That’s what you will read if you check out discussions of a new paper, Influence of pediatric vaccines on amygdala growth and opioid ligand binding in rhesus macaque infants: A pilot study, by L. Hewitson, B. Lopresti, C. Stott, N.S. Mason, and J. Tomko.
If you are wondering, yes, that is the same Laura Hewitson of Thoughtful House who first presented the “monkey studies” at IMFAR a few years back. And, yes, that is Carol Stott, formerly of Cambridge. And, yes, this is a part of the Wakefield-team “monkey studies” which has had such a checkered history.
What is this new study about? Well, here’s the abstract:
This longitudinal, case-control pilot study examined amygdala growth in rhesus macaque infants receiving the complete US childhood vaccine schedule (1994-1999). Longitudinal structural and functional neuroimaging was undertaken to examine central effects of the vaccine regimen on the developing brain. Vaccine-exposed and saline-injected control infants underwent MRI and PET imaging at approximately 4 and 6 months of age, representing two specific timeframes within the vaccination schedule. Volumetric analyses showed that exposed animals did not undergo the maturational changes over time in amygdala volume that was observed in unexposed animals. After controlling for left amygdala volume, the binding of the opioid antagonist [11C]diprenorphine (DPN) in exposed animals remained relatively constant over time, compared with unexposed animals, in which a significant decrease in [11C]DPN binding occurred. These results suggest that maturational changes in amygdala volume and the binding capacity of [11C]DPN in the amygdala was significantly altered in infant macaques receiving the vaccine schedule. The macaque infant is a relevant animal model in which to investigate specific environmental exposures and structural/functional neuroimaging during neurodevelopment.
Basically, they took 16 monkeys (rhesus macaque or Macaca mulatta). 12 of them were given vaccines in a schedule intended to mimic the U.S. vaccine schedule of the 1990’s, including thimerosal (which was added). 4 were given saline injections (controls). MRI scans were taken. “Time One (T1) at approximately 4 months of age and Time Two (T2) at approximately 6 months of age.”
From the abstract we see that they found that the “Volumetric analyses showed that exposed animals did not undergo the maturational changes over time in amygdala volume that was observed in unexposed animals.”
In other words, the amygdala volume was different from the controls at T2 for the monkeys given vaccines.
Want some more detail? Well, in regards to the right amygdala:
For the exposed group there was a nonstatistically significant increase in right amygdala volume over time (P=0.16; Table IIa). For the unexposed group there was a significant drop in right amygdala volume over time (P<0.0001; Table IIa).
Read that again. Did they just say that a piece of the brains of the control animals shrank between 4 months of age and 6 months of age?
They did. That’s what their data show. It seemed so odd to me that I double (and triple) checked. I’m sort of visual in how I like to take in data, so here is Figure 4(A) from the paper. This shows the left amygdala size for the two times (T1=4 months of age and T2=6 months of age). I’ve added text to the graph. It is in red so you know what I added. (click to enlarge)
The dotted lines are for the “exposed” animals. I.e. those vaccinated. The solid line is for the “unexposed” animals. See how at T1 they have amygdala sizes that are about the same size? But at T2 (2 months later) the amygdalas of the “unexposed” animals have shrunk, while the amygdalas of the exposed/vaccinated animals grew a little.
I’m not a primate expert, but it bothers me somewhat to hear that a piece of the brain might shrink. I would expect in my own naive way that pieces of the brain would grow as monkeys mature, so I decided to check: has anyone looked at amygdala size in Rhesus Macaques as a function of age? It turns out there is a paper just out in 2009, “Maturation of the Hippocampal Formation and Amygdala in Macaca mulatta: A Volumetric Magnetic Resonance Imaging Study” by Christa Payne et al. from the University of Texas and Emory University. They also were working with small numbers (11 in the male group). Here is Figure 6(A) from that paper:
FIGURE 6. Modeled developmental trajectories for left (thick, thatched lines) and right (thin, solid lines) amygdala volume in males (A) and females (B). Actual volume measurements are represented by filled (left hemisphere) and open (right hemisphere) symbols.
One line is for the left amygdala, and one for the right. Same with the datapoints, the filled are for one side, the hollow for the right. But the basic idea is clear–the amygdala grows with time in monkeys, not shrink. Yes, seems obvious, but I had to check.
How could the Hewitson paper report that the control monkeys have shrinking amygdalas? One possible answer: too few monkeys in the control group. There is a lot of scatter in the amygdala data from the U. Texas paper. If someone has only a couple of datapoints, they might get some strange results.
The Hewitson paper had really small numbers:
“A complete set of MRI data at both T1 and T2 were obtained from 9 exposed and 2 unexposed animals.”
But, wait, remember above? Weren’t there 4 monkeys in the control group and 12 monkeys in the vaccinated group? What happened to the other 2 of the control subjects? There weren’t many to begin with but half of the control group are missing in the data? What’s the reason for that? No, that’s a real question which I can’t find answered in the paper: what happened to the two other controls?
This paper is generating quite a bit of interest in places like the Age of Autism blog. Unfortunately for them, this paper is not the genie getting out of the bottle. Just another low quality paper. Just another 16 monkeys giving their lives for nothing.
Left Brain Right Brain 
It is very hard to not get snarky on this one, but this is one bad study. I’m stunned it got published. There is a reason why it isn’t in a first-line (or second line) journal.
They have 2 control animals. Two. They had 4 but they just don’t report on half of them?!? The amygdalas are shrinking in their control animals with time?!?
What a waste of time, money and primates.
http://www.sciencebasedmedicine.org/?p=100 Not a new study; they finally found someone to publish it. And it’s something that had Wakefield’s name on it in 2008 when it was first presented.
thanks for that KWombles.
It isn’t the same study. Well, it is and it isn’t. Take a look at the post that follows this one (written thanks to your link). They lost the data on some of their monkeys and the results turned 180 degrees around.
On a later post at SBM, a Heather actually copied the presentation on this study along with the others, and Gorski responded he’d dealt with a couple of the monkey studies at the link I provided above. I copied and pasted her copy/paste of the abstracts of this particular one and the other two on a post I was apparently working on around the same time you were doing this one.
Here is the abstract in 2008:
Pediatric Vaccines Influence Primate Behavior, and Amygdala Growth and Opioid Ligand Binding
Friday, May 16, 2008: 5:30 PM
Avize-Morangis (Novotel London West)
L. Hewitson , Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA
B. Lopresti , Radiology, University of Pittsburgh, Pittsburgh, PA
C. Stott , Thoughtful House Center for Children, Austin, TX
J. Tomko , Pittsburgh Development Center, University of Pittsburgh, Pittsburgh, PA
L. Houser , Pittsburgh Development Center, University of Pittsburgh, Pittsburgh, PA
E. Klein , Division of Laboratory Animal Resources, University of Pittsburgh, Pittsburgh, PA
C. Castro , Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA
G. Sackett , Psychology, Washington National Primate Research Center, Seattle, WA
S. Gupta , Medicine, Pathology & Laboratory Medicine, University of California – Irvine, Irvine, CA
D. Atwood , Chemistry, University of Kentucky, Lexington, KY
L. Blue , Chemistry, University of Kentucky, Lexington, KY
E. R. White , Chemistry, University of Kentucky, Lexington, KY
A. Wakefield , Thoughtful House Center for Children, Austin, TX
Background: Macaques are commonly used in pre-clinical vaccine safety testing, but the combined childhood vaccine regimen, rather than individual vaccines, has not been studied. Childhood vaccines are a possible causal factor in autism, and abnormal behaviors and anomalous amygdala growth are potentially inter-related features of this condition.
Objectives: The objective of this study was to compare early infant cognition and behavior with amygdala size and opioid binding in rhesus macaques receiving the recommended childhood vaccines (1994-1999), the majority of which contained the bactericidal preservative ethylmercurithiosalicylic acid (thimerosal).
Methods: Macaques were administered the recommended infant vaccines, adjusted for age and thimerosal dose (exposed; N=13), or saline (unexposed; N=3). Primate development, cognition and social behavior were assessed for both vaccinated and unvaccinated infants using standardized tests developed at the Washington National Primate Research Center. Amygdala growth and binding were measured serially by MRI and by the binding of the non-selective opioid antagonist [11C]diprenorphine, measured by PET, respectively, before (T1) and after (T2) the administration of the measles-mumps-rubella vaccine (MMR).
Results: Compared with unexposed animals, significant neurodevelopmental deficits were evident for exposed animals in survival reflexes, tests of color discrimination and reversal, and learning sets. Differences in behaviors were observed between exposed and unexposed animals and within the exposed group before and after MMR vaccination. Compared with unexposed animals, exposed animals showed attenuation of amygdala growth and differences in the amygdala binding of [11C]diprenorphine. Interaction models identified significant associations between specific aberrant social and non-social behaviors, isotope binding, and vaccine exposure.
Conclusions: This animal model, which examines for the first time, behavioral, functional, and neuromorphometric consequences of the childhood vaccine regimen, mimics certain neurological abnormalities of autism. The findings raise important safety issues while providing a potential model for examining aspects of causation and disease pathogenesis in acquired disorders of behavior and development.
Thanks KWombles!
“Compared with unexposed animals, exposed animals showed attenuation of amygdala growth”
Ah…attenuated? The amygdalas of the vaccinated (exposed) monkeys grew with time, but shrank for the unvaccinated monkeys. Looks like something happened between 2008 and now.
Ironically, I see D. Atwood from the department of chemistry, U. Kentucky in the author list. He’s the inventor of the chemical sold as OSR. I see that the U. Kentucky team isn’t on the author list any more.
Here’s a comment by Mark Blaxill and Dan Olmsted over at the age of autism:
From age 4 to 6 months, the brains of the unvaccinated animals shrunk slightly (volume went from 78185.22 to 78097.05)
During the same period, the total brain volume for the vaccinated monkeys increased notably: from 84847.44 to 87476.46
A feature of most animals is that their brains grow during infancy. Brains shrinking during infancy (as with the unvaccinated monkeys) is generally not considered a good sign.
That’s my own understated way of saying–way to spin the flaws in the study, Dan and Mark. I bet your book will be at least as good.
That is actually a bit funny.
Isn’t this exactly the same problem of the infamous first-baby-haircuts study by Holmes, Blaxill & Haley (2003)? That is, their control group was very different to a general population reference, while their cases were just like the general population.
I don’t think it’s a coincidence that anti-vaxers tend to find such unusual control groups. When you have to fiddle with the data to get the results you want, what can you expect?
It’s also interesting that anti-vaxers have been trying again and again to produce some credible monkey-study results for, what, 2 years now? It’s kind of pathetic that this is the best they can come up with. This is their absolute best shot.
In fairness, it’s very hard to get anything right when reality is not on your side.
Finally, a very simple observation: If you can detect considerable amygdala development abnormalities by looking at only 12 monkeys (vs. 4 in the control group) shouldn’t just about every vaccinated human child have an abnormal amygdala?
Great job Sullivan. Catherina and I will try and tackle this one too next week. The comments about this over on AoA are so rich; it is obvious that no one has bothered to actually read it, let alone understand it.
So not only did overall amygdala size non-significantly increase over that of their whopping control group of n=2, but they never made any claims that the observed increases were abnormal. Did I miss the part where they explained how and why they ended up with the losses in both of their groups?
I think the more apt analogy is the last undead out of the grave… all the more fitting that this turned up in eastern Europe.
I don’t doubt that this is the same research project that kicked out the Neurotoxicology paper, with most of the same animals. It’s been clear all along that Wakefield’s crew were willing to throw in new animals at any point when it served their purposes.
This is a minor journal: They report, “Its circulation varies from 300 (regular issues) to 950 (conference issues).” They also indicate relatively short “turnaround” time: “The average time from submitting a manuscript to print is 6 months. ” Also of interest: Their content is available for free online. So, unlike Neurotoxicology, they don’t appear particularly vulnerable to boycott.
Pardon me for being dense: but doesn’t this prove that not vaccinating is bad for neural growth? It looks like if the baby does not get vaccinated, then they are being hit with antigens that are removing brain mass.
Edit to add: David are you hinting that there might be a Hitler Zombie posting by Orac soon? I guess it is up to him, but it is too obvious to the point of being silly.
I prefer the term “kudlak”, but then nobody knows what I’m talking about. Semi-seriously, zombies, like most people’s notion of a “vampire”, are creatures of cinema. I prefer to discuss at what people really believed in, and “kudlak” is among the latest titles in authentic belief.
I think some kind of post from Orac is inevitable.
Excellent to read a calm and rational view of this paper.
It’s interesting that Age of Autism is actively suppressing posts that point to the fact that amygdala volume normally increases with growth in macaques, just as it did in the vaccinated group in the Hewitson study.
When Betty Watson asked: “If no one has studied non-human primate amygdala development, how do we know what ‘normal’ is? Is it normal to have a decline in amygdala size during development?”, a reply was posted which quite incorrectly claimed that “Normal is the control group that was only given injections of saline.” As Sullivan pointed out, the evidence clearly indicates that in this case that statement is false, and that Hewitson’s conclusions are flat-out wrong, but you won’t learn that in the AoA echo chamber.
brian,
I am not surprised. It is amazing that they wrote the post at all. Took a lot of chutzpah to write such a glowing review of such bad science. Once they did that they can’t back down and admit that it is junk.
I wonder if the authors missed the study cited above on the growth of the amygdala in macaques? Were they that sloppy? I found it with less than 10 minutes searching. They are supposedly primate researchers. The Hewitson paper was “Received 1 April 2010, accepted 30 June 2010”. The paper on amygdala size was published in September 2009.
Let’s assume they hadn’t seen the paper. Let’s assume they are going on general knowledge. How can they assume that a shrinkage with time of a major portion of the brain is normal?
It is mind boggling that they had the guts to put this into print. It further boggles the mind that some referee approved it. Well, maybe that isn’t so mind boggling. It is quite within the realm of reason that, say, the Geier’s refereed the Hewitson paper, Hewitson refereed the DeSoto paper, and so on….
I suppose the fact that not only Hewitson and her colleagues but also the reviewers and editors of Acta Neuorbiologiae Experimentalis missed a study that directly refuted the conclusions of the Hewitson paper says quite a lot about Hewitson and the reviewers and editors, but perhaps they just missed the most important paper related to the topic although it was published over six months before Hewitson submitted her article.
However, it’s clear that whoever controls the comments related to Olmsted and Blaxill’s post at Age of Autism, whether it is Olmsted or Blaxill or a subordinate, is not only suppressing the truth, but lying.
According to a comment by Jake Crosby at RI (in response to my confronting him about a comment which a) assumed I was Kev, b) assumed I was paid by a pharmaceutical company and c) equated my perfectly truthful description of myself as a theology student with a “front” for a con), it is Kim stagliano who does the “moderating”. I have emailed Stagliano before, and gotten automatic replies saying that, because she is very busy, said email may or may not be read. Based on this, I suspect that AoA’s draconian “moderation” is not ONLY the result of intentional censorship of dissent and corrections, but also undue power in the hands of someone who may not spend enough time on her task to be fair and thorough even if she wanted to.
A big factor in this study that would make the results questionable no matter what the sample size is that the monkeys were isolated from their mothers. Isolation of monkeys from their mothers produces autism-like symptoms all by itself.
Classic Wakefield. Be interesting to see if Laura Hewitson has the integrity to write to Age of Autism, or indeed LB/RB, to explain these points.
Plus ca change Thoughtful House.
Wow.
This so… unbelievably…
Bad science.
I think any first year science student would realize that there are many problems.
These people must have been bribed.
Now more monkeys are going to be tortured in order to prove this study was ridiculous and unfounded.
I’m wrapping up a response to this study. My major conclusion: Hewitson was only willing to claim “statistically significant” shrinking in the LEFT amygdala, which could be explained by NORMAL asymmetry in the brain. As for the horror show MRI images, I think a sufficient explanation is that they simply cross-sectioned a lower part of the unexposed brain where development was particularly lopsided.
David,
what concerns me is the big block of missing tissue at about 4-O’Clock on the MRI scan of the control subject. What is up with that?!?
I just snorted out coffee after reading that!! (Laughing of course) This is the reason I so dearly love Left Brain/Right Brain!! But now I must go clean myself up! Excellent work here!! Might I have permission to email a link to this article around to some people I know who really need to read this?
Theo,
no permission needed to email a link to any article. Feel free!
To not be messed up with another Theo ahem…Theodora (blech!!!),I wanted to tell you all Theo stands for Theophano, tis a nickname of mine. I’ve been mixed up elsewhere.
Thanks for following up on this, Sullivan.
I don’t understand why Mark Blaxill and Lyn Redwood are promoting this failed study, since the sloppy work, its promotion by SafeMinds and Age of Autism, and the vigorous suppression of the truth at Age of Autism provides additional conclusive proof to those who accuse SafeMinds and Age of Autism of ignorance, bias, and of being anti-science. That’s just wacky.
Even if Hewitson et al. hadn’t shot themselves in the foot (or head) by using a ridiculously tiny control group, it happens that these studies are actually technically difficult, as “obtaining volumetric measures in infants is … difficult, as structures are less well defined and largely unmyelinated, making segmentation challenging. Moreover, it is still unclear whether individual anatomic variation across development, in healthy, normally developing infants, is reflected in the configuration and function of the mature brain.” [“Neuroimage. 2010 Feb 1;49(3):2791-9. Epub 2009 Oct 19]
Of course, it’s actually a bit odd to focus on amygdala development in macaques who, like these animals, were removed from their mothers as neonates, since that in and of itself affects the amygdala.
This is just as comical as SafeMind’s support for the laughable Uhlmann (and Wakefield) study despite what may be the most thorough takedown of a paper in the history of molecular biology. I don’t know why Blaxill chose to have “LOSER” tatooed across his forehead, but it is remarkable that he can’t read the the word when he looks in the mirror–everything’s backwards for him, I suppose.
Sullivan,
“what concerns me is the big block of missing tissue at about 4-O’Clock on the MRI scan of the control subject.”
I was struck by that too. It immediately made me think of a hematoma, though I don’t know if it would look like this. After looking into it a little further, I think it’s simply a space between the hind brain and a low-hanging piece of the frontal lobes. Something it helped me to remember: These MRI scans are, effectively, two-dimensional representations of a three-dimensional object, and as such all kinds of weirdness is possible merely from misleading appearances.
I hope to get the essay up in the next 24 hours or so.
Here’s a link for my article on this:
Click to access 11monkeys.pdf
Something I learned just after I posted this: Apparently there are at least some reports that shrinking of the amygdala, among other parts of the brain, is reported in AUTISTIC children. So, if Hewitson’s observations are accurate, and reveal anything of potential interest related to autism, it is that at least one UNvaccinated monkey developed autism-like features.
Hi David N. Brown –
Indeed, there appear to be studies showing both enlarged, or decreased amygdalas in autism. To my mind, it isn’t clear if this is a problem with the studies, or that just having any thing other than ‘just right’ be a problem. In either case, however, it tends to make this study look pretty ridiculous.
– pD
You said, “half of the control group are missing in the data? What’s the reason for that? No, that’s a real question which I can’t find answered in the paper: what happened to the two other controls?” The article actually did address this in stating, “one control had to be withdrawn due to a scheduling an error.” I don’t know about the other one but would like to find out as well.
Having a conflict of interest doesn’t invalidate the study. There were 4 other authors involved in the study yet you completely focus on Dr. Hewitson as if discrediting her is going to discredit the entire study. This shows an obvious bias.
The study in question does attempt to explain the decrease in brain functioning (and therefore volume):
“Maturational changes seen in the experimental setting may be relevant to
the observed decline in [11C]DPN-binding in the unexposed animals over time. In rats, [MET5]-enkephalin, ?-endorphin, and opioid-receptor levels in the cerebellum reached their highest level in the first few weeks post-partum, and subsequently declined to low levels (Tsang et al. 1982, Zagon and McLaughlin 2004). Others have demonstrated a significant 2- to 4-fold
decrease in the concentration of opiate receptors in human fetal brain tissue during the last trimester of pregnancy (Kinney et al. 1990). This may be attributable to a normal process of programmed neuronal apoptosis that can extend into infancy, and is consistent with the enlarged amygdala volume and [11C]DPN-binding in exposed animals over time that was observed here.”
The study is listed as a pilot project and indicates a causal relationship between the vaccine schedule and autism spectrum disorders. If people were really interested, they would follow up on this study:
“the results of this pilot study warrant additional research into the potential impact of an interaction between the MMR and thimerosal-containing vaccines on brain structure and function.”
The authors address the small sample size here: “While, as a pilot study, the size of the study groups limits the strength of the conclusions that can be drawn, the use of statistical modeling and repeated measures contributed to the study’s power and increased the accuracy of the estimates.” and here:
“We purposefully assigned a larger number of animals to the exposed group in order to optimize the chances of observing what we anticipated to be an uncommon or idiosyncratic effect.”
Although it’s a just a pilot study, the fact that there was a statistically significant connection between the exposed group and autism shouldn’t be overlooked.
Brian,
you appear to be doing a cut and paste from a blog post which linked to mine.
“Having a conflict of interest doesn’t invalidate the study.”
Please educate those who throw out any study funded by or performed by the CDC or drug companies.
“Although it’s a just a pilot study, the fact that there was a statistically significant connection between the exposed group and autism shouldn’t be overlooked.”
First–statistically significant and connection are different things. A statistically significant correlation could suggest a connection but it is not a connection.
Second–I don’t think the data above are good enough to claim a statistically significant result. Based on the data that are out on amygdala sizes in growing macaques, the noise is too high. There are a lot of scatter in the measurements and the authors above are not taking that into account.
“We purposefully assigned a larger number of animals to the exposed group in order to optimize the chances of observing what we anticipated to be an uncommon or idiosyncratic effect.”
Too bad for them. They purposely made it difficult to get statistical significance. Much more to the point, too bad for the laboratory animals whose lives were wasted.
“The study is listed as a pilot project and indicates a causal relationship between the vaccine schedule and autism spectrum disorders. If people were really interested, they would follow up on this study:”
If the people promoting Ms. Hewitson’s research really cared, they’d know that Gene Sacket *is* doing a followup on the study.
The study does not attempt to explain anything. The authors “attempt” to do a great deal. They are, unfortunately, limited by the very poor design and execution of this study.
“Having a conflict of interest doesn’t invalidate the study. There were 4 other authors involved in the study yet you completely focus on Dr. Hewitson as if discrediting her is going to discredit the entire study.”
Factually incorrect. I focus on the poor study. Ironically, you are doing excatly what you blame me of–attacking the person rather than the message.
“I don’t know about the other one but would like to find out as well”
I guess you haven’t read Ms. Hewitson’s response to the critique Steven Novella submitted to the journal?