2284 replies to this topic

[Turnbuckle]

 

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.

 

 

3g nicotinamide + an excess of ribose is a lot, equivalent to 6g NR. And apigenin has a long half-life--reported as 92 hours. So I suggest starting with one or two grams nicotinamide + a like amount of ribose (an excess doesn't seen to be necessary), and adding in the apigenin only if you don't feel you're getting much fission (and apigenin only on the first day of fission). The N+R protocol can knock down your BP, especially if you have a lot of damaged mitochondria. Early on, my systolic pressure (180+ without drugs) was knocked down to below a hundred and remained at 120 or below for a week. I was having a hard time staying awake. Now a year later this effect is much smaller, but my BP is almost always lower with fission and higher with fusion--around ten points or so either way.

[Nate-2004]

 

 

 

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.  

 

 

Maybe, I find it hard to avoid FAs in general, most people's diets have a lot of FAs, whether the good or bad kind. Maybe taking the stearic on an empty stomach might help, might not.

[Nate-2004]

I ordered some straight apigenin from swanson, which I've done before a year or two ago. I don't know that I trust Swanson but they're the only brand out there that sells it. I would love to know exactly how much apigenin comes in your average 12 oz glass of chamomile tea with one bag and/or two because I sleep great on that and it's guaranteed to have it in there. I have a ton of that stuff. 

 

Yeah to the previous poster on being wiped out, I think you're taking too much ribose. I don't know how old you are but try to make it an even ratio. I tend to make it a 5:4 ratio NAM:R. Maybe that's my issue though.

Edited by Nate-2004, 14 March 2018 - 12:54 PM.

[Turnbuckle]

 

Yeah to the previous poster on being wiped out, I think you're taking too much ribose. I don't know how old you are but try to make it an even ratio. I tend to make it a 5:4 ratio NAM:R. Maybe that's my issue though.

 

 

I don't think ribose is the problem. It's more likely the threshold effect that some individuals will have due to high levels of defective mitochondria that they are unaware of. One might have 80% dysfunctional mtDNA loops and the mitochondria still function. Mitochondria typically contain several loops that cover for each other, and thus cells still have enough ATP to operate normally. But if you fragment the mitochondria to minimal size with one loop each, the threshold is lowered and all the bad mtDNA loops are exposed as nonfunctional. ATP production drops off a cliff and cells may be overwhelmed with too many dysfunctional mitochondria to process, as lysosomes are limited. Thus best to start with a low level N+R and build it up. And if you get wiped out on the first day, don't continue with fission for the next day. Cut the cycle short and drop the N+R level the next time. It may take many cycles of fission/mitophagy and fusion/biogenesis to make a substantial dent on the dysfunctional population.

Edited by Turnbuckle, 14 March 2018 - 02:18 PM.

[aribadabar]

Fission + mitophagy (2-3 days)

Nicotinamide (NAD+) — 2g

Ribose (NAD+) — 2g

AMPK activators (Life Extension) — 1g 

Fisetin (Sirt1 activator) — 100mg

 

Can I use apigenin instead of Fisetin in the Fission phase? I read that they have similar MOA and I have some.

And if yes, is substitution 1:1 or different (more/less)?

[Nate-2004]

I'm going to try adding an extra day in between fusion and fission and start ramping up a little with NR and apigenin. Then on the next day go hard with it, NAM+R, NR and Apigenin, along with the anaerobic exercise routine which for me is deadlifts, push ups, leg curls and pull ups mainly.

 

Still not sure why the AMPK activators. It seems there's been some back and forth about that on this thread early on. This thread has become a bit unruly. I really wish you could search individual threads for key words or just load all pages into one and do a browser find.

[BieraK]

 

Fission + mitophagy (2-3 days)

Nicotinamide (NAD+) — 2g

Ribose (NAD+) — 2g

AMPK activators (Life Extension) — 1g 

Fisetin (Sirt1 activator) — 100mg

 

Can I use apigenin instead of Fisetin in the Fission phase? I read that they have similar MOA and I have some.

And if yes, is substitution 1:1 or different (more/less)?

 

 

I have the same question about using apigenin instead of fisetin.

I wonder also if the stack could be done with less supplements, for example, just using AMPK activators and N+R for fission and using just Stearic Acid and PQQ and removing Hydroxytyrosol and Leucine for fusion. Tyrosol is a Sirt1 activator also, like Fisetin, so in both Fission and Fusion state Sirt1 is activated?.
However some of these pathways shares routes with each other, PQQ activates Sirt1, and Sirt1 activation with AMPK activates PGC-1a.

The dose from AMPK Life Extension gynostemma is very high, I have 98% Gypenosides Jiaogulan extract and 200 mg is enough to produce fatigue.

[whileitravel]

While watching a Rhonda Patrick video of her interviewing Dr. Eric Verdin, he indicated that they were able to show ear nerve regeniration in mice, where they could hear once again. 

 

I'm getting what can only be described as a tingling sensation in my right ear the last couple of weeks while trying this protocol. Has anyone else experienced this?

Edited by whileitravel, 15 March 2018 - 02:23 AM.

[Turnbuckle]

 

 

 

I have the same question about using apigenin instead of fisetin.
 

 

How are apigenin and fisetin interchangeable? I included fisetin as a sirtuin activator as sirtuins activate the PINK/PARKIN pathway that leads to mitochondrial fission and mitophagy. If you want to replace it, you could possibly use resveratrol instead. I don't use resveratrol as it causes me joint pain (and almost crippled me some years ago). As for apigenin, I originally included it as a strong fission promoter, but then took it out because of its long half life (92 hours). Those who aren't getting enough fission just from N+R and fisetin might want to use it, in particular on the first day of fission when fusion is still several days off. 

 

Note: The protocols I post on this thread I've tested on myself, but no one else. Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

Active autophagy coupled with rapid mitochondrial fusion and fission constitutes an important mitochondrial quality control mechanism and is critical to cellular health. In our previous studies, we found that exposure of cells to nicotinamide causes a decrease in mitochondrial content and an increase in mitochondrial membrane potential (MMP) by activating autophagy and inducing mitochondrial fragmentation. Here, we present evidence to show that the effect of nicotinamide is mediated through an increase of the [NAD+]/[NADH] ratio and the activation of SIRT1, an NAD+-dependent deacetylase that plays a role in autophagy flux. The [NAD+]/[NADH] ratio was inversely correlated with the mitochondrial content, and an increase in the ratio by the mobilization of the malate-aspartate shuttle resulted in autophagy activation and mitochondrial transformation from lengthy filaments to short dots. Furthermore, treatment of cells with SIRT1 activators, fisetin or SRT1720, induced similar changes in the mitochondrial content. Importantly, the activators induced mitochondrial fragmentation only when SIRT1 expression was intact. Meanwhile, MMP did not increase when the cells were treated with the activators, suggesting that the change in MMP is not induced by the mitochondrial turnover per se and that elevation of the [NAD+]/[NADH] ratio may activate additional mechanisms that cause MMP augmentation. Together, our results indicate that a metabolic state resulting in an elevated [NAD+]/[NADH] ratio can modulate mitochondrial quantity and quality via pathways that may include SIRT1-mediated mitochondrial autophagy.

https://www.ncbi.nlm...les/PMC3365962/

 

Edited by Turnbuckle, 15 March 2018 - 01:37 PM.

[aribadabar]

Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

I started the fission phase yesterday and it is unmistakable - I start feeling fairly anxious a couple of hours after 2g N+2g R+100mg fisetin+4x250mg  (1g) caps of this gynostemma extract , both today and yesterday. It's tolerable but certainly unpleasant and lasts for several hours.

Is that showing that it is working as intended or a sign of "overdosing" and if the latter, should I cut the dose of all of just skip some of the supplementary items (fisetin, gynostemma) completely?

Edited by aribadabar, 15 March 2018 - 08:30 PM.

[whileitravel]

 

Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

I started the fission phase yesterday and it is unmistakable - I start feeling fairly anxious a couple of hours after 2g N+2g R+100mg fisetin+4x250mg  (1g) caps of this gynostemma extract , both today and yesterday. It's tolerable but certainly unpleasant and lasts for several hours.

Is that showing that it is working as intended or a sign of "overdosing" and if the latter, should I cut the dose of all of just skip some of the supplementary items (fisetin, gynostemma) completely?

 

I'm sorry to hear that you've had a not so pleasent reaction. Apart from the tingling in my right ear that I posted here yesterday, I've experienced no positive or negative effects. I've been using the protocol apart from the stearic acid for a period of greater than 2 months.

 

I previously used NR and I think this here is the best bang for the buck.

 

FWIW I'm 51 and in excellent health.

 

 

 

[whileitravel]

 

Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

I started the fission phase yesterday and it is unmistakable - I start feeling fairly anxious a couple of hours after 2g N+2g R+100mg fisetin+4x250mg  (1g) caps of this gynostemma extract , both today and yesterday. It's tolerable but certainly unpleasant and lasts for several hours.

Is that showing that it is working as intended or a sign of "overdosing" and if the latter, should I cut the dose of all of just skip some of the supplementary items (fisetin, gynostemma) completely?

 

I'm sorry to hear that you've had a not so pleasent reaction. Apart from the tingling in my right ear that I posted here yesterday, I've experienced no positive or negative effects. I've been using the protocol apart from the stearic acid for a period of greater than 2 months.

 

I previously used NR and I think this here is the best bang for the buck.

 

FWIW I'm 51 and in (knock on wood) excellent health.

 

 

 

[Turnbuckle]

Apart from the tingling in my right ear that I posted here yesterday, I've experienced no positive or negative effects. I've been using the protocol apart from the stearic acid for a period of greater than 2 months.

 

I previously used NR and I think this here is the best bang for the buck.

 

 

 

 

I don't understand this. You've had no positive or negative effects, and you think it is the best bang for the buck?

Edited by Turnbuckle, 15 March 2018 - 11:04 PM.

[whileitravel]

Hi Turnbuckle. It may sound absurd I'm sure, but I'm assuming since I am "young" and healty the effects of NR are minimal. I'm thinking the true effects will be more long term down the road.

 

I want to believe that taking NR at my age is beneficial and once I saw this protocol I stopped buying Niagen and began trying this protocol.

 

Hope that makes sense.

[William Sterog]

I don't remember whether TUDCA has been mentioned in this thread or not.

 

Tauroursodeoxycholic Acid Enhances Mitochondrial Biogenesis, Neural Stem Cell Pool, and Early Neurogenesis in Adult Rats.

https://www.ncbi.nlm...pubmed/28534273

 

More promising information on TUDCA: 

 

Researchers determine cause of difficult-to-control mitochondrial diseases

https://www.news-med...l-diseases.aspx

 

Edited by William Sterog, 16 March 2018 - 04:12 PM.

[Turnbuckle]

 

I don't remember whether TUDCA has been mentioned in this thread or not.

 

Tauroursodeoxycholic Acid Enhances Mitochondrial Biogenesis, Neural Stem Cell Pool, and Early Neurogenesis in Adult Rats.

https://www.ncbi.nlm...pubmed/28534273

 

More promising information on TUDCA: 

 

Researchers determine cause of difficult-to-control mitochondrial diseases

https://www.news-med...l-diseases.aspx

 

 

 

It also attenuates amyloid precursor protein processing and amyloid-β deposition in mice--

 

Consequently, a significant decrease in Aβ(1-40) and Aβ(1-42) levels was observed in both hippocampus and frontal cortex of TUDCA-treated APP/PS1 mice, suggesting that chronic feeding of TUDCA interferes with Aβ production, possibly through the regulation of lipid-metabolism mediators associated with APP processing. These results highlight TUDCA as a potential therapeutic strategy for the prevention and treatment of AD.

https://www.ncbi.nlm...pubmed/22438081

 

[BigLabRat]

TUDCA is a very promising supplement in general. Here's a general review:

 

https://examine.com/...oxycholic-acid/

 

That said, while it apparently increases mitochondrial biogenesis, it also triggers cellular apoptosis, and has a number of other effects.

 

Curiously, it is hepatoprotective if taken after drinking, but increases liver damage if ingested before drinking. All in all, it's a curious and potent molecule, and it's clear there's still a lot to learn about its (sometimes conflicting) actions.

 

[aribadabar]

 

Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

I started the fission phase yesterday and it is unmistakable - I start feeling fairly anxious a couple of hours after 2g N+2g R+100mg fisetin+4x250mg  (1g) caps of this gynostemma extract , both today and yesterday. It's tolerable but certainly unpleasant and lasts for several hours.

Is that showing that it is working as intended or a sign of "overdosing" and if the latter, should I cut the dose of all of just skip some of the supplementary items (fisetin, gynostemma) completely?

 

 

On day 3 of fission, I decided to change the protocol and skipped taking gynostemma extract while keeping the rest unchanged.

FWIW today the anxious feeling is still palpable but significantly reduced.

[BieraK]

 

 

 

 

I have the same question about using apigenin instead of fisetin.
 

 

How are apigenin and fisetin interchangeable? I included fisetin as a sirtuin activator as sirtuins activate the PINK/PARKIN pathway that leads to mitochondrial fission and mitophagy. If you want to replace it, you could possibly use resveratrol instead. I don't use resveratrol as it causes me joint pain (and almost crippled me some years ago). As for apigenin, I originally included it as a strong fission promoter, but then took it out because of its long half life (92 hours). Those who aren't getting enough fission just from N+R and fisetin might want to use it, in particular on the first day of fission when fusion is still several days off. 

 

Note: The protocols I post on this thread I've tested on myself, but no one else. Results will vary between users, so feel free to experiment with different ingredients and post your results.

 

Active autophagy coupled with rapid mitochondrial fusion and fission constitutes an important mitochondrial quality control mechanism and is critical to cellular health. In our previous studies, we found that exposure of cells to nicotinamide causes a decrease in mitochondrial content and an increase in mitochondrial membrane potential (MMP) by activating autophagy and inducing mitochondrial fragmentation. Here, we present evidence to show that the effect of nicotinamide is mediated through an increase of the [NAD+]/[NADH] ratio and the activation of SIRT1, an NAD+-dependent deacetylase that plays a role in autophagy flux. The [NAD+]/[NADH] ratio was inversely correlated with the mitochondrial content, and an increase in the ratio by the mobilization of the malate-aspartate shuttle resulted in autophagy activation and mitochondrial transformation from lengthy filaments to short dots. Furthermore, treatment of cells with SIRT1 activators, fisetin or SRT1720, induced similar changes in the mitochondrial content. Importantly, the activators induced mitochondrial fragmentation only when SIRT1 expression was intact. Meanwhile, MMP did not increase when the cells were treated with the activators, suggesting that the change in MMP is not induced by the mitochondrial turnover per se and that elevation of the [NAD+]/[NADH] ratio may activate additional mechanisms that cause MMP augmentation. Together, our results indicate that a metabolic state resulting in an elevated [NAD+]/[NADH] ratio can modulate mitochondrial quantity and quality via pathways that may include SIRT1-mediated mitochondrial autophagy.

https://www.ncbi.nlm...les/PMC3365962/

 

 

 

because in a previous comment you showed how apigenin induces mitochondrial fission.

According to this quote, Fisetin activating Sirt1 induces autophagy. As I have learned about this topic (that is pretty little) the mitochondrial fission alone does not ensure mitophagy, the activation of the autophagic pathways and mitochondrial fission is necessary for the mitophagy to occur, so thats the reason behind AMPK from gynostemma and Sirt1 from Fisetin. I was trying to know if apigenin activates Sirt1 but nothing happened, the closest research is this: https://www.ncbi.nlm...pubmed/28143892

So if I'm understaing this correctly, this protocol is like taking NR+Pterostilbene (Elysium Basis) in a massive dose after a 12 hour overnight fasting (for the reduced glucose and glycogen leading to AMPK) but in a cheaper way.
 

[BieraK]

Hi Turnbuckle. It may sound absurd I'm sure, but I'm assuming since I am "young" and healty the effects of NR are minimal. I'm thinking the true effects will be more long term down the road.

 

I want to believe that taking NR at my age is beneficial and once I saw this protocol I stopped buying Niagen and began trying this protocol.

 

Hope that makes sense.

 

I agree with the opinion of Luigi Fontana, is best to change the idea of treating illness for a medicine based on a preventive perspective. Fontana is an expert on calorie restriction and Judging by what he looks like for his 50 years, he seems to be on a calorie restriction diet.
So, it is better to take NR for prevention and for slowing age, I do not agree with the idea that, if there is nothing to repair there is nothing to intervene, thats works only for people with a good lifestyle and the good genes.

 

[ceridwen]

Fisetin also has a beneficial effect on AD because it reduces methylglyoxal one of the drivers of diabetes.

[Nate-2004]

There is simply not enough research and thus evidence on whether pterostilbene does anything similar to resveratrol. On top of that resveratrol has low bioavailability due to certain enzymes.

 

Theoretically, apigenin and resveratrol both compete for these enzymes, which means that they could be very synergistic together in terms of improving the length of time in circulation. Is it that resveratrol improves apigenin bioavailability, or is it an improvement with both?

 

https://www.ncbi.nlm...les/PMC4663613/

https://www.scienced...021949816300965

 

There's the suggestion that it does in this Google book scan, not sure what this is though:

 

https://books.google...ability&f=false

 

[zorba990]

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.

3g nicotinamide + an excess of ribose is a lot, equivalent to 6g NR. And apigenin has a long half-life--reported as 92 hours. So I suggest starting with one or two grams nicotinamide + a like amount of ribose (an excess doesn't seen to be necessary), and adding in the apigenin only if you don't feel you're getting much fission (and apigenin only on the first day of fission). The N+R protocol can knock down your BP, especially if you have a lot of damaged mitochondria. Early on, my systolic pressure (180+ without drugs) was knocked down to below a hundred and remained at 120 or below for a week. I was having a hard time staying awake. Now a year later this effect is much smaller, but my BP is almost always lower with fission and higher with fusion--around ten points or so either way.

Thanks. I beleive apigenin has a 12 hr half life though. From pubchem:
https://pubchem.ncbi...mpound/apigenin

“Apigenin appears to be absorbable by humans after intake of parsley (Petroselinum crispum). In a randomized crossover study with two one-week intervention periods in succession, fourteen volunteers consumed a diet that included 20 g parsley. The urinary excretion of apigenin was significantly higher (P < 0.05) during the intervention with parsley (20.7 to 5727.3 g/24 hr) than during the basic diet (0 to 1571.7 g/24 hr). The half-life for apigenin was calculated to be on the order of 12 hr“

And your other post:
http://www.longecity...life-918-hours/

[Turnbuckle]

Yep, zorba. I seem to have forgotten that post. Thanks for reminding me.

Edited by Turnbuckle, 18 March 2018 - 08:25 PM.

[Nate-2004]

92 vs 12 is a big difference in half-life, I wish we had a more definitive answer on that.

 

Also back to my resveratrol with apigenin post above, don't do it at night, it'll keep you awake. While apigenin helps me sleep, whether pills or chamomile, the resveratrol will keep you awake I think. I don't know if this was some one off issue I had last night or what. I'm also not sure what will happen if I take apigenin during the day, it's usually something I take before bed.

[William Sterog]

"We here report that piracetam at therapeutically relevant concentrations improves neuritogenesis in the human cell line SH-SY5Y over conditions mirroring the whole spectrum of age-associated cognitive decline. These effects go parallel with improvement of impaired mitochondrial dynamics shifting back fission and fusion balance to the energetically more favorable fusion site. Impaired fission and fusion balance can also be induced by a reduction of the mitochondrial permeability transition pore (mPTP) function as atractyloside which indicates the mPTP has similar effects on mitochondrial dynamics. These changes are also reduced by piracetam."

https://www.ncbi.nlm...am mitochondria

[aribadabar]

"We here report that piracetam at therapeutically relevant concentrations improves neuritogenesis in the human cell line SH-SY5Y over conditions mirroring the whole spectrum of age-associated cognitive decline. These effects go parallel with improvement of impaired mitochondrial dynamics shifting back fission and fusion balance to the energetically more favorable fusion site. Impaired fission and fusion balance can also be induced by a reduction of the mitochondrial permeability transition pore (mPTP) function as atractyloside which indicates the mPTP has similar effects on mitochondrial dynamics. These changes are also reduced by piracetam."

https://www.ncbi.nlm...am mitochondria

 

1 mM concentration - how much piracetam one needs to ingest to reach this level in humans?

[Krell]

I am on my second week of fission/fusion protocols, as in

http://www.longecity...-28#entry843333

 

But I am running into a problem with my heart rate.

 

When I take 1.0g D-Ribose + 1.0g nicotinamide or more, my heart rate during aerobic exercise

becomes irregular according to my heart rate monitor.  Although I feel normal

I seem to be getting some extra beats. I am doing HIIT on a Matrix stair-stepper that

has a built-in HR monitor that senses HR from your hands during exercise.  Never had

this problem before in several years of using this equipment.  There is no problem at

lower levels of exercise, but the problem gets worse at higher stress levels.  

 

Anyone else had this problem? 

 

Do you need to start dosage at low levels and build up over days/weeks?

 

(73yro 180lb 6ft fit) 

 

[Turnbuckle]

I am on my second week of fission/fusion protocols, as in

http://www.longecity...-28#entry843333

 

But I am running into a problem with my heart rate.

 

When I take 1.0g D-Ribose + 1.0g nicotinamide or more, my heart rate during aerobic exercise

becomes irregular according to my heart rate monitor.  Although I feel normal

I seem to be getting some extra beats. I am doing HIIT on a Matrix stair-stepper that

has a built-in HR monitor that senses HR from your hands during exercise.  Never had

this problem before in several years of using this equipment.  There is no problem at

lower levels of exercise, but the problem gets worse at higher stress levels.  

 

Anyone else had this problem? 

 

Do you need to start dosage at low levels and build up over days/weeks?

 

(73yro 180lb 6ft fit) 

 

I haven't had this problem, but I've stuck to fission for weights and fusion for aerobics.

[QuestforLife]

I am on my second week of fission/fusion protocols, as in

http://www.longecity...-28#entry843333

 

But I am running into a problem with my heart rate.

Makes sense. The heart has more mitochondria per cell than anywhere else, and you're making it work hard whilst it has a significantly reduced number of mitochondria.

 

I'd do the protocol a few more times before you try HIIT again. I do it okay, but I've done Turnbuckle's protocol 10+ times, and I'm only 39 so probably have less bad mitos to clear.