Homoeopathica September 2004

by Bruce Barwell

One of the things about homœopathy that always attracts sarcasm and jeers is the claim that solutions so dilute they contain no molecules of medicinal substance can affect human health. Scientists, even ones unsympathetic to homœopathy, become the target of comments fuelled by emotion and ignorance if they produce experimental results giving even the most oblique support to homoeopathy.

Expect a storm of ill-informed criticism to rage now publicity has been given to a paper which appeared in Inflammation Research in April this year.

Here is a summary of the paper, prepared by the publisher, Birkhäuser Verlag AG.

Histamine dilutions modulate basophil activation

Received: 11 December 2002 Accepted: 12 November 2003
Published online: 21 April 2004

Abstract
Background: In order to demonstrate that high dilutions of histamine are able to inhibit basophil activation in a reproducible fashion, several techniques were used in different research laboratories.

Objective: The aim of the study was to investigate the action of histamine dilutions on basophil activation.

Methods: Basophil activation was assessed by alcian blue staining, measurement of histamine release and CD63 expression. Study 1 used a blinded multi-centre approach in 4 centres. Study 2, related to the confirmation of the multi-centre study by flow cytometry, was performed independently in 3 laboratories. Study 3 examined the histamine release (one laboratory) and the activity of H2 receptor antagonists and structural analogues (two laboratories) Results: High dilutions of histamine (10-30 to 10-38M) influence the activation of human basophils measured by alcian blue staining. The degree of inhibition depends on the initial level of anti-IgE induced stimulation, with the greatest inhibitory effects seen at lower levels of stimulation. This multi-centre study was confirmed in the three laboratories by using flow cytometry and in one laboratory by histamine release. Inhibition of CD63 expression by histamine high dilutions was reversed by cimetidine (effect observed in two laboratories) and not by ranitidine (one laboratory). Histidine tested in parallel with histamine showed no activity on this model.

Conclusions: In 3 different types of experiment, it has been shown that high dilutions of histamine may indeed exert an effect on basophil activity. This activity, observed by staining basophils with alcian blue, was confirmed by flow cytometry. Inhibition by histamine was reversed by anti-H2 and was not observed with histidine, these results being in favour of the specificity of this effect. We are however unable to explain our findings and are reporting them to encourage others to investigate this phenomenon. [The degrees of dilution used equal 15 to 19c. – Ed.]

The British newspaper the Independent summarised this technical stuff by saying that, in summary, the study found that extremely dilute solutions can have a biological effect. Like homœopathic remedies, the solutions in the experiments were so diluted that there was no realistic chance of a single molecule of the substance remaining in the liquid.

Scientists have likened this to believing in magic. How could something that was once dissolved in a solution, and can no longer be present in that solution, still have an effect? The scientists themselves are baffled. “We are not yet able to propose any theoretical explanation of these findings,” they write. In showing that high dilutions exert a biological effect, the findings seem to break the laws of physics. Surely there must be error in the experiment; an accusation the scientists reject. “Despite searching for artefacts, we have been unable to find any,” they write.

An editorial in Inflammation Research explains why the journal published such controversial research: “The authors are unable to explain their findings but wished to encourage others to investigate this area,” it says. “It is with this spirit of openness that the journal, after submitting the paper to a rigorous reviewing process, has agreed to publish the paper.”

Understandably, the practitioners of homœopathy have seized on the findings as vindication. Peter Fisher, of the Royal Homœopathic Hospital in London and homœopath to the Queen, said the findings were nothing short of groundbreaking. “History may come to view [the study] as a turning point in the scientific controversy surrounding homœopathy,” Dr Fisher said.

“Of course further repetition is required, but it may be that this represents the holy grail of basic research in homoeopathy,” he said. Science cannot explain how such highly dilute solutions could have an effect, that is until the French biologist Jacques Benveniste came along. Working at his laboratory in Paris, Benveniste formulated the idea that water retains a “memory” of what has been dissolved in it and that it is this memory that results in the homœopathic effect. In 1988 Benveniste published a study in the journal Nature in support of his water-memory theory. He claimed his experiments showed that an ultra-dilute solution exerted a biological effect.

However, the then editor of Nature, Sir John Maddox, had insisted that he would only agree to publication if he was able to investigate Benveniste’s laboratory procedures. A few weeks later Sir John invited an American science fraud investigator, Walter Stewart, and a professional magician and arch sceptic, James Randi, to watch over Benveniste as he and his team tried to repeat the experiments.

The Nature investigation concluded that Benveniste had failed to replicate his original study. In subsequent issues of Nature, Benveniste suffered the professional ignominy of being ridiculed by arguably the most influential scientific journal in the world. As a result, the idea of memory water was consigned to the dustbin of science history, or so it was thought.

France as a country is a keen advocate of homœopathy and there were many French scientists who had not given up on the notion of investigating the phenomenon. Among them was a one-time collaborator of Benveniste called Dr Philippe Belon, who now works for a French homœopathic company, Boiron.

Belon, who fell out with Benveniste a long time ago, has investigated high dilutions for 20 years and although he works for Boiron, and has himself tried homœopathic remedies, he insists he is only interested in discovering the truth about the claims.

In the spirit of scientific investigation he organised a collaboration between four different groups in Europe who all undertook to carry out identical high dilution experiments at separate places involving separate teams of scientists.

The British end was run by Professor Madeleine Ennis, an established asthma researcher at Queen’s University of Belfast and an avowed sceptic of all things homœopathic.

In fact Ennis became involved in the project in the first place because she could not accept what some of her scientific colleagues were saying. “I told people I didn’t believe it so they said ‘Why don’t you try it?’,” Professor Ennis said.

The dilution experiments they carried out, and now published in Inflammation Research, involved histamine which is released by a type of white blood cell called a basophil. Normally basophils release histamine, and as levels of histamine rise this exerts a “negative feedback” which inhibits further release of histamine. The four teams of scientists tested highly dilute solutions of histamine to see whether they still exert an effect on basophils in a test tube. At extreme dilutions, three out of four laboratories found a statistically significant effect and the fourth found an effect which just fell out of the typical range for statistical significance. Ennis emphasised that the research does not prove that homœopathy works, nor does it even show that Benveniste was right because he had used a different test for a high-dilution effect. “The paper didn’t test homœopathy, it tested high dilutions of histamine. I know what we tested and I cannot explain the results,” said Ennis. For Belon, however, the research does at least support the basic premise behind homœopathy. “Of course it supports it, on the other hand it is not a demonstration that homœopathy works,” he said. In whatever ways the latest findings are interpreted, they cannot be ignored. The experiments were repeated by four different teams using the same experimental protocol that involved a blind code – the scientists did not know whether they were working with a high dilution solution or a control sample of pure water until the code was broken at the end of the experiment.

When BBC Horizon televised a similar attempt at replicating the same experiment two years ago, the results were negative, but scientists such as Belon believe this was trial by media rather than science by the peer-review process.

This time, with a full scientific paper detailing the precise protocol, anyone can try to replicate the findings – and replication is the essence of science. Until others repeat the work it will take a lot to convince sceptics, however.

Last year just about everyone in New Zealand with a keen interest in homœopathy saw the BBC’sHorizon programme featuring an experiment to demonstrate the reality of the power of ultramolecular solutions – and how it ended in failure to show anything significant.

The TV programme stirred quite a lot of comments, nearly all unfavourable to homœopathy, at the time. Subsequently a lot of information became available, but to a very small circle, showing theHorizon experiment was seriously flawed in its execution and contained serious errors of fact – tantamount to lies.

This information is available in detail at this website: www.homeopathic.com The following is a summary.

People with a logical turn of mind may have wondered as they watched the results of the televised experiment unfold how there could have been a mix of both positive and negative results in both the real and placebo arms of the experiment.

The explanation is that it was not an exact replication of work done by Madeleine Ennis, as was claimed by the programme, but had a major, and fatal, difference.

Wayne Turnbull, of Guy’s Hospital, who is an expert on cellcounting devices, was involved in setting up the experiment. He knew very little about basophils, histamine and so on but was a wizard at flow cytometry. He decided his counting device would work better if ammonium chloride was added to the solutions containing the cells. Ennis did not do this because she knew ammonium chloride kills basophils, thus spoiling the experiment even before the potentised histamine was added to their solution.

This is part of what Ennis emailed Dana Ullman on the matter: Speaking to Wayne [Turnbull], he really knows very little about basophils, although he is an experienced flow cytometrist. He knew nothing about the commercially available kits looking at basophil activation (there are good ones on the market) nor about the European study investing drug allergies using this methodology. As you know, I never agreed to approve his protocol i.e. assess if the study design was adequate to assess the expected differences.

He left the blood to sediment for 4 hours – this is far longer than we ever used.
He used completely different buffers including the addition of foetal calf serum.
He added in an ammonium chloride lysis step. He added in a passive sensitization step and said that the cells were used within 24 h for the assay (we would not leave our basophils hanging around for such a long time). They were left in the presence of 10% foetal calf serum (this is not a defined medium and could contain many things that might influence an assay).

I do not know where he got his anti-IgE from (this was not given in the protocol) and I did tell him that anti-IgE varies from supplier to supplier and indeed from batch to batch.

I had suggested that he tested all his donors using a dose response curve of anti-IgE – different donors respond differently to anti-IgE. It is common to have a bell-shaped curve. I suggested that he used donors that gave an activation of ca. 30-40%. I do not know how many donors he planned to use.

Given the time scale involved I am not sure if he repeated the experiments with different donors or not.

By the way – if they are trying to repeat the paper that I published on the flow ctytometric method then they should have done dose response curves for the histamine and not simply pick a few dilutions to test but that is kind of obvious.

The BBC’s experimental set-up was supervised by Professor John Enderby, of the Royal Society. He told Ullman he did not know there was any difference between Ennis’ and the BBC’s experiments. He had no idea that Turnbull changed the experiment. Ullman emailed Nathan Williams, producer of the BBC programme, pointing out the serious errors of fact that were screened.

Your defence of the Horizon programme is extremely weak, and because you stand by it without any remorse or criticism of yourself or the experimenters you used, it is obvious to me that I must continue to proceed with my formal complaint with the BBC and that I must call on the scientists who participated in the BBC experiment to acknowledge that: The BBC experiment was not a “repeat” of Professor Ennis’ work nor of any previously conducted study. (This is so obvious that even your experimenter Wayne Turnbull has acknowledged it!)
Wayne Turnbull of Guys Hospital has asserted that “consensus is vital” for this type of research, but when he sought to get consensus from Professor Ennis, he was told that his protocol was given a point-by-point listing of the differences between his experiment and Professor Ennis’.

He never received consensus for this experiment from anyone in the homeopathic community.

According to Martin Brand (the statistician for the BBC experiment), he and Marion Macey (who also conducted the BBC experiment) assumed that they were repeating Ennis’ experiment, and he reported this statement to the Royal Society. According to Brand, he and Macey had no idea that there were differences between the protocol that they used and the one that Ennis used.

The BBC experiment used 10% foetal calf serum (this is not a defined medium and could contain many things that might influence an assay). You said, ” There is absolutely no contradiction that I can see between what was stated in the programme and the way in which the experiments were carried out.” And yet, when your programme stated (twice) that the BBC experiment was a “repeat” of Professor Ennis’ work, these statements were either an error or a lie, though you somehow do not consider it either (sorry for the harsh words here, but something needs to shake you out of complacency). I do not understand your logic, nor have you presented any. As a journalist, it seems that you consider it ethical and competent to say that someone is “repeating” an experiment if there is only a vague semblance of one experiment to the next. This thinking and reporting seems to be a classic example of sloppy journalism and junk science.

Your programme was explicit in saying that you were “repeating” Professor Ennis’ experiment. And yet, the lead experimenter for this TV programme, Wayne Turnbull, has gone on record saying that, “The degranulation protocol that we use was never portrayed as a replication of Dr Ennis’s methodology.” Which one is it; was it a repeat or not? Your email below stated, “There is no evidence that any of the procedures that Wayne Turnbull followed had any adverse affect on the outcome of the experiment.” On whose authority are you saying that the changes that Wayne Turnbull initiated had “no adverse affect?” The one leading expert on this study with whom Turnbull formally consulted, Professor Ennis, told him directly that his protocol needed to be changed. After Turnbull’s conversation with Professor Ennis, she was surprised and shocked at how little he knew about basophils. Turnbull has asserted that he “stands behind” his protocol. However, considering his inadequate expertise in this area of research, his assertion has little meaning.

Further, Turnbull acknowledges that he asserts other members of the homœpathic community were “consulted in preparation for the experiment,” but neither HE or YOU have told me WHO these people were (all of the people you interviewed for Horizon have told me that they did not consent to the protocol that was used).

It is in YOUR interests to come forward now and realize that there were serious problems in the programme you aired on homeopathy . . . and to make appropriate corrections on air and at the BBC’s website.