2503 replies to this topic

[kurt9]

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

 

To recap. 

 

Test date...Epigenetic age - chronological age

2/2018 ... +.5 years (This was baseline, before treatments)

6/2018 ... -11

 

I began to use telomerase stimulants at this point, and my epigenetic age began to increase. When I stopped telomerase stimulants, epigenetic age stabilized and slowly began to fall with continued SC treatments. Cycloastragenol and astragalus extract screwed up my age regression for a year.

 

12/2019 ... -13

2/2020 ... -14.2

 

This is kick ass! Its not something I would do at my current stage in life. But it is nice and reassuring that something like this is available for when I need to do it some time down the road.

 

BTW, I finished week 6 of the mitochondrial fission/fusion thing. I then had to stop because I was travelling last week and am getting over a bug I picked up on the flight (No, It not COVID-19). So, I resume next week and will continue for another 3 weeks and possibly 6 weeks.

[ryukenden]

Have you reversed your grey hair with the epigenetic age reduction?

Is there alternative to stearic acid? I am worried about whether it can cause atherosclerosis.

[thenewwildone]

My latest epigenetic age test result, ten weeks after the last one, is 14.2 years below chronological age. Over that period I used the modified SC treatment on this post, with one treatment per week. The test was TruAge from Trumelabs.

To recap.

Test date...Epigenetic age - chronological age
2/2018 ... +.5 years (This was baseline, before treatments)
6/2018 ... -11

I began to use telomerase stimulants at this point, and my epigenetic age began to increase. When I stopped telomerase stimulants, epigenetic age stabilized and slowly began to fall with continued SC treatments. Cycloastragenol and astragalus extract screwed up my age regression for a year.

12/2019 ... -13
2/2020 ... -14.2

I’ve been following this topic from the beginning and I really want to believe that you’re really getting younger. The problems are that it’s possible that you’re actually depleting your stem cell pool. People who have diseases that mimic aging have epigenetic ages that correlate to their age, but their telomeres are depleting almost twice as much as normal. This tells me that telomeres maybe more important than epigenetic age.

Ultimately there are only 2 ways to prove if any method is working.

1. Actually look younger after doing a protocol/protocols.

2. Live significantly longer than the average lifespan and be able to do things that people your age can’t. This one maybe objective.

All these test results mean preciously little unless they lead to one or both of the above and for #2 we won’t know for a while. I don’t know your age, but if you’re not capable of easily doing things that people 13 years younger can do physically (can be objective) and don’t look 13 years younger, you’re not younger.

[Turnbuckle]

Have you reversed your grey hair with the epigenetic age reduction?

 

 

 

So far graying may have been arrested, but not dramatically reversed. My feeling is that, if a stem cell niche is completely depleted, it's much harder to recover.

 

 

I’ve been following this topic from the beginning and I really want to believe that you’re really getting younger. The problems are that it’s possible that you’re actually depleting your stem cell pool. People who have diseases that mimic aging have epigenetic ages that correlate to their age, but their telomeres are depleting almost twice as much as normal. This tells me that telomeres maybe more important than epigenetic age.

Ultimately there are only 2 ways to prove if any method is working.

1. Actually look younger after doing a protocol/protocols.

2. Live significantly longer than the average lifespan and be able to do things that people your age can’t. This one maybe objective.

All these test results mean preciously little unless they lead to one or both of the above and for #2 we won’t know for a while. I don’t know your age, but if you’re not capable of easily doing things that people 13 years younger can do physically (can be objective) and don’t look 13 years younger, you’re not younger.

 

The epigenetic results closely parallel physical results. See post #201. In addition to those things noted in that post, I experienced an elimination of a distortion in the Amsler grid that I'd seen for some years. The increase in shoe size I noted there finally stopped. 

[Turnbuckle]

This is an update from post #1149. The only change is with resveratrol. After I added that, I felt there was an acceleration in age reduction, but the drop I saw in the epigenetic test results wasn't significant enough to say it actually improved anything. So best to take it out for now.

 

 

Caveats:

1. This is a work in progress.

2. It is intended as a geriatric treatment for age reversal.

3. One should avoid alcohol during this treatment.

4. A link to the latest protocol can always be found on my profile page.

  

 

Part 1: Stem cell self-renewal

 

 

Time 0 —

¹Stearic acid — 5-10 g (food grade, in brownie)

 

  

Time 3:00 —

²Sulforaphane — 100 mg

³Liposomal glutathione — 1 g

⁴SAM-e — 500 mg

 

 

Time 3:30 —

⁶C60 — 3 mg (in oil)

⁷Amino acids: Threonine — 2-3 g, Lysine — 2 g, Methionine — 1 g, Leucine — 1 g

 

These amino acids should be repeated as necessary every couple of hours, and perhaps days.

 

 

Part 2: Senescent cell replacement (24 hours later)

 

⁸Nicotinamide — 2g, Ribose — 2g

⁹Curcumin (liposomal or phytosomal) — 2g

⁷Lysine — 2 g, Methionine — 1 g, Leucine — 1 g

 

These amino acids should be repeated as necessary every couple of hours, and the entirety of Part 2 may be repeated the next day as well.

 

 

 

Notes:

 

(1) This is for mito fusion, which directs SCs to self-renewal. Food grade stearic acid is a waxy triglyceride with about 50% stearic acid moieties. It has a high melting point and will have very poor availability unless properly prepared. Baking it into brownies is one option: Using a box mix that calls for 1/2 to 2/3 cups of oil, eliminate the oil and add 120 grams of stearic acid flakes or granules, leaving the rest of the recipe unchanged. Mix at room temp using a power mixer, bake according to directions on the box, then divide 3x4 and freeze most of it for later use. Use ½ to 1 brownie for this protocol. With the addition of sulforaphane, I've been using ½. Stearic acid has much longer half life than sulforaphane (11 vs 4 hours), and thus may interfere with subsequent senescent cell replacement, which requires mito fission. It may be possible to use sulforaphane alone, but I've been content with just cutting it back.

 

(2) Sulforaphane penetrates the BBB to produce mito fusion there, while stearic acid is blocked. 50 mg caps can be obtained from Amazon — Thorne Research - Crucera-SGS - Broccoli Seed Extract for Antioxidant Support - Sulforaphane Glucosinolate (SGS)

 

(3) Primary antioxidant.

 

(4) Methyl group donor.

 

(5) --

 

(6) UCP2 blocker. A teaspoon of commercially available C60 oil.

 

(7) These can be used in caps or tablets, though I use a commercial blend of essential amino acids here, adding extra lysine, methionine and leucine. Pluripotent cells have a special need for these three. I would spread out the doses as indicated, to avoid nausea. Note that I deleted threonine in the last update, but I got negative results by leaving it off. This became increasingly apparent after a few weeks, thus I restored it.

 

(8) Used together, nicotinamide and ribose rapidly increase NAD+, which drives mitochondrial fission, required for senescent cell apoptosis and replacement.

 

(9) Senolytic

Edited by Turnbuckle, 04 March 2020 - 10:10 AM.

[QuestforLife]

So far graying may have been arrested, but not dramatically reversed. My feeling is that, if a stem cell niche is completely depleted, it's much harder to recover.

 

 

 

The epigenetic results closely parallel physical results. See post #201. In addition to those things noted in that post, I experienced an elimination of a distortion in the Amsler grid that I'd seen for some years. The increase in shoe size I noted there finally stopped. 

 

Even with younger looks and performance, there is at this point, no way outside of giving a bone marrow telomere sample to determine if the bone marrow is being used up at an increased rate (as the price for greater health), or whether genuine rejuvenation has taken place.

 

On the bright side, mouse studies such as this one [https://onlinelibrar...1111/acel.13110 , see Figure 1], where there was a control group who had increased bone marrow mobilisation (but no exogenous replacement), experienced a normal lifespan (you could argue it was slightly reduced but it wasn't statistically significant). 

 

So Turnbuckle's advice that this is a geriatric treatment only, is probably wise. 

[Turnbuckle]

Even with younger looks and performance, there is at this point, no way outside of giving a bone marrow telomere sample to determine if the bone marrow is being used up at an increased rate (as the price for greater health), or whether genuine rejuvenation has taken place.

 

 

 

 

I very much doubt that telomere length of bone marrow is predictive of anything, particularly aging or longevity.

 

 

Background and aim of the work: Bone marrow failure syndromes (BMFS) includes inherited and acquired conditions.... We have found that there is a significant difference in relative telomere length between congenital group and controls (p=0.001), also a significant difference between acquired group and controls (p= 0.029). However, there is no significant difference between congenital and acquired groups (p= 0.479). There is no significant correlation between the telomere length and the overall survival or prognosis of the patients of BMFS.

https://ashpublicati...-of-Bone-Marrow

 

[QuestforLife]

I very much doubt that telomere length of bone marrow is predictive of anything, particularly aging or longevity.

 

 

 

There are diverse conditions that lead to BM failure; some of which result in shorter telomeres in circulating cells and some of which do not. See: https://www.ncbi.nlm...les/PMC4281312/

 

Most dyskeratosis congenita telomere lengths were below the 1st percentile, while only 2 Fanconi anemia and one each Diamond-Blackfan anemia and Shwachman-Diamond syndrome were that low. However, Fanconi anemia, Diamond-Blackfan anemia and Shwachman-Diamond syndrome clustered in the bottom half of the normal range

 

Moreover in the study you reference you are cherry picking the telomere lengths of an acquired vs. inherited condition, which were not significantly different from one another as you might expect, rather than the differences between either condition and unaffected controls, which was significant.

 

Finally, the abstract of your referenced study does not specify what tissue was used to measure telomere lengths. The whole crux of my argument is that you need to measure BM stem cell telomere lengths NOT a one off measurement of a dynamic population like leukocytes (as is common), the latter are influenced by a multiplicity of contradictory factors.   

[Turnbuckle]

An experiment with cultivated stromal cells found that, with multipotential stromal cells, methylation changes but not telomeric length correlate with survival--

 

Results. Regardless of donor age, cultures’ osteoprogenitor content correlated better with remaining lifespan (population doublings before senescence, PD-BS) than proliferative history (accrued PDs). Individual gene’s expression or telomere length did not predict PD-BS but methylation of individual CpG islands did...

 

In spite of literature supporting the loss of telomere length being a feature of MSC replicative ageing [9, 10, 16] no correlation was found in either culture expanded MSCs or osteodifferentiated MSCs, although the differentiated MSCs did have significantly shorter telomeres. Modelling calculations in conjunction with SPARC were attempted but were not predictive.

 

The methylation of CpG islands [12] collectively had very strong associations with PD, which remained for PD-BS albeit reduced...

https://www.hindawi....i/2017/6129596/

 

 

 

Telomere length is simply not predictive. It's only a sell-by for individual cells.

[QuestforLife]

Telomere length is simply not predictive. It's only a sell-by for individual cells.

Yes it's the rate of shortening that matters but that is besides the point. Adult stem cells are notoriously sensitive to culture conditions and in general behave quite differently in vitro to how they do in the body. You can't draw any firm conclusions from a Petri dish study.

It is well known stromal stem cells produce less osteocytes and more adipocytes with age, so that would be a good predictor of their function. The fact that this correlates with their potential population doublings in vitro indicates to me that it is related to telomere shortening. It is easy to see how this relationship might be obscured by the variation in telomere length between a small number of donors. There is also a great heterogeneity in stromal cells even within a single individual. Senescence independent of telomere length is also a factor, but this is much more common in high O2 (high ROS) culture conditions than in the body, again highlighting the caution required when interpreting in vitro studies.

[Turnbuckle]

You can't draw any firm conclusions from a Petri dish study.

 

 

And all you get from in vivo are scatter charts with only a vague trend.

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[kurt9]

Can this C60 protocol be used to repair the lung fibrosis damage that some COVID-19 survivers are reporting?

[yz69]

interesting paper "Hyperactivation of sympathetic nerves drives depletion of melanocyte stem cells"

 

https://sci-hub.se/1...1586-020-1935-3

 

 

Under conditions of stress, the activation of these sympathetic nerves leads to burst release of the neurotransmitter noradrenaline (also known as norepinephrine). This causes quiescent melanocyte stem cells to proliferate rapidly, and is followed by their differentiation, migration and permanent depletion from the niche. Transient suppression of the proliferation of melanocyte stem cells prevents stress-induced hair greying.

 

Does this mean one has to have sex under mito fusion mode so the stem cells go to self renewal instead of  differentiation?

 

 

[Graviton]

interesting paper "Hyperactivation of sympathetic nerves drives depletion of melanocyte stem cells"

 

https://sci-hub.se/1...1586-020-1935-3

 

 

Does this mean one has to have sex under mito fusion mode so the stem cells go to self renewal instead of  differentiation?

In addition, exercise is known to induce mitochondrial fission and it seems to temporarily secretes NE.

[ryukenden]

Would you consider taking the following stem cell supplement from LE with your regime? It may be helpful for you?

http://www.lifeexten...aspx?item=02401

[yz69]

Interestingly, this article suggest NE is also pro mito fission:

https://www.nature.c...es/ncb2950.pdf 

 

Norepinephrine promoted protein kinase A (PKA)-mediated phosphorylation of the pro-fission protein Drp1, and its localization with the mitochondria.

 

Any idea about how to reverse the hair greying?

 

[Biotochandron]

Must Mito dysfunction first be reversed before starting to work with the stem cell protocol? Or it is okay to do both?

 

 

[Turnbuckle]

Interestingly, this article suggest NE is also pro mito fission:

https://www.nature.c...es/ncb2950.pdf 

 

Any idea about how to reverse the hair greying?

 

It seems possible to stop the process and reduce it to some degree, but insofar as viable melanocyte SCs are completely absent, it may be impossible.

Must Mito dysfunction first be reversed before starting to work with the stem cell protocol? Or it is okay to do both?

 

 

They could be done together, but if you have significant mito damage, I suggest you not muddy the waters and attend to that first.

[ryukenden]

Interestingly, this article suggest NE is also pro mito fission:
https://www.nature.c...es/ncb2950.pdf

Any idea about how to reverse the hair greying?

I saw a news article 2-3 years ago about reversing grey hair by immune drugs unintentionally.

https://www.google.c...s-gray-hair.amp

[kurt9]

This is the reason why this protocol may be effective at dealing with COVID-19:

 

https://www.aging-us...cle/103001/text

[Turnbuckle]

This is the reason why this protocol may be effective at dealing with COVID-19:

 

https://www.aging-us...cle/103001/text

 

 

As RNA viruses like covid fill up cells and then spread their daughter virions via apoptosis, senolytic drugs may well make the disease worse. Fusion, however, may slow it down and give the body time to build an immune response. See https://www.longecit...-stop-covid-19/

[ambivalent]

If treating with senolytics appears to work why oppose with theory? Anyhow, wouldn't eradicating quantities of cytokine emitting senescent cells help reduce the category of the c-storm? 

[kurt9]

As RNA viruses like covid fill up cells and then spread their daughter virions via apoptosis, senolytic drugs may well make the disease worse. Fusion, however, may slow it down and give the body time to build an immune response. See https://www.longecit...-stop-covid-19/

 

I have personal experience with this. I came back from business travel, felt a little run down, and proceeded to do another round of the mitochondrial protocol. I ended up making myself sick as a result. This was in January. I returned from another trip in February, and skipped a week precisely to avoid getting sick a second time. So, I think you're correct on this. I think where senolytics are useful is once you recover, it you end up with lung fibrosis or other fibrosis, then you use the senolytics to fix that.

[Turnbuckle]

If treating with senolytics appears to work why oppose with theory? Anyhow, wouldn't eradicating quantities of cytokine emitting senescent cells help reduce the category of the c-storm? 

 

 

The article kurt linked to was speculative.

Edited by Turnbuckle, 13 April 2020 - 07:29 PM.

[Unclebob]

Hi guys

 

been following the Manipulating mitochondrial dynamics protocol since the start of the year (Fission followed by Fusion) and was looking at Senolytics particularly the results and protocol of the Mayo Study using Fisetin.

 

Seeing that Senolytics plays a part in the Stem Cell self renewal with C60 I decided to start this protocol with a view to following it up in a month much like the work done at the Mayo clinic. 

 

So I used Fisetin and Quercetin instead of Curcumin for the Senolytic effects and instead of Stearic Acid, Glycerol Monostearate as it is far easier to obtain in the uk.

 

Day 1 and 2

Glycerol Monostearate, Sulforaphane and C60 in EVOO

 

Day 3 and 4

NR followed by 2g of Fisetin and 1g of Quercetin

 

I am ketogentic and have been intermittent fasting for 18mths. 

 

Results:

1. Can report no knee or foot pains but felt "off" or "out of sorts" on days 3,4,5.
2. Of note is that my sports/heart/sleep/exercise smart watch seems to have burnt the skin under its laser light (found on day 5) not sore just red and a slight wee-page.  Moving the watch to the other wrist for 24hrs showed no similar effect.  Could it be a co-incidence or a reaction to the protocol?

 

Will return to the Manipulating mitochondrial dynamics protocol for the next few weeks and repeat the Stem Cell protocol towards the end of May.

 

Thank you for all the information and discussion.

 

Cheers

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Edited by Unclebob, 30 April 2020 - 09:31 AM.

[hsibai]

A recent study published in Aging Research Reviews titled “Targeting senescent cells to attenuate cardiovascular disease progression”
The study used Dasatinib + Quercitin. Would it make sense to incorporate Quercitin in the senolytic part of this protocol?
https://doi.org/10.1...arr.2020.101072

[Turnbuckle]

A recent study published in Aging Research Reviews titled “Targeting senescent cells to attenuate cardiovascular disease progression”
The study used Dasatinib + Quercitin. Would it make sense to incorporate Quercitin in the senolytic part of this protocol?
https://doi.org/10.1...arr.2020.101072

 

I've found that mito fission + curcumin + a telomerase blocker such as cinnamon extract (which contains cinnamaldehyde and eugenol) works best. This can be used with stem cell nutrients such as méthionine and lysine. I don't have experience with Dasatinib, but I'm leery of such strong drugs. I believe that removing senescent cells at a moderate rate is best, allowing the associated paracrine signaling to simultaneously stimulate replacement. I suggest shortening telomeres of adult stem cells (and TACs) as a way of replacing them faster, as they don't have zero méthylation and thus they do age. While this is speculative at the moment, it appears to be a good approach from subjective measures, and I have an epigenetic test in the works to validate it. I've already found that going the other way by lengthening telomeres accelerates epigenetic aging.  

 

This approach requires stimulating VSELs first. It is then best to do the above in the next day or two, as it is not clear how long an expanded population of VSELs will last. I believe that these circulating pluripotent cells are the basis for good results seen from parabiosis. Sometimes, anyway, as this often fails when VSELs are filtered out, or are depleted in the donor.

 

Very small embryonic-like stem cells (VSELs) represent a population of extremely small nonhematopoietic pluripotent cells that are negative for lineage markers and express Sca-1 in mice and CD133 in humans. Their embryonic-like characteristics include the expression of markers of pluripotency; the ability to give rise to cellular derivatives of all three germ-layers; and the ability to form embryoid-like bodies. Indeed, quiescent VSELs may represent the remnants of epiblast-derived cells in adult organs. After tissue injury, including acute myocardial infarction (MI), bone marrow–derived VSELs are mobilized into the peripheral blood and home to the damaged organ. Given the ability of VSELs to differentiate into cardiomyocytes and endothelial cells, and their ability to secrete various cardioprotective growth factors/cytokines, VSELs may serve as an ideal cellular source for cardiac repair.

https://www.ncbi.nlm...les/PMC3159118/

 

 

The progression goes like this: VSELs >> SCs >> TACs >> Somatic cells

If VSELs are depleted or if the telomeres of subsequent cells in the progression are extended, epigenetic age will increase.

Edited by Turnbuckle, 10 May 2020 - 01:05 PM.

[Biotochandron]

 

 

They could be done together, but if you have significant mito damage, I suggest you not muddy the waters and attend to that first.

 

Hey TB, thanks for answering to my post. Because of my condition (mito damage and collagen injuries / pain all over my body due to fluoroquinolone poisoning) I decided to add the stemcell protocol in addition to your fission protocol. And I think it's pretty much improving my condition. Is there any problem to take stearic acid and C60 more often, like 4 days straight or is there a maximum per week/month you would suggest doing stearic acid/C60?

[Turnbuckle]

Hey TB, thanks for answering to my post. Because of my condition (mito damage and collagen injuries / pain all over my body due to fluoroquinolone poisoning) I decided to add the stemcell protocol in addition to your fission protocol. And I think it's pretty much improving my condition. Is there any problem to take stearic acid and C60 more often, like 4 days straight or is there a maximum per week/month you would suggest doing stearic acid/C60?

 

Stem cells are only useful if they have something that needs replacing, and if you overfill a niche, expect that the extras will be eliminated by homeostatic mechanisms and thus wasted. So I suggest once a week.

[Andey]

 

Interesting info here - liver sinusoid endothelial cells (LSEC) gets senescent early(or dont get removed efficiently) and during a study with senolitic treatment they get replaced by fibrotic tissue, this affecting the liver function.

Another problem - with massive senescent cells removal capillary system loses integrity and leads to blood leaking.

 

One could argue that we dont really want too effective senolitic treatment unless this is resolved. Then we probably want something that supresses fibrosis (I believe curcumin shown to decrease liver fibrosis), we need to be sure that we have enough stem cells in a pool and probably need some help with differentiating into correct cell types.

Maybe addition of phospatidylserine would help? I see a small bunch of studies where it helps to differentiate into correct cell types for different environments.