2284 replies to this topic

[Turnbuckle]

 

I've had real problems with what I believe to be an arthritic knee for over a year - I assume it had just worsened with time but now I see that it has coincided with starting PT (and NR) - my knee has been much better the last few months and I've greatly reduced PT by coincidence; however, just lately I've started to increase it again and have moderate discomfort. Obviously I'm not highly confident it is PT yet, the peak of my knee inflammation coincided with several weeks of very challenging walking but also very high doses of NR+PT, so it was easy to overlook supplements as a cause, but it should be easy to be ascertain at some point (I've also experienced no other joint pain). I've been ramping up NR again and have noticed the sole of my feet pain I first experienced when taking NR, but perhaps it was the PT. Incidentally, I've been taking NR intermittently with N+R for several weeks but  not noticed the joint or sole pain til lately, though NR dosing has increased. So perhaps we could see if there is correlation with PT and the joint pains experienced.

 

 

A number of people on the NR personal experience thread have reported joint pain--and appear to associate it with NR.

 

When I look at the pathways involved, nampt is the enzyme that converts nicotinamide to NMN.

 

==================================

 

If you are successful in upping your NAD+, you will generate NADP, and the NADP will be converted into a number of subsidiary chemicals--including nicotinamide (the so-called 'salvage pathway').[See attached diagram.}

 

This could easily lead to higher nampt production in at least some people, and greater inflammation. Too much NAD+ on a continual basis might have consequences, even without pterostilberine.

 

One of the nice things about Turnbuckle's protocol is that it is cyclic, rather than simply swamping the body with the same supplement day-after-day.

 

 

 

You make a good point. The first time I tried fission/exercise I did it 4 days straight and subsequently felt beat up for a week--which included knee and ankle pain. As I wasn't using resveratrol at the time, nampt could have been the source of that. So a nampt inhibitor might be even more useful than fusion.

[able]

I've seen more than a few references that NAMPT is NOT pro-inflammatory as first thought. Or depends on the situation.  For instance, this one:

 

"Nicotinamide Phosphoribosyltransferase (NAMPT) was initially believed to be a cytokine, and given the name pre-B enhancing colony factor, and later an insulin-mimetic hormone called Visfatin (205-207). The first claim has not been supported and remains inconclusive, while evidence supporting Visfatin has been retracted. It is now known as one of a few enzymes involved in the NAD+-salvage pathway. "

 

http://ir.lib.uwo.ca...491&context=etd

 

 

Not conclusive.  Just saying, I wouldn't try to inhibit NAMPT myself.

[Andey]

There is an interesting study published yesterday in Nature (I stumble on it in fightaging.org).

It's too technical for me but one of the conclusions is that shifting dynamics to more fission, less fusion promotes lifespan (at least in flies). Another thing that was mentioned I don't know about is that its well-observed tendency to increase the average size of mitochondria with age.

https://www.nature.c...467-017-00525-4

Edited by Andey, 07 September 2017 - 08:06 AM.

[Turnbuckle]

There is an interesting study published yesterday in Nature (I stumble on it in fightaging.org).

It's too technical for me but one of the conclusions is that shifting dynamics to more fission, less fusion promotes lifespan (at least in flies). Another thing that was mentioned I don't know about is that its well-observed tendency to increase the average size of mitochondria with age.

https://www.nature.c...467-017-00525-4

 

 

This thread is a descendant of another one in which getting rid of zombie mitochondria was a major goal. See post 24, and the first link in that post.

[ighbal]

Just a 5 cts comment:

 

Looks like this paper : https://www.ncbi.nlm...les/PMC5028859/

 

says that Omega 3 promotes fusion. 

[Turnbuckle]

Seems that I linked to the wrong page two posts above. The post 24 referring to zombie mitochondria can be found here--

 

http://www.longecity...ndpost&p=772985

 

And the paper describing zombie mitochondria can be found here--

 

http://www.ncbi.nlm....les/PMC4392818/

 

When normal mitophagic organelle elimination is suppressed by Parkin insufficiency, abnormal undead or zombie mitochondria accumulate and (as zombies will do) contaminate the normal mitochondrial population by fusing with normal organelles. Mitochondrial fusion that is ordinarily protective, therefore, becomes the mechanism for a general contagion of mitochondrial dysfunction. Interrupting mitochondrial fusion prevents contamination of functionally normal mitochondria by virulent zombie mitochondria, sequestering abnormal mitochondria that can then be removed by alternate, albeit almost certainly less efficient, elimination pathways.

 

Inhibiting mitochondrial fusion to contain mitochondrial contagion induced by mitophagic dysfunction may have applications in human diseases caused by defective mitochondrial quality control, most notably hereditary Parkinson disease induced by PINK1 and Parkin mutations.

 

 

Zombies are not removed by normal mito QC, so the idea is to augment those QC processes until they do. For instance, by stimulating genes that are known to augment fission and reduce fusion, and by greatly increasing the NAD+/NADH ratio that also stimulates fission and sets the QC process running. This should not be done continuously, but only intermittently.

Edited by Turnbuckle, 10 September 2017 - 01:19 PM.

[Nate-2004]

Just a 5 cts comment:

 

Looks like this paper : https://www.ncbi.nlm...les/PMC5028859/

 

says that Omega 3 promotes fusion. 

 

This is interesting. I normally don't skip a single day of DHA/EPA just because it's been shown that it significantly slows the  age related loss of brain volume over time.

 

I wonder what the case is for EVOO? I typically take a TBSP of DeCarlo EVOO every day (not C60).

[Nate-2004]

This video discusses mitochondrial fusion and fission in relation to brown fat and insulin. Mitochondrial dynamics discussed at about 10 mins in.

 

Edited by Nate-2004, 11 September 2017 - 07:41 PM.

[Nate-2004]

Based on the statements in this video which I have watched twice now, it sounds like keto for fusion and high carb for fission? He said insulin stimulates ceramides which induces fusion?

[ceridwen]

Very interesting. High carb makes me very ill. So what does that imply?

[aconita]

Possibly for you high carbs means high gluten, even if it hasn't to be necessarily so.

[xEva]

Based on the statements in this video which I have watched twice now, it sounds like keto for fusion and high carb for fission? He said insulin stimulates ceramides which induces fusion?

 

no, he said that ceramide accumulation in muscle induces mitochondrial fission. He said that sustained state of mitochondrial fission results in increased oxidative stress, reduced ATP production and changes the "overall metabolic profile". ~13:50+

 

Then he says he does not know why ketones cause mitochondrial uncoupling (and thus "waste energy" when it is needed most) and calls this a "philosophical question", but there is a simple answer to this. In short, he forgets to look at the bigger picture and count the net ATP that was spent by the cell on useful work:

 

Mitochondrial uncoupling is associated not only with diminished ATP production, because some of the effort went up in heat, but, more importantly, it leads to fewer ROS being produced -- and ROS are damaging to the cell. He forgets that that damage must be dealt with, consequently, which will also require ATP. So if he would count the ATP spent for the "useful work" by taking all ATP produced minus the ATP that went for repairs of the damage induced by ROS, he'd see that, even though uncoupling results in less ATP being produced, it makes the cell more efficient overall, because it also decreases oxidative stress and the need to deal with its consequences.  capisce?

 

Like many PhDs, he is somewhat nearsighted and focused on a narrow pathway and its immediate results. He needs to zoom out a bit and see a bigger picture. 

Edited by xEva, 13 September 2017 - 08:16 AM.

[Nate-2004]

 

Based on the statements in this video which I have watched twice now, it sounds like keto for fusion and high carb for fission? He said insulin stimulates ceramides which induces fusion?

 

no, he said that ceramide accumulation in muscle induces mitochondrial fission. He said that sustained state of mitochondrial fission results in increased oxidative stress, reduced ATP production and changes the "overall metabolic profile". ~13:50+

 

Then he says he does not know why ketones cause mitochondrial uncoupling (and thus "waste energy" when it is needed most) and calls this a "philosophical question", but there is a simple answer to this. In short, he forgets to look at the bigger picture and count the net ATP that was spent by the cell on useful work:

 

Mitochondrial uncoupling is associated not only with diminished ATP production, because some of the effort went up in heat, but, more importantly, it leads to fewer ROS being produced -- and ROS are damaging to the cell. He forgets that that damage must be dealt with, consequently, which will also require ATP. So if he would count the ATP spent for the "useful work" by taking all ATP produced minus the ATP that went for repairs of the damage induced by ROS, he'd see that, even though uncoupling results in less ATP being produced, it makes the cell more efficient overall, because it also decreases oxidative stress and the need to deal with its consequences.  capisce?

 

Like many PhDs, he is somewhat nearsighted and focused on a narrow pathway and its immediate results. He needs to zoom out a bit and see a bigger picture. 

 

 

I got that, but I'm trying to figure out what conclusions to draw from what he said, not what he actually said. How does ceramide accumulate in the muscle? Insulin from what I'm gathering. Thus why I concluded carbs on fission days.

 

Even with that explanation you gave, which I'm probably going to have to read a couple of times to understand, I'm still really confused about diet relative to fission or fusion.

[Andey]

Very interesting. High carb makes me very ill. So what does that imply?

 

Occam`s razor - check your BG levels 1 and 2 hours after high carbohydrate meal

 

 

 He forgets that that damage must be dealt with, consequently, which will also require ATP. So if he would count the ATP spent for the "useful work" by taking all ATP produced minus the ATP that went for repairs of the damage induced by ROS, he'd see that, even though uncoupling results in less ATP being produced, it makes the cell more efficient overall, because it also decreases oxidative stress and the need to deal with its consequences.  capisce?

 

Like many PhDs, he is somewhat nearsighted and focused on a narrow pathway and its immediate results. He needs to zoom out a bit and see a bigger picture. 

 

 

Looks to me like a wild speculation, any references to support this? 

 

 

Edited by Andey, 13 September 2017 - 05:16 PM.

[xEva]

I got that, but I'm trying to figure out what conclusions to draw from what he said, not what he actually said. How does ceramide accumulate in the muscle? Insulin from what I'm gathering. Thus why I concluded carbs on fission days.
 
Even with that explanation you gave, which I'm probably going to have to read a couple of times to understand, I'm still really confused about diet relative to fission or fusion.

He said that they induce ceramide biogenesis in rats by giving them an inflammatory agent (LPS or TNF-alpha -?),  and that overaccumulation of ceramide  in turn will lead to insulin resistance. Then he says essentially that 'the stick is of two ends' and that sustained insulin resistance, without inflammation, will also lead to ceramide accumulation. He makes a distinction between triglycerides and ceramide, both of which, according to him, are induced by insulin, though he calls overaccumulation of ceramide toxic and that of triglycerides benign. He says that the distinction between an athlete and a diabetic is that the first has triglycerides stored in muscle and the second, ceramide.

 

So, what conclusion can you draw?   That insulin resistance is bad -? ..and that you gotta eat less and exercise more  lol -- hardly a revelation.

 

 

Re uncoupling, it's a question of amount or dosage. The first well-known uncoupler was DNP. Depending on the dose, it can be a poison (still used as insecticide or dewormer in some farm animals) or "a dieting aid" for man. Taken internally, it induces a pleasant warmth sensation in the belly -- too much of it and it will turn to burning and can cause necrosis in the GI tract, which may result in death (and as a dieting aid it was abolished).

 

So it all depends on how much uncoupling a substance causes. Too much -- and no ATP is produced whatsoever and the cell dies. Just a little bit is good though (metformin is also a mild mitochondrial uncoupler -- just google for refs). There is no set point, it all depends on needs and circumstances.

 

Interesting that just before a "natural death", just 2-3 days before (like in some cancer patients, for example), people start experiencing internal heat and seek cold by pulling off blankets and asking to open windows -- that's because their mitochondria deteriorate, with membranes turning 'holey'. This was noticed by nurses long ago, allowing them to predict when a patient is about to die.

 

On the other hand, if you start practicing cold water plunging, you will experience a very, very pleasant sensation of a heat wave rolling through your body (just after you get out of water). That too is mitochondrial uncoupling. You can also experience it on a dry fast -- suddenly, a very pleasant wave of warmth washes over you. In these two cases it has a long-lasting effect of elevated mood and energy level. 

[xEva]

Looks to me like a wild speculation, any references to support this?

Plenty:  just google UCPs ROS

 

ex:

Mitochondrial ROS metabolism: modulation by uncoupling proteins,  2001

Uncoupling proteins and the control of mitochondrial reactive oxygen species production, 2011

[Andey]

 

Looks to me like a wild speculation, any references to support this?

Plenty:  just google UCPs ROS

 

ex:

Mitochondrial ROS metabolism: modulation by uncoupling proteins,  2001

Uncoupling proteins and the control of mitochondrial reactive oxygen species production, 2011

 

 

Your claim was that repairing damage from ROS takes a substantial part of ATP production and the net effect of uncoupling gives equal or greater ATP in cells disposal. It's where you said that Mr Bikman is a short sighted person )

 

P.S. Sorry for off topic, it's just gone too far in a weird direction recently. 

Edited by Andey, 13 September 2017 - 06:27 PM.

[Turnbuckle]

 

 

 

P.S. Sorry for off topic, it's just gone too far in a weird direction recently. 

 

 

 

I agree. It does not move the thread forward in a useful way.

[Nate-2004]

He said that they induce ceramide biogenesis in rats by giving them an inflammatory agent (LPS or TNF-alpha -?),  and that overaccumulation of ceramide  in turn will lead to insulin resistance. Then he says essentially that 'the stick is of two ends' and that sustained insulin resistance, without inflammation, will also lead to ceramide accumulation. He makes a distinction between triglycerides and ceramide, both of which, according to him, are induced by insulin, though he calls overaccumulation of ceramide toxic and that of triglycerides benign. He says that the distinction between an athlete and a diabetic is that the first has triglycerides stored in muscle and the second, ceramide.

 

 

That makes more sense now.  However, why are ceramides bad? Loss of ceramides in skin is part of what leads to wrinkles. There's even facial creams and pills you can take to add them back. I assume he just means in muscle tissue?

 

So, what conclusion can you draw?   That insulin resistance is bad -? ..and that you gotta eat less and exercise more  lol -- hardly a revelation.

 

Well no he said that doesn't work and I tend to agree, eat less exercise more isn't the solution because it really doesn't work, not for losing any weight and for people trying CR it's a miserable experience for most people. Hell, even fasting is apparently easier than CR. Insulin resistance is bad though and I think to keep that down I'm going to have to try to aim for either doing keto in cycles (which is what they did in the recent rat study) or just avoid carbs as much as I can. For now I'm going to do a month of keto starting with a fast and see how I feel at the end of it all.

 

So it all depends on how much uncoupling a substance causes. Too much -- and no ATP is produced whatsoever and the cell dies. Just a little bit is good though (metformin is also a mild mitochondrial uncoupler -- just google for refs). There is no set point, it all depends on needs and circumstances.

 

I'm taking berberine in 300mg doses 3 times a day, I wonder if this is the same thing. I wonder how it affects fission/fusion. I started taking it to see how it affects essential tremor, which is why I'm giving keto a solid month to try and see what happens.

 

On the other hand, if you start practicing cold water plunging, you will experience a very, very pleasant sensation of a heat wave rolling through your body (just after you get out of water). That too is mitochondrial uncoupling. You can also experience it on a dry fast -- suddenly, a very pleasant wave of warmth washes over you. In these two cases it has a long-lasting effect of elevated mood and energy level.

 

I thought of doing this, I don't have the ice for it, but I wonder how long you have to be in there for that to happen. I should start a gym with heat and cold stress facilities.  

 

Edited by Nate-2004, 13 September 2017 - 09:00 PM.

[Nate-2004]

 

 

 

 

P.S. Sorry for off topic, it's just gone too far in a weird direction recently. 

 

 

 

I agree. It does not move the thread forward in a useful way.

 

 

What's so off topic about it? This thread is about manipulating mitochondrial dynamics and I don't think we've gotten off topic from that at all, because if diet is relevant, inflammation, insulin, etc, and not just timing supplements with exercise, then that's important I think no?

[aconita]

Simplicity is the ultimate sophistication - Leonardo da Vinci

[Andey]

 

 

 

What's so off topic about it? This thread is about manipulating mitochondrial dynamics and I don't think we've gotten off topic from that at all, because if diet is relevant, inflammation, insulin, etc, and not just timing supplements with exercise, then that's important I think no?

 

 

   I think the whole protocol idea is that it gives an overwhelming signal to induce fission. Any dietary intervention should be on verge of noise ratio to it. I believe studies on effects of elevated NAD+ levels kinda support it because there is not much variability in results but apparently different subjects had different diets.

  One could argue that prolonged nutritional ketosis ( the subject of the podcast with Benjamin Bikman) is strong enough intervention by itself and could interfere with the protocol. That's where you could test it and report your observations here. That's the gist of the thread.  

   I am now in a nutritional ketosis for some quite prolonged time and fission part of protocol feels exactly the same as on a high carbohydrate diet. I don't do fusion part exactly, I just eat cocoa liquor and have much longer intervals between cycles as it should do the same thing.

Edited by Andey, 14 September 2017 - 06:27 AM.

[Turnbuckle]

Two-path NAD+ experimental protocol

Rationale: Tryptophan provides a second path to NAD+

 

Before bed—Fission/mitophagy combination

nicotinamide — 1.5g

tryptophan — 1g

ribose — 5g

Life Extension AMPK activator — 500mg

Fisetin (Sirtuin activator) — 100mg

 

Morning—More ribose if necessary

 

Before lunch—Biogenesis

PQQ — 10mg

Acacia catechu extract — 580 mg (one cap)

Antioxidants (presently using 2g C, 25mg HT, and 1g Setria glutathione)

 

Results: I’ve done this every day for a week with no adverse effects. I sleep better (due to the serotonin from the tryptophan) and wake up with amazing muscle tone, as if I’ve been to the gym in my sleep. I go to the gym in the morning and find that I have none of the “exercise like a girl” weakening, as it appears that the strong fissioning caused by NAD+ is already gone. Thus I see no need to use fusion promoters.

 

I plan to continue this at least for another week.

 

Note: Diet presently high protein/low carb, and has been for some months

 

[BigLabRat]

Very interesting. Both nicotinamide and tryptophan are sleep inducers for me, so this would probably knock me for a loop. (That's not a bad thing.)

 

One question: How do you know of more ribose is needed? Didn't follow that bit.

[Turnbuckle]

Very interesting. Both nicotinamide and tryptophan are sleep inducers for me, so this would probably knock me for a loop. (That's not a bad thing.)

 

One question: How do you know of more ribose is needed? Didn't follow that bit.

 

 

Yes, they are sleep inducers, and that's why I am taking them before bed. The half life or ribose is several times less than nicotinamide or tryptophan. I know  I need more when I get the same slightly odd feeling I get when taking nicotinamide without ribose--which is why I wasn't a fan of nicotinamide years back. But after taking more ribose, the feeling goes away. 

[albedo]

 

Two-path NAD+ experimental protocol

Rationale: Tryptophan provides a second path to NAD+ ........

Thank you for your very informative posts. I am not an expert on NAD+ but can you confirm if my understanding of Tryptophan as second path is the one depicted in the attached picture? I understand NAD+ increases via the "amidated" path (e.g. NR, NMN, NAM), "deamidated" path (e.g. NA) or "de novo synthesis" from tryptophan:

 

 NAD.PNG   75.73KB   0 downloads

 

Katsyuba E, Auwerx J. Modulating NAD(+) metabolism, from bench to bedside. EMBO J. 2017

http://onlinelibrary....201797135/full

 

[BigLabRat]

Yes, tryptophan comes into the process through the same "arm" as niacin (nicotinic acid).

 

Tryptophan's pathway (the 'de novo' pathway) merges with niacin's at nicotinic acid mononucleotide (NaMN).The tryptophan route to NaMN is pretty complicated, and has to convert tryptophan to quinolinic acid first--but it gets there!

 

By contrast, NR is the same "arm" as nicotinamide (the 'salvage pathway'--so-called because nicotinamide can either be ingested, or recycled from byproducts of NAD use).

 

If the goal is to maximize NAD+, it makes sense to use both arms.

 

 

[tunt01]

Here is a pretty good graphic covering the various pathways.  It comes from the paper MikeDC posted in another thread, in part, authored by Mark Mattson.

 

 

 

Fang EF, e. (2017). NAD(+) in Aging: Molecular Mechanisms and Translational Implications. - PubMed - NCBI. Ncbi.nlm.nih.gov. Retrieved 16 September 2017, from https://www.ncbi.nlm...from=nad aging

[albedo]

Thank you BigLabRat and Prophets for the explications and the chart.

 

I was trying to learn well Turnbuckle's post and also better understand an often cited paper on NAD boosting via NR which also interestingly says: "...We also report that nicotinic acid adenine dinucleotide (NAAD), which was not thought to be en route for the conversion of NR to NAD⁺, is formed from NR and discover that the rise in NAAD is a highly sensitive biomarker of effective NAD⁺ repletion..."

 

Trammell SA, Schmidt MS, Weidemann BJ, et al. Nicotinamide riboside is uniquely and orally bioavailable in mice and humans. Nat Commun. 2016;7:12948.

https://www.nature.c...les/ncomms12948

[Turnbuckle]

Yes, tryptophan comes into the process through the same "arm" as niacin (nicotinic acid).

 

Tryptophan's pathway (the 'de novo' pathway) merges with niacin's at nicotinic acid mononucleotide (NaMN).The tryptophan route to NaMN is pretty complicated, and has to convert tryptophan to quinolinic acid first--but it gets there!

 

By contrast, NR is the same "arm" as nicotinamide (the 'salvage pathway'--so-called because nicotinamide can either be ingested, or recycled from byproducts of NAD use).

 

If the goal is to maximize NAD+, it makes sense to use both arms.

 

 

I've also tried the combinations of--

 

1. nicotinamide + nicotinic acid + R

2. nicotinamide + tryptophan + R, and

3. nicotinamide + tryptophan + nicotinic acid + R

 

No. 3 was a total wipe out. I tried it during the day and could't get up the energy to do anything.

No. 1 was good for daytime use, unless one is sensitive to the niacin flush.

No. 2 is very good for nighttime use.

 

I haven't tried combinations without nicotinamide.