459 replies to this topic

[niner]

Metabolism

After the liver, quercetin exists in the blood solely as quercetin glucuronides. Regardless of initial source, all forms of quercetin undergo hydrolysis and get glucuronidated in the liver before being released into systemic circulation.

 

Optimal Q concentrations for inducing senescent preadipocyte and HUVEC cell death
were 20 and 10 μM, respectively.

 

Herein lies the problem.  All those pharmacokinetic measurements must be "total quercetin", i.e. all the various glucuronides, sulfates, etc, along with whatever femtoscopic amount of the aglycone that escaped metabolism.  The in vitro results that found optimal concentrations of 20 and 10 uM, on the other hand, are the aglycone.  Thus the big questions--  Do any of the metabolites have senolytic activity?  Is quercetin metabolism in mice different than in humans?

[Logic]

 

Metabolism

After the liver, quercetin exists in the blood solely as quercetin glucuronides. Regardless of initial source, all forms of quercetin undergo hydrolysis and get glucuronidated in the liver before being released into systemic circulation.

 

Optimal Q concentrations for inducing senescent preadipocyte and HUVEC cell death
were 20 and 10 μM, respectively.

 

Herein lies the problem.  All those pharmacokinetic measurements must be "total quercetin", i.e. all the various glucuronides, sulfates, etc, along with whatever femtoscopic amount of the aglycone that escaped metabolism.  The in vitro results that found optimal concentrations of 20 and 10 uM, on the other hand, are the aglycone.  Thus the big questions--  Do any of the metabolites have senolytic activity?  Is quercetin metabolism in mice different than in humans?

 

 

That previous post was a balls-up with the forum software tossing my last piece of text Niner.

I finished my thoughts in a following post, but I suppose all the effort was for naught if its free aglycone that is the active ingredient?

Edited by Logic, 10 April 2015 - 01:57 PM.

[Fafner55]

Luteolin is another flavonoid, similar in structure to quercetin, that is known to induce apoptosis by activating the caspase cascade at similar plasma levels as quercetin.  “Pro-apoptotic effects of the flavonoid luteolin in rat H4IIE cells.” (2005)  ...we analysed DNA strand break formation by luteolin and found a distinct increase of DNA strand breaks after incubation for 3h with 100 microM luteolin, a concentration which induces oligonucleosomal DNA cleavage at 24h. In conclusion, the sequence of events is compatible with the assumption that luteolin triggers the mitochondrial pathway of apoptosis, probably by inducing DNA damage.  

 

Another study of luteolin’s caspase activation to induce apoptosis indicates doses feed to rats.  “Caspase activation and extracellular signal-regulated kinase/Akt inhibition were involved in luteolin-induced apoptosis in Lewis lung carcinoma cells“ (2007) states … In addition,  luteolin significantly inhibited the growth of LLC cells implanted on the flank of mice to 40% and 60% of untreated control group values at 2 mg/kg and 10 mg/kg, respectively.  These doses are 4% to 20% of the quercetin doses used to clear senescent cells in mice.

 

Large doses of luteolin (100 mg / 10 kg body weight) were given to children daily for 26 weeks without major adverse effects.  “An open-label pilot study of a formulation containing the anti-inflammatory flavonoid luteolin and its effects on behavior in children with autism spectrum disorders.” (2013)

Another study supports of luteolin for inducing apoptosis at the same concentration as quercetin.  (Table 1 of http://scholarworks.sjsu.edu/cgi/viewcontent.cgi?article=7788&context=etd_theses).  This dose is 20% of the quercetin doses used to clear senescent cells in mice.  Luteolin is said to have high bioavailability.

Taking an 8000 mg dose of quercetin dihydrate per day for two or three days appears to be a good starting point since there is large individual variability and few side effects.  Experimentation is needed to determine an effective dose.  Unfortunately, I do not currently have metabolic markers or observable measures of the effects of quercetin.  In addition to quercetin dihydrate, a 1600 mg dose of luteolin (20% of quercetin) can also be taken with reasonable safety.

 

 

My Experience with Clearing Senescent Cells

 

2015-03-13 to 15  I took 3200 mg of quercetin twice per day for three days last weekend along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg). Within days after my first treatment, the skin on my arms and body is noticeably smoother and clearer.  Now, three days later, my skin is clearer.  Age spots (raised brown areas) on my arms that I closely watch have noticeably faded, possibly due to the effects of 22 mg of C60-OO I took on 2015-03-15.  I will not take C60 again while this experiment is ongoing.

2015-03-20 Took pictures of my right hand and forearm for reference.

2015-03-22 I took 6400 mg quercetin all at once, along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg).

2015-03-24  The cells on the backs of my hand appear to be more uniform and smooth.  Age spots on my arms appear slightly smaller but are starting to darken.  It may have been the influence of C60-OO taken on 2015-03-16 that caused them to fade initially.

2015-03-27  I took 8000 mg of quercetin and 1600 mg of luteolin in a single dose.  An hour earlier I took  pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (1000 mg).   

At 3 hours, experienced slight muscle aches and a slight headache.

At 4 hours, I feel normal.

At 6 hours, an area of skin about 0.3 x 0.5 cm on my left hand is red.  Otherwise I look and feel normal.

2015-03-28  The redness on a patch of skin on my left hand has substantially receded.

2015-03-29  The redness on a patch of skin on my left hand is gone.  That area of skin appears different than normal skin; it appears to be a keratosis.

2015-03-29  Took pictures of my right hand and forearm and could not see a change in aged spots compared to 2015-03-20.  

2015-04-01  Icariin causes cells to resist apoptosis and will be eliminated from my senescent cell clearance treatment.  And, because of the side effects associated with the last treatment I am reducing luteolin to 800 mg.  

I took 8000 mg of quercetin, 800 mg of luteolin,  pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (625 mg) in a single dose.  I experienced no side effects.

2015-04-02  I took pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (625 mg) in the morning, followed by 8000 mg of quercetin and 800 mg of luteolin in the late afternoon.  I experienced no side effects.

2015-04-03  I could not be certain of the effect of NR on this experiment so I stopped taking it.  Today I took only 8000 mg of quercetin and 800 mg of luteolin.  I experienced no side effects.

2015-04-04  Today I took another dose of 8000 mg of quercetin and 800 mg of luteolin along with pterostilbene (100 mg) and resveratrol (500 mg).  I experienced no side effects.

2015-04-05  Took another dose of 8000 mg of quercetin and 800 mg of luteolin along with pterostilbene (100 mg) and resveratrol (500 mg).  Experienced lingering muscle fatigue and soreness in legs after a 2.5 mile walk.

At this point, if quercetin is effective, it should have done it’s work.  I  plan to do nothing further other than wait a few days to see if the raised brown areas on my arms recede.

2015-04-09  Took pictures of my right hand and forearm and could not see a change in aged spots or skin texture from the earlier pictures.

2015-04-10  My 59 year old skin looks the same as I did before beginning this treatment.  If senescent cells have been cleared from my body to any degree, I cannot observe an effect.

[Brett Black]

I haven't been following this area much, and my post is certainly not a recommendation for some of the experimentation suggested in this thread, but...

 

I happened to be recently watching a SENS video of Judith Campisi at Rejuvenation Biotechnology 2014, and reading through a bit of this thread I recalled the part at 5:27 - "One problem in the field is that there is actually nothing that I know of that is truly a sensescent specific marker. So we tend to look at a panapoly of markers in order to identify senescent cells."

 

Also, Judith talked about something that seemed troubling to me in regards to potential side effects of senolytics. Senescent cells may play an important and beneficial role in healing and suppressing senescent cells can apparently impair healing in mice - see 24:00 onwards:

 

 

 

Edited by Brett Black, 10 April 2015 - 03:15 PM.

[corb]

Also, Judith talked about something that seemed troubling to me in regards to potential side effects of senolytics. Senescent cells may play an important and beneficial role in healing and suppressing senescent cells can apparently impair healing in mice - see 24:00 onwards:

 

That concern has already been voiced and even though it is a valid point, it's also not really a possibility. None of the two drugs we know to remove senescent cells right now are effective enough to completely remove all senescent cells from your body. Not even in the tissues they are most effective in.
We know the old have a significantly higher amount of senescent cells than the young, so even if you remove a large amount, which you probably won't be able to do anyway with D+Q, you should still have enough for wound healing.

What I'm worried about is the people taking superdoses of quercetin, there's a possibility the will inadvertently find the amount where it stops being safe.
 

[Logic]

Quercetin activates Sirt1 in vivo

geddarkstorm:

"Yes indeed. That paper reported a plasma half life of 6-12 hours (that's some serious variability, heh). I've seen studies in humans that shows a plasma half life of 16 hours. It seems to reach a steady state, however, and taking some every day does not cause it to accrue beyond some point, depending on the repeated dosage, as we've seen in human studies."

http://www.longecity...-sirt1-in-vivo/

 

http://www.longecity...-sirt1-in-vivo/

 

Resveratrol and Quercetin combination

http://www.longecity...bination/page-2

[nowayout]

What I'm worried about is the people taking superdoses of quercetin, there's a possibility the will inadvertently find the amount where it stops being safe.

 

Quercetin affects a number of liver enzymes even at normally recommended doses, so anybody who is on any medications should be very careful.

 

Also, what concerns me about the shock dose idea is:  What are the likely side effects of killing off large numbers of senescent cells over the course of a day or two?  Wouldn't one expect at least a lot of inflammation and accompanying pain as a result, if not worse things?   
 

[niner]

Also, what concerns me about the shock dose idea is:  What are the likely side effects of killing off large numbers of senescent cells over the course of a day or two?  Wouldn't one expect at least a lot of inflammation and accompanying pain as a result, if not worse things?  

 

Probably not, in that the senolytics we have at the moment are not that good, most of us don't really have all that many senescent cells, and the cells are undergoing apoptosis rather than necrosis, so the ones that we do manage to kill will go out cleanly.  It's possible that we'd have a problem with extremely old people using potent senolytic agents.  That would be a situation where I would consider some sort of throttling approach, like brief exposures and/or low doses, with a recovery period in between doses.  I imagine that all of this will get worked out over time.  I just hope it doesn't take three decades.

[zorba990]

Wondering if rutin will work just as well. Bulk rutin is way cheaper than Quercetin (like 5x)
It does seem bioavailability of rutin may be less optimal, but still shows up as quercetin in the blood.
http://www.ncbi.nlm....pubmed/11151743

[aribadabar]

 

 

My Experience with Clearing Senescent Cells

 

2015-03-13 to 15  I took 3200 mg of quercetin twice per day for three days last weekend along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg). Within days after my first treatment, the skin on my arms and body is noticeably smoother and clearer.  Now, three days later, my skin is clearer.  Age spots (raised brown areas) on my arms that I closely watch have noticeably faded, possibly due to the effects of 22 mg of C60-OO I took on 2015-03-15.  I will not take C60 again while this experiment is ongoing.

....

2015-03-24  The cells on the backs of my hand appear to be more uniform and smooth.  Age spots on my arms appear slightly smaller but are starting to darken.  It may have been the influence of C60-OO taken on 2015-03-16 that caused them to fade initially.

........

 

2015-04-10  My 59 year old skin looks the same as I did before beginning this treatment.  If senescent cells have been cleared from my body to any degree, I cannot observe an effect.

 

I am left wit the impression that the test combo did nothing on the age spots and it was C60-oo's effect that faded over time due to the suspension of C60 intake.

 

Are you planning to continue the quercetin experiment or you have already decided to conclude it due to lack of apparent effects?

[DeadMeat]

Metabolism

After the liver, quercetin exists in the blood solely as quercetin glucuronides. Regardless of initial source, all forms of quercetin undergo hydrolysis and get glucuronidated in the liver before being released into systemic circulation.

 

Optimal Q concentrations for inducing senescent preadipocyte and HUVEC cell death
were 20 and 10 μM, respectively.

 

Herein lies the problem.  All those pharmacokinetic measurements must be "total quercetin", i.e. all the various glucuronides, sulfates, etc, along with whatever femtoscopic amount of the aglycone that escaped metabolism.  The in vitro results that found optimal concentrations of 20 and 10 uM, on the other hand, are the aglycone.  Thus the big questions--  Do any of the metabolites have senolytic activity?  Is quercetin metabolism in mice different than in humans?

 

Quercetin glucuronide can apparently get converted back locally into free quercetin aglycone, at inflammation/mitochondrial dysfunction sites, with help from macrophages. I wonder if that is relevant for senescent cells as well?

 

http://www.ncbi.nlm....les/PMC4042145/

J Clin Biochem Nutr. 2014 May;54(3):145-50. doi: 10.3164/jcbn.14-9. Epub 2014 Apr 12.
ß-Glucuronidase activity and mitochondrial dysfunction: the sites where flavonoid glucuronides act as anti-inflammatory agents.
Kawai Y.

Epidemiological and experimental studies suggest that the consumption of flavonoid-rich diets decreases the risk of various chronic diseases such as cardiovascular diseases. Although studies on the bioavailability of flavonoids have been well-characterized, the tissue and cellular localizations underlying their biological mechanisms are largely unknown. The development and application of novel monoclonal antibodies revealed that macrophages could be the major target of dietary flavonoids in vivo. Using macrophage-like cell lines in vitro, we examined the molecular basis of the interaction between the macrophages and flavonoids, especially the glucuronide metabolites. We have found that extracellular ß-glucuronidase secreted from macrophages is essential for the bioactivation of the glucuronide conjugates into the aglycone, and that the enzymatic activity, which requires an acidic pH, is promoted by the increased secretion of lactate in response to the mitochondrial dysfunction. This review describes our recent findings indicating the molecular mechanisms responsible for the anti-inflammatory activity of dietary flavonoids within the inflammation sites. We propose that the extracellular activity of ß-glucuronidase associated with the status of the mitochondrial function in the target cells might be important biomarkers for the specific sites where the glucuronides of dietary flavonoids can act as anti-atherosclerotic and anti-inflammatory agents in vivo.

I haven't fully read it yet, but the article above seems to be a review of this article:
http://www.ncbi.nlm....les/PMC3834324/

[Logic]

Quercetin glucuronide can apparently get converted back locally into free quercetin aglycone, at inflammation/mitochondrial dysfunction sites, with help from macrophages. I wonder if that is relevant for senescent cells as well?

I had a long post about why upregulate autophagy while doing senolytics was a good idea, but its gone now.
Probably about 6 hours worth of research, links, and typing; just gone...

Isn't it amazing how Google Docs and Gmail are able to save every keystroke..?

IP.Board, this forum software, does have an autosave feature:
http://community.inv...auto-save-post/
But until its enabled my reply has to be:

Yes they are related and it does seem to be a good idea to upregulate autophagy while doing Senolytics.

It seems to be tying into the NAD+ World too:

"...quercetin increases gene expression of the NQO1 gene via an Nrf2 transcription factor mediated pathway(ref)(ref)(ref). Specfically, quercetin enhances the binding of Nrf2 to the NRF-ARE binding site on the NQO1 gene promoter. Quercetin also increases Nrf2-mediated transcriptional activity by up regulation gut e expression of Nrf2 mRNA and Nrf2 protein. Quercetin also reduces the level of Keap1 protein, the binding partner of Nrf2, which prevents Nrf2 nuclear translocation. Quercetin reduces Keap-1in a post-translational mechanism, thereby reducing Nrf2 ubiquitination and proteasomal degradation..."
http://www.longecity...ted-aging-part/

[niner]

Quercetin glucuronide can apparently get converted back locally into free quercetin aglycone, at inflammation/mitochondrial dysfunction sites, with help from macrophages. I wonder if that is relevant for senescent cells as well?

 

It might be, because senescent cells have a tendency toward mitochondrial dysfunction and inflammation (due to the SASP).  Thanks for posting that paper.

[Florian Xavier]

chances that it work on humans ?

[Fafner55]

 

 

 

My Experience with Clearing Senescent Cells

 

2015-03-13 to 15  I took 3200 mg of quercetin twice per day for three days last weekend along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg). Within days after my first treatment, the skin on my arms and body is noticeably smoother and clearer.  Now, three days later, my skin is clearer.  Age spots (raised brown areas) on my arms that I closely watch have noticeably faded, possibly due to the effects of 22 mg of C60-OO I took on 2015-03-15.  I will not take C60 again while this experiment is ongoing.

....

2015-03-24  The cells on the backs of my hand appear to be more uniform and smooth.  Age spots on my arms appear slightly smaller but are starting to darken.  It may have been the influence of C60-OO taken on 2015-03-16 that caused them to fade initially.

........

 

2015-04-10  My 59 year old skin looks the same as I did before beginning this treatment.  If senescent cells have been cleared from my body to any degree, I cannot observe an effect.

 

I am left wit the impression that the test combo did nothing on the age spots and it was C60-oo's effect that faded over time due to the suspension of C60 intake.

 

Are you planning to continue the quercetin experiment or you have already decided to conclude it due to lack of apparent effects?

 

 

I took enough quercetin for a long enough period of time that it should have had an effect if it could have.  At my age, some of the various skin problems I have must be traceable to senescent cells, so if I am not seeing any sign of change I am skeptical that there is in fact a change.  Without apparent benefit, I have decided not to continue this experiment.  I will try quercetin with dasatinib if I can acquire some and after doing more research on why my experiment apparently failed.  For what I know, this experiment should have shown a positive result, even if the result was minor.  I am at a loss of why it didn't.

[Fafner55]

Wondering if rutin will work just as well. Bulk rutin is way cheaper than Quercetin (like 5x)
It does seem bioavailability of rutin may be less optimal, but still shows up as quercetin in the blood.
http://www.ncbi.nlm....pubmed/11151743

 

The bioavailability of rutin appears to be very poor.

Respective bioavailability of quercetin aglycone and its glycosides in a rat model.

A large number of flavonoids, mostly O-glycosides, are found in foods of plant origin. The bound sugar moiety is known to influence their bioavailability. We examined here the effect of the nature of the sugar on the absorption of the glycosides. Four groups of rats (n = 6) received a meal containing 20 mg of quercetin equivalents supplied as aglycone, quercetin 3-glucoside, quercetin 3-rhamnoside or rutin. Plasma were hydrolysed by a beta-glucuronidase/sulfatase and analyzed by HPLC coupled to UV detection at 370 nm. Four hours after the beginning of the meal, the quercetin metabolites present in plasma were identical in all groups but their total concentrations were quite different. With pure quercetin the circulating levels were 1.7 +/- 1.8 microM, but this level was three fold higher when quercetin was supplied as quercetin 3-glucoside (33.2 +/- 3.5 microM). By contrast, the plasma concentrations of quercetin metabolites was quite low with the rutin meal (about 3 microM) and undetectable after the quercetin 3-rhamnoside meal. These data suggest that the 3-O-glucosylation improves the absorption of quercetin in the small intestine, whereas the binding of a rhamnose or of a glucose-rhamnose moiety to the aglycone markedly depressed its absorption. Additionnal experiments have shown that the higher plasma levels measured after the meal containing quercetin 3-glucoside compared to quercetin were maintained throughout a 24 hour period following the meal. In conclusion, the nature of the glycosylation markedly influences the efficiency of quercetin absorption in rats. Quercetin 3-glucose can be absorbed in the small intestine and is better absorbed than quercetin itself. By contrast, glycosides containing a rhamnose moiety could not be absorbed in the small intestine.

[Kalliste]

 

Quercetin glucuronide can apparently get converted back locally into free quercetin aglycone, at inflammation/mitochondrial dysfunction sites, with help from macrophages. I wonder if that is relevant for senescent cells as well?

I had a long post about why upregulate autophagy while doing senolytics was a good idea, but its gone now.
Probably about 6 hours worth of research, links, and typing; just gone...

Isn't it amazing how Google Docs and Gmail are able to save every keystroke..?

IP.Board, this forum software, does have an autosave feature:
http://community.inv...auto-save-post/
But until its enabled my reply has to be:

Yes they are related and it does seem to be a good idea to upregulate autophagy while doing Senolytics.

It seems to be tying into the NAD+ World too:

"...quercetin increases gene expression of the NQO1 gene via an Nrf2 transcription factor mediated pathway(ref)(ref)(ref). Specfically, quercetin enhances the binding of Nrf2 to the NRF-ARE binding site on the NQO1 gene promoter. Quercetin also increases Nrf2-mediated transcriptional activity by up regulation gut e expression of Nrf2 mRNA and Nrf2 protein. Quercetin also reduces the level of Keap1 protein, the binding partner of Nrf2, which prevents Nrf2 nuclear translocation. Quercetin reduces Keap-1in a post-translational mechanism, thereby reducing Nrf2 ubiquitination and proteasomal degradation..."
http://www.longecity...ted-aging-part/

 

 

When writing very long posts it is always prudent to do so in a document and have autosave enabled for every 50 letters or so. This forum is a bit buggy and there is always the occasional computer reboot out of the blue, power outage and so on. Spending 6 hours on a non-saved post seems a bit optimistic. I was burnt like that many times before changing my habits.

[Fafner55]

Luteolin is another flavonoid, similar in structure to quercetin, that is known to induce apoptosis by activating the caspase cascade at similar plasma levels as quercetin.  “Pro-apoptotic effects of the flavonoid luteolin in rat H4IIE cells.” (2005)  ...we analysed DNA strand break formation by luteolin and found a distinct increase of DNA strand breaks after incubation for 3h with 100 microM luteolin, a concentration which induces oligonucleosomal DNA cleavage at 24h. In conclusion, the sequence of events is compatible with the assumption that luteolin triggers the mitochondrial pathway of apoptosis, probably by inducing DNA damage.  

 

Another study of luteolin’s caspase activation to induce apoptosis indicates doses feed to rats.  “Caspase activation and extracellular signal-regulated kinase/Akt inhibition were involved in luteolin-induced apoptosis in Lewis lung carcinoma cells“ (2007) states … In addition,  luteolin significantly inhibited the growth of LLC cells implanted on the flank of mice to 40% and 60% of untreated control group values at 2 mg/kg and 10 mg/kg, respectively.  These doses are 4% to 20% of the quercetin doses used to clear senescent cells in mice.

 

Large doses of luteolin (100 mg / 10 kg body weight) were given to children daily for 26 weeks without major adverse effects.  “An open-label pilot study of a formulation containing the anti-inflammatory flavonoid luteolin and its effects on behavior in children with autism spectrum disorders.” (2013)

Another study supports of luteolin for inducing apoptosis at the same concentration as quercetin.  (Table 1 of http://scholarworks.sjsu.edu/cgi/viewcontent.cgi?article=7788&context=etd_theses).  This dose is 20% of the quercetin doses used to clear senescent cells in mice.  Luteolin is said to have high bioavailability.

Taking an 8000 mg dose of quercetin dihydrate per day for two or three days appears to be a good starting point since there is large individual variability and few side effects.  Experimentation is needed to determine an effective dose.  Unfortunately, I do not currently have metabolic markers or observable measures of the effects of quercetin.  In addition to quercetin dihydrate, a 1600 mg dose of luteolin (20% of quercetin) can also be taken with reasonable safety.

 

 

My Experience with Clearing Senescent Cells

 

2015-03-13 to 15  I took 3200 mg of quercetin twice per day for three days last weekend along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg). Within days after my first treatment, the skin on my arms and body is noticeably smoother and clearer.  Now, three days later, my skin is clearer.  Age spots (raised brown areas) on my arms that I closely watch have noticeably faded, possibly due to the effects of 22 mg of C60-OO I took on 2015-03-15.  I will not take C60 again while this experiment is ongoing.

2015-03-20 Took pictures of my right hand and forearm for reference.

2015-03-22 I took 6400 mg quercetin all at once, along with pterostilbene (100 mg) , icariin (100 mg), resveratrol (1000 mg) and nicotinamide riboside (1000 mg).

2015-03-24  The cells on the backs of my hand appear to be more uniform and smooth.  Age spots on my arms appear slightly smaller but are starting to darken.  It may have been the influence of C60-OO taken on 2015-03-16 that caused them to fade initially.

2015-03-27  I took 8000 mg of quercetin and 1600 mg of luteolin in a single dose.  An hour earlier I took  pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (1000 mg).   

At 3 hours, experienced slight muscle aches and a slight headache.

At 4 hours, I feel normal.

At 6 hours, an area of skin about 0.3 x 0.5 cm on my left hand is red.  Otherwise I look and feel normal.

2015-03-28  The redness on a patch of skin on my left hand has substantially receded.

2015-03-29  The redness on a patch of skin on my left hand is gone.  That area of skin appears different than normal skin; it appears to be a keratosis.

2015-03-29  Took pictures of my right hand and forearm and could not see a change in aged spots compared to 2015-03-20.  

2015-04-01  Icariin causes cells to resist apoptosis and will be eliminated from my senescent cell clearance treatment.  And, because of the side effects associated with the last treatment I am reducing luteolin to 800 mg.  

I took 8000 mg of quercetin, 800 mg of luteolin,  pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (625 mg) in a single dose.  I experienced no side effects.

2015-04-02  I took pterostilbene (100 mg), resveratrol (500 mg) and nicotinamide riboside (625 mg) in the morning, followed by 8000 mg of quercetin and 800 mg of luteolin in the late afternoon.  I experienced no side effects.

2015-04-03  I could not be certain of the effect of NR on this experiment so I stopped taking it.  Today I took only 8000 mg of quercetin and 800 mg of luteolin.  I experienced no side effects.

2015-04-04  Today I took another dose of 8000 mg of quercetin and 800 mg of luteolin along with pterostilbene (100 mg) and resveratrol (500 mg).  I experienced no side effects.

2015-04-05  Took another dose of 8000 mg of quercetin and 800 mg of luteolin along with pterostilbene (100 mg) and resveratrol (500 mg).  Experienced lingering muscle fatigue and soreness in legs after a 2.5 mile walk.

At this point, if quercetin is effective, it should have done it’s work.  I  plan to do nothing further other than wait a few days to see if the raised brown areas on my arms recede.

2015-04-09  Took pictures of my right hand and forearm and could not see a change in aged spots or skin texture from the earlier pictures.

2015-04-10  My 59 year old skin looks the same as I did before beginning this treatment.  If senescent cells have been cleared from my body to any degree, I cannot observe an effect.

 

 

Research suggests that nicotinamide riboside could inhibit the caspase cascade and suppress apoptosis.  My intake of NR could be the reason I did not see an effect from quercetin.  In about a month I will try again, and next time will not to take any other supplement within a week of the senolytic.

 

Mitigation of gamma-radiation induced abasic sites in genomic DNA by dietary nicotinamide supplementation: metabolic up-regulation of NAD(+) biosynthesis.

http://www.ncbi.nlm....pubmed/23891603

 ...A significant increase in liver DNA AP sites and 8-oxo-dG levels with concomitant increase in caspase-3 was observed in CD (control diet) fed and irradiated animals compared to NSD (nicotinamide supplemented diet) fed and irradiated mice. In conclusion present studies show that under γ-radiation stress conditions, dietary nicotinamide supplementation restores DNA excision repair activity via prolonged activation of PARP1 and PARG activities. Present results clearly indicated that hepatic NAD(+) replenishment might be a novel and potent approach to reduce radiation injury.

 

NAD+ acts on mitochondrial SirT3 to prevent axonal caspase activation and axonal degeneration.

http://www.ncbi.nlm....pubmed/23975935

In chronic degenerative syndromes, neuronal death occurs over long periods, during which cells progressively lose their axons and, ultimately, their cell bodies. Although apoptosis is recognized as a key event in neuronal death, the molecular mechanisms involved in CNS axons degeneration are poorly understood. ... We show here that local application of proapoptotic stimuli on the somatodentritic compartment triggers a dying-back pattern involving caspase-dependent axonal degeneration. Using complementary pharmacological and genetic approaches, we further demonstrate that NAD(+) and grape wine polyphenols prevent axonal apoptosis and act via mitochondrial SirT3 activation in axons.

 

Characterization of NAD Uptake in Mammalian Cells

http://www.jbc.org/c...83/10/6367.full

Here we show that various mammalian cell types are able to transport picomolar concentrations of NAD across the plasma membrane from the extracellular medium. When intracellular NAD(P) levels were reduced by treatment with the NAD biosynthesis inhibitor FK866 (APO866, (E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-(pyridin-3-yl)acrylamide (25)), application of NAD extracellularly was able to partially replenish these levels. Furthermore, cells could be rescued from FK866-induced NAD depletion, autophagy, and cell death by addition of NAD to the culture medium. These data, taken together, suggest that NAD uptake can participate in the homeostasis of cellular NAD levels.

[pone11]

 

Research suggests that nicotinamide riboside could inhibit the caspase cascade and suppress apoptosis.  My intake of NR could be the reason I did not see an effect from quercetin.  In about a month I will try again, and next time will not to take any other supplement within a week of the senolytic.

 

 

For that matter, stop all antioxidants maybe for a week prior to the senolytic therapy.   You probably don't want to help the cell survive stress.

[niner]

Also, Judith talked about something that seemed troubling to me in regards to potential side effects of senolytics. Senescent cells may play an important and beneficial role in healing and suppressing senescent cells can apparently impair healing in mice

 

I finally got around to watching that talk by Dr. Campisi.  It's a good talk- thanks for posting it.  I don't think that wound healing will be a problem, since it sounded to me like the senescent cells that were involved in wound healing were transient.  In other words, they came and went.  Did I hear that right?  Like some cells at the injury site senesce, contribute to healing, but then die apoptotically?  If that's the case, then periodic (e.g. yearly) removal of persistent senescent cells shouldn't have any bearing on wound healing.

[Fafner55]

The Achilles’ Heel of Senescent Cells: From Transcriptome to Senolytic Drugs reports quercetin to be effective in inducing apoptosis in senescent HUVECs but not preadipose cells.

 

After taking large doses of quercetin, I did not observe an effect to my skin. An explanation could be that quercetin is not effective in inducing apoptosis in senescent dermal fibroblasts, since dermal fibroblasts as known to be highly resistant to apoptosis, as are preadipose and adipose cells, whereas senescent HUVECs are significantly more apoptosis prone. Apoptosis Resistance in Senescent cells, Cellular Senescence: From Growth Arrest to Immunogenic Conversion

 

It may very well be that the combination of quercetin + dasatinib will be needed to clear the body of a wide range of senescent cells.

[Logic]

Other Quercetin effects that may of of interest here:

http://www.longecity...ndpost&p=596717

http://www.longecity...ndpost&p=702748

 

(I haven't yet followed the links provided in the above and related posts)

[corb]

The Achilles’ Heel of Senescent Cells: From Transcriptome to Senolytic Drugs reports quercetin to be effective in inducing apoptosis in senescent HUVECs but not preadipose cells.

 

After taking large doses of quercetin, I did not observe an effect to my skin. An explanation could be that quercetin is not effective in inducing apoptosis in senescent dermal fibroblasts, since dermal fibroblasts as known to be highly resistant to apoptosis, as are preadipose and adipose cells, whereas senescent HUVECs are significantly more apoptosis prone. Apoptosis Resistance in Senescent cells, Cellular Senescence: From Growth Arrest to Immunogenic Conversion

 

It may very well be that the combination of quercetin + dasatinib will be needed to clear the body of a wide range of senescent cells.

 

I've been meaning to ask - why do you think quercetin can do anything for your skin? The original paper posted here didn't talk about dermal cells.

Both quercetin and dasatinib were shown to work with efficiency only on a handful of cell types.
 

[niner]

I've been meaning to ask - why do you think quercetin can do anything for your skin? The original paper posted here didn't talk about dermal cells.

Both quercetin and dasatinib were shown to work with efficiency only on a handful of cell types.

 

The hope would be that most or all cells would share the same apoptotic pathways that are not functioning properly in senescent cells, and that the D/Q combination would work to a greater or lesser extent in all of them.  That remains to be shown, of course.  Were there any cell types that the senolytics guys looked at that were resistant to the combination?
 

[Fafner55]

 

The Achilles’ Heel of Senescent Cells: From Transcriptome to Senolytic Drugs reports quercetin to be effective in inducing apoptosis in senescent HUVECs but not preadipose cells.

 

After taking large doses of quercetin, I did not observe an effect to my skin. An explanation could be that quercetin is not effective in inducing apoptosis in senescent dermal fibroblasts, since dermal fibroblasts as known to be highly resistant to apoptosis, as are preadipose and adipose cells, whereas senescent HUVECs are significantly more apoptosis prone. Apoptosis Resistance in Senescent cells, Cellular Senescence: From Growth Arrest to Immunogenic Conversion

 

It may very well be that the combination of quercetin + dasatinib will be needed to clear the body of a wide range of senescent cells.

 

I've been meaning to ask - why do you think quercetin can do anything for your skin? The original paper posted here didn't talk about dermal cells.

Both quercetin and dasatinib were shown to work with efficiency only on a handful of cell types.
 

 

 

The authors only discussed a limited number of cell types.  They reported that dasatinib was effective in preadipose cells while quercetin was effective against senescent human endothelial cells.   They also stated that quercetin was effective in mouse BM-MSCs.  There was no mention any cell type that was resistant to the combination of D+Q.

 

Dermal fibroblasts, which are known to be resistant to apoptosis, were not discussed.  I monitored my skin on the off hope that quercetin might be effect and because skin is one of the few things I can observe.

[corb]

 

I've been meaning to ask - why do you think quercetin can do anything for your skin? The original paper posted here didn't talk about dermal cells.

Both quercetin and dasatinib were shown to work with efficiency only on a handful of cell types.

 

The hope would be that most or all cells would share the same apoptotic pathways that are not functioning properly in senescent cells, and that the D/Q combination would work to a greater or lesser extent in all of them.  That remains to be shown, of course.  Were there any cell types that the senolytics guys looked at that were resistant to the combination?
 

 

 

From the original paper :

 

 

Interfering with expression of the ephrin dependence receptor ligands, EFNB1 or EFNB3, induced selective loss of senescent cells. Ephrin receptors are the largest family of  receptor tyrosine kinases (Boyd et al. 2014). Together with ephrin ligands, these receptors coordinate tissue and organ patterning, cell positioning, and cell survival during development and tissue turnover in a cell type-specific manner (Xi et al.2012).

 

The paper led me to believe they only looked at a small number of cell types because they found drugs that act on only those cells through their specific ligands. Further into the paper they say:

 

 

Other dependence receptor networks, which promote apoptosis unless they are constrained from doing so by presence of ligands, might be particularly informative to study, especially to develop cell type-, tissue-, and disease-specific senolytic agents. These receptors include the insulin, IGF-1, androgen and nerve growth factor receptors, among others (Delloye-Bourgeois et al. 2009; Goldschneider & Mehlen 2010

 

So I think their implication is Dasatinib and Quercetin don't work on a common pathway.

 

I could be wrong. The paper isn't organized well.

[Fafner55]

There is reason for optimism that the combination D+Q will address enough cell types to make a meaningful difference in heath.

 

From Figures 10 and 11 of the supplemental material available at http://onlinelibrary....12344/suppinfo, many age score factors showed improvement as I summarize below

 

Improved

Loss of grip strength

Hind Limb Paralysis

Gait Disorders

Body Condition = an assessment of the animal's weight for age, and its relative proportions of muscle and fat.

Ataxia = the loss of full control of bodily movements.

Cataracts (supplemental figure 10)

 

Did Not Improve

Tremor

Kyphosis = excessive outward curvature of the spine, causing hunching of the back.

Dystonia = a state of abnormal muscle tone resulting in muscular spasm and abnormal posture.

 

The treated mice in Figure 10 generally look healthier than the untreated mice. And, the irradiated mouse in Figure 5 of the original paper looked in good shape.

 

Edited by Fafner55, 18 April 2015 - 04:06 PM.

[nowayout]

As for what we should expect for skin:

 

From figure 11 it appears that some of the D+Q treated animals actually had worse coat condition (yellow bars) than vehicle-treated animals. 

[niner]

The improvements that were seen in the mice covered both muscle and nerve tissues, so it appears that it's working in multiple cell types beyond the lines tested in vitro.  It's not helping everything, but some of the problems (kyphosis, for example) might have negligible input from senescent cells.  I think it's pretty impressive to see such widespread improvement from a zeroeth generation treatment, and only one brief treatment period at that.   As far as the animals coat condition, unless there was an adverse effect from either of the compounds, I don't see why the loss of some senescent cells would be bad for the coat.  If the coat effect is real, then we'll have to wait and see if it's a class effect of senolytics, or if it's specific to these agents.

[mike_nyc]

Thorne has a new quercetin phytosome.

 

https://shop.thorne....cetin-phytosome