1079 replies to this topic

[Martini]

In the first study, it was shown that Tween 80/limonene/nisin with little agitation produced droplets of just 20nm, but that Tween 80 and limonene without nisin also made a nano emulsion.  There is a discussion also of how it gets mixed which I haven't digested yet.  But it seems that with the Tween 80 and limonene mix it may not be a problem since it apparently mixes easily without little agitation.  "In addition, the presence of some bioactive compounds may have minimal effect on particle size. A small increase in particle diameter was observed when nisin was added to a nanoemulsion made with 4% oil (D‐limonene) and 6% surfactant."  So, only a small increase with the added nisin.  Tween 80 and 40 are the best surfactants for particle diameter as Figure 6 shows.  Hence, 90% water, 4% D-limonene, and 6% Tween 40 as a formula for a nano emulsion of fisetin is my theory.

 

I think you need to know the solubility of Fisetin in the Tween 40 + D-limonene mixture (the self-microemulsifying drug delivery system) to achieve Fisetin loaded microemulsions. Do you have idea about it?
 

[manofsan]

So you guys mention it's a terpene. By coincidence, I like to take Gotu Kola as a tea, and that's supposed to contain some important terpenes which are good for the blood vessels of the brain. I wonder if adding some D-Limonene into my Gotu Kola tea would increase bioavailability of the Gota Kola terpenes as well?

[LarryG]

FYI, here is a study testing quercetin transdermal by dissolving in various solvents but not DMSO.  While they thought quercetin had promise applied to the skin for the skin per se, however, "In summary, quercetin even when formulated in a lipid nanoparticle vector shows no evidence for transdermal delivery on human skin. Keeping in mind that quercetin as a molecule is a paradigmatic model for a lipophilic drug (octanol−water partition coefficient log P = 1.82) [73] with 5 polar hydroxyl heads and very low water solubility [81], thus quercetin is not the perfect drug candidate for a transdermal delivery system. Quercetin local skin deposition is more valuable than performing a transdermal delivery through skin. Quercetin envisaged dermal applications described above (section 2) are all of local interest and the absence of a systemic absorption is desirable. Nanodosage forms were able to increase quercetin skin retention via their occlusive effect and higher surface area. Transdermal delivery for quercetin nanodosage forms was not achieved without the help of penetration enhancers."  So, given the closeness of fisetin to quercetin in structure, the same may be true of fisetin.

 

 

(Note: probably taken from this source: https://www.scienced...39641116304714 -MaxWatt)

 

 

Edited by maxwatt, 19 March 2019 - 03:13 AM.

[Woody42]

I am considering taking 1,000 mg a day for 5 days but in view of it's short half life I wonder

if I should take it in decided doses or all at once. 

[maxwatt]

If the pharmokinetics are similar to resveratrol there is a maximum dose beyond which taking more does not increase blood levels as much as if the curve were linear.   My guess is that the knee of the curve occurs around 1 gram, where you have diminishing returns.  Beyond 4 of 5 grams there is unlikely to be any benefit.

 

Half life of resveratrol and likely other polyphenols is likely to be about 2 hours, with hepatic recirculation resulting in a secondary blood level peak at about 8 hours, plus or minus an hour or two.

 

This is guesswork, assuming the metabolization to to be similar to resveratrol.  If I have the time I'll see if I can find which enzymes the body uses to process it.

 

Also adding methyl groups or acetylating the molecule are likely to increase effectiveness.  Somewhere a lab is probably working on it.

[OP2040]

I've settled into taking just 200 mg/day and maybe will take a high dose for a short period each year.  It's clear by this thread that we all tend to overthink these things.  At some point you just have to trust the science.  And there is a lot of good science surrounding Fisetin.  We shouldn't expect miracles from senolytics, it is just one more stop on the line toward our goal.  It will probably not allow you to live one single day longer, but it may very well prevent an early death, thus allowing you to be alive when more powerful tools arrive.

 

All of this self-reporting and recipe-tweaking is a time-waster if you ask me.  There is only one data point that we need.  If we wanted to we could determine if the supplement Fisetin is actually removing our senescent cells.  The tools for this are accessible and manageable, but no one wants to do it, they just want endless guessing.  If we could determine that baseline level of effectiveness, then all the debating goes out the window and we can take it and move onto greener pastures.

 

The thing that people don't seem to get, whether here or life in general, is the concept of opportunity cost.  Every second that's spend on something that is not a complete cure for aging, is a second taken away from research and efforts toward that cure.  Time is far more precious than money, and that's why those of us that learned economics in university remember that much of it is wrapped up in the "time-value" concept of money.  So lets get on with it.  I'll ask one last time, does anyone want to pursue an actual test of whether Fisetin eliminates senescent cells with me?  If not, I'll see you in the cellular reprogramming threads.

[Rays]

does anyone want to pursue an actual test of whether Fisetin eliminates senescent cells with me? 

 

I'm interested. How would you go about it and what would it cost?

[OP2040]

Hi Rays,

I would have to do look into it again.  From what I remember it is not straightforward, as in a simple test.  There are tests for senescent cells, b-gal being the most used.  They are rather pricey if it's just one person buying, because it is assumed a lab is buying at bulk.  For example here is a really pricey one, but if you look it actually has 250 tests included.

 

https://www.abcam.co...it-ab65351.html

 

Aside from price, the other issue is tissue specificity.  I assume it will test for cells in whatever tissue you can get your hands on.  So for an in vivo test, we are limited to probably skin depending on how daring you are lol.  So it's a similar issue to telomere testing where you are almost always getting LTL and wondering how representative that is.  I am thinking that the science of senescent cells is advanced enough that there should be some research out there both for typical senescent cell burden by age and by various tissue types. 

 

Finally, the one thing that could derail the idea is that a microscope is needed.  I read this as a standard microscope, which I can access at a lab at my workplace, or just buy used.  But if it's anything more that may be a show-stopper

 

So it is plausible to get a baseline, intervene and then see what happens. I've been taking Fisetin for some time, so I may just be interested in finding out my baseline, or doing a high-dose, and then see if that makes a difference.

 

So there are a few hurdles, but it's extremely valuable information if you ask me.  Possibly more valuable than a telomere test at this point.

 

There may be other options out there, like testing for SASP level, but this seemed the most straightforward. 

[Rays]

€415 for 250 tests makes €1.66 per test. That's not expensive if enough people take part.

But as you said, a microscope is needed. It needs 200x magnification, I saw in the booklet:

 

"Observe the cells under a microscope for development of blue color (200X total magnification)."

 

Additional materials required:

 Pipettes and pipette tips

 PBS Solution

 DMSO or DMF (N-N-dimethylformamide)

 12 well plate

 

Could you do the tests for several subjects, if they send blood to you?

 

I have not yet taken fisetin, so I would be a good "before and after" test subject.

 

[OP2040]

I didn't see the DMSO, so not sure how hard that is to get.  There are other sites that offer pre-made solutions which may help with that.  A group buy would be really hard to coordinate, what with sending tissue samples through the mail.  So I'm not sure that's the route we'd want to go.  Another thing we didn't think of is the analysis.  I assume that you can just tell that they've decreased, but who knows if there is software that studies use to analyze assays, since counting seems rather primitive.  A lot to take into consideration but I am glad to see some interest being generated.

[OP2040]

I think the first step is to find a study with baselines for senescent cell burden by age for various tissue types.  I have to believe there's a study out there on it somewhere.  I wouldn't want to use mice as a proxy either.

[Martini]

I've settled into taking just 200 mg/day and maybe will take a high dose for a short period each year.  It's clear by this thread that we all tend to overthink these things.  At some point you just have to trust the science.  And there is a lot of good science surrounding Fisetin.  We shouldn't expect miracles from senolytics, it is just one more stop on the line toward our goal.  It will probably not allow you to live one single day longer, but it may very well prevent an early death, thus allowing you to be alive when more powerful tools arrive.

 

All of this self-reporting and recipe-tweaking is a time-waster if you ask me.  There is only one data point that we need.  If we wanted to we could determine if the supplement Fisetin is actually removing our senescent cells.  The tools for this are accessible and manageable, but no one wants to do it, they just want endless guessing.  If we could determine that baseline level of effectiveness, then all the debating goes out the window and we can take it and move onto greener pastures.

 

The thing that people don't seem to get, whether here or life in general, is the concept of opportunity cost.  Every second that's spend on something that is not a complete cure for aging, is a second taken away from research and efforts toward that cure.  Time is far more precious than money, and that's why those of us that learned economics in university remember that much of it is wrapped up in the "time-value" concept of money.  So lets get on with it.  I'll ask one last time, does anyone want to pursue an actual test of whether Fisetin eliminates senescent cells with me?  If not, I'll see you in the cellular reprogramming threads.

 

Fisetin selectively reduces viability of senescent human HUVECs (endothels), but not IMR90 (fibroblast) cells or primary human preadipocytes. So it is only working selectivley on some cell types. If you search the wrong place, you will come to a wrong conclusion. So we know already that Fisetin is not removing our senescent cells in general!
 

[Krell]

I think you need to know the solubility of Fisetin in the Tween 40 + D-limonene mixture (the self-microemulsifying drug delivery system) to achieve Fisetin loaded microemulsions. Do you have idea about it?
 

 

FWIW here are my latest thoughts/interpretations regarding the nano emulsion technique for increasing the bioavailability of fisetin

 

So far, several of us have taken a gram or two of fisetin powder mixed in evoo or alcohol/water mixture, with inconclusive results.

 

I have tried testing the solubility of fisetin in eevo or vodka by mixing ~30 grams of fluid (shot glass) with 100mg fisetin (one capsule), and it seems only partly soluble if I wait 24 hours for the suspended solids in the mixtures to settle out. I have not tried extended stirring with a mechanical stirrer.

 

The nano emulsifying approach requires dissolving fisetin in an oil (D-limonene) and then breaking the oil into nanometer droplets by an pouring the oil into the emulsifier (water-tween 80 mixture) and stirring.

 

The nano-emulsion seems to increase the bioavailability of the dissolved substance because of the nanometer droplet high surface to volume ratio and other less well known effects as it passes through the gut.

 

I have some d-limonine on order and I will try some experiments to dissolve fisetin in it.  One poster found that the solubility of quercetin was reported as ~15mg/ml, and he guessed that fisetin might be similar.  I plan to first run solubility tests to see if the estimate is in the ball park.

 

Assuming 15mg/ml solubility and my Mayo Clinic suggested fisetin dose (83kg*20mg/kg=1660mg) then I would need 1660mg/15mg/ml=111ml of D-limonene to dissolve my dose.

 

Then if I use the suggested nano-emulsion proportions of 90% water, 4% D-limonene, 6% Tween 80, I will need 27.4ml/% or about 2500ml=2.5 liters of solution to drink. That is about 6 large water glasses, so I might need some hours to drink it.  It would certainly be advisable to do test runs on less volume and with fewer ingredients to determine if one is sensitive to any of the ingredients.

 

Any comments?
 

[manofsan]

From what I see, a lot of Fisetin is extracted from strawberries. And as we know, fermentation of foods (including strawberries) can concentrate some of their nutrients in the fermentation product (exactly which nutrients depends upon the type of fermentation bacteria used.)

 

So can there be a way to produce concentrated Fisetin in a fermented product, for healthful consumption?

Edited by manofsan, 19 March 2019 - 09:10 PM.

[Martini]

If the pharmokinetics are similar to resveratrol there is a maximum dose beyond which taking more does not increase blood levels as much as if the curve were linear.   My guess is that the knee of the curve occurs around 1 gram, where you have diminishing returns.  Beyond 4 of 5 grams there is unlikely to be any benefit.

 

Half life of resveratrol and likely other polyphenols is likely to be about 2 hours, with hepatic recirculation resulting in a secondary blood level peak at about 8 hours, plus or minus an hour or two.

 

This is guesswork, assuming the metabolization to to be similar to resveratrol.  If I have the time I'll see if I can find which enzymes the body uses to process it.

 

Also adding methyl groups or acetylating the molecule are likely to increase effectiveness.  Somewhere a lab is probably working on it.

 

I was looking ino it in the past. From the CYP450 familiy it is CYP1A2, CYP2C9, CYP2C19 and CYP2E1. I think a critical point is the toxic level around 60 µMol/ml in vitro. As there are no studies for the toxic levels of the metabolites one should be prudent.

[Martini]

FWIW here are my latest thoughts/interpretations regarding the nano emulsion technique for increasing the bioavailability of fisetin

 

So far, several of us have taken a gram or two of fisetin powder mixed in evoo or alcohol/water mixture, with inconclusive results.

 

I have tried testing the solubility of fisetin in eevo or vodka by mixing ~30 grams of fluid (shot glass) with 100mg fisetin (one capsule), and it seems only partly soluble if I wait 24 hours for the suspended solids in the mixtures to settle out. I have not tried extended stirring with a mechanical stirrer.

 

The nano emulsifying approach requires dissolving fisetin in an oil (D-limonene) and then breaking the oil into nanometer droplets by an pouring the oil into the emulsifier (water-tween 80 mixture) and stirring.

 

The nano-emulsion seems to increase the bioavailability of the dissolved substance because of the nanometer droplet high surface to volume ratio and other less well known effects as it passes through the gut.

 

I have some d-limonine on order and I will try some experiments to dissolve fisetin in it.  One poster found that the solubility of quercetin was reported as ~15mg/ml, and he guessed that fisetin might be similar.  I plan to first run solubility tests to see if the estimate is in the ball park.

 

Assuming 15mg/ml solubility and my Mayo Clinic suggested fisetin dose (83kg*20mg/kg=1660mg) then I would need 1660mg/15mg/ml=111ml of D-limonene to dissolve my dose.

 

Then if I use the suggested nano-emulsion proportions of 90% water, 4% D-limonene, 6% Tween 80, I will need 27.4ml/% or about 2500ml=2.5 liters of solution to drink. That is about 6 large water glasses, so I might need some hours to drink it.  It would certainly be advisable to do test runs on less volume and with fewer ingredients to determine if one is sensitive to any of the ingredients.

 

Any comments?
 

 

My understanding is that you first mix the D-limonene with the tween 80 and Fisetin. After mixing you can titrate the water during mixing. This is why I was asking for the solubility of the D-limonene - tween 80 mix with Fisetin. The method is described in DOI: 10.1208/s12249-017-0798-x. In general you need to be prudent with the dosage of a much higher bioavailable Fisetin due to the toxic effect around 60 µmol/ml.
 

[OP2040]

Fisetin selectively reduces viability of senescent human HUVECs (endothels), but not IMR90 (fibroblast) cells or primary human preadipocytes. So it is only working selectivley on some cell types. If you search the wrong place, you will come to a wrong conclusion. So we know already that Fisetin is not removing our senescent cells in general!
 

 

Although it is often painful, this is why a reference or quote is important for any of these discussions.  The very study this mammoth thread is based on showed Fisetin to be very effective at removing senescent cells from various tissues, including fibroblasts.  Here is one of the relevant quotes from the study:

 

 

A panel of flavonoid polyphenols was screened for senolytic activity using senescent murine and human fibroblasts, driven by oxidative and genotoxic stress, respectively. The top senotherapeutic flavonoid was tested in mice modeling a progeroid syndrome carrying a p16INK4a-luciferase reporter and aged wild-type mice to determine the effects of fisetin on senescence markers, age-related histopathology, disease markers, health span and lifespan. Human adipose tissue explants were used to determine if results translated.

Of the 10 flavonoids tested, fisetin was the most potent senolytic. Acute or intermittent treatment of progeroid and old mice with fisetin reduced senescence markers in multiple tissues, consistent with a hit-and-run senolytic mechanism. Fisetin reduced senescence in a subset of cells in murine and human adipose tissue, demonstrating cell-type specificity. Administration of fisetin to wild-type mice late in life restored tissue homeostasis, reduced age-related pathology, and extended median and maximum lifespan.

 

It is certainly reasonable to assume it doesn't work in every single possible tissue/cell.  But it doesn't have to in order to have body-wide effects.  In the cited study it worked in every tissue type studied, and it was a very well done study. Please reference at least the title of your study.

[Martini]

Although it is often painful, this is why a reference or quote is important for any of these discussions.  The very study this mammoth thread is based on showed Fisetin to be very effective at removing senescent cells from various tissues, including fibroblasts.  Here is one of the relevant quotes from the study:

 

 

It is certainly reasonable to assume it doesn't work in every single possible tissue/cell.  But it doesn't have to in order to have body-wide effects.  In the cited study it worked in every tissue type studied, and it was a very well done study. Please reference at least the title of your study.

 

sorry, you are right. Here is the link https://doi.org/10.18632/aging.101202

Obviously, there is a difference in the effectiveness for humans and mice. Another good overview for senolytics is also  https://doi.org/10.1155/2018/4159013

[OP2040]

Thanks, hadn't seen that study yet. 

 

I just read a study that claims, using some empirical testing and a lot of math, that senescent cell dynamics can completely explain the Gompertz mortality law.  If that is even half true, it's a really big deal.

 

The other interesting thing from the article is that the production of senescent cells goes up with age alongside the decrease in the inability to remove them, which I'm sure we already knew, but rarely does it fit into the equation when we discuss senolytics.  This would mean that senolytics should be very powerful, but only half of the solution.  We also need to prevent the increased rate of production of senescent cells, presumably with telomeres or epigenetics.  It seems clear from this that the next experimental step in mice should be clearance of senescent cells alongside telomere or epigenetic rejuvenation.  According to this study, that would flatten the Gompertz curve like no other intervention has come close to doing yet.

 

https://www.biorxiv....0.1101/470500v2

 

I'm really struggling to find a study that shows any kind of measurement of senescent cell burden by age in humans.  There's plenty out there on mice, as usual.  If anyone finds one in humans, please post.  We need that baseline to proceed.

[Woody42]

I took  2 doses of 500mg of fisetin yesterday disolved in a little dmso and water.

I feel almost like I have a mild hangover without drinking.  But speaking of alchohol

I saw a listing that fisetin is soluble in ethyl alcohol  5mg/ml  and  30mg/ml in dmso.

So I wonder if this feeling like I have a hangover is from the 20 ml of dmso I used 

or if it could be senescent cells dying in my 68 year old body.

   

 

[manofsan]

This would mean that senolytics should be very powerful, but only half of the solution.  We also need to prevent the increased rate of production of senescent cells, presumably with telomeres or epigenetics.  It seems clear from this that the next experimental step in mice should be clearance of senescent cells alongside telomere or epigenetic rejuvenation.  According to this study, that would flatten the Gompertz curve like no other intervention has come close to doing yet.

 

In a way, aren't we really just fighting Entropy?

 

Anything informational can degrade - genetic, epigenetic, etc.

 

We already know about Telomerase, but how do we fix the epigenetic side of things?

[OP2040]

In a way, aren't we really just fighting Entropy?

 

Anything informational can degrade - genetic, epigenetic, etc.

 

We already know about Telomerase, but how do we fix the epigenetic side of things?

 

I'm just one man with one opinion, but I am adamantly opposed to entropy (in itself) being the cause of aging.  There is a lot of logic and evidence that can lead us to that conclusion.  Starting with the basics, living things are open systems, so right from the beginning we can say that it can't be the cause.  The reason it looks like entropy is applicable to living things is because evolution is biased toward reproduction and therefore biased against individual survival.  There are negligibly senescent and regenerative animals, but you always find them in non-reproductive animals (jellyfish) or animals with alternative reproductive strategies (naked mole rats). 

 

It may very well be that damage is what kills us, but damage in an open system need not mean the demise of the overall system, as it can be balanced by energy intake.  The fact that calorie restriction boosts repair and lifespan really underlines the point that the body could easily repair itself if it had the instructions to do so. 

 

The epigenetic side of things should be able to be fixed by cellular reprogramming.  For a time, it was thought that the epigenetic clock was a whole separate and new mechanism because it didn't return to a youthful state with cellular reprogramming.  And that is true for telomere restoration.  But it is not true for cellular reprogramming.  As we can see from the evidence coming in, reprogramming returns the epigenetic state to youth, and it also returns telomeres to a youthful level.  It's my opinion that epigenetic reprogramming will cover almost all of our bases when it comes to rejuvenation.  And aside from youthfulness, I think we will have specific recipes for specific organs, like say the eye, or the retina.  The recipe will include non-OSKM reprogramming factors that tilt gene expression toward regeneration as it does in adult regenerative animals. 

 

This is the way forward, but I can't deny the appeal of focusing on specific damages, and I do it myself.  It is very tempting to want to purge your body of all the baddies (oxidized cholesterol, amyloid, etc.)  but the evidence is thin that this does much of anything in a systemic environment that is bent on allowing destruction to run rampant.   I don't know what others are reading, but every instance of true regenerative repair that I've seen has involved reprogramming. 

[manofsan]

Yeah, I'll agree with you that while the total Entropy of the universe as a whole has to increase, the entropy of a system (eg. a human being or human beings) can be made to decrease, even while the total Entropy of the universe still increases as required.

 

But in order to accomplish this, we will ultimately have to rely upon systems that are far more isentropic than just mere biochemistry. We'd have to make use of our other man-made technologies to do that.

[OP2040]

The universe is for all practical purposes infinite from our perspective.  Lets just conquer our own biology first, then we can start worrying about various other existential threats.  I do find it interesting that no less a luminary than Leonard Hayflick tried to argue that senescence is inevitable.....because entropy, and even dared to write a paper defending that.  To me, this is just sloppy thinking.  Yes, if nothing changes and we take the current state of affairs as eternal. then the body fails because entropic forces prevail for most species.  It's sloppy thinking in the same way Malthus was shown to be embarrassingly wrong about demographics.  This is what happens when one looks at a system in isolation.  Hayflick is actually making the same exact mistake that Malthus made.  Both of them are trying to isolate a system to examine it, and deliberately ignoring a host of "external" variables in order to simplify and explain.   In the case of both, it is ultimately human will and consciousness that is being ignored.  We had our demographic transition.  Now we will have a biomedical one. 

[manofsan]

I took  2 doses of 500mg of fisetin yesterday disolved in a little dmso and water.

I feel almost like I have a mild hangover without drinking.  But speaking of alchohol

I saw a listing that fisetin is soluble in ethyl alcohol  5mg/ml  and  30mg/ml in dmso.

So I wonder if this feeling like I have a hangover is from the 20 ml of dmso I used 

or if it could be senescent cells dying in my 68 year old body.

 

When I've taken Fisetin, I later get hyperactivity/energy combined with hunger pangs (I've nearly always taken it during exercise)

 

The other day, I tried using Piperlongumine - on its own, because I didn't want to stack things the first time out. I also later got hyperactivity/energy combined with hunger pangs.

 

Today, I tried both the Fisetin & Piperlongumine together during my workout, and felt a totally different reaction afterwards. I felt extremely and uncharacteristically fatigued and lacking in energy, for a period of a few hours following the end of my workout. Then after that passed, I had some extra energy, felt myself tapping my feet, even while feeling hunger pangs (I'd already had a big lunch after my workout).

 

Has anyone else tried both Fisetin & Piperlongumine together, and experienced this latter kind of reaction? Is the kind of reaction I experienced a sign of something good or bad?

[Woody42]

Fore the last 4 days I have been using 1000 mg of fisetin, 250 mg turmeric and 5 mg piperine. 

I mixed the fisetin and turmeric in 5ml of DMSO and then diluted it with water.  I felt a little fatiuged

and almost like I had a hangover plus I had a mild headache. It seems that I didn't feel as much

reaction the 3rd day and almost no reaction yesterday so I may end my experiment today. I did 

go to the gym yesterday and found it a little more fatiguing than it usually is for me and didn't get

any extra energy later on like you did. 

 

[Oakman]

When I've taken Fisetin, I later get hyperactivity/energy combined with hunger pangs (I've nearly always taken it during exercise)

 

The other day, I tried using Piperlongumine - on its own, because I didn't want to stack things the first time out. I also later got hyperactivity/energy combined with hunger pangs.

 

Today, I tried both the Fisetin & Piperlongumine together during my workout, and felt a totally different reaction afterwards. I felt extremely and uncharacteristically fatigued and lacking in energy, for a period of a few hours following the end of my workout. Then after that passed, I had some extra energy, felt myself tapping my feet, even while feeling hunger pangs (I'd already had a big lunch after my workout).

 

Has anyone else tried both Fisetin & Piperlongumine together, and experienced this latter kind of reaction? Is the kind of reaction I experienced a sign of something good or bad?

 

Yes, I have. For my last senescence regimen about 2 weeks ago I used Piper Longum 2 grams, .8 grams Fistein, .5 grams Resveratrol, .6 grams curcumin - for 2 days running in a divided am / noon / pm dosing. This was along with other daily components that I started a day earlier (for 3 days total) of - Vit C 400 mgs, zinc 22 mgs, ginger 1.75 grams, reshi 2 grams, ashwagandha 1.75 grams, betaine hcl 1.35 grams, watercress 400 mgs. 

 

The 1st time I tried this (end 12/2018) I also did pseudo-fasting of ~800 calories/day and I felt pretty awful, fatigued and not right. This last time I ate normally, and all was fine, no strange or ill feelings. So either the fasting didn't sit well with me, or the process purged less and so less upset....it's hard to tell. One thing I felt was that just waiting 1 month between sessions was not enough. I felt that things were gradually improving after the first session last year, but were not finished end Jan in order to try again. 

[manofsan]

Fore the last 4 days I have been using 1000 mg of fisetin, 250 mg turmeric and 5 mg piperine. 

I mixed the fisetin and turmeric in 5ml of DMSO and then diluted it with water.  I felt a little fatiuged

and almost like I had a hangover plus I had a mild headache. It seems that I didn't feel as much

reaction the 3rd day and almost no reaction yesterday so I may end my experiment today. I did 

go to the gym yesterday and found it a little more fatiguing than it usually is for me and didn't get

any extra energy later on like you did. 

 

 

Hiya - take the stuff during your cardio workout, rather than before - take it at the midpoint of your workout. They say a good cardio workout should be no less than 1 hour at the gym, and shouldn't have to go beyond 2 hrs - although in this case where you're taking these senolytics, I'd consider going up to 3 hours total of vigorous cardio exercise. Bring a protein shake with you,  to have it soon after your workout is over.

 

 

Yes, I have. For my last senescence regimen about 2 weeks ago I used Piper Longum 2 grams, .8 grams Fistein, .5 grams Resveratrol, .6 grams curcumin - for 2 days running in a divided am / noon / pm dosing. This was along with other daily components that I started a day earlier (for 3 days total) of - Vit C 400 mgs, zinc 22 mgs, ginger 1.75 grams, reshi 2 grams, ashwagandha 1.75 grams, betaine hcl 1.35 grams, watercress 400 mgs. 

 

The 1st time I tried this (end 12/2018) I also did pseudo-fasting of ~800 calories/day and I felt pretty awful, fatigued and not right. This last time I ate normally, and all was fine, no strange or ill feelings. So either the fasting didn't sit well with me, or the process purged less and so less upset....it's hard to tell. One thing I felt was that just waiting 1 month between sessions was not enough. I felt that things were gradually improving after the first session last year, but were not finished end Jan in order to try again. 

 

So you seem to do it at the lower frequency, once every few months as a megadose. Hmm, I see a lot of people saying similar things, but I'm greedy to get as many improvements as I can upfront, rather than waiting it out. My style right now is to keep taking a couple of hundred mg of the Fisetin & Piperlongumine while going to the gym daily. Once my benefits plateau, I'll stagger it out to every few months as a megadose .

[smithx]

The big issues with the test you linked are:

• It is measuring enzyme activity, so it works only with live cells. That means the sampling has to take place at the lab where the test will be conducted.
• The live cells which would be used are not a blood sample. This test works with tissue culture cells adhered to a surface, such as fibroblasts, or with tissue sections. In our case, we can't use tissue culture so what would be tested is a skin section: in other words, cutting out a piece of live skin.
• Once the skin section is performed, in order to get consistent results, we have to be sure we are always looking at the same layer of skin. The external surface won't work, because the outer layer is mostly dead cells. So we have to turn the skin around and shave it til we have an even layer exposed that's the same in all samples. This is probably not something that anyone untrained should be doing.

 

Other notes:

• You do need some additional equipment, namely an incubator and a microscope.
• To determine how many senescent cells per unit area, you really want a microscope with a grid overlay, so you can count the number of senescent cells per grid area.
• The component left out of the kit is this: https://www.thebalan...d-saline-375492

But these points are moot because skin sections are really not the sort of thing you should be doing at home, or would be likely to do in a repeatable way.

 

 

For example here is a really pricey one, but if you look it actually has 250 tests included.

 

https://www.abcam.co...it-ab65351.html

 

 

 

Edited by smithx, 25 March 2019 - 07:01 AM.

[OP2040]

Although it is often painful, this is why a reference or quote is important for any of these discussions.  The very study this mammoth thread is based on showed Fisetin to be very effective at removing senescent cells from various tissues, including fibroblasts.  Here is one of the relevant quotes from the study:

 

 

It is certainly reasonable to assume it doesn't work in every single possible tissue/cell.  But it doesn't have to in order to have body-wide effects.  In the cited study it worked in every tissue type studied, and it was a very well done study. Please reference at least the title of your study.

 

 

sorry, you are right. Here is the link https://doi.org/10.18632/aging.101202

Obviously, there is a difference in the effectiveness for humans and mice. Another good overview for senolytics is also  https://doi.org/10.1155/2018/4159013

 

Turns out I may have been right about Fibroblasts, but dead wrong in the sense that Fisetin does miss some other fairly important senescent cells.  This study just out today:

 

https://onlinelibrar...111/acel.12950

"Targeting Senescent Cells Enhances Adipogenesis and Metabolic Function in Old Age"

 

 

We employed the combination of D plus Q (D + Q) in our studies for the following reasons. (a) In our hands, no senolytic investigated thus far targets all types of senescent cells (Kirkland & Tchkonia, 2017). For example, unlike navitoclax (ABT263), fisetin, A1331852, A1155463 (Zhu et al., 2017,2016,2015), or Q on its own, D selectively targets senescent adipose progenitors (Zhu et al., 2015), a key cell type for adipose tissue and metabolic function (Tchkonia et al., 2013)

 

And confirmed with a quote from an older study:

http://europepmc.org...cles/pmc5391241

 

Here we demonstrate that fisetin is indeed senolytic in senescent HUVECs, but not in senescent IMR-90 cells or human preadipocytes. Interestingly, the fisetin concentrations achieved in a mouse study without causing toxicity (2.7- 349.4 μM) [30] are similar to and even higher than those we found to be senolytic in cultured HUVECs.

 

Keeping things on the bright side here.  There are new studies out by the week showing senolytics working for various disease states, and presumably aging itself.  Fisetin is still a great broad spectrum senolytic with many good targets, but D+Q also still holds the reigns for some cell types.  Apparently for metabolic dysfunction D+Q may be the better option.

Edited by OP2040, 25 March 2019 - 06:35 PM.