The nanoneedle salt stress test – too good a clue to leave abandoned on the lab bench?

A few thoughts

I don't understand why tenure was quite so big an issue. Every academic wants tenure as it gives them security for the rest of their career. But I think it's unusual to get it very early on, and not getting it quickly doesn't cost you your job - you just don't have job security locked in. Rahim Esfandyar-Pour still has his job and his lab. But of course, it is entirely his choice what work to pursue, and he already did the ME community a favour with his early nanoneedle work.

I realise that Ron Davis has very high ethical standards, but as someone with an awful disease, I wish there had been a way to follow up on the initial findings (that no longer seems possible as RE is no longer responding to RD).

It is very good this is now being followed up in the UK. I hadn't realised that Prof Robert Dorey was a nanomaterial expert (though not in medical applications), which is great.

Sid said:
The blurb from ME Research UK implies that they were able to detect a difference between ME and MS, though I should add that a statistically significant difference between groups does not necessarily mean the test can differentiate in a clinically useful manner.
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Exactly. It was the spectacular differences between patients and controls, with clear blue water between them, that was so compelling. I appreciate @Jonathan Edwards's concerns, but I still want this finding followed up robustly.

And I am excited about this, not least the focus on understanding any pathology behind the differences. If those differences are indeed real. With such a big effect, you might expect the underlying biology to be a big clue to a cause of MEcfs.
 
Simon M said:
It was the spectacular differences between patients and controls, with clear blue water between them, that was so compelling.
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That's my big worry, that it was almost too good to be true. Can't think of other ME research that had such differences between patients and controls. My other big worry that this is the real reason progress hasn't been made, it was just an artifact of the testing and Ron has realised this and let it quietly fall away.
 
Has anyone not affiliated with RD tried getting in touch with RE to find out why this project was abandoned? I always assumed it didn’t pan out in subsequent testing so they just quietly dropped it (like a bunch of other things they used to talk about but don’t anymore). So, this news of an independent replication in the UK seemingly out of nowhere is very surprising.
 
Sid said:
Has anyone not affiliated with RD tried getting in touch with RE to find out why this project was abandoned? I always assumed it didn’t pan out in subsequent testing so they just quietly dropped it (like a bunch of other things they used to talk about but don’t anymore). So, this news of an independent replication in the UK seemingly out of nowhere is very surprising.
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Surely the charities must have known more before deciding to allocate funding? But perhaps not?
 
Binkie4 said:
Surely the charities must have known more before deciding to allocate funding? But perhaps not?
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I am not sure that in this situation it is ever possible to 'know' much more. The results of the original study are out there. No information is available as to exactly why it was not followed up and nobody can be forced to provide that.

The charities use people like me to judge whether or not repeating such studies - or doing similar experiments that might corroborate the findings - is worth investing in. I agree with Simon that whatever the uncertainties, an attempt to confirm has to be worth at least a small investment. One has to trust that researchers will do their best to get a genuinely meaningful result.
 
John Mac said:
My other big worry that this is the real reason progress hasn't been made, it was just an artifact of the testing and Ron has realised this and let it quietly fall away.
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I did wonder about that, but I don’t think you pursue an NIH grant for a dead duck. And it
does seems to have come down to the fact that RE no longer wanted to pursue the work, and he was the expert,

as for “too good to be true “– I completely agree. All the more reason for replication,
 
Sid said:
Has anyone not affiliated with RD tried getting in touch with RE to find out why this project was abandoned? I always assumed it didn’t pan out in subsequent testing so they just quietly dropped it (like a bunch of other things they used to talk about but don’t anymore). So, this news of an independent replication in the UK seemingly out of nowhere is very surprising.
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You've expressed my thoughts perfectly there Sid.
 
Here is some background on Professor Michael Hughes, one of the P.I.'s at Surrey University on the ME Research UK and ME Association funded study to develop a diagnostic biomarker.
Source - https://www.surrey.ac.uk/people/michael-hughes

In short they developed a technique to measure the electrical properties of cells quickly and can differentiate the cells by their electrical properties.
Research interests
My research is in the area of bioelectronics, most specifically in a phenomenon of dielectrophoresis (DEP). DEP is the name given to micro- or nano-particle movement due to the interaction with certain kind of electrical field; the speed and direction of movement depends on the resistance and capacitance of the particle and the frequency of the electric field; analysing the response as a function of frequency allows those properties to be measured, whilst mixtures of particles with different properties can be separated at frequencies where their responses are different. Whilst my group has looked at a range of particles including proteins, DNA, nanowires, carbon nanotubes and clay, the primary focus has been on cells - including cancer, stem cells, bacteria and yeast.
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They developed a disposable chip to do the high throughput measurements and it has been commercialized or is in the process of being. Company and product info can be found here - https://deparator.com/
My principal line of research is the development of new technology to exploit the DEP effect. We have developed a new chip technology - called the DEP-Well - which has dramatically increased the speed, accuracy and throughput of technology. For example, we have built a tool for measuring the mean electrical properties of 20,000 cells in 10 seconds (current gold-standard technology measures one cell in 5 hours); and have recently build the world's fastest cell separator, capable of sorting 200,000 cells per second. Both of these technologies require no special chemicals, needing only a low-cost disposable chip; both have also been commercialisd, with the former now being sold as the DEPtech 3DEP and the latter as the DEParator.
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I thought the mention of detecting cell death was interesting.
We have used the technology to meet a number of scientific needs, including an electrophysiology-based test for oral cancer and a new method for sorting blood cells. Investigations using the technology have given new insights into cell death and responses to anti-cancr treatment.
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Several researchers have reported that ME/CFS PMBC's die quicker than healthy cells. I think the Morten Group mentioned it in a report to the ME Association (perhaps Myhill replication work?) and @DMissa and colleagues reported it in their Lymphoblast mitochondrial analysis papers.
One of the theories discussed about the nanoneedle test was perhaps the salt stress was causing cells to die quicker.

Edit to add : Dr Hughes is now at Khalifa University in the United Arab Emirates and visiting professor at Surrey.
https://www.linkedin.com/in/michael-pycraft-hughes-3a77945/?originalSubdomain=ae
So who is leading the day to day work of the ME/CFS salt stress study as Dr Labeed is also in the UAE?
 
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There is a video and written description of the DEParator cell separator here
https://deparator.com/elementor-204/
  • The DEParator is a cell separation system that utilises the innate but differing properties of cells in suspension based on Dielectrophoresis (DEP)..
  • The DEParator draws the cell mixture through a patented DEPwell structure with over 600 active pores,
  • Each pore has a multi-layer structure that sets up a periodically repeating non-linear electric field across the flow path at a user defined frequency. As the cell mixture flows through the wells, some cells experience positive DEP and move into the fields to become trapped at the wall. Others experience negative DEP and are repelled into the centre of the well and remaining in the flow, hence are expelled.
  • The system can either trap wanted or unwanted cells, concentrating the desired cell type. Repeated flows at differing frequencies thus sequentially remove the unwanted cells, leaving only the desired ones, in a benign fashion WITHOUT any form of labelling unlike other cell separation systems such as magnetic beads or FACS
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I thought their papers on apoptosis could be interesting in relation to the salt stress test. Here is one of them where they use the detection method to look at apoptosis with relation to drug discovery in cancer.
Rapid assessment of early biophysical changes in K562 cells during apoptosis determined using dielectrophoresis
Sue Chin,1 Michael P Hughes,1 Helen M Coley,2 and Fatima H Labeed1
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2426800/

Apoptosis, or programmed cell death, is a vital cellular process responsible for causing cells to self-terminate at the end of their useful life. Abrogation of this process is commonly linked to cancer, and rapid detection of apoptosis in vitro is vital to the discovery of new anti-cancer drugs. In this paper, we describe the application of the electrical phenomenon dielectrophoresis for detecting apoptosis at very early stages after drug induction, on the basis of changes in electrophysiological properties. Our studies have revealed that K562 (human myelogenous leukemia) cells show a persistent elevation in the cytoplasmic conductivity occurring as early as 30 minutes following exposure to staurosporine. This method therefore allows a far more rapid detection method than existing biochemical marker methods.
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Here are all the papers listed on the DEParator company website.
https://deparator.com/elementor-321
 
The Surrey University page for Dr Labeed is here:
https://www.surrey.ac.uk/people/fatima-labeed

In the research interests section she mentions developing drug assays, being able to do rapid apoptosis measurement, and cytoplasmic resistance measurement.
Finally, a third line of research has been the development of drug assays using DEP to monitor electrophysiological changes more rapidly or more accurately by other means. These include very rapid apoptosis measurement, effect of ion channel blockers on cytoplasm composition, drug resistance in cancer cells and antibiotic resistance in bacteria. This has involved working closely with a major, multinational pharmaceutical company. Current applications being explored include the measurement of cytoplasmic resistance in cardiac myocyes for cardiac action potential studies, which has shown that DEP is as accurate as tissue-strip measurement, and more accurate than other methods of suspended cell measurements.
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Edit to add that Dr Labeed holds a visiting position at the University of California Irvine and has collaborators there. That is where Dr Rahim Esfandyar-Pour works.

Edit 2 : Dr Labeed is now working at the United Arab Emirates University and is now a visiting professor at Surrey according to LinkedIn
https://www.linkedin.com/in/fatima-labeed-phd-0b11b65b/?originalSubdomain=ae
and also this lab page (hosted by wixsite???)
https://yelluk.wixsite.com/my-site-1925/About
 
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Some info on the DEP device used for electrical measurement
Source : High-throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-activated cell enrichment
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5422786/
upload_2024-3-28_15-10-3.png
(A). Photograph of the DEP separation chip. The chip size is 30 mm × 20 mm. (Inset) Red section shows a close-up of the chip, showing the electrodes along the inside of the wells. The section of the chip in the image is ~5 mm square.

upload_2024-3-28_15-11-12.png
(B) Schematic of the chip, showing the three modes of DEP behavior. Cells flow from top to bottom through the wells; in the left well, cells can be seen experiencing positive DEP, are attracted to the electrodes, and held; in the center well, cells experience negative DEP, are repelled into the center of the well, and pass through. In the third well, cells experience no DEP force. In reality, cells of the same type will experience the same mode of behavior in all wells on the chip, but two different cell populations can exhibit behaviors different from each other. If one subpopulation experiences positive DEP and the others exhibit negative or neutral behavior, they can be separated.

upload_2024-3-28_15-12-48.png
(C) The chip is loaded into a fluidic cartridge comprising two parts; an upper part B contains both housing and plunger, whereas a lower part A collects the cell solution. The chip fits between the two, sealed on both sides by O rings, and is clamped together by three Allen bolts.
 
https://meassociation.org.uk/2024/0...-properties-of-blood-from-people-with-me-cfs/

Professor Robert Dorey and colleagues update on investigating the electrical properties of blood from people with ME/CFS
July 2, 2024

This jointly funded project by ME Research UK and the ME Association, Prof. Robert Dorey and colleagues at the University of Surrey and London School of Hygiene and Tropical Medicine are currently investigating the electrical properties of blood from people with ME/CFS.

The day-to-day running of the project is being carried out by Dr Krista Clarke, and she and the team have made a good start, recently updating the funding charities on their progress.

Previous research from Prof. Ron Davis showed a difference between people with ME/CFS and control subjects in the electrical impedance of their white blood cells. This indicates an alteration in the resistance of the cells to an electric current, which (while the biological implications are not yet clear) could represent a diagnostic marker for the disease.

Using blood samples from the UK ME/CFS Biobank, Prof. Dorey and team are exploring this further using improved techniques in a larger cohort of patients, including those with mild/moderate and severe ME/CFS.

They have received more than 100 blood samples from the UK ME/CFS Biobank, and, so far, 42 of these have been analysed for their electrical properties. These include samples from those with mild/moderate and severe ME/CFS, patients with multiple sclerosis, and healthy control subjects.

Findings from these groups will ultimately be compared, with the aim of understanding more about the mechanisms involved, and potentially developing an electrical biomarker for ME/CFS.
 
Medscape: Long COVID & Chronic Fatigue: The Similarities are Uncanny

"Using blood samples from the UKMEB, the researchers are now investigating this potential biomarker with improved techniques and a larger patient cohort, including those with mild/ moderate and severe forms of ME/CFS. So far, they have received more than 100 blood samples and have analysed the electrical properties of 42.

'Based on the results we have so far, we are very close to having a biomarker for diagnosis. Our results so far show a high degree of accuracy and are able to distinguish between ME/CFS and other diseases,' said Labeed."
 
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