459 replies to this topic

[Logic]

Reason:

More Details on Bcl-2 Inhibitors as Senolytic Drugs:

http://www.longecity...enolytic-drugs/

[BieraK]

Hello guys, happy new year.
I'm experimenting with methylene blue since the last 4 months, trying to find the right dosage and learning more about the drug, there interesting things like Nrf2 enhancer, Sirt1 activation, AMPK activation, increased oxygen consumption by cells and so on.

Today I read this study (attatched at the end of the post)

 

Alternative mitochondrial electron transfer for the treatment of neurodegenerative diseases and cancers: Methylene blue connects the dots Abstract  

Brain has exceptional high requirement for energy metabolism with glucose as the exclusive energy source. Decrease of brain energy metabolism and glucose uptake has been found in patients of Alzheimer's, Parkinson's and other neurodegenerative diseases, providing a clear link between neurodegenerative disorders and energy metabolism. On the other hand, cancers, including glioblastoma, have increased glucose uptake and rely on aerobic glycolysis for energy metabolism. The switch of high efficient oxidative phosphorylation to low efficient aerobic glycolysis pathway (Warburg effect) provides macromolecule for biosynthesis and proliferation. Current research indicates that methylene blue, a century old drug, can receive electron from NADH in the presence of complex I and donates it to cytochrome c, providing an alternative electron transfer pathway. Methylene blue increases oxygen consumption, decrease glycolysis, and increases glucose uptake in vitro. Methylene blue enhances glucose uptake and regional cerebral blood flow in rats upon acute treatment. In addition, methylene blue provides protective effect in neuron and astrocyte against various insults in vitro and in rodent models of Alzheimer's, Parkinson's, and Huntington's disease. In glioblastoma cells, methylene blue reverses Warburg effect by enhancing mitochondrial oxidative phosphorylation, arrests glioma cell cycle at s-phase, and inhibits glioma cell proliferation. Accordingly, methylene blue activates AMP-activated protein kinase, inhibits downstream acetyl-coA carboxylase and cyclin-dependent kinases. In summary, there is accumulating evidence providing a proof of concept that enhancement of mitochondrial oxidative phosphorylation via alternative mitochondrial electron transfer may offer protective action against neurodegenerative diseases and inhibit cancers proliferation.

Later I found this other study (free access):
http://www.ncbi.nlm....les/PMC4017650/

Mitochondrial complex I inhibitors and forced oxidative phosphorylation synergize in inducing cancer cell death. Abstract  

Cancer cells generally rely mostly on glycolysis rather than oxidative phosphorylation (OXPHOS) for ATP production. In fact, they are particularly sensitive to glycolysis inhibition and glucose depletion. On the other hand mitochondrial dysfunctions, involved in the onset of the Warburg effect, are sometimes also associated with the resistance to apoptosis that characterizes cancer cells. Therefore, combined treatments targeting both glycolysis and mitochondria function, exploiting peculiar tumor features, might be lethal for cancer cells. In this study, we show that glucose deprivation and mitochondrial Complex I inhibitors synergize in inducing cancer cell death. In particular, our results reveal that low doses of Complex I inhibitors, ineffective on immortalized cells and in high glucose growth, become specifically cytotoxic on cancer cells deprived of glucose. Importantly, the cytotoxic effect of the inhibitors on cancer cells is strongly enhanced by forskolin, a PKA pathway activator, that we have previously shown to stimulate OXPHOS. Taken together, we demonstrate that induction in cancer cells of a switch from a glycolytic to a more respirative metabolism, obtained by glucose depletion or mitochondrial activity stimulation, strongly increases their sensitivity to low doses of mitochondrial Complex I inhibitors. Our findings might be a valuable approach to eradicate cancer cells.

Basically Methylene Blue acts as metabolic modulator that can interacts with the warburg effect on CA cells.
What can be of interest is to combine Complex I inhibitors like Metformin, then use it in conjunction with forskolin like the last study, but that can be highly enhanced with the use of Methylene Blue and Oxygen Saturation with 98% oxygen supply from bottled oxygen or from a oxygen concentrator, finally cheap drugs like mebendazole would further increase the effect.

Mebendazole is safe for healthy cells but for cancer cells it shares some of the effects with Dasatinib: http://www.longecity...e-8#entry742771

So metformin+forskolin+methylene blue+mebendazole+98% oxygen supply with an oxygen concentrator or a bottled oxygen could be another approach for killing cancer cells or senescent cells.

Sorry for the bad english and for the "simplistic" argumentation and explanation, but I'm a little hurry because of the New Year celebration.
 

Attached Files

•  yang2015.pdf   840.4KB   10 downloads

Edited by BieraK, 01 January 2016 - 12:51 AM.

[thedarkbobo]

Yes, but as it is put here http://www.longecity...methylene-blue/ MB can be also unhealthy in the long run. I plan to use MB for a few days every few weeks.

 

Regarding other posts, it's great that we see some data about Quercetin dosage (aka 500-800 mg per day of quercetin for three months to purge accumulated senile cells and then stay with a maintenance dose of 150 mg each day thereafter)

 

 

Anyone tried Navitoclax yet? I'm always baffled by how little this community reacts to such promising news as these while a thread about reservatrol can get a dozen posts per day.

 

We would need some more, safe agents for other cells and if possible custom synthesis for a group buy, since prices can be really too high for big rats.

 

[albedo]

Darryl has posted here some information on quercetin bioavailability which might also be useful here. I did not read the paper though and report only the abstract:

 

Quercetin bioavailability is associated with inadequate plasma vitamin C status and greater plasma endotoxin in adults

http://www.nutrition...0275-5/abstract

 

"Objective. Quercetin bioavailability exhibits high interindividual variation for reasons that remain unclear. We conducted a 24-h pharmacokinetic study to investigate whether individual differences in circulating antioxidants, oxidative stress and inflammation, and intestinal permeability affect quercetin bioavailability.

 

Methods. Healthy adults (n = 9 M/7 F; 34.3 ± 4.5 y; 27.0 ± 1.7 kg/m²) ingested 1095 mg quercetin aglycone with a standardized meal. Plasma antioxidants, biomarkers of oxidative stress and inflammation, and endotoxin were measured at baseline (0 h), and quercetin and its methylated metabolites isorhamnetin and tamarixetin were measured at timed intervals for 24 h.

 

Results. Plasma pharmacokinetics of quercetin, isorhamnetin and tamarixetin were highly variable between participants (CV_inter = 37–96%). Plasma vitamin C concentrations (34.6 ± 2.5 μmol/L), but no other antioxidants, were inversely correlated to the C_max and AUC_{0 to 24 h} of total quercetin (Q_total; sum of quercetin, isorhamnetin and tamarixetin; r = −0.52 to −0.53; P < 0.05). Plasma endotoxin (0.13 ± 0.01 EU/mL), a surrogate marker of intestinal permeability, was correlated to Q_total C_max (r = 0.45; P < 0.05) and tended to be correlated to Q_total AUC_{0 to 24 h} (r = 0.38; P = 0.07). Additionally, vitamin C was inversely related to C-reactive protein, myeloperoxidase, and endotoxin (r = −0.46 to −0.55; P < 0.05), whereas endotoxin was positively correlated to C-reactive protein (r = 0.73; P < 0.05).

 

Conclusions. These findings suggest that vitamin C status and plasma endotoxin may be associated with interindividual variations in quercetin bioavailability. Greater quercetin absorption and bioavailability may be associated with poor vitamin C status and increased intestinal permeability in healthy adults."

 

[Fafner55]

Experiment #8  

Building on Blogger ‘A Better World’ post #214, I included Longvida curcumin and a larger dose of pterostilbene than my previous self-experiments.  I also followed up with an additional dose of 6400 mg quercetin dihydrate 3 hours after the initial dose.  My daily log follows.

 

2016-01-09, afternoon   Starting at 3:30 in the afternoon on an empty stomach I ingested

120 mg Dasatinib

6400 mg Quercetin dihydrate

350 mg Liposomal quercetin

600 mg Sodium butyrate every 15 minutes, 8 total, starting 30 minutes before taking anything else

500 mg Pterostilbene

2000 mg Longvida curcumin

800 mg Honokiol (Magnolia bark extract)

400 mg Grape seed extract

1000 mg Green tea extract

After 3 hours, on an empty stomach I ingested another

            6400 mg Quercetin dihydrate

2016-01-09, bedtime   There has been no stomach upset, but I do feel something like a low fever.  

2016-01-10   After a night of restless sleep I awoke with mild sense of weakness and the same feelings of a low fever.

2016-01-10, bedtime   A few small, thin, dry, areas of skin on my arms, legs and face flaked off today.  This might be coincidental, but since I rarely see skin flaking, it seems likely that the treatment contributed to it.

2016-01-11   Today I feel completely normal.  Other patches of dry skin on my hands and arms flaked off this morning.  Significantly, I noticed that some pads of non-normal thick skin that appeared on my right arm a few months ago are flaking off to the extent that they are nearly disappeared.  Of all of my experiments in clearing senescent (and possibly precancerous) cells, this one is the only one that has shown obvious results.

2016-01-11, bedtime   More patches of skin flaked off, mainly on my forearms and face.  The pads of skin on my right arm are about 70% reduced in size and thickness, but they are not entirely gone.

2016-01-12, bedtime   Nearly all skin flaking is stopped.

 

It is noteworthy that Pterostilbene and quercetin have been shown to be synergistic in changing in the expression of prodeath and/or antideath Bcl-2 genes. “Association between Pterostilbene and Quercetin Inhibits Metastatic Activity of B16 Melanoma”

...because Bcl-2 overexpression can increase metastatic cell resistance against oxidative and nitrosative stress [35], changes in the expression of prodeath and/or antideath Bcl-2 genes could be related to the increase in endothelium-induced B16M-F10 cytotoxicity elicited by QUER. We studied this question by analyzing the effect of t-PTER and/or QUER on the expression of death-related Bcl-2 genes. As shown in Table 5, t-PTER increases the expression of prodeath BAX (∼ 2.2-fold) and decreases expression of antideath Bcl-2 (∼ 2.2-fold). Besides, QUER increases the expression of all pro-death genes analyzed (BAX, BAK, BAD, and BID) (e.g., BAX; ∼ 2.5-fold) and decreases the expression of all antideath genes analyzed (Bcl-2, Bcl-w, and Bcl-xL) (e.g. Bcl-2; ∼ 7.3 fold).

[Logic]

Thx for the report and info in Pterostilbene and quercetin  being synergistic Fafner55!

You may have posted the info before, but may I ask your age plz?

 

Curcumin blocks NF-kB which is upregulated during infection and plays a role in macro and/or autophagy IIRC? why are people adding it to their stacks?

Edited by Logic, 13 January 2016 - 03:14 PM.

[Fafner55]

Thx for the report and info in Pterostilbene and quercetin  being synergistic Fafner55!

You may have posted the info before, but may I ask your age plz?

 

I am 60 years old.

[BieraK]

When I took large amounts of Quercetin+Vitaminc C+Resveratrol+Pine Bark Extract+Curcumin+PQQ (all enhanced by piperine) I felt some mild fever symptoms. That was in the firt 2 or 3 ocassions however after that I never felt that anymore.

[gregcc]

Just bought some quercetin, arrives tomorrow will trial a megadose for several days as well. Would love to try dasatinib as well unfortunately out of my budget. I started a blog with a new routine i will be trying. This is possibly the only forum in the world where people might actually be interested so ill post it up here.

Edited by gregcc, 17 January 2016 - 06:51 AM.

[Logic]

Just bought some quercetin, arrives tomorrow will trial a megadose for several days as well. Would love to try dasatinib as well unfortunately out of my budget. I started a blog with a new routine i will be trying. This is possibly the only forum in the world where people might actually be interested so ill post it up here.

Check out TLR.  They have a D&Q mix for way lees than the std price.

[xEva]

 

2016-01-10, bedtime   A few small, thin, dry, areas of skin on my arms, legs and face flaked off today.  This might be coincidental, but since I rarely see skin flaking, it seems likely that the treatment contributed to it.

2016-01-11   Today I feel completely normal.  Other patches of dry skin on my hands and arms flaked off this morning.  Significantly, I noticed that some pads of non-normal thick skin that appeared on my right arm a few months ago are flaking off to the extent that they are nearly disappeared.  Of all of my experiments in clearing senescent (and possibly precancerous) cells, this one is the only one that has shown obvious results.

2016-01-11, bedtime   More patches of skin flaked off, mainly on my forearms and face.  The pads of skin on my right arm are about 70% reduced in size and thickness, but they are not entirely gone.

2016-01-12, bedtime   Nearly all skin flaking is stopped.

 

 

 

Thanks for your report.

 

Almost 2 weeks have past. Any updates?

[Fafner55]

 

 

2016-01-10, bedtime   A few small, thin, dry, areas of skin on my arms, legs and face flaked off today.  This might be coincidental, but since I rarely see skin flaking, it seems likely that the treatment contributed to it.

2016-01-11   Today I feel completely normal.  Other patches of dry skin on my hands and arms flaked off this morning.  Significantly, I noticed that some pads of non-normal thick skin that appeared on my right arm a few months ago are flaking off to the extent that they are nearly disappeared.  Of all of my experiments in clearing senescent (and possibly precancerous) cells, this one is the only one that has shown obvious results.

2016-01-11, bedtime   More patches of skin flaked off, mainly on my forearms and face.  The pads of skin on my right arm are about 70% reduced in size and thickness, but they are not entirely gone.

2016-01-12, bedtime   Nearly all skin flaking is stopped.

 

 

 

Thanks for your report.

 

Almost 2 weeks have past. Any updates?

 

 

 

2016-01-13 & 14  The skin on my chest, shoulders, upper arms and upper back is quite rough and bumpy.

2016-01-15   The rough skin on my chest, shoulders, upper arms and upper back mostly scrubbed off.  Overall, my skin appears clearer.

 

 

Experiment #9  

Cut out many of the ingredients in Experiment #8 to see if the same effect could be achieved with less and without dasatinib, of which I have only a small supply.

2016-01-17, afternoon   Starting at 3:30 in the afternoon on an empty stomach I ingested

           6400 mg Quercetin dihydrate

500 mg Pterostilbene

800 mg Honokiol (Magnolia bark extract)

(750 mg nicotinamide riboside + 16 mg C60 since this is Sunday)

After 3 hours, on an empty stomach I ingested another

           6400 mg Quercetin dihydrate

2016-01-17, bedtime   No symptoms

2016-01-18, bedtime   There is some skin flaking on my arms.

2016-01-21   Overall, the skin on my face, shoulders, chest, arms and legs looks clear and younger.

 

 

 

 

Edited by Fafner55, 25 January 2016 - 10:23 PM.

[Fafner55]

I can further add that my skin all over my body looks better than it has in many years.  What has changed is that much sun damaged skin is now replaced by more uniform, smooth skin.  What has not changed are age-spots, fine wrinkles on my hands and some crepiness in the skin of inside of my arms.

 

There has been enough improvement to my skin that I don't feel that I will be able to see much difference in future tests.  Hopefully others over the age of 50 or 60 will do similar test and confirm my results.  However, I am concerned that this treatment could be dangerous to people with a high percentage of senescent cells, such as those over the age of 75 or 80, or those who might have been longtime smokers and have COPD.   Please be careful.

[Logic]

A study on Q absorption that may be helpful:

 

We have tested the absorptive efficiency of...quercetin...10 mg/70 kg...

randomly administered at 4-week intervals in three different matrices: white wine (11.5% ethanol), grape juice, and vegetable juice/homogenate. Blood was collected at zero time and at four intervals over the first four hours after consumption; urine was collected at zero time and for the following 24-h.

The sums of free and conjugated polyphenols were measured in blood serum and urine by a gas-chromatographic method...[Quercetin] was present in serum and urine predominantly as glucuronide and sulfate conjugates, reaching peak concentrations in the former around 30-min after consumption...

The free polyphenols accounted for... 7.2 to 26.9% (quercetin) of the peak serum concentrations...

(urine 24-h excretion 2.9%-7.0% of dose consumed)...

http://www.sciencedi...009912002003971

 

[Fafner55]

To understand the importance of clearing senescent cells it is useful to compare the percentage of senescent cells in a human body to survival curves.

 

After childbearing age, about 35 years, the mortality rate in the US is accurately modeled by the simple Gompertz equation given by

https://en.wikipedia.org/wiki/Baboon

 

m(t) = A exp(G t)

 

where

m(t) = the mortality rate as a function of age (t) in years

A = 8.84 is the extrapolated constant to birth or maturity for a population of 100,000.

G = 0.0800 is the exponential (Gompertz) mortality rate coefficient determined from regression analysis.

 

The black line in the figure below represents the Gompertz function extrapolated from the mortality rates after maturity.

 

 

<Gompertz mortality curve.jpg> 

 Gompertz mortality curve.jpg   16.72KB   6 downloads

 

Mortality rates, expressed in deaths per 100,000 people, as a function of age for the 2002 US population.

 

 

The Gompertz equation for modeling the death rate can be integrated and rearranged to derive the probability of survival.   The survival model of living to age “t” is derived from the standard Gompertz equation as

 

s(t) = T exp(B (1 - exp(G t)))

 

where

s(t) = the survival rate as a function of age t in years

T = the initial population = 100 when calculating the percentage

B = (A/100,000 ) /  G

and A and G are defined above

 

In the case of dermal fibroblasts in vivo for baboons, the progression of morphological changes and transition from mitotic to senescence is measured as shown in the table below. “Cellular Senescence in Aging Primates” (Herbig et al, 2006) http://www.biodados.icb.ufmg.br/cromatina/dna07/telomere.pdf

 

<Senescence in baboon populations.jpg> 

 Senescence in baboon populations.jpg   31.41KB   6 downloads

 

Baboons in captivity have been known to live up to 45 years.  https://en.wikipedia.org/wiki/Baboon

Assuming a 45 year old baboon is equivalent to a 100 year old human and applying exponential regression to the above data, an equation for the rate of senescence in dermal fibroblasts is determined to be

 

s(t) = 1.6455exp(0.0465t)

 

m(t) = 100 - s(t)

 

where

s(t) = the percentage rate of cellular senescence (i.e., post-mitotic) in years, t

m(t) = the percentage rate of remaining mitotic cells

 

It should be noted that the Herbig data for cellular senescence was taken from a small sample of baboons with significant range of values at older ages.

 

In the case of human dermal fibroblasts in vivo, the progression of morphological changes and transition from mitotic to senescence is measured as shown in the table below.  “Fibroblasts in normal and pathological terminal differentiation, aging, apoptosis and transformation” (Bayreuther et al, 1992) http://www.ncbi.nlm.nih.gov/pubmed/18647676

 

<Fibroblasts samples from people of different ages.jpg>  

 Fibroblasts samples from people of different ages.jpg   34.01KB   6 downloads

 

Fibroblasts samples from people of different ages.  Cell types I-III are mitotic and V-VI are post-mitotic.  

 

Applying exponential regression to the above data for post-childbearing ages 50 and older, an equation for the rate of senescence in dermal fibroblasts is determined to be

 

s(t) = 9.4474exp(0.0247t)

 

m(t) = 100 - s(t)

 

where

s(t) = the percentage rate of cellular senescence (i.e., post-mitotic) in years, t

m(t) = the percentage rate of remaining mitotic cells

 

When the remaining percentage of mitotic cells is plotted against human survival (the % of a population remaining) the correlation is compelling.

 

<Senescent cells vs survival.jpg> 

 Senescent cells vs survival.jpg   37.7KB   6 downloads

 

It is likely that extrapolating the Herbig and Bayreuther estimates from dermal fibroblasts to the entire body overstates the percentage of total percentage of senescent cells,  but these results do explain the poor condition of skin in the extreme elderly.

 

The high correlation between cellular senescence and mortality implies that cellular senescence is a proximate root cause of many causes of mortality.  One can also infer that if a body was cleared of senescent cells, the normal regenerative process would replace those cells with healthy ones and organismal aging would to some extent be reversed.

 

 

 

 

[PWAIN]

We need to identify a few more of these.

[aribadabar]

 

Baboons in captivity have been known to live up to 45 years.  https://en.wikipedia.org/wiki/Baboon

Assuming a 45 year old baboon is equivalent to a 100 year old human and applying exponential regression to the above data, an equation for the rate of senescence in dermal fibroblasts is determined to be

 

s(t) = 1.6455exp(0.0465t)

 

 

Thanks for the informative ideas.

 

I have several questions with the above assumption:

1) Would the senescent progression be the same over 100-year period? I.e. shouldn't the slope be less steep as it is over a longer timeframe (compared to baboon lifespan) to reach same level of decrepitude and burden in function or the rates of progression are already normalized and the longer timeframe is already taken into account?

2) If 1) is already addressed in that formula, isn't it more accurate to equate the 45 baboon years to 115-122 as that is the max lifespan of humans?

Edited by aribadabar, 27 January 2016 - 05:43 PM.

[Fafner55]

 

 

Baboons in captivity have been known to live up to 45 years.  https://en.wikipedia.org/wiki/Baboon

Assuming a 45 year old baboon is equivalent to a 100 year old human and applying exponential regression to the above data, an equation for the rate of senescence in dermal fibroblasts is determined to be

 

s(t) = 1.6455exp(0.0465t)

 

 

Thanks for the informative ideas.

 

I have several questions with the above assumption:

1) Would the senescent progression be the same over 100-year period? I.e. shouldn't the slope be less steep as it is over a longer timeframe (compared to baboon lifespan) to reach same level of decrepitude and burden in function or the rates of progression are already normalized and the longer timeframe is already taken into account?

2) If 1) is already addressed in that formula, isn't it more accurate to equate the 45 baboon years to 115-122 as that is the max lifespan of humans?

 

 

Regarding your first question, one must be careful not to over interpret these curves.

1. There is a large variation in the burden of senescent cells in 25-30 year baboons.  If a larger sample could be taken the auto regression curve would likely be different.
2. These curves will not hold up if they are extrapolated very far out of their measurement ranges.  For instance, the mitotic cell curves for the Herbig and Bayreuther data would be a nonsensical negative number before the age of 100 if it were extended.

3. Herbig and Bayreuther could be measuring slightly different things: Herbig looked at a 53BP1 marker, while Bayreuther measured something else (needs confirmation).

 

 

As for question 2, it is difficult to compare a small population of baboons to a large population of people.

 

From https://en.wikipedia...upercentenarian the upper end estimate of supercentarians in the world is 450 people (those living past 110 years out of 7.4 billion).  That is 6 per 10^^9 people, certainly very rare.  In comparison, the number of baboons in captivity is much smaller.  The odds of the longest lived baboon in captivity being more similar to a 100 year old person is much more likely than that lucky baboon being the equivalent of a 115 to 122 year old human.

 

That said, the question of how to map baboon lifespans to humans is important.  Should it be based on the age of sexual maturity, average lifespan, a LMS fit of mortality curves, maximum lifespan or something else?

 

I choose maximum lifespan and an age of 100 because

1. The comparison is made to the Gompertz equation, which holds after the childbearing age of 35 or so.
2. By the Grompertz equation, nearly all humans are dead by the age of 100.
3. The maximum lifespan data was available and reliable
4. The population of baboons in captivity is small.

Certainly other assumptions can be made and these equations for the rates of cellular senescence will shift.  Even so, the conclusions will remain the same: the correlation between survival rates and remaining percentages of mitotic cells will be high, and cellular senescence proximate cause of organismal aging and underlies many causes of mortality.  Cellular senescence is not the end-all explanation for aging, but these curves should highlight to the important role they play and the imperative for clearing them.

[Fafner55]

Apologies to all.  My last post had a math error and should have stated that the incidence of supercentarians is about 60 per 10^^9 people, which is still rare.

Fafner55

 

[Greg Cornes]

Interesting read for anyone interested in quercetins effects on cognition. 

 

http://biomedjournal...tory-animals-2/

 

Similar dosages were administered in this study.

 

However also found this study which suggests the opposite. 

 

http://www.ncbi.nlm....ubmed/20466049/

[Fafner55]

Experiment #10  

Another attempt to see if a senolytic benefit to my skin could be achieved without dasatinib.  

2016-01-31, morning   After coffee and an English muffin I ingested

800 mg Honokiol (Magnolia bark extract)

400 mg Grape seed extract

1000 mg Quercetin Phytosome (By Thorne Research.  Quercetin Phytosome is quercetin bound to phosphatidylcholine for better absorption.)

300 mg Pterostilbene

750 mg Nicotinamide riboside + 16 mg C60 since this is Sunday

2016-01-31, mid-afternoon  No adverse symptoms.  The thick patches of skin on my right arm that shrank about 70% in Experiment #8 are noticeably reduced further in the 5.5 hours since I started this morning, maybe by 30-40%.   I did not see this effect with Experiment #9.  I can’t be sure that this benefit is in fact senolytic or if it is just strongly apoptotic to precancerous cells.  I don't notice any skin flaking. In any case, it is clearly beneficial to my skin.

[niner]

2016-01-31, mid-afternoon  No adverse symptoms.  The thick patches of skin on my right arm that shrank about 70% in Experiment #8 are noticeably reduced further in the 5.5 hours since I started this morning, maybe by 30-40%.   I did not see this effect with Experiment #9.  I can’t be sure that this benefit is in fact senolytic or if it is just strongly apoptotic to precancerous cells.  I don't notice any skin flaking. In any case, it is clearly beneficial to my skin.

 

5.5 hours seems too fast for that amount of shrinking of a skin lesion, particularly without flaking or peeling.  Are you taking photographs?

[Fafner55]

 

2016-01-31, mid-afternoon  No adverse symptoms.  The thick patches of skin on my right arm that shrank about 70% in Experiment #8 are noticeably reduced further in the 5.5 hours since I started this morning, maybe by 30-40%.   I did not see this effect with Experiment #9.  I can’t be sure that this benefit is in fact senolytic or if it is just strongly apoptotic to precancerous cells.  I don't notice any skin flaking. In any case, it is clearly beneficial to my skin.

 

5.5 hours seems too fast for that amount of shrinking of a skin lesion, particularly without flaking or peeling.  Are you taking photographs?

 

 

5.5 hours is surprising fast.  I was hesitant to post this result because it didn't seem credible, but that's what happened.  It is possible that the patch of affected skin was particularly vulnerable to the apoptotic pathways put in play by these supplements.  Last night, after 12 hours, those patches were further reduced in size and thickness to about 50% of their starting size.  This morning (2015-02-01) they appear the same as last night.

 

It will be interesting to see if others get results from the same treatment.

 

Periodically I take pictures of my left hand to measure any benefit to wrinkles.  It is quite difficult to get consistent lighting and comparable photos.  

[Fafner55]

In case anyone missed this,

 

“Naturally occurring p16Ink4a-positive cells shorten healthy lifespan” (2016)

Cellular senescence, a stress-induced irreversible growth arrest often characterized by expression of p16Ink4a (encoded by the Ink4a/Arf locus, also known as Cdkn2a) and a distinctive secretory phenotype, prevents the proliferation of preneoplastic cells and has beneficial roles in tissue remodelling during embryogenesis and wound healing. Senescent cells accumulate in various tissues and organs over time, and have been speculated to have a role in ageing. To explore the physiological relevance and consequences of naturally occurring senescent cells, here we use a previously established transgene, INK-ATTAC, to induce apoptosis in p16Ink4a-expressing cells of wild-type mice by injection of AP20187 twice a week starting at one year of age. We show that compared to vehicle alone, AP20187 treatment extended median lifespan in both male and female mice of two distinct genetic backgrounds. The clearance of p16Ink4a-positive cells delayed tumorigenesis and attenuated age-related deterioration of several organs without apparent side effects, including kidney, heart and fat, where clearance preserved the functionality of glomeruli, cardio-protective KATP channels and adipocytes, respectively. Thus, p16Ink4a-positive cells that accumulate during adulthood negatively influence lifespan and promote age-dependent changes in several organs, and their therapeutic removal may be an attractive approach to extend healthy lifespan.

 

Discussion

Here we eliminated p16Ink4a+ cells from non-progeroid mice using ATTAC to begin to address how senescent cells influence health and lifespan. We observe median lifespan extensions in AP-treated ATTAC mice on two distinct genetic backgrounds and diets, indicating that age-related accumulation of p16Ink4a+ cells negatively affects longevity. It will be useful to optimize senescent cell removal protocols and methods further because the longevity of male C57BL/6 mice seemed negatively affected by repetitive vehicle injection stress, and because clearance was partial and several key tissues were refractory to clearance, including liver and colon. This would be particularly relevant for further studies of maximum lifespan, extension of which is thought to be reflective of a more generalizable ‘anti-ageing’ effect.

 

These methodological caveats aside, we find that removal of p16Ink4a+ cells from midlife on delays the progression of neoplastic disease irrespective of genetic background and diet. Clearance had no effect on the spectrum or incidence of post-mortem tumours, which is consistent with the hypothesis that senescent cells help create a tissue microenvironment that promotes tumour progression1. In addition, several age-dependent changes, including alterations in fat, eye, heart and kidney, as well as the age-dependent decline in spontaneous activity and exploratory behaviour, occur more slowly during a six-month period of p16Ink4a+ cell clearance starting at midlife. Complementary mechanistic studies suggest that local p16Ink4a+ cells in fat, kidney and heart exert their effects through distinct mechanisms, involving progenitor cell dysfunction, RAAS overactivation, and Sur2a downregulation, respectively, although systemic effects cannot be excluded given the global nature of the clearance method applied here.

 

Thus, our data best fit a model in which p16Ink4a+ cells act to shorten healthy lifespan by promoting tumour progression and age-dependent changes that functionally impair certain tissues and organs, including vital organs such as kidney and heart. This, together with the key observation that elimination of p16Ink4a+ cells is not associated with any overt detrimental effects raises the possibility that this approach may be useful to treat aspects of age-related functional decline, age-related diseases that involve senescent cells, or side effects of therapies that create senescent cells.

[Bryan_S]

 

In case anyone missed this,

 

“Naturally occurring p16Ink4a-positive cells shorten healthy lifespan” (2016)

 

It also made its rounds thru some other publications and the concept is gaining weight. Here are a few links that hit all about the same time. The last link "Chromosomes reconfigure as cell division ends" tells the story of the fundamental question about what genetic changes come along with senescence. I'm pretty sure this is now getting the academic recognition it deserves.

 

Scientists boost lifespan of mice by deleting defective cells

February 4, 2016 by Richard Faragher

 

http://phys.org/news...e-deleting.html

 

https://theconversat...ive-cells-54068

 

Ageing: Out with the old

http://www.nature.co...ture16875.html 

 

 

Destroying worn-out cells makes mice live longer

Elegant experiment confirms that targeting senescent cells could treat age-related diseases.

 

Ewen Callaway

03 February 2016

http://www.nature.co...-longer-1.19287

 

Chromosomes reconfigure as cell division ends

http://phys.org/news...l-division.html

Edited by Bryan_S, 06 February 2016 - 08:10 AM.

[Fafner55]

Summarizing “Naturally occurring p16Ink4a-positive cells shorten healthy lifespan”,

 

Improved

• Lifespan (17-35% medium improvement depending on the mouse strain and diet.  There was also an improvement in maximum lifespan.)

• Heart (no significant differences in heart rate, left ventricular mass, thickness and diameter, cardiac stress resistance, ejection fraction and fractional shortening in treated 18-month old treated mice compared to 12-month-old mice)

• Kidneys  (treatment markedly reduced glomerulosclerosis independent of sex and genetic background, which correlated with attenuated age-related increases in blood urea nitrogen, indicating preserved kidney function)

• Fat (substantially restored subcutaneous and visceral fat in 18 month old mice to that of 12 month olds)

• Inflammation (reduced markers in gastrocnemius, eye, kidney, heart (atria), spleen, lung, liver, skeletal muscle  and colon tissues)

• Cancer (delayed)

• Cataracts (delayed)

 

Did Not Improve

• Coordination

• Memory

• Exercise ability

• Haematological parameters and age-related changes in leukocyte populations (White blood cell count, platelet count, red blood cell count, haemoglobin concentration, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, neutrophils, lymphocytes, basophils, monocytes and eosinophils)

• Somatotrophic axis signalling (glucose levels following intraperitoneal glucose administration after an overnight fast)

• Cardiac morphology and function in ‘resting’ mice

• Wound healing and tissue fibrosis

[stefan_001]

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2016-01-31, mid-afternoon No adverse symptoms. The thick patches of skin on my right arm that shrank about 70% in Experiment #8 are noticeably reduced further in the 5.5 hours since I started this morning, maybe by 30-40%. I did not see this effect with Experiment #9. I can’t be sure that this benefit is in fact senolytic or if it is just strongly apoptotic to precancerous cells. I don't notice any skin flaking. In any case, it is clearly beneficial to my skin.

5.5 hours seems too fast for that amount of shrinking of a skin lesion, particularly without flaking or peeling. Are you taking photographs?

5.5 hours is surprising fast. I was hesitant to post this result because it didn't seem credible, but that's what happened. It is possible that the patch of affected skin was particularly vulnerable to the apoptotic pathways put in play by these supplements. Last night, after 12 hours, those patches were further reduced in size and thickness to about 50% of their starting size. This morning (2015-02-01) they appear the same as last night.

It will be interesting to see if others get results from the same treatment.

Periodically I take pictures of my left hand to measure any benefit to wrinkles. It is quite difficult to get consistent lighting and comparable photos.

Honokiol has caused shrinking of some patches in my experience. When using it as topical some patches/flakes peel off and flatten . Interestingly some don't. I attributed that to apoptotic pathway.

What I find curious is that you have not seen improvement in wrinkles. Makes me wonder what is really happening perhaps it's all and only apoptotic pathway action on pre cancerous cells either benign or malicious.

Edited by stefan_001, 06 February 2016 - 06:37 PM.

[niner]

What I find curious is that you have not seen improvement in wrinkles. Makes me wonder what is really happening perhaps it's all and only apoptotic pathway action on pre cancerous cells either benign or malicious.

 

Senescent cells aren't a big contributor to wrinkles.   The main contributors are photodamage and glycation.   Fafner's results suggest that pre-cancerous lesions (such as keratoses) are rich in senescent cells.  That makes a lot of sense.

[stefan_001]

What I find curious is that you have not seen improvement in wrinkles. Makes me wonder what is really happening perhaps it's all and only apoptotic pathway action on pre cancerous cells either benign or malicious.

Senescent cells aren't a big contributor to wrinkles. The main contributors are photodamage and glycation. Fafner's results suggest that pre-cancerous lesions (such as keratoses) are rich in senescent cells. That makes a lot of sense.

I didn't think of that but yes indeed makes a lot if sense. Thanks for pointing out. It may also mean that glycation is potentially a bigger problem in the long run.

[Fafner55]

.
.

 

 

2016-01-31, mid-afternoon No adverse symptoms. The thick patches of skin on my right arm that shrank about 70% in Experiment #8 are noticeably reduced further in the 5.5 hours since I started this morning, maybe by 30-40%. I did not see this effect with Experiment #9. I can’t be sure that this benefit is in fact senolytic or if it is just strongly apoptotic to precancerous cells. I don't notice any skin flaking. In any case, it is clearly beneficial to my skin.

5.5 hours seems too fast for that amount of shrinking of a skin lesion, particularly without flaking or peeling. Are you taking photographs?

5.5 hours is surprising fast. I was hesitant to post this result because it didn't seem credible, but that's what happened. It is possible that the patch of affected skin was particularly vulnerable to the apoptotic pathways put in play by these supplements. Last night, after 12 hours, those patches were further reduced in size and thickness to about 50% of their starting size. This morning (2015-02-01) they appear the same as last night.

It will be interesting to see if others get results from the same treatment.

Periodically I take pictures of my left hand to measure any benefit to wrinkles. It is quite difficult to get consistent lighting and comparable photos.

Honokiol has caused shrinking of some patches in my experience. When using it as topical some patches/flakes peel off and flatten . Interestingly some don't. I attributed that to apoptotic pathway.

What I find curious is that you have not seen improvement in wrinkles. Makes me wonder what is really happening perhaps it's all and only apoptotic pathway action on pre cancerous cells either benign or malicious.

 

 

I agree that my experiments 9 and 10 likely invoked apoptotic pathway action on precancerous cells and showed no clear indication of senolytic action.  Experiment 8 which included dasatinib was different.  The substantial amount of sun damaged skin that was shed as a result of that experiment must have resulted from apoptosis of senescent cells in addition to pre-cancerous cells.  The skin on my chest, shoulders and back remains remarkably clear and smooth.

 

As for wrinkles, it has been 24 days since Experiment 8 (Jan 13) and I continue to see no effect on fine wrinkles.  My expectation was that there would be remodeling of skin tissues as cells turned over, collagen gene expression hopefully restored and as fat layers returned to more youthful levels.  But, I see no evidence of any of this.