A nanoelectronics-blood-based diagnostic biomarker for ME/CFS (2019) Esfandyarpour, Davis et al

[Sid]

Damn. That's really amping my hyperosmotic stress levels. Yoga, right?

I will throat-punch the next person who asks me, "Have you tried yoga?"

 

[Barry]

Well to be fair that seems to be what the abstract implies.

Yes, this sentence, "The test, which is still in a pilot phase, is based on how a person's immune cells respond to stress" is going to be widely misinterpreted as psychological stress, rather than biological stress.

 

[roller*]

ok, they will have checked if ppl with e.g. less blood cell deformability dont have the same signal.
(more diseases show less deformability faik)

if they say, it detects "mecfs" then other diseases must have a different signal.
perhaps there is a signal-comparison with other diseases in the study.

 

[rvallee]

I must admit I don't understand why a small pilot study should warrant such a massive media blitz. It seems to me a bit of an own goal. It leaves it wide open for the likes of Wessely to dismiss it as too early to say whether it will prove to be clinically helpful as a diagnostic test. The researchers themselves say there's a lot more steps to go through before it is proven. It could fall at any of those steps.

I'm guessing the SMC is hoping for that. It's clearly a pilot study and it needs to be understood as that. Overhyping is one strategy to be able to claim victory even in the face of defeat, where even if it's a rousing success the usual suspects will say it didn't hold up to the hype they created.

Whatever, let them stew and sweat over their career going down in flames. The science will work its way.

 

[Barry]

SW said: "is it measuring the cause, and not the consequence, of illness?"

If it really proved to be a unique biomarker, would SW's comment matter?

 

[rvallee]

I don't think this should have been pitched to the media before independent replication. This has the potential to hurt us as badly as the XMRV fiasco.

I doubt Stanford did that. The SMC had its coverage lined up and ready to go to try to overhype. That's a deliberate strategy and should not really matter. There should be no hesitation to call their choice to overhype as their own choice and fault them for it.

Meanwhile reasonable people should simply let the science speak for itself and ignore the malice from the usual suspects. Their opinion is irrelevant in the grand scheme of things.

 

[rvallee]

SW said: "is it measuring the cause, and not the consequence, of illness?"

If it really proved to be a unique biomarker, would SW's comment matter?

The only thing that matters about Wessely's opinion is that it does not matter. He likes the sound of his own voice, beyond that we can just ignore the mouth noises he makes.

 

[Simon M]

Have had a quick read through The paper and thought I'd place here before I went to bed.


They do claim that the finding is unique to ME/CFS. So those comparisons with sick controls are essential.


As I think everyone has pointed out, bigger samples are also needed. Though the Stanford piece made clear that this work is still in the pilot phase.


Patients were diagnosed "in line with Canadian criteria"; five severe, 15 moderate. Unfortunately, only five of the 20 healthy controls were age/sex matched, for some reason.


Chris made some good comments, including most of the above points, over at the Science Media Centre. Some fair points and some rather more desperate ones from the other contributors.

https://www.sciencemediacentre.org/...al-biomarker-for-chronic-fatigue-syndrome-me/


Interestingly, using whole blood (the original plan) didn't pan out. They got better results by suspending PBMC's in either their serum or their plasma. Also, best results are obtained by testing within five hours of drawing blood.


However, they still got reasonable results at room temperature per 24 hours first, or freezing in liquid nitrogen.


However, this Version of the test doesn't look like a test that would be doable in a Dr's office. Separating PBMC's from the plasma would be a lab-based test.


Using the difference between the minimum impedance value and the plateau value gave the clearest separation, but they try different measures (such as using the baseline measure rather than the minimum) and all gave clear separation between patients and controls.


Likewise, they looked at several different impedance measures and all gave clear separation between patients and controls. Which makes it look like these findings are robust (over and above the spectacular P values)


Interestingly, most of the results from moderate patients clustered towards the bottom (with the smaller separation from healthy controls (while all of the severe patients were near the top. Suggesting that severity affects results, though these are small samples.


It looks like they were originally doing the Nanoneedle test and salt stress as a way of creating PEM at the cellular level and to deplete ATP, the cell's fuel, presumably as a good model for the illness. And then discovered that the differences between patients and controls were so spectacular that they should perceive this as a potential biomarker.


Also, numerous studies had shown that adding salt was a way of stressing cells/increasing energy demand, so this wasn't just a "good idea" they had but something based in the literature.


In trying to understand what's happening biologically, they suggested that the increase in salt concentration could lead to release of pro-inflammatory cytokines and changes in gene expression.


Interestingly, they used live microscopy to look at the cells during critical stages and experiment and could see no differences between cells visually.


Okay, so they are going to perform further experiments to try to understand the specific mechanisms underlying the results. And adapting the technology so that it could be used by clinicians as any skill level.


Sorry, not been reading this thread and quicker people than me have probably posted much of this.


Night!

 

[roller*]

where is the signal for MDD major depression disorder ?

compare it with mecfs.

 

[Barry]

Interestingly, most of the results from moderate patients clustered towards the bottom (with the smaller separation from healthy controls (while all of the severe patients were near the top. Suggesting that severity affects results, though these are small samples.

So if these results are reflecting an underlying cause, then that would tentatively suggest all the patients are in a common grouping, rather than distinctly different sub-groups. But of course the results could conceivably be reflecting similar effects from different causes. Would be interesting to know if their patients had a clear distinction between moderate and severe, or if it was a broad spread of severities.

 

[Pechius]

sample of thousands or tens of thousands

Hmm... Tens of thousands?? Let's be reasonable. Nothing even close to that would be needed.

. It's quite crazy that Ron Davis himself often uses the term CFS rather than ME or CFS, given the state Whitney is in.

I don't think so. He's a high level scientist and likely doesn't want to throw around phrases which do not necessarily have much to do with reality (encephalitis).

SW said: "is it measuring the cause, and not the consequence, of illness?"

If it really proved to be a unique biomarker, would SW's comment matter?

Clearly Wessely still believes in his deconditioning nonsense, but it can easily be put to rest and I'm sure it will be. Test against deconditioned patients in bed rest, psych disease and we will have an answer.

 

[JemPD]

I find it rather hypocritical for SW to complain about the lack of comparison with other fatiguing conditons... since
i never saw him or any of the other 'unhelpful beliefs' fanclub do any study where they compared results with other fatiguing conditions... which is no surprise,since their cohort selection using oxford criteria illustrates their opinion that there is no difference between CF, CFS, fatigue from depression etc. So a bit rich to complain about that comparison not yet being done here.

They want to practice what they preach, all those studies showing correlation of rearly life trauma & CFS... with no comparison with any other illnesses so they can imply its caused by trauma.

 

[Diluted-biscuit]

I’m just too cynical after so many false starts over the years. I know one day they will find something but I’m keeping my expectations low for the immediate future.

 

[Londinium]

I think the comments above address most of my thoughts on the subject. Ideally this paper will open funding to test on other conditions; in a perfect world this would include mild/moderate ME/CFS patients, autoimmune conditions, POTS, fibromyalgia, plus anxiety and depression disorders - and sedentary controls. At this stage it's hard to know whether the result diagnoses ME/CFS or just general illness.

That all said, this is still an interesting result and whilst the saying might be 'it's the hope that kills you' I personally think that's a bit negative and that a little bit of hope here and there doesn't do any harm

(Plus as a former engineer: the engineering itself is pretty cool)

 

[Forbin]

...is it measuring the cause, and not the consequence, of illness?"

Even more to the point, can we be sure that the patient PBMC's weren't sent a newsletter telling them how to respond to the test?

 

[Wonko]

Even more to the point, can we be sure that the patient PBMC's weren't sent a newsletter telling them how to respond to the test?

I suspect it's worse than that, over many, many years (13.6B) the universe conspired, with assistance from others, namely 'evolution', to train and coach the cells to behave in exactly this way to this test.

Just to spite some people who were, in their eyes, right.

It's clearly a conspiracy by the universe, and as such it should be stricken from the record, and existence, in short it should be totally disregarded, m'lord..

 

[wigglethemouse]

I am sorry to say that I still do not understand what they did or why. I have no idea how many cells they tested or whether they even knew how many cells they were testing.

@Jonathan Edwards
This seems to be the experimental setup - measure electrical AC impedance of samples across 30nm electrodes over 3 hours. Samples were tested within 5 hours of collection - I imagine this very much limits sample size. Details from paper :

Prepared samples (Materials and Methods and SI Appendix) consisted of PBMCs (200 cells per uL) incubated in their plasma and prepared within 5 h of use.

Experimental Setup. To make electrical connections to the sensors' small (50um x 50um) measuring pads, an S-1160 probe station (Signatone) was used. A iocompatible silicone wells FlexWell Incubation Chamber (Grace Bio-Labs) was cut and placed over a sensor as a microfluidic well to contain the sample. For each experiment, 50uL of the prepared sample (SI Appendix) was injected into the microfluidic wells, and real-time electrical impedance measurements were then recorded.

Measurements. All Measurements were performed in a faradic cage to minimize 60 Hz coupling and external interference with our nanoelectronics sensors.

The nanometer-sized sensing region of the sensor consists of a 30-nm-thin oxide layer sandwiched between two 100-nm-thin gold layers.

Before the electrochemical impedance spectroscopy (Vrms = 250 mV at f = 15 KHz) (Materials and Methods), all sensors were subjected to an extensive cleaning procedure to eliminate any potential for contamination (Materials and Methods).

After reaching the baseline, we introduced a small volume (~6uL) of hyperosmotic stressor to the samples. Testing of the sample from the healthy control showed a transient decrease in impedance signal after raising the plasma's NaCl concentration to 200 mmol/L.

Hope that helps.

 

[wigglethemouse]

Thanks for the great write-up @Simon M. I'm not sure they say the finding is unique to ME/CFS - you probably meant to write something a little different here.

They do claim that the finding is unique to ME/CFS. So those comparisons with sick controls are essential.

 

[wigglethemouse]

I am sorry to say that I still do not understand what they did or why.

@Jonathan Edwards Regarding "why" take a look at the citations from the author. I suspect someone decided just to try it because they'd worked on nanoneedle technology previously.

34. Esfandyarpour R, et al. (2013) Label-free electronic probing of nucleic acids and proteins at the nanoscale using the nanoneedle biosensor. Biomicrofluidics 7:044114.
35. Esfandyarpour R, Yang L, Koochak Z, Harris JS, Davis RW(2016) Nanoelectronic threedimensional (3D) nanotip sensing array for real-time, sensitive, label-free sequence specific detection of nucleic acids. Biomed Microdevices 18:7.
36. Esfandyarpour R, Esfandyarpour H, Javanmard M, Harris JS, Davis RW (2013) Microneedle biosensor: A method for direct label-free real time protein detection. Sens Actuators B Chem 177:848–855.
37. Esfandyarpour R, Esfandyarpour H, Harris JS, Davis RW (2013) Simulation and fabrication of a new novel 3D injectable biosensor for high throughput genomics and proteomics in a lab-on-a-chip device. Nanotechnology 24:465301.
38. Esfandyarpour R, Javanmard M, Koochak Z, Harris JS, Davis RW (2014) Nanoelectronic impedance detection of target cells. Biotechnol Bioeng 111:1161–1169.
39. Esfandyarpour R, Esfandyarpour H, Javanmard M, Harris JS, Davis RW(2012) Electrical detection of protein biomarkers using nanoneedle biosensors. MRS Proc 1414:mrsf11-1414-hh04-04.
40. Esfandyarpour R, Javanmard M, Koochak Z, Esfandyarpour H, Harris JS, Davis RW (2013) Thin film nanoelectronic probe for protein detection. MRS Proc 1572:mrss13-1572-ss01-10.

Citation 38 might be relevant....
Nanoelectronic impedance detection of target cells (Dec 2013) Esfandyarpour et al
https://onlinelibrary.wiley.com/doi/abs/10.1002/bit.25171

 

[Chezboo]

SW said: "is it measuring the cause, and not the consequence, of illness?"

In other news, a Dr who shall remain nameless examines a patient who shall also remain nameless, whereupon the Dr observes a great big dirty malignant tumour, that in all likelihood signals very grave news indeed. The Dr remarks that although he can confirm there is a life threatening tumour in evidence, it can also be said that the exact cause has not been elicited. Therefore the sensible approach, according to the Dr, might go something like this “OK folks, nothing to see here, move along please.”

Edited typos