2503 replies to this topic

[userCK]

This is tangential to the SC protocol but I figured I might as well ask here. 

Is there a way to find out if NOS genes have been methylated in my body? From Wiki:

• NOS3 or eNOS gene appears in Chromosome 7 (7q35-7q36)
• The gene coding for nNOS is located on Chromosome 12

 

I am wondering if it is possible that these genes responsible for producing Nitric Oxide Synthase enzymes has been methylated? And if so, perhaps this SC protocol can somewhat reverse it by adding more stem-cells with denovo methylation? If so, then body should start producing Synthase enzymes again. And if that happens, then I should start responding to Cialis, Viagara, or Nitric Oxide supplements like L-Arginine/L-Citrulline.

 

Also, a potential test could be taking Synthase. If Synthase is missing in my body and I take it and then take L-Arg/L-Citrulline + Cialis and it works, then that means these genes are methylated? But does anyone know if it is possible to procure and inject 'Synthase'?

 

EDIT:

https://www.nature.c...cles/tpj201149 

patients with erectile dysfunction carrying the C allele for g.-786T>C polymorphism showed better responses to PDE-5 inhibitor sildenafil

My genotype is T/T (according to 23andMe data explorer of my sequence, SNP: rs2070744)

So, it seems to me I was already less likely to respond to Cialis/Viagra and then after taking Finasteride/Minoxidil/Accutane, I may have methylated NOS3 genes and that pretty much killed the production of this synthase enzyme/nos3 enzyme. So, if I find a way to demethylate it, then that problem would go away. 

 

Edited by userCK, 18 February 2021 - 10:10 PM.

[userCK]

@Turnbuckle - could you suggest a protocol for demethylating brain cells? I've read that TET1 enzyme (which can be upregulated by Estradiol) can demethylate neuronal cells . Lithium is also a HDAC inhibitor and crosses blood brain barrier.

 

So, something like this protocol:

1. Testosteorne cream/injection to increase Estradiol. Not too high testosterone because that'd cause too high of estrogen. So would need to experiment with dosage. As I was on TRT for over a year, I think for me, 0.25ml of 20% 200mg/1ml cream would be sufficient. My prescription was for 1ml a day but that required me to take Anastrazol and that'd negate the goal of upregulating TET1.

 

2. Lithium Ortotate 50mg

 

Combined, over time, this should demethylate neuronal cells. But compared to your level of knowledge, I know nothing. So, I'm wondering if you've any other advise/suggestion?

 

SC protocol would demethylate in body and I think that might (over time) resolve NOS3 issues by demethylarting NOS3 genes. But nNOS is produced in brain and my guess it would require me to demethylate neuronal cells somehow.

 

 

[userCK]

Hi Turnbuckle,

 

Can we add HGH (human growth hormone) to this protocol and would that accelerate epigenetic reversal? If so, where to add it ? Part 1, Part 2? 

[Danniel]

@Turnbuckle: With age, many of us suffer from different autoimmune diseases. What would be the possible effects of each phase of this regimen on an autoimmune disease?

[Turnbuckle]

@Turnbuckle: With age, many of us suffer from different autoimmune diseases. What would be the possible effects of each phase of this regimen on an autoimmune disease?

 

 

SC pool expansion is potentially helpful where aging results in senescence of the immune system. See Adipose-Derived Mesenchymal Stem Cells in Autoimmune Disorders: State of the Art and Perspectives for Systemic Sclerosis --

 

7 CONCLUSION AND PERSPECTIVES

Taken together, the work carried out in the last decade demonstrated that MSC might

represent an innovative strategy to cure AID. In particular, MSC displaying

immunosuppressive, anti-fibrotic, pro-angiogenic and anti-oxidative responses,

40

harbor new hope for the treatment of SSc, a multifaceted intractable AID with unmet

medical need. While a first clinical trial using MSC in SSc has been launched in

France, results obtained in preclinical models, as well as the few case reports in the

human disease are very promising. Considering that MSC mainly act through a “hit

and run” mechanism, involving paracrine, endocrine and extracellular vesicles

secretion, the use of allogeneic MSC seems a reasonable setting to treat AID, where

resident MSC might be impaired and even contribute to disease progression.

Regarding the source for MSC, the current knowledge prompts to investigate diverse

sources of MSC, among which adipose tissue is highly promising. In that context, the

convincing effects obtained with ASC in the HOCl preclinical model, and in other AID,

are particularly appealing for the treatment of SSc. However, further studies will have

to focus on better characterization of MSC/ASC functionality and the development of

potency assays, in order to individualize cell-therapy according to patient’s needs,

and develop relevant randomized controlled trials in SSc. 

 

 

and T Cell Senescence and Autoimmunity --

 

Transcriptome profiles of the SA-T cells of both aged B6 and female BWF1 mice revealed the up-regulation of a numbers of genes coding for potentially inflammatory factors including Spp1 (Fig. 5a). The feature was reminiscent of a so-called senescence-associated secretory phenotype (SASP) [3], and indeed the expression of Cebpb, shown to be involved in the regulation of SASP, was also increased.

 

 

SA-T cells are senescence-associated T cells.

Edited by Turnbuckle, 06 March 2021 - 11:14 PM.

[userCK]

I did 3 cycles of Mito protocol and didn't feel any different - neither during fission nor fusion so I am assuming the damage to my mitos was not that high. So, I've now switched to SC Renewal protocol.

 

This is my first cycle and I wanted to ask others who have tried this, during part 2 (Senescent cell replacement), is it normal to feel weird/nauseous? Hard to describe the feeling but not a good feeling.

 

PS: I used higher dosage for most ingredients just-in-case the supplements were underdosed. Here is exactly what I used:

 

Part 1: Stem cell self-renewal

 

Time 0 —

Mango Butter 20g g (mixed in boiling water, should yield 10g of stearic acid)

 

Time 2:30 — (because Mango Butter digests faster)

 

Sulforaphane — 100 mg  150mg (5 x Broccomax activated sulforaphane)

Liposomal glutathione — 1 g  2grams, Core

SAM-e — 500 mg - 800mg (400mg x 2)

AAKG  — 2-3 g  7.2 grams (3.6g of each NOW and Sunday brand)

 

Time 3:00 —

C60 — 3 mg (in oil) 8mg (2 x 5ml of SES single serve packs, it says 80mg/100ml, so it should yield 8mg in 10ml)

Amino acids: Threonine — 2-3 g  6g, Lysine — 2 g (4g), Methionine — 1 g  2g, Leucine — 1 g 2g

(and repeated twice in a day)

 

Part 2: Senescent cell replacement (24 hours later or more)

Lysine — 2 g 4g, Methionine — 1g 2g

Nicotinamide — 2g 3g, Ribose — 2g 3g

Curcumin (liposomal or phytosomal) — 2g 3g

Azithromycin 500 mg (not planning to use it in second cycle, just this one)

Quercetin phytosome 250 mg 500mg

 

I'll re-do this cycle (but without Azithromycin) on March 18th. And then again on March 28th, And then again on April 8th. For a total of 4 cycles. Then on April 15th or so, I plan to collect saliva sample for TruMe DNA Age test to see if anything changed.  My last TruMe baseline is at 43 years old (my chronological age is 37).

 

 

Edited by userCK, 08 March 2021 - 02:26 PM.

[Turnbuckle]

So you doubled most everything up and you're asking is it normal to feel nauseous? Duh.

[JamesPaul]

2. Lithium Ortotate 50mg

 

 

While reading several dozen reviews of a lithium-containing supplement on amazon.com, I was struck by the huge range of positive and negative experiences.  A few people wrote something like "After a couple of decades of feeling dreadful prior to taking this lithium supplement, I started feeling normal and like myself again."  Others wrote "this made me feel worse".  And everything in between. It dawned on me that different people must have vastly different needs for and tolerances of lithium.  Perhaps some people don't assimilate it well from food, or excrete it readily, and others people may be designed in the opposite way.  50 mg lithium sounds way too high, at least to start with, unless you have some knowledge of your system and its needs and tolerances.

[aribadabar]

While reading several dozen reviews of a lithium-containing supplement on amazon.com, I was struck by the huge range of positive and negative experiences.  A few people wrote something like "After a couple of decades of feeling dreadful prior to taking this lithium supplement, I started feeling normal and like myself again."  Others wrote "this made me feel worse".  And everything in between. It dawned on me that different people must have vastly different needs for and tolerances of lithium.  Perhaps some people don't assimilate it well from food, or excrete it readily, and others people may be designed in the opposite way.  50 mg lithium sounds way too high, at least to start with, unless you have some knowledge of your system and its needs and tolerances.

 

I read it as 50 mg Lithium ororate which is about ~1.5mg of elemental lithium.

[userCK]

 

So you doubled most everything up and you're asking is it normal to feel nauseous? Duh.

 

 

Completed second cycle of the protocol today. No nausea this time.

One or two more cycles and then I'd do the TruMe test again. Excited!

 

 

While reading several dozen reviews of a lithium-containing supplement on amazon.com, I was struck by the huge range of positive and negative experiences.  A few people wrote something like "After a couple of decades of feeling dreadful prior to taking this lithium supplement, I started feeling normal and like myself again."  Others wrote "this made me feel worse".  And everything in between. It dawned on me that different people must have vastly different needs for and tolerances of lithium.  Perhaps some people don't assimilate it well from food, or excrete it readily, and others people may be designed in the opposite way.  50 mg lithium sounds way too high, at least to start with, unless you have some knowledge of your system and its needs and tolerances.

 

Normal prescription is closer to 1000mg a day. So, 50mg is quite low.

I read it as 50 mg Lithium ororate which is about ~1.5mg of elemental lithium.

True.

[Turnbuckle]

Update on my epigenetic age.

 

This is an update on my personal experiment with age reversal, with 2 more data points. The previous update was in post 1471. There was nothing special during the last two tests. Just a handful of stem cell treatments and a 2 week mito treatment that included AKG. As expected, neither the mito treatment nor the additional AKG had much additional effect on epigenetic age, which held steady at around -22 years, though I definitely benefited from the mito treatment energetically.

 

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 (+ cinnamon powder)

08/2020*.... -11.3 (+ cinnamaldehyde & eugenol)

 

10/2020*.... -22.6 (+ Arginine AKG, 2-3 grams)

10/2020**... -18.1 (+ Arginine AKG, 2-3 grams)

 

1/2021* ..... -20.5 (+ Arginine AKG, 2-3 grams -- very few from 10/2020 to 3/2021)

3/2021* ..... -22.4 (2 week daily fission/fusion mito treatment with AKG)

 

 

* TruMe test

** EpiAging test

[eighthman]

I am finding great success with this regimen.   I alternate with the regimen ( two days) and then use autophagy related supplements across 5 days - figuring that these may contradict each other and cycling is needed.

 

I did wonder what supplements might be tested next in the regimen.   There doesn't seem to be much info about trying to duplicate/imitate Yamanaka factors as a do it yourself pursuit.  If we were to witness 30 - 40 epigenetic years lost, some heads may explode !

[kurdishfella]

C60 helped me whistle. Couldnt do it otherwise. Nootropics help with that. I think deficient in neurotrophins causes problems with jaw function or somth.
Other than that I didnt feel like i Healed any better. maybe only helps unhealthy folks.

Edited by kurdishfella, 30 March 2021 - 03:37 PM.

[OlderThanThou2]

Does AKG demethylate the mtDNA? mtDNA seems to be methylated more at non-CpG sites. I believe the TET activated by AKG work on CpG sites.

 

Human mitochondrial DNA is extensively methylated in a non-CpG context | Nucleic Acids Research | Oxford Academic (oup.com)

 

[Turnbuckle]

Does AKG demethylate the mtDNA? mtDNA seems to be methylated more at non-CpG sites. I believe the TET activated by AKG work on CpG sites.

 

Human mitochondrial DNA is extensively methylated in a non-CpG context | Nucleic Acids Research | Oxford Academic (oup.com)

 

Appears so--

 

...it has become apparent that DNA methylation could take place in the mitochondrial genome, especially in the main non-coding region [14,15,16], which contains the regulatory regions OH, HSP1/2, and LSP. This is further supported by the finding that DNA methyltransferases, namely the DNMT enzymes, and DNA demethylation enzymes, the ten-eleven translocation methylcytosine dioxygenases (TET), have been found in mitochondria [17, 18]. The DNMT enzymes methylate cytosine into 5-methylcytosine (5mC), whereas the TET enzymes promote DNA demethylation by oxidizing 5mC to 5-hydroxymethylcytosine (5hmC) [17, 18]. This is also consistent with our previous finding that the DNA demethylation agents vitamin C (VitC), the activator of TET, and 5-Azacytidine (5Aza), the inhibitor of DNMT, were able to significantly reduce levels of mtDNA methylation [16]. 

https://www.ncbi.nlm...les/PMC6296150/

 

 

 

Edited by Turnbuckle, 31 March 2021 - 02:18 PM.

[OlderThanThou2]

Ok, thanks. It gives one more reason to take AKG. I wonder if epigenetic clocks would make sense for the mtDNA.

[Turnbuckle]

 I wonder if epigenetic clocks would make sense for the mtDNA.

 

 

Those presently available only look at nuclear DNA.

[OlderThanThou2]

I wonder which one would demethylate faster between the DNA and mtDNA and also if using the mito protocol would reduce the mitochondrial methylation.

[kurt9]

I am finding great success with this regimen.   I alternate with the regimen ( two days) and then use autophagy related supplements across 5 days - figuring that these may contradict each other and cycling is needed.

 

I did wonder what supplements might be tested next in the regimen.   There doesn't seem to be much info about trying to duplicate/imitate Yamanaka factors as a do it yourself pursuit.  If we were to witness 30 - 40 epigenetic years lost, some heads may explode !

 

There is a discussion thread in here on the cellular reprogramming with Yamanaka factors or other approaches. But there is still more research to be done before we can do this as a DIY protocol.

[kurt9]

Would this protocol help with lung conditions such as COPD and fibrosis?

[Turnbuckle]

Would this protocol help with lung conditions such as COPD and fibrosis?

 

 

Since mitochondrial dysfunction is believed to be involved with the pathogenesis of both, you should look to the mitochondrial protocol, not this one.

 

https://www.longecit...ndpost&p=903440

[kurt9]

Well, since I think mitochondrial dysfunction, even more than epigenetic issues, is the primary cause of aging and metabolic syndrome, what you may makes sense to me. Indeed, I think a lot of the epigenetic damage is itself caused by mitochondrial dysfunction. In any case, I plan on doing your new mitochondrial routine this summer (once I finish ALA chelation).

 

BTW, mitochondrial dysfunction may be the root of both. But the senolytics with stem-cell replacement may be useful in repairing existing damage once the underlying mitochondrial problems are fized. I do not have either of these two lung conditions, that I know of. But given my childhood history of asthma, lung health is something I take more seriously than most people I know. It is my concern about the masks (for social distancing) releasing friable particles and their affect on lung health that prompted my question in the first place. I do not want my shot at radical life extension to be cut short by preventable lung disease.

[Fafner55]

My epigenetic test results from TruMe

I followed the Turnbuckle protocol for the last 4 months and tested epigenetic age every 2 months with the following outcome.

 

Date         (Epigenetic age - Chronological age), years

12/17/2020.... -10.45      Baseline before epigenetic treatments

02/17/2021.... -10.32      Turnbuckle protocol every 10 days (6 times total)  with 120 mg gotu kola, During this period I did 1 fasting mimicking diet,

04/13/2021.... -12.58      Turnbuckle protocol every 10 days (6 times total).

 

My current chronological age is nearly 66. I am in good health.

 

The exact protocol I followed was

Time 0 —

• Stearic acid — 10 g (2 tsp glyceryl monostearate)

Time 3:00 hr —

• Sulforaphane — 100 mg
• Glutathione (NAC + Glycine + Vitamin C) — 1 g each
• Methyl folate, 400 ug   (consider changing to methyl donor SAM-e — 500 mg)
• Arginine-alpha-ketoglutarate (AAKG)  — 2 g

Time 3:30 hr —

1. C60 — 3 mg (1 tsp, in MCT oil, homemade)
2. Amino acids: 
   1. Threonine — 2 g
   2.  Lysine — 2 g
   3. Methionine — 1 g
   4. Leucine — 1 g

 

Other background -

Since 2016 I have cleared senescent cells 14 times with 120 mg dasatinib + other BCL2 inhibitors, most recently (11/2020) with 1600 mg fisetin + 1200 mg quercetin phytosome.

As for exercise, when the weather permits I take a 30 minute brisk walk per day.

 

While these results trend in the right direction, they do not show the dramatic -20 year reversal that Turnbuckle achieved nor a significant change from my baseline.

 

[dlewis1453]

 

Other background -

Since 2016 I have cleared senescent cells 14 times with 120 mg dasatinib + other BCL2 inhibitors, most recently (11/2020) with 1600 mg fisetin + 1200 mg quercetin phytosome.

As for exercise, when the weather permits I take a 30 minute brisk walk per day.

 

While these results trend in the right direction, they do not show the dramatic -20 year reversal that Turnbuckle achieved nor a significant change from my baseline.

 

Hi Fafner55, 

 

This is great! Thanks for sharing these very interesting results. Every additional data point helps. 

 

My first thought on why you did not achieve results as drastic as Turnbuckle is that you had already undertaken very aggressive clearance of senescent cells before beginning Turnbuckle's protocol. The fact that you had already achieved a ~10 year epigenetic age reduction from senescent cell clearance is already a great achievement. A reduction of epigenetic age of ~2 years in 4 months from Turnbuckle's protocol is still a good outcome in my opinion. 

 

Have you noticed any improvements to your health or appearance since beginning your earlier senescent cell clearance treatment or since beginning Turnbuckle's treatment?

[yz69]

Well, I had a wild ride on TB's stem cell protocol. Did a baseline test in 01/2020 with a trume age 47.5, now just received my latest result at 47.7.

I did the test every month. The lowest number I got was 38.8, highest was 48.6, most were around 45.

[dlewis1453]

Well, I had a wild ride on TB's stem cell protocol. Did a baseline test in 01/2020 with a trume age 47.5, now just received my latest result at 47.7.

I did the test every month. The lowest number I got was 38.8, highest was 48.6, most were around 45.

 

Thats very interesting yz69. Were you only taking Turnbuckle's protocol during this time? Or were you taking any telomerase activators as well? 

[yz69]

I was only doing TB’s protocol with daily extra vitamin d3. When I started the protocol, for the first few months, things were going pretty well, every month I saw a drop on the TruMe age until one day I bit a bone when eating a burger and cracked my tooth, the TruMe age went from 38.8 to 46.
After the tooth heals the TruMe age went down a bit but shot up to 48.6 in January 2021, can’t figure out what caused that. In February the number went down to 45, now the March number just came in at 47.7. It could be that the March test was done just 10 days after my 2nd COVID vaccine shot. I was feeling awful after that shot.

[Fafner55]

 

Have you noticed any improvements to your health or appearance since beginning your earlier senescent cell clearance treatment or since beginning Turnbuckle's treatment?

 

The short answer is no. My health and appearance seem the same since beginning of these rounds of the Turnbuckle Protocol.  Previously, an earlier variation of this protocol quickly healed a long-lasting knee injury of mine (Post #204), which lends support to the idea that stem cell proliferation is increased.

 

Clearing senescent cells made measurable changes. My erythrocyte sedimentation rate dropped from 5 to 2 and my CRP dropped from 1.75 to 0.3. I measure those markers nearly every year and they remain low. No doubt there are consequences to methylation patterns, but I don't know which genes Trume measures or the extent the SASP might modulate its estimations.

 

Anecdotally, my doctor commented that I have the lowest sed rate of any of his older patients, and my ophthalmologist volunteered that I have less sign of cataracts than her other patients my age. As for my appearance, my skin is typical of a 65 year old, maybe a bit better, but I do look my age.

[OlderThanThou2]

My epigenetic test results from TruMe

I followed the Turnbuckle protocol for the last 4 months and tested epigenetic age every 2 months with the following outcome.

 

Date         (Epigenetic age - Chronological age), years

12/17/2020.... -10.45      Baseline before epigenetic treatments

02/17/2021.... -10.32      Turnbuckle protocol every 10 days (6 times total)  with 120 mg gotu kola, During this period I did 1 fasting mimicking diet,

04/13/2021.... -12.58      Turnbuckle protocol every 10 days (6 times total).

 

My current chronological age is nearly 66. I am in good health.

 

 

 

 

How would you qualify your diet overall during your life?

 

Does it look rather like this one:

 

https://www.aging-us...icle/202913/pdf

 

on page 9427?

[Turnbuckle]

I was only doing TB’s protocol with daily extra vitamin d3. When I started the protocol, for the first few months, things were going pretty well, every month I saw a drop on the TruMe age until one day I bit a bone when eating a burger and cracked my tooth, the TruMe age went from 38.8 to 46.
After the tooth heals the TruMe age went down a bit but shot up to 48.6 in January 2021, can’t figure out what caused that. In February the number went down to 45, now the March number just came in at 47.7. It could be that the March test was done just 10 days after my 2nd COVID vaccine shot. I was feeling awful after that shot.

 

I had a strong reaction to the second Moderna shot -- flu like symptoms without fever for nearly a week -- so my next test will be able to say something about that. However I don't expect any reversal as I feel better than ever. This likely has to do with the mito protocol, but that by itself had little or no apparent effect on my epigenetic age. I don't understand why your numbers shot up, but I had that happen the first year. The increase was very dramatic and it took a long time to recover the lost ground. The bad actor was cycloastragenol, which I incorrectly thought would have minimal effect if I took it only with the protocol. When I stopped it, my epigenetic age began declining again. Lengthening telomeres blocks replacement, and the unused stem cells you created may be recycled by homeostatic mechanisms and thus wasted. Other substances that are said to lengthen telomeres include--

 

TA-65

Vitamin D

Centella asiatica, Astragalus, oleanolic acid (OA), and maslinic acid 

Marine omega-3 fatty acids

 

There are probably many more among supplements in common use that haven't been studied. The biggest problem will be with substances that lengthen the shortest telomeres, so the numbers may not be all that impressive as a telomerase enhancer, yet make it poison for this protocol.

Edited by Turnbuckle, 22 April 2021 - 05:42 PM.