2255 replies to this topic

[Heisok]

Warner, I hope that you do not mind the partial quote, and yes this should probably go in the personal experience thread, but it is based on the interview.

 

I will not directly quote Dr. Brenner, but my understanding was that he said there was a peak about 8 hours after dosing. If one wanted a second peak, they could dose again for another peak 8 hours later.  My experience is similar to what Warner had. My dosing was in the A.M, but then a second dose around 5 pm. I started to not be able to fall back to sleep when I would wake up at 2 or 3 AM. Wide awake and thinking about stuff. I mentioned this sleep issue elsewhere, and my understanding of what Nate-2004 said is that they have had  insomnia, but they do not have problems on N.R. About 45 days ago, I completely discontinued N.R. to see if it was effecting my sleep. The result was that when I awoke at night, I could fall back to sleep, even if I had something on my mind.

 

Based on the video discussing the 8 hour peak, (thanks Bryan), and the very well thought out single dosing rationale which Nate-2004 has come up with, I restarted N.R. I take my full dose first thing in the morning on an empty stomach. I have been able to fall back to sleep if I wake up at night, in spite of having things on my mind.

 

 

"125 mg at dinner.  From the Brenner interview, he noted that NAD+ peaks in humans sometime during the afternoon, and then again in the middle of the night, and that one could optionally take a dose of NR at dinner to augment the nightly NAD+ peak (i.e., so both NAD+ peaks would be augmented, although he wasn't himself doing that).  Now, for various reasons (positional apnea control, etc.), my wife and I have an alarm set at 2:30 AM to get us up to pee, and we rarely have any trouble getting back to sleep after that.  However, after upping the NR dose to 375 mg/d, we're both having trouble getting back to sleep at that time.  There is no pain, etc.  We're just wide awake, thinking about stuff, taking much longer to fall back to sleep.  Some others have also noted that taking NR before bed makes sleeping more difficult.  I wouldn't have made much of this, but the fact that it's happening to both of us, under similar conditions, after upping the NR dose, with others reporting similar effects, makes me think its the NR."

Edited by Heisok, 30 October 2016 - 10:03 PM.

[warner]

Hi Heisok.  If you want a proper-looking partial quote, just press the "Quote" button and then edit its contents, as I did here when partially quoting your post:

I will not directly quote Dr. Brenner, but my understanding was that he said there was a peak about 8 hours after dosing. If one wanted a second peak, they could dose again for another peak 8 hours later.  My experience is similar to what Warner had. My dosing was in the A.M, but then a second dose around 5 pm. I started to not be able to fall back to sleep when I would wake up at 2 or 3 AM. Wide awake and thinking about stuff. I mentioned this sleep issue elsewhere, and my understanding of what Nate-2004 said is that they have had  insomnia, but they do not have problems on N.R. About 45 days ago, I completely discontinued N.R. to see if it was effecting my sleep. The result was that when I awoke at night, I could fall back to sleep, even if I had something on my mind.

 

Based on the video discussing the 8 hour peak, (thanks Bryan), and the very well thought out single dosing rationale which Nate-2004 has come up with, I restarted N.R. I take my full dose first thing in the morning on an empty stomach. I have been able to fall back to sleep if I wake up at night, in spite of having things on my mind.

Yup, I think Bryan also reported having trouble sleeping if NR is taken too late in the day. (?)

 

As I recall, Brenner was talking about 2 different types of peaks:  the 8 hr NR peak after each dosing, versus the 2 daily natural peaks in human NAD+ (during day and in middle of night).  By taking NR first thing in the morning, then you can time your NR peak to roughly match the natural NAD+ peak, and by taking NR near dinner time, then you can time your next NR peak to roughly match the natural nighttime NAD+ peak (although he doesn't do that).  However, for many of us, doing the latter appears to cause some insomnia (and where my alarm at 2:30 AM just happened to be near the NR peak from the dinner dose).  [I don't actually understand why its best to match the NR with the NAD+ peaks, but I'll take Brenner's word for it at this point.  ]

 

Also, as I recall, in the interview, Brenner makes it pretty clear that you don't need to take NR on an empty stomach (he does do that, but then immediately eats his breakfast).  That's because the NR is quite water-soluble and stable, and not affected by fat, etc., from food, and will still be well-absorbed in the presence of food.  (There's apparently several companies looking into adding NR to various foods.)

Edited by warner, 31 October 2016 - 02:37 AM.

[warner]

I didn't see this already listed and it seemed pertinent...

 

Metabolic Response to NAD Depletion across Cell Lines Is Highly Variable.

https://www.ncbi.nlm...les/PMC5053472/

 

From the discussion:

 

Here we examine the underlying consequences of NAD depletion with the NAMPT inhibitor GNE-617 on global metabolism in four different NSCLC [non-small cell lung carcinoma] cell lines. Even though there are published reports of metabolic profiling following NAMPT inhibition, there is no systematic study that compares the effects of NAD depletion across sensitive and resistant cell lines. We show that the effects of NAD depletion are surprisingly divergent across NSCLC cell lines and we propose that some of these differences may be driven by genetic variability...

 

We hypothesize that the underlying metabolic rate of a cell, or tumor, may be an important contributor to sensitivity, as a cell line with a slow doubling time may have a low metabolic turnover and thus be more resistant to NAD depletion. Additionally, our data demonstrates that the underlying profile of each cell line can modulate the metabolic response to NAMPT inhibition. This raises the possibility that inhibitors targeting different metabolic pathways may be effectively combined with a NAMPT inhibitor in some cell lines; however, these combinations may have to be tailored to match the underlying genetic and metabolic dependency of a particular cell...

 

In conclusion, our data reveal that depletion of a single key central metabolite such as NAD can have wide effects on overall cellular metabolism and that each cell line has a unique way of dealing with metabolic stress that can be dictated by its genetic profile.

 

I guess this suggests, in reverse (i.e., for NAD augmentation), that the effect of NR on cancer cells may be equally complex, dependent on the genetic variability between cancer types and subtypes. 

[Bryan_S]

 

Yup, I think Bryan also reported having trouble sleeping if NR is taken too late in the day. (?)

 

 

Sleeping well regardless but I could understand if someone else felt more energetic who wasn't accustomed to the alertness it could bring. I've got this exercise machine called a Versaclimber and it kicks my @$$. So I sleep pretty well.

[Bryan_S]

I guess this suggests, in reverse (i.e., for NAD augmentation), that the effect of NR on cancer cells may be equally complex, dependent on the genetic variability between cancer types and subtypes. 

 

 

We don't have enough data on this situation. In the liver study in mice it was beneficial. However if someone has cancer and wishes to start a NAD repletion regiment this should be discussed with their oncologist PERIOD.

[tunt01]

However, after upping the NR dose to 375 mg/d, we're both having trouble getting back to sleep at that time.  There is no pain, etc.  We're just wide awake, thinking about stuff, taking much longer to fall back to sleep.  Some others have also noted that taking NR before bed makes sleeping more difficult. 

 

 

 

SIRT1 is NAD+ dependent for its activity.  SIRT1 interacts with CLOCK::BMAL1 complex and participates with regulation of circadian rhythm.

 

 

Imai, S. (2010). “Clocks” in the NAD World: NAD as a metabolic oscillator for the regulation of metabolism and aging. Biochimica Et Biophysica Acta (BBA) - Proteins And Proteomics, 1804(8), 1584-1590. doi:10.1016/j.bbapap.2009.10.024  LINK

 

My understanding is that:

 

• CLOCK can act as an acetyltransferase and acetylates its partner BMAL1 (heterodimer construct).  SIRT1 inhibits this activity.  
• SIRT1 also degrades PER2 which builds up throughout the day and is ultimately part of the process that leads to a negative feedback process regulating circadian rhythm.

Availability of NAD+ is what is driving this metabolic, circadian rhythm process.  Others (Sassone-Corsi) have shown that SIRT1 levels are stable and it is oscillation of NAD+ that mediates this circadian rhythm regulation.

 

My (rather obvious) takeaway is that you want NAD+ (food, NR/NMN, etc.) during the waking hours and to avoid food during nighttime.  I found this metabolic pathway seems to explain why certain guidelines for dealing with jet lag suggest fasting and then immediately waking up and eating in the morning of whatever location one finds themself in.  Therefore, one shouldn't take NR or massive amounts of NAD+ supplementation/food late in the day.  It will disturb your circadian rhythm.

 

Anecdotally, several months ago I began eating only during daylight and taking one 125 mg NR in the first part of the day (~10 AM).  I quit eating by as early as 4 or 5 PM.  I've found an improvement in my circadian rhythm and a reduced frequency of awakenings at night.

 

There is a lot of work in this area done by Paolo Sassone-Corsi.  One of his reviews is on my todolist

 

Sassone-Corsi, P. (2012). Minireview: NAD+, a Circadian Metabolite with an Epigenetic Twist*. Endocrinology, 153(1), 1-5. doi:10.1210/en.2011-1535 [LINK]

 

By activating SIRT1, NAD+ conjoins two feedback loops necessary for cross talk between the circadian clock and metabolite production. The levels of the metabolite NAD+ were shown to oscillate in a circadian manner in various cell types, suggesting that it could determine the cyclic function of NAD+-dependent enzymes, such as SIRT1. It has been shown that the salvage pathway is critical to regulate intracellular NAD+ levels. After the conversion of NAM into NAM mononucleotide (NMN) by NAMPT, NMN is further modified into NAD+ by the NAM mononucleotide adenylyl transferases (Nmnat-1, -2, and -3). Although NAM inhibits SIRT1 activity, NAD+-activated SIRT1 feeds back into the NAD+ salvage pathway by directly regulating Nampt gene expression in a CLOCK:BMAL1-dependent manner (28). By this mechanism, NAD+ conjoins the two feedback loops, contributing to the fine tuning necessary for achieving energy balance.

 

 

Edited by prophets, 31 October 2016 - 11:58 PM.

[warner]

Thanks prophets for the excellent post about NAD and circadian rhythm.  Makes more sense that NAD levels are related to food consumption, rising during day, and falling at night.  And that augmenting them at night via NR might disrupt circadian rhythm.  (I'm following up on your ref.s, but it will take awhile to integrate it all.)

 

Well, first night without dinner NR went well for both of us - no problem getting back to sleep.  After a week of this may consider going back to 375 mg, but taking the last 125 mg no later than mid-afternoon (so last peak NR is not later than dinner digestion peak).

Edited by warner, 01 November 2016 - 02:10 AM.

[Gingerbread Man]

Thanks prophets for the excellent post about NAD and circadian rhythm.  Makes more sense that NAD levels are related to food consumption, rising during day, and falling at night.  And that augmenting them at night via NR might disrupt circadian rhythm.  (I'm following up on your ref.s, but it will take awhile to integrate it all.)

 

Well, first night without dinner NR went well for both of us - no problem getting back to sleep.  After a week of this may consider going back to 375 mg, but taking the last 125 mg no later than mid-afternoon (so last peak NR is not later than dinner digestion peak).

 

Very interesting on the circadian rhythm. It also seems to show how genetics and other differences in people change how NR reacts with them. For me, one of the first BIG changes NR made for me was how much it helps me sleep (from the very first night of taking it). I take it just before bed every night and I sleep better than I have in decades. I have been taking it for several months.

[stefan_001]

 

Thanks prophets for the excellent post about NAD and circadian rhythm.  Makes more sense that NAD levels are related to food consumption, rising during day, and falling at night.  And that augmenting them at night via NR might disrupt circadian rhythm.  (I'm following up on your ref.s, but it will take awhile to integrate it all.)

 

Well, first night without dinner NR went well for both of us - no problem getting back to sleep.  After a week of this may consider going back to 375 mg, but taking the last 125 mg no later than mid-afternoon (so last peak NR is not later than dinner digestion peak).

 

Very interesting on the circadian rhythm. It also seems to show how genetics and other differences in people change how NR reacts with them. For me, one of the first BIG changes NR made for me was how much it helps me sleep (from the very first night of taking it). I take it just before bed every night and I sleep better than I have in decades. I have been taking it for several months.

 

I take NR in morning and evening and also sleep well.

Edited by stefan_001, 01 November 2016 - 12:53 PM.

[Iporuru]

From Wikipedia: https://en.wikipedia...osyltransferase

 

"NAMPT is downregulated by an increase of miR-34a in obesity via a 3'UTR functional binding site of NAMPT mRNA resulting in a reduction of NAD(+) and decreased SIRT1 activity.^[4]"

 

From Neuroprotective action of lithium in disorders of the central nervous system: https://www.ncbi.nlm...MC3172812/#R66 

 

"In the rat hippocampus and in primary cultures of hippocampal neurons, chronic treatment with lithium or valproate decreases levels of several microRNAs, including miR-34a^[66],

(...)

It is further interesting to note that the protein level of Bcl-2 in the brains of a mouse model of AD is inversely correlated with the expression of miR-34a^[65], a microRNA that has recently emerged as a common lithium and valproate target^[66]. These findings suggest a novel mechanism for lithium's protective effects against AD in which the downregulation of miR-34a indirectly upregulates Bcl-2."

 

Edited by Iporuru, 01 November 2016 - 01:59 PM.

[Gingerbread Man]

 

 

Thanks prophets for the excellent post about NAD and circadian rhythm.  Makes more sense that NAD levels are related to food consumption, rising during day, and falling at night.  And that augmenting them at night via NR might disrupt circadian rhythm.  (I'm following up on your ref.s, but it will take awhile to integrate it all.)

 

Well, first night without dinner NR went well for both of us - no problem getting back to sleep.  After a week of this may consider going back to 375 mg, but taking the last 125 mg no later than mid-afternoon (so last peak NR is not later than dinner digestion peak).

 

Very interesting on the circadian rhythm. It also seems to show how genetics and other differences in people change how NR reacts with them. For me, one of the first BIG changes NR made for me was how much it helps me sleep (from the very first night of taking it). I take it just before bed every night and I sleep better than I have in decades. I have been taking it for several months.

 

I take NR in morning and evening and also sleep well.

 

Thanks Stefan, I am not ONLY taking at night before bed, I take it morning, mid day and just before bed (125mg per). So I don't spur confusion of thinking I only take it before bed (because others may not be aware of my dosages/usage). I was illustrating how my use of NR has impacted my sleep, opposite of what was being shown by some others.

[Nate-2004]

I tried taking 375mg before bed last night and I slept so hard that I overslept for work. I'm insomnia prone too so you would think it'd have caused insomnia for me but just the opposite.

[warner]

 

 

Thanks prophets for the excellent post about NAD and circadian rhythm.  Makes more sense that NAD levels are related to food consumption, rising during day, and falling at night.  And that augmenting them at night via NR might disrupt circadian rhythm.  (I'm following up on your ref.s, but it will take awhile to integrate it all.)

 

Well, first night without dinner NR went well for both of us - no problem getting back to sleep.  After a week of this may consider going back to 375 mg, but taking the last 125 mg no later than mid-afternoon (so last peak NR is not later than dinner digestion peak).

 

Very interesting on the circadian rhythm. It also seems to show how genetics and other differences in people change how NR reacts with them. For me, one of the first BIG changes NR made for me was how much it helps me sleep (from the very first night of taking it). I take it just before bed every night and I sleep better than I have in decades. I have been taking it for several months.

 

I take NR in morning and evening and also sleep well.

Okay, so a question that keeps bugging me is why one would only want to augment NAD+ during the daytime?  iow, if the point is that its average value declines with age, then it seems one would want to augment NAD+ during the full 24 hours, as long as this did not disrupt sleep, etc.  In fact, augmenting NAD+ when it is already near its daily peak may be the least helpful strategy, since the lower lows at night may be the biggest problem as we age.

 

This issue is exemplified by the current paper I'm reading:

 

https://www.ncbi.nlm...pubmed/23274583

 

Exp Eye Res. 2013 Mar;108:76-83. doi: 10.1016/j.exer.2012.12.007. Epub 2012 Dec 26.

NAD(+) maintenance attenuates light induced photoreceptor degeneration.

Bai S1, Sheline CT.

Abstract

Light-induced retinal damage (LD) occurs after surgery or sun exposure. We previously showed that zinc (Zn(2+)) accumulated in photoreceptors and RPE cells after LD but prior to cell death, and pyruvate or nicotinamide attenuated the resultant death perhaps by restoring nicotinamide adenine dinucleotide (NAD(+)) levels. We first examined the levels of NAD(+) and the efficacy of pyruvate or nicotinamide in oxidative toxicities using primary retinal cultures. We next manipulated NAD(+) levels in vivo and tested the affect on LD to photoreceptors and RPE. NAD(+) levels cycle with a 24-h rhythm in mammals, which is affected by the feeding schedule. Therefore, we tested the affect of increasing NAD(+) levels on LD by giving nicotinamide, inverting the feeding schedule, or using transgenic mice which overexpress cytoplasmic nicotinamide mononucleotide adenyl-transferase-1 (cytNMNAT1), an NAD(+) synthetic enzyme. Zn(2+) accumulation was also assessed in culture and in retinal sections. Retinas of light damaged animals were examined by OCT and plastic sectioning, and retinal NAD(+) levels were measured. Day fed, or nicotinamide treated rats showed less NAD(+) loss, and LD compared to night fed rats or untreated rats without changing the Zn(2+) staining pattern. CytNMNAT1 showed less Zn(2+) staining, NAD(+) loss, and cell death after LD. In conclusion, intense light, Zn(2+) and oxidative toxicities caused an increase in Zn(2+), NAD(+) loss, and cell death which were attenuated by NAD(+) restoration. Therefore, NAD(+) levels play a protective role in LD-induced death of photoreceptors and RPE cells.

 

So the gist of that is that raising NAD+ in various ways (Nam, NMNAT1, inverted feed schedule) protects retinal cells (photoreceptors and RPE) from light-induced damage.  And we can speculate that giving the rats some NR would have had the same positive result.  Further, giving the rats NR in the daytime (our nighttime), would have been particularly helpful.  The researchers mention AMD as an example of an eye disease that might be affected in this way.  In my own case, I have some glaucoma and a PED (pigment epithelial detatchment) that might be improved by higher levels of NAD+.  And of course we would all like to avoid AMD (age-related macular degeneration).

 

The hypothesis then might be that many of the problems associated with falling NAD+ with age may first begin with the lower lows of NAD+ that ageing adults experience at night (while sleeping), and that addressing these lows may be more important than trying to match NR dosage with daytime NAD+ levels.  Or, put another way, perhaps one should focus on taking enough NR during the day to ensure that enough of the augmented NAD+ is still available at night to offset the lower lows, but not so much as to disrupt sleep.

[tunt01]

Okay, so a question that keeps bugging me is why one would only want to augment NAD+ during the daytime?  iow, if the point is that its average value declines with age, then it seems one would want to augment NAD+ during the full 24 hours, as long as this did not disrupt sleep, etc.  In fact, augmenting NAD+ when it is already near its daily peak may be the least helpful strategy, since the lower lows at night may be the biggest problem as we age.

 

 

I don't have data support this, but my thought was that it is important to maintain a strong circadian rhythm and that as mammals age, the clock behavior (peripheral and suprachiasmatic) deteriorates somehat with age.  I think one of the open questions (for me at least) is if SIRT1 activity is dependent upon levels of NAD+, do you want constant availability of NAD+ and constant SIRT1 activity?  What is the downside and is there some kind of cancer risk here?

 

Of the many things that SIRT1 deacetylates, one of them is p53 an important checkpoint for cancer and cell cycle arrest.  Knowing that NAD+ oscillates over the course of a day, It seems risky to me to give the middle finger to mother nature and our epigenetic machinery and just ramp up NAD+ to high levels in perpetuity.  SIRT1 can be pro-oncogenic.

 

 

 

 

This issue is exemplified by the current paper I'm reading:

 

https://www.ncbi.nlm...pubmed/23274583

 

Exp Eye Res. 2013 Mar;108:76-83. doi: 10.1016/j.exer.2012.12.007. Epub 2012 Dec 26.

NAD(+) maintenance attenuates light induced photoreceptor degeneration.

Bai S1, Sheline CT.

Abstract

Light-induced retinal damage (LD) occurs after surgery or sun exposure. We previously showed that zinc (Zn(2+)) accumulated in photoreceptors and RPE cells after LD but prior to cell death, and pyruvate or nicotinamide attenuated the resultant death perhaps by restoring nicotinamide adenine dinucleotide (NAD(+)) levels. We first examined the levels of NAD(+) and the efficacy of pyruvate or nicotinamide in oxidative toxicities using primary retinal cultures. We next manipulated NAD(+) levels in vivo and tested the affect on LD to photoreceptors and RPE. NAD(+) levels cycle with a 24-h rhythm in mammals, which is affected by the feeding schedule. Therefore, we tested the affect of increasing NAD(+) levels on LD by giving nicotinamide, inverting the feeding schedule, or using transgenic mice which overexpress cytoplasmic nicotinamide mononucleotide adenyl-transferase-1 (cytNMNAT1), an NAD(+) synthetic enzyme. Zn(2+) accumulation was also assessed in culture and in retinal sections. Retinas of light damaged animals were examined by OCT and plastic sectioning, and retinal NAD(+) levels were measured. Day fed, or nicotinamide treated rats showed less NAD(+) loss, and LD compared to night fed rats or untreated rats without changing the Zn(2+) staining pattern. CytNMNAT1 showed less Zn(2+) staining, NAD(+) loss, and cell death after LD. In conclusion, intense light, Zn(2+) and oxidative toxicities caused an increase in Zn(2+), NAD(+) loss, and cell death which were attenuated by NAD(+) restoration. Therefore, NAD(+) levels play a protective role in LD-induced death of photoreceptors and RPE cells.

 

So the gist of that is that raising NAD+ in various ways (Nam, NMNAT1, inverted feed schedule) protects retinal cells (photoreceptors and RPE) from light-induced damage.  And we can speculate that giving the rats some NR would have had the same positive result.  Further, giving the rats NR in the daytime (our nighttime), would have been particularly helpful.  The researchers mention AMD as an example of an eye disease that might be affected in this way.  In my own case, I have some glaucoma and a PED (pigment epithelial detatchment) that might be improved by higher levels of NAD+.  And of course we would all like to avoid AMD (age-related macular degeneration).

 

The hypothesis then might be that many of the problems associated with falling NAD+ with age may first begin with the lower lows of NAD+ that ageing adults experience at night (while sleeping), and that addressing these lows may be more important than trying to match NR dosage with daytime NAD+ levels.  Or, put another way, perhaps one should focus on taking enough NR during the day to ensure that enough of the augmented NAD+ is still available at night to offset the lower lows, but not so much as to disrupt sleep.

 

 

IDK.  Maybe the LD cells were helped by NAD+ because they were already damaged.  Kind of like saying that obesogenic/T2D animals are helped by resveratrol, but resveratrol may not necessarily be of use to healthy individuals.  I'm not really sure, but just a comment for thought.

 

 

It may be that NAD+ through the salvage pathway is best upregulated during periods away from food and my original thought had it backwards.  In that, niacinamide inhibits SIRT1 in a postprandial fashion and that is the kind NAD+ production that should occur duing the day while, the salvage pathway is more active away from meals?  I don't know, I'm just shooting in the dark here.  But I'd really like to sit down and get a firmer grasp on this matter.

 

In my gut, I think the consumption NR and NAD+ circadian profile should mimic some kind of intermittent fasting pattern during the day in concert with sleep at night.  I'm not 100% clear on what the appropriate pattern is.  Maybe Brenner or Sassone-Corsi would have a cogent thought.

[Heisok]

Thanks to all for describing their positive experiences for sleep. I think that one member had severe pain which hurt sleep, and the N.R. seemed to help that a large amount. Overall, I feel that pain related to working out or working for hours digging in my yard was greatly reduced. That result started even on the 125 mg level which I took for about 18 months, before trying a higher dose. That enabled me to completely stop all NSAID style pain relievers (Tylenol or Advil) Interesting is that the pain moderating effect continued during the approximately 45 days which I stopped taking N.R. completely. Maybe, I improved my baseline NAD+ levels. I do not know if Osteoarthritis is hereditary, but other members of my family have problems with it.

 

As I said before, a morning dose, and then 5 pm dose did not work for me. I will not try that combination again. My physiology as it currently is is not compatible. If I decide to try split doses again, I will try splitting in the A.M., and then at bedtime. That test would be pretty far down the line, as this single dose in the morning regimen only started on 10/25/2016. Way too short of a test so far.

 

Nate was that your only dose for the day? Do you plan to test this bedtime dose for a longer time? One day is tough to project, especially if it was the only dose for the day. I am focusing on you due to the history of insomnia.

 

I tried taking 375mg before bed last night and I slept so hard that I overslept for work. I'm insomnia prone too so you would think it'd have caused insomnia for me but just the opposite.

 

 

Edited by Heisok, 01 November 2016 - 04:50 PM.

[Gingerbread Man]

 

Thanks to all for describing their positive experiences for sleep. I think that one member had severe pain which hurt sleep, and the N.R. seemed to help that a large amount. 

 

I tried taking 375mg before bed last night and I slept so hard that I overslept for work. I'm insomnia prone too so you would think it'd have caused insomnia for me but just the opposite.

 

That may have been me. I have had chronic back pain for a long time. It prevented me from getting any reasonable sleep. NR helped me sleep the first night I tried it but the back pain didn't decrease until about 2 weeks on it (with some added Ubiquinol in there too). I would say I still have a good 75% back pain reduction from the combination but it is easy to overdo it because of the decrease of pain and and up sore and hurting for a few days because of it.

[Harkijn]

 

Okay, so a question that keeps bugging me is why one would only want to augment NAD+ during the daytime?  iow, if the point is that its average value declines with age, then it seems one would want to augment NAD+ during the full 24 hours, as long as this did not disrupt sleep, etc.  In fact, augmenting NAD+ when it is already near its daily peak may be the least helpful strategy, since the lower lows at night may be the biggest problem as we age.

 

 

I don't have data support this, but my thought was that it is important to maintain a strong circadian rhythm and that as mammals age, the clock behavior (peripheral and suprachiasmatic) deteriorates somehat with age.  I think one of the open questions (for me at least) is if SIRT1 activity is dependent upon levels of NAD+, do you want constant availability of NAD+ and constant SIRT1 activity?  What is the downside and is there some kind of cancer risk here?

 

Of the many things that SIRT1 deacetylates, one of them is p53 an important checkpoint for cancer and cell cycle arrest.  Knowing that NAD+ oscillates over the course of a day, It seems risky to me to give the middle finger to mother nature and our epigenetic machinery and just ramp up NAD+ to high levels in perpetuity.  SIRT1 can be pro-oncogenic.

 

 

 

 

This issue is exemplified by the current paper I'm reading:

 

https://www.ncbi.nlm...pubmed/23274583

 

Exp Eye Res. 2013 Mar;108:76-83. doi: 10.1016/j.exer.2012.12.007. Epub 2012 Dec 26.

NAD(+) maintenance attenuates light induced photoreceptor degeneration.

Bai S1, Sheline CT.

Abstract

Light-induced retinal damage (LD) occurs after surgery or sun exposure. We previously showed that zinc (Zn(2+)) accumulated in photoreceptors and RPE cells after LD but prior to cell death, and pyruvate or nicotinamide attenuated the resultant death perhaps by restoring nicotinamide adenine dinucleotide (NAD(+)) levels. We first examined the levels of NAD(+) and the efficacy of pyruvate or nicotinamide in oxidative toxicities using primary retinal cultures. We next manipulated NAD(+) levels in vivo and tested the affect on LD to photoreceptors and RPE. NAD(+) levels cycle with a 24-h rhythm in mammals, which is affected by the feeding schedule. Therefore, we tested the affect of increasing NAD(+) levels on LD by giving nicotinamide, inverting the feeding schedule, or using transgenic mice which overexpress cytoplasmic nicotinamide mononucleotide adenyl-transferase-1 (cytNMNAT1), an NAD(+) synthetic enzyme. Zn(2+) accumulation was also assessed in culture and in retinal sections. Retinas of light damaged animals were examined by OCT and plastic sectioning, and retinal NAD(+) levels were measured. Day fed, or nicotinamide treated rats showed less NAD(+) loss, and LD compared to night fed rats or untreated rats without changing the Zn(2+) staining pattern. CytNMNAT1 showed less Zn(2+) staining, NAD(+) loss, and cell death after LD. In conclusion, intense light, Zn(2+) and oxidative toxicities caused an increase in Zn(2+), NAD(+) loss, and cell death which were attenuated by NAD(+) restoration. Therefore, NAD(+) levels play a protective role in LD-induced death of photoreceptors and RPE cells.

 

So the gist of that is that raising NAD+ in various ways (Nam, NMNAT1, inverted feed schedule) protects retinal cells (photoreceptors and RPE) from light-induced damage.  And we can speculate that giving the rats some NR would have had the same positive result.  Further, giving the rats NR in the daytime (our nighttime), would have been particularly helpful.  The researchers mention AMD as an example of an eye disease that might be affected in this way.  In my own case, I have some glaucoma and a PED (pigment epithelial detatchment) that might be improved by higher levels of NAD+.  And of course we would all like to avoid AMD (age-related macular degeneration).

 

The hypothesis then might be that many of the problems associated with falling NAD+ with age may first begin with the lower lows of NAD+ that ageing adults experience at night (while sleeping), and that addressing these lows may be more important than trying to match NR dosage with daytime NAD+ levels.  Or, put another way, perhaps one should focus on taking enough NR during the day to ensure that enough of the augmented NAD+ is still available at night to offset the lower lows, but not so much as to disrupt sleep.

 

 

IDK.  Maybe the LD cells were helped by NAD+ because they were already damaged.  Kind of like saying that obesogenic/T2D animals are helped by resveratrol, but resveratrol may not necessarily be of use to healthy individuals.  I'm not really sure, but just a comment for thought.

 

 

It may be that NAD+ through the salvage pathway is best upregulated during periods away from food and my original thought had it backwards.  In that, niacinamide inhibits SIRT1 in a postprandial fashion and that is the kind NAD+ production that should occur duing the day while, the salvage pathway is more active away from meals?  I don't know, I'm just shooting in the dark here.  But I'd really like to sit down and get a firmer grasp on this matter.

 

In my gut, I think the consumption NR and NAD+ circadian profile should mimic some kind of intermittent fasting pattern during the day in concert with sleep at night.  I'm not 100% clear on what the appropriate pattern is.  Maybe Brenner or Sassone-Corsi would have a cogent thought.

 

As I posted in the  Experiences thread I have been taking NR for quite some time now with very positive efffects. A contradictory development was that I developed cataracts and now I read about these rats I consider to switch(partly) to nighttime supplementation. (Also in that thread: for a short time I took NR before bed without any problems, but it seemed counterintuitive so I left off.)

[warner]

As I posted in the  Experiences thread I have been taking NR for quite some time now with very positive efffects. A contradictory development was that I developed cataracts and now I read about these rats I consider to switch(partly) to nighttime supplementation. (Also in that thread: for a short time I took NR before bed without any problems, but it seemed counterintuitive so I left off.)

It may be though that your daytime NR dose is already bleeding into your nighttime and giving your NAD+ levels a substantial lift compared to what may have been their relatively lower lows due to ageing.  So even if it is true that those NAD+ lower lows need lifting, it isn't clear how to best go about this without screwing up circadian rhythm, increasing cancer risk, etc.  I think that I'll probably split the difference and add a mid-afternoon 125 mg dose of NR (+ the 250 mg in morning) so that the NR peaks near the time of dinner meal digestion, with that raised NAD+ level bleeding into the night, but hopefully without disrupting circadian rhythm, etc.

Edited by warner, 01 November 2016 - 07:54 PM.

[Iporuru]

Don't know if this research has been posted: http://www.ergo-log....e-riboside.html

[warner]

As I began looking further into NAD-related circadian research, all the mechanistic diagrams showing the interaction of various factors weren't telling me squat about actual relative levels of these factors (like NAD+) during the diurnal cycle.  So here are a few circadian charts illustrating such diurnal changes.  Note that the charts that start with "0" hours on the X-axis are referring to "0" circadian time, which is the time of waking and first light exposure in diurnal animals (i.e., a person-specific time, typically 6-8 AM).

 

A few things:  Peak NAD+ is near dinner time (if you believe their model), so if you wanted to maintain that with NR supplementation, then you wouldn't want to take NR any later than mid-afternoon.  NAD+ falls during the night, reaching a minimum sometime before waking.  My hypothesis was that this minimum NAD+ is lowered with ageing, creating a vulnerability (a lower low of NAD+) in sum subsystems, such as retinal cells exposed to periods of low oxygen, glucose, blood flow, etc. (think glaucoma, AMD, etc.).  However, to counter that lower low, it may be sufficient to raise the lower high during the daytime, thereby preserving the diurnal cycle.  Interestingly, as NAD+ falls, melatonin rises, but that too is reduced with ageing (I already take a melatonin supplement to counter that).

 

Back to research!

 

from "Predicted Role of NAD Utilization in the Control of Circadian Rhythms during DNA Damage Response"

https://www.ncbi.nlm...pubmed/26020938

 

 circ NAD+ et al.PNG   1.18MB   2 downloads

 

from "Circadian rhythms and breast cancer"

http://caloriesprope...-breast-cancer/

 

 circ other stuff.jpg   125.86KB   2 downloads

 

from "Melatonin sensitizes the system"

http://caloriesprope...zes-the-system/

 

 circ melatonin.png   79.2KB   2 downloads

 

[tunt01]

Thanks so much for this post, Warner. I think I came across that 2nd to last graph when I was looking at this. You can see per1/2 rise dramatically during the beginning of sleep as a negative feedback regulator.

It seems these findings are in harmony with my original thought : NAD+ consumption in the early/mid day.

Edited by prophets, 02 November 2016 - 10:33 PM.

[normalizing]

hi i just read NR can regenerate liver and kidney cells, whats the best dose for this? as i read up to 500mg is well tolerated with most effect achieved around 300mg and above ok thank you

[warner]

I think I came across that 2nd to last graph when I was looking at this. You can see per1/2 rise dramatically during the beginning of sleep as a negative feedback regulator.  It seems these findings are in harmony with my original thought : NAD+ consumption in the early/mid day.

Yeah, the change in overall dynamics near dinner time is dramatic.  Also surprised to see that NAD+ is so circadian-dependent, although the plot/model shown obviously doesn't include any typical perturbations.  For purposes of timing the bulk of NR dosing, these plots are helpful I guess, but I still am left wondering about the overnight "lower lows" issue, and how best to deal with that for specific diseases.  Or perhaps a lot of that will need to be treated locally, as in NR eye drops for AMD (or whatever), perhaps applied only at night, etc.  Lots of work yet to be done.

 

(wrt modelling stuff, I recall that in the GRAS approval document for NR that came out late 2015, they noted that they had no measurements of NR (vs. its metabolites) to report since they hadn't figured out yet how to measure NR reliably!  Brenner et al. has since solved that problem, but it illustrates just how far we are from seeing exactly what's going on - we have a hard time just measuring much of the stuff we are trying to model.)

Edited by warner, 03 November 2016 - 03:57 AM.

[stefan_001]

 

I think I came across that 2nd to last graph when I was looking at this. You can see per1/2 rise dramatically during the beginning of sleep as a negative feedback regulator.  It seems these findings are in harmony with my original thought : NAD+ consumption in the early/mid day.

Yeah, the change in overall dynamics near dinner time is dramatic.  Also surprised to see that NAD+ is so circadian-dependent, although the plot/model shown obviously doesn't include any typical perturbations.  For purposes of timing the bulk of NR dosing, these plots are helpful I guess, but I still am left wondering about the overnight "lower lows" issue, and how best to deal with that for specific diseases.  Or perhaps a lot of that will need to be treated locally, as in NR eye drops for AMD (or whatever), perhaps applied only at night, etc.  Lots of work yet to be done.

 

(wrt modelling stuff, I recall that in the GRAS approval document for NR that came out late 2015, they noted that they had no measurements of NR (vs. its metabolites) to report since they hadn't figured out yet how to measure NR reliably!  Brenner et al. has since solved that problem, but it illustrates just how far we are from seeing exactly what's going on - we have a hard time just measuring much of the stuff we are trying to model.)

 

 

My reasoning for this one is quite different, a cell in trouble will be in trouble whether you sleep or not. The circadian rhythm is a system behavior like many other functions of the body and without doubt has a purpose. The question is probably more does NR supplementation mess up the rhythm and as result something else get negatively impacted. But for the individual cell health I dont see we need to maintain it.

 

(ps the rating of you post is not mine)

[warner]

My reasoning for this one is quite different, a cell in trouble will be in trouble whether you sleep or not. The circadian rhythm is a system behavior like many other functions of the body and without doubt has a purpose. The question is probably more does NR supplementation mess up the rhythm and as result something else get negatively impacted. But for the individual cell health I dont see we need to maintain it.

 

(ps the rating of you post is not mine)

 

When I was younger, that's the way I thought about it too.  But I learned the hard way that older mammals have problems that younger mammals don't, and that nighttime in particular can be dangerous for them.  In my case, calorie restriction (and some occasional apnea) got me in trouble with glaucoma and a PED (pigment epithelial detachment), both of which are accelerated by low oxygen, low glucose (relative hypoglycemia), low blood pressure (i.e., ischemia, hypoxia) which were occurring at night while sleeping.  So the paper I cited a few posts back about NR being helpful in protecting retinal cells from such conditions was of special interest.  But I think in general nighttime can be dangerous in older mammals, in that the drop in nutrient levels and blood pressure, combined with generally lowered NAD+ (and melatonin, etc.) with ageing, can put at risk any subsystem (eyes, heart, brain, etc.) that happens to be particularly susceptible to such conditions.  Thus my interest in countering the "lower lows" of NAD+ that probably occur with ageing.

 

(ps - I don't pay any attention to, nor do I give, any of the ratings.   The ratings system probably does more harm than good with respect to promoting scientific discussion.  imho)

 

Edit:  Another thing worth noting, imho, is that evolution is probably out to kill us older folks, not keep us alive (it needs space for new genes, etc., to compete with other species).  So there's a limit to the notion that if we just work with mother nature, then (as individuals) we'll be better off.  Rather, we'll need to break the rules in clever ways to beat its limitations.

Edited by warner, 03 November 2016 - 08:48 AM.

[stefan_001]

 

My reasoning for this one is quite different, a cell in trouble will be in trouble whether you sleep or not. The circadian rhythm is a system behavior like many other functions of the body and without doubt has a purpose. The question is probably more does NR supplementation mess up the rhythm and as result something else get negatively impacted. But for the individual cell health I dont see we need to maintain it.

 

(ps the rating of you post is not mine)

 

When I was younger, that's the way I thought about it too.  But I learned the hard way that older mammals have problems that younger mammals don't, and that nighttime in particular can be dangerous for them.  In my case, calorie restriction (and some occasional apnea) got me in trouble with glaucoma and a PED (pigment epithelial detachment), both of which are accelerated by low oxygen, low glucose (relative hypoglycemia), low blood pressure (i.e., ischemia, hypoxia) which were occurring at night while sleeping.  So the paper I cited a few posts back about NR being helpful in protecting retinal cells from such conditions was of special interest.  But I think in general nighttime can be dangerous in older mammals, in that the drop in nutrient levels and blood pressure, combined with generally lowered NAD+ (and melatonin, etc.) with ageing, can put at risk any subsystem (eyes, heart, brain, etc.) that happens to be particularly susceptible to such conditions.  Thus my interest in countering the "lower lows" of NAD+ that probably occur with ageing.

 

(ps - I don't pay any attention to, nor do I give, any of the ratings.   The ratings system probably does more harm than good with respect to promoting scientific discussion.  imho)

 

Edit:  Another thing worth noting, imho, is that evolution is probably out to kill us older folks, not keep us alive (it needs space for new genes, etc., to compete with other species).  So there's a limit to the notion that if we just work with mother nature, then (as individuals) we'll be better off.  Rather, we'll need to break the rules in clever ways to beat its limitations.

 

I think we agree. Maybe I didnt word well what I was trying to say. I meant I do not think we need to be too religious with the circadian rhythm when thinking of how and when to supplement NR. I see nothing usefull in the NAD+ dips so avoiding they go so deep is a good approach jmho

 

BTW not sure what you define as younger......lab rats in my "experiment" are between 46-78 years old

[mrkosh1]

I think my NAD+ system is screwed up after re-gaining 50 pounds of fat after falling off a diet that had seemed like a lifestyle change. I've went from being highly energetic to tired, weak, lethargic, and mentally foggy. The three things that seem to help are exercise (good for NAD+), eating high protein meals (don't know if this is good for NAD+), and R-Alpha Lipoic Acid when it doesn't kick in my hypoglycemia. If I can ever start taking some NR, I think it may help (screwy financial issues plaguing me).

 

Anyway, the NR stuff is exciting to me, especially if we can find a way to maximize it to reverse aging.

 

There is a rumor that I heard for a long time. I wonder if anyone refuted it. I read a few people state that NR could only boost NAD+ in mammals and humans for a short period of time (maybe weeks) before the body would adapt and the NAD+ level would drop. Some of the great health results in animal models seems to refute this, IMO.

 

I'll be eager to see a long term study of the effect of NR on NAD+ in humans.

[warner]

I think we agree. Maybe I didnt word well what I was trying to say. I meant I do not think we need to be too religious with the circadian rhythm when thinking of how and when to supplement NR. I see nothing usefull in the NAD+ dips so avoiding they go so deep is a good approach jmho   BTW not sure what you define as younger......lab rats in my "experiment" are between 46-78 years old

oops... sorry about that...  Another interesting thing about glaucoma is that you can measure the loss of neurons (the nerve fiber layer, NFL) in the retina directly with OCT, and that there is a baseline loss rate affecting all of us.  iow, we will all get glaucoma (go blind) if we live long enough (the baseline rate just gets you there well past 100 years).  Similar to all of us getting dementia if we live long enough.  Seems reasonable to think that NAD+ (and melatonin, etc.) declining with age has something to do with that, and that this may be another way in which NAD supplementation extends healthspan.

 

jmho, 50+ is "old" in the sense that it is beyond the age that most people lived for most of our species history, meaning there was not much selection pressure beyond that age, and what happens to us after that seems somewhat messy and arbitrary, with lots of subsystems falling apart or becoming uncoordinated in unexpected ways.  Also true that death of such old people was necessary for evolution to compete with other species, although in the past it could rely on predators, trauma, and disease to do that job, without having to directly plan our demise with what we now view as diseases of ageing.

Edited by warner, 03 November 2016 - 02:12 PM.

[Oakman]

.....

 

There is a rumor that I heard for a long time. I wonder if anyone refuted it. I read a few people state that NR could only boost NAD+ in mammals and humans for a short period of time (maybe weeks) before the body would adapt and the NAD+ level would drop. Some of the great health results in animal models seems to refute this, IMO.

 

I'll be eager to see a long term study of the effect of NR on NAD+ in humans.

 

I think it is pretty self-evident from the little human data we do have that (after NR supplementation over time) a decrease in serum levels of metabolites is a fact. Looking at the graphs over time, and with the exception of MeNam, each show some decrease re: initial peak levels by the endpoint. And this is with 1G dosing, so something is happening, as surely there is plenty ingested to (theoretically anyway) keep levels elevated at initial peak, or even ever increasing over time...but they are not).  Adaptation toward high levels of substances introduced in the body is well known, why should NR be any different?

 

This unfortunately points us to the real unknown in the longevity equation IMHO..."Why do NAD+ levels drop as we age?" IOW "What process ultimately control these levels?"  The body's depletion of NAD+ over time is a symptom of something we don't understand. Supplementation with NR is symptomatic treatment, but not a true solution to effectively moderating decreasing NAD+ levels through the body's own mechanisms.

 

Extended data to show what happens over weeks, months to NAD+ levels (and others) under supplementation would be very helpful. I think it may be coming as more studies complete.

 

 

http://www.nature.co...mms12948-f2.jpg

 

A healthy 52-year-old male ingested 1,000 mg NC Cl daily for 1 week. PBMCs were prepared from blood collected before the first dose (0 h), at seven time points after the first dose (0.6, 1, 1.4, 2.7, 4.1, 7.7 and 8.1 h), before second dose (23.8 h) and 24 h after the seventh dose (167.6 h). Concentrations (a, NMN; b, NAD+; c, NADP+; d, Nam; e, MeNam; f, Me2PY; g, Me4PY; h, NAAD; i, ADPR) are with respect to whole blood volumes. The data indicate that the NAD+metabolome—with the exception of Nam and NAAD—is elevated by the 4.1 time point post ingestion. Whereas PBMC Nam was never elevated by NR, NAAD was elevated by the next time point. NAD+ and NAAD remained elevated 24 h after the last dose.

Edited by Oakman, 03 November 2016 - 02:12 PM.

[Oakman]

 

 

My reasoning for this one is quite different, a cell in trouble will be in trouble whether you sleep or not. The circadian rhythm is a system behavior like many other functions of the body and without doubt has a purpose. The question is probably more does NR supplementation mess up the rhythm and as result something else get negatively impacted. But for the individual cell health I dont see we need to maintain it.

 

(ps the rating of you post is not mine)

 

When I was younger, that's the way I thought about it too.  But I learned the hard way that older mammals have problems that younger mammals don't, and that nighttime in particular can be dangerous for them.  In my case, calorie restriction (and some occasional apnea) got me in trouble with glaucoma and a PED (pigment epithelial detachment), both of which are accelerated by low oxygen, low glucose (relative hypoglycemia), low blood pressure (i.e., ischemia, hypoxia) which were occurring at night while sleeping.  So the paper I cited a few posts back about NR being helpful in protecting retinal cells from such conditions was of special interest.  But I think in general nighttime can be dangerous in older mammals, in that the drop in nutrient levels and blood pressure, combined with generally lowered NAD+ (and melatonin, etc.) with ageing, can put at risk any subsystem (eyes, heart, brain, etc.) that happens to be particularly susceptible to such conditions.  Thus my interest in countering the "lower lows" of NAD+ that probably occur with ageing.

 

(ps - I don't pay any attention to, nor do I give, any of the ratings.   The ratings system probably does more harm than good with respect to promoting scientific discussion.  imho)

 

Edit:  Another thing worth noting, imho, is that evolution is probably out to kill us older folks, not keep us alive (it needs space for new genes, etc., to compete with other species).  So there's a limit to the notion that if we just work with mother nature, then (as individuals) we'll be better off.  Rather, we'll need to break the rules in clever ways to beat its limitations.

 

I think we agree. Maybe I didnt word well what I was trying to say. I meant I do not think we need to be too religious with the circadian rhythm when thinking of how and when to supplement NR. I see nothing usefull in the NAD+ dips so avoiding they go so deep is a good approach jmho

 

BTW not sure what you define as younger......lab rats in my "experiment" are between 46-78 years old

 

 

First, what we need is a study that shows levels, showing a comparison of the absolute levels of NAD+ between young and old individuals, including the circadian dips, . That would help in deciding levels of supplementation to return to youthful levels. Maybe it is out there, but I haven't seen it.

 

Second, I think dismissing the body's circadian level changes is a bit too premature. The rhythms are there for some reason, even if we do not understand what that may be. NR supplementation swamps the natural production, and therefore whatever these circadian changes are meant to accomplish. Is the rhythm some way to repair, then clean the system repeatedly? Is it a vital step in energy production within the cell that we are swamping with supplementation?  There's a lot to learn yet about what we are attempting to do with NR to better NAD+.

 

Until that time, I see one missing piece in this puzzle as pinning down more about the "time to effect" of NR supplementation in the body.  Some say take without food, some say take with food, or it doesn't matter. If taken with food, assimilation is slowed by the food mass envolved, obviously. The studies to date seem to show ~8 hr to peak blood levels, but that does seem excessively long. Does food make it even longer, or slower acting? Why so long for NR anyway, when many substances taken orally are effective in far shorter periods (minutes)?

 

MY idea would be to try and maximize the natural peak levels, but that means timing dosing based on speed of absorption and blood levels. These seem far from exact at this point, mostly just a best guess it seems. More study results should prove enlightening.

Edited by Oakman, 03 November 2016 - 02:45 PM.