Preprint Oxidative Stress is a shared characteristic of ME/CFS and Long COVID, 2024, Shankar, Bonilla, Davis et al.

Did you notice that Ron Davis (Stanford) is one of the authors. If mistakes are made they should be corrected, let me clear about that.
But in my opinion Ron Davis deserves a lot more credit than the NIH, given his record on ME/CFS research. And his personal reason: finding a cure for his son, Whitney Dafoe. Dismissing the whole paper because of some misstakes?

 

#MECFS2023 Next Vishnu Shankar, immunologist in Mark Davis’ lab at Stanford is talking about oxidative stress and suppression of mitochondrial ATP levels in MECFS and Long Covid patients, and reactive oxygen species. He was inspired to work on ME by reading The Puzzle Solver book.

September 8, 2023
https://twitter.com/user/status/1700174741607432270

 

This seems to be in referrence to the "Methods Description" section in the supplementary material (can be downloaded here https://www.biorxiv.org/content/10.1101/2024.05.04.592477v1.supplementary-material), but unfortunately even that section doesn't contain the necessary details. It does state that 60% of LC patients meet IOM criteria for ME/CFS, but nothing about symptomology or severity of the acute infection of the LC group.

 

There was a guy in South Africa , Nash Petrovic, who supplied tailored antioxidant and other supps and had some successes among ME diagnosed. Of course the group is heterogenous (I believe anyway) but he may have helped some.

 

Of course, I’m very aware of that and I also noticed that one of the authors is Mark Davis who is a world-class immunologist for whom I have a lot of respect and similarly Bonilla is very well-known in the field and has a lot of expertise. All of these authors have my respect and obviously they know far more than I'll ever know about immunology, biology and genetics.

Others with far more expertise in the field have commented on the immunological aspects of the study already. I typically try to look at cohort characteristics, simply because I lack all the necessary background an immunologist has. But given the errors currently present, and even more importantly the lacking patient description, mentioned by @Hutan, which is strictly necessary to judge any LC data, there really is little for me to do and anything left to do seems like it isn't currently worth the energy for me, simply because if you perform a large array of tests on a very small sample set you can always find a positive result here and there simply by luck alone. I cannot tell if any of this data is causative and that's really the only thing that interests me. Should this work be followed-up by a larger study, I would be extremely happy to read it. Similarly if someone findings parts of the paper that are interesting and worth scrutinising I will pay attention to what they are saying.

One should not forget that it’s very possible to be dismissive of a paper, which I don't even think I've been, without being dismissive of the people having written it, any of their previous work or their intentions. To support this with an example: I have a lot of admiration and respect for the all of the work of the great Akiko Iwasaki, but that doesn't change the fact that the LC cortisol claims looked bogus to me.

I view the role of this forum to critically analyse data, independently of ones own beliefs or who the authors are.

 

Interesting, wondering if this was done as a trial i.e. biomarker (indicating oxidative stress), blinded - control group get placebo and trial group supplement?

 

My post above was based on working as a technician in a veterinary research laboratory which (among other things) looked at trace minerals/deficiency diseases. Selenium deficiency (which effectively equates to oxidative stress) was one of the two main areas (cobalt/B12 was the other). This was looking at the situation where selenium was severely limited - I guess you could have milder scenarios which are clinically relevant - perhaps sort of an echo of the problem i.e. rather than full blown selenium deficiency/oxidative stress as cause of pathology/disease.

 

As far as I know no. I am in the anything that helps camp, but I share any concerns about trumpeting sth as science which is not. about absolute and /or monolithic claims re treatments, aetiologes etc.

 

I get the impression the work on ME/CFS at Stanford is not particuarly well coordinated. I don't see research themes emerging. Are they trying to do to much at once?

The abstract for this paper does not give me faith, and people here have rightly pointed out several errors already.

 

"Using flow cytometry, we compared mitochondrial bioenergetic parameters in CD19 B cells as well as CD4 and CD8 T cell lymphocyte populations. Sample gating strategy for identifying these lymphocyte populations is shown in Supplementary Figure S1A. Although we did not identify any differences between healthy controls and ME/CFS or LC donors in mitochondrial mass or membrane potential (Supplementary Figure S2), decreases in mitochondrial ATP levels were observed across ME/CFS (1.6x decrease on average; HC v. ME/CFS two-sided t-test CD4 T cells p = 0.0458, CD8 T cells p = 0.0974, CD19 B cells p = 0.356) and LC donors (2.8x decrease on average; HC v. LC two-sided t-test CD4 T cells p = 0.0076, CD8 T cells p = 0.0182, CD19 B cells p = 0.0842), compared to healthy controls (Supplementary Figure S2). Our findings are consistent with a published study showing oxidative phosphorylation defects among PBMCs in CFS patients, compared to healthy controls69"

Unstimulated PBMCs are quiescent and when taken out of the body are also dying (every researcher around the world that I have spoken to who has measured this in time course experiments also finds that they die faster in ME/CFS). Naturally you will see generalised decreases in functional indicators and generalised increases in damaging or dysfunctional indicators in ex vivo disease cells that are falling apart at the seams. ROS could easily be a cause of this death/dysfunction but it could also be an consequence. This needs to be tested directly in a future study.

"Based on our results in Figure 3A, which also detected changes in acylcarnitine metabolism in ME/CFS donors, we compared lipid droplet levels using HCS LipidTox Green Neutral Lipid staining, which stains for neutral intracellular lipid droplets. As shown in Supplementary Figure 4G, we found lower lipid droplet levels in both LC and ME/CFS donors across all measured lymphocyte populations (ME/CFS: CD19 p = 5.19*10-4 , CD4 p = 4.58*10-4 , CD8 p = 3.01*10-4 ; LC: CD19 p = 0.140, CD4 p = 0.0669, CD8 p = 0.0759)."

This is a commercial dye so details about specificity and structure are scarce but one thing to bear in mind is that many neutral lipid stains are not specific to LDs and if you do whole-cell fluorescence measurements with them you will pick up signal from other cellular neutral lipids as well, not just LDs. Plasma membrane is like 40% neutral lipid or something significant like that from memory btw. This is why the chemists we worked with in the LD paper are making alternatives that are intended to be more specific. Non-LD background is a huge issue with a lot of neutral lipid dyes. I wish they did some LD imaging to complement this or that more information about the dye was available from Thermo.

Would have been good to see the kinetics of T cell stimulation between individuals compared given that the measurements were taken at the same 5 day time point. You can have cells from one individual start waning in response earlier than others particularly with this length of time from what I understand.

"We suggest that this lymphocyte dysfunction, driven by oxidative and mitochondrial damage, acts as an “energy sink”, much as an active infection does, draining the body of available energy and giving rise to debilitating fatigue and other sequelae."

Do people with ME/CFS gain less weight with comparable dietary intake? Do they have altered blood glucose? (genuinely asking, I don't know off the top of my head)

 

Thanks Hutan, that confirms my gut suspicions but I wasn’t certain enough to comment. Good to know.

 

@DMissa & @Hutan - Re the question on PwME gaining less weight with comparable dietary intake I can add that this is the case for my son & at least a couple of other teen & young adult males according to the mums in a FB group. I realise it is only anecdotal but I did calculate my son’s intake for several days & it was 3,700 - 4,000 to maintain his weight. Perhaps there is a subset of tall & very skinny males for who this is an issue, although they were much younger at onset and in my son’s case average height & weight for age at that time.

 

Yes, that's the case for my daughter too- so females also .
Re diabetes - given there seem to be issues with glycolysis, and the body work around may have different response re male/ female metabolism any poll would be interesting to see if there is a split manifest.

I think also that looking at liver function is a neglected area - both for detox, but also as the liver modulates many processes- many of which may play a part in ME/CFS - gluscose metabolism, hormone synthesis, immune response etc; the liver gut axis may be important given hints at microbiome dysbiosis too.

From a recent dip into diabetes for MIL, there seems to be a number of T2 diabetics who are insulin dependent ( i have heard 25% mooted , but don't know the source for this), so nothing is ever as clear cut as it may seem.