The RBC-NOX hypothesis

[anni66]

Cell d

"
If you look at a textbook of physiology, you’ll notice that oxygen transfer into the cell is primarily determined by ADP levels. If they’re low, there will be no oxygen transfer. That’s an important idea.

Of course, when you make ADP, you also make superoxide. And that’s a conundrum. An energy conundrum.
You cannot make energy without making oxidative stress in the form of superoxide.

The body knows this, of course, and it’s developed a fantastic redox cooling system composed of several different enzymes – SOD, GPx and catalase.

These are the kinetic speeds of these enzymes. These are the fastest enzymes systems in the human body. They’re incredibly speedy. And they have to be that way, because if they fail to take superoxide down to water, which is their job, then you cannot make energy. Because if you do, you’re just going to fry the mitochondrial membrane.

Because if this superoxide is not taken first to hydrogen peroxide and then to water by two different pathways, then the superoxide will turn into free radicals.

It will react with nitric oxide to form peroxynitrate. This is the OH/ONOO hypothesis of Marty Pall, which some of you may have heard or read about.

Or it reacts with hydrogen peroxide to form hydroxyradicals.

This impressive production of these free radicals will actually destroy the membranes – the mitochondrial membranes – and bring energy production to a halt.

The reason you do this is to save yourself. Because if you continue to generate energy and you cannot cool the system, then you have to bring down energy to save your life. And we think this is exactly what is going on.
In other words, the energy downregulation is not the problem.

The energy downregulation is the solution to prevent a deeper problem."

Cell danger response

 

[necessary8]

While nothing he said is exactly false, energy production is not the main source of ROS in many cases. Perhaps it was milions of years ago, but since then biology evolved mechanisms to purposfully create ROS, such as the NOX enzymes.

But he brings up an interesting point, that perhaps the decreased energy production that some groups observed in leukocytes (the seahorse studies) could be compensative downregulation in response to the oxidative stress. It's a bit of a long shot, but maybe.

 

[frozenborderline]

While nothing he said is exactly false, energy production is not the main source of ROS in many cases. Perhaps it was milions of years ago, but since then biology evolved mechanisms to purposfully create ROS, such as the NOX enzymes.

But he brings up an interesting point, that perhaps the decreased energy production that some groups observed in leukocytes (the seahorse studies) could be compensative downregulation in response to the oxidative stress. It's a bit of a long shot, but maybe.

Even if energy production was not the top cause of oxidative stress, if there were a ton of oxidative atress from another source wouldnt it make sense to downregulate energy production to compensate, even if responsible for only a portion of the oxidative stress? Anecdotally a few people with adhd before having me/cfs have noticed a huge change in tolerance of stimulants. I chalked this up to a possible change in threshold/tolerance for oxidative stress. Perhaps stimulants always produce oxidative stress but there's a certain amount the body can deal with normally. And me/cfs involves sources of oxidative stress that tip the threshold

 

[necessary8]

Even if energy production was not the top cause of oxidative stress, if there were a ton of oxidative atress from another source wouldnt it make sense to downregulate energy production to compensate, even if responsible for only a portion of the oxidative stress?

In pure principle, yes, it does make perfect sense. But I personally don't know quite enough about the specific pathways used to do this specific regulation, to say with full confidence that it definitely would happen. But I think it is an interesting idea, and I will keep it in mind.

 

[frozenborderline]

@necessary8 , I don't know enough about the specific pathways either. But besides that theory fitting with a lot of the anecdotal/observational data, i proposed it because two prominent me/cfs Researchers proposed it --paul Cheney and naviaux. When naviaux published his metabolomics paper I read it and saw that worms go into dauer because of lack of available food sources , needing to downregulate energy production for that reason. Well that obviously isn't tbe case for the vast majority of pwME so oxidative stress from some source (either infection or toxin or the immune system ) makes more sense as a reason for the body to downregulate metabolism. It seems that the vast majority of attempts to directly upregulate metabolism--through thyroid hormone, or other things that upregulate PDH, fail, or work a little bit temporarily, and this is why I agreed w naviauxs idea that the downregulation must be compensatory.

 

[frozenborderline]

Im reading this article https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4310837/ on sources of free radicals/ROS and it seems to back up Cheneys theory to an extent.

It also seems like the exogenous sources of free radicals mentioned, like pollution, radiation , etc , would dovetail with the proposed theory that mold only becomes particularly problematic by aggregating nanoparticles from industrial pollution (speculation that was validated by a recent pnas article). And of course the endogenous sources of free radical production include use by the immune system to kill pathogens.

What's interesting is that in ME/CFS we have an immune signature and overall metabolomic signature that indicates an infection but little evidence of this active infection. Its been suggested that this is because of a non cytolytic infection or latent virus. But I think it's possible that this is because the immune system is reacting to something that fits the signature of a virus but isn't a virus. Like nanoparticles. If a nanoparticle gets into the body it seems like the immune system will think there's a virus ans bombard it with free radicals, but macrophages are unable to clear it (I'm going to find the references for these claims in a second--this is not just speculation but is indeed backed up by research).

The first response to this theory would often be --why nanoparticles, there isnt any particular evidence for them being involved. Well, we know that industrial pollution creates health problems because It creates so many nanoparticles or ultrafine particles that get past the first line defense systems of the body ,able to pass through the nose and lungs without being expelled, and also able to get though innate immune responses. We also know tbat people who do "mold avoidance" tend to be avoiding not all molds but rather certain areas that are perceived to be bad which correlate more with industrial products and agriculture, moreso than with moisture vs dryness (people do well in rainforest and temperate rainforests areas , and poorly in some very dry cities in the southwest , such as Tucson).

This article, which I posted a relevant image from, shows that metal oxide nanoparticles can induce oxidative stress, cytokine production , and dna damage. https://www.tandfonline.com/doi/full/10.1080/21691401.2018.1468767
(@Hip this may interest you)

 

[SlamDancin]

@debored13 Haidut’s Ethyl Pyruvate, and seemingly B1 and Calcium, make me sleep, even during the day. I’ve learned not to step on PDH too hard. You’re perceptive on that one good call. I like a lot of what you’re saying in this thread. Is it possible to have mold levels checked ? Maybe I should just move because I know where I’m at must be moldy.

 

[frozenborderline]

@debored13 Haidut’s Ethyl Pyruvate, and seemingly B1 and Calcium, make me sleep, even during the day. I’ve learned not to step on PDH too hard. You’re perceptive on that one good call. I like a lot of what you’re saying in this thread. Is it possible to have mold levels checked ? Maybe I should just move because I know where I’m at must be moldy.

I'm going to respond to this in DM bc I want to be mindful of this thread getting a little off topic. Although my posts above were meant to be on how nanoparticles could be a relevant source of oxidative stress ill admit its all getting a little speculative and going off topic which is my bad

 

[Jackb23]

I am a fan of this hypothesis, I think that it can explain a lot of my symptomatology and also the conditions under which they worsen. I do not want to crash this thread by mixing it in with another but do want to allude to the this study: Identification of actin network proteins, talin-1 and filamin-A, in circulating extracellular vesicles as blood biomarkers for human myalgic encephalomyelitis/chronic fatigue syndrome. I think that exosomes and oxidative stress are both central players to this hypothesis.

In my situation I got profoundly better through deep breathing exercises (2 breaths a minute respiration rate or less for eventually 2 hours a day) and a large dose of bupropion (450 mg) in the fall of 2016. In fact, I got back to around 90%, but when I try to convey that percentage to those that aren't in the ME/CFS community, I am not quite sure they completely understand the suffering that can still entail. To me, even those affected the most mildly with ME/CFS can have a severe rate-limiting step which is this illness. To me, when I am describing functional percentages it's more akin to describing how big of a hole there is in a canoe when it is put in water.

Despite regaining a large part of my functioning, in the winter of 2017 I became ill with pneumonia and I experienced a profound relapse/setback. Now, the Trap Hypothesis could potentially explain my setback in a sort of "re-trap" (I guess), but I didn't know how it might apply to this hypothesis. After mulling it over this is what I think may explain it:

When I did my deep-breathing, I believe that I got somewhat upstream towards the root of the illness, but not quite all of the way there. Vagus nerve stimulation has been shown to decrease oxidative stress and inflammation in the brain (See below paragraph). The large breaths of air would offer even further benefits and it is hard to parse out which one was really helping, if not both.

1. Tsutsumi T., Ide T., Yamato M., Kudou W., Andou M., Hirooka Y., Utsumi H., Tsutsui H., Sunagawa K. Modulation of the myocardial redox state by vagal nerve stimulation after experimental myocardial infarction. Cardiovasc. Res. 2008;77:713–721. doi: 10.1093/cvr/cvm092.

2. M. Chen, X. Zhou, L. Yu et al., “Low-level vagus nerve stimulation attenuates myocardial ischemic reperfusion injury by antioxidative stress and antiapoptosis reactions in canines,” Journal of Cardiovascular Electrophysiology, vol. 27, no. 2, pp. 224–231, 2016.


Would a large suppression of symptoms or a remission better explain my improvement and then set-back?- I have come to believe it is the former. Pneumonia can have its own profound effects, but it wasn't the cause of my illness in the first place, Mono was.


In this study, Mycobacterium tuberculosis reactivates HIV via exosomes mediated resetting of cellular redox potential and bioenergetics (https://www.biorxiv.org/content/10.1101/629048v1), the authors found that Mycobacterium tuberculosis could reactivate HIV exosomes through oxidative stress. Epstein-Barr and Streptococcus pneumoniae interactions would need to be studied specifically, but I do believe the pneumonia could have reactivated the Epstein-Barr exosomes through extreme oxidative stress in a similar mechanism. One is even a bacteria and the other a virus like the study above.

Epstein Barr has also been found to play a hand in Lupus, and perhaps through exosomal inflammation specifically: "Recently, we discovered that EBV-infected B cells secrete highly inflammatory viral RNAs (EBERs) via extracellular vesicles (EVs)."
Inflammatory Exosomes Implicate a Role for Epstein Barr Virus Infection in the Pathophysiology of Systemic Lupus Erythematosus, 2015

It has also been found to play a role in Epstein-Barr associated cancers: "EBV-associated exosomes undoubtedly play pivotal roles in the pathogenesis of the viral infections and EBV-associated cancers especially in NPC. The cumulative findings suggest that the EBV-associated exosomes are ideal targets for the development of diagnostic biomarker and anticancer therapy."
Pathogenic Role of Exosomes in Epstein-Barr Virus (EBV)-Associated Cancers, 2017



More on EBV and exosomes:

"
EBV mRNA(+) Exosomes
Besides viral small RNA and miRNAs, a study has recently shown that EBV protein-coding RNAs also exist in the virally modified exosomes [73]. Four key latent gene transcripts were detected, including LMP1, LMP2, EBNA1, and EBNA2. However, it is still unclear if these viral mRNA can be delivered into the recipient cells and if these transcripts are biologically active in the uninfected recipient cells."
Extracellular Vesicles in Epstein-Barr Virus’ Life Cycle and Pathogenesis, 2019


Whether or not exosomes cause the oxidative stress or vice versa, I have no idea-- there may even be a third player involved that is generating both from upstream. I do think that your hypothesis, @necessary8 , can explain a large part of what has been going on in my body specifically, however. And I appreciate all the time and thinking that you gave this specific hypotheses.

 

[SlamDancin]

@Jackb23 Hey great post and research. I would suggest that you are missing out on hypoxia and Dr Van Elazzaker work is showing what I think is a complete breakdown of neurovascular coupling in this illness. Thats the CNS telling the peripheral vasculature how much blood to supply the brain. So basically when we exercise we put ourself into brain hypoxia and one patient had almost no function at al. Scary. He’s using impressive diagnostic criteria and new technologies to show that I guess no matter the setting off point for pwme there are unanimous changes in NVC under exercise challenge. I believe exosomes are involved in the bodies’ response to hypoxia but I haven’t studied it too much. NMDA, Nitric Oxide and GABA receptor containing Somatostatin receptors are involved and it’s a topic I’m hoping to talk with a real researcher about. Somatostatin is an understudied culprit I believe as it stops the motor activity of rats, something that struck me as sounding a lot like extreme fatigue. Somatostatin receptors can be activated in hypoxia I believe if NVC fails. It’s possible that if you have reactivated EBV it attacked the brain stem mostly because for some reason I think it likes the spine to set up shop. It’s nasty and I do believe it can “cause” or jump off a number of chronic illnesses when reactivated.

 

[Jackb23]

@SlamDancin I have been keeping up with Dr Van Elazzaker and his preliminary findings regarding the hypoxia and the neuroinflammation. I think it all ties in well with this hypothesis if you are to think about the effects reactive-oxygen-species have on erythrocytes. I’m not quite sure Van Elazzaker is to the point where he can use his findings as a diagnostic, but I do think he is finding some very interesting things

 

[sb4]

@Jackb23 I am interested in the breathing exercises that helped. How did you get to 2 breathes a minute? How long breathing in, breathing out? Was it Buteyko or something else?

 

[Jackb23]

@sb4

I started out by counting during my exhales, but then slowly moved to counting at the end of my exhale. It’s hit or miss in that if I am not able to relax and really focus solely on my breath I will average around 4 breaths a minute. It’s extremely hard for me to average two breaths a minute for much longer than 10 minutes. What I’ll do is I’ll set either a timer or a stop watch and either set it to 10 minutes or to see where I’m at time wise after 20-25 breaths.

When I first started, my respiration rate was not that low, but with time I slowly got it better/slower. I have come to realize that the rate is highly contingent upon my ability to not be scattered. Also, a good way to ensure that you’re getting a good deep breath is to breath out first instead of in. Expelling all the air you have first will allow you to take in much more air. Also, the slow respiration rate is something I kind of have to settle in to. Almost like how people step into a cold pool.

At my best, my inhale amounts to around 6-7 seconds—exhale to around 20+. I find that when I focus on my inhale it makes it much, much harder to relax so I don’t really do that at all. I solely focus on the exhale portion.

I can also tell when I am doing well and slowly down my respiration remarkably as my veins pop in places they normally don’t and quite heavily. This vasodilation may play a huge part in the benefits

I am not familiar with the buteyko method, but after googling and looking it over I could see it being of benefit.

 

[sb4]

@Jackb23 I just gave it a go. Definitely feel warmer however my heart pounding noticeably picks up on long exhale. Perhaps the vasodialation is contraindicated in some POTS cases.

Around 2 years ago I noticed when my POTS was bad I would feel out of breath and be gasping for air. I stopped this and it seemed to help a little.

 

[necessary8]

Okay, so, before we get ahead of ourselves, I will clarify.
I am aware of dr Eguchi's paper. I've talked to him directly about it. It nullifies half of this hypothesis - he found no NOX in the exosomes from ME/CFS plasma, and his methodology looks solid. So everything here that hinged on there being NOX exosomes goes out of the window.
On the other hand, the other half of this hypothesis, that there is abnormally high extracellular oxidative damage in ME/CFS, which might be what's causing the RBC deformability issues, which might explain symptoms - this part I believe is still very solid. I just need to figure out what's causing it. At this moment, I don't know. Once I have a good idea, I will probably update but it might take a long while.

 

[bread.]

got it by now?

 

[necessary8]

Yeah, but I'm not the one who found it, dr Prusty is. The suppression of SOD2 due to mitochondrial fission can explain the oxidative damage. We joined our hypotheses.

 

[wigglethemouse]

We joined our hypotheses.

Does that mean you are in contact with Dr. Prusty @necessary8 ?

 

[necessary8]

Does that mean you are in contact with Dr. Prusty @necessary8 ?

Yes. We talk via email and have a very loose and informal collaboration. I'm doing some bioinformatics for him right now, we'll see if it goes anywhere, not sure at the moment yet.

 

[frozenborderline]

https://www.nature.com/articles/ncomms7245
A nature article on purinergic signalling and immune cells, and infection

https://www.nature.com/articles/ncomms7245

Hope this can be helpful