Alternative methods to extend telomeres

As far as I understood, Questforlife, I think the idea is to use cryo-electron microscopy to identify the structure of HTERT activators and repressors and later create bio-identicals to use respectively as longevity and anti-cancer medication. Scan for activators in cancer cells with telomerase activity. Scan for repressor proteins in regular somatic cells.

Yes, that would be useful - providing the various sites can be identified, any resulting proteins or splicing factors could be examined and identified and either blocked or replicated with small molecules to get the required outcome: either telomerase inhibition or activation. I don't know how plausible achieving the first step is however: the identification of the relevant sites on the DNA, or how microscopy would help with that. It might be easier to examine splicing factors first in various cellular scenarios. Splicing factors are the switches that alter the form of a given protein that is produced, and its resultant functionality. It's very complicated to unpick all the splicing factors however.

One alternative might be to make a synthetic molecule with the shape of telomerase and then find a way to get it into every cell. But then why not just do gene therapy instead?

These difficulties are why I am pursuing 'Alternative methods to extend telomeres', with the current emphasis on progenitor cell generation (which have less restricted telomerase) along with telomerase activation. This has the added bonus of also reducing epigenetic age, which may also be beneficial.

Edited by QuestforLife, 07 January 2019 - 10:20 AM.

Thank you for the opportunity to spell out my protocol as it stands. In November I took 10mg of atorvastatin and 25mg of losartan along with 100mg of Q10, all once per day in the morning, for 1-month.

Since then I have taken almost no supplements with the intention of repeating the protocol in February or March. In the original Janic etc al. work posted upthread the interval was 6 months (or in with the diabetics study 3 months). I believe this is based on the time for the progenitor cells to fully differentiate back into normal cells. I am considering a different interval however, based on 2 weeks on and 6 weeks off. This is based on the following paper where they used ROCK and mTOR inhibitors to reprogram glioblastoma cells (in 3-7days), and differentiate then into normal neurons (in 2-3 weeks). I've doubled their timescales because I know detectable ROCK inhibition takes about 2 weeks (in Vivo).

https://jeccr.biomed...3046-018-0857-5

I am also considering using rapamycin during the protocol as well to enhance the reprogramming efficiency. If I go with a 2 week protocol rather than 1-month doses will also be higher than what I used previously. I will post these once I finalise them.

I believe the benefits are largely lost once the progenitor cells differentiate because the somatic cells go back to being the age they were before de-differentiation, with the same length telomeres. For this reason I will include cyclastragneol in the protocol to try and lengthen telomeres whilst the progenitor cells are active.
https://www.ncbi.nlm...ubmed/24774536/

As you can tell the protocol is still in flux, but it is very promising.

One other note, I believe that some of the benefits of the protocol overlap with those of a ketogenic diet, because Statins cause upregulation of fatty acid oxidation. Since late November I've been ketogenic. This is not strictly necessary (Janic etc al. didn't do this), but I believe it may help. Just something to consider.

The end point for my n=1 study is another (my third) Zymo epigenetic age test next summer.

High Desert Wizard brough to my attention the effects on bee keeping and honey on telomeres, see this paper where beekeepers had significantly longer telomeres than the age matched (actually younger) control group:

https://www.ncbi.nlm...rticle_9797.pdf

Interestingly royalactin (the most common protein in royal jelly and the one responsible for differentiating queen bees from drones ) has been shown to maintain stemness, at least in vitro:

https://www.nature.c...467-018-06256-4

Very rough calculation, this would take ATLEAST 15g of royal jelly a day to replicate the concentration in vivo, so I'm not certain I'm willing to take that risk to add it to my protocol just yet (only seen a human study where they took up to 3g a day).

High Desert Wizard brough to my attention the effects on bee keeping and honey on telomeres, see this paper where beekeepers had significantly longer telomeres than the age matched (actually younger) control group:

 

https://www.ncbi.nlm...rticle_9797.pdf

 

Interestingly royalactin (the most common protein in royal jelly and the one responsible for differentiating queen bees from drones ) has been shown to maintain stemness, at least in vitro:

 

https://www.nature.c...467-018-06256-4

 

Very rough calculation, this would take ATLEAST 15g of royal jelly a day to replicate the concentration in vivo, so I'm not certain I'm willing to take that risk to add it to my protocol just yet (only seen a human study where they took up to 3g a day).

i don't the time to read the articles atm but is it possible that its bee venom causing the telomere lengthening effect? Steve Ludwin claims he had the telomere age of 28 at 46 years old. He injects snaked venom very day for the past 30 years/

i don't the time to read the articles atm but is it possible that its bee venom causing the telomere lengthening effect? Steve Ludwin claims he had the telomere age of 28 at 46 years old. He injects snaked venom very day for the past 30 years/

 

I beleive that the answer to your question is no, because they found telomere length to be correlated with the number of bee products consumed per day - so having honey multiple times is much better than once. There was also no link with the time spent being a bee keeper, so likely it is not to do with exposure to bee toxin. That's not to say that toxins or indeed snake toxin has no influence on telomeres, there is just no evidence for it in this study. 

Quest, where do you purchase your atorvastatin and losartan?

aipctshop.

I'd recommend this protocol to anyone concerned about atherosclerosis and approaching middle age or older, or younger but with diabetes, as that is where the results have been proven by Janic et al.  I suspect the benefits may well go further than the vascular system, but that has not yet been demonstrated.

I have started a new protocol based on my continuing research into ROCK inhibition/progenitor cells and telomeres/cellular senescence.

The protocol is highly experimental, and until I have some results to report I do not advise anyone else try this. I believe it is safe as I have separately used most of the treatments of the protocol before without harm. This is the first time I’ve used them as a combinatorial treatment, however.

It is a repeating 54 day (~2 month) cycle with overlapping Phases.

1. Phase 1 is Senolytics, currently 5 days of azithromycin with a loading dose of 2g and a maintenance dose of 1g on subsequent days.

2. Phase 2 is progenitor cell generation, currently 14 days of 10mg atorvastatin, 25mg losartan, 30mg pioglitazone daily. Phase 2 overlaps with the last 3 days of Phase 1, to aid with the expansion of progenitor cell numbers.

3. Phase 3 is 28 days of telomere stimulation, currently 25mg of cycloastragenol, 100mg bioavailable Q10, 1.5g royal jelly (equivalent). Phase 3 overlaps with the last 11 days of Phase 2 in order to make the most of the more active HTERT site on the new progenitor cells, before they have all differentiated (expected in 2-3 weeks from the end of Phase 2).

4. Phase 4 is a 21 day wash-out/differentiation phase.

Then the protocol begins again with Phase 1, possibly with a different senolytic. I will do it up to 6 times in 2019. Endpoints will be measured by MyDNAge epigenetic age testing (have baseline from 2017 and 2018), Lifelength telomere testing (have baseline from 2018), exercise tolerance, physical appearance, and changes in any standard blood markers (have baseline from 2015 and 2018).

I have references and reasoning for all Phases 1-4, and welcome questions. Refinements are possible.

Background lifestyle is sedentary office job, daily mediation, ketogenic or low-carb diet, weight lifting and occasional high intensity interval training. All of these have been in place unchanged since Nov 2018.

Edited by QuestforLife, 22 January 2019 - 03:30 PM.

As strange as it seems, there actually is support for alternative lengthening of telomeres with Royal Jelly through activating the Akt signaling pathway.

1. “Royal Jelly Prevents the Progression of Sarcopenia in Aged Mice In Vivo and In Vitro” (2013) https://drive.google...5krH_erke/view 
2. “IL-2 Increases Human Telomerase Reverse Transcriptase Activity Transcriptionally and Posttranslationally through Phosphatidylinositol 3′-Kinase/Akt, Heat Shock Protein 90, and Mammalian Target of Rapamycin in Transformed NK Cells” (2017) http://www.jimmunol....ent/174/9/5261 

aipctshop.

I'd recommend this protocol to anyone concerned about atherosclerosis and approaching middle age or older, or younger but with diabetes, as that is where the results have been proven by Janic et al. I suspect the benefits may well go further than the vascular system, but that has not yet been demonstrated.

I have started a new protocol based on my continuing research into ROCK inhibition/progenitor cells and telomeres/cellular senescence.

 

It is a repeating 54 day (~2 month) cycle with overlapping Phases.

 

1. Phase 1 is Senolytics, currently 5 days of azithromycin with a loading dose of 2g and a maintenance dose of 1g on subsequent days.

 

2. Phase 2 is progenitor cell generation, currently 14 days of 10mg atorvastatin, 25mg losartan, 30mg pioglitazone daily. Phase 2 overlaps with the last 3 days of Phase 1, to aid with the expansion of progenitor cell numbers.

 

Observations from Phase I (complete): previously when I tried out azithromycin with a loading dose of 1g, I felt very tired for an hour or two on the first day, but otherwise noticed nothing (actually I felt remarkably good during the 10 days I took it for). This time with a loading dose of 2g and a maintenance dose of 1g I felt nothing on day 1, but on day 2 I felt anxious for some undefinable reason. Later I realized I was short of breath, which is unusual for me. The feeling had passed by the end of the day. I felt normal for the rest of the 5 days.

Observations from Phase 2 (half way through): once again my skin looks great.

Possible explanation is the extracellular vesicles released by progenitor cells?  

https://www.ncbi.nlm...pubmed/30598033

Experimental data indicate that human endothelial progenitor cells EVs from umbilical cord stem cells accelerate cutaneous wound healing in diabetic rats by promoting endothelial function [79]. Moreover, EVs enhanced the proliferation, migration, and tube formation of vascular endothelial cells in vitro and even stimulated the production of FGF-1, VEGFA, VEGFR-2, Angiopoietin 1 (ANG-1), E-selectin, C-X-C Motif Chemokine Ligand 16 (CXCL-16), endothelial NOS (eNOS), and C-X-C motif chemokine ligand 8 (IL-8/CLCX8) in these cells. Those factors are important angiogenesis regulators. Wounds in the feet of diabetic rats had a significantly reduced ulcerated area when treated with EVs from adipose-derived MSCs overexpressing nuclear factor erythroid 2–related factor 2 (Nrf2), a basic leucine zipper (bZIP) protein that regulates the expression of antioxidant proteins. Increased granulation tissue formation, angiogenesis, and levels of growth factor expression, as well as reduced levels of inflammation and oxidative stress-related proteins, were detected in wound beds

Another observation is that my weight loss from the keto diet has ceased, and I feel much more full and satisfied with my diet. This may just be part of the ongoing process of adaptation to fat metabolism (started in Nov '18), but it may also be due to the boost in PPAR activation from taking pioglitazone. The motivation for taking pioglitazone was for adipocyte generation (in addition to the endothelial progenitors generated by statins), but it may also have other benefits.

Observations from phase 2 (complete): Skin continued to look good; I did however experience 2 or 3 mild bouts of cramp in one foot, which may be down to the statins. Looking back at my notes from November 2018, my previous dose of atorvastatin was 5mg/day; this time it was 10mg/day in line with the Janic study. So this might indicate the level at which (for me) side effects starts to occur. I did take Q10.

Observations from phase 3 (in progress): nothing to report.

Other general observations:

1. Blood pressure remained in the normal range. My weight has stabilized after a few months of it falling from keto. I have a blood draw in a few weeks to look at various parameters, I will report back if anything shows up.

2. My meditation has been brilliant recently. I've no idea if this is simply natural progression or due to the protocol. I seem to be able to reliably get into a deep state of aware rest, and finish each practice warm, vascular and restored.

3. I've recently started to beat my personal bests on weight lifting on bench press and deadlift. I recently picked up an injury, however.

Edited by QuestforLife, 07 February 2019 - 10:27 AM.

Back to Sirtuin 4!

I haven’t forgotten about the good old sirtuins and my previous efforts to induce rejuvenation using resveratrol. Recently many pieces of the puzzle have started to fall into place for me – SIRT4, mitochondrial fusion, fatty acid metabolism and the NAD+/NADH ratio. I now feel ready to come up with a new protocol to leverage this new understanding. I am not yet ready to try it out, but I will!

Let’s look at a typically day when I might try and stimulate telomerase using these methods.
In all the following, bear in mind I eat a keto diet and have been since Nov 2018.

Many of the references given below have been more fully explored on the first page of this thread.

Morning
2 tabs of Allicin Max.
Rationale: Use Hydrogen Sulphide to Increase NAMPT (by unknown mechanism).
Reference: https://www.ncbi.nlm...pubmed/27732642
2 sublingual tabs of NMN
Rationale: increase NAD+ without requiring NAMPT to convert NAM.
Coffee and butter
Rationale: Reduce DNA damage to reduce PARP and hence increase the NAD+ pool available to the Sirtuins. Butter in coffee is good for activating ketones, which increase NAD+ and fatty acid metabolism. Fat metabolism is regulated by Sirtuin 4.
References: https://www.ncbi.nlm...pubmed/26632023
Breakfast: keto – bacon, eggs, mackerel
Rationale: Keto raises NAD+/NADH ratio in brain; upregulates fatty acid oxidation and mitochondrial fusion (therefore Sirtuin4)

Resveratrol (dose TBD) in high fat Greek yogurt.
Rationale: Activate Sirtuins (Sirtuin 4), activate telomerase via NAMPT and SIRT4
References: https://www.ncbi.nlm...pubmed/25926556
Afternoon
1 allicin max tab
1 sublingual tab of NMN
Coffee
Lunch: Steak
Evening
1 tab allicin max
1 sublingual tabs of NMN
Dinner: Keto – bacon, eggs, etc
Resveratrol in high fat greek yogurt
90% Dark hot chocolate with stearic acid
Rationale: Boost fatty acid metabolism, mitochondrial fusion and Sirtuin 4
References: https://www.nature.c...467-018-05614-6

Interestingly, we found that serum long chain acylcarnitine levels increase significantly after a 2-day low-fat vegan diet (“0h” vs “basal”, Fig. 3a, b). Conversely, long chain acylcarnitine levels dropped significantly after ingesting the C18:0 drink but not the mock drink (Fig. 3a, b), suggesting that C18:0 intake increases fatty acid beta-oxidation in vivo. Particularly striking, C18:0 ingestion leads to an increase in circulating C18:0-TAG levels (Fig. 2), but causes a decrease in circulating C18:0 acylcarnitine levels (Fig. 3a), indicating that the acylcarnitine drop is not due to reduced substrate availability but rather to increased acylcarnitine usage.

Sirt4 knockout flies have a short lifespan, with increased sensitivity to starvation

Overall, we propose that the SIRT4-OPA1 axis is causally linked to mitochondrial dysfunction and altered mitochondrial dynamics that translates into aging-associated decreased mitophagy based on an unbalanced mitochondrial fusion/fission cycle.

Conclusion: This protocol is aiming to boost NAMPT, the NAD+/NADH ratio, Sirtuin 4 and telomerase.

Comments welcome!

Back to Sirtuin 4!

 

I haven’t forgotten about the good old sirtuins and my previous efforts to induce rejuvenation using resveratrol. Recently many pieces of the puzzle have started to fall into place for me – SIRT4, mitochondrial fusion, fatty acid metabolism and the NAD+/NADH ratio. I now feel ready to come up with a new protocol to leverage this new understanding. I am not yet ready to try it out, but I will!

Let’s look at a typically day when I might try and stimulate telomerase using these methods.
In all the following, bear in mind I eat a keto diet and have been since Nov 2018.

Many of the references given below have been more fully explored on the first page of this thread.

Morning
2 tabs of Allicin Max.
Rationale: Use Hydrogen Sulphide to Increase NAMPT (by unknown mechanism).
Reference: https://www.ncbi.nlm...pubmed/27732642
2 sublingual tabs of NMN
Rationale: increase NAD+ without requiring NAMPT to convert NAM.
Coffee and butter
Rationale: Reduce DNA damage to reduce PARP and hence increase the NAD+ pool available to the Sirtuins. Butter in coffee is good for activating ketones, which increase NAD+ and fatty acid metabolism. Fat metabolism is regulated by Sirtuin 4.
References: https://www.ncbi.nlm...pubmed/26632023
Breakfast: keto – bacon, eggs, mackerel
Rationale: Keto raises NAD+/NADH ratio in brain; upregulates fatty acid oxidation and mitochondrial fusion (therefore Sirtuin4)

Resveratrol (dose TBD) in high fat Greek yogurt.
Rationale: Activate Sirtuins (Sirtuin 4), activate telomerase via NAMPT and SIRT4
References: https://www.ncbi.nlm...pubmed/25926556
Afternoon
1 allicin max tab
1 sublingual tab of NMN
Coffee
Lunch: Steak
Evening
1 tab allicin max
1 sublingual tabs of NMN
Dinner: Keto – bacon, eggs, etc
Resveratrol in high fat greek yogurt
90% Dark hot chocolate with stearic acid
Rationale: Boost fatty acid metabolism, mitochondrial fusion and Sirtuin 4

The protocol is still being tweaked by me, but early indications are interesting. I have increased energy, better mood (problem on keto), more regular bowel movements (problem on keto), decreased appetite, decreased desire for caffeine and possibly better sleep (was already good).

As it stands, this is what I'm taking:

AM and PM 

Resveratrol and Curcumin in yogurt ( approx. 400mg of each; still tailoring this) - for Sirtuins and Nrf2

Allicin Max for hydrogen sulphide (1 tab = 180mg stabilised allicin)

NMN for NAD+ (2 sublingual tabs = 250mg)

Trimethyl glycine (2-3 g) for methylation (impedes blocking of Sirtuins by NAM) - don't take this every time

AM only 

Cyclo-astragenol (25mg)

I've dropped any midday dosing as I think it's a good idea to let the body have some sort of homeostasis and do what it needs to do during periods of lower NAD+ and higher ROS, etc.

I'm still drinking coffee to reduce DNA damage (although I don't feel the need so much, anymore) and I'm still have a dark chocolate and stearic acid hot chocolate in the evening. Sill also eating keto. 

Very interesting:

https://www.ncbi.nlm...les/PMC4547093/

Turns out SIRT4 is required to help telomerase bond to the telomere.

This explains, in part, the telomere lengthening ability of resveratrol, as reported up-thread. I'm still not clear on how telomerase itself is upregulated, but I suppose SIRT4 allows access to HTERT (human telomerase gene) is a similar way to how it allows telomerase to bind to the telomere and lengthen it. This might sound far-fetched but we know that there are sections of telomere within the chromosome, including some quite close to HTERT; at those locations SIRT4 could also bind.

I am not clear how this ties in with SIRT4 up-regulation of fatty acid metabolism via the OPA1 protein and mitochondrial fusion. It certainly tallies with the healthy nature of ketogenic diets (is there data on telomere length for those on keto?), but it also holds a note of warning – constant mitochondrial fusion will suppress mitophagy and eventually lead to a rise in ROS, which is another pathway to senescence.

Maybe, just maybe, this is a double whammy aging trap – lengthen telomeres but get higher ROS into the bargain, or allow telomeres to shorten. Another way of looking at it is that under starvation the body changes to burning it's fat reserves, and under those conditions also gets access to more telomerase to help get it through the hard times. As soon as plentiful glucose access is re-established however, normal controls on telomerase are also restored.

Edited by QuestforLife, 18 February 2019 - 04:30 PM.

AM and PM 

Resveratrol and Curcumin in yogurt ( approx. 400mg of each; still tailoring this) - for Sirtuins and Nrf2

Allicin Max for hydrogen sulphide (1 tab = 180mg stabilised allicin)

NMN for NAD+ (2 sublingual tabs = 250mg)

Trimethyl glycine (2-3 g) for methylation (impedes blocking of Sirtuins by NAM) - don't take this every time

 

AM only 

Cyclo-astragenol (25mg)

I added pterostilbene to the resveratrol + curcumin in yogurt, but after a few doses it became too much - felt pretty bad; lethargic and weak. So I'm dropping SIRTUIN activators for a while.

I'm going to concentrate on raising NAD+ and NAMPT instead, and let the SIRTUINS and NRF2 take care of themselves as required - I figure being on Keto will activate SIRTUINS and NRF2 anyway, so less need to add activators (might re-introduce them on a cyclical basis in the future).

One concern I have with long term keto is the raised SIRTUIN4 and OPA-1 protein leading to constantly upregulated mitochondrial fusion and downregulated mitophagy. I'm not sure how this fits in with the widely reported improved mitohormesis associated with keto (higher short term ROS, lower long term ROS) - but I'm not taking any chances and intend to upregulate mitophagy aggressively with Nicotinamide starting today.

I'm combining Nicotinamide (3g) with Taurine (3g) and NMN (250mg) for this purpose.

One interesting recent finding is that taking taurine (along with my usual allicin Max) seemed to stop the usual tiredness I experience from the fusion upregulating Stearic Acid. So it might be that the resultant rise in NAMPT (via Hydrogen sulphide) is supporting greater recycling of NAM (nicotinamide) in NAD+. I'm not clear on whether the raised NAD+ is counteracting fusion via mitophagy (it would have to be very high to do this), or whether it is some other effect.

One interesting recent finding is that taking taurine (along with my usual allicin Max) seemed to stop the usual tiredness I experience from the fusion upregulating Stearic Acid.

Solved!

https://en.wikipedia...etobutyric_acid

Ketobutyrate is one of the break down products of threonine (taken as part of Turnbuckle's Stem Cell Protocol with the stearic acid) and is also an enzyme used by the body to make Hydrogen Sulphide. See:

https://openheart.bm...ent/4/1/e000600

So taking taurine (also boosts H2S) probably makes the threonine go further.

Also interesting is the fact that further downstream metabolism of ketobutyrate in the mitochondria uses biotin as a co-factor, which explains why after 3 months in keto I've had to start supplementing biotin to avoid drying out!

 

One concern I have with long term keto is the raised SIRTUIN4 and OPA-1 protein leading to constantly upregulated mitochondrial fusion and downregulated mitophagy. I'm not sure how this fits in with the widely reported improved mitohormesis associated with keto (higher short term ROS, lower long term ROS) - but I'm not taking any chances and intend to upregulate mitophagy aggressively with Nicotinamide starting today.

 

I'm combining Nicotinamide (3g) with Taurine (3g) and NMN (250mg) for this purpose.

 

 

Update: I did 3 days of mitochondrial fission.

Day 1 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning  evening.

Day 2 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning only.   I took 250mg of NMN in the evening

Day 3 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning only.   I took 250mg of NMN and 2g of taurine in the evening.

I didn't feel weak aside from a slight feeling of shakiness after the first dose. By day 3 I definitely felt a little strange though (very hard to describe aside from a feeling of mental lethargy and diminished sex drive without physical tiredness. It is similar to what prolonged resveratrol supplementation does to me) and that feeling came back with a vengeance when I took the taurine on the evening of the 3rd day even though I didn't take nicotinamide (though no doubt there was plenty left in my system).

Since then I have discontinued nicotinamide and taurine but continued with NMN.

My new dosing regime is 125mg 4 times a day, with an allicin supplement morning and evening. Interestingly this seems to have a greater effect than when I've used 250mg NMN twice a day (perhaps it's to do with the sublingual tablets). I've had periods of unexpected fatigue interspersed with periods of what I can only term hyper-focus. It's similar to some of the positive effects I got from resveratrol after supplementing it for months, when I wasn't really paying attention to what was going on (say when driving), but without even trying I can see everything with ease and nothing can surprise me. I almost feel like I could reach out and pluck a bird from the wing. Weird feeling.

So I'm not sure what's going on. I did Turnbuckle's mitochondrial fission protocol many times in the past, but not for a over a year. I was worried that keto and repeated bouts of the fusion protocol may have decreased the quality of my mitochondria, but that doesn't seem to be the case given it took 3 days of heavy nicotinamide use to get much reaction. Still NMN does appear to be doing something.

Another interesting study on SIRT4

https://www.research...A_Decarboxylase

All adding to the picture that SIRT4 is absolutely essential in the metabolism or genesis of lipids depending on whether it is turned on or off - and seems to support my assertion that a low carb, high fat diet would support life extension.

I'm adding this picture of the melvonate pathway for future reference, as I believe it will be a great starting reference to see how various interventions including Keto diet, statins, resveratrol, etc. effect mitochondria and from there the nuclear DNA and telomeres.

Edited by QuestforLife, 27 February 2019 - 03:22 PM.

Update: I did 3 days of mitochondrial fission.

 

Day 1 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning  evening.

 

  Did you take NAM without Ribose?

  P.S. I also tried sublingual NMN and its a different beast from NAM. I also experienced periods of `unintended` focus, it's probably similar to focus that I got from deep ketosis but kinda ramped up version of it.

When I started a ketogenic diet I was rarely lower than 3mmol so I enjoyed it quite a bit, now, unfortunately, I get it only while fasting so NMN is a good back door for it)

Did you take NAM without Ribose?

P.S. I also tried sublingual NMN and its a different beast from NAM. I also experienced periods of `unintended` focus, it's probably similar to focus that I got from deep ketosis but kinda ramped up version of it.
When I started a ketogenic diet I was rarely lower than 3mmol so I enjoyed it quite a bit, now, unfortunately, I get it only while fasting so NMN is a good back door for it)

High Desert Wizard brough to my attention the effects on bee keeping and honey on telomeres, see this paper where beekeepers had significantly longer telomeres than the age matched (actually younger) control group:

 

https://www.ncbi.nlm...rticle_9797.pdf

 

Interestingly royalactin (the most common protein in royal jelly and the one responsible for differentiating queen bees from drones ) has been shown to maintain stemness, at least in vitro:

 

https://www.nature.c...467-018-06256-4

 

Very rough calculation, this would take ATLEAST 15g of royal jelly a day to replicate the concentration in vivo, so I'm not certain I'm willing to take that risk to add it to my protocol just yet (only seen a human study where they took up to 3g a day).

Friend of mine was gobbling RJ by the kilo. He got it in bulk off the internet, shipped frozen. He LOVED it, said it was an integral part of his Lyme disease recovery program. 

 

https://www.ncbi.nlm...pubmed/24882731

Long-term administration of Greek Royal Jelly improves spatial memory and influences the concentration of brain neurotransmitters in naturally aged Wistar male rats.

So take that for what it is. 

Update: I did 3 days of mitochondrial fission.

 

Day 1 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning  evening.

Day 2 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning only.   I took 250mg of NMN in the evening

Day 3 I took 3g of nicotinamide, 3 g of taurine and 250mg of NMN in the morning only.   I took 250mg of NMN and 2g of taurine in the evening.

 

I didn't feel weak aside from a slight feeling of shakiness after the first dose. By day 3 I definitely felt a little strange though (very hard to describe aside from a feeling of mental lethargy and diminished sex drive without physical tiredness. It is similar to what prolonged resveratrol supplementation does to me) and that feeling came back with a vengeance when I took the taurine on the evening of the 3rd day even though I didn't take nicotinamide (though no doubt there was plenty left in my system).

 

Since then I have discontinued nicotinamide and taurine but continued with NMN.

 

My new dosing regime is 125mg 4 times a day, with an allicin supplement morning and evening. Interestingly this seems to have a greater effect than when I've used 250mg NMN twice a day (perhaps it's to do with the sublingual tablets). I've had periods of unexpected fatigue interspersed with periods of what I can only term hyper-focus. It's similar to some of the positive effects I got from resveratrol after supplementing it for months, when I wasn't really paying attention to what was going on (say when driving), but without even trying I can see everything with ease and nothing can surprise me. I almost feel like I could reach out and pluck a bird from the wing. Weird feeling.

 

So I'm not sure what's going on. I did Turnbuckle's mitochondrial fission protocol many times in the past, but not for a over a year. I was worried that keto and repeated bouts of the fusion protocol may have decreased the quality of my mitochondria, but that doesn't seem to be the case given it took 3 days of heavy nicotinamide use to get much reaction. Still NMN does appear to be doing something.

For what it's worth, perhaps my experience may shed light on your issues with that fission combo. My experience is that NAM is the problematic ingredient, not taurine, not NMN. I've lost count of the number of times before cycling for a couple hours, I've taken 250 mg powder sublingual NMN, then in my hydration bottle with some juice/H2O - 5 g Ribose, 2-4 g taurine, followed during the ride with 500-750 mgs sublingual MNM tabs. During or after the ride, no issues, no problems, great stamina.

However, I've did a combo once including NAM in doses like you have, and I find I didn't feel right, or particularly good. Maybe it's just me, but after a couple days I stopped and don't take NAM anymore in any large amount. Allicin, on the other hand, seems to be one of the best supps I've ever started taking, with (what seems like) most noticeable effects on joints, knees in particular, and hopefully is synergistic with my daily NMN as well.

Edited by Oakman, 27 February 2019 - 07:27 PM.

Friend of mine was gobbling RJ by the kilo. He got it in bulk off the internet, shipped frozen. He LOVED it, said it was an integral part of his Lyme disease recovery program.

So take that for what it is.

Edited by QuestforLife, 28 February 2019 - 12:32 PM.

For what it's worth, perhaps my experience may shed light on your issues with that fission combo. My experience is that NAM is the problematic ingredient, not taurine, not NMN. I've lost count of the number of times before cycling for a couple hours, I've taken 250 mg powder sublingual NMN, then in my hydration bottle with some juice/H2O - 5 g Ribose, 2-4 g taurine, followed during the ride with 500-750 mgs sublingual MNM tabs. During or after the ride, no issues, no problems, great stamina.

However, I've did a combo once including NAM in doses like you have, and I find I didn't feel right, or particularly good. Maybe it's just me, but after a couple days I stopped and don't take NAM anymore in any large amount. Allicin, on the other hand, seems to be one of the best supps I've ever started taking, with (what seems like) most noticeable effects on joints, knees in particular, and hopefully is synergistic with my daily NMN as well.

Agreed Oakman, there is something special about NMN. Perhaps it can have the beneficial effects of mitophagy and sirtuin activation at a much lower dose than nicotinamide.

Also agreed about allicin; definitely a keeper in my supplements list - especially if you believe that microbial overgrowth has a contribution to aging, like I do.

My concern about allicin is its effect on the microbiome. Is it killing off beneficial bacteria? Or is it altering the microbiome - due to its natural antibiotic effect - in a negative way? Who knows? 

https://www.quora.co...od-gut-bacteria

My concern about allicin is its effect on the microbiome. Is it killing off beneficial bacteria? Or is it altering the microbiome - due to its natural antibiotic effect - in a negative way? Who knows? 

 

https://www.quora.co...od-gut-bacteria

Do a bit of Googling on this and it should put your fears to rest. For example, Garlic compounds may boost cardio health indirectly via gut health

Another interesting study on SIRT4

 

https://www.research...A_Decarboxylase

 

All adding to the picture that SIRT4 is absolutely essential in the metabolism or genesis of lipids depending on whether it is turned on or off - and seems to support my assertion that a low carb, high fat diet would support life extension.

Having read the complete paper now, this it a bit of a head-scratcher.

Turns out SIRT4 does exactly the opposite to what I had assumed. SIRT4 activation causes lipogenesis and inhibits fat oxidation. This makes absolutely no sense, given we know SIRT4 is activated in periods of starvation (for example mTOR inhibition can increase SIRT4 via CREB, see https://www.ncbi.nlm...es/PMC3684628/ ).

So why would starvation cause the body to make fats and not burn them? It wouldn't, of course. But the paper is quite rigorous, even showing:

...SIRT4 KO mice display deregulated lipid metabolism, leading to increased exercise tolerance and protection against diet-induced obesity.

And this isn't an isolated result from just one paper. Here (https://www.ncbi.nlm...pubmed/29081403) we see that SIRT4 associates with OPA1 (mitochondrial fusion protein) and that this can impair mitophagy. This paper (https://www.ncbi.nlm...pubmed/28758339) finds the same thing.

This paper (http://diabetes.diab...lement_1/276-LB) confirms the OPA1-SIRT4 link beyond doubt by finding that mice lacking OPA1 were immune to high fat induced obesity.

We generated mice lacking OPA1 in adipose tissue (OPA1 Ad-KO) and demonstrated that OPA1 deletion in adipose tissue completely prevents diet-induced obesity (DIO).

After thinking about this for a few hours, the only conclusion I can come to, is that this is one of the proteins that does different things depending on where it is located. We know from the yeast study I posted up thread (https://www.ncbi.nlm...pubmed/26218225) that:

Sir4 ... is required for Ku-mediated telomere lengthening and telomerase recruitment

For this SIRT4 would need to be located in the nucleus. And yet all the effects we are seeing on OPA1 and mitochondrial fusion are occurring in the mitochondria. So my hypothesis is as follows: in a nutrient-rich state SIRT4 locates to the mitochondria, causes fusion via OPA1 in order to minimise mitophagy and maximise ATP production. But in nutrient-starvation state (high NAD+/NADH) or during a period of mitophagy (for example triggered exogenously by resveratrol), mitochondria are broken up and autophagy is up-regulated and SIRT4 is released from the mitochondria. SIRT4 will also be produced from the nucleus under starvation. Therefore in this state SIRT4 will be where it needs to be to elongate telomeres while at the same time reducing OPA1 and causing elevated quality control in the mitochondria.

This is only a hypothesis, but it fits all the facts and I will be doing further research to strengthen the argument further. I also need to fill in the blanks to explain the effects of statins and stearic acid on mitochondria, which are related.

 

For this SIRT4 would need to be located in the nucleus. And yet all the effects we are seeing on OPA1 and mitochondrial fusion are occurring in the mitochondria. So my hypothesis is as follows: in a nutrient-rich state SIRT4 locates to the mitochondria, causes fusion via OPA1 in order to minimise mitophagy and maximise ATP production. But in nutrient-starvation state (high NAD+/NADH) or during a period of mitophagy (for example triggered exogenously by resveratrol), mitochondria are broken up and autophagy is up-regulated and SIRT4 is released from the mitochondria. SIRT4 will also be produced from the nucleus under starvation. Therefore in this state SIRT4 will be where it needs to be to elongate telomeres while at the same time reducing OPA1 and causing elevated quality control in the mitochondria.

 

This is only a hypothesis, but it fits all the facts and I will be doing further research to strengthen the argument further. I also need to fill in the blanks to explain the effects of statins and stearic acid on mitochondria, which are related.

This also raises the question of whether a ketogenic diet is likely to result in mitochondrial fusion, and the consequent boost to ATP production and lipogenesis (fat storing). It seems obvious now that this will not be the case, and that the opposite is likely - keto is likely to be similar to starvation in that low insulin levels will suppress mTOR and raise SIRT4 expression levels (apart from around meal times). Similarly OPA1 is likely to be repressed and mitochondria fission and mitophagy increased, and this jives with what we know of a ketogenic diet protecting from cancer and aging.  

This is different to the case of statins, where lipogenesis is blocked, but OPA1 is not - in fact it has been shown that this is raised (see https://www.ncbi.nlm...bmed/27720611),which leads to mitochondrial fusion. And this potentially explains how statins can lead to de-differentiation (via GTPases, and also mitochondrial fusion - same as Stearic acid) and maybe longer telomeres (higher SIRT4 through interaction with OPA1, though it's not clear how these then get located in the nucleus). However the downregulated mitophagy would eventually lead to dysfunctional mitochondria (as is widely reported by long term users).

Stearic acid upregulates mitochondrial fusion via the strong signal of fatty acids ready to be metabolised.