Heh! Right now I have this stupid grin on my face, because I caught this glaring error in a published paper. Okay, it is a paper on homeopathy referenced in a godawful homeopathy website (that I mentioned in my yesterday’s write-up), but nevertheless.
Imagine, I start the introduction to a scientific paper like this:
Disease A is very important and prevalent in certain regions like blah-blah-blah. The causal organism of disease A is known to be virus X with wide seroprevalence in the said regions.
Doing cartwheel requires strength of the arms as well as of the muscles of the lower back. People who are in poor shape are afraid of doing cartwheels.
Currently, there are no treatments for virus X, nor is any vaccine available.
Therefore, this study was aimed at discovering whether doing cartwheels can eradicate the virus X.
I kid you not, my dear readers. As strange as it may seem, this is exactly how a 2010 paper that purportedly examined the effect of homeopathic belladonna on Japanese Encephalitis (JE) Virus infection starts (Ref: American Journal of Infectious Diseases 6 (2): 24-28, 2010). If you don’t believe me, read the PDF here.
The authors start with a description of the problem of JE and its geographical distribution and prevalence in Southeast Asia (which is fine), abruptly seguing into two paragraphs on the provenance of belladonna, alkaloids derived from that plant (which are not in doubt), before veering again into features of JE and vaccine prophylaxis for JE, which has been effective in several countries in reducing the incidence, commenting on the relative lack of specific antiviral therapy against JE, and finally touching upon a couple of new investigational therapies. That’s one hell of a run-on sentence, but I hope you are with me so far.
And then they end the intro with this aim statement:
There is thus a real need for antivirals that can reduce the toll of death and neurological sequelae resulting from infection with JE virus… Therefore, this study was aimed to see whether ultradiluted belladonna has a role in this infection.
Boom! … Wait, what?
Yeah. There is nothing in the introduction that remotely prepares the reader for this massive stretch from theory to an implausible study aim, this veritable leap from fact to fiction. Nothing in the introduction mentions anything about ‘ultradiluted belladonna’, nothing provides any inkling as to its purported efficacy, or indeed, as to why this particular nostrum was chosen for study.
Let me be very clear: Atropa belladonna and its derived alkaloids ≠ ultradiluted homeopathic belladonna used in this study. Ain’t quite the same.
I should also clarify that the doses used (3, 6, 30 and 200 – the authors don’t mention the designation) possibly refer to 3, 6, 30 and 200X (the ‘X’-designation is prevalent in India, the source of the preparations), which mean, respectively, the dilutions of a thousandth (10-3), a millionth (10-6) – both being fairly plausible dilutions, with reasonable amounts of the original alkaloid left – to be followed by 10-30 and 10-200 – both of which are ridiculous dilutions in which no remnant or trace of the original alkaloid can possibly remain. At these dilutions, only the solvent and dissolved impurities are around.
The experiment itself used a fairly straightforward technique of embryonic chick egg inoculation and enumeration of JE virus-induced pock marks on the chorioallantoic membrane. The virus, deposited on this membrane inside the egg, grows in situ and produces the pock marks; higher number of these marks indicates higher viral proliferation, and this is the variable authors estimated.
The test group basically checks the number of pock marks in presence of their therapy. Naturally, the appropriate controls are (a) virus control, i.e. virus without therapy, (b) therapy control, i.e. the treatment without the virus, and (c) controls for buffer (BSA in PBS) and vehicle (sterile water), treated in the same way (‘potentized’) as the therapy. The authors seem to have these covered in the methods section. So far, good.
Let’s delve into the observations. The authors contend that:
Encouraging results were obtained with all ultradiluted Belladonna 3, 6, 30, 200 preparations used in this study showing inhibition of viral growth on CAM (‘chorioallantoic membrane’). [N.B. Caveat lector: Although they mention in the discussion beneficial effects on JE in mice also, there is no mouse data presented in this paper]
Where does this extraordinary claim come from? The authors lay out their results in the following table.
As you can see, they claim significance of the homeopathic dilutions, because the number of pock marks is significantly less with their administration, than with the virus alone. It is indeed significant (but not in the way the authors intended) that they chose not to show any data from the controls, just restricting the observation to ‘no significant finding’ for the control.
The authors also chose to ignore a very vital comparison: Even in absence of the homeopathic therapy (i.e. in the virus control), the virus-induced pock marks vary significantly (p<0.001, I plugged the numbers to check) from experiment to experiment – say, between the controls for the belladonna 3 and 30 test groups, which should have been nearly identical, because conditions are the same. This inherent/experimental variability in their control groups immediately invalidates their conclusion of difference between control and test groups, because one cannot satisfactorily estimate if the observed effect is due to the treatment or simply random chance events.
The authors also fail to mention why a higher dilution (therefore, more potent according to homeopathic principles) of 6X ends up with more pock marks than a lower dilution (3X). They plot the average counts in the paper, but the graph contains no error bars, leaving no visual means to ascertain the dispersal of their test data vis-à-vis the control.
So, in conclusion, for a study that is hailed by the homeopathy website(s) as the evidence that a homeopathic nostrum works against the JE virus, the study is inadequate (only ex vivo), as well as plain darn wrong because it draws wrong conclusions stemming from a major logical flaw: confirmation bias. The authors saw in the observations only what they wanted to see, and didn’t analyze the data critically. This study is sloppy, and I honestly wonder how it even passed peer review to be published in the American Journal of Infectious Disease (admittedly not a journal with whose academic standards I am familiar).
But what irritates me the most – perhaps a tad irrationally – is that the primary authors are from my own city and are employed in prestigious and historic institutions for clinical and basic research. To me, this only reflects the sad reality of the inexplicable iron grip on the general populace, including – shamefully – scientists, that various pseudoscientific, nonsensical, faith- and superstition-based modalities, such as homeopathy, continue to exercise. Geek Nation, anyone?