1033 replies to this topic

[Andey]

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.

  

  Looks like short life statins like pravastatin would be the best.

  How would statins work combined with a ketogenic diet?

 

  P.S. I haven't found studies where insulin regulates mTor, looks like that link work in the opposite direction - mTor inhibition downregulates insulin secretion

Edited by Andey, 04 March 2019 - 02:45 PM.

[QuestforLife]

Looks like short life statins like pravastatin would be the best.
  How would statins work combined with a ketogenic diet?

Well according to Janic et al, a low dose statin is best, see https://www.ncbi.nlm...pubmed/28951255

Also they found an intermittent rather than chronic dosing is superior with peak benefit in rats at ~6weeks, see: https://www.ncbi.nlm...pubmed/27010807

When I tried out atorvastatin and losartan I found I had better results dosing for 1 month (5mg atorvastatin, 25mg losartan) than I did when dosing for 2 weeks (10mg atorvastatin, 25mg losartan, 30mg pioglitazone). On both occasions I was on Keto, although the latter time was Jan (3 + months keto), the former time I had only just started keto (November).

I'm unsure exactly about the utility of statins + keto, but I still think the ROCK inhibition effects of statins make them a worthwhile addition to use a few times a year. I might have more to say when I've done more research (story of my life!).

 

P.S. I haven't found studies where insulin regulates mTor, looks like that link work in the opposite direction - mTor inhibition downregulates insulin secretion

You might be right, although I have a suspicion the link goes both ways. Even if it doesn't, insulin is a powerful growth hormone in its own right; i'd argue stronger even than mTOR - not pound for pound of course, but over the course of 24 hours - compare the state of someone constantly snacking on carbs (normal western diet) to someone who occasionally eats a meat heavy meal (keto), and I think we'd agree who is anabolic and who is catabolic!

[QuestforLife]

I found a very instructive paper

 

https://www.cell.com...15?showall=true

 

 

Highlights

• Peridroplet mitochondria (PDM) have enhanced bioenergetic capacity
• Peridroplet mitochondria have low fatty acid oxidation capacity
• PDM support lipid droplet expansion by providing ATP for triacylglyceride synthesis
• Distinct fusion-fission dynamics separate PDM from cytoplasmic mitochondria

 

Turns out there is a distinct type of mitochondria that fuse around droplets of fat and enlarge them. It now becomes clear that this is the mechanism by which SIRT4 and OPA1 work together to trigger lipogenesis (and store fats for hard times). It explains a great many things:

• How Flies lacking Sirt4 could not withstand starvation (it was not from a lack of ability to burn fats, but to store them in the first place) - see reference up thread
• How David Sinclair's classic experiments feeding high fat diet induced obese mice resveratrol did not make them thin (it just made them store fat more!), but did keep them healthy (possibly via SIRT1 and mitophagy)
• How trout with a diet very low in fish oils, still managed to be high in EPA and DHA if fed resveratrol (see DOI: 10.3390/md15080252)

So from all this I can now conclude that the mechanism by which Resveratrol generates lipids via SIRT4 and induces mitophagy (via SIRT1) are distinct and co-incidental. It appears SIRT4 is actually activated not in starvation induced conditions, but in the opposite case (and therefore probably not in keto). It is triggered to store fat for future hard times.

 

This doesn't get us any closer to knowing how this is linked to telomerase activation (posted at the top of the thread, or here: 'A critical role of nicotinamide phosphoribosyltransferase in human telomerase reverse transcriptase induction by resveratrol in aortic smooth muscle cells' https://www.ncbi.nlm...bmed/25926556),but there are some more interesting clues in how SIRT4 is both activated and inhibited.  

 

See 'Crystal structures of the mitochondrial deacylase Sirtuin 4 reveal isoform-specific acyl recognition and regulation features'

 

https://www.nature.c...467-017-01701-2

 

 

 

NAM appears to act as a physiological regulator for most sirtuin isoforms37, and a NAM dose-response experiment with Sirt4 revealed potent inhibition with IC 50 = 13 ± 2 μM (Fig. 4d). The effect on Sirt4 is even more potent than on other isoforms37,46 and possibly supported by the Sirt4-loop at the NAM accommodating C-site entrance. An NADH dose-response experiment (Fig. 4e), corrected through NADH spiking controls for its fluorescence overlap with AMC47, also indicated pronounced Sirt4 inhibition (IC 50 = 126 ± 12 μM at 500 μM NAD+). Analyzing the NADH titration with the robust MS assay confirmed this Sirt4 inhibition potency (IC 50 = 142 ± 54 μM; Supplementary Fig. 4b) that exceeds NADH effects on other sirtuins (IC 50 1.3–27.9 mM)47,48, consistent with Sirt4 nucleotide site features (see above and discussion). The response of Sirt4 activity to NADH levels around ~30 μM, which is estimated to be the mitochondrial concentration of free NADH47, suggests NADH or the NAD+/NADH ratio to act as a physiological Sirt4 regulator.

 

This is saying that NAM inhibits SIRT4 at much lower concentrations that the other SIRTUINS, and that NADH or a low NAD+/NADH ratio may also inhibit SIRT4.

 

This adds a lot of weight to why in 'A critical role of nicotinamide phosphoribosyltransferase in human telomerase reverse transcriptase induction by resveratrol in aortic smooth muscle cells', NAMPT was critical. At the time I thought it was because SIRT4 (like all SIRTUINS) needed NAD+, but now I see that it is NAM that needs to be kept at low concentrations (conversion to NAD+ by NAMPT).

 

This also shreds new light on my original report.

 

 

 

I took 2 drops (~1mg) of Methylene blue for the first time in 2 months. This was in addition to my normal AM supplements that include Allicin, Vitamin C and Ashwaghanda. Within 4 hours I felt unusually fatigued and had a headache. The experience was similar to using Turnbuckle’s mitochondrial fission Nicotinamide and Ribose protocol for the first time (https://www.longecit...drial-dynamics/ ), but which has subsequently and even recently failed to elicit the same effect, even when combined with the same supplements. I have not experienced this fatigue when using MB alone.
As my headache grew worse I decided to try and trigger mitochondrial fusion by ingesting 20g of fish oil (ref: https://www.ncbi.nlm...les/PMC3963938/ ). I gave it two hours to work but the effect was minimal. I lay down for a rest. Letting my mind wonder over various papers I’d read recently the potential synergy of resveratrol with high NAMPT/NAD occurred to me. I took 10ml (180mg) of liposomal resveratrol from Actinovo that I had lying about. This liposomal resveratrol supplement had not previously produced any significant positive results, and I had taken it at various doses from 180mg to 4 g a day.
Within 10 minutes not only did I feel better, but I felt euphoric. I ran down the stairs and played with my kids in the garden with a renewed and fresh energy. I continued to feel fantastic for the rest of the day.

 

Notice that I took  Methylene blue (competes with the electron transport chain converting NADH to NAD+), then Fish Oil (likely to be used in a nutrient replete state to make fat stores), then resveratrol (upregulated SIRT4 and NAMPT).

 

It's all here.

 

 

 

 

Edited by QuestforLife, 06 March 2019 - 12:11 PM.

[QuestforLife]

Some other insights based on the above, briefly.

 

I've already posted on fish oil, but the above post does shed additional light on why this would upregulate mitochondrial fusion (for example here: https://doi.org/10.3...ys.2015.00109),specifically in relation to lipogenesis.

 

Statins is an interesting case; they're known to lead to mitochondrial fusion (for example here: https://doi.org/10.1...l.2016.09.004),and I expect the reason is that they block the melvonate pathway (see the diagram I put up on the last page). The melvonate pathway creates cholesterol amongst other things, but competes with melonyl CoA for the first step in creating acetyl CoA. If the conversion from melonyl CoA to acetyl CoA is impeded (i.e. via SIRT4), then lipogenesis is increased. So it stands to reason than in inhibiting the melvonate pathway further down, Statins are achieving something similar. It's all speculation at this point.

 

Finally stearic acid and mitochondrial fusion. In the human study on stearic acid ingestion that famously caused mitochondrial fusion in neutrophils (DOI: 10.1038/s41467-018-05614-6), consumption of stearic acid seemed to upregulate fatty acid oxidation, as detected by a reduction in circulating acyl carnitine levels (fatty acids ready to be burned). This is the opposite to what we'd expect. These carnitine levels rose after 2 days of vegan (low fat) diet, so it might well be that abundant carbohydrates are upregulating fat storage, which Stearic acid is then triggering the burning of. So it might be that the mitochondrial fusion triggered by stearic acid is distinct from the mitochondrial fusion triggered by SIRT4 and OPA1, which is done for the purposes of creating fat storage not burning it. This agrees with the fact that stearic acid consumption appears to be healthy, whereas prolonged SIRT4-OPA1 activation is not (because it results in fat accumulation and possibly down-regulation of mitophagy).

 

Based on the (at this time) speculative arguments above, statins would not combine well with keto, but stearic acid would. It also suggests that the addition of pioglitazone - a known PPAR (fat burning) agonist - to my last statin-sartan cycle, might have been a mistake.

 

 

Edited by QuestforLife, 06 March 2019 - 02:45 PM.

[QuestforLife]

I have a hypothesis that resolves a few issues.

 

Several papers I have read have attempted to cast SIRT4 in a bad light, i.e. promoting aging. For example 'SIRT4 interacts with OPA1 and regulates mitochondrial quality control and mitophagy' (reference link up-thread) talks about inhibition of mitophagy. And more recently I've read the damning 'MicroRNA-15b regulates mitochondrial ROS production and the senescence-associated secretory phenotype through sirtuin 4/SIRT4' (https://www.ncbi.nlm...les/PMC4833141/), which discusses how SIRT4 is upregulated in a models of cellular senescence and photo damaged skin.

How can this be the case?

 

I think this is a clear example of putting the cart before the horse. Hormetic responses such as the sirtuins occur to counteract stress, and end up being beneficial – they don’t cause damage. If we go back to 'SIRT4 interacts with OPA1 and regulates mitochondrial quality control and mitophagy' we see that one of the ways they upregulate SIRT4 and mitochondrial fusion is through exposure to ionizing radiation (it's very effective). Clearly in that case it is the radiation that is causing the damage, and SIRT4 that responds. We also know from 'A critical role of nicotinamide phosphoribosyltransferase in human telomerase reverse transcriptase induction by resveratrol in aortic smooth muscle cells' that resveratrol upregulates SIRT4 very effectively. It's not much of a leap to deduce that it does this via mitochondrial fission. Fig 3 from the 'Nicotinamide-inducedMitophagy..' paper (attached) shows that resveratrol is the most effective at inducing punctae in mitochondria from the tested compounds (including nicotinamide). Again it is the ‘toxin’ resveratrol ca\using the damage, and SIRT4 that responds.

 

This makes total sense to me. From an evolutionary point of view, we know flies lacking SIRT4 die quickly from starvation compared to wild type controls, (explained up-thread by an inability to store fat without SIRT4). And we know SIRT4-OPA1 mitochondria are resistant to normal fission-fusion cycles. Now we know why! Stress – like ionizing radiation, stilbenoids, flavonoids, etc. – activate the sirtuins. Now in the mitochondria responsible for lipogenesis, SIRT4 causes the mitochondria to resist fission and mitophagy. This enables, for example, flies that are stressed to store fats to survive a subsequent period of drought or starvation; SIRT4 knockout flies cannot and die swiftly.

Attached Files

•  RES induced Mitophagy PIC.pdf   2.32MB   398 downloads

[QuestforLife]

 

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.

 

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.

 

The next iteration of my statin-sartan treatment is starting again next week with some modifications:

 

1. Phase 1 is the Senolytics phase, but this time I will be replacing the senolytic with a high (for me) dose of rapamycin, based on this paper: 'Reprogramming glioblastoma multiforme cells into neurons by protein kinase inhibitors' (https://link.springe...3046-018-0857-5), which showed a combination of mTOR and ROCK inhibitors was most effective in de-differentiating glioblastoma (cancer) cells back into neural progenitors.

 

Cellular senescence has been shown to regulate reprogramming fibroblasts to iPS cells and fibroblast-neuron conversion. Since many protein kinases are involved in senescence and proliferation processes, we screened a protein kinase inhibitor library . ...Eight candidate small molecules/compounds were selected for further confirmation. Secondary screening confirmed that three compounds induced morphological changes with higher efficiency. Two candidate small molecules, namely rapamycin and palomid 529 (P529), are mTOR inhibitors and the third one (Y27632) is a ROCK inhibitor. See attached figure

 

Although my protocol is not aimed at being anti-cancer, I am hoping I'll generate more progenitors in general by including a mTOR inhibitor. This will be a single dose.

 

I may in a future iteration re-adopt senolytics, once I have identified a more effective senolytic protocol.

 

2. Phase 2 is progenitor cell generation, currently 14 days of 10mg atorvastatin and 25mg losartan daily. I have removed pioglitazone from this stage.  Phase 2 will begin on the same day as Phase 1.

 

3. Phase 3 is 21 days of telomere stimulation. This will start in week 2 and consist of 20mg of (nano-) cycloastragenol, folate and TMG to help with cell division.

 

4. Phase 4 is a 28 day wash-out/differentiation phase.

 

I am currently trialling a NAD+ boosting protocol (which I will post on separately), but will discontinue it for PHASE 1+2 (and possibly Phase 3).

 

Attached Thumbnails

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Edited by QuestforLife, 12 March 2019 - 10:32 AM.

[QuestforLife]

I am currently trialling a NAD+ boosting protocol (which I will post on separately), but will discontinue it for PHASE 1+2 (and possibly Phase 3).

 

After reading the following paper - 'Restoration of Mitochondrial NAD1 Levels Delays Stem Cell Senescence and Facilitates Reprogramming of Aged Somatic Cells' (https://www.ncbi.nlm...pubmed/27428041) I have reconsidered stopping NAD+ boosting supplements during my statin-sartan protocol.

 

 

Importantly, restoring mitochondrial NAD+ levels by overexpressing NNT and NMNAT3 enhanced reprogramming efficiency of aged somatic cells

 

Although this paper is primarily focussing on generating induced pluripotent stem cells (using OSKM) rather than progenitor cells (using ROCK and mTOR inhibition), it clearly demonstrates that low NAD+ levels can impair reprogramming; hence I have the opportunity to combine the two threads of my investigation by combining the cyclical statin-sartan protocol with my chronic NAD+ boosting protocol.

 

I will post a new combined protocol soon.

Edited by QuestforLife, 13 March 2019 - 10:16 AM.

[Brian Valerie]

Just want to publicly express how much I "like" (and no doubt many of us following this thread feel the same way) your detailed sharing of your knowledge, hypotheses, and self-experimentation experiences with us.  While I don't place tremendous faith in anecdotal evidence, with its great susceptibility to the placebo effect and confounding variables, n=1 is obviously better than n=0. Thanks, QuestforLife!     

[QuestforLife]

Just want to publicly express how much I "like" (and no doubt many of us following this thread feel the same way) your detailed sharing...

Thanks Brian. Just looking back at the thread it's obvious that I've learnt a great deal in the process and changed my mind about how to go about age-reversal several times! Hopefully the process will continue and we'll end up with something that atleast makes a dent in aging.

[QuestforLife]

A short update:

NMN and MB (methylene blue) appear to be synergistic, which isn’t suprising given that MB activates NRF2, reduces ROS and (temporarily) raises NAD+.

 
I’ve been taking 1 mg of MB in the morning along with 1 tab of allicin Max, and sublingual NMN all day (4 tablets split into 2 makes 8 doses of approx. 60mg each separated by atleast 1 hour). I’ve also been drinking green tea, which seems to increase the heightened focus I get from NMN.

I’ve generally been feeling good with a slight increase in overall energy but nothing huge to report. But then I decided to take a further 0.5mg of MB with my last dose of NMN (about 7pm). I did some HIIT (high intensity interval training) an hour later and my performance was ridiculous. I do a pretty intense, all-out 30 secs on, 30 secs rest for 5 minutes punching and kicking the heavy bag. I’ve done this hundreds, maybe thousands of times and this was probably my best performance ever in terms of intensity and lack of fatigue. That night however my sleep was disrupted. I woke up 3 hours early (4am instead of 7am), and didn’t manage to more than doze till morning. Every time I drifted off something would jerk me awake. It felt like I might have an occasional jittery heartbeat. Sleepless nights are extremely unusual for me; in fact I don’t think insomnia like this has ever happened to me before. Surprisingly, I didn’t actually feel tired and was able to operate normally the next day (although the following night I did sleep very well indeed – without an evening dose of MB).

 

I've repeated the HIIT without taking MB (just NMN) in the evening and my performance is not as good. So I've decided to increase the morning dose of MB to 1.5mg.

Edited by QuestforLife, 14 March 2019 - 03:34 PM.

[QuestforLife]

Some more research on SIRT4
I’ve reported on some of these findings before, but recently it all seems to be coming together in my head and I felt the need to summarise it, if only for my own reference. But I think it will be of interest to others, as well.

Turns out glutamine intake causes a metabolic shift away from glucose metabolism [https://www.ncbi.nlm...s/PMC4344121/].Glutamine is the preferred fuel for immune cells and also cancer; it’s loved by fast dividing cells basically, whereas quiescent cells prefer glucose. Turns out that glutamine causes an increase in SIRT4 and it is believed that this response is anti-cancer: SIRT4 increases to block excessive glutamine metabolism. Now we also know SIRT4 is involved in shifting metabolism away from making acetyl-CoA and (eventually) ATP and towards fat storage (energy for later use).  So the general theme is SIRT4 stopping division and ATP production and pausing cell division while storing energy for later. It turns out that this is exactly the way that cellular arrest is accomplished when the DNA damage response (DDR) occurs. In DDR you need to arrest division until the damage is repaired, unless damaged chromosomes get reproduced and this can result in cancer. So DDR activates SIRT4 to block glutamine metabolism and enforce cellular arrest (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4851233/). This is how rapamycin works as well (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3684628/) and the reverse is obviously true – mTOR upregulates glutamine metabolism and represses SIRT4 (same paper).  We can link this back to the damage caused either from resveratrol or ionising radiation, which also upregulate SIRT4 – I thought this damage was just to mitochondria resulting in mitochondrial fission, and maybe it is, but it might be that this fission is a trigger for ROS that then causes DDR and it is this that then upregulates SIRT4 from the nucleus. And we know this then gets back to the mitochondria and links to L-OPA1 (fusion protein) to shift energy to being stored rather than used for ATP (for cell division amongst other things). See how I said it was all starting to link up? I wasn’t kidding! So maybe this is what happens every time we have a big meal? We get lots of energy, cells start dividing, inevitably some DDR occurs and SIRT4 steps in to put on the brakes on and store the energy whilst the necessary repairs take place. But then don’t forget the whole process is dependent on NAD+ being available for SIRT4, and also – SIRT4 is inhibited at lower concentrations of NAM and NADH than the other sirtuins, so it is probably only activated in a transitory fashion.

The growth and division – arrest, store energy and repair axis also tentatively links us back to the  yeast research I highlighted previously when SIR4 (yeast sirtuin) located to the telomeres and helped recruit telomerase, but it competed there with the RAS an RAP proteins, which are pretty much synonymous with growth and proliferation. So maybe (I’m speculating here) telomere elongation (in normal yeast cells, at least) requires for growth to stop. Clearly this is an area where more research is required.

 

So how do we activate SIRT4 more than it would normally be activated in daily life? These are my thoughts so far:
• High NAD+
• Well-fed, no-exercise state
• Rapamycin
• L-Glutamine
• Resveratrol

[QuestforLife]

Another article that supports the above findings that SIRT4 controls growth, here through controlling insulin secretion: https://www.cell.com...9456(17)30193-9

 

 

Active SIRT4 decreases mitochondrial fuel oxidation into the TCA cycle, ultimately resulting in a decreased ATP/ADP ratio to inhibit insulin secretion

 

 

 

 

 

Attached Thumbnails

• 

[marcobjj]

A short update:

NMN and MB (methylene blue) appear to be synergistic, which isn’t suprising given that MB activates NRF2, reduces ROS and (temporarily) raises NAD+.

 
I’ve been taking 1 mg of MB in the morning along with 1 tab of allicin Max, and sublingual NMN all day (4 tablets split into 2 makes 8 doses of approx. 60mg each separated by atleast 1 hour). I’ve also been drinking green tea, which seems to increase the heightened focus I get from NMN.

I’ve generally been feeling good with a slight increase in overall energy but nothing huge to report. But then I decided to take a further 0.5mg of MB with my last dose of NMN (about 7pm). I did some HIIT (high intensity interval training) an hour later and my performance was ridiculous. I do a pretty intense, all-out 30 secs on, 30 secs rest for 5 minutes punching and kicking the heavy bag. I’ve done this hundreds, maybe thousands of times and this was probably my best performance ever in terms of intensity and lack of fatigue. That night however my sleep was disrupted. I woke up 3 hours early (4am instead of 7am), and didn’t manage to more than doze till morning. Every time I drifted off something would jerk me awake. It felt like I might have an occasional jittery heartbeat. Sleepless nights are extremely unusual for me; in fact I don’t think insomnia like this has ever happened to me before. Surprisingly, I didn’t actually feel tired and was able to operate normally the next day (although the following night I did sleep very well indeed – without an evening dose of MB).

 

I've repeated the HIIT without taking MB (just NMN) in the evening and my performance is not as good. So I've decided to increase the morning dose of MB to 1.5mg.

 

I just heard about NMN for the first time on Rogan's podcast (#1269 with Bryan Callen). A quick google search shows it's an analogue to nicotine, I'm wondering about any possible carcinogenic effects.

Edited by marcobjj, 20 March 2019 - 11:58 PM.

[QuestforLife]

So how do we activate SIRT4 more than it would normally be activated in daily life? These are my thoughts so far:

• High NAD+
• Well-fed, no-exercise state
• Rapamycin
• L-Glutamine
• Resveratrol

 

I tried this out three times, minus the rapamycin (I decided that inhibition of protein synthesis would be counter to the aim of telomerase expression, even if it does robustly up-regulate SIRT4). 

 

I took 10-15g l-glutamine in water after a large meal and then ate a high fat yogurt with 1kg (total) of pterostilbene and resveratrol mixed in after about an hour. 

 

I also chased this with a glass of red wine before bed.

 

Two interesting results: 

 

1. The pterostilbene/resveratrol had no adverse effect on me. Normally this combination quickly makes me feel low energy with no sex drive. Quite the reverse this time! This protocol seemed to make me sleep very well that night and feel full of energy the next day.

2.  The protocol did not feel as good if I hadn't had a really nice, big meal first. Bear in mind that I'm low carb or keto, so catabolic a lot of the time. Activating SIRT4 seems to require a highly anabolic, well fed state.

 

So I'm going to continue my NMN/MB/allicin protocol in the week and then do the glutamine/resveratrol protocol at the end of the week when I have a big, carbohydrate meal.

Edited by QuestforLife, 21 March 2019 - 10:18 AM.

[Engadin]

I took 10-15g l-glutamine in water after a large meal and then ate a high fat yogurt with 1kg (total) of pterostilbene and resveratrol mixed in after about an hour. 

 

Dear Questforlife, I am afraid those are doses only whitin reach of Soros or Buffet wealthy current accounts.      Who are you?, really. 

[QuestforLife]

Dear Questforlife, I am afraid those are doses only whitin reach of Soros or Buffet wealthy current accounts.      Who are you?, really. 

 

Sorry 1 GRAM, lol!

[QuestforLife]

Before I complete my latest statin-sartan protocol and report on it, I wanted to outline my reasoning for why I want to do this. In Janic’s papers he takes patients with impaired arterial function and with 1 month of treatment with a low dose of statin and sartan, he restores that function. The benefits last at least 6 months. Based on the in vitro work on conditional programming that I have read and referred to upthread, I think it’s clear that statins are operating via ROCK inhibition to generate (in this case endothelial) progenitor cells. In another paper a similar protocol (also using mTOR inhibition) was used to de-differentiate glioblastoma cells into neural progenitors, from where they differentiated into normal neurons. So this effect is not restricted to the circulatory system, and may be body-wide, to the extent the blood stream can deliver the medication. Incidentally atorvastatin and losartan can cross the blood-brain barrier.
It is not clear in the Janic papers whether the benefits come from the presence and signalling of progenitor cells, or whether these cells carry out repairs whilst they are present (until the ROCK inhibition is removed) and this repair lasts for longer then the cells themselves. It seems unlikely to me that progenitor cells could survive for 5 months after ROCK inhibition is removed, but some residual telomerase activity is detected at 6 months, so the jury is still out.
For progenitor cells to be created, reprogramming of epigenetic marks occur to return cells to that known point (i.e. endothelial progenitor cell). This is very exciting to me, because epigenetic methylation changes are (outside of reprogramming) regarded as irreversible, because somatic cells have no way of knowing what the methylation state of a given site should be. Hence this changes over time, probably because of errors in re-methylation after DNA repair (my opinion). This may be a cause of aging. So the ability to reset that state is very interesting and there is a case for optimism that even after redifferentiation of progenitor cells, the somatic cells that result will not be returned to the previous epigenetic state – how could they be as this state is nowhere recorded – and thus will be epigenetically younger. Also, if any of the progenitor cells divide before they redifferentiate, we wouldn’t expect both daughters to reflect the epigenetic age of their original somatic ancestor. I will be submitting further myDNAge tests to determine whether I’ve affected this metric in a measurable way.
With telomere length things are not so ambiguous. We know from the conditional reprogramming papers that ROCK inhibition causes an upregulation of telomerase, and this permits further proliferation. But we also know this telomerase only maintains telomeres rather than extends them, so upon removal of ROCK inhibition and redifferentiation, cells are left with telomeres of a similar length to before treatment. This suggests that telomerase activators should be added to the protocol during the period when progenitor cells are in existence.
Even if telomeres cannot be extended, if this protocol can be repeated periodically, and Janic has shown that it can, further proliferation of cells with short telomeres can be assured.

[Andey]

Ive jumped on your bandwagon) At least on some part of it.

Today I took first 10mg atorvastatin and 12.5 losartan. I plan to take atorvastatin every other day (it have a pretty long half-life) and losartan every day.

This coincides with my goal to bring my LDLC a bit lower so I would probably continue to take low dose statin on a regular basis (or maybe titrate it up after lab results), and will discontinue losartan after two months at least for this go.

I am also doing keto/low carb, tried zero carb but it was probably too much SFA for me and my HDLC took a nose dive while LDLC stays highish anyway. So my plan for a few months while doing this intervention is to transition towards less SFA, no to zerocarb, more olive oil.

 

You wrote that you ve noticed more results doing this protocol for a month against two weeks. What are the results? ) You ve probably wrote it but I cant find it.

[QuestforLife]

I am also doing keto/low carb, tried zero carb but it was probably too much SFA for me and my HDLC took a nose dive while LDLC stays highish anyway. So my plan for a few months while doing this intervention is to transition towards less SFA, no to zerocarb, more olive oil.

You wrote that you ve noticed more results doing this protocol for a month against two weeks. What are the results? ) You ve probably wrote it but I cant find it.

I've changed my mind about keto several times and now believe it's probably not a good match for the protocol. Yes ketones are very good for the arteries, but keto makes you so catabolic any progenitor endothelial cells you create through this protocol are going to have no growth signals at all. And this matches with the second time I did the protocol - and didn't notice the same benefits I did the first time. This is all subjective of course. But I'd recommend eating well whilst doing this protocol.

I'm one day away from completing my 3rd iteration. This time I've gone for a higher dose but a shorter timescale (2 weeks). My dosing is as follows:

20mg atorvastatin AM
25mg losartan AM/PM (shorter half life)
After one-week take a single 15mg dose of Rapamycin.
Other supplements: antioxidants, some days NMN.

Report:
Felt great for a few days but then experienced a definite drop in energy and sex drive. NMN countered most of the energy loss, apart from the day after Rapamycin dosing when I was very sleepy. Also felt very chilled out, bordering on spaced out. But all negative effects seemed to wane in the second week, and now I feel really good.

I'm going to speculate that the higher dose losartan is behind the feeling of being chilled out, and my chloresterol, which was high after 4 months of keto, has probably dropped, and that might have been what affected energy and sex drive. I'm surprised Rapamycin didn't have a more obvious effect; 15mg is way higher than I've tried before.

At this point having tried several different approaches, I think I'll revert back to a 1 month protocol at the lower dose to avoid any possible side effects, and do this 2-3 x per year. And I'll probably take a high dose Rapamycin towards the end of each protocol.

Although I'm very confident I can maintain my circulatory system this way indefinitely, progenitor cells in the blood are probably not able to cross the vessel walls into the rest of the body, so I expect the benefits will only be to the blood vessels (but also indirectly will help the body with better blood supply).

Pioglitazone can create new adipocytes, which improve insulin sensitivity and glucose control, and I've tried this before, but there are some concerns with bladder cancer, so more research is warranted.

[Andey]

I have no problem gaining body weight during keto (or losing it). For me, keto just suppresses appetite a bit but it's not a problem to eat more.)

 

TBH I started to dislike simple carbohydrates and keto happens without conscious effort for me. And If I eat even 100g of carbohydrates once it doesn't break ketosis for long, probably goes to replenish glycogen first.

 

Edited by Andey, 06 April 2019 - 06:31 PM.

[QuestforLife]

I have no problem gaining body weight during keto (or losing it). For me, keto just suppresses appetite a bit but it's not a problem to eat more.)

TBH I started to dislike simple carbohydrates and keto happens without conscious effort for me. And If I eat even 100g of carbohydrates once it doesn't break ketosis for long, probably goes to replenish glycogen first.

Yes, I've gone from strict keto to low carb and it's a actually pretty hard to stay out of keto unless you REALLY hit the carbs. As for putting ON weight with keto, surely that's impossible?!

You'll probably have to eat carbs to get your body really GROWING. And that might be necessary for this protocol. I found the protocol worked well in the first month of keto but not the fourth, by which time I was depleted as hell. If you want to stay low carb I'd recommend just eating carbs in the evening and/or at weekends.

Interested to hear how you get on.

Edited by QuestforLife, 06 April 2019 - 07:02 PM.

[Andey]

Yes, I've gone from strict keto to low carb and it's a actually pretty hard to stay out of keto unless you REALLY hit the carbs. As for putting ON weight with keto, surely that's impossible?!

You'll probably have to eat carbs to get your body really GROWING. And that might be necessary for this protocol. I found the protocol worked well in the first month of keto but not the fourth, by which time I was depleted as hell. If you want to stay low carb I'd recommend just eating carbs in the evening and/or at weekends.

Interested to hear how you get on.

 

It probably depends on a diet composition (at least for me its the case)

When I was zerocarb it was hard to eat enough meat as beef is lean (no grain feeding here) and I don't like pork, so I lost some weight.

My weekly shopping list was about 5kg of beef and I added some butter or bacon to try to maintain calories. Liver was a  no go as it shutdowns appetite for a day for me while being very low calories by itself.

I am pretty lean though so my thyroid shutdowns body temperature when there is a caloric deficit for a few days in a row. And retrospectively I had some water retention going on. In a nutshell, it was a miserable experience for me even not considering that my labs went in the wrong direction.

 

When I am on my usual keto diet, I eat meat + a lot of olive oil (&dark chocolate as a dessert or a snack). I buy around 2.5 kg of beef a week(+ some liver and fish). In this configuration I struggle not to gain weight as 3 meals of this is too much considering that 100g of oil is 800kcals.

Usually, I am trying to cut down to 2 meals a day when eating this way.

 

BTW `in a wild` people take losartan 1 time a day as some metabolite thing going on. 50mg a day is an average dosage for treating HBP.

Edited by Andey, 07 April 2019 - 07:10 AM.

[QuestforLife]

That's very interesting Andey. Do you have any particular olive oil, given that you drink so much? It's a very good idea btw - I struggled to get enough fat; and mono unsaturated fat is healthy as far as I'm concerned.

I only upped losartan to twice a day as it has a short half life compared to atorvastatin; if there is a problem doing this, it's the first I've heard of it.

[Andey]

That's very interesting Andey. Do you have any particular olive oil, given that you drink so much? It's a very good idea btw - I struggled to get enough fat; and mono unsaturated fat is healthy as far as I'm concerned.

I only upped losartan to twice a day as it has a short half life compared to atorvastatin; if there is a problem doing this, it's the first I've heard of it.

TBH I buy whatever is less expensive just making sure its not an unknown brand, its an extra virgin oil from the recent harvest in a tinted glass or tin.
Fancy oils tends to be too peppery to eat a lot and costs 2x
on losartan I am saying that metabolytes stays longer than losartan itself so usually its enough to take it once a day(at least I ve read so in the drug manual)

[QuestforLife]

My dosing is as follows:

20mg atorvastatin AM
25mg losartan AM/PM (shorter half life)
After one-week take a single 15mg dose of Rapamycin.
Other supplements: antioxidants, some days NMN.

I forgot to say, for this iteration only I added MitoQ to the protocol. It has evidence for vascular renewal (see https://www.ahajourn...t=cr_pub=pubmed), but as described in the paper the effects lasted only as a long as the dosing, so it's probably only useful as an addition to the statin-sartan protocol.

[QuestforLife]

Just wanted to post a quick link to another thread someone started on skin rejuvenation. One of the chemicals identified is a ROCK inhibitor, and a quick search shows statins have been used topically for improved wound healing.

https://www.longecit...on/#entry872303

So I think it's worth having a go at creating a topical statin solution. At the moment i only have atorvastatin, which is a little heavy for skin penetration, but is lipophillic, so should dissolve well in lots of skin creams.

[Andey]

Could be of interest for you, just come out on youtube

Jerry Shay, University of Texas Southwestern Medical Center, presenting 'Telomeres and Telomerase  in Aging and Cancer ' at Undoing Aging 2019

[QuestforLife]

7 months after my last methylation DNAage test I got another one.

I deliberately took the samples just after I had completed the last statin-sartan course March, and there has been an improvement, albeit a small one.

Apparently I am now only 38 epigenetically, 2.4 years less than my actual 40 years.

My previous result, 7 months prior, said I was only 0.7 years less than my chronological age at that time (39).

So some tentative evidence for my protocol being effective.

[Andey]

7 months after my last methylation DNAage test I got another one.

I deliberately took the samples just after I had completed the last statin-sartan course March, and there has been an improvement, albeit a small one.

Apparently I am now only 38 epigenetically, 2.4 years less than my actual 40 years.

My previous result, 7 months prior, said I was only 0.7 years less than my chronological age at that time (39).

So some tentative evidence for my protocol being effective.

 

  I assume ideal epigenetic clock correlates best of all with chronological age, simply because of methodology determining correlations with ...chronological age. Also changes that small in a n=1 context scream insignificance.

 

Have you tried a Levin`s PhenoAge clock algorithm?

https://www.ncbi.nlm...les/PMC5940111/

I found a excel calculator for it 

Attached Files

•  DNAmPhenoAge_gen.xls   21.5KB   506 downloads

[QuestforLife]

I assume ideal epigenetic clock correlates best of all with chronological age, simply because of methodology determining correlations with ...chronological age. Also changes that small in a n=1 context scream insignificance.

Most of the testamonials on Longecity are n=1. In any case my theory about statins reducing epigenetic age comes from knowledge of the action of ROCK inhbitors, and the methylation age of several others on statins. But yes, evidence is very tentative at this time.

Have you tried a Levin`s PhenoAge clock algorithm?
https://www.ncbi.nlm...les/PMC5940111/
I found a excel calculator for it

Thanks, I'll check it out.