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Manipulating mitochondrial dynamics

nad nad+ c60 mito fission fusion stearic acid mtdna methylene blue

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#1351 Arcanist

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Posted 14 February 2019 - 05:05 PM

It's also important to keep in mind that glucose-deprivations, as in low or no-carb, results in physiological insuline-resistance too by shifting Insulin-receptor transcription.

 

Subsisting on FFS is no good for mitos as a general rule, especially when they are PUFAs. I understand that Niacinamide also protects Mitos from high FFS influx/blocks it.

 

 

What's the consensus about Methylen Blue and mito-manipulation. It shifts the NAD+/NADH-ratio towards NAD+ without the need to consume Niacinamide. Maybe supplementing MB would not hamper Fusion while still providing the longevity benefits of a higher NAD+ to NADH ratio?


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#1352 Arcanist

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Posted 19 February 2019 - 09:16 AM

https://www.liebertp...9/rej.2018.2077

 

 

Abstract

Nicotinamide adenine dinucleotide (NAD+) is an essential pyridine nucleotide that serves as an electron carrier in cellular metabolism and plays a crucial role in the maintenance of balanced redox homeostasis. Quantification of NAD+:NADH and NADP+:NADPH ratios are pivotal to a wide variety of cellular processes, including intracellular secondary messenger signaling by CD38 glycohydrolases, DNA repair by poly(adenosine diphosphate ribose) polymerase (PARP), epigenetic regulation of gene expression by NAD-dependent histone deacetylase enzymes known as sirtuins, and regulation of the oxidative pentose phosphate pathway. We quantified changes in the NAD+ metabolome in plasma samples collected from consenting healthy human subjects across a wide age range (20–87 years) using liquid chromatography coupled to tandem mass spectrometry. Our data show a significant decline in the plasma levels of NAD+, NADP+, and other important metabolites such as nicotinic acid adenine dinucleotide (NAAD) with age. However, an age-related increase in the reduced form of NAD+ and NADP+—NADH and NADPH—and nicotinamide (NAM), N-methyl-nicotinamide (MeNAM), and the products of adenosine diphosphoribosylation, including adenosine diphosphate ribose (ADPR) was also reported. Whereas, plasma levels of nicotinic acid (NA), nicotinamide mononucleotide (NMN), and nicotinic acid mononucleotide (NAMN) showed no statistically significant changes across age groups. Taken together, our data cumulatively suggest that age-related impairments are associated with corresponding alterations in the extracellular plasma NAD+ metabolome. Our future research will seek to elucidate the role of modulating NAD+ metabolites in the treatment and prevention of age-related diseases.

Introduction

In the last decade, there has been growing interest in the role of redox active nucleotides in the metabolism.1 The significance of pyridine nucleotide coenzymes, such as nicotinamide adenine dinucleotide (NAD+) and its phosphorylated form NADP+, as main electron transfer molecules and substrates for over 700 oxidoreductase enzymes is undebated.2 We and others have previously demonstrated that disturbances in the redox balance, for example, following exposure to chronic oxidative stress, often represents an important component of the pathobiology of cell loss in cardiovascular and neurodegenerative diseases.3,4 Exogenous stressors, such as overfeeding, starvation, alcohol ingestion, or drug treatment can alter the intracellular redox status of these coenzymes.5

NAD+ represents one of the most important coenzymes in the hydride transfer reactions.6 NAD+ is the precursor of the pyridine nucleotide family, including NADH, NADP+, and NADPH, and is the end product of tryptophan metabolism via the kynurenine pathway.7 It has been well established that NAD+ is a substrate for major dehydrogenase enzymes involved in nutrient catabolism, including alcohol and lactate dehydrogenase reactions.8 As well, NADH, which is the reduced form of NAD+, preferentially provides electrons to power mitochondrial oxidative phosphorylation. Apart from its roles in fuel utilization, NAD+ also serves as an exclusive substrate for the nuclear repair enzymes poly(adenosine diphosphate [ADP] ribose) polymerases (PARP). PARPs are a family of enzymes that are activated by double- or single-stranded DNA breaks in DNA, and are thought to promote DNA repair by the ADP-ribosylation of histones and other nuclear proteins.9 NAD+ is also a substrate for the enzyme NAD+ glycohydrolases (CD38) that leads to the production of cyclic ADP-ribose, a calcium efflux effector.10 NAD+ has also been shown to be the sole substrate for a new class of NAD-dependent histone deacetylase (“HDAC”) enzymes known as sirtuins.11 Increasing histone acetylation is associated with age-related pathologies, whereas gene silencing by upregulation of sirtuins has been shown to extend lifespan in yeast and small organisms.12 HDACs are also being found to interact with a variety of nonhistone proteins and to thereby change their function, activity, and stability by post-translational modifications.

Accurate determination of the NAD+ metabolome is of major interest due to its potential association with cognitive decline, including AIDS dementia complex,13–15 cancer,16–18 aging, and a plethora of age-related disorders. Recently, nicotinic acid adenine dinucleotide (NAAD), an intermediate of NAD+ synthesis from nicotinic acid (NA) via the NAD+ salvage pathway, has been shown to increase following ingestion with niacin.19 This finding suggests that increased NAD+ anabolism by supplementation with NAD+ precursors not only increases the accumulation of by-products of NAD+ catabolism (such as ADP-ribose and N-methyl-nicotinamide [MeNAM]), but also stimulates retrograde synthesis of NAAD and nicotinic acid mononucleotide (NAMN). However, the mechanism responsible for this elusive biochemical reaction is yet to be identified.

Given the significance of the NAD+ metabolome in a multitude of biological processes, accurate quantification of its concentration and redox state in plasma and tissue is essential for better understanding of important biochemical processes, and determining the metabolic state of organisms in response to treatment with various compounds and disease states. We and others have shown that the NAD+:NADH ratio varies between 1 and 10 in catabolic tissue of “physiologically” aged female Wistar rats, and human subjects.3,4,20 As NAD+ also serves as an oxidative agent in some biochemical processes such as fatty acid oxidation, glycolysis, and citrate cycle, changes to the NAD+:NADH ratio may also represent an indicator of alterations in metabolic processes and several diseases including multiple sclerosis. In 2011, we were the first to prove that NAD+ is an essential factor in the aging process in major declining levels of catabolic tissue such as the brain, heart, lung, liver and kidney of rats, and in human pelvic tissue.3,4,20 Increased NAD+ anabolism has been shown to ameliorate mitochondrial dysfunction in a mechanism dependent on SIRT1, a nuclear sirtuin.

While it is thought that NAD+ is predominantly an intracellular nucleotide, emerging evidence suggests that extracellular NAD+ crosses the plasma membrane and replenishes intracellular NAD+.21 Intracellular NAD+ concentrations have been shown to range between 10 and 1000 μM, and are much higher than the levels reported in the extracellular space.22 This is because (1) NAD+ is released from cells at low amounts; (2) NAD+ catabolism is rapid leading to biologically active products; and (3) NAD+ directly interacts with cell surface receptors such as connexion 43 channels and several subtypes of purinergic P2 receptors.23

Therefore, accurate monitoring of the plasma NAD+ metabolome is necessary and may provide valuable information regarding the effect of various lifestyle and dietary factors, pharmacological and nutraceutical supplementation of NAD+ and/or its metabolites. Monitoring the plasma NAD+ metabolome levels will also allow drug candidates to be screened for a new type of potentially adverse effect—the depletion of NAD+ and/or other desirable metabolites. Moreover, the ratio (e.g. the NAD:NADH ratio) of oxidized and reduced forms of pyridine dinucleotides provides important information regarding redox metabolism disorders or alterations to cellular bioenergetics and may become important biomarkers for the early detection of pathological states.


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#1353 Arcanist

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Posted 19 February 2019 - 09:57 AM

So supplementig NAD+ precursors might not yield pronounced longevity or anti-age effects when you're already aged or in aging in general.

 

Inhibiting CD38 and taking Methylene Blue might be a better strategy for longevity effects of NAD/NADH-Balance modulation if not for mitos.


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#1354 ambivalent

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Posted 19 February 2019 - 10:21 PM

Turnbuckle, with any of your protocols have you noticed any produce increase in facial volume? Or any suggestions on how to regain some. 



#1355 jgkyker

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Posted 21 February 2019 - 04:28 AM

Turnbuckle, with any of your protocols have you noticed any produce increase in facial volume? Or any suggestions on how to regain some. 

 

I'm working on an elastin boosting protocol and have been considering creating a new thread to explain it. Enhancing elastin production may cause the skin to regain elasticity.

 

I don't have time to go into all the details right now. At some point down the road, I may be able to post more details. I probably won't post if it doesn't work, and I want to give it a month or so to see what happens. I think it is doing something... in only a few days. But I keep telling myself that is all in my head.


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#1356 jgkyker

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Posted 22 February 2019 - 01:11 AM

Turnbuckle, with any of your protocols have you noticed any produce increase in facial volume? Or any suggestions on how to regain some. 

 

Hey, I've been reading through this thread, here and there, and I remembered that Turnbuckle mentioned using a Derma-roller and a lysine solution he made. I have not tried this. Therefore, I cannot speak to it.

 

Here is the Google search with the relevant pages in this thread:

https://cse.google.c...dynamics/page-7



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#1357 Psy

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Posted 23 February 2019 - 02:31 AM

I'm working on an elastin boosting protocol and have been considering creating a new thread to explain it. Enhancing elastin production may cause the skin to regain elasticity.

 

 

Would be very nice. I am very interesting to push elastin too.


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#1358 QuestforLife

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Posted 24 February 2019 - 03:10 PM

Mitophagy without fission?

http://jcs.biologist...tent/124/8/1339

I'm keen to get the forum's thoughts on this paper.

#1359 Turnbuckle

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Posted 24 February 2019 - 04:11 PM

Mitophagy without fission?

http://jcs.biologist...tent/124/8/1339

I'm keen to get the forum's thoughts on this paper.

 

Doesn't apply to humans.



#1360 Phoebus

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Posted 24 February 2019 - 09:21 PM

I'm working on an elastin boosting protocol and have been considering creating a new thread to explain it. Enhancing elastin production may cause the skin to regain elasticity.

 

I don't have time to go into all the details right now. At some point down the road, I may be able to post more details. I probably won't post if it doesn't work, and I want to give it a month or so to see what happens. I think it is doing something... in only a few days. But I keep telling myself that is all in my head.

 

Please do make that thread, even a failed experiment is worth reporting as we all learn from it 

 

 


Edited by Phoebus, 24 February 2019 - 09:35 PM.

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#1361 jgkyker

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Posted 25 February 2019 - 04:45 AM

I'm working on an elastin boosting protocol and have been considering creating a new thread to explain it. Enhancing elastin production may cause the skin to regain elasticity.

 

I don't have time to go into all the details right now. At some point down the road, I may be able to post more details. I probably won't post if it doesn't work, and I want to give it a month or so to see what happens. I think it is doing something... in only a few days. But I keep telling myself that is all in my head.

 

Sorry this is off-topic, but due to the interest, I thought I would link the thread I created discussing this: https://www.longecit...otion-protocol/


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#1362 Arcanist

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Posted 28 February 2019 - 08:08 AM

Caution maybe is advised if supplementing with high doses of NAD+-precursors:

 

 

 

https://wistar.org/n...wlRYVB9rQAid9qs

 

PHILADELPHIA — (Feb. 18, 2019) — Researchers at The Wistar Institute have described a novel role of nicotinamide adenine dinucleotide (NAD+) metabolism in the ability of senescent cells to release tumor-promoting molecules. This study, published online in Nature Cell Biology, also cautions that NAD+-augmenting supplements, currently in development for their anti-aging effects as nutroceuticals, should be administered with precision given their potential pro-tumorigenic side effects.

Cellular senescence is a process in which cells irreversibly stop dividing and represents a potent tumor suppressive mechanism. At the same time, senescent cells also produce a variety of inflammatory soluble molecules that can promote tumor growth, known by the name of senescence-associated secretory phenotype (SASP).

The laboratory of Rugang Zhang, Ph.D., deputy director of The Wistar Institute Cancer Center, professor and co-program leader of the Gene Expression and Regulation Program, investigated the role of a family of proteins called HMGAs in cellular senescence and the SASP.

HMGAs are DNA-binding proteins that regulate gene expression. They are frequently overexpressed and associated with poor prognosis in many cancer types, yet they are known to promote senescence. This new research aims to clarify these mechanisms to unveil the dual role of the HMGA family of proteins in cancer.
 
Zhang and colleagues discovered that in cells undergoing senescence HMGAs increase the levels of NAMPT, a key enzyme in the production of nicotinamide adenine dinucleotide (NAD+), a cellular factor critical for metabolism and enzyme function. Importantly, increased NAD+ levels enhanced the SASP.

“Our data show that NAD+ levels influence the secretory activity of senescent cells in a way that may promote tumor progression,” said Zhang. “It is well documented that cellular NAD+ concentrations decrease during aging and the use of NAD+ supplementation is being studied as a new preventive opportunity for aging and age-associated disorders. Our results may have far-reaching implications on this field of investigation.” 

In fact, by manipulating the expression of HMGA1 and NAMPT, the researchers observed that increased NAD+ metabolism promotes cancer cell proliferation and progression in vivo in mouse models of pancreatic and ovarian cancers. 

These data point to a model whereby increased NAD+ metabolism resulting from higher HMGA1 and NAMPT expression promotes a higher SASP and enhances the inflammatory environment around the tumor, which in turn has a stimulatory effect on cancer growth.

“We found that HMGA1 provides a link between the two opposite sides of the coin in the process of senescence — growth arrest and the tumor promoting, proinflammatory secretory phenotype,” said Zhang. 

“Our data raise the possibility that targeting NAMPT may be an effective approach to suppress a proinflammatory, tumor promoting microenvironment in tumors that overexpress HMGA1 when treated with senescence-triggering cancer therapeutics such as chemotherapy and radiotherapy,” said Timothy Nacarelli, Ph.D., first author of the study and a postdoctoral researcher in the Zhang Lab. “NAMPT inhibitors are currently in clinical trials and thus readily available for these new applications.”


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#1363 jgkyker

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Posted 05 March 2019 - 07:49 PM

Caution maybe is advised if supplementing with high doses of NAD+-precursors:

 

 

 

https://wistar.org/n...wlRYVB9rQAid9qs

 

 

Here are my first thoughts on this. NAD+ is one of the key components for cellular respiration, which is how cells create their energy. So, if you have an immune system cell, boosting NAD+ could boost its function. Alternatively, if you have a tumor cell, boosting NAD+ could boost the tumor's capability to produce energy as well.

 

Elevated NAD+ levels in younger people theoretically do the same thing.

 

What am I missing? Perhaps, one concern is older people may have significantly more senescent cells to deal with on a day-to-day basis. (I have not researched this area yet. So, I really have no definitive understanding of this.) If older people are boosting NAD+, they are probably "feeding" their senescent cells, just as younger people do. However, the difference is that there are greater quantities, and perhaps, greater quantities translate into higher possibility for tumors.


Edited by jgkyker, 05 March 2019 - 07:54 PM.

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#1364 jgkyker

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Posted 06 March 2019 - 03:14 AM

NAD+ is one of the key components for cellular respiration, which is how cells create their energy. So, if you have an immune system cell, boosting NAD+ could boost its function. Alternatively, if you have a tumor cell, boosting NAD+ could boost the tumor's capability to produce energy as well.

 

Check it out, I am partially wrong, according to this study:

 

Biology of glucose metabolization in cancer cells

 

Cancer cells typically “switch” from “cellular respiration” to the very inefficient glycolysis for their ATP needs, a phenomenon described by Otto Warburg in 1924. The amazing thing is that tumors, which are highly energy demanding tissues, switch to a very inefficient energy producing pathway. They make up for this energy demand by going through glycolysis faster than necessary in normal cells.

 

 

Fascinating stuff.

 

However, glycolysis utilizes NAD+ and reduces it to NADH. So, boosting NAD+ theoretically provides more "ammo" for glycolysis, so-to-speak, and especially hungry tumor cells which, according to the above study, "[Go] through glycolysis faster than necessary in normal cells."

 

For those that are unfamiliar with glycolysis and its use of NAD+, check out this site at Penn State on glycolysis.

 

glycolysis_energy.jpg?version=1&modifica

 

My next step of action is to begin studying senolytics heavily... I'm already including Quercetin and cinnamon on fission promotion days... but I'm beginning to think I need to understand senolytics much more clearly before everything else...


Edited by jgkyker, 06 March 2019 - 03:31 AM.


#1365 Valijon

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Posted 06 March 2019 - 04:39 PM

Ive looked into increasing NAD+ through supplementation. I don't think it is doing nuch. To me at least, we are dealing with oral substances that dont have great bioavailability. Then theres the issue of half life. It seems too short to have much, if any, benefits. I would focus more on senolytics. We have known about Dasatanib for seceral years. We need more senolytics and harder hitting ones.

#1366 Turnbuckle

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Posted 15 March 2019 - 03:52 PM

I'm reposting this from post 977, as I left off some info that those who come across this from my profile page will not know. Like the purpose of this protocol, and how to take stearic acid.
 

 

Purpose: The average cell contains some one thousand mitochondria, each which contains several identical loops of mitochondrial DNA (mtDNA). With aging or with drug use (like statins), loops of mtDNA develop mutations more rapidly and cells become less efficient at removing them. Even one mutation is sufficient to shut down ATP production if this loop is alone in the mitochondria. Thus this protocol fissions all mitochondria to the minimum size containing one loop by increasing the NAD+/NADH ratio. At this maximum level of fission, cells can easily identify and remove those defective loops by the process of mitophagy. These are then replaced via biogenesis, whereby mitochondria are fused with stearic acid, and new mtDNA loops are produced using PQQ and the remaining mitochondrial mtDNA as templates. If you have a large portion of defective mtDNA*, many cycles of this 5-day protocol will be needed. You can use it once a week, or space it out as desired.
 
* The proportion of defective mtDNA in a cell can go up to 100%, in which case the damage cannot be reversed.

 

 

Day 1-2: Fission/mitophagy

 

Morning

Nicotinamide (NAD+) — 2g 

Ribose (NAD+) — 2g

Jiaogulan leaf (AMPK activator) — 1g

Apigenin (fission) — 100mg

Fisetin (Sirt1 activator) — 100mg

 

Evening

Jiaogulan leaf — 500mg

Fisetin — 100 mg

 

Day 3: Fission/mitophagy

 

Morning & Evening

Jiaogulan leaf  — 500mg

Fisetin — 100 mg

 

Day 4: Fusion/biogenesis

 

Morning

Stearic acid (fusion) — 10 g (cook into brownies and take 3 hours before other ingredients, see notes below)

Leucine (biogenesis) — 5g

PQQ (biogenesis) — 20mg

Hydroxytyrosol (biogenesis & antioxidant) — 25mg

Vitamin B complex (commercial mix)

 

Evening

Leucine — 5g (cook into 

PQQ — 20mg

Hydroxytyrosol — 25mg

 

Day 5: Fusion/biogenesis

 

Morning & evening

Leucine  — 5g

PQQ  — 20mg

Hydroxytyrosol  — 25mg

 

 

Notes:

 

Stearic acid — use either “food grade” or NF/USP grade, three hours before taking other ingredients. This has to be cooked to become bioavailable, and its digestion is slow even when cooked. It can be baked into brownies-- 

 

Directions: Using a box mix that calls for 2/3 cups of oil, I cut that back to 2 tablespoons and add 120 grams of stearic acid flakes, leaving the rest of the recipe unchanged. Then I mix at room temp using a power mixer, baking according to directions, dividing it 3x4 and freezing most of it for later use. They can be eaten without thawing.

 

 

Alternatively you can substitute 5 grams of glycerol monosterate (mixing it into hot chocolate or a hot food like oatmeal) and take it at the same time as the other ingredients. It is far more rapidly digested and thus may rapidly push up blood pressure. This can be controlled with BP medications and/or 5-10g taurine and a gram or two of oleopein.

 

Hydroxytyrosol — if unavailable where you live, leave it off.

 

If you believe you have a great deal of damage, you might start off with only one day of fission/mitophagy. 


Edited by Turnbuckle, 15 March 2019 - 04:25 PM.

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#1367 Dteshome85

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Posted 15 March 2019 - 05:54 PM

Thanks for sharing this Mr.Turnbuckle

#1368 stephen_b

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Posted 16 March 2019 - 07:47 AM

I'm happy to see glycerol monosterate as an alternative to stearic acid in the protocol. Thanks for sharing.



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#1369 zorba990

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Posted 20 March 2019 - 05:22 PM

1g agmatine 3g citrulline most certainly lowers bp for me. I only take on workout days now as the effect seems to last days and be somewhat cumulative.





Also tagged with one or more of these keywords: nad, nad+, c60, mito, fission, fusion, stearic acid, mtdna, methylene blue

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