[Study] Acquired Mitochondrial Abnormalities Including Epigenetic Inhibition of Superoxide Dismutase

[Learner1]

I apologize right up front for bringing up something you might consider a closed issue. It looks like LPS can cause an increase in iNOS, peroxynitrite and superoxide-
LINK
If you do have higher than normal levels of LPS, that could explain the high ROS levels and why the massive doses of antioxidants you are taking can't bring them down. Because the cause is still there, to some degree, constantly creating more ROS? A possibility maybe?

Martin Pall and Maes and Morris seemed to believe there were other things driving this.

But, it's worth investigating. Exactly how does one figure out if one has this problem? I've done multiple brands of stool tests and haven't seen it come up.

 

[Learner1]

You might be interested in the following recapture, including ADP and misleading electrons [my translation, and I am still lazy... ]
"Oxidativer Stress - ein Risikofaktor für mitochondriale Deletionen." (internet title I think); Franziska Weber:
"Die Rolle von oxidativem Stress bei der Entstehung von Deletionen der mitochondrialen DNA ..." diss 2011.

"1.322 ... The reduction O2 -> H2O is bound to the phosphorylation of ADP via DeltaPsi [ie electric gradient]. When the exogenous level of ADP is huge, the ATP synthase is driven by complex V, DeltaPsi is becoming wider and O2 is used for the purpose to rebuild DeltaPsi (state III-respiratory). When the level of ADP is a restriction, DeltaPsi is becoming less, ETC slows down, and O2 is used only low (state IV-respiratory). The ratio III : IV is called respiratory control ratio (RCR) (Kokosza JE, 2001).
0.4-4% of the moelcular O2 used up by mt is resulting in a toxic byreaction: Out of the initial steps of the ETC the high toxic {O2->} is built instead of H2O via a transfer of a single electron. The main source of such electrons, which are building {O2->}, is complex I (Chance B 79, Turrens JF 85, Murphy MP 09). The SOD2 makes {O2->} becoming H2O2. The mitochondrial H2O2 is then reduced to H2O by the Gluthationperoxidase-1 (Gpx-1). But in appearance of reduced transition metals the H2O2 can get converted to high toxic {OH.} via the Fenton-reaction (Giulivi C 95). When the OXPHOS is hemmed the rate of mischieving electrons elevates and therefore ROS. That means, in the passiv [?] state (=IV) the ROS production is higher than in the activated state (=III)."


The next sentences in short: to high for detox capacitiy -> swollen mt -> cytochrome c gets into the plasma -> apoptosis.
The goal of the diss was to show a causing effect on aging, but couldn´t pro nor con.
Learner1, you might be interested in the overview on page 32, guessing should not be difficult mostly.

Looks great - I'd love to read it, but 104 pages in German is a little much for me. How did you get the English translation?

 

[nanonug]

I've done multiple brands of stool tests and haven't seen it come up.

LPS (lipopolysaccharide) is something that you'd measure in your blood. Don't know which labs offer this test, though.

 

[Learner1]

@Leaner1 First off I will say I don't know if reducing oxidative stress is a good idea or a bad idea. I would guess that in a CFS patient who is producing lots chronically, lowering it could be a good thing to get the body "back in balance".

I think there's a lot to be said for normalizing various processes in the body. The more we function normally, the more likely our bodies are to act normally.

What I mean is that it could well be that acute inflammation is necessary and good, yet chronic creates a vicious circle which needs to be broke out of.

The interesting thing for me is my inflammation markers are quite low.

Yet I have come across the idea that inflammation isn't bad several times now from several different sources.

Agreed.

Inflammation is the first step in the repair process after some physical or chemical injury or stress. It destroys microbes to eradicate infections and prevents the spread of damaged cells to other areas of the body that could cause secondary problems, and it rids the body of dead and damaged cells. It is a part of ridding the body of toxins.

The reverse electron flow superoxide generation seems to be very important in insulin signalling and energy status signalling. ROS seem to be essential for the immune system to fight pathogens.

But other complexes create ROS also.

Robert O'Becker showed that inflammation at an injury site was a signal to direct healing efforts to that area. [Brain + spine = positive charge, extremeties negative charge. Cut off salamanda leg and the wound becomes positively charged (inflammation) which directs stem cells/nutrients/immune/etc to that area; limb regenerates. If you get rid of inflammation / positive charge in this area, limb does not regenerate.

I am also listening to a chris masterjohn podcast right now were he is "ranting" about how antiinflammatories aren't necessarily good and interfere with signaling.

There is a lot of complexity, as you're pointing out. And multiple chemicals are signalling. Calcium and acetylcholine signalling are involved in all this too.

I think our best bet is trying to support the body in what it's doing. If glutathione or sod are overwhelmed then do things to support them as this will probably lead to excess oxidation than is needed for signaling and cause damage.

Agreed. This is what both my ME/CFS doctor and my functional medicine doctor are trying to do. However, it seems like a huge order.

Recently, after dealing with my infections, upping my amino acids as I'm burning them (per the Fluge and Mella PDH study) and upping my NAD+, I've been able to increase exercise to normal levels. However, I've gotten slammed with PEM. First, BCAAs helped. But I'm needing reduced glutathione, too. Doing both heads off or rescues me from the PEM.

I think maybe we are producing all this inflammatory signalling because of an underlying chronic infection but I really have no idea.

After 6 months of IVIG, 10 months of valganciclovir and LDN, and 4 months of IV antibiotics, I'd like to think my 7 chronic infections are under control. I certainly feel much better and can do more...

I intend to try certain anti inflammatory agents etc, and see if I feel better or not. This is better than relying too much on theories I guess.

I've been on curcumin the whole time, which may be helping my inflammation, because u don't seem to have too much.

Somehow, I think my body is topped into some state that's sucking up resources and causing all the free radicals. My labs seem to support this and supplementing to respond to the various stressors helps.

The metabolomics and other ME/CFS research seems to have found a lot of these sane abnormalities, so I suspect this is going on in others, too. I feel fortunate to have such a plethora of lab work to have identified all of this and to have has such good doctors solving my problems, one by one, but its clear this is still very complex and there's something snarky in our systems underlying all of this.

The question is how to kick the body out of this state?

 

[Learner1]

LPS (lipopolysaccharide) is something that you'd measure in your blood. Don't know which labs offer this test, though.

Yes, I can see that from the studies. But I don't see any lab tests popping up, so I have no idea how one can identify it or fix it.

 

[ljimbo423]

Martin Pall and Maes and Morris seemed to believe there were other things driving this.

But, it's worth investigating. Exactly how does one figure out if one has this problem? I've done multiple brands of stool tests and haven't seen it come up.

I agree with Martin Pall and Maes theories to some degree. I think mito dysfunction can cause high levels of ROS and it can also cause immune system dysfunction.

Creating to some degree a feedforward self-sustaining cycle. I do think that something has to be driving it, for it to be sustained though or high enough doses of antioxidants etc, would stop it.

How my recent coq10 increase to 500 mg a day stopped the horrendous flu-like flares I was getting, that would put me in bed for days, is incredible! That has pretty much convinced me that mito dysfunction is causing my flu-like symptoms, by somehow altering my immune system. The coq10 has stopped that process.

Maes talks about different ongoing triggers to his theory, bacterial translocation being one-

The proposal made herein is that once generated chronically activated O&NS
and immune-inflammatory pathways conspire to generate a multitude of self-sustaining and self-amplifying pathological
processes which are associated with the onset of ME/CFS. Sources of continuous activation of O&NS and immuneinflammatory
pathways in ME/CFS are chronic, intermittent and opportunistic infections, bacterial translocation,
autoimmune responses, mitochondrial dysfunctions, activation of the Toll-Like Receptor Radical Cycle, and decreased
antioxidant levels.

LINK

I agree with @nanonug, I think a blood test is how to test for it but also don't know where to get the test.

 

[ljimbo423]

Maes also says that "intermittent and opportunistic infections" could be sustaining the cycle. If that's true, maybe exercise causes an increase in LPS into the bloodstream and that's enough for some, to maintain the cycle.

One study showed a sustained increase in bacterial DNA for 72 hours, in the blood of CFS patients, after an exercise challenge, over controls.

 

[Learner1]

From the Maes quote above "there are autoimmune responses, mitochondrial dysfunctions, activation of the Toll-Like Receptor Radical Cycle."

I suspect the Toll-Liker Receptor Radical Cycle has a good deal to do with our issues...

CNS Neurol Disord Drug Targets. 2015;14(7):838-54.
The Toll-Like Receptor Radical Cycle Pathway: A New Drug Target in Immune-Related Chronic Fatigue.
Lucas K, Morris G, Anderson G, Maes M

Abstract

In this review we discuss that peripheral and central activation of the Toll-like receptor 2/4 (TLR2/4) Radical Cycle may underpin the pathophysiology of immune-related chronic fatigue secondary to other medical diseases and conditions. The TLR Radical Cycle plays a role in illnesses and conditions that are disproportionately commonly comorbid with secondary chronic fatigue, including

a) neuroinflammatory disorders, e.g. Parkinson's disease, stroke, depression, psychological stressors, and

b) systemic disorders, e.g. (auto)immune disorders, chronic obstructive pulmonary disease, ankylosing spondylitis, chronic kidney disease, inflammatory bowel disease, cardiovascular disease, incl. myocardial infarction, cancer and its treatments.

Increased TLR signaling is driven by activated immuneinflammatory and oxidative and nitrosative stress pathways, pathogen derived molecular patterns, including lipopolysaccharides, and damage associated molecular patterns (DAMPs). Newly formed redox-derived DAMPs, secondary to oxidative processes, may further activate the TLR complex leading to an auto-amplifying TLR Radical feedback loop.

Increased gut permeability with translocation of gram negative bacteria and LPS, which activates the TLR Radical Cycle, is another pathway that may play a role in most of the abovementioned diseases and the secondary fatigue accompanying them.

It is concluded that secondary fatigue may be associated with activation of the TLR Radical Cycle pathway due to activated immune-inflammatory pathways, classical and redox-derived DAMPs and PAMPs plays a role in its pathophysiology. Such an activation of the TLR Radical Cycle pathway may also explain why the abovementioned conditions are primed for an increased expression of secondary chronic fatigue.

Targeting the TLR Radical Cycle pathway may be an effective method to treat TLR-Radical Cycle-related diseases such as secondary chronic fatigue.

This is an illustration of things affecting the TLR Radical Cycle pathway. It also includes heavy metals and other cell dangers...

This paper discusses Toll Like Receptors and how they promote inflammation and disease, which is likely very important for many of us:

https://www.nature.com/articles/emm201328#t2

Regulatory checkpoints in the Toll-like receptor (TLR) pathway. Subsequent to stimulation with their cognate ligands, TLRs induce several mediators converging at nuclear factor-κB (NF-κB), which mobilizes the transcription of inflammatory genes. This helps the host prepare to fight the microbial threat. After eliminating the threat, the cell must turn off the expression of inflammatory mediators. Sustained activation and overactivation of TLRs are usually accompanied by deleterious consequences for the host.

To prevent these consequences, the cell employs a variety of mechanisms to regulate TLR signaling, including decoy factors, adaptor modification, ubiquitin-mediated degradation, promoter state alteration, and translation disruption. All these mechanisms, individually or in combination, exert the effect of controlling inflammation and supporting the system to regain its normal state (arrows in red indicate the irreversible regulation, whereas the others show the reversible regulation).

If the immune system does not respond appropriately during a bacterial challenge, septic shock occurs, leading to multiple organ failure and death of the host. Sustained activation and overactivation of TLRs also results in the pathogenesis of autoimmune, chronic inflammatory, and neoplastic diseases. Therefore, pro- and anti-inflammatory responses must be balanced. Treatment with soluble receptors is one such physiological approach for modulating the immune response.
...
These studies have unveiled the role of TLRs in inflammation as well as the undesired roles of TLRs in several inflammatory diseases. Overactivation of TLRs disrupts homeostasis and places the patient at an increased risk of developing autoimmune diseases. To control hyper-responses, cells have evolved different mechanisms to protect the host, involving a plethora of molecules. At different stages, different molecules prevent the excessive secretion of inflammatory mediators and assist the immune system in restoring homeostasis. In the absence of these regulatory molecules, fatal consequences are inevitable

...
During homeostasis, when pathogen/damage-associated molecular patterns (PAMPs/DAMPs) breach the protective barrier, TLRs sense these patterns and mount an inflammatory response. As part of the inflammatory response, TLRs activate negative regulators, which have also shown to be triggered by inflammatory cytokines. The inflammatory response neutralizes the danger, and negative regulators prevent the overactivation of the immune system to protect the host. In the case of lack of appropriate TLR stimulation, the propagation of signals, or the induction of an inflammatory mediator, bacteremia may occur, whereas overactivation of any component leads to the development of inflammatory diseases and septic shock.

 

[percyval577]

[referring eg to Pall´s cycle, 17.4.]
I want to emphasize that NO instead of O2- could be the issue if NO is to much: NO + OO- -> ONOO-
(but Learner1 seems to be low on NO).

Jin, Kanthasamy et al 2007: "Mitochondria-targeted antioxidants for treatment of Parkinson's disease: Preclinical and clinical outcomes":
"Because of its negative charge and poor membrane permeability, superoxide is relatively unreactive, but it can react rapidly with nitric oxide (NO) to form the potent oxidant and nitrating agent peroxynitrite (ONOO−) and subsequently other reactive nitrogen species (RNS). Moreover, it is able to damage some mitochondrial iron–sulfur cluster-containing proteins [33]."

@Leaner1 First off I will say I don't know if reducing oxidative stress is a good idea or a bad idea.

I think it´s unimportant enough (if you eat some VitC and E). I even mean to remember that the theory of huge detrimentral effects (under normal conditions) has been shown to be wrong, but I might have dreamed so and I wouldn´t insist.

(percyval577): "you have the choice:
you can try (a) to diminish the O-molecules and to elevate the N-molecules (by elevating manganese)
or you can try (b) to elevate the O-molecules and to diminish the N-molecules (by lowering manganese)."

Wasn't aware of this, thanks for pointing it out. I'll look into this further although I'm supplementing at or close to RDA levels so I suspect it shouldn't be a problem.

Thanks for the thanks! I doubt that the RDA is aware of this alternation, and therefore should not be trustful here. I even think that nobody so far has come across this. I know, I might be an idiot, but even an idiot can say something true.
I remember well and never agreed: "Don´t eat much cholesterine." It´s guessing around without much sense and it takes time to get some clue. Now it turns out to have been nonsense. "Don´t loose track to your body, he wants to tell." my grandmother told me.

How did you get the English translation?

It´s once more my grandmother, she rather couldn´t speak German, and now my German turns automatically in other languages (but mostly not proper). I am from northern germany.

 

[Learner1]

I want to emphasize that NO instead of ROS could be the issue if NO is to much and would react with superoxide.

Very true. I'm trying to determine, though why my NO is low, at least some of the time. My doctor and I suspect its being used up to make ONOO-.

I found this in the attached by Martin Pall:

BH4 is a cofactor in nitric oxide synthases, such that when these NOSs are missing BH4, they become uncoupled, producing superoxide in place of nitric oxide. In partial uncoupling this superoxide can react, in turn, with the nitric oxide produced by adjacent coupled enzymes, leading to more peroxynitrite. Because the reaction between superoxide and nitric oxide is extraordinarily rapid, what is called diffusion controlled, the
production of both molecules by adjacent enzymes may be particularly effective in
raising peroxynitrite levels. Thus, although this partial uncoupling lowers nitric oxide
production, it is expected to increase peroxynitrite production, the most central part of the wider NO/ONOO- cycle

Given this, I suspect I need to focus more on BH4 production and recycling, and possibly take the BH4 precursors biopterin and sapropterin.

 

[Learner1]

A little research... Sapropterin is the drug Kuvan which is a "specialty medication" and exorbitantly expensive.

Biopterin used to be easy to get, but due to the Sapropterin patent, it is difficult to find in any reasonable dose.

And, rather than reinvent the wheel... these have been discussed before...

http://forums.phoenixrising.me/inde...ent-of-mitochondrial-replication-in-cfs.9536/

http://forums.phoenixrising.me/index.php?threads/biopterin-supplement.22788/

 

[percyval577]

I found this in the attached by Martin Pall:

I havn´t digged much into his theory. I recall: "Central to this theory are six positive feedback loops, which act such that elevated peroxynitrite increases the levels of both, nitric oxide and superoxide, reacting to more peroxynitrite." 2001

But then the production of NO [must slow down]
due to the increasing depletion of BH4
(a cofactor for the NO-synthases, which ones do not act anymore)
and now additional superoxide is produced - instead of nitric oxide

but there are "adjacent coupled [NOS] enzymes"
and this then "is expected to increase peroxynitrite,
the most central part of the wider NO/ONOO cycle"
​

(attachement from Learner1). This is how I understand it so far.

The wider cycle then includes furthermore: PRN->​

• ATP is going down-> NMDA is going up-> Ca is going up-> NF-kB is going up
• oxidative stress is going up-> Ca is going up
• superoxide is going up-> TRP rec is going up-> NMDA and Ca are going up

PRN yields directly NF-kB, which interestingly not only acts on the iNOS enzyme
but also - as far as I know since yesterday - would act on mt-biogenesis
(nevertheless I am not keen of a lot of NF-kB).​

The theory relies mainly on clinical improvements if I am right.

 

[Learner1]

This is all what I am understanding, too, except its more than just clinical observation.

Things that can be measured are nitric oxide, nitrotyrosine, vitamin C, folate, and B12.

Any others?

 

[Learner1]

I've done a bit of digging. It seems that sapropterin got patented and made into s medication named Kuvan, which is designated as an orphan drug and handled only by specialty pharmacies.(Read...very expensive. Like $3-4,000 a month, if one qualifies.)

I tried seeing about compounding, but I don't think it is allowed.

To read more abkut BH4 deficiency and treatment, click here:

https://emedicine.medscape.com/article/949470-overview#a5

The on-label use for Kuvan is for the genetic defect PKU, and there are a few other genetic defects that qualify.

Anyone know of other ways to get it or biopterin these days?

 

[knackers323]

I'd love to try some kuvan but irc.bio sells BH4 and apparently reputable

 

[Learner1]

I'd love to try some kuvan but irc.bio sells BH4 and apparently reputable

Sounds great. The price is, too. However one must have an "approved account" to buy, with these qualifications:

Qualifying clients are experienced researchers interested in the following:

• Life science, biochemistry, and neuroscience research
• Bulk quantity and exclusive product offers
• Long term cooperation

Clients who meet these criteria will be eligible to apply to our account approval program.

Not sure a regular customer can qualify...can they?

 

[percyval577]

Any others?

Would be nice though. (Maybe heam-CO? probably not very convincing even if true, NMDA/NO-AGM/CO? my guess)

I'm trying to determine, though why my NO is low, at least some of the time. My doctor and I suspect its being used up to make ONOO-.

Your physical constitution seemed to have some silly need for all that nitric oxide. The onliest method to find out might be to test some intakes/levels ratios. (bloodlevels or whatever)

I ever wondered if a low NO fatique would be thinkable. I myself know that too much nitric oxide is my problem:
Under certain circumstances I got/get good effects from inhibiters, and two times out of two I felt worse from arginine (onliest precursor of NO).

If you ask me by which entities the need would be driven my guess would be: epigenetics.
They would make that all that NO, and whatever, is taken up to great extent and rapidly.
And only if you would stay away from eating for some time the silly consequences would slow down
(which is reported by some of us when they feel better without eating for some time).

All this then would be thinkable for any possible relevant stuff
but everybody may feel free to think my manganese issue would be rubbish.
(I have a second guess with nickel that would not match up with your mt-complexes).

Anyhow, with almost any Mn-SOD knockout you wouldn´t be able to live, wether its direct, indirect or both a path.

 

[Learner1]

I ever wondered if a low NO fatique would be thinkable. I myself know that too much nitric oxide is my problem:

Have you measured it? My doctor is having me use Humann test strips, available on Amazon.

Under certain circumstances I got/get good effects from inhibiters, and two times out of two I felt worse from arginine (onliest precursor of NO).

I am avoiding arginine - apparently it encourages herpes family viruses, which I've struggled with. My doctor put me on citrulline and ornithine, both low on my NutrEval, and I started sleeping through the night!!

He has me testing NO after exertion and taking citrulline and ornithine then, too. I went canoeing last night then was low in NO, and took the citrulline and ornithine, some reduced glutathione, and some BCAAs, and no PEM, which I would have otherwise, this morning.

 

[Nine lives]

Have you measured it? My doctor is having me use Humann test strips, available on Amazon.

I am avoiding arginine - apparently it encourages herpes family viruses, which I've struggled with. My doctor put me on citrulline and ornithine, both low on my NutrEval, and I started sleeping through the night!!

He has me testing NO after exertion and taking citrulline and ornithine then, too. I went canoeing last night then was low in NO, and took the citrulline and ornithine, some reduced glutathione, and some BCAAs, and no PEM, which I would have otherwise, this morning.

Very promising! I bought the strips and for two mornings in a row I have undetectable levels. Have you settled on an optimal dose for citrulline and ornithine? I would like to give this a try.

 

[Learner1]

Very promising! I bought the strips and for two mornings in a row I have undetectable levels. Have you settled on an optimal dose for citrulline and ornithine? I would like to give this a try.

It seems to vary, depending on how low my NO is. I take them every day. Maybe 2-4g ornithine and 3-6g citrulline. The ornithine, when taken at night makes me sleepy and helps me sleep through the night. It does seem to make me sleepy during the day, though.