Assessing cellular energy dysfunction in CFS/ME using a commercially available laboratory test, 2019, Morten, Newton et al

One way to look at the situation is that patients were being sold an expensive test that isn't fit for purpose. Maybe the replication attempt is flawed and the test works but it's not a good sign that there is disagreement. The possibility that the test isn't fit for purpose must be seriously considered.

It's painful but if this kind of questioning didn't occur, real progress would be delayed even further (there would at best be an illusion of progress).

It's better finding out that something doesn't work after a few years than finding out after 20.

That any replication attempts are being made in general is a good sign. It would be worse if there were no attempts.

 

I hope this isn't just left as it is now. This sort of discrepancy does occur quite a lot in science. And to the outsider it can seem frustrating because one naturally asssumes that one party is correct and the other is wrong. Sometimes, it's not that simple - especially when it comes down to methedology, which often isn't definite, but a series of approximations and assumptions, with 'best' practice rather than 'correct' practice.

 

when we did the tests, blood had to be at Biolab for 9am the next morning, with a recommendation that blood be drawn in the previous afternoon to minimise timeframe. It was a We good bloods drawn at 4.10pm and was in the post at 4.30pm.
There is, I believe, a viability test done re blood quality and if not suitable ( assuming a level pf degradation) the test is not run.
The curious bit is the accurate correlation with severity.

Norman Booth was passionate about replication. Given the current situation, I agree with @wigglethemouse - get the labs working together to bottom out/ test differences - it could be a very positive thing for an illness where there are so few positives.

 

Except that in my experience that sort of artefact happens all the time in lab work. It may not be down to one thing. Systematic errors in research labs are everywhere.

 

I suppose it's possible but I wonder as to mechanism.
E.g. I was wondering if Mc/My ran a test on controls vs ME and did the ME batch after the controals, thus occasioning ME to produce worse mito function due to delay, or whether they got lots of ME diagnosed samples from clinics, kept some longer than others due to circumstance (workload, postage etc,), found they were worse - due to delay -, then "confirmed" them as ME while fresher samples with better function were labelled as not or less severe ME. There must have been some means for producing this artefact, if that is what it is.
Perhaps this is covered in the critical paper but I couldn't take it all in.

 

The authors of the present paper have some data that suggests cells change over time. It's hard to understand the technical details but the meaning is clear.

If the Myhill group used as controls samples that were fresh, while patient samples were shipped by mail and at least 24 hours old, that could plausibly produce consistently different results between sick and healthy patients.

 

Such questions need to be asked.

 

Are you familiar with supervising research assistants and doctoral students in a lab? Or with colleagues running experiments in a lab?

 

I'm reading one of the papers on the test now https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2680051/

They write

It doesn't specifically say that the delay between blood draw and testing was the same for both groups. If the authors believed that it wasn't relevant, they wouldn't consider there to be any differences in the processing even with different delay in both groups.

Such a hypothetical difference in processing wouldn't be able to explain this nice correlation between illness severity and their mitochondrial energy score.

 

Well hopefully they will send in a response to the publishing journal so that it is published alongside this paper. Not only is that the way, as I understand it, the process is meant to work, but it also increases the chances of someone else being interested in investigating this further.

ETA: a missing "s" bugged me enough that I had to edit it in.

 

The trend between CFS Ability scale and mitochondrial energy score is intriguing, and the Ability score was blinded to testing. Perhaps there is something there, and this should be explored further but the test is a long way from standardisation and commercial application.

What would also be good to see is a plot of sample processing time against MES.

Or, a new experiment with control and patient samples repeatedly tested over incremental time points, where the venepucture was done on site, so the first time point is zero.

 

McL is a senior scientist and it is reasonable not to expect such errors, though I accept your implication that in real life labs they do occur. Timing problems may be most likely but cannot be simply assumed and I think the points and questions should be put to Dr McL-H, which I hope he would respond to. The critical paper may be right as to results but wrong as to what went wrong at Akumen and that should be elucidated.
I am not nor have I ever been involved in lab work. I express my position without expertise, have a go at guessing what might have gone wrong in line with the critical paper's surmise, ask for questions to be put to Drs My/Mc and hope my inexpert knowledge will then be advanced with everyone else's. If I express no doubt, I get nowhere. I am happy with that and it is the approach of many intelligent sufferers - the little people must fight for themselves and not be shy about showing annoying naivety, if the aim is to learn - because the big uns have let us down too often and disagree among themselves anyway.
A fair criticism of work can be accompanied by an unfair or inaccurate surmise as to what went wrong. There is nothing wrong with criticising the criticism.
Those who are familiar with lab work may have better ideas as to the mechanism of Mc/My's assumed error which I hope will be discussed. What will be said if McL has records showing timing was not the issue?

 

OK, that is useful to know. I accept your viewpoint from that position.

It might be reasonable but sadly it isn't. Unconscious cherry picking bedevils all work of this sort. As Richard Feynman said, the easiest person to fool is yourself. That is why we need replication.

The graph of mitochondrial score against severity might look convincing because it is so clear cut. However, from my perspective as a lab biologist this graph shouts artefact at me. The scatters don't look right to me. If the scores in the severe cases were really that low then it should be a trivial exercise to identify what is wrong and it should be replicable by any lab. If the scores reflected some general metabolic problem rather than just some quirk of neutrophils (which would be the interest of the study) then we would expect the person to be in intensive care with little chance of survival for 24 hours. It just does not make biological sense to me.

 

The mitochondrial energy score is constructed from several different measured parameters. Maybe that's why it doesn't look like a natural thing, because it's a human construct.

Edit: I'm inclined to agree that very low mitochondrial energy scores are suspicious because it implies that in the respective patient, at least one step of ATP generation is extremely impaired. That's how I understand it anyway.

 

Agreed

From my position of ignorance your point sounds reasonable (no sarcasm intended btw).

That's the kind of issue which it would be interesting to elucidate with McL/My. I have myself always wondered to what extent any findings on compromised mitochondrial function - if valid- reflect a local or systemic problem. Another problem is that if mito compromise is really found in a systemic illness, which is not classically an illness of the mitochondria, we still don't know how to interpret such findings.

I don't personally know whether My/Mc ever presented their findings as a diagnostic test, but phrases such as "mitochondrial not psychological" were bandied about, which in itself is a contentious statement, assuming not only reflection of test result in symptoms - now at issue- but also implying that mitochondrial is an antonym of psychological - easy meat for Sharpe, Wessely etc - then it may be a quick call to the SMC to show how naive we are about mental illness and hence unable to recognise our own "mental" illness...........

 

ETA: This is wrong, so don't waste spoons reading it. (Moved this upfront as a civil service. )

Also, the numbers are multiplied together not averaged, so you'll always end up with a low number and this may exaggerate the effect of delayed testing.

Let's say you score almost full function for everything at 0.8 (out of 1.0). So let's convert that to an MES. 0.8^5 is just under 0.33 (~30% on the Bell Disability Scale), which is severe ME.

If you think of it that way, only slight deviations from the maximum score give you low results. 0.7^5 is just under 0.17. Therefore most patients must actually be scoring somewhere between 0.9 (averages at ~60% on the Bell Scale) and 0.8 (~30%, as above). Few will score lower than 0.75 (~0.24/20% Bell Score).

Could that be an artefact of delayed sampling time or lab error? It's plausible. Patients are already mobility impaired and have other factors potentially interfering with or complicating sampling. So it may take longer to get the draw, or you might have to have blood drawn at home. The more impaired the patient, the more this may be an issue (remember that blog by Anil where it took ages to find a vein for an IV drip?).

That extra time before running the tests on samples might be enough to see a drop from an average of 1.0 in all measures to 0.9 or 0.8. Especially if, as the MEA study shows, the samples seem to have lower scores the longer they're left.

ETA: This could, in theory, be tested rather easily.
ETA2: This is wrong. I misread the y axes.

 

These are the five factors which as I understand are multiplied and then divided by 0.182 to obtain the mitochondrial energy score (so your calculations would be incorrect). Here are these five factors, together with the CFS ability Scale scores.

 

0.35 and 0.11 scores

 

Ah, my mistake. I missed the division and didn't realise the numbers didn't all run 0-1. Is the first score up to 3? And another looks like it tops out at 0.8? I can't read it too well. The others look like they're still 0-1. So it looks like you can safely ignore me.

But couldn't the multiplication still magnify any difference? There are still a lot of times you're multiplying by less than one.

 

It is a couple of years since I read the papers, but from what I recall ( correct if wrong), there was also the identification of potential subgroups related to ox phos mechanisms.

My daughter fell into one of these, where ATP was being chucked out of the cell faster than it could be replenished - didn' t have a clue what it could be then , but now it sounds like purinergic signalling.