• @Blossom Is A Blessing To This Community, Let Us Be A Blessing To Her
    Click Here For More Information
  • Due to excessive bot signups along with nefarious actors we are limiting forum registration. Keep checking back for the register link to appear. Please do not send emails or have someone post to the forum asking for a signup link. Until the current climate changes we do not see a change of this policy. To join the forum you must have a compelling reason. Letting us know what skills/knowledge you will bring to the community along with the intent of your stay here will help in getting you approved.

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
Ray has written so many articles on how "autoimmune conditions" like MS, rheumatoid arthritis (RA), lupus, psoriasis, etc are all metabolic diseases mostly driven by estrogen. After decades of denial by mainstream medicine, a study was just published by a team at Stanford that shows RA to be indeed a metabolic disease. More importantly, RA was found to be a disease caused by excessive reductive stress caused by excessive metabolism of glucose through the pentose phosphate pathway (PPP), and accumulation of NAPDH and reduced glutathione, accompanied by reductions in oxidative phosphorylation. The PPP is a parallel pathway to glycolysis and exists primarily as a defense mechanism against the so-called reactive oxygen species (ROS). The excessive reductive stress apparently not only neutralizes the ROS but it also inhibits oxidative metabolism in the electron transport chain.

I read a review that gives a different take on NADPH. It puts a different light on NADPH as it is used actually in phagocytosis, where a respiratory (or oxidative) burst is needed to kill pathogens. It is actually used in creating ROS, as these are needed to kill pathogens. White blood cells are effective having the benefit of NADPH in creating ROS. Without ROS, white blood cells would be less effective and have to use other less effective means to deal with pathogens. With ROS, I get the impression less white blood cells are needed as they are very effective, armed with missile launchers, and without ROS, you need to have tons of white blood cells, equivalent to the Chinese hordes depicted in Korean War movies. This is going to create any remnants to form plaque, as well as a lot of inflammatory conditions.

This is the review: http://www.annclinlabsci.org/content/30/2/145.full.pdf

NADPH and the pentose phosphate pathway is scarcely mentioned in the forum, but its importance shouldn't be overlooked, as, among other things, it makes the sugars that are used to form nucleotides such as DNA and RNA. I found this series of blogs by an unknown blogger to be fascinating, regarding the PPP and thiamine:

Synergy Health & Wellness: The Pentose Phosphate Pathway: The missing link between hormonal imbalances and carbohydrate metabolism?
Synergy Health & Wellness: The importance of addressing thiamine status in adrenal fatigue
Synergy Health & Wellness: Redox balance, the pentose phosphate pathway, and adrenal function
Synergy Health & Wellness: NADPH, the folate cycle, and adrenal function
Synergy Health & Wellness: Acetylcholine and adrenal function: Is adrenal fatigue a lack of break fluid?
Synergy Health & Wellness: Thiamine, gut health, the immune system, and adrenal fatigue

The font and size makes the blog hard to read from my Chrome browser. I had to use the Mercury Reader extension to make it easier to read. I use Night Mode to make it easier on my eyes as well. Overall, it is well-organized and well-explained.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
Another possible therapeutic agent is methylene blue, since it can both oxidize NADPH and reduced glutathione AND also stimulate mitochondrial activity and thus the generation of ROS.
I am puzzled about this statement. While oxidizing GSH would help with generating ROS, I don't understand why oxidizing NADPH would. Isn't NADPH oxidase needed to produce ROS? So how would not having NADPH available be helpful in producing ROS? Anyone?
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
I am puzzled about this statement. While oxidizing GSH would help with generating ROS, I don't understand why oxidizing NADPH would. Isn't NADPH oxidase needed to produce ROS? So how would not having NADPH available be helpful in producing ROS? Anyone?
I am less puzzled now as @haidut made a typo here and did not correct it. It's only one letter - "reduced" should be "reduce" such that the statement should read:

Another possible therapeutic agent is methylene blue, since it can both oxidize NADPH and reduce glutathione AND also stimulate mitochondrial activity and thus the generation of ROS.

But I am still confused by this statement.

If NADPH is oxidized, and glutathione is reduced,

-this means there is less NADPH oxidase, and with NADPH oxidase being an enzyme needed to generate ROS, how can this be helpful in generating ROS?
-this means with more GSH, less ROS is produced because GSH counters the production of ROS, so how can this help towards ROS generation?

And while methylene blue stimulates mitochondrial activity, it does not necessarily lead to an increase in the generation of ROS. On the other hand, use of methylene blue can lead to less ROS generation during mitochondrial respiration. This is as I understand it watching these two videos by Dr. Francisco Gonzales Lima:




which I watched thanks to @R J on his thread: Informative Interview About Methylene Blue

@Hans @redsun @zarrin77 How am I off here?
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
I am less puzzled now as @haidut made a typo here and did not correct it. It's only one letter - "reduced" should be "reduce" such that the statement should read:

Another possible therapeutic agent is methylene blue, since it can both oxidize NADPH and reduce glutathione AND also stimulate mitochondrial activity and thus the generation of ROS.

But I am still confused by this statement.

If NADPH is oxidized, and glutathione is reduced,

-this means there is less NADPH oxidase, and with NADPH oxidase being an enzyme needed to generate ROS, how can this be helpful in generating ROS?
-this means with more GSH, less ROS is produced because GSH counters the production of ROS, so how can this help towards ROS generation?

And while methylene blue stimulates mitochondrial activity, it does not necessarily lead to an increase in the generation of ROS. On the other hand, use of methylene blue can lead to less ROS generation during mitochondrial respiration. This is as I understand it watching these two videos by Dr. Francisco Gonzales Lima:




which I watched thanks to @R J on his thread: Informative Interview About Methylene Blue

@Hans @redsun @zarrin77 How am I off here?

MB lowers superoxide production, but increases H2O2 production, reducing the superoxide:H2O2 balance. Superoxide is thought to be pro-cancer, whereas H2O2 is anti-cancer. I'm in the process of writing a big article about this which should be out soon I hope.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
MB lowers superoxide production, but increases H2O2 production, reducing the superoxide:H2O2 balance. Superoxide is thought to be pro-cancer, whereas H2O2 is anti-cancer. I'm in the process of writing a big article about this which should be out soon I hope.
Thanks Hans on elucudating on the role of methylene blue.

In the question of what happen when NADPH is oxidized at the same time as glutathione is reduced, doesn't this lead to a condition that does not favor ROS production, being that there is no (or less) NADPH oxidase (NOX) to enable the production of ROS, and the increased availability of GSH neutralizes the oxidative action of ROS. This would not favor the beneficial use of ROS in the respiratory burst of phagocytosis aimed at destroying pathogens, as part of the body's immunity?

Looking forward to your article.
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
Thanks Hans on elucudating on the role of methylene blue.

In the question of what happen when NADPH is oxidized at the same time as glutathione is reduced, doesn't this lead to a condition that does not favor ROS production, being that there is no (or less) NADPH oxidase (NOX) to enable the production of ROS, and the increased availability of GSH neutralizes the oxidative action of ROS. This would not favor the beneficial use of ROS in the respiratory burst of phagocytosis aimed at destroying pathogens, as part of the body's immunity?

Looking forward to your article.
NADPH is used by NOX and glutathione reductase (GR) to produce ROS and quench ROS. It's a balancing act. Methylene blue is reduced by NADPH, so methylene blue lowers total NADPH, which could reduce NOX and GR activity. In vitro evidence show that methylene blue inhibits NOX, but excess NOX is the cause of degeneration.
On the other hand, methylene blue increases H2O2, which is able to enhance macrophage phagocytic activity.
Besides, methylene blue protects against viral infections, so I doubt that methylene blue would prevent your body from doing proper phagocytosis.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
NADPH is used by NOX and glutathione reductase (GR) to produce ROS and quench ROS. It's a balancing act. Methylene blue is reduced by NADPH, so methylene blue lowers total NADPH, which could reduce NOX and GR activity. In vitro evidence show that methylene blue inhibits NOX, but excess NOX is the cause of degeneration.
On the other hand, methylene blue increases H2O2, which is able to enhance macrophage phagocytic activity.
Besides, methylene blue protects against viral infections, so I doubt that methylene blue would prevent your body from doing proper phagocytosis.

I don't doubt the beneficial effects of methylene blue.

I question the underlying thought that reducing NADPH is beneficial and that increasing GSH is always good. As I had explained NADPH is needed in NADPH oxidase, an enzyme needed to make ROS, without which ROS cannot be produced. Being that ROS is employed in ther respiratory burst of phagocytosis, it is a crucial component of our immune system.

There are also times when too much GSH is not good, as GSH neutralizes ROS.

And so yes, there is a balancing act and a body in homeostatis will know how to balance the use of oxidants and reductants (antioxidants) to its advantage. If we are going to assist the body, it helps to know what we take actually does, in a properly understood context, so that we help the body instead of obstruct, in the way it responds to insults and in so doing the body heals and maintains homeostasis and health.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
The pentose phosphate pathway (PPP) is where, among other things, NADPH is produced, using sugar. In a way, it competes with the OxPhos pathway. When it is very active, it would take away resources from the OxPhos pathway. Seen this way, it is miscast as the evil twin of OxPhos. Some would say NADPH is used to make fats, so it is bad. Some would say it is used to make ROS, so it is bad. So bad that it would help to downregulate the PPP.

But little is said about the PPP. So it is misunderstood. It is actually the neglected twin of OxPhos. I feel sorry for it that I should stand up and defend it, and be its advocate. The truth of the matter is that the PPP doesn't demand for ROS to be produced. It merely acts as a helper to produce ROS. When the body needs ROS to be produced, it makes sure it keeps the protein NADPH to be readily avaiable to produce the enzyme, NADPH oxidase, to produce ROS. And ROS is what's needed to produce the respiratory oxidative burst on pathogens by phagocytes, such as neutrophils and macrophages, to destroy these pathogens. So what is so bad about ROS? What is so bad about NADPH?

If there are few pathogens in the body, there would be less ROS produced, and it would be logical to think that the PPP would have less need for NADPH to be produced. So it makes sense that in a body less exposed to chronic infection, there would be less NADPH produced - because there is less need for it.

But it is a bad twist of logic to think that by merely reducing the production of NADPH, or by reducing NADPH by oxidizing it, that the same body would become healthier. Because the absence of NADPH would mean the pathogens are not being destroyed by ROS. NADPH production is high because it is needed by the body. It's not because NADPH is produced in large amounts by a runaway process that needs to be controlled.

I think that if we can fix the chronic infections effectively, we can exert less demand on the PPP, and when the PPP is freed from such demands, the use of resources would flow back to the OxPhos, and with this, we can produce more energy. This can be experienced by higher heart rate, with more energy surplus left over for regeneration and development. There is less aging as well.

But if our body has many issues, the OxPhos has to downregulate as the PPP is needed more to deal with the issues. Survival is the priority over anti-aging.
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
I don't doubt the beneficial effects of methylene blue.

I question the underlying thought that reducing NADPH is beneficial and that increasing GSH is always good. As I had explained NADPH is needed in NADPH oxidase, an enzyme needed to make ROS, without which ROS cannot be produced. Being that ROS is employed in ther respiratory burst of phagocytosis, it is a crucial component of our immune system.

There are also times when too much GSH is not good, as GSH neutralizes ROS.

And so yes, there is a balancing act and a body in homeostatis will know how to balance the use of oxidants and reductants (antioxidants) to its advantage. If we are going to assist the body, it helps to know what we take actually does, in a properly understood context, so that we help the body instead of obstruct, in the way it responds to insults and in so doing the body heals and maintains homeostasis and health.
There are some NOX enzymes that don't use NADPH actually. Plus, if there is higher amounts of H2O2 in the body, then the NOX enzymes don't have to work as hard to produce superoxide, as the H2O2 is helping a lot to keep things in check.
If NADPH drops, then some of the NOX activity drops, but so does GSH levels, so even if less superoxide is being produced, less if quenched by GSH, so in theory, the effect is still the same.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
There are some NOX enzymes that don't use NADPH actually.
How can this be? NOX means NADPH oxidase. How can NOX enzymes not use NADPH?

Plus, if there is higher amounts of H2O2 in the body, then the NOX enzymes don't have to work as hard to produce superoxide, as the H2O2 is helping a lot to keep things in check.
I'm not so sure about this. But yes, hydrogen peroxide is not as harmful as superoxide radical anion, as it is not really a free radical. But in the production of thyroid hormones, too much hydrogen peroxide can destroy the thyroid gland tissues, and that's why glutathione peroxidase is needed to convert hydrogen peroxide to water.

If NADPH drops, then some of the NOX activity drops, but so does GSH levels, so even if less superoxide is being produced, less if quenched by GSH, so in theory, the effect is still the same.
It's not that simple. If NADPH production is lacking, enzymes needed to make ROS for phagocytosis will be unavailable, and this will compromise the immune system's functions in destroying pathogens. If NADPH is sufficient, it will be available for use. It doesn't have to be used if it's not needed. For example, it there is enough cytochrome c oxidase to donate electrons to oxygen in oxphos, ROS production will be minimized in the course of oxphos. In this situation, less glutathione will be needed to neutralize ROS, and less NADPH will be needed to recycle oxidized glutathione (GSSH) to reduced glutathione (GSSH).
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
How can this be? NOX means NADPH oxidase. How can NOX enzymes not use NADPH?
I read a study where DUOX 1 and 2 didn't require NADPH, but now I can't find it again.
I'm not so sure about this. But yes, hydrogen peroxide is not as harmful as superoxide radical anion, as it is not really a free radical. But in the production of thyroid hormones, too much hydrogen peroxide can destroy the thyroid gland tissues, and that's why glutathione peroxidase is needed to convert hydrogen peroxide to water.
H2O2 potentiates the effects of superoxide at phagocytosis, thus theoretically reducing superoxide requirements. It's like having a spotter. If someone is helping you, you don't need to work as hard.
If NADPH production is lacking, enzymes needed to make ROS for phagocytosis will be unavailable, and this will compromise the immune system's functions in destroying pathogens.
Yes, but we started talking about this because the fact that methylene blue lowered NADPH. There is no evidence that methylene blue depletes NADPH or hamper proper phagocytosis. Perhaps something else that tanked NADPH would hinder the body to fight against pathogens, but I'm sure methylene blue doesn't.
For example, it there is enough cytochrome c oxidase to donate electrons to oxygen in oxphos, ROS production will be minimized in the course of oxphos. In this situation, less glutathione will be needed to neutralize ROS, and less NADPH will be needed to recycle oxidized glutathione (GSSH) to reduced glutathione (GSSH).
Agreed. Also, the body will only make more NADPH when really needed. For example, ROS inhibits pyruvate kinase M2, which when shuttles glucose down the PPP, instead of oxidative phosphorylation. More NADPH is made and GSH is created to quench the ROS.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
I read a study where DUOX 1 and 2 didn't require NADPH, but now I can't find it again.
When you find it, please share.
H2O2 potentiates the effects of superoxide at phagocytosis, thus theoretically reducing superoxide requirements. It's like having a spotter. If someone is helping you, you don't need to work as hard.
Interesting. Hope you can expand on this in your upcoming article.
Yes, but we started talking about this because the fact that methylene blue lowered NADPH. There is no evidence that methylene blue depletes NADPH or hamper proper phagocytosis. Perhaps something else that tanked NADPH would hinder the body to fight against pathogens, but I'm sure methylene blue doesn't.
Yes, methylene blue doesn't deplete NADPH. It may actually conserve NADPH. Methylene blue donates electrons to oxidized cytochrome c oxidase to keep cyt c oxidase from becoming exhausted in donating electrons to oxygen, thereby minimizing oxygen from turning into a superoxide radical during oxphos. I've read that generally 10% of the oxygen involved in oxphos turns into superoxide radicals during oxphos, but if that can be reduced significantly then it means the body won't have to produce antioxidants to counter that oxidative stress. The less the need to produce antioxidants, the less the need to produce NADPH to recycle oxidized glutathione (GSSG) back to reduced glutathione (GSH).

For example, ROS inhibits pyruvate kinase M2, which when shuttles glucose down the PPP, instead of oxidative phosphorylation. More NADPH is made and GSH is created to quench the ROS.
I want to test my understanding of this. So pyruvate kinase M2 inhibits glucose from the PPP then? So that when ROS inhibits pyruvate kinase M2, it allows or modulates glucose shuttling to the PPP, such that more NADPH can be produced to enable glutathione recycling to have GSH available to quench ROS?
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
I want to test my understanding of this. So pyruvate kinase M2 inhibits glucose from the PPP then? So that when ROS inhibits pyruvate kinase M2, it allows or modulates glucose shuttling to the PPP, such that more NADPH can be produced to enable glutathione recycling to have GSH available to quench ROS?
Pyruvate kinase M2 (PKM2) is the last enzyme in glycolysis which creates pyruvate. If PKM2 is overactive, a lot of lactate will be produced. When PKM2 is inhibited, the glucose has to go somewhere else, and that will be through the PPP. Beta-lapachone inhibits excess PKM2 in cancer thus lowering lactate.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
Pyruvate kinase M2 (PKM2) is the last enzyme in glycolysis which creates pyruvate. If PKM2 is overactive, a lot of lactate will be produced. When PKM2 is inhibited, the glucose has to go somewhere else, and that will be through the PPP. Beta-lapachone inhibits excess PKM2 in cancer thus lowering lactate.

From how you phrased it, it seems like a choice of having too much lactate or NADPH through the PPP. But I think it's more like a choice of more OxPhos or more PPP. With more ROS production, more NADPH needs to be produced to enable production of antioxidants like GSH to neutralize the ROS. And to enable NADPH production, PPP needs to be up-regulated and OxPhos needs to be down-regulated. And inhibiting PKM2 downregulated OxPhos, and up-regulates PPP.
 

Hans

Member
Joined
Aug 24, 2017
Messages
4,810
From how you phrased it, it seems like a choice of having too much lactate or NADPH through the PPP. But I think it's more like a choice of more OxPhos or more PPP. With more ROS production, more NADPH needs to be produced to enable production of antioxidants like GSH to neutralize the ROS. And to enable NADPH production, PPP needs to be up-regulated and OxPhos needs to be down-regulated. And inhibiting PKM2 downregulated OxPhos, and up-regulates PPP.
Yes that's how I understand it as well, because excess ROS also inhibits PDH. So perhaps PDH is inhibited first by a certain level of ROS and if the ROS goes even higher, then PKM2 gets inhibited as well.
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
Yes that's how I understand it as well, because excess ROS also inhibits PDH. So perhaps PDH is inhibited first by a certain level of ROS and if the ROS goes even higher, then PKM2 gets inhibited as well.

As I get to understand more how our body works, the more I see how it knows how to balance itself with what it's got. If I provide it with good nutrition, it is on autopilot to arrive at optimality. And if it's not reaching an optimal state, more support may be needed in terms of nutrients and substances. But mainly essential ones, which are exogenous.

One example I think of is thiamine. If more NADPH needs to be produced, then increased thiamine intake is needed. Otherwise, thiamine may become the limiting factor, and with NADPH running out, phagocytosis as well as antioxidants would be in short supply.
 

Mito

Member
Joined
Dec 10, 2016
Messages
2,484
As I get to understand more how our body works, the more I see how it knows how to balance itself with what it's got. If I provide it with good nutrition, it is on autopilot to arrive at optimality.
Do you think aging impacts the body’s ability to balance itself with good nutrition alone?
 

yerrag

Member
Joined
Mar 29, 2016
Messages
9,866
Location
Manila
Do you think aging impacts the body’s ability to balance itself with good nutrition alone?

It's a matter of degree. Everything being the same, as one gets older, the stronger the effect of age. But if the body is in good shape, having optimal metabolism and energy from good nutrition, and not suffering from chronic inflammation from pathogens, toxins, and auto-immunity (an adaptation - to meet the present need to survive while trading the future for it, while throttling energy to mitigate damage) the body would not be aging as much. And if the body isn't aging as much, it will continue to have that balance.
 

Similar threads

Top