OMF-funded RBC Deformability Paper Published!

[wigglethemouse]

Full text of the paper has been published :
Red blood cell deformability is diminished in patients with Chronic Fatigue Syndrome, Davis et al

Abstract:

BACKGROUND: Myalgic encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a poorly understood disease. Amongst others symptoms, the disease is associated with profound fatigue, cognitive dysfunction, sleep abnormalities, and other symptoms that are made worse by physical or mental exertion. While the etiology of the disease is still debated, evidence suggests oxidative damage to immune and hematological systems as one of the pathophysiological mechanisms of the disease. Since red blood cells (RBCs) are well-known scavengers of oxidative stress, and are critical in microvascular perfusion and tissue oxygenation, we hypothesized that RBC deformability is adversely affected in ME/CFS.

METHODS: We used a custom microfluidic platform and high-speed microscopy to assess the difference in deformability of RBCs obtained from ME/CFS patients and age-matched healthy controls.

RESULTS AND CONCLUSION: We observed from various measures of deformability that the RBCs isolated from ME/CFS patients were significantly stiffer than those from healthy controls. Our observations suggest that RBC transport through microcapillaries may explain, at least in part, the ME/CFS phenotype, and promises to be a novel first-pass diagnostic test.

Here is the meat and potatoes of the paper

By comparing the change in cell diameters normalized to the width of the test channel before (L1/d) and after (L2/d) entry, Fig. 1D clearly demonstrates that RBCs from ME/CFS patients deform 7-fold less compared to those from the HC.

 

[Belbyr]

Fantastic! On to the next step!

 

[halcyon]

Full text of the paper has been published :
Red blood cell deformability is diminished in patients with Chronic Fatigue Syndrome, Davis et al

Zero reference to, or even acknowledgement of, the prior published research of Les Simpson that this study reproduces the finding of, or that it was his original idea to use this as a diagnostic test? Bizarre.

 

[Janet Dafoe]

Zero reference to, or even acknowledgement of, the prior published research of Les Simpson that this study reproduces the finding of, or that it was his original idea to use this as a diagnostic test? Bizarre.

That was supposed to be in there. Ron will figure out what happened.

 

[kangaSue]

result in the body producing less heme.

A deficiency in heme oxygenase (HO-1) is one of the more significant clinical findings in those with idiopathic gastroparesis too.
HO-1 is known to have major immunomodulatory and anti-inflammatory properties ad I'm not sure on this but I think I have seen somewhere that an HO-1 deficiency can be the case sometimes in IBS too.

 

[keepontruckin]

More than fantastic. It's heaven sent.

 

[Research 1st]

That was supposed to be in there. Ron will figure out what happened.

Hi Janet, Happy New Year and to all reading this. Let's hope we make some further ground this year.

I had Les Simpson's RBC test many years ago, maybe around 20. Back then we pricked our finger and put the blood between a glass slide which had a chemical we added to immediately use to fix it. Formaldehyde maybe.

What's interesting is Ron's research shows a finding that on average ME CFS patients RBCs were 7 fold less elastic which is a huge change.

This interests me as my old Les Simpson result showed under a microscope (he provided an image) my RBC's were around 80%+ deformed in shape, so a morphological change that is also high like Ron's.

Back then Les Simpson said he thought this could be from oxidative stress, infection and also that this would impair blood flow causing pain and if I recall correctly would affect oxygenation and maybe nutrition also.

A few quotes from Les are available here in summary format that may be of interest.

http://www.cfidsreport.com/Articles/researchers/lessimpson.htm

Thanks.

 

[wigglethemouse]

A few quotes from Les are available here in summary format that may be of interest.

http://www.cfidsreport.com/Articles/researchers/lessimpson.htm

I really liked two of the quotes.

Thanks to OMF funding Ron & wider team have the ability to facilitate Les Simpsons vision if they can build the microfluidic tester and validate findings in larger cohorts.

An attempt by Simpson to establish a clinic to test red blood cells did not survive due to lack of funds, a problem experienced by many researchers interested in CFIDS. If Simpson’s ideas are correct, agents which improve blood viscosity/flow may relieve symptoms in CFIDS patients. He hopes to someday see placebo-controlled studies conducted “with the primary objective of determining which agent [haemorheological], if any, has the ability to improve patient well-being.” He continues, “Because it is not possible to increase the diameter of a capillary, treatments should be aimed at increasing red cell flexibility.

This one really hits home. It astonishes me that Les Simpsons work has been out there for so many years, but "not invented here" syndrome has affected so many researchers that his work has not been built upon until now. By researchers who believe working collaboratively is key.

He is deeply concerned that closed minds and personal agendas are keeping knowledge of the illness from progressing,-- as if Kuhn's ideas are being played out in a real-time drama. Researchers, he feels, seem more interested in promoting their own fields of expertise and presumptions, rather than working collaboratively to defeat a disabling illness.

 

[Kenny Banya]

...Based on extensive literature search, we found only one of the drugs, colchicine, which was prescribed to one patient, has been shown to slightly affect cellular deformability. However, the deformability parameters for this particular patient were well within the range for other patients...

This would suggest that Colchicine makes little difference to red blood cell deformability in MECFS patients

 

[Mel9]

More than fantastic. It's heaven sent.

Yes, marvellous!

@Rose49, does Ron and his team have plans to investigate the effect of Trental on RBC deformability?

 

[Belbyr]

I looked at the picture posted on OMF here, it reminded me to pull out my Fry Labs (controversial lab) and compare from about 10 years ago. https://www.omf.ngo/2018/04/04/rbc-shape-rbc-deformability/

First picture is when I started lyme treatment, second picture was about 4 months in to IV treatment... No one could really say why my RBC's looked like that...

 

[Murph]

Weak RBC deformability has the power to explain a lot of our symptoms even if it is not the ultimate upstream cause of the disease.

Deformability is about more than just what tubes blood cells can fit through.

VASODILATION

The stretchiness of your red blood cells is important not only to determine which channels the cells can go down but to make the blood vessels themselves relax. Deformable red blood cells release ATP (the energy molecule) which causes vasodilation, as explained in this paper:

The dual roles of red blood cells in tissue oxygen delivery: oxygen carriers and regulators of local blood flow

Frank B. Jensen
Journal of Experimental Biology 2009 212: 3387-3393; doi: 10.1242/jeb.023697

http://jeb.biologists.org/content/212/21/3387

Mammalian RBCs release ATP when exposed to reduced oxygen tensions, and the amount released is linked to the decrease in Hb O2 saturation (Ellsworth et al., 1995; Jagger et al., 2001). The extracellular ATP diffuses to the endothelium, where it binds to P2y purinergic receptors, which activates the synthesis of vasodilators (including nitric oxide) that relax vascular smooth muscles and increase local blood flow and O2 delivery (Ellsworth et al., 1995; Sprague et al., 2007).
...
ATP release is activated not only by PO2 decrease but also by mechanical deformation (Sprague et al., 2001), as would occur when RBCs are squeezed through narrow vessels. The signal transduction seems to involve activation of G protein and adenylyl cyclase with the accumulation of cAMP (Sprague et al., 2001; Sprague et al., 2007). The exact membrane conduit for ATP release remains to be identified (Sprague et al., 2007), but, as mentioned above, a recent study points to the gap junction protein pannexin 1 that is expressed in human RBCs (even though they do not form gap junctions) and forms an ATP-permeable channel in the plasma membrane (Locovei et al., 2006).

As I understand it, the physical bumping around that happens when a bunch of red blood cells arrive at a narrow blood vessel causes them to spurt out ATP, which make the endothelium relax and the blood vessel open up. If you have low blood volume presumably there is less bumping of cells and less vasodilation..

PERSONALLY

What is interesting to me is that exercise and alcohol - both of which cause vasodilation - are dangerous to me. Both cause PEM. I suspect a failure of my body to do vasodilation properly is the cause of the PEM.

HYDRATION

A second interesting point: hydration affects RBC deformability as this picture below shows.

Like a lot of people here, I am always thirsty. One theory is I'm constantly trying to climb back off the furthest part of this curve. I doubt that though. I think I'm on the left side of the peak. It is more likely that by compensating for low blood volume with lots of water I'm actually diluting my blood and I'm on the first part of the curve, reducing my RBC deformability. I think this theory is more likely because I am constantly hungering for salts and electrolytes i.e. trying to increase osmolarity....

CHOCOLATE

I've started eating a lot of raw cacao powder (high in a polyphenol called epicatechin) and a lot of cocoa (high in a polyphenol called catechin (so long as it has not been dutch processed. Non ducthed cocoa is hard to find outside the USA!)). Research suggests these compounds are good for RBC deformability:

https://www.ncbi.nlm.nih.gov/pubmed/28314639

Results so far are neither disappointing nor dramatically good.

 

[Murph]

There exists a disease that makes your red blood cells extremely sensitive to oxidative stress. That disease is called G6PD deficiency. It causes huge decreases in red blood cell deformability when cells are hit with oxidative stress as this next graph shows.

Source: Inability to maintain GSH pool in G6PD-deficient red cells causes futile AMPK activation and irreversible metabolic disturbance.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4361223/

What's interesting about it is that it also causes big metabolic defects in the cells.

Here's how a bunch of metabolites looked after an oxidative stress challenge:

G6PD deficiency is a genetic disorder and it is quite common in Asia so this is not what we've got. The point of this comment is that there is precedent for a mega-response to oxidative stress and that oxidative stress can include a big set of reactions in red blood cells including metabolic ones.

 

[wigglethemouse]

PERSONALLY

What is interesting to me is that exercise and alcohol - both of which cause vasodilation - are dangerous to me. Both cause PEM. I suspect a failure of my body to do vasodilation properly is the cause of the PEM.

The more I "Google" erythrocyte the more interesting it gets. Searching "erythrocyte alcohol" brings up lots of interesting papers e.g.
The Effect of Alcohols on Red Blood Cell Mechanical Properties and Membrane Fluidity Depends on Their Molecular Size

These results are in accordance with available data obtained on model membranes. They document the presence of mechanical links between RBC deformability and near-surface membrane fluidity, chain length-dependence of the ability of alcohols to alter RBC mechanical behavior, and the biphasic response of RBC deformability and near-surface membrane fluidity to increasing alcohol concentrations.

Prevention of ethanol-induced erythrocyte transformations by fructose and natural honey in low alcohol tolerance mice

The effects of fructose and natural honey in preventing the ethanol-induced transformations of erythrocytes on DBA/2J mice were studied. Ethanol administration (6 g/kg body weight (b.w.), p.o.) caused three types of erythrocyte deformation under a light microscope, i.e., echinocytes at stages I and II, echinocytes at stage III, and stomatocytes. The natural honey group (2.5 g/kg b.w., p.o.) showed the lowest percentage of the deformed erythrocytes. The fructose group (2.0 g/kg b.w., p.o.) showed a lower preventive than the group (1.0 g fructose/kg b.w., p.o.), which suggests that the administration of a large amount of fructose would reduce the metabolic rate of ethanol. A remarkable increase in erythrocyte diameter was caused by the ethanol administration, while the increase was suppressed the most in the natural honey group.

Flavonoids may be an interesting avenue to pursue (they can also be helpful for MCAS), in addition to the Omega 3/6 and EPO that Les Simpson was interested in
Phenolics from monofloral honeys protect human erythrocyte membranes against oxidative damage.

Overall, this study indicates that honey contains relevant antioxidant compounds responsible, at least in part, for its biological activity and that uptake of its flavonoids may provide defense and promote cell functions in erythrocytes.

I thought this was an interesting snippet in this paper
Protective effect of flavonoids against red blood cell hemolysis by free radicals

RBCs are more frequently exposed to oxygen than other body tissue and, thus, are more susceptible to oxidative damage.

 

[Murph]

I looked at the picture posted on OMF here, it reminded me to pull out my Fry Labs (controversial lab) and compare from about 10 years ago. https://www.omf.ngo/2018/04/04/rbc-shape-rbc-deformability/

First picture is when I started lyme treatment, second picture was about 4 months in to IV treatment... No one could really say why my RBC's looked like that...

Looks like your RBCs were going through a punk phase

Did the IV treatment make you feel worse or better?

EDIT: I found this picture in a 2010 paper :

Effects of ATP on morphology and dynamic fluctuation in RBC membrane.
(A) Topography of a healthy RBC,
(B) of an ATP-depleted RBC (irreversible-ATP group),
(C,) of an ATP-depleted RBC (metabolic-ATP group),
and
(D) of a RBC with recovered ATP level (+ATP group),
(E–H) Instantaneous displacement maps of membrane fluctuation in the Fig. 1A–D, resp.
The scale bar is 2 μm. The colorbar scales are in μm and nm, resp.

Perhaps your spiky RBCs were low on ATP?

 

[Gemini]

Zero reference to, or even acknowledgement of, the prior published research of Les Simpson that this study reproduces the finding of, or that it was his original idea to use this as a diagnostic test?Bizzare

Yes, @halcyon Simpson's ME/CFS research, dating back to the 1980's, is listed on PubMed.

And for those not familiar with it a good starting point is Chapter 65 of the book "The Clinical and Scientific Basis of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome" which is dedicated to it.

[Book available online from the Nightingale Research Foundation and amazon.com]

@Janet Dafoe (Rose49)

 

[Gemini]

It astonishes me that Les Simpsons work has been out there for so many years, but "not invented here" syndrome has affected so many researchers that his work has not been built upon until now. By researchers who believe working collaboratively is key.

Yes, @wigglethemouse.

Twenty years ago on a trip to the US Simpson reached out to our patient organization to enlist volunteers for a new study, to a leading ME/CFS clinician to lead it, and to the Bioengineering Department of a major university to develop a "bioengineering model" of RBC abnormalities in ME/CFS. He also wanted to explore patient testing protocols and treatments.

It would be nice to acknowledge his research contributions to the field as well as his dedication to helping ME/CFS patients.

@Janet Dafoe (Rose49)

 

[Tally]

It astonishes me that Les Simpsons work has been out there for so many years, but "not invented here" syndrome has affected so many researchers that his work has not been built upon until now.

To be fair the whole ME field suffers horribly from lack of ANY reproduced research, and that's totally on people deciding what to fund, not on researchers.

This research was made possible by OMF's fundraising capability and was entirely funded by donors (many of whom are impoverished patients). I don't know how NIH and other neglectful governments around the world don't die of shame.

 

[Kenny Banya]

The lead author on the study indicated on Twitter that they will be doing further investigations that will include the correlation between disease severity & deformability

 

[Tally]

The lead author on the study indicated on Twitter that they will be doing further investigations that will include the correlation between disease severity & deformability

Here it is

https://twitter.com/i/web/status/1080316332493524997