134 replies to this topic

[Gerald W. Gaston]

By alternating periods of protein restriction with periods of protein abundance, repeated induction of autophagy could clear the aggregates associated with AD, PD, HD and ALS and maybe even clear cryptic infections. A protein cycling diet is much easier to follow than a calories restriction diet.

Remig,
It seems I'm been doing something similar to what you describe in my attempt to reduce dietary AGEs... I call my protein days my 'kill' days (as if I'm hunting down and killing game). These are my twice a week anabolic workout days (and the day after). This is on a 8 day cycle with 2 days of protein reduction in between each workout day, so my two workout days move each week.

I see this is your first post... welcome to ImmInst!

[niner]

Non-selective macro and micro autophagy is used by cells when the supply of nutrients by the body is inadequate. The body must supply calories, essential fatty acids, and the essential amino acids required for protein synthesis. Whenever the supply is inadequate, the cell must then acquire the missing nutrients by recycling some portion of itself.

In non-selective autophagy (not CMA), some portion of the cell's cytoplasm is enclosed in a membrane. The membrane may be the lysosome's own membrane (in which case it is termed microautophagy) or another membrane generated for just that purpose (in which case it is call macroautophagy.) In macroautophagy, the vesicle so formed then merges with lysosomes and the end result of the two forms of autophagy is the same, some portion of the cytoplasm, including even entire organelles such as mitochondria, is digested and the constituent molecules made available for the anabolic processes of the cell.

A shortage of any essential structural nutrient induces autophagy. Creating a shortage of essential amino acids (leucine in particular) is a standard way to induce to induce autophagy in free cell culture (nitrogen starvation). Anybody can do the same by restricting protein in the diet for some period of time. I have an on-line monograph on just this subject if anyone is interested.

By alternating periods of protein restriction with periods of protein abundance, repeated induction of autophagy could clear the aggregates associated with AD, PD, HD and ALS and maybe even clear cryptic infections. A protein cycling diet is much easier to follow than a calories restriction diet.

Welcome to ImmInst, remiq. I looked at your monograph, it's great! I think that a lot of the people here will find it most interesting. I strongly recommend it.

[FunkOdyssey]

That monograph is excellent, amazing work, I'm surprised you are not attempting to sell that. When do you plan to finish the sections on intracellular infection? I am very interested to see what you come up with.

[Gerald W. Gaston]

That monograph is excellent, amazing work, I'm surprised you are not attempting to sell that. ...

http://www.lulu.com/content/5429773

[FunkOdyssey]

Ahh. And I assume the missing sections are present in the for-sale book?

[Gerald W. Gaston]

Ahh. And I assume the missing sections are present in the for-sale book?

That I don't know... perhaps Ron will return... but then the ebook is only $5 so easy enough to find out.

[100YearsToGo]

Interestingly there is a human desease that is caused by a mutation in the lamp2 gene. This causes defects in protein synthesis of Lamp 2A and Lamp2B. It's called DANON desease.

Severe cardiomyopathy and variable skeletal muscle weakness are constant features and mental retardation is very frequently associated.

The median age at diagnosis was 16 years

Sounds like acelerated old age at least partly because of protein acumulation due to Lamp2A deficiency. No CMA.

Edited by 100YearsToGo, 24 February 2009 - 05:14 PM.

[remig]

Ahh. And I assume the missing sections are present in the for-sale book?

That I don't know... perhaps Ron will return... but then the ebook is only $5 so easy enough to find out.

Oops, I forgot to mark that chapter as published. That has now been fixed. The on-line version is more up-to-date than the paper version. The paper version only exists for pressing my ideas on people who can't read computer monitors. I have a blog that chronicles the updates to the book...

[100YearsToGo]

Ahh. And I assume the missing sections are present in the for-sale book?

That I don't know... perhaps Ron will return... but then the ebook is only $5 so easy enough to find out.

Oops, I forgot to mark that chapter as published. That has now been fixed. The on-line version is more up-to-date than the paper version. The paper version only exists for pressing my ideas on people who can't read computer monitors. I have a blog that chronicles the updates to the book...

I liked your sections about cell biology. Pretty clear and concise. I think Protein cycling has a good chance to activate micro an macro autophagy. But what are your ideas on CMA? Cuervo has some papers that show that if you take CMA away, macro autophagy will take over, but cells and organs will become more vulnerable to stress as compensation is incomplete.

Have you done any (unpublished) research on CMA? or do you think protein cycling would take care of this as well?

Edited by 100YearsToGo, 25 February 2009 - 12:25 AM.

[bgwithadd]

Won't this cause some organ and brain damage, though? What's the point of living slightly longer if you are less functional?

[niner]

Won't this cause some organ and brain damage, though? What's the point of living slightly longer if you are less functional?

On the contrary, it should prevent organ and brain damage. You're replacing damaged proteins with new ones; that's a good thing.

[bkaz]

Remiq, this is extremely interesting, but I don't see why exact timing is important for non-protein-restriction upregulators of autophagy (such as Curcumin)? My understanding is, autophagy dumps amino acids into cytoplasm, & cell can use them to synthesize new proteins at the same time?

[bgwithadd]

Won't this cause some organ and brain damage, though? What's the point of living slightly longer if you are less functional?

On the contrary, it should prevent organ and brain damage. You're replacing damaged proteins with new ones; that's a good thing.

Aren't these cells getting destroyed, though? That's great if they get replaced, but there's no guarantee that will happen.

[remig]

.I liked your sections about cell biology. Pretty clear and concise. I think Protein cycling has a good chance to activate micro an macro autophagy. But what are your ideas on CMA? Cuervo has some papers that show that if you take CMA away, macro autophagy will take over, but cells and organs will become more vulnerable to stress as compensation is incomplete.

Have you done any (unpublished) research on CMA? or do you think protein cycling would take care of this as well?

CMA and UPS are the constitutive recycling processes of the cell and their upregulation may be beneficial though I have no idea how to do that safely. Certainly protein cycling shouldn't take CMA away.

I know Cuervo finds that only CMA promotion extends mice life-spans. My orientation is towards preventing certain diseases conditions rather than extending life-spans. Nevertheless may I just suggest that longevity studies on mice may have interpretation problems in consequence of the antipodal evolutionary strategies of mice and men. Humans take a software approach to adaptation whereas mice take a hardware approach.

Mice are genetically very plastic. Subpopulations readily develop that are incapable of reproducing with neighboring populations because of chromosome splits and joins. Mutations and consequent cancers are very common in mice. This all supports their strategy of evolving new species to exploit new niches. This strategy also requires short life spans and high reproductive rates to achieve maximum rates of evolution. Perhaps mice even have mechanisms to guarantee short life spans.

Humans, on the other hand, use cultural means to exploit new niches. This strategy requires long life-spans and slow reproductive rates, and what applies to mice may sometimes not apply to men.

Edited by remig, 25 February 2009 - 05:13 PM.

[remig]

Aren't these cells getting destroyed, though? That's great if they get replaced, but there's no guarantee that will happen.

Cells are not getting destroyed.

If you google autophagy, much of what now comes up concerns cancer applications. One of the uses the cell makes of autophagy is apoptosis, programmed cell death. A cell is often called upon to sacrifice its self for the greater good of the body. Maybe it is harboring a virus. Or maybe it is between two developing fingers that need to separate. This autophagy apoptosis mechanism can be triggered in some cancer cell types to eliminate the cancer. In other cancer cell types, promoting autophagy actually may rescue the cancer cell from the artificial starvation condition created by chemo. Regardless, apoptosis is NOT the consequence of autophagy in normal cells in response to nutrient restriction. Only a small percentage of the trillions of molecules in a cell are sacrificed in a single autophagy event to keep the cell going. I advocate repeated cycles of autophagy and recovery to accumulate this small percentage to a value that might be significant in terms of clearing protein aggregates.

Edited by remig, 25 February 2009 - 05:36 PM.

[remig]

Remiq, this is extremely interesting, but I don't see why exact timing is important for non-protein-restriction upregulators of autophagy (such as Curcumin)? My understanding is, autophagy dumps amino acids into cytoplasm, & cell can use them to synthesize new proteins at the same time?

I think a distinction needs to be made between 'promoters' and 'upregulators'. Upregulation refers to a change in the mix of proteins synthesized by the cell in response to some change in conditions. Curcumin promotes autophagy by interfering with insulin's interaction with mTor. The cell then initiates autophagy because it thinks its calorie supply will be insufficent. The amino acids and fats so released may then be burned for their calorie content. This autophagy is its short-term response. Its long term response may be to upregulate (increase) the proteins of the mTor system to dilute curcumin's interference especially when it realizes it has been fooled. If this in fact happens, then curcumin is actually a downregulator of autophagy. Since many substances besides curcumin might be expected to also interact with mTor and fool the cell, I would expect downregulation might very well occur since energetics is so crucial to the cell and, therefore, its feedback systems should be highly robust and alert to interferences like curcumin.

If curcumin is both a promoter and downregulator of autophagy then its use needs to stopped after a promotion event to prevent the later-acting downregulation event. In other words it needs to be cycled. If other strategies to promote autophagy such as protein cycling are concurrently being employed, then the two should be synchronized.

[bkaz]

I think a distinction needs to be made between 'promoters' and 'upregulators'. Upregulation refers to a change in the mix of proteins synthesized by the cell in response to some change in conditions. Curcumin promotes autophagy by interfering with insulin's interaction with mTor. The cell then initiates autophagy because it thinks its calorie supply will be insufficent. The amino acids and fats so released may then be burned for their calorie content. This autophagy is its short-term response. Its long term response may be to upregulate (increase) the proteins of the mTor system to dilute curcumin's interference especially when it realizes it has been fooled. If this in fact happens, then curcumin is actually a downregulator of autophagy. Since many substances besides curcumin might be expected to also interact with mTor and fool the cell, I would expect downregulation might very well occur since energetics is so crucial to the cell and, therefore, its feedback systems should be highly robust and alert to interferences like curcumin.

If curcumin is both a promoter and downregulator of autophagy then its use needs to stopped after a promotion event to prevent the later-acting downregulation event. In other words it needs to be cycled. If other strategies to promote autophagy such as protein cycling are concurrently being employed, then the two should be synchronized.

Appreciate detailed explanation. It's a bit counterintuitive though, curry & other polyphenols are always eaten with food, & most need fat to be absorbed. You say that amino acids are available ~6 hrs after a meal, while curcumin & resveratrol peak ~0.5 hr after ingestion, wouldn't that be enough of a gap? Your idea about developing tollerance makes sense, but perhaps using a variety of polyphenols (as in curry) &| cycling them can counteract that? Also, the cell is not actually starving, it has glucose, why would it burn amino acids?

[100YearsToGo]

Trehalose induces autophagy in persistently prion-infected neuronal cells:

http://www.ncbi.nlm....Pubmed_RVDocSum

[100YearsToGo]

Interesting:

β-hydroxybutyrate (BOH) stimulates CMA.:

"We found that physiological concentrations of β- hydroxybutyrate (BOH) induced proteolysis in cells maintained in media with serum and without serum; however, acetoacetate only induced proteolysis in cells maintained in media with serum."

http://www.jbc.org/c...502456200v1.pdf

β-hydroxybutyrate and acetoacetate (ketones) circulate in your body usually under conditions of prolonged starvation. It is also used to make biodegradable plastic.

Coincidentally ketone bodies also protect neurons (a la trehalose) in models of alzheimer and parkinson. Is it because of autophagy?
http://www.pnas.org/...97/10/5440.full

Edited by 100YearsToGo, 27 February 2009 - 12:43 AM.

[FunkOdyssey]

Interesting:

β-hydroxybutyrate (BOH) stimulates CMA.:

"We found that physiological concentrations of β- hydroxybutyrate (BOH) induced proteolysis in cells maintained in media with serum and without serum; however, acetoacetate only induced proteolysis in cells maintained in media with serum."

http://www.jbc.org/c...502456200v1.pdf

β-hydroxybutyrate and acetoacetate (ketones) circulate in your body usually under conditions of prolonged starvation. It is also used to make biodegradable plastic.

Coincidentally ketone bodies also protect neurons (a la trehalose) in models of alzheimer and parkinson. Is it because of autophagy?
http://www.pnas.org/...97/10/5440.full

All roads lead to low carb (ketogenically low in this case)?

[geddarkstorm]

All roads lead to low carb (ketogenically low in this case)?

Or low carbs to increase ketone production? Hmm... A cursory search hasn't turned up anything for me, but can beta-hydroxybutyrate simply be ingested to the same effect, or does it have to be produced in a low glucose environment? I wonder.

Ketogenic diets also seem to be very good for the brain when it comes to increasing energy availability. I am highly intrigued, especially if mixed with trehalose. Add in resveratrol and/or carnitine to counter any adverse effects from the high fat intake in the ketogenic diet, and we may be on to an interesting regiment system... If ketone intake itself can simulate the same, that would be even better I think.

Edit: Apparently one can take beta-hydroxybutyrate orally and increase the circulating serum levels, and in the brain: "In cerebrospinal fluid, concentrations of beta-OHB increased to levels comparable to a 24- to 40-h fast, after single dosages of 4 and 8 g, respectively."

This is a pretty high dosage though, but if beta-HB is the main effector and doesn't need acetoacetate too, then it might be worth checking out.

Edited by geddarkstorm, 27 February 2009 - 01:38 AM.

[100YearsToGo]

All roads lead to low carb (ketogenically low in this case)?

Or low carbs to increase ketone production? Hmm... A cursory search hasn't turned up anything for me, but can beta-hydroxybutyrate simply be ingested to the same effect, or does it have to be produced in a low glucose environment? I wonder.

Ketogenic diets also seem to be very good for the brain when it comes to increasing energy availability. I am highly intrigued, especially if mixed with trehalose. Add in resveratrol and/or carnitine to counter any adverse effects from the high fat intake in the ketogenic diet, and we may be on to an interesting regiment system... If ketone intake itself can simulate the same, that would be even better I think.

Edit: Apparently one can take beta-hydroxybutyrate orally and increase the circulating serum levels, and in the brain: "In cerebrospinal fluid, concentrations of beta-OHB increased to levels comparable to a 24- to 40-h fast, after single dosages of 4 and 8 g, respectively."

This is a pretty high dosage though, but if beta-HB is the main effector and doesn't need acetoacetate too, then it might be worth checking out.

It seems safe for at least a couple of months use...that would be enough to clear up all those pesky misfolded stuff in your cells and also give your mitochondria a big boost.

"During the 5- to 7-month treatment period, octreotide demand and the frequency and degree of hypoglycemic episodes did not change. This strongly suggests that additional insulin secretion was not induced as a possible side effect of DL sodium
-hydroxybutyrate. Cardiocirculatory changes did not occur and preexisting ultraonographic signs of hypertrophic cardiomyopathy even improved during the time course of observation in both patients. Cardiotoxicity as possible side effect of DL sodium
-hydroxybutyrate is thus unlikely. Vomiting or loss of "orality" was preexisting to the administration of DL sodium
-hydroxybutyrate and did not deteriorate upon treatment."

http://www.pedresearch.org/pt/re/pedresear...#33;8091!-1

Edited by 100YearsToGo, 27 February 2009 - 03:09 PM.

[100YearsToGo]

Cuervo receives more $$$ for research on autophagy:

http://www.genengnew...x?name=50251597

The researchers wil:

look at the role of two different types of autophagy in liver and brain function as well as immunity, under normal and stressful conditions <LI>analyze how these two types of autophagy change as the liver, brain, and immune system age <LI>determine how changes in autophagy that occur with age contribute to the aging of the entire organism, to the gradual deterioration of cognitive function, to the failure with age of two essential immune functions (antigen processing and presentation, and T helper cell activation and tolerance), and to abnormalities in lipid metabolism

Edited by 100YearsToGo, 01 March 2009 - 04:46 PM.

[FunkOdyssey]

SIRT1: Regulation of longevity via autophagy.
Salminen A, Kaarniranta K.
Cell Signal. 2009 Feb 25. [Epub ahead of print]
PMID: 19249351

Recent studies have emphasized the importance of SIRT1, a mammalian homolog
of Sir2 longevity factor, in the regulation of metabolism, cellular
survival, and organismal lifespan. The signaling network interacting with
SIRT1 continues to expand as does the number of functions known to be
regulated by SIRT1. Autophagy is also an emerging field in longevity
studies. Autophagocytosis is a housekeeping mechanism cleaning cells from
aberrant and dysfunctional molecules and organelles. The extension of
lifespan has been linked to the efficient maintenance of autophagic
degradation, a process which declines during aging. Interestingly, recent
observations have demonstrated that SIRT1 regulates the formation of
autophagic vacuoles, either directly or indirectly through a downstream
signaling network. We will examine the signaling pathways linking SIRT1 to
the regulation of autophagic degradation. The interactions of SIRT1 with the
FoxO and p53 signaling can also regulate both the autophagic degradation and
lifespan extension emphasizing the key role of autophagy in the regulation
of lifespan.

Keywords: Aging; Caloric restriction; Lysosomes; p53; Sir2; Sirtuins.

Article Outline

1. Introduction
2. SIRT1 deacetylase: key regulator of metabolism and cell survival
3. SIRT1 and lifespan extension
4. Autophagy: lifespan regulation via housekeeping
5. SIRT1 and regulation of autophagy
5.1. SIRT1 regulates the assembly of Atg protein complexes
5.2. SIRT1 activators: activators of autophagy?
6. SIRT1 stimulates autophagy via a longevity factor network
7. Concluding remarks

[Mixter]

Another point, what exactly triggers mTOR mediated autophagy? Could the primary mTOR induction be a decrease of intracellular calcium?

Verapamil and lithium seem to be some of the most effective agents currently known. They both affect calcium homeostasis (verapamil decreases intracellular calcium, with lithium it seems more complicated, but it could ultimately release calcium from the cell too).

And indeed, I just found a confirmation: Testosterone Activates mTOR/S6K1 Pathway Through Intracellular Calcium and ERK in Cardiomyocytes

So, might one of the primary reason why autophagy is reduced in age be an increase in intracellular calcium? Might be interesting to find out, an obvious experiment could consist of inducing a reduction in accumulated intracellular calcium in aged animals and seeing whether autophagy activation is stronger after that.

[geddarkstorm]

Another point, what exactly triggers mTOR mediated autophagy? Could the primary mTOR induction be a decrease of intracellular calcium?

Verapamil and lithium seem to be some of the most effective agents currently known. They both affect calcium homeostasis (verapamil decreases intracellular calcium, with lithium it seems more complicated, but it could ultimately release calcium from the cell too).

And indeed, I just found a confirmation: Testosterone Activates mTOR/S6K1 Pathway Through Intracellular Calcium and ERK in Cardiomyocytes

So, might one of the primary reason why autophagy is reduced in age be an increase in intracellular calcium? Might be interesting to find out, an obvious experiment could consist of inducing a reduction in accumulated intracellular calcium in aged animals and seeing whether autophagy activation is stronger after that.

Apparently calcium actually activates macroautophagy. Also, calpain, calcium-dependent cysteine protease, is one of the controllers that modulate if a signal causes autophagy or apoptosis. In this case, calpain stops apoptosis and activates autophagy (what we want). So, calcium is very important for activating autophagy and preventing apoptosis through this effector.

Now, the interesting thing about calcium is that the signal it confers to the cell is dependent on its concentration. Low levels, medium levels, and high levels of calcium all mean different things to a cell. Very high levels of calcium influx to the mitochondria (as activated by certain signals) activates the mitochondrial transition pore, loss of mitochondrial coupling of proton motive force to ATP production, and massive increases in ROS production, which then signal and start apoptosis. Too low levels of calcium lead to cellular and mitochondrial metabolism dysfunction (seen in fasting mice by knocking out UCP3; which has been recently discovered to be the ruthenium red sensitive mitochondrial calcium uniporter, and not likely a direct uncoupler like UCP1). Medium levels of calcium (gained by a high calcium diet ironically) on the other hand stimulate ATP production and dehydrogenase activity, leading to obesity resistance and such, possibly through UCP3.

Add to this that old age is also marked by decreasing calcium content in the bones, and it seems the opposite is more likely: that the reason autophagy is reduced in old age is because of decreasing intracellular/intramitochondrial calcium levels.

The experiment you propose though should be easy and immediately answer this issue. Let's hope someone decides to do it just to see (if there isn't research I'm just not finding on it already).

Edited by geddarkstorm, 06 March 2009 - 02:22 AM.

[bkaz]

Very, very interesting. Hmm.. it sounds like humans don't naturally produce it ("No vertebrate has been shown to synthesize trehalose" ref), but also break it down very rapidly. That's a bummer. I wish I could find more in vivo kinetics data on it... maybe it lasts long enough in serum, and is taken up into and stored in cells enough to have an effect and justify use. This is definitely something to keep an eye on, thanks 100YearsToGo.

There must be a reason the vertebrates lost the capacity to produce Trehalose? Perhaps, as a chemical chaperone, it slows down functional interactions among proteins?

[geddarkstorm]

Very, very interesting. Hmm.. it sounds like humans don't naturally produce it ("No vertebrate has been shown to synthesize trehalose" ref), but also break it down very rapidly. That's a bummer. I wish I could find more in vivo kinetics data on it... maybe it lasts long enough in serum, and is taken up into and stored in cells enough to have an effect and justify use. This is definitely something to keep an eye on, thanks 100YearsToGo.

There must be a reason the vertebrates lost the capacity to produce Trehalose? Perhaps, as a chemical chaperone, it slows down functional interactions among proteins?

If that were so, we'd see such effects in experiments and tests. So far it seems pretty safe and advantageous, and is broken down just fine by the body.

Most likely vertebrates don't make it because there is no need. Doing so would be a loss of energy for too little gain. For instance, humans do not naturally make vitamin C, but goats and other vertebrates do. We, humans, have lost a lot of the metabolic capacity that even other higher vertebrates have. Vitamin C is obviously essential for us, so why don't we make it? Probably because we get enough of it in our food with our omnivore diet, so getting rid of its production saves on energy to make other things, like a big brain. That's all just my theory though, lol.

[bkaz]

There must be a reason the vertebrates lost the capacity to produce Trehalose? Perhaps, as a chemical chaperone, it slows down functional interactions among proteins?

If that were so, we'd see such effects in experiments and tests. So far it seems pretty safe and advantageous, and is broken down just fine by the body.

Most likely vertebrates don't make it because there is no need. Doing so would be a loss of energy for too little gain. For instance, humans do not naturally make vitamin C, but goats and other vertebrates do. We, humans, have lost a lot of the metabolic capacity that even other higher vertebrates have. Vitamin C is obviously essential for us, so why don't we make it? Probably because we get enough of it in our food with our omnivore diet, so getting rid of its production saves on energy to make other things, like a big brain. That's all just my theory though, lol.

Well, there're a lot of different interaction among proteins, we can't test for them all. We don't make vit.C because we used to to get plenty from the diet, but that's not the case with trehalose, especially since "it is broken down just fine by the body..". It would be very easy for for the vertebrates to preserve the ability to synthesize trehalose from starch if it was so "safe and advantageous"?

[geddarkstorm]

There must be a reason the vertebrates lost the capacity to produce Trehalose? Perhaps, as a chemical chaperone, it slows down functional interactions among proteins?

If that were so, we'd see such effects in experiments and tests. So far it seems pretty safe and advantageous, and is broken down just fine by the body.

Most likely vertebrates don't make it because there is no need. Doing so would be a loss of energy for too little gain. For instance, humans do not naturally make vitamin C, but goats and other vertebrates do. We, humans, have lost a lot of the metabolic capacity that even other higher vertebrates have. Vitamin C is obviously essential for us, so why don't we make it? Probably because we get enough of it in our food with our omnivore diet, so getting rid of its production saves on energy to make other things, like a big brain. That's all just my theory though, lol.

Well, there're a lot of different interaction among proteins, we can't test for them all. We don't make vit.C because we used to to get plenty from the diet, but that's not the case with trehalose, especially since "it is broken down just fine by the body..". It would be very easy for for the vertebrates to preserve the ability to synthesize trehalose from starch if it was so "safe and advantageous"?

Erm.. I think you missed my point? We can get it in our diet, considering plants, fungi, and insects make it (all sources of human food, yes, even insects haha; also why we have a metabolic enzyme specific for it). Just because we don't make it doesn't actually mean anything. It's like saying we don't need half of the 20 (actually 22) essential amino acids because we can't synthesize them anymore. Or even more relevant an example, look at bioflavanoids and how many beneficial effects they give -- only plants synthesize them, just like with trehalose, we don't synthesize them to conserve energy, not because they aren't useful.

And you can test for most every possible protein interaction that might be relevant. It's easy, just give trehalose to a rat or mouse at different concentrations and see what happens. Apparently, they do just fine, hence why it's been given GRAS (generally regarded as safe) status in the US and EU.

Edited by geddarkstorm, 09 March 2009 - 11:20 PM.