When this appeared in my inbox yesterday, courtesy of Richard Saunders (who was kind enough to ask me if I was sitting down first) I initially *facepalmed*, then sought about getting the original paper where this apparent break through research had appeared (thanks to @xtaldave for the full text).

The paper that had apparently found homeopathy to be as effective as chemo for breast cancer (according to Homeopathy Plus!, yes those guys), was published in the International Journal of Oncology* and entitled “Cytotoxic Effects of Ultra Diluted Remedies on Breast Cancer Cells.” (Click the link for the full pdf of the study).

The paper examines the effects of ultra-dilute remedies (read:homeopathy) on the induction of cell death in two cancer cell lines (commercially available MCF-7 and MDA-MB-231) and one immortalised control cell line, (HMLE).

The authors use several remedies already in use for the treatment of human breast cancer developed at the P. Banerji Homeopathic Research Foundation in India, Kolkata;

Carcinosin, 30C; Conium maculatum, 3C; Phytolacca decandra, 200C and Thuja occidentalis, 30C (for an explanation of how dilute these remedies are see here).

All remedies were diluted in 87% “extra neutral alcohol” and succussed, including the alcohol used as the control solvent.

The authors analysed each remedy with high performance liquid chromatography (HPLC) to look for differences, then measured cell death in cell culture in response to increasing doses and increasing time of incubation with each remedy. These measures include the MTT assay for cell viability, Annexin V and PI for apoptosis, FISH for DNA breakage and Western blots to measure activation of cell cycle proteins.

Technically the paper is quite well written. The problems lie with the data. And these problems are so massive, I wonder how they got by the reviewers. I don’t know whether they were dozing when they reviewed this paper, but I could immediately see some big fat gaping holes in their results.

First up a few pointers;

“The experiments were conducted in triplicate and repeated at least twice in each case of remedy”

This would not get past me. It is accepted scientific convention that experiments are done at least three times (not two) and also in triplicate, giving you a final “n” number of 9. These studies were done in cell culture, meaning there is plenty of material for experiments to be repeated as many times as you wish. So why were they only done twice? Three is convention because it gives the study more statistical power.

Ah statistics, huh?

There is a distinct lack of statistics in this paper, by which I mean there are none at all. As my friend Jo said; “Nary a p-value nor a confidence interval to be seen”. Which begs the question, how can you get a paper accepted in a peer reviewed journal without doing an statistical analysis?

Really? No, I mean REALLY? This is why I suspect the reviewers were dozing or drunk.

And by not doing any statistical analysis, you can not make any statements about whether the treatments are different to each other. Statistics uses algorithms to calculate mathematical differences with a degree of confidence (usually 95%) so that we don’t rely on visual interpretation, which is notoriously unreliable. But this doesn’t seem to have bothered these authors, or the reviewers.

So let’s look more closely at the results.

Firstly the HPLC.

Oh wait a sec, there are no results shown for HPLC. And neither do the authors say “no results shown”. They just make some rather confused statements about what they think they saw and move on. What?! I need to see the chromatograms. What possible reason could they have for not including this data, especially when they go on to describe it so badly in the text.

“All four remedies had very similar HPLC chromatograms to each other, with only trace amounts of limited number of peaks. They were not significantly distinct from the solvent and they lacked the distinct peak seen in the solvent.

So, this means that all the remedies were the same, ie. no different to the solvent and no other peaks indicating any ingredients. But then they contradict themselves by saying that the remedies did not have the solvent peak? Fail.

And then;

“The chromatogram of the untreated and treated solvents appeared identical, indicating that succussion did not cause chemical changes in the solvent.”

Okay, but don’t some homeopaths claim that succussion does have an effect on the chemical structure of the water/solvent? Isn’t this how they explain that homeopathy works? I can only guess SINCE WE CANNOT SEE THE CHROMATOGRAMS, but what you are saying is the remedies and the solvent were exactly the same, meaning they are solvent.

No surprises there.

What about the cell death studies?

So let’s look at the cell death studies since these constitute the crux of the study’s aims. That is, to determine if these ultra dilute remedies can induce cell death in cancer cell lines.

So here we have results for all three cell lines, two cancerous and one control, and they are all treated with a control (the 87% alcohol solvent) or the remedies and death measured by MTT assay. Here’s how they describe it in the text;

“Interestingly, the inhibitory effects on cell viability of the remedies in both the MCF-7 and MDA-MB-231 cells were distinctly greater for each of the doses tested than those seen in cells treated only with solvent.”

Which translates as the treatments killed the cells better than the solvent alone. Okay, so it looks like it did when you eye ball the histograms, but you have no evidence for this – you didn’t do stats, therefore you cannot say this! Sheesh, where did you learn to write science?

But why not keep the fail going;

“MCF-7 cells were found to be more sensitive to all four remedies than the MDA-MB-231 cells”.

Again no statistics, so this statement cannot be confirmed. When you do science properly and you run statistical analysis, you are entitled to say, “MCF-7 cells were found to be significantly more sensitive to all four remedies than the MDA-MB-231 cells”.

Unless you’re these authors, then you just get a great big FAIL stamp on your work.

Also note that they state that the control treatment (that is the solvent) also induced cell death in all cell types;

“As shown in Fig. 1A, the solvent reduced the viability of all three cell types; the overall reduction in cells at different doses of solvent was about 30% for MCF-7, 20-30% for MDA-MB-231 and 20% for HMLE cells.”

Ummm, hold on a sec.

This is your control treatment, which means it should not be causing cell death. It is designed to be inert, functioning as a carrier of your treatment, in order that you can measure the impact of the treatment alone. If your solvent or vehicle is killing your cells you have a fundamental problem. You need to go back to the drawing board and find a different solvent to deliver your treatment.

This is a very big problem right here.

If the cell death induced by the solvent is significant, then the rest of the paper is worthless. But because there are no stats here, there is no way to tell if death by the solvent is significant. According to the above statement, the alcohol killed ~30% of the cancer cells compared to no treatment at all. Although this effect was increased when the treatment was present, there remains a large problem with your model if your solvent is killing the cells.

Perhaps this explains why there are no stats in this paper? Because they may in fact show that the “inert” solvent also significantly kills the cancer cells? Once again, there is no way for me to know this without access to the raw data, or the statistical analysis.

Man, how the hell did this embarrassment get accepted?

Well now that I have revealed a fundamental flaw in this tripe I have lost the will to continue. There is much more fail herein however, I mean we are only at Figure 1 remember.

So I will cover just a few more things that are also glaringly obviously wrong with this paper, then I will send a large bottle of 87% alcohol to the editorial board and encourage them to keep up the good work of smiting the peer review process and science in general.

General lack of quantitation of results in this paper.

Figure 3, excerpt from Frenkel et al., showing damage to DNA.

Figure 3 (left) shows fluorescent microscopy data for DNA breakage as measured by FISH assay. But where is the quantitation of this data?

The authors show a representative image for each treatment, and this is usually acceptable if you then measure large numbers of cells and report on such changes with numbers (see below).

Dunlop et al., in press. Panels are representative DAPI/FITC overlay images of at least 10 images taken from triplicate wells. Histograms are mean + SD of 3 independent experients, incubations in triplicate, n = 9, p < 0.001 1-way ANOVA. Tukeys post-hoc analysis.

Further, even in the fluorescent images the authors only show a maximum of 14 cells. What the hell can you glean from 14 cells? They even say;

“At least 200 cells from treated and untreated samples were analyzed for mitotic index and telomeric DNA signals with a Nikon Eclipse 80i microscope equipped with fluorescence attachment and a Photometrics CoolSNAP HQ2 monochrome digital camera.”

So where is this data? Not in this paper, I can tell you that.

Next up Western blots.

Then they move onto Western blots. Actually this looks like the most resolved part of the paper. They have normalised everything to beta-actin as convention goes, and they have indicated the time of exposure to treatments. But as I mentioned earlier, if their control treatment 87% alcohol, is killing cells, then what can we glean from this data? Well not much except the effect could be an additive effect of the alcohol and treatment. There is no way to differentiate the impact of the treatment versus the control.

Flow cytometry, not quantitated either.

Sigh. I spend most of my days doing flow cytometry, so I am pretty familiar with how it works and what are the accepted ways to present the data. This is not one of them.

Figure 5, excerpt from Frenkel et al.,

The assay they use (Annexin V and PI) is a common one and I use it often. Standard procedure is to count ~10,000 cells for each condition, then plot your results on a graph, like this (see left below).

Dunlop et al., in press. Flow cytometry analysis of lysosomal destabilisation in THP1 human monocytes with acridine orange as a probe. Mean & SD of three independent experiments, incubations conducted in triplicate (n = 9), ** p < 0.01, *** p < 0.001, 1-way ANOVA, Tukeys post-hoc analysis.

You might also show your scatter plots as they have done above, as a nice visual demonstration of how the cells respond to the treatment, but this is not quantitation.

I’m going to stop there. I won’t even bother dealing with the discussion and conclusions, because by my analysis, they are based on flawed data.

One thing I will say about Homeopathy Plus! yelling “Homeopathy as good as chemotherapy for breast cancer” is not a conclusion you can draw from this study.

For all the reasons I have addressed above as well as the really obvious point that these studies were conducted in cell culture. This is a very different situation to a whole animal.

Cells bathing in a bath of homeopathy is very different to the processes which occur in vivo, for example the treatment must survive the low pH of the stomach, cross the gut, escape metabolism in the liver and get to the site of the cancer then do it’s job. This is a very complex process and very difficult to control. Studies in cell culture can provide data about the mechanism of action of a compound, but rarely do they relate to the processes in a human.

Never extrapolate results from a culture dish to a whole animal. You will undoubtedly be wrong and look like a fool.

Ooh, did someone say Homeopathy Plus!?

Listen to Fran Sheffield from Homeopathy Plus! talk about how homeopathy works here (mp3, 3:19).

*The International Journal of Oncology, impact factor 2.234, fail factor 10^23.

Her story was featured on Channel 7s Sunday Night and I previously blogged about it here.

Well, today I was informed that tragically Tamar succumbed to her cancer on November 12th, 2009 in San Salvador where she was undergoing treatment with natural therapies, including hours of mud-wraps. There is little information on this at the time of going to press, except to say that my source tells me they contacted a journalist involved in the story, who confirmed this was the case and that Tamar passed in hospital. Google searches are also turning up very little, however I did find a comment from someone on a news site, dated December 31 st 2009, stating

“Tamar died a month or so ago in el Salvador. Towards the end the mother was taking her for chemo treatment but it was too late. Her death was reported on the news here two days ago – the hospital in el salvador has confirmed it.”

According to the 3news.co.nz story from October 12th 3009, doctors in Perth

“say a seven week course of chemotherapy will give Tamar a 50 to 60 percent chance of survival. Without it she will suffer a long and painful death.”

But her parents are devout Christians, and say God will decide her fate.

“I don’t want to lose my daughter, but it is God’s decision,” says Tamar’s father Trevor Stitt.

“Who am I to fight against it?”

Trevor also left a comment on the same website stating;

14 Oct 2009 8:17p.m.

Hello to all of you, yes I am Tamar’s Father.Channel 7 came into our lives to supposedly portray 2 sides of the story, sadly they’ve lied and left out half the truth that they filmed. They were supposed to interview people (Doctors)and natural healers who’ve cured people of cancer successfully, but “pulled the pin” on the interviews because it would topple their one-sided story.They also ommited to say that I trained at Guy’s Hospital, London and worked as an Operating Dept Practitioner in Anaesthesia in the UK for 10 years before coming to Oz in 1998,worked at Royal Marsden cancer hospital in London and so have quite good research engines at my disposal. I have seen “clay wraps” cure my Tamar of asthma at 2 yrs when PMH only offered steroidal inhalers, she’s never had an attack since.If 7 had’ve told the truth about what they saw without overdramatising the San Salvador situation and others for sensationalism, then maybe folks would know more of what we are doing for Tamar, who IS getting a little better each day.I have to say I don’t really know who you are Josie, you say you know us,do you have another name?
I guess the struggle is just beginning (and I don’t speak about Tamar’s condition here)those “orthodox”people will not be happy, google Roy Rife, see what happened to him.Remember Patch Adams? (a true story)didn’t fit in to orthodox practice, too happy,he had a proper bedside manner (taboo in today’s conventional medicine). “There are none so blind as those who will not see”- with the eye of faith you don’t only see, you begin to LIVE!Thanks to All who support us and also to those who don’t.

This is a terribly tragic case, especially if it is true that Tamar’s mother did eventually take her for chemotherapy, as suggested by the comment above. Even more so, because her father apparently has a medical background including working “…as an Operating Dept Practitioner in Anaesthesia in the UK for 10 years before coming to Oz in 1998,worked at Royal Marsden cancer hospital in London……so have quite good research engines at my disposal.” (see above). Despite this, Tamar’s parents still chose to seek out alternative remedies as oppose to those supported by evidence.

Cases such as these where religious beliefs and faith in alternative medicine result in (often) avoidable deaths are tragically, not uncommon. It is also important to consider the desperation and vulnerability of a person faced with the imminent death of their daughter, therefore one can sympathise that you might be inclined to try anything. Similar to my Mum’s friend who passed 6 months ago from cancer also, but in her final months fell victim to all kinds of useless therapies including hair analysis which I have previously blogged about here and here and vega testing.

It reminds me of a quote from an essay written by Michael Baum and Edzard Ernst in the book “Healing, Hype or Harm” about ethics in alternative medicine;

“Even treatments which by themselves are not harmful, will become life threatening if they are used as an alternative to effective treatments of serious diseases.”

Cancer Victoria produces a booklet warning cancer patients about “quack” therapies. You can read about it here.

Several sources have advised me that there will be a follow up story on Today Tonight this coming week.

—

Thanks to Tony for letting me know about this. Please see Tim Farley’s comprehensive website What’s the Harm? for more such cases.

This story was covered on Channel 7s Sunday Night programme, the same one that covered the death of Dana McCaffery and the anti-vaxers back in April. You can read my previous blogs about this here and here.

In this case, a 10 year old girl Tamar, was recently diagnosed with liver cancer which required immediate and aggressive chemotherapy. But her parents have shunned conventional treatment in favour of “mud” therapy. A team of oncologists at Princes Margaret Childrens’ Hospital advised that a seven week course of chemotherapy would give Tamar a 50-60% chance of survival. Despite the treating hospital pleading with the parents to consent to chemotherapy, eventually seeking the involvement of the WA legal system, the parents fled Australia to El Salvador.

Their preferred treatment is tea made from herbs, and red clay gathered from around the hills near their house in El Salvador. Tamar’s mother says that “Clay is basically the right medicine for any kind of illness, (it can cure) anything“. She went on to say, “..it dries up anything that is causing the illness in your system“. Twice a day, every day for a minimum of three hours, the clay is wrapped around the girl’s torso.

Her father says, “I don’t want to do this to my daughter but if it’s God decision then so be it.” He claims his daughter doesn’t want to lose her hair or get sick from the chemotherapy and has seen the research for herself, so she is therefore capable of making a decision to use natural therapies. He also claims she is eating normally and has gained weight. The parents claim the proof for the efficacy of mud therapy therapy is a book “written by a doctor” about the curative effects of herbs.

You can watch the full video here.

Thursday 23 April, 2009

People who falsely promote and provide unethical, fraudulent and potentially dangerous treatment for cancer are the focus of three significant documents launched by the Cancer Council today.

A book titled ‘Complementary and alternate cancer therapies‘, a fact sheet to help patients and their families make decisions about therapies and another fact sheet for medical professionals with tips on how to help patients avoid such practices has been produced with funding from the State Government of Victoria, Department of Human Services.

‘Many people with cancer use some sort of complementary therapies for legitimate reasons such as maintaining hope and staying in control of their situation,” Dr Amanda Hordern, Deputy Director of the Cancer Information and Support Service said.

“Health professionals are the best people to help people make a safe treatment choice so we have gone to great lengths to publish some guidelines to assist them with this.

“Targeting sick people with unproven alternative therapies can be fraudulent and may have a huge effect emotionally, physically and financially on the person with cancer and their family.

“The new information has been developed to help people recognise which therapies may be helpful and which may be making false claims. This is an important aspect with many complementary and alternative therapies proving to be a risk for some cancer patients,” Dr Hordern said.

Associate Professor Michael Jefford, Clinical Consultant to the Cancer Council said people usually used these treatments hoping to get better.

“People are hoping to shrink or even cure their cancer, deal with side effects, or just improve how they’re feeling,” Associate Professor Jefford said.

“Sadly, people with cancer rarely discuss the use of complementary and alternative cancer therapies with their cancer specialist.

“Doctors are particularly concerned if people use unproven, alternative treatments instead of standard treatment options. They need to discuss all options with their doctor and then make an informed decision on alternative therapies,” he said.

“A major worry is when people with curable cancers don’t get the treatments that could cure their disease.

“It’s about access for patients to the right information and ensuring that they come to the best possible decision that will give them the best possible chance of dealing with their illness.

“Cancer Council Victoria has developed some really good information about these treatments.

“The Cancer Council Helpline has easy access to information about a lot of complementary and alternative medicines. Also, the Cancer Council is training health professionals to talk about complementary and alternative medicines with their patients,” he said.

Cancer Council Victoria keeps statistics on all cancers in Victoria and over the past 20 years survival rates have increased year on year. Detailed stats can be found at www.cancervic.org.au.

The booklet and fact sheets are available free, through the Cancer Council Helpline, 13 11 20, and are also available online.

To interview Dr Amanda Hordern or Associate Professor Michael Jefford please contact Tim Coghlan on 9635 5262 or 0431 150 365.

I have previously blogged about cancer quackery here.