2523 replies to this topic

[Andey]

Reading over "Stem cell aging: mechanisms, regulators and therapeutic opportunities" brings to mind Hector Zenil's work on complex systems modeling.  Specifically, there are systematic approaches facilitating rapid convergence toward predictive models opened up by advances in the application of algorithmic information theory based on sparse datasets.

A simple example would be compiling a dataset of longitudinal measures and experimental outcomes, and constructing stock and flow diagrams to represent various models.  Model selection then consists of measuring the degree to which the models losslessly compress the dataset -- including the models themselves as aspects of the respective decompression programs.  I know this seems alien to most scientists but it is entirely justified as a model selection criterion.  If you think the dataset is impoverished in some way, do the appropriate measurements and add the data to the dataset.

 

 Some seriously smart people amongst us))

 

  If I caught the gist of it, its the same as neural network works - it linearly regresses with training to the most suitable yet the most simple model. I would not say its THE correct way thought as models often fail with new data coming, but its simple and practical, that's for sure.

 

To avoid complete offtopic:

I think there is some synergy with the protocol and hydrogen water. Usually, my HRV plummet the night after leg exercise day. If it coincides with C60 than fall is less pronounced, but when its C60 + hydrogen water than my Oura ring shows completely normal HRV during the night.

Edited by Andey, 25 March 2019 - 10:51 AM.

[bosharpe]

 

Proliferation is obtained by symmetric division, whereas differentiation is asymmetric. When I began using C60 back in 2012, I didn't appreciate C60's mode of action, that C60 was stimulating stem cells. If you don't take care to proliferate them, however, you can use them up. In fact, that is the normal situation with aging--

 

 

 

 

There are many causes of aging, but in my view the most central one is the Hayflick crisis that occurs with everyone. Large numbers of somatic cells reach senescence, apoptosis mechanisms get behind, and replacement is limited by depletion of the stem cell pools. So the goal here is to increase both the stem cells pools and apoptosis.

 

 

Yeah, same here. I was too hasty and foolish when I tried C60oo for a short period a few years ago, without this knowledge. 

[jabowery]

...
  If I caught the gist of it, its the same as neural network works - it linearly regresses with training to the most suitable yet the most simple model. I would not say its THE correct way thought as models often fail with new data coming, but its simple and practical, that's for sure.

Well, its more general than NNs in that NNs -- particularly recurrent (feedback loops) NNs -- can represent stock and flow diagrams, and vise versa. You can avoid recurrence to first order by resorting to path analysis but that's just to get started toward a recurrent model. You can't really model dynamical systems without recurrence (feedback loops).  

As for new data coming in, data compression operates by prediction. If you can predict the next bit in a bit stream, you don't need to store it. If the next bit is not what the algorithm predicted, then it is stored. That's how you measure how "predictive" your model is with lossless compression.

To avoid complete offtopic:...

My apologies for "derailing" the current thread with a meta issue. Where does this belong?

Edited by jabowery, 26 March 2019 - 04:07 PM.

[Fafner55]

Guarente found success in proliferating stem cells and regenerating the gut in aged mice with large doses of NR over 6 weeks. 

 

“Biologists find a way to boost intestinal stem cell populations" (2019) https://medicalxpres...-stem-cell.html

Cells that line the intestinal tract are replaced every few days, a high rate of turnover that relies on a healthy population of intestinal stem cells. MIT and University of Tokyo biologists have now found that aging takes a toll on intestinal stem cells and may contribute to increased susceptibility to disorders of the gastrointestinal tract.

The researchers also showed that they could reverse this effect in aged mice by treating them with a compound that helps boost the population of intestinal stem cells. The findings suggest that this compound, which appears to stimulate a pathway that involves longevity-linked proteins known as sirtuins, could help protect the gut from age-related damage, the researchers say.

"One of the issues with aging is organ dysfunction, accompanied by a decline in the activity of the stem cells that nurture and replenish that organ, so this is a potentially very useful intervention point to either slow or reverse aging," says Leonard Guarente, the Novartis Professor of Biology at MIT.

 

"NAD+ supplementation rejuvenates aged gut adult stem cells" (2018) https://onlinelibrar...111/acel.12935 

The tissue decline due to aging is associated with the deterioration of adult stem cell function. Here we show the number and proliferative activity of intestinal stem cells (ISCs) but not Paneth cells decline during aging, as does ISC function assessed ex vivo. Levels of SIRT1 and activity of mTORC1 also decline with aging. The treatment with the NAD(+) precursor nicotinamide riboside (NR) rejuvenates ISCs from aged mice and reverses an impaired ability to repair gut damage. The effect of NR is blocked by the mTORC1 inhibitor rapamycin or the SIRT1 inhibitor EX527. These findings demonstrate that small molecules affecting the NAD/SIRT1/mTORC1 axis may guide a translational path for maintenance of the intestine during aging.

 

[Turnbuckle]

 

Guarente found success in proliferating stem cells and regenerating the gut in aged mice with large doses of NR over 6 weeks. 

 

 

 

Have these effects been noted systemically? If not, it is yet more evidence that NR is digested prior to absorption. As for the 6 week time frame, C60 looked good over the lifetime of rats, but seemed to fade in humans over a period of years, for the reasons that have been discussed from the opening post. So let's please not have NR intruding into this thread. It's a false hope, driven by marketing.

[Fafner55]

At the minimum, Guarente's publication shows that significant regeneration is possible and possible pathways involved. But while he mentions the relationship between NAD+ and mitochondrial defects, it is surprising that he doesn't mention mitophagy. Also, there is no discussion of inflammation that may be related to leaky gut. I don't know Guarente's reasons for keeping the scope of this research narrowly focused, but it does raise questions.

 

Contrast Guarente's publication with the impressive regeneration of the gut demonstrated in a recent publication by Longo. With high turnover tissues like the gut, promoting proliferation alone appears to be sufficient to achieve regeneration.

"Fasting-Mimicking Diet Modulates Microbiota and Promotes Intestinal Regeneration to Reduce Inflammatory Bowel Disease Pathology" (2019) https://www.cell.com...1247(19)30181-0  

[Fafner55]

Turnbuckle, regarding your post #720, is there evidence that the Hayflick limit can be extended in a meaningful way by eliminating senescent cells? It seems reasonable but I haven't seen published results.

[Turnbuckle]

Turnbuckle, regarding your post #720, is there evidence that the Hayflick limit can be extended in a meaningful way by eliminating senescent cells? It seems reasonable but I haven't seen published results.

 

The Hayflick limit refers to individual cells. The cellular Hayflick limit can be exceeded by telomerase activators, but I believe it’s generally unwise to do so, as this allows cells to grow epigenetically older than would otherwise be possible. Better to kill and replace.

[mkutsen]

The Hayflick limit refers to individual cells. The cellular Hayflick limit can be exceeded by telomerase activators, but I believe it’s generally unwise to do so, as this allows cells to grow epigenetically older than would otherwise be possible. Better to kill and replace.

It should be noted that in adults even stem cells have shorter telomers.  Thus they are not capable of as many divisions as embryonic stem cells.  

When inducing pluripotent stem cells the telomers revert  back to the original length.

[mkutsen]

It should be noted that in adults even stem cells have shorter telomers.  Thus they are not capable of as many divisions as embryonic stem cells.  

When inducing pluripotent stem cells the telomers revert  back to the original length.

Here are some references:

1. https://www.ncbi.nlm...pubmed/21222203

 

2. https://www.ncbi.nlm...les/PMC2360127/

Edited by mkutsen, 06 April 2019 - 03:10 AM.

[Turnbuckle]

It should be noted that in adults even stem cells have shorter telomers.  Thus they are not capable of as many divisions as embryonic stem cells.  

When inducing pluripotent stem cells the telomers revert  back to the original length.

 

Embryonic stem cells are the key. The typical progression goes from embryonic SCs Þ adult SCs Þ transit amplifying cells Þ somatic cells.

 

Embryonic (pluripotent) cells weren't known to exist in the adult until a few years ago, and apparently escaped detection due to their small size, and thus they are called Very Small Embryonic-Like Stem Cells (VSELs).

 

This evidence supports that VSELs are pluripotent stem cells that most likely serve as precursors of both mesenchymal and hematopoietic compartments of stem and progenitor cells. 

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

 

Pluripotent VSELs ... were first reported by Ratajczak and group in adult mice tissues ... These are diploid cells with high telomerase activity...

https://www.ncbi.nlm...cles/PMC3586435

 

 

Thus I made these cells the primary target of this protocol, using threonine because that is their primary nutrition--

 

Embryonic stem cells (ESCs) undergo unlimited self-renewal while maintaining a pluripotency, which is defined as the ability to develop into cells of all three embryonic germ layers. ESC self-renewal is characterized by special proliferative and epigenetic properties and a unique metabolic profile. One of the key features of this specialized nutritional metabolism is a stringent requirement for the amino acid threonine.

https://www.ncbi.nlm...pubmed/24232288

 

Mouse embryonic stem (mES) cell proliferation depends exclusively on the nutritionally essential amino acid, L-threonine, in the medium. Other essential and non-essential amino acids need not be added to the medium for mES cell proliferation. 

https://www.ncbi.nlm...les/PMC4206991/

 

 

One could, of course, take telomerase enhancers to lengthen the telomeres of adult stem cells, but they also lengthen the telomeres of transit amplifying cells, and that blocks the turnover you need to reverse epigenetic age as it allows those transit amplifying cells to get epigenetically older than they would normally. Early on I worried about telomere shortening of adult stem cells and added a telomerase enhancer to the protocol, only to see my initial drop in epigenetic age begin to rise again. It was then I realized that you don't want to mess with telomeres in a global sense, as they are the sell-by dates by which potentially old and dysfunctional cells are eliminated.

[Vany]

 

2. It is intended as a geriatric treatment, not for young people.

 

 

Hi Turnbuckle,

 

Thank for your great contributions here and for this protocol. After reading this thread I still don't understand why this protocol would not be beneficial for young people (~30-40 yo). Wouldn't it prevent aging by preserving a good stem cell pool ? Can you elaborate on this please ? Thank you !

[Rocket]

Hi Turnbuckle,

 

Thank for your great contributions here and for this protocol. After reading this thread I still don't understand why this protocol would not be beneficial for young people (~30-40 yo). Wouldn't it prevent aging by preserving a good stem cell pool ? Can you elaborate on this please ? Thank you !

 

Wouldn't it prevent aging? No. Stem cells are just one small part of the aging process. Stem cells would help with healing wounds. But stem cells do not work well in an aging extracellular matrix.... therefore even in wound healing, in the "old" there would likely not be much of a benefit. It also wouldn't do anything for sarcopenia to maintain youthful muscle in your later years. 

 

Before you start trying to quantify this as a method to reverse/hault aging, you need to first quantify how stem cells diminish with the years (actual real numbers) and how many stem cells this protocol "creates" (again, real quantifiable numbers).

 

Someone 30-40 should be focused on nutrition, maintaining low body fat, exercise, rest (often overlooked and underestimated) and perhaps telomeres and mitochondrial health.

[Turnbuckle]

 

 

Someone 30-40 should be focused on nutrition, maintaining low body fat, exercise, rest (often overlooked and underestimated) and perhaps telomeres and mitochondrial health.

 

 

I've stated several times that this is not a protocol for young people as there are too many unknowns. If you are in your thirties, you have plenty of time to let the geriatric crowd experiment with stem cells As for mitochondrial health, I've addressed that on another thread--Manipulating mitochondrial dynamics, with a protocol here--whereby the mito populations of cells can be returned to more youthful levels. I don't see any reason why young people should not use it, except having substantial defective mitochondria at that age would likely be rare. As for telomeres, they should not be tinkered with at all, in my opinion. These are the expiration dates on cells, thus it's not surprising that longer telomeres are associated with accelerated epigenetic aging, and epigenetic age closely matches chronological age. See more at Reversing the Clocks.

Edited by Turnbuckle, 24 April 2019 - 06:38 PM.

[Engadin]

Some AOVEs in Spain, my country, have the world record in polyphenol content, such as 2.115 mg/kg.

 

The amount of oleocanthal in the same AOVE brand is 812 mg/kg.

 

ECOLIBOR Certificate of Analysis: http://chrome-extens...2/Prokopios.pdf

 

ECOLIBOR webpage: https://www.ecolibor...re-polyphenols/:

 

"Polyphenols are naturally occurring chemicals that are present in various vegetables. What makes them so special, though, is their potential positive impact at an antioxidant and cardiovascular level. Currently, scientific studies are focusing on one of these polyphenols in particular, oleocanthal, which has tremendous anti-inflammatory properties. The Oleocanthal “in vitro” laboratory, for example, has shown that the antioxidant is able to kill cancer cells without harming healthy human cells. This is the conclusion of a recent study conducted by a team of scientists at Rutgers University in New Jersey, and Hunter College in New York, which was published in Journal of Molecular and Cellular Oncology (Feb 2015) .

 

Emphasizing this point, Olive Oil Times wrote about a study using a sample of 110 oils and using the same analysis system as we use for our oil. The result? The oil with the highest polyphenol count was a California brand with a total of 1232.7 mg/kg — as compared to ours, which has a count of 2.115 mg/kg (see our analysis here for more details).

 

EFSA (European Food Safety Authority) has approved the use of the following nutrition declaration for polyphenols in olive oil (Commission Regulation (EU ) 432/2012) : “The olive oil polyphenols contribute to the protection of blood lipids from oxidative damage. This statement can be placed only on oil containing at least 5 mg of hydroxytyrosol and its derivatives per 20 g of olive oil.” To achieve this effect, consumer’s should be notified that these benefits are obtained with a daily intake of 20 g of oil. In the case of our olive oil, according to the current chemical analysis, we have 36mg of hydroxytyrosol derivatives per 20g."

 

My guess is this AOVE is an outstanding candidate for our C60 mix, adding remarkable properties from abundant polyphenol, oleuropein (333 mg/kg.), hydroxytyrosol and oleocanthal compounds.

Edited by Engadin, 30 April 2019 - 10:58 PM.

[dlewis1453]

This thread has inspired me to research the senolytic FOXO4, since removing senolytic cells is the complementary therapy to the stem cell renewal protocol. 

 

I found an interview with Dr. Peter de Keiser, the main researcher into FOXO4, at the Life Extension Advocacy Foundation, that was very insightful. 

 

In this interview, Dr. Keiser said that FOXO4 is a very powerful and selective senolytic substance, but that it still kills 1 non-senescent cell for every 9 senescent cells that it kills. He viewed this is a downside to the therapy, but it occurred to me that if we are able to create new stem cells with Turnbuckle's protocols, this might not be a problem. 

 

Perhaps the 1 out of 10 normal cells that are being killed by FOXO4 are actually cells that are nearing senescence, and should be replaced by new cells anyway. Maybe FOXO4 could be effective when paired with Turnbuckle's protocol. 

 

By the way, I found interesting positive anecdotal reports on FOXO4 across the internet, such as at the FightAging website. I should also note that the old mice treated with FOXO4 experienced strong rejuvenation. Imagine then what could be accomplished with FOXO4 married with stem cell self-renewal. 

[Turnbuckle]

 Maybe FOXO4 could be effective when paired with Turnbuckle's protocol. 

 

 

 

 

Possibly, but it has to be injected, right? And it's rather expensive. My goal for all protocols is to make them cheap and easy to use.

 

I'm beginning to believe that the senolytic part of this protocol is not very difficult when combined with the stem cell treatment. Senescent cells are known to put out a senescence associated secretory phenotype, and it's my hypothesis that stem cells put out something similar. This SC secretory phenotype hypothetically gives senescent cells permission to commit suicide. And that seems like a good idea, for does the body want senescent cells to commit suicide if there is nothing available to replace them? That might leave holes in tissues that would prove fatal. So by this hypothesis, once stem cell pools are filled, apoptosis of senescent cells is more readily achieved, which seems to be the case in my experience.

[dlewis1453]

Possibly, but it has to be injected, right? And it's rather expensive. My goal for all protocols is to make them cheap and easy to use.

 

I'm beginning to believe that the senolytic part of this protocol is not very difficult when combined with the stem cell treatment. Senescent cells are known to put out a senescence associated secretory phenotype, and it's my hypothesis that stem cells put out something similar. This SC secretory phenotype hypothetically gives senescent cells permission to commit suicide. And that seems like a good idea, for does the body want senescent cells to commit suicide if there is nothing available to replace them? That might leave holes in tissues that would prove fatal. So by this hypothesis, once stem cell pools are filled, apoptosis of senescent cells is more readily achieved, which seems to be the case in my experience.

 

Yes that is correct, it has to be injected, which is certainly inconvenient. It is definitely more expensive than say fisetin or quercetin, but not bad when compared to many pharmaceuticals, especially since the dosages are small. On its own it appears to be much more powerful than any other senolytic when it comes to killing L-6 type senescent cells, but it is not effective against MNP3-high senescent cells. This is something I learned recently about senolytics - that there are different types of senescent cells, and most senolytics are not effective against all of them, therefore taking a variety of senolytics is necssary. Different senolytics may also have synergies between each other. 

 

What you just wrote regarding the interaction between the replenished stem cell pools and senescent cells is very interesting and also encouraging. Your theory makes sense, and if true it would mean that oral herbal senolytics may be all we need. In that case, powerful senolytics like FOXO4 may only be useful if someone needs extra help at the beginning to clear out an especially large burden of senescent cells. 

[ambivalent]

Would there not be exepcted benefit from just taking stearic acid over a period of time, if the stem cell pools are recovered wouldn't the body just natrually utilise this stem cell boon albeit protractedly without the need for c60?  Additionally, might it be beneficial to take a significant course of stearic acid before embarking on senolytic protocol? It would seem prudent to ensure there is a burgeoning stem-cell pool before wiping large numbers of sensescent cells.

[Turnbuckle]

Would there not be exepcted benefit from just taking stearic acid over a period of time, if the stem cell pools are recovered wouldn't the body just natrually utilise this stem cell boon albeit protractedly without the need for c60?  Additionally, might it be beneficial to take a significant course of stearic acid before embarking on senolytic protocol? It would seem prudent to ensure there is a burgeoning stem-cell pool before wiping large numbers of sensescent cells.

 

You don't want to take stearic acid continuously. To do so would keep your mitochondria in fusion, which would result in the build up of defective mtDNA as the cellular quality control mechanism requires fission. When stem cell pools are full, then you can switch to a mostly senolytic protocol and tissues will use the available stem cells to make replacements (which for geriatrics, will be required at a much higher rate than when you were young). To use those stem cells requires asymmetric division of stem cells, which requires a non-fusion state of mitochondria. Apoptosis also requires a non-fusion state, thus a fission state works best for a senolytic protocol.

 

Prior to, or simultaneous with, cytochrome c release and upstream of caspase activation, mitochondria fragment into multiple small units. Blocking of this mitochondrial fission inhibits cytochrome c release and delays cell death.

https://www.ncbi.nlm...les/PMC2732420/

 

[ambivalent]

This article implies that the benefit of fasting resulted from mitochondria being maintained in a fused state but also suggests one-state mitochondria is, as you infer, unhealthy: “Our work shows how crucial the plasticity of mitochondria networks is for the benefits of fasting. If we lock mitochondria in one state, we completely block the effects of fasting or dietary restriction on longevity.”

 

https://scitechdaily...rease-lifespan/

 

But if fasting and ketogenic diets, which we know to be beneficial, are maintaining mitochondria in a fused states for long periods of time then will taking SA for a reasonable period to build up stem cells be so harmful? In addition, is there any indication of how quickly the stem cell pool can be repleted?

Edited by ambivalent, 26 May 2019 - 11:49 AM.

[Turnbuckle]

This article implies that the benefit of fasting resulted from mitochondria being maintained in a fused state but also suggests one-state mitochondria is, as you infer, unhealthy: “Our work shows how crucial the plasticity of mitochondria networks is for the benefits of fasting. If we lock mitochondria in one state, we completely block the effects of fasting or dietary restriction on longevity.”

 

https://scitechdaily...rease-lifespan/

 

But if fasting and ketogenic diets, which we know to be beneficial, are maintaining mitochondria in a fused states for long periods of time then will taking SA for a reasonable period to build up stem cells be so harmful? In addition, is there any indication of how quickly the stem cell pool can be repleted?

 

A worm that lives for 2 weeks can tell us little about the effect of mito fission/fusion on human longevity.

[ambivalent]

No but I'm interested to know if fusion is a fixed state of fasting/ ketosis - which the article suggests then appears to contradict - which we know is pretty healthy for human longevity. 

[Turnbuckle]

No but I'm interested to know if fusion is a fixed state of fasting/ ketosis - which the article suggests then appears to contradict - which we know is pretty healthy for human longevity. 

 

 

This thread is about attacking the core driver of aging--epigenetic mutations. Lowering one's burden of epigenetic mutations by replacing epigenetically old somatic cells with epigenetically young stem cells. Fasting doesn't play a part in it.

[ambivalent]

I understand but this diversion resulted from query as to whether or not taking SA for a extended period of time was viable in order to build up stem cell supplies, pre-senolytic. You objected on the grouds that lengthy mitochondrial fused states would be harmful - the article appears to suggest that fasting might induce extended periods of mito-fusion, if true that would make me less concerned about taking it a few weeks at a time, say. But as I said it seems rather contradictory, as mentioned earlier. 

 

Additionally, if fasting clears out dead cells why couldn't it play a role in stem-cell replacement? Anyhow, I'll leave the diversion there. 

[Rocket]

This thread is about attacking the core driver of aging--epigenetic mutations. Lowering one's burden of epigenetic mutations by replacing epigenetically old somatic cells with epigenetically young stem cells. Fasting doesn't play a part in it.

I thought the primary driver of aging was gene expression, whereas stem cell activity was diminished in aging tissues because of signaling, senescent cell accumulation, and epigenetic changes to tissue inhibiting stem cells. I thought your stem cell protocol was so something akin to a "brute force" or a way to trick the bodies tissues to promote stem cell activity.

Obviously there are many driving factors responsible for aging, but I thought consensus was pointing toward gene expression for the primary cause. By primary, I mean that if you were to look at the causes on a pie chart, that gene expression would be the largest slice of "pie".

[Turnbuckle]

Obviously there are many driving factors responsible for aging, but I thought consensus was pointing toward gene expression for the primary cause. By primary, I mean that if you were to look at the causes on a pie chart, that gene expression would be the largest slice of "pie".

 

What do you think controls gene expression? It's epigenetics.

[Engadin]

What do you think controls gene expression? It's epigenetics.

 

Not only:

 

 

 

[ambivalent]

Maintaining the fasting/fusion detour a little longer: out walking today I wondered why do stem cells divide symmetrically under mitochondrial fusion? The has not been discussed, as far as I am aware.

 

It seemed quite obvious. Many of us know from experience the regenerative properties of fasting; indeed Longo observed that white blood cell counts drop off during fasting, as old cells are recycled for energy, but spike on refeeding (above pre-fasting levels) - the cause? Stem cells. To borrow from Cynthia Kenyon, we clearly possess this latent capacity to live longer under CR (and or possibly fasting), so fasting would of course represent something of a short-term gain if stem-cell pools were depleted during tough times, we'd wind up burning twice as bright but half as long, so symmetric differentiation would allow the body to service its need for rejuvenation during lean times but also retain a priori levels of stem cells. Additionally, it would make a lot of evolutionary sense for mitochondria  to fuse during fasting since, depleted of energy, a fusion boost would represent a critical advantage. The mitochondrial fused state then presumably signals symmetric differentiation.     

 

If true, then it shouldn't be a problem to retain a fused state for a few weeks at a time (unless say there is some periodic forced fission during fasting to maintain the plasticity mentioned important in the article a few posts back). Certainly, there are peaks and troughs in energy-levels during extended fasts. 

Edited by ambivalent, 28 May 2019 - 10:01 PM.

[ambivalent]

Re refernces - the mitochondrial fasting fusion state is referenced further in the thread; the depletion of stem cells during fasting, in the post; the increased output from mitcochondrial-fusion much referenced in threads. Making a hypothesis based on these established observations doesn't require a reference: that's the point, its a hypothesis. Yes downvoting is empowering, but a little more thought before making casual judgements would be appreciated by all, better still written engagement - otherwise the precise nature of the objection is less than clear.