Study Shows that Red Blood Cells are Stiffer in People with ME

SWAlexander

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Intro:
Researchers have published a paper that shows various measures of deformability in the red blood cells of people with Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME/CFS) indicating that they are significantly stiffer in people with ME/CFS than those from healthy controls.

The observations suggest that red blood cell transport through microcapillaries may explain, at least in part, the ME/CFS phenotype, and promises to be a novel, first-pass diagnostic test.

Ronald W. Davis, PhD, who collaborated on the paper, writes: https://www.meaction.net/2019/01/02/study-shows-that-red-blood-cells-are-stiffer-in-people-with-me/
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SWAlexander

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Red Blood Cell Biomechanics in Chronic Fatigue Syndrome

DISCUSSION
Together, the various estimates show that the RBCs in CFS
patients are significantly less deformable than those of healthy controls.
We speculate that the larger and less deformable RBCs in CFS patients
may partly explain the musculoskeletal pain and fatigue in the
pathophysiology of CFS due to impaired microvascular perfusion and
tissue oxygenation. It has been shown that the quality of life of ME/CFS
patients was significantly worse as compared to patients with diseases
like sclerosis, cancer (multiple types, such as colon, breast and prostate),
type II diabetes, rheumatoid arthritis and chronic renal failure, among
others. This work introduces a new paradigm in our understanding of
the mechanistic aspects of ME/CFS. It also opens the possibility of a
diagnostic platform for ME/CFS using RBC deformability as the
biomarker.
Link to PDF: https://www.researchgate.net/public...Cell_Biomechanics_in_Chronic_Fatigue_Syndrome
 
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Of course, it is known that there is little NO production in ME/CFS patients. As reported in

Decreased NO production in endothelial cells exposed to plasma from ME/CFS patients
https://www.sciencedirect.com/science/article/pii/S1537189122000027

Most likely related to the GPCR autoantibodies spotted.


Without NO they will not be able to deform (https://www.ncbi.nlm.nih.gov/labs/pmc/articles/PMC6441278/). This is actually one of the challenges with stored donated blood, which leads to RBC rigidity.

But I have no idea why Ron still does not acknowledge that in the paper. It makes no sense to me talking about RBC rigidity in isolation.
 
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A standard way of increasing NO is with L-arginine, according to functional medicine, interestingly, citruline is high in watermelon, and the two metabolites, arginine and citruline, are intermediates in the urea cycle.

As for ferritin, I have low ferritin and found my energy is near pre-ME/CFS levels after supplementing with iron bisglycinate and my ferritin is going up, but I take a high dosage, 100mg of iron from iron bisglycinate. There are other forms of iron such as ferrous sulfate but the absorption is very low. I perform an experiment where I took nystatin, oral swish and swallow, 100,000 IU/ml and I felt back to normal and my ferritin went from 30 to 150 but after a few weeks it had no effect, my ferritin went back down to 30 and I was feeling back to my ME/CFS self. I am still trying other antibiotics for the candida infection that I still have and which my antibodies show I have a persistent infection. Also, note that I am in the process of curing my hashimotos, as evidence by high anti-TPO antibodies, with thymosin alpha-1, it is a peptide that I buy as a research chemical from canlab.net, other peptide companies charge too much and most of them are made in China where I don't trust the quality, in North America there are consequences for selling products that if they are found to be harmful, they could lose a lot.
 

SWAlexander

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A 2003 article:
New Markers of Inflammation and Endothelial Cell Activation
Current views regard atherosclerosis as a dynamic and progressive disease arising from the combination of endothelial dysfunction and inflammation.1–6 The vascular endothelium, located at the interface of blood and tissue, is able to sense changes in hemodynamic forces and bloodborne signals and react by synthesizing and releasing vasoactive substances. Vascular homeostasis is maintained by a balance between endothelium-derived relaxing and contracting factors. With disruption of this balance, mediated by inflammatory and traditional cardiovascular risk factors, the vasculature becomes susceptible to atheroma formation. Inflammatory mediators appear to play a fundamental role in the initiation, progression, and eventual rupture of atherosclerotic plaques. As evidence accumulates linking inflammatory processes to atherogenesis, markers of inflammation and endothelial activation may become useful by providing additional information about a patient’s risk of developing cardiovascular disease, as well as providing new targets for treatment.7,8 This review article is the first part of a two-article series examining emerging markers of inflammation and cardiovascular disease. Part 1 will provide a brief overview of the link between inflammation, endothelial dysfunction, and atherosclerosis and will begin highlighting emerging inflammatory mediators of endothelial cell (EC) activation, a discussion that will be continued in Part 2.
continue: https://www.ahajournals.org/doi/10.1161/01.cir.0000089190.95415.9f
 

halcyon

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Potentially another important note, also from your reference:
Simmonds et al. said:
Nevertheless, it is important to note that very high NO concentrations, as can be the case in various inflammatory states, may be deleterious for RBC deformability [49,56,84]. Thus, in summary, it appears that extracellular NO influences RBC structural and functional properties in a non-linear, dose-dependent manner
So it looks like it could possibly be either a too little or too much issue. My (probably inaccurate/incomplete) recollection is that NO activity tends to be increased in ME, and especially so after exertion, and several hypotheses involve this (Pall's NO/ONOO cycle, Maes & Morris' increased O&NS model, etc.).

A standard way of increasing NO is with L-arginine, according to functional medicine, interestingly, citruline is high in watermelon, and the two metabolites, arginine and citruline, are intermediates in the urea cycle.
It's possible that l-citrulline may be able to produce greater increases in plasma arginine than arginine itself can, due to metabolic effects (Schwedhelm et al., 2007). Also, combining l-citrulline with glutathione can produce a greater increase in NO than either alone (McKinley-Barnard et al., 2015).

That said, I'm not convinced that more or less NO will really help things, but NO synthesis and regulation is so damned complex that it's way too far beyond my ability to understand it.
 

mitoMAN

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Potentially another important note, also from your reference:

So it looks like it could possibly be either a too little or too much issue. My (probably inaccurate/incomplete) recollection is that NO activity tends to be increased in ME, and especially so after exertion, and several hypotheses involve this (Pall's NO/ONOO cycle, Maes & Morris' increased O&NS model, etc.).


It's possible that l-citrulline may be able to produce greater increases in plasma arginine than arginine itself can, due to metabolic effects (Schwedhelm et al., 2007). Also, combining l-citrulline with glutathione can produce a greater increase in NO than either alone (McKinley-Barnard et al., 2015).

That said, I'm not convinced that more or less NO will really help things, but NO synthesis and regulation is so damned complex that it's way too far beyond my ability to understand it.
This points towards a deficient NO production in ME/CFS

Decreased NO production in endothelial cells exposed to plasma from ME/CFS patients R Bertinat et al. Vascul Pharmacol. 2022.
https://pubmed.ncbi.nlm.nih.gov/35074481/
 

halcyon

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Isn't NO involved in the metabolic trap theory with the tryptophan buildup?

(tried to verify, didn't succeed)
It's been too long since I looked at it so I don't recall if they discuss that in the theory, but NO synthesized via iNOS is able to inhibit the IDO enzyme (Samelson-Jones & Yeh, 2006), so it's possible that NO could be involved.
 

halcyon

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This points towards a deficient NO production in ME/CFS

Decreased NO production in endothelial cells exposed to plasma from ME/CFS patients R Bertinat et al. Vascul Pharmacol. 2022.
https://pubmed.ncbi.nlm.nih.gov/35074481/
Yes, it does indicate specifically that something in ME plasma could be inhibiting constitutive synthesis of NO via the eNOS pathway in endothelial cells. But I believe that while that may be true, it does not provide any evidence that NO in general (especially induced via the other two pathways) is deficient in ME. There is evidence that points to the opposite. It's also possible that both of these things may be simultaneously true (diminished NO from endothelial eNOS, but elevated NO from iNOS).

Several studies have shown direct evidence of increased NO activity in ME, including elevated NO metabolites (Suárez et al., 2010), elevated iNOS production (Maes et al., 2007), and increased nitric oxide (Kurup & Kurup, 2003). Several more studies have shown indirect evidence of increased NO activity, including elevated levels of NOS by-product citrulline (Pall, 2002), elevated levels of methaemoglobin, which can be induced by nitric oxide (Richards et al., 2000), elevated levels of isoprostanes (Kennedy et al., 2005; Robinson et al., 2009), which can be induced by peroxynitrite (Shungu et al., 2014), and signs of elevated immune response to nitrosated compounds (Maes et al., 2006; Maes et al., 2008).
 
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apologies for being vague: my husband got test strips, to test saliva for NO (or some form of NO)...and he shows high levels of NO and I show nothing.

I'll try to come back and clarify this....where are the tests strips I don't know at the moment...
 

Learner1

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But I have no idea why Ron still does not acknowledge that in the paper. It makes no sense to me talking about RBC rigidity in isolation
I was sitting at the Stanford Symposium, thinking the exact same thing when this was presented, that the next step would be to look into oxidative and nitrosative stress and impact to membranes which would affect the rigidity of cell walls ... It's more than a mechanical problem - it's a biochemistry problem....
apologies for being vague: my husband got test strips, to test saliva for NO (or some form of NO)...and he shows high levels of NO and I show nothing.

I'll try to come back and clarify this....where are the tests strips I don't know at the moment...
Ive used Humann and Berkeley Life test strips, which showed I had low NO. I've been able to increase it using the Berkeley Life NO Booster, Kuvan, citrulline and/or ornithine. This NO increase has lowered by high blood pressure, by relaxing my blood vessels, as well as giving me more energy.
My (probably inaccurate/incomplete) recollection is that NO activity tends to be increased in ME, and especially so after exertion, and several hypotheses involve this (Pall's NO/ONOO cycle, Maes & Morris' increased O&NS model, etc.).
ME/CFS patients tended to be depleted in NO and have high oxidative and nitrosative stress forming peroxynitrites, which I think are using up the NO, leaving the deficiency. One would need to look up eNOS and iNOS to understand this better. Tetrahydrobiopterin (BH4) is also involved. Pall explains that C, folate, B12 and BH4 can reverse the vicious cycle of peroxynitrites. Low catecholamines are also a clue - I've found Kuvan (BH4)also increased my chronically low dopamine.
Fig-1-Tetrahydrobiopterin-BH4-is-an-essential-cofactor-all-three-hydroxylases-and.jpg

possible that l-citrulline may be able to produce greater increases in plasma arginine than arginine itself can, due to metabolic effects (Schwedhelm et al., 2007). Also, combining l-citrulline with glutathione can produce a greater increase in NO than either alone (McKinley-Barnard et al., 2015
Low BH4 can lead to low citrulline, arginine and ornithine, compromising the urea cycle and allowing ammonia to buildup, which can lead to unpleasant brain symptoms. Early on,noting my amino acid tests showed low citrulline and ornithine, and I tried taking them, and both solved my waking up at 2am problem, I think, due to reducing ammonia.

There's a whole system here, involving NO, OBOO and BH4 that affects blood vessels, catecholamines, urea cycle and more.
 

halcyon

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ME/CFS patients tended to be depleted in NO
Do you have any references that provide evidence of this? I spent several hours reviewing the literature for my post after the one you replied to, and I can find only a single study that measured NO directly in ME, while all the rest are looking at metabolites, synthase activity, or indirect measures, but all of which seem to support an increase in iNOS activity. I'm willing to believe NO itself could be depleted in ME, I just would like to see any direct evidence that might exist. It seems pretty obvious that iNOS should be upregulated in ME (and one study supports that), but I'd really like to see support for the notion that NO availability in vivo is then abnormal due to this.

I recall at least one study that found low citrulline in ME (which contradicts the evidence I posted above from Pall 2002), so would we explain that as evidence of either low iNOS activity, or instead as inadequate substrate (arginine/citrulline), and if the latter, why is there inadequate substrate (as you mentioned, can low BH4 explain it? is there any actual direct evidence for low BH4 in ME)?

It all feels roughly analogous to the tryptophan/IDO/kynurenine situation in ME, and is not really all that disconnected, as you point out with BH4 and catecholamines (indoleamines in this specific case) being involved here as well, and the fact that IDO and iNOS are driven by the same inflammatory signals (and as I mentioned elsewhere, IDO and iNOS have direct interactions too). There's conflicting evidence here as well, with some showing low tryptophan, and others high (which sparked the metabolic trap idea, as I understand it).

I've found Kuvan (BH4)also increased my chronically low dopamine.
Maybe you're not at liberty to discuss, but I'm curious how you're able to obtain it. After researching it for a while, I don't see any other realistic way to increase BH4 other than Kuvan, but my understanding was that it's only available with prescription for a very specific indication now.
 
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halcyon

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Ive used Humann and Berkeley Life test strips, which showed I had low NO. I've been able to increase it using the Berkeley Life NO Booster, Kuvan, citrulline and/or ornithine.
Are you not worried about doing further damage by increasing toxic NO metabolites by doing this, or do you feel you can offset that with other things (the supplements Pall advocates for example, as you mentioned)?

Again, it feels very similar to the tryptophan/IDO thing. Personally I disagree with the metabolic trap idea of tryptophan being elevated in ME, I think it's actually diminished and should perhaps be supplemented to try to increase availability for indoleamine synthesis (which I think is low in ME, and I think the evidence supports this), but at the same time doing so also provides more raw material for kynurenine synthesis and so could increase the amount of neurotoxic kynurenine metabolites if a person didn't do anything else to prevent that (and I still cannot fathom how OMF arrived at the idea of giving people l-kynurenine as a treatment).
 

mitoMAN

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I would also be interested in which dosage of BH4 Kuvan has helped you @Learner1

As your graphic shows BH4 depletion might be involved. However was there ever any study measuring BH4 in ME/CFS?
"eNOS activity is also dependent on cofactors and, in particular, 5,6,7,8-tetrahydrobiopterin (BH4), which is required to maintain the NO-generating enzyme dimeric state. "

I'm especiall interested in CNSA-001 (Sepiapterin) as it is acheaper alternative with higher efficency then direct BH4 supplementation.


Sepiapterin Administration Raises Tissue BH4 Levels More Efficiently Than BH4 Supplement in Normal Mice
https://link.springer.com/chapter/10.1007/978-1-4615-0945-5_33

while it was tested in healthy volunteers in gigantic dosages already.
Exploratory study of the effect of one week of orally administered CNSA-001 (sepiapterin) on CNS levels of tetrahydrobiopterin, dihydrobiopterin and monoamine neurotransmitter metabolites in healthy volunteers
https://pubmed.ncbi.nlm.nih.gov/31453106/
-> CNSA-001 is a pharmaceutical preparation of sepiapterin, a natural precursor of BH4 that induced larger increases in plasma BH4 compared with administration of the same doses of BH4 itself in healthy volunteers in a randomized trial.
 

mitoMAN

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Yes, it does indicate specifically that something in ME plasma could be inhibiting constitutive synthesis of NO via the eNOS pathway in endothelial cells. But I believe that while that may be true, it does not provide any evidence that NO in general (especially induced via the other two pathways) is deficient in ME. There is evidence that points to the opposite. It's also possible that both of these things may be simultaneously true (diminished NO from endothelial eNOS, but elevated NO from iNOS).

Several studies have shown direct evidence of increased NO activity in ME, including elevated NO metabolites (Suárez et al., 2010), elevated iNOS production (Maes et al., 2007), and increased nitric oxide (Kurup & Kurup, 2003). Several more studies have shown indirect evidence of increased NO activity, including elevated levels of NOS by-product citrulline (Pall, 2002), elevated levels of methaemoglobin, which can be induced by nitric oxide (Richards et al., 2000), elevated levels of isoprostanes (Kennedy et al., 2005; Robinson et al., 2009), which can be induced by peroxynitrite (Shungu et al., 2014), and signs of elevated immune response to nitrosated compounds (Maes et al., 2006; Maes et al., 2008).
This is a very impressive collection of studies you are linking. Indeed I am wondering now if NO is elevated or diminished. Can this be measured in serum by regular labs? Would be the easiest way to find out personally.

I had nitrotyrosine measured in 2020 and it was borderline high indicating elevated NO (probably due to iNOS) and peroxynitrite

So we would want to focus on elevating eNOS which seems dysfunctional.
 
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halcyon

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This is a very impressive collection of studies you are linking. Indeed I am wondering now if NO is elevated or diminished. Can this be measured in serum by regular labs? Would be the easiest way to find out personally.
Overall the evidence is a bit weaker than I had remembered, maybe there are a bunch of studies I missed or something. I'm left wondering the same as you.

I did find another study that seems to contradict the Maes et al. study. Ogawa et al. did not find an increase in iNOS expression in PBMCs from CFS patients relative to controls.