of side effects especially on fission days of previous protocols, I look forward to my 3 days of treatment.  My energy level is equivalent to someone 50 years old, and I'm 71.  I do also take Rapamycin weekly, and it helped with my energy, but not to the same degree as this NAD precursor.  

 

I follow the ageing formula put forth in this paper.  http://www.oncotarge...3&path[]=57761 I take the complete formula and delete Metformin on treatment days.  FYI

[whileitravel]

 

I have been giving the high energy protocol of post #746 a go for the past couple of days. I notice right away a lower heart rate, both resting heart rate (near 54 bpm, down almost 10 bpm) and exercising heart rate during a 6 mile run.

 

While my heart rate was lower than normal for the same effort and I felt more energy, I still felt off. Maybe it was a higher blood pressure (I wasn't able to measure it). Perhaps the dose is too high? I might try cutting it in half and going out for a run again tomorrow.

 

I don't recommend anyone trying the high energy protocol without some way of measuring their BP. Automatic BP units are very cheap. The  Omron Bp710n 3 Series is available from Walmart and Target for $25.

 

 

I bought one and it was total crap. Completely false number. If you're  seriously monitoring your BP for health reasons, I wouldn't recommend it. I threw it away and just do it manually which isn't easy!

 

This apparently is a better option.

 

https://www.amazon.c...ds=panasonic bp

[PAMPAGUY]

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[PAMPAGUY]

Turnbuckle you mentioned in a previous post that the high energy protocol is for only 4 weeks.  I have done 4 weeks, (at a reduced dose) and am reluctant to give it up.  For me it has been superior to any other protocol.  I feel so good.  It would seem that the 3 on and 4 off schedule would give the mito sufficient time to do their own fission/fusion thing the natural way of producing there own NAD?  Perhaps with a week off every so often?

 

Would appreciate some feed back.

[Turnbuckle]

In post #746 I said I had tried it for 3 days. If you have done it for 4 weeks and its still working as well as it did at first, maybe it's all right. I've noticed on occasion with #746 (2 grams each of NAM and ribose) that the stearic acid doesn't always keep me in fusion, that it sometimes flips into fission after 8 hours (judging by the feel and exercise performance). If so, then it might provide a 2-in-1 fusion/fusion protocol, which would simplify things considerably. At any rate, taking breaks is likely wise.

[Nate-2004]

I came across this article here which is now confusing the crap out of me. These guys think that fission *causes* mitochondrial dysfunction if I'm reading it correctly. He also thinks high fat diets cause diabetes and insulin resistance, which is highly debatable.

 

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

 

Mitochondrial dysfunction in skeletal muscle has been implicated in the development of insulin resistance and type 2 diabetes. Considering the importance of mitochondrial dynamics in mitochondrial and cellular functions, we hypothesized that obesity and excess energy intake shift the balance of mitochondrial dynamics, further contributing to mitochondrial dysfunction and metabolic deterioration in skeletal muscle. First, we revealed that excess palmitate (PA), but not hyperglycemia, hyperinsulinemia, or elevated tumor necrosis factor alpha, induced mitochondrial fragmentation and increased mitochondrion-associated Drp1 and Fis1 in differentiated C2C12 muscle cells. This fragmentation was associated with increased oxidative stress, mitochondrial depolarization, loss of ATP production, and reduced insulin-stimulated glucose uptake. Both genetic and pharmacological inhibition of Drp1 attenuated PA-induced mitochondrial fragmentation, mitochondrial depolarization, and insulin resistance in C2C12 cells. Furthermore, we found smaller and shorter mitochondria and increased mitochondrial fission machinery in the skeletal muscle of mice with genetic obesity and those with diet-induced obesity. Inhibition of mitochondrial fission improved the muscle insulin signaling and systemic insulin sensitivity of obese mice. Our findings indicated that aberrant mitochondrial fission is causally associated with mitochondrial dysfunction and insulin resistance in skeletal muscle. Thus, disruption of mitochondrial dynamics may underlie the pathogenesis of muscle insulin resistance in obesity and type 2 diabetes.

[PAMPAGUY]

In post #746 I said I had tried it for 3 days. If you have done it for 4 weeks and its still working as well as it did at first, maybe it's all right. I've noticed on occasion with #746 (2 grams each of NAM and ribose) that the stearic acid doesn't always keep me in fusion, that it sometimes flips into fission after 8 hours (judging by the feel and exercise performance). If so, then it might provide a 2-in-1 fusion/fusion protocol, which would simplify things considerably. At any rate, taking breaks is likely wise.

Thanks for getting back to me so fast.  Of course, I'm using considerably less than you, but will work up in time.  I also feel that I still have some zombies to clear out.  Perhaps an experiment for you - up your Stearic Acid intake.

[Turnbuckle]

 

I came across this article here which is now confusing the crap out of me. These guys think that fission *causes* mitochondrial dysfunction if I'm reading it correctly. He also thinks high fat diets cause diabetes and insulin resistance, which is highly debatable.

 

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

 

 

 

Mitochondrial dysfunction in skeletal muscle has been implicated in the development of insulin resistance and type 2 diabetes. Considering the importance of mitochondrial dynamics in mitochondrial and cellular functions, we hypothesized that obesity and excess energy intake shift the balance of mitochondrial dynamics, further contributing to mitochondrial dysfunction and metabolic deterioration in skeletal muscle. First, we revealed that excess palmitate (PA), but not hyperglycemia, hyperinsulinemia, or elevated tumor necrosis factor alpha, induced mitochondrial fragmentation and increased mitochondrion-associated Drp1 and Fis1 in differentiated C2C12 muscle cells. This fragmentation was associated with increased oxidative stress, mitochondrial depolarization, loss of ATP production, and reduced insulin-stimulated glucose uptake. Both genetic and pharmacological inhibition of Drp1 attenuated PA-induced mitochondrial fragmentation, mitochondrial depolarization, and insulin resistance in C2C12 cells. Furthermore, we found smaller and shorter mitochondria and increased mitochondrial fission machinery in the skeletal muscle of mice with genetic obesity and those with diet-induced obesity. Inhibition of mitochondrial fission improved the muscle insulin signaling and systemic insulin sensitivity of obese mice. Our findings indicated that aberrant mitochondrial fission is causally associated with mitochondrial dysfunction and insulin resistance in skeletal muscle. Thus, disruption of mitochondrial dynamics may underlie the pathogenesis of muscle insulin resistance in obesity and type 2 diabetes.

 

 

You article speaks of the need for "balanced mitochondrial dynamics," ie fission and fusion. Fission alone will result in reduced mito mass, reduced ATP and increased ROS. And what about biogenesis during fission? It may not work too well with a single loop of mtDNA as good loops may get mistaken for bad ones and get recycled. The super-health state for the cell is fusion as that provides the most ATP, but fusion alone is also bad in the long run as defective mtDNA loops build up and you may not even realize it as fusion covers up the presence of bad loops.

Edited by Turnbuckle, 22 February 2018 - 07:57 PM.

[Black Fox]

Could anybody clarify if R-ALA is sth we should take in fusion days or fission? Cuz I got conflicting info, some papers say r-ALA is synergetic with NR ( therefore fission ) but if I'm not mistaken of the guys here are taking on fusion days ....

Could anybody throw some on light on this one?

Thanks in advance

[Turnbuckle]

Could anybody clarify if R-ALA is sth we should take in fusion days or fission? Cuz I got conflicting info, some papers say r-ALA is synergetic with NR ( therefore fission ) but if I'm not mistaken of the guys here are taking on fusion days ....

Could anybody throw some on light on this one?

Thanks in advance

 

 

Can you link to the papers you mentioned?

 

Most people taking NR aren't taking enough to get into deep fission. For example, if you are taking 2g each of N+R, that is equivalent to 4g of NR, while most taking NR are dosing at 250-500mg.

 

So if you are using N+R at these high levels to achieve fission, I would not use ALA. Instead I would take this with fusion as ALA & carnitine improve mito function, while with fission you're attempting to help cells find and destroy defective mitochondria (which exhibit low membrane potential). Supplements that improve membrane potential can improve cellular functioning, but also prevent otherwise defective mitochondria from being labeled as defective.

 

Neuronal mitochondrial amelioration by feeding acetyl-L-carnitine and lipoic acid to aged rats

 

Feeding old rats ALCAR and LA significantly reduced the number of severely damaged mitochondria (P = 0.02) and increased the number of intact mitochondria (P < 0.001) in the hippocampus. These results suggest that feeding ALCAR with LA may ameliorate age-associated mitochondrial ultrastructural decay and are consistent with previous studies showing improved brain function.

http://pubmedcentral...les/PMC2790425/

 

 

It's known that reducing lipoic acid synthesis reduces membrane potential, so you can expect that supplying lipoic acid would tend to raise it, thereby making it more difficult for cellular QC to identify defective mitochondria.

 

Downregulation of the LASY [Lipoic acid synthase] gene by RNA interference (RNAi) reduced endogenous levels of lipoic acid, and the activities of critical components of the antioxidant defense network, increasing oxidative stress. Treatment with exogenous lipoic acid compensated for some of these defects. RNAi-mediated downregulation of LASY induced a significant loss of mitochondrial membrane potential...

https://www.ncbi.nlm...les/PMC2646058/

 

 

One exception to this is my reversed protocol in post 746. Here N+R is combined with sufficient stearic acid to force mito fusion and produce a high energy state.

Edited by Turnbuckle, 23 February 2018 - 11:40 AM.

[BieraK]

I have not experiment with fission protocol for some time since I don't have stearic acid and Sulforaphane did not worked well for me, I developed some tendon pain doig N+R fission with weight training. Since January I'm doing some weights without high doses N+R and my tendons are ok, so another personal experience for the importance of cycling between fission and fussion and how some body areas have less mitos compared to others.
I'm for now only taking N+R in the 250 mg NR equivalent doses 3 times a day just for achieving a similar state compared to a high NAD+/NADH ratio. 

The next week I will do my second Fasting Mimicking Diet (FMD), the diet developed by Valter Long and team, the first time It was great I really liked it a lot, however I'm planning doing one or two days of fission, will report how it goes. The diet is like doing 5 days water fasting but without feeling all the pain and hunger, it also gives you some basic nutrients that are crucial for not harm the body while fasting.  

[Turnbuckle]

I have not experiment with fission protocol for some time since I don't have stearic acid and Sulforaphane did not worked well for me, I developed some tendon pain doig N+R fission with weight training. Since January I'm doing some weights without high doses N+R and my tendons are ok, so another personal experience for the importance of cycling between fission and fussion and how some body areas have less mitos compared to others.
I'm for now only taking N+R in the 250 mg NR equivalent doses 3 times a day just for achieving a similar state compared to a high NAD+/NADH ratio. 

The next week I will do my second Fasting Mimicking Diet (FMD), the diet developed by Valter Long and team, the first time It was great I really liked it a lot, however I'm planning doing one or two days of fission, will report how it goes. The diet is like doing 5 days water fasting but without feeling all the pain and hunger, it also gives you some basic nutrients that are crucial for not harm the body while fasting.  

 

 

I agree that stearic acid is better than sulforaphane, especially when combining N+R+fusion--and that is where you take your money off the table and get age regression. If you can't get stearic acid locally, try Amazon Global. They ship to 100 countries -- AmazonGlobal Export Countries. 

[BieraK]

Fortunately I can get stearic acid food grade, I tend to thin food grade is ok for human consumption.

Do you know what could be a equal dose of 250 NR but with Nicotinic Acid and D-Ribose instead of NAM?

I agree that Nicotinamide ande Niacin/Nicotinic Acid are different at the subjetive and body feeling, I Want to experiment with an equal dose of NA+R to 250 mg NR.

[Turnbuckle]

Fortunately I can get stearic acid food grade, I tend to thin food grade is ok for human consumption.

Do you know what could be a equal dose of 250 NR but with Nicotinic Acid and D-Ribose instead of NAM?

I agree that Nicotinamide ande Niacin/Nicotinic Acid are different at the subjetive and body feeling, I Want to experiment with an equal dose of NA+R to 250 mg NR.

 

 

Nicotinamide, nicotinic acid, and tryptophan all lead to NAD+, and ribose can be used with any or all of them at equal weights. The first is the closest chemically, has the fewest side effects, and has the longest half life. Here is a protocol I used at night containing tryptophan, nicotinamide and ribose (where I would now use 2.5 g ribose instead of 5). 

 

250 mg NR would be roughly equivalent to 125 mg of nicotinamide and 125 ribose. You could substitute equal parts of nicotinic acid for nicotinamide, but I can't tell you how the effects would compare.

Edited by Turnbuckle, 01 March 2018 - 12:04 PM.

[Turnbuckle]

Updated protocols--

 

 

Fission + mitophagy (2-3 days)

Nicotinamide (NAD+) — 2g

Ribose (NAD+) — 2g

AMPK activators (Life Extension) — 1g 

Fisetin (Sirt1 activator) — 100mg

 

Optional but recommended: Anaerobic exercise — 2-3 hours later. You should feel weaker, but more pumped up afterward. Then,

 

Fusion + Biogenesis (2-3 days)

Stearic acid (fusion) — 5g

Leucine (biogenesis) — 5g

PQQ (biogenesis) — 20mg

Hydroxytyrosol (biogenesis & antioxidant) — 25mg

Vitamin B complex (commercial mix)

Vitamin C — 1g

 

Fission then fusion removes bad mitochondria and replaces them. After doing this sequence a few months (depending on how bad your mitochondria were to begin with), the following reverse protocol can be used to achieve age regression by forcing fusion with high NAD+,

 

High energy — Fusion + high NAD+ (3 days or more, but not continuously)

Nicotinamide (NAD+) — 1g

Ribose (NAD+) — 1g

AMPK activators (Life Extension) — 1g 

Stearic acid (fusion) — 5-10g

 

Edited by Turnbuckle, 12 March 2018 - 05:57 PM.

[Andey]

 

Leucine (biogenesis) — 5g

 

 

Lysine? 

I suspect its a typo because 5g of Leucine is almost in every meal with some protein.

[Turnbuckle]

 

 

Leucine (biogenesis) — 5g

 

 

Lysine? 

I suspect its a typo because 5g of Leucine is almost in every meal with some protein.

 

 

This is no typo, and I suggest that leucine be used alone as including other amino acids can dilute or eliminate the desired effect. For the same reason stearic acid should be used alone and not in some mix with FAs, as some FAs can cause mito uncoupling.

 

Leucine Modulates Mitochondrial Biogenesis and SIRT1-AMPK Signaling in C2C12 Myotubes

 

The dosages of reagents were 0.5 mM for leucine, 0.5 mM for alanine, 0.5 mM for valine...

Leucine significantly increased mitochondria content in C2C12s compared to alanine and valine.

 

Edited by Turnbuckle, 12 March 2018 - 07:50 PM.

[BigLabRat]

No, I suspect he means "Leucine." The three branched-chain amino acids (BCAAs)--leucine, isoleucine, and valine--have powerful effects on muscle building, and are being found to have other anabolic effects as well.

 

5 grams of leucine isn't tiny by any means. It takes 10 eggs to get you 5 grams of leucine. Or two-thirds of a pound of beef. A fairly protein-rich meal like a three-egg cheese omelette gives you around 2.5 g of leucine.

 

So, consuming 5 g of leucine is like eating two extra three-egg cheese omelettes a day. Or an extra 1 1/3 pounds of beef. It's a big boost in intake.

 

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

 

Milk-based products tend to be high in the BCAAs, which is why so many bodybuilders consume whey protein. Whey powder is about 10% leucine by weight.

 

Some whey products aimed at the bodybuilding crowd add extra leucine. But Jeff Volek's work (Applied Physiology, Nutrition and Metabolism. 2009 Apr; Vol. 34, No. 2, 151-61.) shows that the stimulating effects of leucine top out pretty quickly. So an additional 5 g of leucine is bound to saturate the system of someone eating normally.

 

I'd view 5 g of supplemental leucine as an insurance policy. There's no contraindications. But there's quite likely no benefit in going higher, either...

 

[BigLabRat]

Hey, Turnbuckle,

 

Thanks for laying out the most recent protocols in detail.

 

I have one question: What's your current thinking on how these protocols interact with fasting? I do moderate (3-7 days) fasts a few times a year. Would there be benefits to combining fasting with the fission protocol?

 

Since fasting seems to promote various forms of autophagy, it seems sensible at first. But Walter Longo's work on fasting and chemotherapy suggests that fasting causes 'healthy' cells to 'hunker down' into a sort of hibernation, while cancer cells continue on about their business. This leaves the cancer cells vulnerable to chemo, but protects the healthy cells.

 

I'm trying to sort out how this works in the case of mitophagy. Would fasting be synergetic with the fission protocol--or would fasting tend to make the cells less active, and thereby neutralize the protocol?

 

Or are we off in the realm where there just isn't enough evidence?

[Turnbuckle]

Hey, Turnbuckle,

 

Thanks for laying out the most recent protocols in detail.

 

I have one question: What's your current thinking on how these protocols interact with fasting? I do moderate (3-7 days) fasts a few times a year. Would there be benefits to combining fasting with the fission protocol?

 

Since fasting seems to promote various forms of autophagy, it seems sensible at first. But Walter Longo's work on fasting and chemotherapy suggests that fasting causes 'healthy' cells to 'hunker down' into a sort of hibernation, while cancer cells continue on about their business. This leaves the cancer cells vulnerable to chemo, but protects the healthy cells.

 

I'm trying to sort out how this works in the case of mitophagy. Would fasting be synergetic with the fission protocol--or would fasting tend to make the cells less active, and thereby neutralize the protocol?

 

Or are we off in the realm where there just isn't enough evidence?

 

 

Given the following paper, I'd say fission/mitophagy works best without fasting and with non-endurance exercise. I usually do the fission protocol early in the morning on an empty stomach, working out with weight machines (mostly) 2-3 hours later, and don't eat till I get back. I wouldn't call this fasting, however, and certainly not endurance.

 

 

CONCLUSION:

The present study provides the very first evidence that mitophagy is not activated during and early after high-intensity endurance exercise in human, whatever the nutritional state, despite a selective activation of fission in the fed state. However, when nutrient availability is optimal, muscle cells seem capable of preparing mitochondria for lysosomal degradation. Thus, we may not exclude an activation of mitophagy at a later stage after exercise.

https://www.ncbi.nlm...pubmed/28272266

 

 

 

Edited by Turnbuckle, 12 March 2018 - 08:15 PM.

[BigLabRat]

Very interesting, thanks.

 

The plot thickens.

[Nate-2004]

Now I'm even more confused, fission really is not easy to achieve I don't think. Too many contradicting complications. Just about everything you eat has at least a good 2g of stearic acid per serving. A LOT of foods have leucine in it. Yet simply fasting to avoid these things doesn't enable fission all that well so that's a waste of time.  You can't really win at this I don't think. Not really. I pretty much do exactly what Turnbuckle does except maybe no direct consumption of stearic acid or AMPK activators, other than berberine.

Edited by Nate-2004, 12 March 2018 - 10:04 PM.

[BigLabRat]

Yes, leucine is found in virtually all proteins. That said, you don't often encounter 5 grams of leucine at one go without taking supplements.

 

The issue of fasting isn't a matter of eliminating stearic acid or leucine. Fasting is a specialized metabolic state, especially after 72 hours.

 

Fasting clearly accelerates general autophagy. Whether it accelerates or slows mitophagy in particular doesn't seem clear. Fasting also 'reboots' the immune system, replacing the existing cadre of cells with new cells.

 

We don't yet understand all of what fasting does at a cellular level, but it's clearly quite powerful. So I'm trying to understand how fasting might or might not interact with these protocols.

[Nate-2004]

I understand the whole idea isn't to avoid stearic acid or leucine but honestly it's the only way I could think of aside from eating nothing but spinach that day, which even still contains 1mg of stearic acid.

[Turnbuckle]

I understand the whole idea isn't to avoid stearic acid or leucine but honestly it's the only way I could think of aside from eating nothing but spinach that day, which even still contains 1mg of stearic acid.

 

Stearic acid isn't as strong as you think, and luciene isn't going to be a problem. If you are having trouble getting enough fission, add in 100mg apigenin. 

 

Tributyltin induces mitochondrial fission through NAD-IDH dependent mitofusin degradation in human embryonic carcinoma cells

Staining with MitoTracker revealed that nanomolar TBT levels induced mitochondrial fragmentation. TBT also degraded the mitochondrial fusion proteins, mitofusins 1 and 2. Interestingly, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT.

http://pubs.rsc.org/...3e#!divAbstract

 

Edited by Turnbuckle, 13 March 2018 - 01:32 PM.

[matrix83]

May be of some interest:

 

https://www.frontier....00086/abstract

 

Ketogenic diet improves brain ischemic tolerance and inhibits NLRP3 inflammasome activation by preventing Drp1-mediated mitochondrial fission and endoplasmic reticulum stress

[BigLabRat]

Interesting abstract. But I'd need to see the whole paper (not yet published) to figure out what they're talking about!

[zorba990]

I understand the whole idea isn't to avoid stearic acid or leucine but honestly it's the only way I could think of aside from eating nothing but spinach that day, which even still contains 1mg of stearic acid.

Stearic acid isn't as strong as you think, and luciene isn't going to be a problem. If you are having trouble getting enough fission, add in 100mg apigenin.

Tributyltin induces mitochondrial fission through NAD-IDH dependent mitofusin degradation in human embryonic carcinoma cells
Staining with MitoTracker revealed that nanomolar TBT levels induced mitochondrial fragmentation. TBT also degraded the mitochondrial fusion proteins, mitofusins 1 and 2. Interestingly, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT.
http://pubs.rsc.org/...3e#!divAbstract

Just fyi 100mg apigenin with 3g Niacinamide + 5g ribose was too much for me. I got really ill both times I tried it and was wiped out for almost 2 days. I don't think the follow on fusion resulted in a net gain for me...so I'm going to go more slowly since I have to work :-) I took the same without the apigenin and just got the usual am tiredness that faded by early afternoon.

[Empiricus]

 

 

This is no typo, and I suggest that leucine be used alone as including other amino acids can dilute or eliminate the desired effect. For the same reason stearic acid should be used alone and not in some mix with FAs, as some FAs can cause mito uncoupling.

 

 

Hmmm.  The fact I've never taken stearic acid on its own might explain why I seldom experience much bang from the protocol.  Anyway, I've ordered some.  

Edited by Empiricus, 14 March 2018 - 03:12 AM.