179 replies to this topic

[Turnbuckle]

Experiment #3: Inducing Mitophagy with NR

Following personal Experiment #1 and Experiment #2, this experiment is an attempt to determine if a lower dose of NR (2625 mg / day) might induce mitophagy.

 

2016-12-04  Day 1. Begin taking 7 capsules (875 mg) of NR three times per day for a total of 2625 mg / day. I have not taken C60-OO or any supplements or medications for a two weeks.

2016-12-07  Day 4. I have noticed any symptoms or side effects since beginning this treatment other than an occasional sensation of flushing a few minutes after ingesting NR.

2016-12-08  Day 5. There are no noticeable symptoms.  

2016-12-09  Day 6. Last day of this trial.  

2016-12-12  Day 9. Unlike Experiments #1 and #2, during this trial my normally regular, quiet lifestyle was interrupted due to unexpected business travel followed on days 6 and 7 by physically demanding, days-long home repair.  During days 6-9, I experienced an unusual level of muscle ache and stiffness.  I can’t determine the extent which this treatment or the home repair might have contributed to these muscle ache symptoms, or even if this this dose was sufficient to induce mitophagy at all.  A follow up experiment #4 at the earlier higher dose of NR (3750 mg/day) with my normal lifestyle will help clarify this situation.  If NR has been effective in inducing mitophagy, I would expect that there would be fewer symptoms of muscle ache in an experiment #4 even at the higher dose.

 

Possibly your mitochondrial population had not recovered sufficiently by the time you did all that repair work. Perhaps baking soda would would have replenished them.

[Izan]

So far in a group buy scenario I have:

 

Nate-2004    10 grams

Prophets        25 grams

Lost69             15 grams

Richard hnry   15 grams

count me in for 10 grams please.

[Nate-2004]

So far in a group buy scenario I have:

 

Nate-2004    10 grams

Prophets        25 grams

Lost69             15 grams

Richard hnry   15 grams

 

Where's this coming from and how much is needed?

Edited by Nate-2004, 17 December 2016 - 02:51 PM.

[Oakman]

I've read through this thread from the beginning and it's interesting as I have recently also done a "zombie mitochondrial" flush, not using the protocol given in this thread. My 3-day flush did seem to have an effect on me, most noticeably loss of energy initially, and a rather disconcerting major increase in floaters in one eye, followed by a significant boost of energy and strength shortly thereafter upon resuming normal supplementation. But that's another story.

 

However, I see there is now interest in Urolithin A supplementation, resulting in a group buy.  I had the same interest in Urolithins and so researched and found a different solution than supplementing the substance directly, that is, using precursors Robuvit¹ & Pterostilbene to upregulate its production in the gut (someone had mentioned Pterostilbene early in the thread I see).

 

¹ Robuvit  http://www.robuvit.com/faq/
                   http://www.robuvit.com/fileadmin/robuvit/robuvit_brochure_161_WEB.pdf

 

One of Robuvit's clinical trials, concerning those with "Chronic Fatigue Syndrome", indicated that Robuvit @200 mg/day basically got the job done in rejuvenating energy production in the body. Summarized here in this LEF article here > http://www.lifeextension.com/magazine/2014/12/new-option-for-chronic-fatigue-syndrome/page-01.

 

 

Another study, An Effect of Oak-Wood Extract (Robuvit®) on Energy State of Healthy Adults—A Pilot Study, showed similar results for healthy individuals, http://onlinelibrary.wiley.com/doi/10.1002/ptr.5368/pdf

 

-----

Again, according to the same LEF summary article, Robuvit increased plasma total phenols 100%, with ribosomal activity thru gene expression also noted. This increase in phenols was also what pomegranate extract/juice was trying to do, but here it is a result of direct Robuvit supplementation.

 

"Roburins In The Human Body 

 

 

 

"In the first study, researchers were interested in understanding how roburin molecules were distributed and absorbed, as well as their compatibility in the human body. Following five days of oral supplementation with roburin-rich oak wood extract—three capsules of a proprietary, patented extract called Robuvit®—the scientists found a 100% increase in plasma total phenols (a general measure of absorption of molecules in this class), as well as the presence of roburin breakdown products (metabolites) in urine of healthy volunteers.

 

Since roburins are found only in oak wood, the data demonstrated vigorous absorption and conversion of roburins into substances includingurolithins and ellagic acid, which are known to have potent biological activities.

This study also revealed that the oak wood roburins trigger a complex set of biological events in the body. Using a sophisticated technology that measures changes in gene expression, the researchers were able to show that blood serum from supplemented people in the study may beneficially alter the expression of several genes in human cells in culture.

Among the most consistent changes in gene expression induced by the serum from oak wood extract in supplemented patients had to do with the activities of ribosomes, the ultramicroscopic cellular organelles that are responsible for the “translation” of genes in DNA into specific proteins. Long regarded as simply tiny protein-manufacturing plants, ribosomes are now emerging as essential in the maintenance of normal cellular functions, and as key players in the science of “systemic aging” and disorders such as chronic fatigue syndrome."

 

------

Keep in mind ribosomes² are believe is being upregulated in this study, not mitophagy. Nevertheless, references in this thread show Urolithin does in fact, induce mitophagy.

 

²http://www.biology4k...cell_ribos.html > " Ribosomes are found in many places around a eukaryotic cell. You might find them floating in the cytosol. Those floating ribosomes make proteins that will be used inside of the cell. Other ribosomes are found on the endoplasmic reticulum. Endoplasmic reticulum with attached ribosomes is called rough ER. It looks bumpy under a microscope. The attached ribosomes make proteins that will be used inside the cell and proteins made for export out of the cell. There are also ribosomes attached to the nuclear envelope. Those ribosomes synthesize proteins that are released into the perinuclear space."

 

------

The main point of interest, is that Robuvit is readily available at $3.33/gram or less (Swanson), often 15% off @ $2.80/gram, and a 200-300mg dose is easily accomplished at a reasonable price. That compares more than favorably to the last best price for Urolithin A mentioned here @ $19.50/gram.

 

Perhaps this may offer an alternative for those who like what Urolithins potentially do, but not at the price of the single Urolithin A.

 

Anecdotally, I can't tell you if Robuvit plus Pterostilbene does what pure Urolithin A does (as I take too many things to be able to know), but since I started taking this combo of precursors, what I can mention is, some of my weights at the gym have almost magically gone up ~20+%, from 25lb to 30lb biceps curl, and 140lb to 180lbs seated high row, and 120lb to 150lbs Lat pulldown - in just a couple weeks’ time. After nearly a year of gym work, this increase in so short a time was astounding to me, whether autophagy and/or Urolithins or something else is helping. Notably, that increase does seem in line with the statistics given in the Robovit brochure .pdf (page 5 and also here³ showing significantly enhanced physical performance.

 

³ http://www.nutraingr...ete-performance

[Female Scientist]

Thanks Oakman. I'm wondering if you'd be willing to share your "zombie mitochondrial flush" protocol with us? Plus, more about whether you'd recommend it, and whether you follow it regularly? Mahalo.

I've read through this thread from the beginning and it's interesting as I have recently also done a "zombie mitochondrial" flush, not using the protocol given in this thread. My 3-day flush did seem to have an effect on me, most noticeably loss of energy initially, and a rather disconcerting major increase in floaters in one eye, followed by a significant boost of energy and strength shortly thereafter upon resuming normal supplementation. But that's another story.

 

However, I see there is now interest in Urolithin A supplementation, resulting in a group buy.  I had the same interest in Urolithins and so researched and found a different solution than supplementing the substance directly, that is, using precursors Robuvit¹ & Pterostilbene to upregulate its production in the gut (someone had mentioned Pterostilbene early in the thread I see).

 

¹ Robuvit  http://www.robuvit.com/faq/
                   http://www.robuvit.com/fileadmin/robuvit/robuvit_brochure_161_WEB.pdf

 

One of Robuvit's clinical trials, concerning those with "Chronic Fatigue Syndrome", indicated that Robuvit @200 mg/day basically got the job done in rejuvenating energy production in the body. Summarized here in this LEF article here > http://www.lifeextension.com/magazine/2014/12/new-option-for-chronic-fatigue-syndrome/page-01.

 

 

Another study, An Effect of Oak-Wood Extract (Robuvit®) on Energy State of Healthy Adults—A Pilot Study, showed similar results for healthy individuals, http://onlinelibrary.wiley.com/doi/10.1002/ptr.5368/pdf

 

-----

Again, according to the same LEF summary article, Robuvit increased plasma total phenols 100%, with ribosomal activity thru gene expression also noted. This increase in phenols was also what pomegranate extract/juice was trying to do, but here it is a result of direct Robuvit supplementation.

 

"Roburins In The Human Body 

 

 

 

"In the first study, researchers were interested in understanding how roburin molecules were distributed and absorbed, as well as their compatibility in the human body. Following five days of oral supplementation with roburin-rich oak wood extract—three capsules of a proprietary, patented extract called Robuvit®—the scientists found a 100% increase in plasma total phenols (a general measure of absorption of molecules in this class), as well as the presence of roburin breakdown products (metabolites) in urine of healthy volunteers.

 

Since roburins are found only in oak wood, the data demonstrated vigorous absorption and conversion of roburins into substances includingurolithins and ellagic acid, which are known to have potent biological activities.

This study also revealed that the oak wood roburins trigger a complex set of biological events in the body. Using a sophisticated technology that measures changes in gene expression, the researchers were able to show that blood serum from supplemented people in the study may beneficially alter the expression of several genes in human cells in culture.

Among the most consistent changes in gene expression induced by the serum from oak wood extract in supplemented patients had to do with the activities of ribosomes, the ultramicroscopic cellular organelles that are responsible for the “translation” of genes in DNA into specific proteins. Long regarded as simply tiny protein-manufacturing plants, ribosomes are now emerging as essential in the maintenance of normal cellular functions, and as key players in the science of “systemic aging” and disorders such as chronic fatigue syndrome."

 

------

Keep in mind ribosomes² are believe is being upregulated in this study, not mitophagy. Nevertheless, references in this thread show Urolithin does in fact, induce mitophagy.

 

²http://www.biology4k...cell_ribos.html > " Ribosomes are found in many places around a eukaryotic cell. You might find them floating in the cytosol. Those floating ribosomes make proteins that will be used inside of the cell. Other ribosomes are found on the endoplasmic reticulum. Endoplasmic reticulum with attached ribosomes is called rough ER. It looks bumpy under a microscope. The attached ribosomes make proteins that will be used inside the cell and proteins made for export out of the cell. There are also ribosomes attached to the nuclear envelope. Those ribosomes synthesize proteins that are released into the perinuclear space."

 

------

The main point of interest, is that Robuvit is readily available at $3.33/gram or less (Swanson), often 15% off @ $2.80/gram, and a 200-300mg dose is easily accomplished at a reasonable price. That compares more than favorably to the last best price for Urolithin A mentioned here @ $19.50/gram.

 

Perhaps this may offer an alternative for those who like what Urolithins potentially do, but not at the price of the single Urolithin A.

 

Anecdotally, I can't tell you if Robuvit plus Pterostilbene does what pure Urolithin A does (as I take too many things to be able to know), but since I started taking this combo of precursors, what I can mention is, some of my weights at the gym have almost magically gone up ~20+%, from 25lb to 30lb biceps curl, and 140lb to 180lbs seated high row, and 120lb to 150lbs Lat pulldown - in just a couple weeks’ time. After nearly a year of gym work, this increase in so short a time was astounding to me, whether autophagy and/or Urolithins or something else is helping. Notably, that increase does seem in line with the statistics given in the Robovit brochure .pdf (page 5 and also here³ showing significantly enhanced physical performance.

 

³ http://www.nutraingr...ete-performance

 

[Oakman]

 

Thanks Oakman. I'm wondering if you'd be willing to share your "zombie mitochondrial flush" protocol with us? Plus, more about whether you'd recommend it, and whether you follow it regularly? Mahalo.

 

 

 

Female Scientist, I'd certainly be willing to share the protocol. But, I've used it only once. Give me a couple weeks, I want to refine it a bit , and confirm results with a 2nd go.  

[Turnbuckle]

I am interested in this thread's topic of mitochondrial autophagy from the perspective of possible anti-cancer applications.

Many of the comments posted to this thread would seem to have anti-cancer potential. For example, the idea of changing

the NAD+/NADH ratio is noted in the cancer literature and on this thread it is specifically related to a signal to induce mitophagy.  

 

Might someone comment on how an anti-mitochondrial strategy might be combined with other anti-cancer strategies?

 

As an example, the article below found that ccRCC cells had a pronounced reduction in mitochondria and were more vulnerable to treatment with 3-BP. Might a combination of anti-mitochondrials and anti-glycolytics make for a sensible anti-cancer treatment? 

 

 

https://www.ncbi.nlm...les/PMC4669744/

 

Glycolytic inhibitors like bromopyruvate sound interesting. It apparently works because of the Warburg effect, where cells switch from oxidation to glycolysis, and thus they are starved by the glycolytic inhibitor. But since mitochondria are already shut down, I don't see the point of an anti-mitochondrial in general, especially as that might damage the entire organism. However, raising the NAD+/NADH ratio is actually good for the organism as it sets the quality control mechanism running and gets rid of defective mitochondria. Would it do that to cancer cells as well, and would that further starve those cells, at least temporarily? Seems possible. If so, then things like niacin and nicotinamide should reduce the incidence of cancer. And they do--

 

A Phase 3 Randomized Trial of Nicotinamide for Skin-Cancer Chemoprevention

 

CONCLUSIONS: Oral nicotinamide was safe and effective in reducing the rates of new nonmelanoma skin cancers and actinic keratoses in high-risk patients.

https://www.ncbi.nlm...pubmed/26488693

 

 

 

This was at a dose of 500 mg twice a day, which isn't much.  So using high-dose nicotinamide with a glycolytic inhibitor might be synergistic for attacking an existing cancer.

[mag1]

Thank you for your reply Turnbuckle!

 

I was thinking along the lines of eliminating the defective mitochondria that have the HK2 wrongly localized to VDAC.

The newly formed mitochondria through mitogenesis, should then be properly functioning(?), apoptosis(?), or at least

removing the HK2 ally might then be very helpful. Do an HK2 reset with the zombie mitochondria protocol!

 

Depriving cancer of its best friend

might rid humanity of its worst enemy.

 

 

​"A very common misconception... is that most cancerous tumors thrive exclusively off the sugar glucose via glycolysis for their energy source. In fact, as noted above, most frequently they derive significant amounts of their energy in the form of ATP from both the “Warburg effect”, as well as from mitochondrial oxidative phosphorylation. ...

 

 

It is now known that HK2 is highly expressed in most cancers whereas in most normal cells the predominant isoforms used are HK1, HK4, and to a lesser extent HK2 and HK3 ...

 

 

the mitochondrial binding of hexokinase (now known to be HK2) is bound to the outer membrane protein VDAC [40]. This localization of HK2 gives it preferred access to ATP produced during oxidative phosphorylation [41], and unlike other hexokinase isoforms, causes it to become insensitive to inhibition by its product, G-6-P [42,43].  ... Nevertheless, HK2 clearly stands out as the major contributor for the reasons already alluded to above. This and the fact that HK2 bound to mitochondrial VDAC [40] is also a major contributor to the immortalization of cancer cells [48,49] makes this isoenzyme one of cancer’s best friends. However, this also makes HK2 one of [cancer's] most vulnerable ... “Achilles heel” and therefore it is a highly promising drug target.

 

It is also worth mentioning that numerous mutations present in each tumor might come as a secondary side effect of metabolic changes related to the cancer disease. They arise after the initiation of uncontrolled growth due to damaged mitochondria [50]."

 

http://www.mdpi.com/...3049/21/12/1730

Edited by mag1, 20 December 2016 - 01:31 AM.

[Turnbuckle]

This discussion has turned to suppressing mitochondria in cancer cells (along with suppressing glucose metabolism), but cancer cells (the majority) already suppress mitochondria. This seems important to achieving immortality, just as it is in stem cells, which also suppress mitochondria. Culling non-functional mitochondria with NAD+ supplements seems to suppress cancer by a significant but not terribly interesting degree, but actually increasing mito number would return cancer cells to normal functioning, at least temporarily, where mito mediated processes could kill them--apoptotic cell death. A less-than-convincing thread discusses the possibility that bicarbonate might increase mito number, and if it does that in cancer cells, it would definitely have an impact. One doctor (Tullio Simoncini) has used bicarbonate for cancer treatment--though with another and certainly crackpot rationale. Other alkaline materials, such as cesium chloride, have been used for this purpose as well. 

Edited by Turnbuckle, 20 December 2016 - 11:48 AM.

[stefan_001]

This discussion has turned to suppressing mitochondria in cancer cells (along with suppressing glucose metabolism), but cancer cells (the majority) already suppress mitochondria. This seems important to achieving immortality, just as it is in stem cells, which also suppress mitochondria. Culling non-functional mitochondria with NAD+ supplements seems to suppress cancer by a significant but not terribly interesting degree, but actually increasing mito number would return cancer cells to normal functioning, at least temporarily, where mito mediated processes could kill them--apoptotic cell death. A less-than-convincing thread discusses the possibility that bicarbonate might increase mito number, and if it does that in cancer cells, it would definitely have an impact. One doctor (Tullio Simoncini) has used bicarbonate for cancer treatment--though with another and certainly crackpot rationale. Other alkaline materials, such as cesium chloride, have been used for this purpose as well. 

 

The theory that mitochondria are damaged in cancer is disproven. The aerobic glycosis also takes place when health cells proliferate.

[Turnbuckle]

 

 

 

The theory that mitochondria are damaged in cancer is disproven. The aerobic glycosis also takes place when health cells proliferate.

 

 

It's disproven? If so you should supply a link, as the following from last year suggests the opposite--

 

Restoration of mitochondria function as a target for cancer therapy

 

Defective oxidative phosphorylation has a crucial role in the attenuation of mitochondrial function, which confers therapy resistance in cancer. Various factors, including endogenous heat shock proteins (HSPs) and exogenous agents such as dichloroacetate, restore respiratory and other physiological functions of mitochondria in cancer cells. Functional mitochondria might ultimately lead to the restoration of apoptosis in cancer cells that are refractory to current anticancer agents. Here, we summarize the key reasons contributing to mitochondria dysfunction in cancer cells and whether and/or how restoration of mitochondrial function could be exploited for cancer therapeutics...

 

Recently, there has been renewed interest in targeting and restoring mitochondrial steady state as an attractive strategy for cancer control and management [20]. Metabolic inhibitors, including 2-deoxy-D-glucose (2DG), dichloroacetate (DCA), hexokinase inhibitors, and lactate dehydrogenase (LDH) inhibitors, have been used to specifically block aerobic glycolytic pathway and restore steady-state OXPHOS, and have been shown to be effective against various cancers in vitro and in vivo (Figure 1) [16]. In this regard, DCA has shown positive outcomes by inhibiting growth and proliferation of various cancers in vitro and in vivo by inducing cell cycle arrest and apoptosis.

 

https://www.ncbi.nlm...les/PMC4433775/

 

[stefan_001]

 

 

 

 

The theory that mitochondria are damaged in cancer is disproven. The aerobic glycosis also takes place when health cells proliferate.

 

 

It's disproven? If so you should supply a link, as the following from last year suggests the opposite--

 

Restoration of mitochondria function as a target for cancer therapy

 

Defective oxidative phosphorylation has a crucial role in the attenuation of mitochondrial function, which confers therapy resistance in cancer. Various factors, including endogenous heat shock proteins (HSPs) and exogenous agents such as dichloroacetate, restore respiratory and other physiological functions of mitochondria in cancer cells. Functional mitochondria might ultimately lead to the restoration of apoptosis in cancer cells that are refractory to current anticancer agents. Here, we summarize the key reasons contributing to mitochondria dysfunction in cancer cells and whether and/or how restoration of mitochondrial function could be exploited for cancer therapeutics...

 

Recently, there has been renewed interest in targeting and restoring mitochondrial steady state as an attractive strategy for cancer control and management [20]. Metabolic inhibitors, including 2-deoxy-D-glucose (2DG), dichloroacetate (DCA), hexokinase inhibitors, and lactate dehydrogenase (LDH) inhibitors, have been used to specifically block aerobic glycolytic pathway and restore steady-state OXPHOS, and have been shown to be effective against various cancers in vitro and in vivo (Figure 1) [16]. In this regard, DCA has shown positive outcomes by inhibiting growth and proliferation of various cancers in vitro and in vivo by inducing cell cycle arrest and apoptosis.

 

https://www.ncbi.nlm...les/PMC4433775/

 

 

 

here is one:

Conclusions

All tumor cell types show an enhanced glycolytic flux; however, not all have a diminished mitochondrial metabolic capacity. Therefore, the take-home message is that not all tumor cell types depend exclusively on glycolysis for ATP supply; some may equally or predominantly rely on oxidative phosphorylation. In consequence, the driving force for the enhanced glycolysis in tumor cells cannot be an energy deficiency induced only by a damaged oxidative phosphorylation.

 

http://onlinelibrary...07.05686.x/full

 

 

Edited by stefan_001, 24 December 2016 - 10:18 PM.

[Turnbuckle]

 

 

 

 

here is one:

Conclusions

All tumor cell types show an enhanced glycolytic flux; however, not all have a diminished mitochondrial metabolic capacity. Therefore, the take-home message is that not all tumor cell types depend exclusively on glycolysis for ATP supply; some may equally or predominantly rely on oxidative phosphorylation. In consequence, the driving force for the enhanced glycolysis in tumor cells cannot be an energy deficiency induced only by a damaged oxidative phosphorylation.

 

http://onlinelibrary...07.05686.x/full

 

 

 

Which is what I said, that "cancer cells (the majority) already suppress mitochondria." Thus nothing is disproven.

Edited by Turnbuckle, 24 December 2016 - 10:59 PM.

[stefan_001]

 

 

 

 

 

here is one:

Conclusions

All tumor cell types show an enhanced glycolytic flux; however, not all have a diminished mitochondrial metabolic capacity. Therefore, the take-home message is that not all tumor cell types depend exclusively on glycolysis for ATP supply; some may equally or predominantly rely on oxidative phosphorylation. In consequence, the driving force for the enhanced glycolysis in tumor cells cannot be an energy deficiency induced only by a damaged oxidative phosphorylation.

 

http://onlinelibrary...07.05686.x/full

 

 

 

Which is what I said, that "cancer cells (the majority) already suppress mitochondria." Thus nothing is disproven.

 

 

The way understand it is that the glycosis serves more the needs for proliferation building blocks than that it is needed for ATP generation. The article you posted was interesting, I read the the part of restoring mitochondrial function basicly that first we restore respiration so that when ROS is increased that activates apoptosis. But ROS increase already kills cancer cells. Perhaps I didn't extract the right parts of the article.

[stefan_001]

 

Experiment #2: Inducing Mitophagy with NR

The improvements to my skin from personal Experiment #1 have persisted. It is clear at this point that the benefits from that experiment are not transitory.  

 

This experiment repeats Experiment #1 to determine the extent further improvement is possible.

 

2016-10-19  Day 1. Begin taking 10 capsules (1250 mg) three times per day. I have not taken C60-OO or any supplements or medications  for a two weeks.

2016-10-20  Day 2. Like Experiment #1, my face feels very slightly flushed, as if I were taking a niacin.  There is no apparent difference in my skin.

2016-10-21  Day 3. Mid-day: My muscles feel more toned than usual but not stronger.  There is a slight feeling of flushing soon after I take NR.  Otherwise, I feel no symptoms.  PM: the skin on my arms appears smoother.

2016-10-22  Day 4. There is continued improvement to the skin on my arms, with fewer bumps and less discoloration. I feel normal, without muscle aches or other symptoms.

2016-10-23  Day 5. I feel colder and low energy, and otherwise without symptoms.  

2016-10-24  Day 6. Feeling a bit low energy and possibly have some weakness in my legs, but otherwise about normal.  There is no muscle ache like there was with Experiment #1. Remarkably, many of the smaller stucco keratoses on my hands and wrists have disappeared. The larger ones are flatter.  I noticed some recession of these keratoses in Experiment #1 but was not certain and didn't document it. I have had these keratoses for about 20 years; they resisted all attempts at treatment until now. Incidentally, my skin still has about the same degree of fine wrinkles and crepe-like appearance as before, from which I can infer that large doses of NR have little if any effect on collagen production.

2016-10-25  Day 7.  Yesterday was the last day of the treatment.  I’m pleased with the improvement to the appearance of my skin.  Today I feel good and more energetic.  IMHO, there is no benefit in following this treatment with PQQ.

2016-10-26  Day 8.  Following the pattern of Experiment #1, I feel normal.  There is substantial improvement to the appearance of skin on my hands, wrists and arms.  On close inspection, while the larger stucco keratoses on my hands are still present, they are flatter than before. 

2016-10-27  Day 9. I feel  normal.

 

The effects of NR are so wide reaching that I can't see that it is possible to assign these observations positively to mitophagy.  However, low energy is consistent with a reduction in mitochondrial mass.

 

 

So I bought some extra cans of NR to replicate this. With 46 years old I expect less impact, but old enough to get some improvements. Before going ahead may I please ask what of the improvements you noticed are lasting till today?
 

Edited by stefan_001, 02 January 2017 - 10:02 PM.

[Fafner55]

 

 

Experiment #2: Inducing Mitophagy with NR

The improvements to my skin from personal Experiment #1 have persisted. It is clear at this point that the benefits from that experiment are not transitory.  

 

This experiment repeats Experiment #1 to determine the extent further improvement is possible.

 

2016-10-19  Day 1. Begin taking 10 capsules (1250 mg) three times per day. I have not taken C60-OO or any supplements or medications  for a two weeks.

2016-10-20  Day 2. Like Experiment #1, my face feels very slightly flushed, as if I were taking a niacin.  There is no apparent difference in my skin.

2016-10-21  Day 3. Mid-day: My muscles feel more toned than usual but not stronger.  There is a slight feeling of flushing soon after I take NR.  Otherwise, I feel no symptoms.  PM: the skin on my arms appears smoother.

2016-10-22  Day 4. There is continued improvement to the skin on my arms, with fewer bumps and less discoloration. I feel normal, without muscle aches or other symptoms.

2016-10-23  Day 5. I feel colder and low energy, and otherwise without symptoms.  

2016-10-24  Day 6. Feeling a bit low energy and possibly have some weakness in my legs, but otherwise about normal.  There is no muscle ache like there was with Experiment #1. Remarkably, many of the smaller stucco keratoses on my hands and wrists have disappeared. The larger ones are flatter.  I noticed some recession of these keratoses in Experiment #1 but was not certain and didn't document it. I have had these keratoses for about 20 years; they resisted all attempts at treatment until now. Incidentally, my skin still has about the same degree of fine wrinkles and crepe-like appearance as before, from which I can infer that large doses of NR have little if any effect on collagen production.

2016-10-25  Day 7.  Yesterday was the last day of the treatment.  I’m pleased with the improvement to the appearance of my skin.  Today I feel good and more energetic.  IMHO, there is no benefit in following this treatment with PQQ.

2016-10-26  Day 8.  Following the pattern of Experiment #1, I feel normal.  There is substantial improvement to the appearance of skin on my hands, wrists and arms.  On close inspection, while the larger stucco keratoses on my hands are still present, they are flatter than before. 

2016-10-27  Day 9. I feel  normal.

 

The effects of NR are so wide reaching that I can't see that it is possible to assign these observations positively to mitophagy.  However, low energy is consistent with a reduction in mitochondrial mass.

 

 

So I bought some extra cans of NR to replicate this. With 46 years old I expect less impact, but old enough to get some improvements. Before going ahead may I please ask what of the improvements you noticed are lasting till today?
 

 

 

It is hard to objectively report on lasting benefits from an n=1 experiment.  While I can't rule out other factors, my best opinion is that I have at least as much energy as before these tests and my skin is smoother than it has been in years.  Collagen production was not affected and I have the same degree of crepy skin on the inside of my forearms.

 

Regarding possible toxicity, I had a standard comprehensive blood panel test a few days after experiment #2.  The results were not available when we discussed possible toxicity and ALT.  All measurements were within normal range.  My ALT was 13 U/L, well within the reference range of 9-46.

 

I still plan to do an experiment #4 at my earlier higher dose of NR (3750 mg/day) but have not been able to work it in my schedule.

[Valijon]

I'm if this has been asked before so here goes. What's the difference in energy levels with NR in comparison to niacinimide. I know there's a slight chemical variation. Theses a lot of hype around NR and I'm concerned that this is much of what NR is turning out to be.

I've watched supplement companies hype a product for decades at drastically high prices. Here at Longecity, we should be intelligent enough and we'll educated enough to not fall for hype. Im following the discussions surrounding NR but, I'm refraining from purchasing any NR until the price comes down. Its atrociously high at the moment.

[stefan_001]

 

 

 

Experiment #2: Inducing Mitophagy with NR

 

 

 

So I bought some extra cans of NR to replicate this. With 46 years old I expect less impact, but old enough to get some improvements. Before going ahead may I please ask what of the improvements you noticed are lasting till today?
 

 

 

It is hard to objectively report on lasting benefits from an n=1 experiment.  While I can't rule out other factors, my best opinion is that I have at least as much energy as before these tests and my skin is smoother than it has been in years.  Collagen production was not affected and I have the same degree of crepy skin on the inside of my forearms.

 

Regarding possible toxicity, I had a standard comprehensive blood panel test a few days after experiment #2.  The results were not available when we discussed possible toxicity and ALT.  All measurements were within normal range.  My ALT was 13 U/L, well within the reference range of 9-46.

 

I still plan to do an experiment #4 at my earlier higher dose of NR (3750 mg/day) but have not been able to work it in my schedule.

 

 

Thanks for that update appreciated. Good to hear the ALT looked fine. Ok I will plan a test and write the perceptions in this thread.

[stefan_001]

I'm pretty skeptical about NR inducing mitophagy or biogenesis and even if the NAD ratio is relevant, NR doses beyond 300mg have a diminishing returns on NAD+. Taking that much NR is likely a waste of money if this is what you're trying to achieve. Plus you get a 50% boost in NAD+ from 300mg alone that lasts most of the day, peaking at specific times. 

 

PQQ has some preliminary evidence for humans. There is so little research on PQQ, no controlled or double blind tests to be sure because preliminary evidence showed little promise for a lot of specific conditions being treated. There are several diseases theorized to be related to mitochondrial dysfunction including the one I have, Essential Tremor, but this theory is relatively recent. 

 

I'm considering taking some but worried it's just throwing my money down the drain yet again.

 

Hi Nate, true the stats show a diminishing return for the periods measured. It may also be that it is that long tail that has the extra impact. The summary on this webpage show interesting effect achieved at high dosing:

 

http://www.timelessl...osing-overview/

 

At least it will not hurt (and now we know in addition from Fafner55 that liver is not a worry). So an overflooding experiment is certainly interesting to try.

 

[Turnbuckle]

...I've watched supplement companies hype a product for decades at drastically high prices. Here at Longecity, we should be intelligent enough and we'll educated enough to not fall for hype. Im following the discussions surrounding NR but, I'm refraining from purchasing any NR until the price comes down. Its atrociously high at the moment.

 

 

I agree, and assuming that NR is better than NAM (not proven), I think one can probably add ribose to NAM and let your cells do the chemistry. It would also be interesting to add quercetin and/or apigenin, which are known to conserve NAD+. All of these things are really cheap.

 

 

Flavonoid Apigenin Is an Inhibitor of the NAD+ase CD38

 

we characterize two CD38 inhibitors: quercetin and apigenin. We show that pharmacological inhibition of CD38 results in higher intracellular NAD+ levels

 

Edited by Turnbuckle, 03 January 2017 - 05:51 PM.

[stefan_001]

I have added Quercetin for some time to the NR I use for that reason. It is hard to judge whether it helps or not as I also supplement other polyphenols and also because you loose reference fast. My initial  impressions however were positive.

 

For the high dose NR regime I was thinking whether it would make sense to take Q at the same time. Another one would be fasting at the same time. Did anybody try fasting + mito regime?
 

[Nate-2004]

Since senescent cells (SASP) are what may be at the root of NAD+ depletion with age, drugs that target senescent cells should be the primary goal. Before you buy more NR or quercetin or dasatanib, please put some money into researching this drug. The crowdfund campaign for this research is $28,000 short with 3 days to go. Just 1300 more people donating $25 would be enough to help out.  Please share on your FB and share with everyone you know.

 

Link to the campaign.

 

Link to the official thread.

Edited by Nate-2004, 17 January 2017 - 06:18 PM.

[stefan_001]

Since senescent cells (SASP) are what may be at the root of NAD+ depletion with age, drugs that target senescent cells should be the primary goal. Before you buy more NR or quercetin or dasatanib, please put some money into researching this drug. The crowdfund campaign for this research is $28,000 short with 3 days to go. Just 1300 more people donating $25 would be enough to help out.  Please share on your FB and share with everyone you know.

 

Link to the campaign.

 

Link to the official thread.

 

Sounds rather unrealistic.

 

On a side note I have not yet done the NR experiment. Too busy working weeks.
 

[Nate-2004]

 

 

 

Sounds rather unrealistic.

 

On a side note I have not yet done the NR experiment. Too busy working weeks.
 

 

 

Meeting the goal or finding a drug that clears senescent cells? Meeting the goal in the last 3 days is not unrealistic I don't think. Many campaigns have seen big jumps at the last minute.

[stefan_001]

 

 

 

 

Sounds rather unrealistic.

 

On a side note I have not yet done the NR experiment. Too busy working weeks.
 

 

 

Meeting the goal or finding a drug that clears senescent cells? Meeting the goal in the last 3 days is not unrealistic I don't think. Many campaigns have seen big jumps at the last minute.

 

 

With 40k you don't get done too much, Probably all he will do is pay himself a salary and do some research. A far cry from drug development.

[sthira]

With 40k you don't get done too much, Probably all he will do is pay himself a salary and do some research. A far cry from drug development.

I wonder the same thing, actually, and think this is a fair question. Why don't you ask the researcher directly involved in this project, and see how he responds. Here: http://www.longecity...e-2#entry802647

[Nate-2004]

 

 

With 40k you don't get done too much, Probably all he will do is pay himself a salary and do some research. A far cry from drug development.

 

 

 

 

To quote from the project web page. $40,000: Initial Goal – Candidate promoter design

Our first work project is to design candidate synthetic promoters which would be able to accurately detect senescent cells. To do that, we will analyse transcription profiles of a number of senescent cells. To cover all different senescence states, we will analyse senescent cells from different tissues and at different stage of senescence. We will use this data and proprietary bioinformatics algorithms to design genetic circuits which are specific to these cells.

 

 

And

 

 

 

I believe the first stage of the project is to pay for lab-equipment time to develop markers they can target.

Edited by Nate-2004, 17 January 2017 - 06:59 PM.

[Fafner55]

This review article on the mitochondrial-lysosomal axis might be useful.

     Mitochondrial Turnover and Aging of Long-Lived Postmitotic Cells: The Mitochondrial–Lysosomal Axis Theory of Aging (2010)  

 

It should be possible to enhance mitophagy by supplementing with curcumin phytosome, green tea extract phytosome or sulforaphane.

1. Neuroprotective and anti-ageing effects of curcumin in aged rat brain regions (2006)   
2. Epigallocatechin gallate inhibits beta amyloid oligomerization in Caenorhabditis elegans and affects the daf-2/insulin-like signaling pathway (2010) 
3. Sulforaphane enhances progerin clearance in Hutchinson-Gilford progeria fibroblasts (2015) 

Besides phytosome formulations, 10 to 20 mg of the black pepper extract piperine is proven to improve the bio-availability of curcumin and green tea extract.

1. Piperine Enhances the Bioavailability of the Tea Polyphenol (−)-Epigallocatechin-3-gallate in Mice (2004)
2. Influence of piperine on the pharmacokinetics of curcumin in animals and human volunteers (1998)

 

 

[Fafner55]

Here are articles indicating that inducing mitophagy is more than just depolarizing mitochondria and activating lysosomes, as I tried to point out in my last post.  It is well  known that the PINK1/Parkin pathway is also involved and is activated by sulforaphane.

1. PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy (2014)
2. PINK1- and Parkin-mediated mitophagy at a glance (2012)

[Fafner55]

Experiment #4: Inducing Mitophagy with NR While Activating Autophagy

This experiment attempts to improve mitophagy by activating autophagy and reducing lipofuscin while depolarizing mitochondria. 

 

For 6 days on an empty stomach take

1. Nicotinamide riboside (NR), 1250 mg doses 3 times per day 
2. Centrophenoxine, 250 mg per day
3. Green tea extract phytosome, 250 mg, 2x per day 
4. Curcumin phytosome 500 mg, 2x per day
5. Sulforaphane glucosinolate, 2 capsules = 60 mg in 770 mg broccoli extract, 2x per day
6. Piperine, 10-15 mg, 2x/day

 

2017-02-02  Day 0. Took centrophenoxine, 250 mg.

2017-02-03  Day 1. Began taking the above described treatment.  Not much in the way of side effects.  Maybe more energy.

2017-02-04  Day 2. No symptoms or noticeable effect except the piperine slightly irritates my stomach.

2017-02-05  Day 3. No symptoms or noticeable effect except for fading of age spots.  When I previously self-tested centrophenoxine a couple of years ago, I noticed steady day-by-day fading of age spots on my hands. Long term use of centrophenoxine lead to feelings of brain fog from elevated acetylcholine levels.

2017-02-06  Day 4. No symptoms or noticeable effect except for continued fading of age spots.

        Day 4. PM.  I feel unusually tired.

2017-02-07  Day 5.  No symptoms or noticeable effect except for fading of age spots. I have good energy.

2017-02-08  Day 6.  This is the last day of the treatment.  There are no symptoms or noticeable effects.  I have good energy. This trail is different than Experiment #1 in that on day 6 I felt noticeably weak then but not now. I speculate that the absence of symptoms supports the idea that NR induces mitophagy preferentially in dysfunctional mitochondria.

2017-02-09  Day 7.  Work early feeling energetic. 

2017-02-10  Day 8.  I am alert and energetic.

2017-02-11  Day 9.  No changes to how I look and feel.

 

This trial is different than experiments #1 and #2.  In those experiments on day 6 I felt noticeably weak whereas in this one I felt nothing. The lack of effect of this experiment can be interpreted in at least two ways.

1. NR induces mitophagy preferentially in dysfunctional mitochondria which are sufficiently cleared at this point. My Experiment #3 supports this view since I noticed few if any symptoms then.
2. My well intentioned attempt at simulating autophagy possibly had the consequence of inhibiting mitophagy.  There is precedence for this effect in fasting. "Time-dependent differential effects of fasting on cardiac autophagy and mitophagy" (2016)  Results A 24-hour fasting increased mitophagy flux as demonstrated by a 44.2% increase in the number of mitophagy events in heart sections. The average size of mitophagy events within studied myocytes increased by 13.4%. Western blot analysis showed an increase in LC3-II protein levels in both total cardiac tissue lysates and the mitochondrial fractions, suggesting that autophagy and mitophagy were enhanced in parallel. However, after 48 hours of starvation, mitophagy events decreased by 50.1% compared to events at 24 hours of fasting and decreased by 28% compared to the control fed animals. The average area of each mitophagy event after 48 hours of fasting decreased by 39.9% compared to that at 24 hours and decreased by 31.9% compared to control. Interestingly, Western Blot analysis showed that LC3-II protein levels were increased in the total cardiac tissue lysates but reduced in the mitochondrial fractions, suggesting that the 48-hour fasting enhanced general autophagy but inhibited mitophagy.