2503 replies to this topic

[QuestforLife]

 

Keeping in mind this study was done with mice, it appears there are two ways to improve pluripotency: with supplemental glutamine, or with α–ketoglutarate (and presumably, various precursors thereof). In adults, this should improve the function of VSELs -- the pluripotent backup to adult stem cells and the main target for the protocols of this thread.

 

Given that valine, threonine, methionine and isoleucine feed into the krebs cycle at succinyl-coA, the step between AKG and Succinate, we might now want to avoid those amino acids if we really believe the succinate:AKG ratio is what is important in maintaining pluirpotency. Or might it be that such AAs relieve fatigue during the protocol precisely because they boost succinate? 

 

Leucine however increases the conversion of glutamate to AKG, so might be desirable. 

 

https://link.springe...0726-016-2254-8

 

 With the help of transport systems, extracellular L-glutamine crosses the plasma membrane and is converted into alpha-ketoglutarate (AKG) through two pathways, namely, the glutaminase (GLS) I and II pathway. Reversely, AKG can be converted into glutamine by glutamate dehydrogenase (GDH) and glutamine synthetase (GS), or be converted into CO2 via the tricarboxylic acid (TCA) cycle and provide energy for the cells.

Edited by QuestforLife, 08 July 2020 - 12:09 PM.

[Turnbuckle]

Given that valine, threonine, methionine and isoleucine feed into the krebs cycle at succinyl-coA, the step between AKG and Succinate, we might now want to avoid those amino acids if we really believe the succinate:AKG ratio is what is important in maintaining pluirpotency. Or might it be that such AAs relieve fatigue during the protocol precisely because they boost succinate? 

 

Leucine however increases the conversion of glutamate to AKG, so might be desirable. 

 

https://link.springe...0726-016-2254-8

 

I've found that methionine and lysine are necessary during both fusion and fission, and will soon add glutamine on a trial basis. Once the SC children are born, you gotta feed them, and they love methionine and lysine.

[genX]

Is the stearic acid portion of your protocol even safe? 

 

Here's a link to harmful and even toxic effects of stearic acid when ingested:   https://www.tandurus...id-effects.html

 

I also found another website (webmd?) which mentioned contracting "pneumonitis" from stearic acid  (from inhalation?) and that some people have sensitivity.

 

On the other hand I eat a lot of chocolate (rich in stearic acid) and know that stearic acid is found naturally in a number of foods (beef, chocolate etc.)

Also, I found this website for "the croissant diet" using butter enriched with stearic acid:  https://fireinabottle.net/

 

Any comments?

 

 

 

[Turnbuckle]

Is the stearic acid portion of your protocol even safe? 

 

Here's a link to harmful and even toxic effects of stearic acid when ingested:   https://www.tandurus...id-effects.html

 

 

 

Food grade stearic acid does not have these problems with pesticides and so forth. Nor is it being used except in a very intermittent fashion. Do not be concerned by it.

According to the FDA--

 

 In accordance with 184.1(b)(1), the ingredient [stearic acid] is used in food with no limitation other than current good manufacturing practice. The affirmation of this ingredient as generally recognized as safe (GRAS) as a direct human food ingredient is based upon the following current good manufacturing practice conditions of use...

 

https://www.accessda...cfm?fr=184.1090

 

 

If you are nevertheless concerned about it due to this posting by "Nick" on a holistic healing site, by all means do not use it.

Edited by Turnbuckle, 09 July 2020 - 01:22 AM.

[genX]

I know about the FDA label as GRAS but still thought I should ask given the conflicting information available on various websites.

I'm sure that others may have similar questions.

Also, it's important to know the purity etc.  Where do you get your stearic acid from?

[hsibai]

I am in my mid 50s and, perhaps, at the age when I should be learning more about this and other similar ideas and topics. I have come across a recent paper that seems highly relevant to this protocol but I am not at a level of domain knowledge that I can derive any conclusions. Perhaps some of the more knowledgeable followers of this topic can have a look to see if this protocol can benefit from it.
https://doi.org/10.1...tcb.2019.12.001

 

Edited by hsibai, 09 July 2020 - 06:25 AM.

[Andey]

I am in my mid 50s and, perhaps, at the age when I should be learning more about this and other similar ideas and topics. I have come across a recent paper that seems highly relevant to this protocol but I am not at a level of domain knowledge that I can derive any conclusions. Perhaps some of the more knowledgeable followers of this topic can have a look to see if this protocol can benefit from it.
https://doi.org/10.1...tcb.2019.12.001

 

  Both sulforaphane and curcumin activates Nrf2 and decreases DNA damage. I ve read a paper somewhere that Nrf2 activation leads to a 30% less DNA damage and therefor decrease the rate of aging(if the hypothesis in the paper  you cited is correct). Its even without taking into account stem cell activation and senolityc properties of the protocol.

[Andey]

.

Edited by Andey, 09 July 2020 - 07:37 AM.

[QuestforLife]

  Both sulforaphane and curcumin activates Nrf2 and decreases DNA damage. I ve read a paper somewhere that Nrf2 activation leads to a 30% less DNA damage and therefor decrease the rate of aging(if the hypothesis in the paper  you cited is correct). Its even without taking into account stem cell activation and senolityc properties of the protocol.

 

On the subject of sulforaphane I did some self-experimentation recently that may be of interest to this thread.

 

I've made no bones in the past about the fact that the sulforaphane paper quoted in this thread (http://dx.doi.org/10...dox.2016.11.007) that showed such an impressive increase in mitochondrial fusion with a concentration of 50uM seemed very difficult to achieve in vivo. 

 

I decided to test this for myself, however.

 

After taking nicotinamide once a week for a while, I became very well acquainted with its most noticeable effect - a distinctive type of mild headache ~2 hours after dosing 2 grams (no ribose). This seemed very repeatable. I can't really describe the headache other than to say it was mostly mild, sometimes forcing me to scrunch my forehead for a second or two, but was most noticeable when I moved my head around quickly. I wonder if any others have experienced this? I normally take nicotinamide in the morning, at least a few hours before food. Eating food within this window seems to make the headache worse for ~ 1/2 hour, before dissipating it almost entirely (although sometimes it comes back weaker). 

 

So one day I took 2 grams of nicotinamide with 3 tabs of prostaphane; each tab has 10mg stabilised sulforaphane (no need for an conversion to an active form in digestive system). I noticed no headache whatsoever. I was slightly more tired than usual that evening and I wouldn't recommend the combination. But it does seem to suggest sulforaphane is effective for fusion at least in the form I took it.

 

Just an anecdote, but might be helpful to others. 

[Turnbuckle]

I am in my mid 50s and, perhaps, at the age when I should be learning more about this and other similar ideas and topics. I have come across a recent paper that seems highly relevant to this protocol but I am not at a level of domain knowledge that I can derive any conclusions. Perhaps some of the more knowledgeable followers of this topic can have a look to see if this protocol can benefit from it.
https://doi.org/10.1...tcb.2019.12.001

 

Your paper says --

 

Genome instability plays a significant role in the progression of aging and protecting our aging genomes is therefore of fundamental importance for healthy aging...

 

 

 

This hypothesis has a decades long history that drove the whole anti-oxidant craze, but anti-oxidants have taken us up a blind alley. They don't make you live longer. A better understanding of aging based on epigenetics is emerging and this protocol is directed to that. The epigenome is the collection of DNA and histone markers that turn genes on and off, and thus distinguish the 200 cell types in the human body from one another, even though the underlying DNA is identical. The epigenome collects damage at a rate some ten times faster than the underlying DNA and this damage detunes cells for their specific functions. Fortunately there is a "sell-by" date in cells -- the telomeres. Thus, getting rid of telomerically old cells and replacing them with SC-derived somatic cells with de novo epigenetics substantially eliminates both genetic and epigenetic damage. 

 

As for the  QuestforLife's sulforaphane experience, stearic acid does not cross the BBB, and thus I've included sulforaphane in the protocol. Nicontinamide and sulforaphane both pass the BBB and produce opposing effects on mitochondria there. The first drives fission and the second fusion, so you may end up with neither. I suggest you try adding ribose to the nicotinamide. Ribose is a scarce commodity in cells and a large dose of niotinamide may thus result in an acute deficiency as cells make NAD from nicotinamide and ribose. Can that produce a headache? Possibly. A few posters at Amazon have reported headaches from taking large amounts of ribose or nicotinamide separately.

 

Also note that a high fission state increases the production of free radicals, while conversely, a fusion state reduces free radicals. High levels of free radicals can definitely produce a headache. It's interesting to note that it works both ways: ROS can also drive mitochondria to a fission state, while low levels can drive a fusion state.

 

Hence, manipulating mitochondrial morphology towards fission by pro-oxidants or towards fusion by antioxidants...seems to be a viable therapeutic strategy in combating cancer and degenerative diseases, respectively. 

https://www.ncbi.nlm...les/PMC5789323/

 

 

Edited by Turnbuckle, 09 July 2020 - 10:34 AM.

[yz69]

Hi Turnbuckle,

 

Do you still do satellite cell renewal protocol these days?  Does weight training after N+R+Curcumin helps getting rid of senescence cells?

 

Thanks!

[Turnbuckle]

Hi Turnbuckle,

 

Do you still do satellite cell renewal protocol these days?  Does weight training after N+R+Curcumin helps getting rid of senescence cells?

 

Thanks!

 

 

Skeletal muscle cells are different than most other cells. For one thing they can be enormous, and for another they are typically multinucleated, and may have hundreds of nuclei. Satellite cells typically merge with them and supply new nuclei by cell fusion. The more difficult question is what happens to dysfunctional nuclei. These nuclei do not replicate and thus the rate of epimutations is low, but obviously not zero. According to at least one paper, they cannot be removed by apoptosis, so exactly how is it done? Or can it be done at all?

 

While there is substantial data reporting the presence of apoptotic nuclei within the tissue following atrophic insults, recent data suggests that these are not true myonuclei, but rather, condemned mononuclear cells that reside outside the muscle fiber. 

https://www.frontier...2018.01887/full

 

 

 

About half the population older than 80 suffers from sarcopenia, and at present there is no treatment. A senolytic treatment for removing defective nuclei from skeletal muscles would thus be of great value. Even without that, increasing the numbers of satellite cells will certainly be helpful, as they are known to decline with age. Leucine is known to promote proliferation and differentiation of satellite cells, at least in rats, and thus can be used with C60+fusion.

 

Leucine, one of the indispensable branched-chain amino acids, is an important regulator of muscle mass through the control of protein synthesis [30]. Recently, Averous, et al. found that leucine limitation prevents the differentiation of both mouse-derived C2C12 myoblasts and primary satellite cells [31]. Chen et al. demonstrated that leucine could promote proliferation of C2C12 myoblasts [32]. These findings suggest that leucine plays more biological functional roles beyond the fundamental role as a substrate for protein synthesis in skeletal muscle tissue. Here, we prove that leucine could promote proliferation of primary preterm rat satellite cells in a concentration dependent manner.

https://www.ncbi.nlm...les/PMC4446757/

 

Edited by Turnbuckle, 09 July 2020 - 05:26 PM.

[genX]

Turnbuckle,

       I've been thinking of trying your protocol but have a few questions and a comment:

 

1) Your protocol has two parts:  (1) Stem cell renewal and (2) senescent cell replacement   Given that I believe that my stem cells are depleted (I did not have a positive reaction to C60 oil and have undergone a number of stressors which tend to deplete stem-calls) could it make sense to start initially with Part 1 for a while before including the 2nd part?

 

2) Am I correct in remembering from previous posts that you do the protocol approximately once a week?

 

3) Your protocol was arrived at by a combination of logic (reading scientific publications on biological effects of different agents) and self-experimentation (trial-by-error).  However, one point appears to me to be

self-evident:  different people have different genetics/epigenetics and physical makeup.  As a result, while it's possible for this approach to work for other individuals, it seems quite likely that the optimal doses

might vary from individual to individual.  For example, the doses you recommend might be too high or too small for someone with a different genetics, or just simply higher or lower body-weight/height/metabolism. 

Do you have any comments on this, or know of examples of anyone else who has successfully followed your protocol?

 

4)  Do you have any comments on the use of adipose-derived stem-cells, or alternatively bone-marrow based stem-cells, or alternatively exosomes derived from placental cord-blood, as an alternative to part 1 of your protocol for stem-cell renewal?

 

5) There's been a lot of discussion recently about rejuvenation via simply eliminating toxins from blood-plasma or donating blood to force blood-plasma renewal or cleansing or replacing plasma with artificial plasma (Nugenics on this website) or replacing blood plasma with plasma from young individuals.  Do you see any connection between your protocol and any of these procedures?

 

 

 

[genX]

Not sure why no response to any of my queries. Perhaps there were too many? 

 

BTW, the entire protocol is based on Turnbuckle's assumption that C60 blocks UCP2.  I'm not

aware of any published evidence for this. 

 

Furthermore, from a personal point of view one part of the protocol (e.g. SAMe) clearly disagrees with me, most likely because

I am over-methylated. So, due to this perhaps I don't need a methyl donor to implement this protocol?

[Turnbuckle]

 

BTW, the entire protocol is based on Turnbuckle's assumption that C60 blocks UCP2.  I'm not

aware of any published evidence for this. 

 

 

 

My UCP2 hypothesis was discussed 2 years ago and you can find it on the first 2 pages of this thread. It is an explanation for how C60 stimulates stem cells, for there is plenty of evidence that it does. 

[p75213]

It seems sulforaphane also stimulates stem cells:-
"SFN of low concentrations stimulated cell proliferation and prominently increased neurosphere formation and NSC differentiation to neurons. SFN treatment upregulated Wnt signaling in the NSCs."

https://pubmed.ncbi....h.gov/28142224/

[Edit_XYZ]

Stem cells are activated by hepatocyte growth factor activator (HGFA). Normally, injury activates stem cells by HGFA, but the effect is also achieved by directly injecting HGFA:

https://www.cell.com...1247(17)30427-8

[eighthman]

https://www.fightagi...l-age/#comments

 

I would think that the body places a priority on the heart, as to survival.  One quote is interesting as to a zero age in some cardiac stem cells.

[Turnbuckle]

https://www.fightagi...l-age/#comments

 

I would think that the body places a priority on the heart, as to survival.  One quote is interesting as to a zero age in some cardiac stem cells.

 

There's nothing mysterious about it. Organs age at different speeds. Some may even age in reverse from time to time (as when one gives up smoking and drinking, or joins a gym). The rate of replacement of epigenetically old somatic cells with stem cells having very low epigenetic age will determine this. Filling stem cells niches while clearing out senescent cells will synergistically lower overall age, with some organs benefiting more than others. 

Edited by Turnbuckle, 21 July 2020 - 12:34 PM.

[orion22]

There's nothing mysterious about it. Organs age at different speeds. Some may even age in reverse from time to time (as when one gives up smoking and drinking, or joins a gym). The rate of replacement of epigenetically old somatic cells with stem cells having very low epigenetic age will determine this. Filling stem cells niches while clearing out senescent cells will synergistically lower overall age, with some organs benefiting more than others. 

what to do you think about combine c60 with allot of hydrogen gas

here is a study of effects of hydrogen on multipotential stromal cells 

https://pubmed.ncbi....h.gov/20570654/

[Turnbuckle]

what to do you think about combine c60 with allot of hydrogen gas

here is a study of effects of hydrogen on multipotential stromal cells 

https://pubmed.ncbi....h.gov/20570654/

 

The results of this paper are positive, but not earthshaking, even in vitro. For one thing, the exposure was typically for weeks, and for another, the 3% H2 used is just below the flammability limit. 

[Starjumper7]

It is said in this thread that stearic acid induces fusion, and in the mitochondria fission/fusion thread it states that glycerol monostearate induces fusion, so it looks like they are interchangeable.  My question is, what is the ratio of effectiveness, is 10g of stearic acid equal to 5g of glycerol monstearate?

[Turnbuckle]

Update on my epigenetic age.

The last 4 tests from TruMe --

 

Date      Epigenetic age - Chronological age, years

02/2018.... +0.5 (Baseline before treatments)

11/2019.... -13.0

02/2020.... -14.2

04/2020.... -14.5

06/2020.... -13.0

 

I was disappointed to see my epigenetic age rise with the last test. In this two month period I added one gram of cinnamon powder to the senolytic treatments, in the expectation that it would shorten telomeres of replacement cells, increasing cellular throughput and thus decrease epigenetic age faster. There isn’t much  cinnamaldehyde content in the powder, however, with one gram containing a variable amount of 7-31 mg. After sending in that last sample, I obtained purified materials and began using 50 mg of cinnamaldehyde and 50 mg of eugenol in place of the powder. The subjective effects are good, but we’ll have to see with the next TruMe test.

 

For references and more on the rationale of using telomere shortening agents, see my post #1206.

 

Note: I added the cinnamaldehyde and eugenol to a like amount of olive oil and mixed this into fruit juice with a NutriBullet. Otherwise the taste would be highly objectionable.

 

 

[stephen_b]

The error rate is stated on their website as 4.75 years, so it's hard to say for sure exactly what the change was. I wouldn't get too disappointed given the error bars.

[yz69]

Hi Turnbuckle,

Did you have any infection that could cost some young WBCs during the two month period? Have your done any CBC tests?

[yz69]

I have tried Turnbuckle's protocol for 6 months, doing one round per week. I am 50, my baseline Trume age is 47.5. I have seen my Trume age went up to 50.2 in February, probably due to my sore throat for a week (possible infection?) in that month.  After that, Trume age started to drop, to 44.9 in June.  My gray hair has not changed, my look has not changed, the biggest change comes from the white blood cells:

 

 

February:

Neutrophils (Absolute)   2.1

Lymphs (Absolute)   2.3

Monocytes(Absolute) 0.5

 

June:

Neutrophils (Absolute)   1.9

Lymphs (Absolute)   3.7

Monocytes(Absolute) 0.5

 

How to explain the jump in lymphocytes and drop in neutrophils?

 

[Andey]

Probably infection of some sort. Though its strange as I was under the impression that lymphocyte population is much more stable than neutrophils. Neutrophils could buildup population really fast in a response to the infection, lymphocytes need to undergo a long process of positive and negative selection, meaning that your increase took at least a month.

 Drop in neutrophils is not meaningful as its not really dropped significantly, its within a range of natural variation.

Edited by Andey, 30 July 2020 - 06:08 AM.

[yz69]

Got the latest Trume result back, now trume age becomes 43. Also the latest blood test in July still shows drops in  neutrophils  and relative high  lymphocytes:

 

July:

Neutrophils (Absolute)   1.5

Lymphs (Absolute)   3.3

Monocytes(Absolute) 0.5

 

I was just doing Turnbuckle's protocol, nothing else.

[yz69]

According to this paper, https://academic.oup...irectedFrom=PDF

 

 

Analysis of 1684 inpatient and 884 outpatient peripheral blood counts revealed the following: (1) The absolute lymphocyte count declines sharply from 5000/μ,l to 2000/μl in the first two decades, remains constant for three decades, then declines at an accelerated rate beginning in the 40s, to reach 1500/μl at age 90.

Maybe the increase of absolute lymphocyte count is a sign that I am aging backward?

 

Or could it be we just have more stem cells flowing in the blood, however counted as lymphocytes?

 

 

 

Lymphocytes vary widely in size. Small lymphocytes are 7 to 10 µm in diameter, and large lymphocytes are approximately 14 to 20 µm in diameter, although intermediate sizes may be encountered.

 

CD34+ stem cells looks like lymphocytes but they are smaller , according to this paper

 

  https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.b.20399#

 

 

 

 

Results: Lymphocytes, monocytes and granulocytes in the HPC-A samples had a mean electronic volume of 414, 797, and 670 um3 , respectively corresponding to cell diameter of 9.25, 11.5, and 10.85 um. In 53 HPC-A samples analyzed, the mean electronic volume of the CD34 positive mononuclear cells was 407 um3 while the CD45 positive cells had mean volume of 453 um3 .

 

 

 

The drop in neutrophils could be the result of curcumin:

https://www.ncbi.nlm...les/PMC4687759/

 

 

Herein, we confirmed that curcumin induces human neutrophil apoptosis as assessed by cytology and by increase in the cell surface expression of annexin-V and CD16 shedding. 

 

Edited by yz69, 30 July 2020 - 05:53 PM.

[dlewis1453]

Got the latest Trume result back, now trume age becomes 43.

 

This is very exciting! Now we have another case of an individual reversing their epigenetic age with Turnbuckle's stem cell protocol.