1033 replies to this topic

[ambivalent]

Thanks, for the response quest.

 

Good point, I suppose whether we run into problems downstream on TB's stem cell protocol depends on whether we do enough fusion. I suppose a metric that would seem to matter would be the summed telomere length within the stem pool, and that could be grown. So, for example, we have one stem sell with telomere length X, which could be divided into two stem cells each of length X-d, which in turn subdivide yielding 4 stem cells of length 4(X-2d) and thus net increasing pool length for all d < 3/8 X. So would you think, so long as we're increasing the total telomere length, we're not edging closer to a dramatic cliff edge? Even if the case, there would be limited storage space, one assumes, that we would eventually run into. So, if it were about maximising telomere length of a number of stem cells in a space volume V, then how would we go about doing this optimally? It would seem the best approach would to be to fuse for an extended period before "spending".  If going through a process as TB has done which would it seems be to increase stem cell supplies while intermittently emptying them with asymmetric division, the journey is a staggered one, a drunken walk. This is assuming of course, it would take many cycles of fusion to replete the pool, but through that method we end up with a lower average than when finally repleting the stem cell reserves than if we simply embark on a mission of fusion while conserving until repletion.

 

So in other words, asssuming my mental model is correct, there are three ways in which we could end up with full capacity of stem cells toying with TB's protocol - we could just divide away asymmetically with c60 burning the candle at both ends, diminsh the telomere length and then replete the stem cell pool with telomeres of a low average. Or we could do it on the hoof to rejuvenation, TB's way, fission/fusion, as is the process to retain a higher average or still, just instensely proliferate stem cells until we reason there are no more to add beyond servicing the daily attrition, this would seem to be the best approach. Then embark on spend/replete with the average telmoere length declining but always with a full supply. 

 

I should add the worst case scenario doesn't seem to be realised, as it appears long term c60 users haven't suffered adversely, as best we can know. So, one assumes there is some response to c60 stem cell activation, assuming the theory is correct, preventing rampant depletion.    

 

When mentioning, telomerase charges mitophagy, then I assume you mean the cellular function increases - but what does this mean for our individual telomerase extended mitochondria?  On the one hand the mitochondria's chance of being taken to the trash would seem to grow because mitophagy expression ramps up, but on the other, the telomere lengthening would seem to give a defective mitochondria a chance of salvation, escaping the recycling selection criteria. Also, why does telomerase increases mitophagy?     

 

On sensecent cells. I am still not seeing something of the process. Many senescent cells are it seems induced direct into senescence from other sensecent cells and senescence is triggered by telomere length and from that paper, it would seem the shortest telomere length. Apoptosis, too is characterised by short telmoere length, so why one and not the other and it seems pretty clear that senescence is a final destination, not a calling station on the way to apoptosis, not without some considerable help. Also what about extending telomeres on senescent cells, do we know what happens, would this make life tougher for senolytics? Do they behave better with telmoeres lengthened?   

 

I wonder how would be best to incoorporate telomere lenthening into TBs protocols if we had to do it. It would seem that a campaign of inducing apoptosis in near death cells as well the removal of senolytic cells would be good, then telomere lengthening, then extended fusion, followed by the stem cell protocol.  

 

Anyhow, I shall look into telomeres a little more, and at to your thread.  

 

And the questions are not all directed towards you answer, I will have a look sometime myself. 

 

 

  

[kurt9]

I wonder how would be best to incoorporate telomere lenthening into TBs protocols if we had to do it.

 His protocol does include it. You take Astragalus Root with the fusion stack every 10th round.

[QuestforLife]

Thanks, for the response quest.

Good point, I suppose whether we run into problems downstream on TB's stem cell protocol depends on whether we do enough fusion. I suppose a metric that would seem to matter would be the summed telomere length within the stem pool, and that could be grown. So, for example, we have one stem sell with telomere length X, which could be divided into two stem cells each of length X-d, which in turn subdivide yielding 4 stem cells of length 4(X-2d) and thus net increasing pool length for all d < 3/8 X. So would you think, so long as we're increasing the total telomere length, we're not edging closer to a dramatic cliff edge?

Maybe...But then again, why would you expect any protocol that increases stem cell division to increase your lifespan? The only way this could be true would be if they are not a finite resource.

To be fair we don't actually know the benefits of TB's protocol come from stem cell division, that is all hypothesis.

[QuestforLife]

His protocol does include it. You take Astragalus Root with the fusion stack every 10th round.

I know. He originally included it every round (at my suggestion) but it increased his methylation-age, same as what happened to me with epitalon and my statin-sartan protocol (d'oh). He now also uses AKG, so it may be less of an issue.

[ambivalent]

Maybe...But then again, why would you expect any protocol that increases stem cell division to increase your lifespan? The only way this could be true would be if they are not a finite resource.

To be fair we don't actually know the benefits of TB's protocol come from stem cell division, that is all hypothesis.

 

Why wouldn't we optimising a finite resource impede increased lifespan, that seems quite a reasonable proposition. If the number of stem cells impacts on lifespan, then increasing stem cells should be a candidate mechanism to improve lifespan, but of course it isn't just the numbers, as you have pointed out, but the telomere length. The value of the stem cell pool might represented to be a function of average telemore length and number of stem cells - and and the total telomere length at maximum stem cell storage capacity, might rmap to the upper bound of the potential

value achievable. 

 

Sure, this doesn't give us indefinite lifespan but certainly could extend youthspan as well lifespan. And I would guess, we might perhaps trade it off as we wish.

 

I agree there isn't enough evidence to support TB's theory, but his results won't be a fluke - even if we haven't seen repetition, that I know of. 

[QuestforLife]

Why wouldn't we optimising a finite resource impede increased lifespan, that seems quite a reasonable proposition. If the number of stem cells impacts on lifespan, then increasing stem cells should be a candidate mechanism to improve lifespan, but of course it isn't just the numbers, as you have pointed out, but the telomere length. The value of the stem cell pool might represented to be a function of average telemore length and number of stem cells - and and the total telomere length at maximum stem cell storage capacity, might rmap to the upper bound of the potential

value achievable. 

 

 

Of course, if you double the number of cells, you’ll almost double the total telomere length. But if you believe telomere lengths in stem cells shorten with divisions, then it is only a matter of how many divisions you are permitted before you die.
If we make certain assumptions, like stem cells lose 60bp/division, and they each divide once a year, and that you are born with telomere lengths of 10,000bps but die at 5,000bps [1] – all reasonable, but not unassailable, assumptions - then this gives you 83 years of life.
If TB is doing his protocol once per week, and 10% of his stem cells are dividing in response to the protocol each time, assuming he started in middle aged (7.5kbp telomere length remaining), then this protocol will kill him in 420 weeks or 8 years.
What could be wrong with this argument?
Maybe stem cells only divide every other year or only lose 30bp/division, so we have 83 years of reserve capacity… Maybe TB’s protocol only stimulates 5% of stem cells per iteration, so he can do it for longer. 
But food for thought…
[1] DOI 10.1182/blood.2021014299.

[ambivalent]

What could be wrong with this argument?

 

 

The disagreement is with the initital contention that we shouldn't expect life extension through stem cell division because stem cells are a finite resouce. 

 

Unless, I am mis-modelling, the numbers are finite but not fixed because of the potential for symmetric division. There is a space confined theoretical upper bound which we could hope to attain. I am assuming that 8, say, third generation stem cells with reduced telomere length, has greater utility than one undivided stem cell - we can make this happen. Through fusion we can be generate additional stem cells. With fasting this happens, but presumably supply meets demand, and there is little repletion.

 

And so through the increase in symmetric division generate by fusion, there comes, perhaps, the possibility to extend life/health/youth span with increased stem cell numbers and net (pool) telomere length. And that is an impovement, and we hope worth having.

 

Certainly, you're right we could burn at an alarming rate and lose them all rapidly, but that doesn't seem to be a risk with Turnbuckle's protocol unless the theory is wrong. 

 

If C60 is responsible for depleting stem cells as Turnbuckle claims, then unless the mechanism of this aspect of the process is somehow altered by the protocol, then he shouldn't have too much to worry about in 8 years given, if I recall, he only uses it at most once a week, low dose, and people have taken it daily for years. 

 

Users aren't getting the benefits of TB taking c60 daily for lengthy periods despite obviously still retaining plenty of stem cells, so it would seem likely there is some regulation - which is good. 

 

One of the questions is why not just replete without using c60 for a few months, indeed why not just fuse periodically? Are we saying that if we replete stem cells, without deploying them through c60, there would be no longevity benefit?

 

It seems quite likely there must be singalling of age to resource - "I need this much to manage a steady decline over the next 20 years", say.  How would doubling this resource, without ever using c60 to liberate it, impact health and lifespan? 

[QuestforLife]

The disagreement is with the initital contention that we shouldn't expect life extension through stem cell division because stem cells are a finite resouce. 

 

 

 

If aging is due to insufficient stem cell numbers because of too much differentiation and not enough replacement, then increasing stem cell numbers if obviously an unalloyed good.

 

If aging is due to insufficient stem cell numbers because of telomere depletion, (or some other harm associated with cell division,) then increasing stem cell numbers is obviously a bad idea. 

 

In either case you would expect a short term improvement. In the second case this would be succeeded by a rapid decline.

 

Short lived animals have faster dividing stem cells than longer lived animals. For example, in mice HSCs are reckoned to divide once per 2.5 weeks, for cats once every ~8 weeks, for baboons and macaques around every 30 weeks and 

for humans about once a year [1], [2], [3]. Of course, these figures are not 100% reliable, and future studies may adjust them somewhat, but the difference between species seems clear and is closely coupled to their lifespan. 

 

Whether this is due to telomere depletion, or other harm, at this point is not relevant. Reducing cell division is a clear path to longevity.

 

[1] Shepherd BE, Kiem HP, Lansdorp PM, Dunbar CE, Aubert G, LaRochelle A, Seggewiss R, Guttorp P, Abkowitz JL. Hematopoietic stem-cell behavior in nonhuman primates. Blood. 2007 Sep 15;110(6):1806-13. doi: 10.1182/blood-2007-02-075382. Epub 2007 May 25. PMID: 17526860; PMCID: PMC1976353.

 

[2] Catlin SN, Busque L, Gale RE, Guttorp P, Abkowitz JL. The replication rate of human hematopoietic stem cells in vivo. Blood. 2011 Apr 28;117(17):4460-6. doi: 10.1182/blood-2010-08-303537. Epub 2011 Feb 22. PMID: 21343613; PMCID: PMC3099568.

 

[3] Lansdorp PM. Telomeres, aging, and cancer: the big picture. Blood. 2022 Feb 10;139(6):813-821. doi: 10.1182/blood.2021014299. PMID: 35142846; PMCID: PMC8832478.

[ambivalent]

"If aging is due to insufficient stem cell numbers because of telomere depletion, (or some other harm associated with cell division,) then increasing stem cell numbers is obviously a bad idea."

 

This isn't clear to me, if my understanding is correct, it would seem to be a trade off. If we took a stem cell with say 10 divisions left, and in its place forced from it 8 stem cells each with 7 divisions remaining, and say those additional stem cells are "mothballed", then when our parent or "in use" stem cell is down to four divisions, we have 7 stem cells with 7 divisions quietly waiting, and the couterfactual reality would have been one stem cell with 7 divisions with no backups. If we could find a way of removing the parent cell, forcing one of the reserves into action, then the parallel scenarios are biolgically identical save for the addtional now 6 reserves with 7 divisions.

 

Eventually, as the the two parallel cells march downwards we could theoretically remove both and have in one world a 7 division replacement, while in the other a scramble for resources. Or something like that :o)

 

I suppose, I'm imagining, while we would rapidly create more older stem cells from one younger stem cells, if divisions age the cell, but those biologcially older "in stasis" stem cells might eventually become, relatively, the future's younger ones. 

 

Alternatively, some of that spare capacity might be deployed off the bat as we seem to see with TB. Anyhow, this is mindlessly speculative, on my part!

[QuestforLife]

"If aging is due to insufficient stem cell numbers because of telomere depletion, (or some other harm associated with cell division,) then increasing stem cell numbers is obviously a bad idea."

 

This isn't clear to me, if my understanding is correct, it would seem to be a trade off. 

 

I can totally see the argument that aging might not be related to running out of stem cells, but to the loss of 'flow' from those stem cells. This is probably what you are driving at. For example Steve Perry, aka Mr. GDF11, clearly thinks that stem cells need to differentiate more. For TB, he thinks they need to maintain their own numbers better (and then differentiate more). This captures what you are driving at I think - that somehow the body is not maintaining the right number of stem cells.

 

The problem is that science doesn't seem to be driven by answering the pertinent scientific questions. In the late 90s and early 00s telomeres were all the rage and many scientists were of the 1st opinion (that we ran out of stem cells). Now, fashion, not science, has decided that no, aging is probably be something else, like loss of differentiation potential, for example. But no one has done the work to eliminate either of these hypotheses. I would welcome proof either way. But no, science is now all about some nice consensus we must all arrive at through decades long dialogue.  But no discoveries are made that way. 

[ambivalent]

Well, probably my query is how to replete optimally, with fusion, assuming no telomere extension. We may want more stem cells, but we may be cautious in having more with loss of telomere length. Perhaps its worth doubling the stem cell population retaining a higher telmoere length, rather than quadruple it, say. And too if we had a target number of stem cells in mind, what would be the best approach to create a population of X stem cells while maximising telomere length. TB's approach wouldn't seem best for this aim, we presumably would prefer to create symmetric division through fusion over a period of time, and so build up a population X with a longer telmomeres,  than stagger up to X, through attrition and repletion, arriving I would imagine with a lower telomere average.    

 

The question relates to flow as you would put it, TBs approach is to increase the stem cell levels then wake then force their deployment through c60. I wonder what would happen if say our stem cell reserves tripled over night - will the taps automatically turn up, or is the flow signalled  elsewhere and remains the same? If c60 is acting in the way that TB believes it to be, then we're never seeing how the body indepdendently responds to just having those increased stem cell resources, which is not to criticise the c60 approach, but leaves us without an important observation.

 

Yes, you're right about aging theories, and I am rather glad I don't have one, as I'd likely be a sucker to it - its tougher to interrogate, be nimble, if we are so invested to it.

Edited by ambivalent, 14 October 2022 - 02:01 PM.

[Learner056]

QuestForLife,

a) I have a memory that you said something about Glutamine that it furthers (or maybe hinders, I forgot) mito Fusion, could you clarify that role in Fission vs Fusion?

b) Alpha keto gluturate is a co-factor for DNA and Histone demethylases.  Would glutamine be poor mans AKG (due to their intra-conversion) or there are too many confounders there to reasonably equate the two. 

[Learner056]

c) This might not be reported in any literature or maybe outright silly. In regards to the mitotic spindle orientation's role for symmetric vs assymetric SC division.  Is there a practical "application" of human body position - like laying down horizontal vs standing vertical, and maybe that combined with an application of compression (as that may be affecting the niche contact with SC population)?

[QuestforLife]

QuestForLife,

a) I have a memory that you said something about Glutamine that it furthers (or maybe hinders, I forgot) mito Fusion, could you clarify that role in Fission vs Fusion?

b) Alpha keto gluturate is a co-factor for DNA and Histone demethylases.  Would glutamine be poor mans AKG (due to their intra-conversion) or there are too many confounders there to reasonably equate the two. 

 

Your memory serves you well, here is the post

 

My current thinking is that AAKG supplies the arginine and the glutamine (well AKG, but that is where I think the glutamine is ending up), so you'd just need to add leucine to achieve 'hyperfusion' of mitochondria. That an avoiding the other AAs for a half a day or so. Probably explains why TB's results got much better after he added AKG. 

 

AKG is better than glutamine in my personal experience.  

[Learner056]

Thank you for that link (I can no longer juggle and provide links, it is a skill gets harder with age). I like that you don't get stuck in a 'fixed' thinking, as most people/scientists do.   

 

d) I am trying to understand the Fasting vs Fed state dynamics.  I feel that anti-oxidants (say ascorbate) intake benefit may have to do with it, is it? Like if taking ascorbate is it better to take it fasting state vs fed state.  I read some research while ago that Tylenol toxicity is very high in fasting state.  

 

 

 

add leucine to achieve 'hyperfusion' of mitochondria. 

 

[QuestforLife]

Thank you for that link (I can no longer juggle and provide links, it is a skill gets harder with age). I like that you don't get stuck in a 'fixed' thinking, as most people/scientists do.   

 

d) I am trying to understand the Fasting vs Fed state dynamics.  I feel that anti-oxidants (say ascorbate) intake benefit may have to do with it, is it? Like if taking ascorbate is it better to take it fasting state vs fed state.  I read some research while ago that Tylenol toxicity is very high in fasting state.  

 

You lose the ability to burn fats if you lack proper anti-oxidants, as seen by the benefits of the GlyNAC trial [1]

 

In our earlier studies, we discovered and reported that inducing GSH deficiency results in impaired mitochondrial fatty-acid oxidation. This suggests that
GSH adequacy is critically necessary for adequate mitochondrial fatty-acid oxidation (17). The OA in our RCT had both impaired mitochondrial fatty-acid oxidation and elevated mitochondrial glucose oxidation. This indicates mitochondrial dysfunction. The full extent of the age-associated mitochondrial impairment only became
clear when the molecular data were examined. Mitochondrial function was severely compromised on multiple levels, including ab-

normalities in mitochondrial biogenesis (PGC1α), nutrient sensors regulating fatty-acid entry into mitochondria (pAMPK/AMPK,
SirT3, CPT1b), mitochondrial β-oxidation of fatty-acids (HADHA, PPARα), electron transport chain complexes (I, II, III, V), ATP syn-
thesis (ATP5A), and mitophagy (PINK1). GlyNAC supplementation began to rapidly improve these defects in 2-weeks, but a longer dur-
ation of 16-weeks was needed to correct these defects.

 

 

Probably why C60 is so beneficial in Turnbuckle's protocol.

 

As for Tylenol, not suprising it would be more toxic to the liver when fasting, given liver will be processing more fat in that state.

 

[1] Kumar P, Liu C, Suliburk J, Hsu JW, Muthupillai R, Jahoor F, Minard CG, Taffet GE, Sekhar RV. Supplementing Glycine and N-Acetylcysteine (GlyNAC) in Older Adults Improves Glutathione Deficiency, Oxidative Stress, Mitochondrial Dysfunction, Inflammation, Physical Function, and Aging Hallmarks: A Randomized Clinical Trial. J Gerontol A Biol Sci Med Sci. 2022 Aug 17:glac135. doi: 10.1093/gerona/glac135. Epub ahead of print. PMID: 35975308.

[Learner056]

Thank you Quest.  I know I have to be wrong here, but I cannot see why, really appreciate your eyes/correction:

 

Learning1:  We hear about induced pluripotent SCs method through delivery/activation of various transcription factors: Oct4/Sox2/cMyc for rapid iPSC formation

Learning2:  We also hear that certain senolytic agents (apigenin, quercetin, luteolin etc) can play a role in "utilizing" stem cells through apoptosis.  These are usually NAD+ sources. 

 

Here is a research that shows: a) Apigenin and Luteolin viably activates Oct4, Sox2, cMyc.  b) restrains cell proliferation / increased apoptosis / arrested SCs in G2/M and S phase, c) and repressed lineage specific differentiation

 

To me, b and c are contradictory.  To me, restraining proliferation and repressing differentiation = cell cycle arrest (and certainly not SC proliferation).  To me this is nonsense. 

To me, you don't induce pluripotency by inducing apoptosis, you induce pluripotency by inducing proliferation?

 

(and off course when I say 'to me', I mean me as you and we all, as this knowledge, that I now have is through you and all here)

 

"Our results demonstrated that luteolin/apigenin restrained cell proliferation, increased apoptosis, 

and arrested PDLCs in G2/M and S phase. Luteolin and apigenin activated expression 

of Oct-4, Sox2 and c-Myc in a time and dose-dependent pattern, and repressed 

lineage-specific differentiation" https://onlinelibrar...1002/cbin.10648

Edited by Learner056, 26 October 2022 - 11:42 PM.

[QuestforLife]

 

Learning1:  We hear about induced pluripotent SCs method through delivery/activation of various transcription factors: Oct4/Sox2/cMyc for rapid iPSC formation

Learning2:  We also hear that certain senolytic agents (apigenin, quercetin, luteolin etc) can play a role in "utilizing" stem cells through apoptosis.  These are usually NAD+ sources. 

 

Here is a research that shows: a) Apigenin and Luteolin viably activates Oct4, Sox2, cMyc.  b) restrains cell proliferation / increased apoptosis / arrested SCs in G2/M and S phase, c) and repressed lineage specific differentiation

 

To me, b and c are contradictory.  To me, restraining proliferation and repressing differentiation = cell cycle arrest (and certainly not SC proliferation).  To me this is nonsense. 

To me, you don't induce pluripotency by inducing apoptosis, you induce pluripotency by inducing proliferation?

 

(and off course when I say 'to me', I mean me as you and we all, as this knowledge, that I now have is through you and all here)

 

"Our results demonstrated that luteolin/apigenin restrained cell proliferation, increased apoptosis, 

and arrested PDLCs in G2/M and S phase. Luteolin and apigenin activated expression 

of Oct-4, Sox2 and c-Myc in a time and dose-dependent pattern, and repressed 

lineage-specific differentiation" https://onlinelibrar...1002/cbin.10648

 

 

It is not understood, but senescence and/or apoptosis seems to be involved in the de-differentiation process. When inducing pluripotency via OSKM you always get some cells that become senescentt, and this appears to be important to the process, sort of like how senescent cells are involved in wound healing because they spur growth in the surroundings.

 

I did a lot of work earlier in this thread into conditional reprogramming - where using a rho kinase inhibitor and telomerase activator partially de-differentiates and immortalises (temporarily) - the cells in the culture, allowing them to proliferative indefinitely. Initially the telomerase part of the euqation was supplied by senescent mouse ESCs - so called feeder cells. It was later found that irradiating these cells produced apoptosis products that leaked into the serum, and this included telomerase [1].

 

 

 

Both feeder cells and Rho kinase inhibition are required for the conditional reprogramming and immortalization of human epithelial cells. In the present study, we demonstrated that the Rho kinase inhibitor Y-27632, significantly suppresses keratinocyte differentiation and extends life span in serum-containing medium but does not lead to immortalization in the absence of feeder cells. Using Transwell culture plates, we further demonstrated that physical contact between the feeder cells and keratinocytes is not required for inducing immortalization and, more importantly, that irradiation of the feeder cells is required for this induction. Consistent with these experiments, conditioned medium was shown to induce and maintain conditionally immortalized cells, which was accompanied by increased telomerase expression. The activity of conditioned medium directly correlated with radiation-induced apoptosis of the feeder cells. Thus, the induction of conditionally reprogrammed cells is mediated by a combination of Y-27632 and a diffusible factor (or factors) released by apoptotic feeder cells.

 

 

It may well be that compounds that induce de-differentiation induce a stage of arrest that is followed by renewed differentiation of a subset of rejuvenated cells.

 

[1]Palechor-Ceron N, Suprynowicz FA, Upadhyay G, Dakic A, Minas T, Simic V, Johnson M, Albanese C, Schlegel R, Liu X. Radiation induces diffusible feeder cell factor(s) that cooperate with ROCK inhibitor to conditionally reprogram and immortalize epithelial cells. Am J Pathol. 2013 Dec;183(6):1862-1870. doi: 10.1016/j.ajpath.2013.08.009. Epub 2013 Oct 3. PMID: 24096078; PMCID: PMC5745544.

Edited by QuestforLife, 28 October 2022 - 08:15 AM.

[QuestforLife]

Final telomerase protocol

 

I am currently running this only once a week. This is because it is fatiguing after a number of days. It is possible to run it more often, but GDF11 should only be run once per week. Epitalon at most twice a week. 

 

This protocol may be contraindicated with TB’s C60 protocol by increasing cell division of all the spawned progenitors, leading to an increase in epi age, although AKG decreases the chance of this. Nevertheless maintaining telomeres is advisable if increasing stem cell numbers. Contrary to TB’s advice, I would recommend separating telomerase activation from stem cell stimulation by atleast 2 days.   

 

I advise using AKG alongside this protocol, as all the cell division can cause glutamine deficiency. In some cases direct glutamine supplementation may be necessary. My testing seems to show that this protocol does not lead to an increase in epi age. It makes you immune as far as I can tell to viral infections, including covid. Exercise tolerance is increased. I also noticed an increase in hair strand density. This was reversed when I added an mTOR inhibitor to the protocol (to decrease proliferation and increase telomere lengthening), so that has now been removed. 

 

Protocol Details

 

GDF11 15pg subQ

Epitalon 1-2mg subQ

TAM818 1-2x 375mg softgels

Cycloastragenol 10mg capsule

Silymarin 450mg softgel

Gotu kola extract ~40% triterpenes 120mg tablet

Korean Ginseng 10:1 extract 300mg capsule

 

Rationale

 

GDF11 TERC activation (studies have shown TERC and TERT together > TERT alone >>> TERC alone); be advised GDF11 dose is highly individual 

Epitalon - antioxidant with telomerase increasing effects (latter is controversial but is possibly due to an indirect ribosomal effect on telomerase sequestering and assembly)

TAM818/Cycloastragenol/Silymarin - all activate TERT gene to various degrees by various (possibly different) mechanisms. Adding cycloastragenol certainly seemed to add to the TAM818 effect. In addition, Silymarin is liver protective, which may be required with the gotu kola. 

Gotu kola extract is highly concentrated for madecassoside and asiaticoside, the former has been shown to increase telomerase in T cells to a highly significant degree. Dosing is tricky, and more is not better, so I stick to one tablet only

Ginseng seems to have antioxidant and telomerase increasing effects, particularly in combination with cycloastragenol; also increases blood flow so is likely a ‘synergiser’

 

 

  

 

Edited by QuestforLife, 28 October 2022 - 09:17 AM.

[QuestforLife]

I have two new epi Age results from June and Oct this year. The June result is par for the course, between 5-6 years below my chronological age, as has been the case for the last 2 years. The second result from Oct this year shows a disastrous rise in biological age to 1.8 years above my biological age. This represents an acceleration in biological age of 7.3 years in 3 months! 

 

So what was the difference? I started trying Turnbuckle's Protocol 1/week. I left several days between my telomerase activation protocol and stimulating stem cells, but to no avail. 

 

I've opined before on how and why these protocols are contraindicated. I am now genuinely interested in what Longecity thinks I should do.

 

1. Cease the TA protocol while I restore my epi age. I successfully did this in 2020 with AKG, when adding epitalon to my statin+sartan protocol increased my epi age. In this case I'd probably persist with TB's protocol 1/wk.

 

2. Cease TB's protocol and continue with TA.

 

3. Forget epi age tests; they are hype and don't represent anything real? Bear in mind I've felt and looked fine during this period of accelerated epigenetic aging and my BP, HR, HRV and reaction times have shown no deterioration. My swimming times have gotten faster due to continued training.

 

4. Something else?

 

Here is a link to a Google voting form: https://forms.gle/Dp6Rcr6jXUJ71AKU7

 

Results Chart attached.

 

Attached Thumbnails

• 

[johnhemming]

Personally I am not sure about the reliability of Epi Age tests, they can be quite variable even from the same person at the same time.  I focus on biomarkers and functional tests.  To that extent the Levine algorithm, Aging.ai and various other biomarker based algorithms are I think useful.  However, in the end I am more concerned about the implications of biomarkers than any one algorithm.

 

Consider Albumin, for Levine more albumin is good, but in fact it is a U shaped curve linked to mortality.

 

 

[dlewis1453]

I feel your frustration. Do you have any recent telomere test results? Would be interesting to see if your telomeres became much longer at the same time as your epi age increase. 

 

I was thinking that splitting up TB's stem cell protocol and your protocol on a quarterly basis could be good, while continuing with AKG and GDF11 year round. So, Quarter 1 (TB), Quarter 2 (TA). Quarter 3 (TB), Quarter 4 (TA). 

[QuestforLife]

Personally I am not sure about the reliability of Epi Age tests, they can be quite variable even from the same person at the same time.  I focus on biomarkers and functional tests.  To that extent the Levine algorithm, Aging.ai and various other biomarker based algorithms are I think useful.  However, in the end I am more concerned about the implications of biomarkers than any one algorithm.

 

Consider Albumin, for Levine more albumin is good, but in fact it is a U shaped curve linked to mortality.

 

I've taken enough tests with this company to understand what their error bars are, and this recent test is a real result.

 

What remains is to determine what the practical significance is. 

 

My working hypothesis is that I am generating sufficient telomerase such that the downstream cells used in this test are dividing significantly more times than is usual, and this is having the effect (in the test) of mimicking an older person - who replaces their somatic cells from their stem cell reserve less often than a younger person. If this is true then the practical significance of this result is limited, I merely need to wait until these cells are cleared and my epi age will return to normal.  This is pretty much what happened last time in 2020 with epitalon. 

 

The reason that this effect is only seen when stimulating stem cell division, I believe, must be due to the significantly greater number of progenitor cells in circulation. Ordinarily a telomerase activator does not have a significant effect on epi age, but only when many more cells capable of significant division are created. You are essentially creating a lump of cells that move through the body in greater number than would ordinarily be the case. My understanding of epigenetic aging is that it affects all cells, dividing and non-dividing, but the non-dividing part is being completely missed by these tests, and/or it is all down to division history. A more nuanced answer might be that as cells differentiate they get an 'older' epigenetic age - this is probably some fixed value for fully differentiated, non-dividing cells - maybe drifting up slowly as a function of metabolic health decline, but that the bulk of epigenetic aging is actually created by dividing cells as they get further from the stem cell blueprint. It is an idea anyway. 

[QuestforLife]

I feel your frustration. Do you have any recent telomere test results? Would be interesting to see if your telomeres became much longer at the same time as your epi age increase. 

 

I was thinking that splitting up TB's stem cell protocol and your protocol on a quarterly basis could be good, while continuing with AKG and GDF11 year round. So, Quarter 1 (TB), Quarter 2 (TA). Quarter 3 (TB), Quarter 4 (TA). 

 

Both Turnbuckle and I have the aim of helping stem cells. 

 

The problem is each only divides every year or so (in humans). So I aim to use telomerase activators a lot - and therefore catch dividing stem cells as often as possible with extra telomerase. But the downside is that most cell division is by the direct descendants of stem cells, and these get most of benefit of the telomerase present. So I am feeding a bunch of hungry hatchlings, whilst trying to get a little bit of food to the mother hen. 

 

TB is trusting that stem cells can take care of their own telomerase (his occasional addition of a little astralagus extract is of no utility), but leaving all the downstream cells to divide the normal amount. So he is working the mother hen harder, and avoiding feedings all those rapacious chicks. (You could also argue that most of the symmetrical division isn't pure stem cells but progenitors - so chicks making more of themselves but not differentiating as they do it, as with the TA protocol).

 

So both approaches have problems. 

 

In answer to your question, no - Lifelength still aren't accepting blood samples from the UK (ex-EU) as the extra border checks mean they can't get them in the time required. So I am reduced to looking at proxies like Neutrophil to Leukocyte ratio, etc. 

[QuestforLife]

I was thinking that splitting up TB's stem cell protocol and your protocol on a quarterly basis could be good, while continuing with AKG and GDF11 year round. So, Quarter 1 (TB), Quarter 2 (TA). Quarter 3 (TB), Quarter 4 (TA). 

 

When I've figured out a way to go with this, you'll be the first to know!

[dlewis1453]

 

 

So I am feeding a bunch of hungry hatchlings, whilst trying to get a little bit of food to the mother hen. 

 

This is a great analogy. We just need to find some food that only the mother hen can eat. 

 

Thanks for explaining your thoughts on why you have paired the protocols in this way, and why your protocols led to this spike in epigenetic age. Your reasoning makes sense. I think for me personally, I would want to take a break from the TA protocol and continue with epi-age reducing interventions for a few months to confirm that the epi-age declines as expected. With that theory confirmed, I would then feel more confident in proceeding aggressively with the TA protocol in the future.

[QuestforLife]

Adding telomerase to symmetric division: a simplified model

 

In the attached drawing you can see that when symmetric division is stimulated, resident stem cells (small, bright blue) engage in self-renewal with most (¾) cells responding but 1 cell still dividing asymmetrically.

 

Differentiation downregulates telomerase activity in most somatic cells (large, dark blue), limiting their future replication ability. But adding extra telomerase restores replication ability to these cells, who are then able to respond to the stimulation by dividing, as seen in the lower image.

 

In this simplified example, the initial ratio of stem to non-stem cells is 1:1. After symmetrical division of (¾) cells with active telomerase the ratio of stem to non-cell is improved to 5:4. But because adding telomerase gives non-stem cells increased division capability, stimulation in this case results in the ratio decreasing to 5:7.

 

This model can explain the effect of telomerase activation when combined with stem cell stimulation, resulting in an increase in epigenetic age, which is proposed to measure the differentiation state of a cell, with more differentiated state resulting in an older measure. In a mixed tissue, the ratio of stem to non-stem cells therefore sets epigenetic age.

 

Future stem cell self-renewal stimulation should include a way of increasing telomerase in stem cells only, possible by increasing TERC. Unfortunately as most epigenetic aging tests include leukocytes, which have active telomerase, this cell type may also benefit.

 

 

Attached Thumbnails

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[QuestforLife]

Adding telomerase to symmetric division: a simplified model

 

In the attached drawing you can see that when symmetric division is stimulated, resident stem cells (small, bright blue) engage in self-renewal with most (¾) cells responding but 1 cell still dividing asymmetrically.

 

 

I’ve been thinking about this model, and ultimately I’ve had to reject it. The reason is that it doesn’t explain the importance of symmetric division in the acceleration of epigenetic age. In both circumstances when I have experienced this (statin-sartan protocol and TB protocol) the commonality was the stimulation of the division of primitive cells. The telomerase activators that are supposedly responsible for this acceleration caused no such harm on their own, i.e. during the normal differentiation process, only when I used them in conjunction with this process. The key difference seems to be symmetric division. Fatty acids stimulate that in TB’s protocol, and the inhibition of the cytoskeleton causes this in the statin-sartan protocol.

 

What do we know about symmetric division? During asymmetric division, the actin spindles separate out the chromatids preferentially, i.e. they somehow know which half of the genetic material is destined for the differentiating cell, and which is to remain as the stem cells: this is likely mediated by methylation on the surface of the genes - when the new material is copied it will naturally lack this and so this is an easy way to tell the two halves apart. If we interfere with this division process somehow, by stopping the ability to separate out the two halves of the cell in an asymmetric manner, then the cell will have no control over which half of the genetic material goes where.  This becomes random and we would expect each daughter cell to have a partially differentiated state, rather than one remaining undifferentiated and the other fully differentiated. This might make very little difference if it occurs only once. But with telomerase activators making more division possible, and with a partially differentiated state being highly permissive for division, then we can understand how symmetric division can cause epigenetic aging. There has to be more than this to it, as when stem cells divide symmetrically in normal circumstances, then you wouldn’t expect epigenetic age to accelerate - but we do know that epigenetic age is highly accelerated during development (childhood), i.e. when we are getting bigger, cells divide symmetrically in large numbers. You might ask, how then do these protocols result (when not using TAs), in a younger epigenetic age? My guess is that when the biohacker is already old, even partially differentiated cells are going to be a net improvement on what is present in the blood from day to day - so long as division in this state does not become excessive (as with the addition of TAs).

[Castiel]

iirc Sinclair said that massive induction of DNA breaks without causing mutations leads to accelerated epigenetic aging in models and consequent display of aging phenotype.

 

Recent research has shown that dependent on rate of aging of a species is rate of mutation accumulation, and that mutation accumulation can be significant iirc about 1k mutations over lifetime per cell.

https://www.genengne...light-on-aging/

This natural mutation accumulation should be accompanied by epigenetic aging just like the double strand breaks in Sinclair's experiments and this should result in consequent display of aging phenotype.

 

We know that one of the main ways in which long lived species reduce mutation accumulation is via modification of membranes to make them more oxidation resistant.

https://pubmed.ncbi....h.gov/22708840/

 

It is believed this membrane composition difference may account for the difference in longevity between birds and other animals despite birds being high metabolism animals.

 

Another thing that was recently found, don't know if I previously mentioned it, was that some species of long lived bats experience notable epigenetic rejuvenation each hibernation cycle and this may be in part behind their great longevity.

https://www.scienced...20809194910.htm

Given these bats live like 10x the expected lifespan for a rodent their size, epigenetic rejuvenation seems promising approach.

 

While on the talk of membrane composition membrane composition changes adversely through aging making it more vulnerable, iirc, whilst CR alters membranes making them more resistant to oxidation damage

 

 

 A lowered unsaturation/saturation index is observed for mitochondrial membrane lipids in calorie-restricted rodents resulting in an altered membrane structure and function. Plasma concentrations of insulin and triiodothyronine are significantly lower under calorie-restricted feeding conditions and these hormones exert transcriptional control over desaturase enzymes that are important in the control of membrane lipid unsaturation. A loss of double bonds should make the mitochondrial membranes more resistant to peroxidation damage and would also reduce the proton conductance of the membrane, raising the membrane potential at a given respiration rate. This effect however, appears to be offset by other membrane changes that may include increased activity of uncoupling proteins. These unidentified adaptations increase the proton leak in calorie-restricted animals resulting in a lowering of the membrane potential and ROS generation. https://pubmed.ncbi....h.gov/12200030/

 

 

Membrane alteration as a basis of aging and the protective effects of calorie restriction https://www.scienced...047637405000886

 

There are diverse causes of cellular senescence, but the final kingpin in the matter is of course telomeres...

yes, but like the resveratrol experiment showed, if a cell has notable genetic damage lengthening telomere will not make it exit senescence.   Only cells with little genetic damage will exit senescence upon telomere restoration.

https://bmcmolcellbi...2860-017-0147-7

That is the paper not sure if mentioned in the paper but the researcher did mention that in a youtube video.

 

I believe senolytics may be necessary for rejuvenation eventually to remove senescent cells with severe dna damage.

 

I still think protecting the membranes sounds like a promising approach assuming you don't negatively affect cell signaling or function.   Astaxanthin the most powerful membrane protector sounds promising as such, and as I mentioned earlier early preliminary data from interventions testing suggest significant lifespan extension may be observed on astaxanthin supplemented rodents.   I'd also like to see whether tocotrienols have longevity benefits or not given they too are strong membrane protectors.

 

Telomere lengthening and epigenetic rejuvenation do sound like promising approaches to extend lifespan.  But we definitely should be reducing mutation accumulation as whilst the downstream epigenetic effects are reversible, the mutations are not so far.

 

 

[QuestforLife]

 

....

 

I still think protecting the membranes sounds like a promising approach assuming you don't negatively affect cell signaling or function.   Astaxanthin the most powerful membrane protector sounds promising as such, and as I mentioned earlier early preliminary data from interventions testing suggest significant lifespan extension may be observed on astaxanthin supplemented rodents.   I'd also like to see whether tocotrienols have longevity benefits or not given they too are strong membrane protectors.

 

Telomere lengthening and epigenetic rejuvenation do sound like promising approaches to extend lifespan.  But we definitely should be reducing mutation accumulation as whilst the downstream epigenetic effects are reversible, the mutations are not so far.

 

 

There are many things going on. Astaxanthin may inhibit membrane oxidation and therefore DNA damage, but it is also an mTOR inhibitor, according to my self-experiments, so we can't directly state its benefits come from protection from membrane oxidation. That said, anything that arrests the cell, whilst it still receives growth signalling, is going to cause aging via hypertrophy of cells (they grow even when they can't divide), therefore astaxanthin may be protective from multiple angles and I would expect it to extend life in animals. 

 

Yes of course telomeres aren't going to fix everything - there is rejuventation and there is aging - everyone conflates the two but they are not the same thing. Telomeres are indispensable to rejuvenation.