[Neurosciences/Junk Sciences] Autopsy of a flawed study of aluminum and brain inflammation (Li et al., J Inorg Biochem 2017)

Note: This is a special blog post coauthored by The Mad Virologist and The Blood-Brain Barrier Scientist (this article will be co-published on both our blogs). Another post has already been published on this paper, but we wanted to take a deeper look at everything that is wrong with this paper.

[UPDATE2] The study in question got retracted according to RetractionWatch:
http://retractionwatch.com/2017/10/09/journal-retract-paper-called-anti-vaccine-pseudoscience/

[UPDATE] I would strongly recommend the reader to look at the comments on Pubpeer about this paper. It is terrifying to think how it percolated through peer-review.
https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983#undefined

A recent paper by ophthalmologist Chris Shaw was published and immediately touted as being proof positive that the aluminum adjuvants found in some vaccines are responsible for causing autism. Before we get into the paper, I have a few choice things to say about Chris Shaw. Despite not being an immunologist, Shaw has ventured into studying how vaccines and vaccine adjuvants cause neurological disorders such as autism. Shaw made headlines in 2016 when a paper he co-authored that claimed to show a link between the HPV vaccine and neurological disorders was retracted after being accepted by the journal Vaccine. It turns out that the statistics used in the paper were completely inappropriate and there were undisclosed conflicts of interests for some of the authors, including Shaw.These issues should have prevented the paper from being accepted in the first place, but mistakes do happen and science tends  to be self correcting. More surprising is that Shaw claimed that he didn’t know why the paper was retracted and that the science was of the highest quality. Shaw’s previous work has also been described by the WHO as deeply flawed and rejected by that body. This isn’t being brought up to dismiss the paper out of hand but to help illustrate why Shaw’s work is deserving of additional scrutiny. Hopefully by the end of this post, the logic behind the need for additional scrutiny of anything Shaw publishes is abundantly clear. We’ll begin by examining the methods used by Shaw’s research group and point out some of the issues.

Background for experimental design flaws: PK and species issues

One problem that is recurrent with Shaw is his “vaccination schedule” tries to consider rodents, such as mice and rats, as humans in miniature. It is wrong to assume that rodent and human primate species are alike, they’re not and there are notable physiological differences between rodents and non-rodents. For example, there are a couple of studies by Terasaki and colleagues (http://onlinelibrary.wiley.com/doi/10.1111/j.1471-4159.2011.07208.x/abstract) that have shown differences in the expression of solute carriers and drug transporters at the blood-brain barrier. We cannot exclude that such differences may bias the outcome observed in his studies, but this bias applies intrinsically to any in vivo studies based on a rodent model.
There is also the issue of brain development and mapping the vaccination schedule and the brain maturation. In this study (as well in the previous ones), Shaw and colleagues consider that applying vaccines from post-natal day (PND) 3 to 12 is representative of a human infant vaccine schedule. There is some differences in the literature, as previous studies from Clancy and colleagues mapped the PND12 to the 7th gestational months in humans (https://blogs.cornell.edu/bfinlay/files/2015/06/ClancyNeurosci01-17kkli7.pdf), some more recent publications map PND21 to 6th month post natal in humans, making the PND12 around the 3rd month infancy following full-term birth (http://www.sciencedirect.com/science/article/pii/S2352154615001096). You can easily appreciate that by following Shaw flawed experimental design, the total amount of Al administered during a 2 year period has been indeed administered within 90 days of birth, whereas the vaccination schedule according to the CDC does not start before the 2nd month of infancy if we exclude the two injections of Hepatitis B vaccines at birth and after the first month respectively (https://www.cdc.gov/vaccines/schedules/hcp/imz/child-adolescent.html).

In addition to a flaw in the experimental design, we cannot exclude some differences in the pharmacokinetic profile of Al adjuvants between mice and humans. The data available is fairly limited but a recent study from Kim and colleagues (https://www.ncbi.nlm.nih.gov/pubmed/26437923) failed to show a significant brain uptake of Al compared to controls following the single oral administration of different Al oxide nanoparticles at a concentration of 10mg/kg. Furthermore, the approximation of Shaw in terms of total burden of Al from vaccines (550 microg/kg) is not an accurate metric as we have a dynamic process involving absorption, distribution and elimination to occur simultaneously. A daily burden of Al from vaccines is a much more reliable parameter to consider. Yokel and McNamara (https://www.ncbi.nlm.nih.gov/pubmed/11322172) established it about 1.4-8 microg/day for based on 20 injections spanning over a 6-year period in a 20kgs individual.
If we consider Shaw calculation, then the total burden at age 6 would be 1650 microg/kg or 33’000 microg for a 20kgs 6-year old child. That’s about 15 microg/day of daily Al burden from vaccines, a value that is 2 to 10 folds higher than applied to humans. It makes therefore very difficult to compare apples to oranges, as Shaw experimental paradigm is flawed and not representative of a clinical scenario.

Selection of genes to measure:

Selecting which genes to measure is a crucial step in a study like this. If care is not given to ensure that the correct genes are selected, then the study will be a wasted effort. Shaw stated in the paper that they selected genes that were previously published. However, not all of the genes that they measured came from this paper. Only 14 of the genes were from this paper (KLK1, NFKBIB, NFKBIE, SFTPB, C2, CCL2, CEBPB, IFNG, LTB, MMP9, TNFα, SELE, SERPINE1, and STAT4). This leaves 17 genes the were measured but not found in the paper. Two of these can be explained. One gene, ACHE, was mentioned as having been selected because of other work, so it is sourced. The second gene, is the internal control gene beta-actin. This is a housekeeping gene that is often used as an internal control to provide a relative expression from. This leaves 15 genes unaccounted for. We suspect that these genes were selected because they are involved in the innate immune response, but no reason is stated in the paper.

The way these genes were selected is problematic. Because half of the genes seemed to be selected for uncited reasons, this study is what is known in science as a “fishing expedition.” There’s nothing inherently wrong with this type of research and indeed it can lead to new discoveries that expand our understanding of the natural world (this study that increased the number of sequenced viral genomes by nearly tenfold is a good example of this). But what fishing expeditions can show is limited. These types of studies can lead to other studies but they do not show causality. Shaw is claiming causality with his fishing expedition here.

There is also the problem that they used old literature to select their gene targets when much more recent research has been done. By happenstance, they did measure some of these same genes in their study. However, their results do not match has has been measured in children that have been diagnosed with autism. For example, RANTES was shown to be decreased in children with autism. In Shaw’s work there was no statistical difference in RANTES expression between mice given the aluminum treatment and those receiving saline. Likewise, MIP1alpha  was shown to be decreased in developmentally delayed children but was shown to be increased in the aluminum treated mice. This was also the case for ILIb which was found to be elevated in children with moderate autism yet there was no statistical difference between the mice receiving the aluminum treatment and those receiving saline. In fact IL-4 was the only gene to follow an expression pattern similar to what was found in children with severe autism (elevated in both cases). However, there is something odd with the gel in this case. This was the image for figure 4 that was included in the online version of the paper (we have not altered the image in any way). Look closely at the top right panel at the IL-4 samples and the IL-6 samples. You’ll notice that the bands for the control and the aluminum treated mice have different color backgrounds (We enlarged the image to help highlight this but did not adjust the contrast). If these came from the same gel, there would not be a shift in color like this where the treated bands have a lighter color encircling them. The only way this could happen is if the gel was assembled in photoshop. The differences could be real; however, since this image was modified we do not know for sure and this is scientific misconduct. Papers get retracted for this all the time and people have lost their degrees for doing this in their dissertations. These gel results cannot be trusted and the paper hinges on them. The Western blots and issues with them will be discussed below.

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The unaltered figure 4.

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A close up of the panel with the regions in question highlighted.

Semi-quantitative RT-PCR:

In order to quantify the gene expression levels of the genes that Shaw’s group selected, they used an older technique called semi-quantitative RT-PCR. This technique uses the exponential increase in PCR products in order to show differences between expression of a gene under different conditions. There’s nothing wrong with the technique provided one understands what the limitations are. Let’s say you have a large number of genes that you want to measure expression of, but you aren’t sure which genes are going to be responsive and you have limited funds. Semi-quantitative RT-PCR is a good method to screen for specific genes to be examined further by more precise techniques, such as Real-Time RT-PCR, but it’s not appropriate to use this technique and then make statements about precise quantification. Where semi-quantitative RT-PCR excels is with genes that are normally not expressed but can be expressed after some sort of stimulus, such as terpene biosynthesis genes that are induced by insect feeding.

To put it bluntly, semi-quantitative RT-PCR was not used properly in the paper by Shaw. The way that it was used implied that it would be quantitative when the technique is not that precise. Without verification by another method, ideally Real-Time PCR which can determine what the exact abundance of a given target is, these results should be taken with a grain of salt. This would still be the case if there weren’t irregularities in the gel images. With those irregularities, this is absolutely essential and should have prevented this paper from being accepted.

Western-blots and data manipulationPCR and Western-blots data: the owl is not what it seems
As The Mad Virologist mentioned, the semi-quantitative PCR is an old-fashioned RNA quantitation method, with the use of Real-Time quantitative PCR (that quantifies the amplification product at each cycle, using a fluorescent dye as an indicator) is a much more accepted method nowadays (see his section for more details). For Western-blots, the semi-quantitative approach is more accepted but it is important to show data that are consistent between what you show (qualitative) from what you count (quantitative). In Western-blot analysis, we measure the relative darkness of a protein band (the black lines that you see in papers) between treatments and controls. Because you cannot exclude some errors due to the amount of protein loading, we also measure the band intensity for proteins that are very abundant, usually referred as housekeeping proteins (because they play essential functions in cells). In this case, beta-actin (named ACT in the paper was used).
Once you normalize to beta-actin, you can compare the effect of a treatment by comparing the relative band intensity ratios. In both cases (semi-quantitative PCR and Western-blots), “what you see is what you measure” or you have to show a “representative Western-blot” alongside a quantitative data to demonstrate that your quantification matches with band densities. The common practice is the use of image acquisition software like ImageJ to determine band density. Showing Western-blot is nice, but not foolproof. Indeed, Western-blots data (with fluorescence images) is amongst the most common method by which some researchers can manipulate or even falsify data but also the most common type of data that spark a paper retraction. Someone notice something fuzzy on a Western-blot data, creating some questioning reaching to the editors and asking access to the full dataset (usually the X-ray film or the original full scan of the blot). Often, the author will use the excuse “the dog ate the flash drive” or “the hard drive containing the data crashed” if they cannot provide such data.
There are some methods to spot some image manipulation on Western-Blots and include playing with the brightness/contrast, requesting the presence of quantitative data in addition of a representative blot, samples must be coming from a same gel (you cannot use a cookie-cutter and build-your-own perfect gel). There is an excellent article that describe the pitfalls and cases of bad Western-blot data representation if not image manipulation. (https://www.elsevier.com/editors-update/story/publishing-ethics/the-art-of-detecting-data-and-image-manipulationThere are, at this time, different issues raised both in the Western-blots pictures and their subsequent analysis raising the reliability of the data presented in this study.

In this post, we have used the full-resolution pictures provided by the journal website (http://www.sciencedirect.com/science/article/pii/S0162013417300417), opened just pictures in ImageJ to convert such pictures into 8-bit format, invert the lookup tables (LUT) and adjusted the brightness and contrast. We have exported such pictures in Powerpoint to ease the annotation and comments. We recommend the reader to judge by himself/herself and download the full-resolution images as well.

The first concern is by looking at Figure 1C. First, this is the original Fig.1.

1-s2.0-S0162013417300417-gr1_lrg

Then, this is the close-up analysis for Fig.1C

Slide1

There are several issues. First there are some bands that appears as band splicings, in which the author create a custom blots by assembling different bands from different gels. This is a no-no in Western-blots: all bands showed in a blot should come from the same gel. This is why Western-blot is a torture for graduates students and postdocs, you need to show your best blot with all bands showing the same behavior for your quantitative analysis.
Second, the presence of a rectangular grey piece that was added on the top of control 3 TNF band. This is a possible data manipulation and fraud, as you are voluntary masking a band and hiding it. Thats a big red flag on the paper. The third issue of Fig.1C is the consistent feeling of seeing bands either cropped on a grey rectangle or what I call a “Photoshop brushing” in which you brush off using the brush function area of the gel you consider not looking good enough. You can clearly see it with actin as we have a clear line between the blurred blot and a sharp and uniform grey in the bottom half of the blot, compared to the wavy top of the blot. This a grey area that I am not familiar with Western-blot but this is a no-no for any immunofluorescence picture. Any image manipulation that goes beyond the brightness/contrast adjustment and involves alteration of the acquired picture is considered as data manipulation. If you analyze the data upon correcting for the inconsistency of Figure 1C, the graph looks much more different and failed to show any differences between Al-treated and control, when you restrict yourself in over-normalizing it and plot straight the protein/actin band density ratios.

What is also concerning and surprising is the conclusion from the authors that males, not females, showing an inflammatory response. Of course, the authors failed to show the same outcomes from female animals and expect us to trust them on this. The problem is that such conclusion is in direct contradiction with the literature. There is a solid literature supporting the presence of a sexual dimorphism in terms of inflammatory response, in particular in terms of neuroinflammation and autoimmune disorders such as multiple sclerosis (https://www.ncbi.nlm.nih.gov/pubmed/28647490; https://www.ncbi.nlm.nih.gov/pubmed/27870415). There is also a growing call to the scientific community to provide results for both sexes (males and females alike). Although Shaw reports the study was performed in both males and females, he gives us this explanation at the end of section 3.1: Taken together, a number of changes indicative of the activation of the immune-mediated NF-κB pathway were observed in both male and female mice brains as a result of Al-injection, although females seemed to be less susceptible than males as fewer genes were found altered in female brains.

Yet the interesting part comes when Shaw try to compare ikB phosphorylation between males and females following Al injection (Fig.3C). When you analyze the data, you are raising concerns very rapidly. First, we have a possible case of cookie-cutter band in which you just paste a band that seems nice enough in a blank space. This is a very suspicious activity as you can make up data as easy as this. Second, there is again this “Photoshopping brushing/erasing” taking place in that figure, in which I suspect a case of fraudulent activity. As you can see in female, it is as if someone tried to mask some bands that should not have been here. Remember when he said that males but not females showed an inflammatory response? Is it trying to dissimulate data that contradict his claims?

image

Again, lets bring up Figure 3 at its full resolution.
1-s2.0-S0162013417300417-gr3_lrg

Finally, the same issues are persistent and even more obvious in Fig.5A. Again, we have a mixture of different Western-blots image manipulations including bands splicing, Photoshop brushing, cookie-cutter bands……

First, the unedited picture:
1-s2.0-S0162013417300417-gr5_lrg

And below the close up of Fig.5A

Slide3

These are some serious concerns that raise the credbility of this study and can only be addressed by providing a full-resolution (300dpi) of the original blots (X-ray films or the original picture file generated by the gel acquisition camera).  There has been a lot of chatter on PubPeer discussing this paper and many duplicated bands and other irregularities have been identified by the users there. If anyone is unsure of how accurate the results are, we strongly suggest looking at what has been identified on PubPeer as it suggests that the results are not entirely accurate and until the original gels and Western blots have been provided, it looks like the results were manufactured in Photoshop.

 

Statistics:
Long time followers know that I tend to go right to the statistics that are used in papers to see if what they are claiming is reasonable or not. Poor use of statistics has been the downfall of many scientists, even if they are making honest mistakes. It’s a common problem that scientists have to be wary of. One easy solution is to consult with a statistician before submitting a paper for publication. These experts can help point out if the statistical tests that were run are the correct or not. The Shaw paper could have benefited from this expertise. They used a Student’s T Test for all of their statistics comparing the control to the aluminum treated. This is problematic for a couple of reasons. These aren’t independent tests and the data likely does not have a normal distribution, so a T Test isn’t appropriate. Better statistical tests would have been either Hotelling’s T-squared distribution or Tukey’s HSD.  Another issue is how the authors used standard error (SE) instead of standard deviation (SD). To understand why this matters, it helps to understand what the SE and what the SD measure and what these statistics show. The SD measures the variation in samples and how far the measurements are from the mean of the measurements. A smaller SD means that there is low variability in the measurements. The SE measures the likelihood that a measurement varies from the mean of the measurements within a population. Both the SE and SD can be used; however, using the SE is not always appropriate, especially if you are trying to use it as a descriptive statistic (in other words if you are trying to summarize data). Simply put, the SE is an estimation and only shows the variation between the sample mean and the population mean. If you are trying to show descriptive statistics, then you need to use the SD. The misuse of SE when the SD needs to be shown is a common mistake in many research publications. In fact, this is what the GraphPad manual has to say about when to use the SD and when to use the SE:

If you want to create persuasive propaganda:
If your goal is to emphasize small and unimportant differences in your data, show your error bars as SEM,  and hope that your readers think they are SD. If our goal is to cover-up large differences, show the error bars as the standard deviations for the groups, and hope that your readers think they are a standard errors.” This approach was advocated by Steve Simon in his excellent weblog. Of course he meant it as a joke. If you don’t understand the joke, review  the differences between SD and SEM.” The bottom line is that there is an appropriate time to use the SE but not when you are trying to summarize data.

Another issue is the number of animals used in the study. A consensus in published study is to provide a minimal number of animals (usually n=8) needed to achieve statistical significance but also maintain to a minimum to ensure proper welfare and humane consideration for lab animals. In this study, such number is half (n=5). Also the authors are bringing some confusion by blurring the lines between biological replicates (n=5) and technical replicates (n=3). By definition, biological replicates are different organisms that are measured and are essential for statistical analysis as these replicates are independent from each other. Technical replicates are dependent on each other as they come from the same biological samples and are repeated measurements. By considering the latter as statistical relevant, you are biasing yourself to consider a fluke as a biological phenomenon.

 

Conclusions:
Based on the methods that were used in this paper, Shaw et al. went too far in declaring that aluminum adjuvants cause autism. But there are six other key points that limit what conclusions can be drawn from this paper:
1) They selected genes based on old literature and ignored newer publications.
2) The method for PCR quantification is imprecise and cannot be used as an absolute quantification of expression of the selected genes.
3) They used inappropriate statistical tests that are more prone to giving significant results which is possibly why they were selected.
4) Their dosing regime for the mice makes assumptions on the development of mice that are not correct.
5) They gave the mice far more aluminum sooner than the vaccine schedule exposes children to.
6) There are irregularities in both the semi-quantitative RT-PCR and Western blot data that strongly suggests that these images were fabricated. This is probably the most damning thing about the paper. If the data were manipulated and images fabricated, then the paper needs to be retracted and UBC needs to do an investigation into research misconduct by the Shaw lab.

Maybe there’s a benign explanation for the irregularities that we’ve observed, but until these concerns are addressed this paper cannot be trusted.

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[Sciences/Junk Sciences] Remember the deadly turmeric IV infusion done in a holistic clinic? Lessons from the FDA report

You have remember this story of this young woman that died shortly after recieving an IV infusion of turmeric acid (aka curcumin, a bioactive compound found in Curcuma) in a holistic clinic in California few months ago (http://www.10news.com/news/team-10/encinitas-woman-dead-after-i-v-infusion-of-turmeric).
This story baffled me for many reasons. First, it was really puzzling me on how quack medicine (rebranding itself as “holistic” and “integrative” to appear more sciencey) have been moving slowly but surely into medical procedures normally held by medical staff, with some dubious claims of “IV therapy” in which the onset of an IV line and pumping up vitamins straight into your systemic circulation will help you “detox” or “rejunevate”.
Second, how turmeric acid that have been bounced by some “health/food gurus” as superfood (move on kale and quinoa, you are so 2015!) quickly moved on as therapeutics without even having the right science to back it up (until now only preclinical studies done in cells grown on Petri dishes and in rodents), with the glittery “cures-it-all” sticker all over it.
You see, turmeric acid is way far from being the next wonder drug as sold by woo peddlers. Why? Lets see some of its features (https://pubchem.ncbi.nlm.nih.gov/compound/curcumin#section=Top).
First thing, turmeric acid has a problem. A huge problem. This problem is solubility. It has a calculated xLogP of 3.2, this is already telling us this compounds is lipophilic (likes fat). It will dissolve well in oil, but not well in water (less than 0.1mg/mL according to Santa Cruz Biotechnology data sheet). If you try to go beyond that value, you will have a saturated solution with turmeric acid precipitates. These precipitates can have serious effect if injected into an IV line, if these particles are big enough to clog some capillaries.
You can circumvent things around by tweaking nanoparticles carriers. Still, even from food intake, turmeric has a very low bioavailability. From 100g of pure turmeric acid swallowed, only 1g will effectively reach the circulation and circulate through your body.
The second problem with turmeric is its pharmacokinetic profile. According to the reference cited by the FDA report, turmeric is highly unstable at physiological pH (7.4). According to this review, the elimination half-life (t1/2) for turmeric is very low (1.7+/-0.5h). By 6 hours, most of the turmeric injected via IV route will be gone. Therefore, if turmeric was considering for therapeutic, it would require multiple dosing that are either ridiculous (Dosing interval of about ~2 hours, therefore swallowing a pill every two hours) or being on a constant IV infusion (that is not realistic for everyday life).
Third problem with turmeric? Its pharmacological activity. Two important parameters have to be accounted for a drug candidate: its selectivity (does the drug targets one or several proteins?) and IC50 (what is the concentration needed to achieve 50% inhibition).
The problem with turmeric is that it is considered as a “dirty” molecule because it hits a bit of everything, with many signaling pathways affected by it. The second problem is its very high IC50. Anti-cancerous activity of turmeric swings around 10microM in various cancer cell lines in a Petri dish and have other targets at higher doses. This is not a horrible value, not a good value either. Usually we want to reach an IC50 in the nanoM range (10’000 less concentration than 10µM). Thats not the case for turmeric. Maybe by tweaking the chemical structure we may improve its IC50, but since the compound itself has so many targets trying to optimize it for therapeutic purposes maybe simply a waste of time.
If we stick to the 10µM concentration and an average molecular weight of 328g.mol-1 for turmeric, we need a concentration of 3mg/L or (0.003mg/mL) to expect some biological activity. Now the problems come in with the FDA report. There are two reported cases of adverse events, including the fatal cases. In both cases, patient had an IV line of turmeric acid. In both cases, both patients were mentioned an IV infusion of turmeric acid at 10mg/mL. First, this concentration would have made no sense. It is 300 times higher than the hypothetical dose needed to achieve a biological activity in vivo. Second, the final concentration in the IV bag was much less than this concentration, as the FDA reported only 1% of the prescribed concentration was found in the IV bag (0.00235mg/mL).
Someone has been not only been deceiving their customers by selling you less product than advertised (1% net content is honestly a huge rip-off) but also had absolutely no clues on what they were injecting. So we can blame two actors: either the compounding company that prepared the turmeric or the holistic clinic (I guess you can point who is the crook in the story).
Both cases involved ImprimisRX, a compounding pharmacy. These are laboratories under the responsibility of a pharmacist holding a specialization in compounding. He or she has to follow established rules and protocols, adhere to good manufacturing procedures in compliance with the FDA. It seems there is no wrongdoing from the compounding. The compounding produced an emulsified form of turmeric (to increase its solubility). Yet, the final concentration in the vial was about 0.205mg/mL or about 2% of the amount put on the label. Since turmeric is highly unstable under aqueous solution (even in its emulsified form) we cannot exclude a degradation of the product from the time it got compounded to the time it was administered. In aqueous protein-free solution, 90% of turmeric acid is degraded within 30 minutes (https://www.fda.gov/downloads/AdvisoryCommittees/CommitteesMeetingMaterials/Drugs/PharmacyCompoundingAdvisoryCommittee/UCM466380.pdf).
Now comes an another problem: was there any deception between ImprimisRX and the holistic clinic? One of the reason is the use of polyethylene glycol 40 (PEG40) castor oil in the compounding process. PEG40 may contain traces of diethylene glycol (DEG), a very well known toxic compound if ingested, with a toxicity of about 1mg/kg. DEG is a byproduct of PEG production, therefore the FDA has different quality grades of PEG whether it is destined for non-medical usage or for human consumption. DEG was found at a concentration of 0.21% (0.21g pure DEG in 100g of PEG40). The PEG40 used for the compounding was 1.25% with the clear label “CAUTION: For manufacturing or laboratory use only.” Why the compounding pharmacy used that ungraded PEG40? We don’t know yet.
PEG40 oils are used for cosmetics and considered safe for cosmetics usage  because the bioavailability of this compound is small and suited for topical application (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4505343/). But you have to remember that a product that is considered safe in an administration mode is not in another administration route. This is probably explaining the adverse effect observed.
The second possible issues is a possible allergic reaction to PEG40, as the FDA cited previous reports of allergic reactions of patients exposed to PEG formulations following IV infusion of anti-cancerous agents.
By now, your enthusiasm for cur turmeric should (rightly) wind down. Turmeric is certainly great for spicing your food, not much for your health. But most of all, don’t let a “holistic clinic” perform any type of IV infusion.
IV infusion is a very delicate procedure, used for restricted applications (chemotherapy, anesthesia, infectious disease……) by a trained personnel in a medical environment (hospital or medical clinic).

[Sciences/Junk Sciences] How the recent AHA recommendation on coconut oil is making many getting nuts (and why coconut oil is not an healthy choice)!

Coconut oil. Coconut oil. Yep, that same coconut oil that (almost) nobody knew about a couple of years ago and suddenly became the next big thing in fad diets. Some claimed it is healthier than vegetable oil (http://pilatesnutritionist.com/why-coconut-oil-is-better-than-vegetable-oil/; which turned out is not true), other claimed it can help you loose weight (https://authoritynutrition.com/coconut-oil-and-weight-loss/; that is also hard to imagine how to loose fat by keeping an high-fat diet) or even use it as a natural sunscreen (http://thecoconutmama.com/coconut-oil-sunscreen/; which of course will more likely help you roast like a rotisserie chicken).
You see the fad went a bit crazy with the habitual “wellness” bloggers making miraculous claim. The fact is coconut oil is no better than any oil and indeed maybe as bad as any saturated fats.
The only thing that I would say coconut oil is good, is giving you some tasty and crunchy fries that are not too greasy. Any French household know the “Vegetaline” brand (basically solid coconut oil that you mix with half sunflower oil to get a frying oil).

What is (in terms of chemical composition) coconut oil?

Coconut oil is extracted from the inner side of the coconut. It is also called copra oil. Some coconut oil are referred as “organic coconut oil” and even some referring as GMO-free coconut oil (you know the GMO-free project sticker that have no sense except operating as a form of racketeering? There are been never any GM-coconuts that hit the market. http://www.zebraorganics.com/organic-virgin-raw-coconut-oil-1-gallon-tub-zebra-organics.html?gclid=Cj0KEQjwyZjKBRDu–WG9ayT_ZEBEiQApZBFuK3KbEfSPhyNyx9z9eNUIwAmd6OwcxTWJUYKADA_fhEaAnvd8P8HAQ). Therefore, we consider all coconut oil equals (maybe slight variations between cultivars but this should not affect much the overall composition to be considered significant).

Before we discuss about the composition of coconut oil, it is important to know what a fatty acid is. Fatty acids (FA) are hydrocarbon chains (made of carbons and hydrogens) that are very similar to molecules belonging to alkanes (these are the molecules such as propane, butane and octane that are present in your propane gas tank right now fueling your grill, fueling your gas stove or fueling your SUV).
In contrast to alkanes, FA have a carboxyl (-COOH) “head” denominated and seen below:

We have two type of FA: saturated FAs (fully loaded with hydrogens) and unsaturated FAs (that have one or several C=C double bounds). Saturated FAs are usually found in fat products from animal origin (lard, butter, ghee…) whereas unsaturated FAs are usually found in plants (olive, rapseed/canola, corn, sunflower…) and in fish and seafood (usually polyunsaturated fatty acids or PUFAs aka omega- fatty acids). Unsaturated FAs either show a cis-form (like the oleic acid depicted, in which the two carbon branches are in the same side) or a trans-form (in which the two pieces of the carbon branches are opposing each other). Trans unsaturated FAs (aka trans-fats) have been already a bad rep because of their detrimental effects on the cardiovascular system (they are suspected to increase LDL levels which are known to contribute in the atherosclerotic plaques formation). Saturated FAs are also having a bad rep because they are also associated with increased risk of cardiovascular diseases, whereas unsaturated FAs (commonly found in “the Mediterranean diet”) are considered healthier.
FAs composition are usually denominated as the following: Cn:m with n referring to the number of carbons (usually an even number), m referring to the number of C=C. In our cases, stearic and oleic acid share the same number of carbon (C18) but the former has no C=C bounds (C18:0) and the latter has a C=C bound (C18:1).
Based on this table, you can see how coconut oil fares to other oils (https://www.chempro.in/fattyacid.htm)
It contains 90% of saturated FAs and 10% unsaturated FAs, whereas most of other oils commonly used in Western countries have at least 50% or more of unsaturated FAs. To give you an idea lard, tallow (beef) and butter contains 40%, 37% and 41% respectively.  You can see how coconut oil is exploding the chart.

But, but this is coming from one study and science has been wrong all the time

If you stick to mainstream media, you will get this impression right. News outlets like to sell single studies as sold and irrefutable evidence and often oversell the claims of that study. Science is never settled, especially on a single study. Many things can go wrong that result in bias. Sometimes, scientists even cut the corners and publish fraudulent data to support their claims (thats what you see a lot with anti-vaccines, anti-GMO papers, climate-deniers, creationism……).
Science build a consensus on the amount of publications and their robustness in their experimental design. When you have an overwhelming majority of papers show you a same trend, arrive to same conclusion on a phenomenon using different approaches and different observations by different groups, you reach a conclusion and set a consensus.
A consensus is only broken once you have new studies that refute the existing claims with more robust and more precise data than the existing literature. This happens very rarely as you have to being in a weight of evidence bigger than the existing literature.

The science on FAs and their effect on cardiovascular diseases is not new, this have been known for over 50 years and keep refining. This consensus built on the detrimental effects of high-fat diet is well-known and served to establish guidelines and public health recommendations. The American Heart Association, the leading association worldwide gathering both basic and clinical scientists as well as any healthcare actors establish guidelines.

The AHA has a clear statement, visible here:
http://news.heart.org/advisory-replacing-saturated-fat-with-healthier-fat-could-lower-cardiovascular-risks/
Replacing saturated fats may help to reduce your risk of cardiovascular events, in addition to an healthy (balanced) diet and physical activity.

The study that made the uproar is available here and comes from the scientific board of the AHA. You can download it for free and you can see another fat composition of different oils:
http://circ.ahajournals.org/content/early/2017/06/15/CIR.0000000000000510

As you can see, coconut oil tops the list of saturated oils and fats, followed by butter and lard. Saturated fats consumption are clearly associated with increased risk of coronary heart diseases (CHD, aka heart attack), replacement with unsaturated fats reduce such risks. Replacement with PUFAs appears even more beneficial. Such effects is not limited to CHDs, but appears involved in other diseases as well (see Figure 4).

In conclusion, dont ditch your coconut oil yet. As small amount, coconut oil is fine. What is not fine was the fad diet that was basically pushing you to switch everything to coconut oil. In my personal opinion, I would say that butter (real unsalted butter like the French “President”, Irish “Kerrygold” or Danish “Lupak” butters; not the things called margarines that were at the basis of the trans-fat problem),  was even a better alternative  than coconut oil.

In conclusion, keep your peanut oil for your deep-frying cooking, keep your canola oil for your dressings and use olive oil for cooking instead of lard and coconut oil. If the taste of coconut oil is good, just add the minimal amount needed to taste.

 

[Sciences/Junk Sciences] Essential Oils: The good, the bad and the ugly science behind some claims

Essential Oils (EOs for simplicity). These little bottles are almost everywhere. Advertised as “natural”, “pure”. Some even are trying to sell them as the next big fad. as the next “miracle cures all” remedy.

Everybody swears by EOs, giving them some curative properties despite the lack of evidence backing such claims. Their therapeutic activity is far from being demonstrated, but their ability to siphon wallets and fill bank accounts of those selling them is as efficient as  the Bernouilli’s principle.

The problem with EOs is to sort the good, the bad and the ugly science behind them.

@MommyPhD recently pointed this out in a nice chart taken from a company making a living on EO.

What I can tell, as the pharmacologist that I am, I was not only perplexed but mind-blown by this chart. It was not a nice mind-blowing effect. It was more like a trigger that turned into a “ballistic mode”.

Just see by yourself the chart below:

17634361_1898070473737773_8852685320786733500_n

What is wrong? Well, all the claims from the original infographic (left) are wrong! It just simply no sense if you have any basic in pharmacokinetics. Its time for the BBB scientist to deflect such woo with some science deflector shields.

First, in order to understand EOs, you have to understand their origin. To understand their origins, you have to have some basic understanding of pharmacognosy.

Pharmacognosy and EOs:

Pharmacognosy is the science that studies the chemical and biological properties of substances produced by plants and fungi. They are seasoned experts in botany, plant biology and analytical chemistry. Their main interest is to extract chemicals from different part of the plant (stem, sap, roots, leaves, flowers, fruit….), identify the substances present in such extract and identify their possible biological properties (this is often linked with ethnopharmacology, in which scientists are trying to identify the potential of some medicinal plants with their use from healers and shamans).

Plants and fungi synthesize two major classes of molecules: those involved in the primary metabolism and those involved in secondary metabolism.

Primary metabolism mostly aimed to ensure growth plant and reproduction. You can consider it as the core chemical plant. These are chemicals important for the plant function.

The second metabolism is on its own very interesting. At first, these compounds have no role in plant growth and therefore may appear useless. Indeed compounds produced from secondary metabolism are very important for the plant because these are essential for its survival. Plants evolved to have limited mobility and therefore are easy target for predators. But what plants traded out for limited mobility have indeed traded in one of the most sophisticated chemical warfare. Plants have evolutionary developed one of the most advanced and versatile chemical warfare aimed to control and deter any dangerous entities that may compete for limited resources (water, minerals, oxygen, light, CO2…).

Here are some examples of chemicals synthesized by plants secondary metabolism: Caffeine, atropine, cocaine, morphine, tetrahydrocannabinol, strychnine, nicotine, digoxin, ouabain, terpenes, cyanide, colchicine, vinblastine, paclitaxel, acetylsalicylic acid, phalloidin, forskolin, turmeric acid…….these are all products from the secondary metabolism. Many of them sounds like “poisons” and they are rightly called poisons because they can kill you at the right dose. But if you use these compounds at the right dose, these compounds can also be used to treat cancer, heart failure, glaucoma……..considering the dose makes the poison.

EOs are a particular class of chemicals, because they harbor particular chemical features. They are volatile (they belong to the superclass of volatile organic compounds or VOCs), lipophilic (soluble in fat and oils) and are odorant (this is why we can smell them). They are also capable of some biological activity.

These EOs have to be extracted from the plants via the use of organic solvent. One of the most common solvent is ethyl alcohol or ethanol (CH2OH), that is convenient organic solvent. Ethanol can help dissolve both lipophilic and hydrophilic substances. It is also has a low evaporation temperature (78ºC), allowing it to dissipate fast once on skin contact.
Another property of these compounds contained in EOs is their ability to become volatile. We can refer these compounds as volatile organic compounds (VOCs). This is why we use them as fragrance. Because they are volatile, these compounds are spread in the air and can be caught by our olfactory receptor neurons (ORNs), making what we refer a smell a smell. Smells are very powerful stimuli, even for humans. This is why we are all fond of “eau de toilette”, “eau de parfum” and all these molecular cues that can turn our reptilian brain upside-down.

EOs are very diverse by their origin and their composition. For the simplicity of this article, I will focus on the major source of EOs by their production: Citrus sirensis (sweet orange) and Mentha arvensis (mint) EOs. These are the two most prevalent sources of EOs worldwide.

Two studies that I have found listed the EO composition from these two plants.  C. sirens is  has about 50 different compounds identified, mostly classified as terpenes. M. arvensis  have about 30-40 compounds including terpenes and other organic compounds (http://www.ifrj.upm.edu.my/18%20(04)%202011/(10)IFRJ-2011-062.pdf and http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.927.8806&rep=rep1&type=pdf)

EO composition vary between cultivars, between crops, between extraction procedures. …..It means that EO by nature are anything but “pure”. EOs are therefore impure because you have a mixture of different compounds at different concentrations. It also means  that such EOs are setting the perfect storm for some drug interactions and some toxicity due to photo activation (some terpenes like limonene are know to become phototoxic following exposure to light)  or induction of an allergic reaction.

Pharmacokinetics and EOs:

In this second part, we will refute the claim brought about the penetration and tissue targeting mentioned in the infographic. The infographic have it wrong at so many different levels but two are striking: firstly, the sequence of events followed the extent of these events.

In order to understand the rebuttal, we have to understand some basic aspect of pharmacokinetics (PK). PK is the science that will tell you the fate of a chemical in your body. It will tell you how it is absorbed, how much reach the bloodstream and the tissue, how it gets detoxified and finally how it gets eliminated.
PK focuses on the fate of drugs inside our body, whereas toxicokinetics (TK) focuses on the fate of poisons and toxins inside our body.

Both follow the ADME acronym: Absorption (tegumentary/skin, intestinal/gut….), Distribution (bloodstream, tissues and brain), Metabolism (“detoxification” via chemical transformation and inactivation by the liver) and Elimination (via the liver and kidneys).

This is where the infographic has it completely wrong. It makesthe assumption that the EOs enter the brain, then the bloodstream and finally cells is just what I call “bullshit” and simply a reflection of a sheer ignorance of human anatomy and physiology. I dragged a small sketch to described the EOs ADME profile.

EssentialOilFirst, EOs have to pass the skin (or gut) barrier and diffuse all the way through the bloodstream. This operate through a passive gradient that result in the progressive loss of EOs compounds during the diffusion (depicted by the yellow arrow) into the bloodstream. This phenomenon is called “bioavailability” and investigate how much of a compound can reach the bloodstream following an administration route that is not obtained via direct injection into the bloodstream (intravenous or intra-arterial).

At the end of the day, this is the appropriate order of sequence: skin->bloodstream->brain (if you are lucky enough).

The amount of compounds contained in the EOs reaching the blood circulation remains unclear and poorly understood. But we can use some analogies with known chemicals. We will discuss the case of hydrocortisone (a topical steroid) and nicotine (a known compound capable to cross the BBB and act on the central nervous system).

Hydrocortisone is commonly used by practitioners to treat skin rashes and other irritations with a cream. The good thing about it is that such cream act topically. A thesis has documented previous studies that estimate about less than 5% of the amount of hydrocortisone applied to the skin was able to get a full ride into the kidneys (https://www.unispital-basel.ch/fileadmin/unispitalbaselch/Bereiche/Querschnittsfunktionen/Spital-Pharmazie/Diss_Pellanda.pdf). It is also telling you that a topical administration is probably not the best option for administration of a drug.

Now, there are other cases of topical administration that result in brain delivery. This is the case for nicotine and nicotine patches. These delivery systems are good in giving a good bioavailability, but yet these compounds will take some time to reach the brain. Considering the tmax (time by which a compound reaches a maximum plasma concentration), such delivery systems can only deliver nicotine with a tmax of 5-6 hours following patch application (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1364642/?page=4). You can understand that the probability of compounds contained in EOs to reach the brain within 30 seconds is impossible, unless you perforate the skull and perform an “intraventricular injection”. This is a very invasive procedure requiring a brain perforation and the insertion of a canule deep inside the brain.

Now we can argue that some drugs can reach the brain within minutes following injection. This is true for anesthetics like propofol. However propofol administration route and chemical properties are very different from EOs: they are injected via IV infusion and propofol penetration across the blood-brain barrier (BBB) is known and documented. Yet, it still takes about 4 mins for a IV infusion of propofol to achieve tmax , making the alleged claims of 22 seconds in the infographic completely bogus (https://www.fda.gov/ohrms/dockets/ac/08/briefing/2008-4354b1-01-FDA.pdf).

Even if your compounds can diffuse the skin barrier at the speed of light (100% absorption and bioavailibity) and have no metabolism (0% loss in EO compounds), you still have to demonstrate that such compounds can cross the blood-brain barrier (BBB). The BBB  blocks about 95% of chemical compounds known by humans. Therefore it is very unlikely that all these EO compounds magically fall within the 5% range.

In addition, analyzing the fate of every single chemical compound present in one EO can be an analytical nightmare even for the most seasoned analytical chemist. @SciBabe can explain you that in more details.

In conclusion, the ability of EOs to exert their biological activity beyond their skin application is simply “dead in water” and subsequently the claims posted in the infographic.

EOs and their “therapeutic claims”: the FDA warning letters
EOs may smell good but they have no scientific basis to support their claims of therapeutic use as depicted on their website. This is why the FDA has decided to enforce its authority via warning letters to two companies.

In 2014, the Food and Drug Administration (FDA) sent two warning letters to Young Living (https://www.fda.gov/iceci/enforcementactions/warningletters/2014/ucm416023.htm) and DoTerra (https://www.fda.gov/iceci/enforcementactions/warningletters/2014/ucm415809.htm) noticing them of the violation of the Food, Drug and Cosmetic Act by advertising therapeutic claims that have not been asserted. Unless you file an application in which you document with a great care the safety and the efficacy of a therapeutic agent, you have no rights to make a claim that compound X will prevent or cure cancer or other illnesses. What was true for these two companies also applies for any companies selling EOs.

Do not use EOs to treat or cure any illnesses because their therapeutic activity have not been proven by scientific methods. Worse, if misused these EOs can become dangerous poisons if swallowed  (http://www.poison.org/articles/2014-jun/essential-oils). You have been warned.

In conclusion,  EOs make great scents and fragrances to make your house smell nicely. But that should be their only application. Use them as personal fragrances with extreme precautions and avoid their swallowing and use as medicines.

[Junk Sciences] About that Dr. Neides article published in Cleveland.com

I am sure you have now heard about the infamous opinion article published by Dr. Daniel Neides, MD (Cleveland Clinic) titled “Make 2017 the year to avoid toxins (good luck) and master your domain: Words on Wellness” and published in Cleveland.com, the online site of “The Plain Dealer”, a local newspaper (http://www.cleveland.com/lyndhurst-south-euclid/index.ssf/2017/01/make_2017_the_year_to_avoid_to.html).
What was sounding first as a silly title just in time for the post-holiday “detox diets” and other scientific fallacy, quickly spinned into a conspiracy nightmare. I would not have been surprised if it came from some quack doctors that make a living on selling supplements, books and other premium services.
What was the most concerning was this article came from a medical director from a highly regarded US Health Institute, as the article was signed as “Dr. Daniel Neides is the Medical Director and Chief Operating Officer of the Cleveland Clinic Wellness Institute.
You can check for yourself but the content of this opinion article was simply frightening (I am commenting on the article as it was published on 1/8/17 at 10:52AM.
LYNDHURST, Ohio–I am tired of all the nonsense we as American citizens are being fed while big business – and the government – continue to ignore the health and well-being of the fine people in this country. Why am I all fired up, you ask?
I, like everyone else, took the advice of the Centers for Disease Control (CDC) – the government – and received a flu shot. I chose to receive the preservative free vaccine, thinking I did not want any thimerasol (i.e. mercury) that the “regular” flu vaccine contains.
Makes sense, right? Why would any of us want to be injected with mercury if it can potentially cause harm? However, what I did not realize is that the preservative-free vaccine contains formaldehyde.”
The article first sentence just set the tone: Dr. Neides has a grief about Big Gov and Big Business in general. I am mad on the government too, but not for the same reasons. We have been facing one of the most polarized election with the two parties trenched into their ideological trenches, with the average Joe in the cross-fire. We cannot have a ruling government without having a bipartisan consensus that help find a common ground to move on.
Then things gets sour as he is fingerpointing on the CDC, again using the allusion of Big Gov for his grief on this institution. It quickly turns into a anti-vaccine diatribe, with spelling errors (it is “thimerosal/thiomersal” not “thimerasol”). Not only Dr.Neides fail to name his grief correctly (and you cannot blame autocorrect on that one) but also makes a scientifically fallacy by associating “thimerasol” as mercury. As previously mentioned in my blog, there is a huge difference between mercury as an element (Hg) and the organomercury compounds such as methylmercury (CH3Hg) and ethylmercury (CH3CH2Hg). Considering organic chemistry part of the pre-requisite courses for any medical school, this type of mistake is not acceptable.
In a common anti-vaccine move, Dr. Neides quickly played the “goalpost moving” tactic. If you first argument failed following its refutation by facts, move to another target and play the same spiel. If it is not thimerosal, then it is the formaldehyde. If it is not the formaldehyde, then it is the aluminum……..
We are not yet at the end of the first paragraph and already Dr. Neides has taken a dangerous slippery slope in pseudoscientific claims. He will continue into the “appeal to authority” fallacy, claiming the government is condoning the global population health. This is why the government has agencies like the Department of Health and Human Services (DHHS), the Food and Drug Administration (FDA), the Center of Diseases Control (CDC) and the Environmental Protection Agency (EPA). They have doing their job, could be better but it could be worse. Considering our next government line-up, we will likely soon regret how “red tapes” is sometimes needed.
But lets go to the second part of his article with this very interesting quote:
We live in a toxic soup. There are over 80,000 chemicals used in various industries country-wide. There are over 2,000 new chemicals being introduced annually. We breathe in these chemicals through exhaust, eat them in our processed foods ( just look at the labels that have 20 or 30 ingredients and good luck pronouncing their names), textiles (clothing, bedding, furniture), and personal care products, including make-up, deodorant, shampoos, and soaps.“.
Of course we are living in a chemical soup, but not all of them are toxic. There are some that are neutral, some are toxic (this is what we refer as poisons and toxins and are extensively studied by a discipline called toxicology) and some are even beneficial (these are identified by a a discipline called pharmacology).
Authorities are constantly testing these chemicals for 40 years now and there is a huge effort from international agencies to provide a formidable toxicological database to document the toxicity of any existing chemical inventoried.
A side-note, the quote “good luck pronouncing their names” sounds eerily familiar to a quote from a certain Babe “If a third grader can’t pronounce it, don’t eat it“. I am guessing I should ditch the addition of “alpha-D-glucopyrannosyl-1,4-alpha-D-fructofurannoside” from my morning cup of Joe. Thats by the way the biochemical name for sugar.
It is followed by a mishmash of different terms that just sound like a nightmare for anyone teaching pharmacokinetics to healthcare professional.
Toxins accumulate in our fat cells if they are not eliminated and interrupt normal bodily functions. Your body should be a finely tuned machine with all of the organ systems working in concert together. But when toxins disrupt normal function, problems can occur. Those problems include cancers, auto-immune diseases, neurologic problems like autism, ADHD, and Parkinson’s disease, and the most prevalent chronic diseases like obesity, diabetes, and heart disease (note: so high blood pressure is not a chronic disease, then?).
Why are we so sick in 2017 despite the best access to healthcare? The body has wonderful built-in systems to help us detoxify. The liver and kidneys try to do an exceptional job keeping up with filtering out the “stuff” (toxins included) we don’t need. Our skin – the largest organ in the body – will release toxins in the form of perspiration (what did they teach you in med school? How did you graduated from your biochemistry class?). Our breath will release toxins with each exhalation (same “boingboing” moment). When our gut is healthy and our microbiome (100 trillion organisms that live in our intestinal tract, within our airway, and on our skin) intact, our bowel movements help rid unwanted toxins.
I like to think of our detoxification system as a big bucket. As long as the toxic soup stays within the bucket, our body can naturally eliminate what we don’t need and help us live at the highest quality of life. But what happens when the bucket starts to overflow – which is exactly what many of us have been facing our entire lives? The body may not have the capacity to eliminate our current exposures and THAT IS WHEN BAD THINGS START TO OCCUR.”
We, as animals, have evolved to deal with many chemicals in nature that can have a possible threat to use. In pharmacokinetics, we refer to ADME for Absorption, Distribution, Metabolism and Elimination.
Absorption of compounds is limited for water-soluble compounds, they need a special door (called solute carriers) to enter inside your body. Now some (and many drugs in general) are fat-soluble (we call them lipophilic compounds) and can diffuse passively through the intestinal barrier and get into the bloodstream. Even if these drugs can make it through the gut, they have to face a firs-pass metabolism by the liver (we will discuss later about it) and deactivate some if not all of them. Those who were not affected by the first-pass are ongoing a distribution phase. Distribution occurs by the spread of the compounds throughout the central compartment (blood circulation). From this compartment, compounds can diffuse into peripheral (tissue) compartments. One of this compartment is named the “deep storage compartment” (a nice word for your fat tissue) that can retain some lipophilic drugs. As long as you ingest them, you can accumulate these compounds. Once you stop, these compounds move from the periphery back into the central compartment. We know which compounds can store (the lipophilic ones) and such feature is considered in toxicological studies to define if a compound is toxic or not. After circulating, compounds are perfusing through the liver and undergo of series of chemical reactions called metabolism. Metabolism is ensured by a series of enzymes grouped into two phases: Phase I enzymes (called cytochrome P450s or CYPs) and Phase II (conjugating) enzymes. Phase I enzymes transform compounds from a fat-soluble into a water-soluble compounds. Phase II enzymes tag these compounds to ease their elimination. Finally, the elimination phase is the one by which the body get rid of these compounds. There are two major routes: hepatobiliary (liver via the bile duct and eliminated via feces) and renal (kidneys).
Again, the fallacious claim that skin and respiration are considered as major elimination routes is simply outrageously wrong, especially considering from a healthcare professional.
The third part is directly playing the classical anti vaccine tropes “vaccines cause autism” that have debunked again and again.  I will not into it, there are great resources about what this claim is purely wrong and not supported by science.
But I just wanted to add my take on Hepatitis B (HepB) and newborns vaccination. The general public has a global misconception about HepB being only an STD. Indeed it is not. HepB is the same type of virus than HIV when we talk about mode of transmission: a blood-borne pathogen (in addition to its sexual transmission). This is a vaccine we highly recommend to any staff working in lab research and handling any blood or blood-based products. Because there is always a risk of HepB transmission via accidental cut and exposure to blood, this is even supported by being an “healthy carrier” (you have circulating HepB in your blood but you are not having any liver condition). it is better safe than sorry, considering than HepB can lead to severe liver pathology such as cirrhosis.
Finally the fourth part was fairly alarming. You may have heard about Orac, a seasoned MD and oncologist that is doing a great job in his website “www.sciencebasedmedicine.org“. Orac has been raising his concern that pseudoscience and quackery has been percolating inside mainstream medicine, using names like “functional medicine”, “integrative medicine” or “holistic medicine” as a wolf with a sheep cloth, appearing harmless.
He cites then a quote from a colleague named Jessica Hutchins, MD and IFM-certified (certification from a certain Institute of Functional Medicine) as described as the following:
In a 2015 article in U.S. News and World Report (Link to her article), Jessica Hutchins, M.D., IFM certified practitioner, states , “Information on eating toxin-free food and pushing food manufacturers to stop using harmful ingredients can be found at foodbabe.com. When we vote with our dollars by choosing to buy products that are sustainably produced and chemical-free, we actively shape the market place. Help change the way [loved ones] nourish their precious bodies, starting with yourself as an example.
Using Vani Hari (aka the FoodBabe) as a source of information and as a reference for health advices, from an MD, is simply insane! “What did they teach you at the IFM?”. At first glance, the certification program provided by the IFM seems legitimate (https://www.functionalmedicine.org/certification_program/About/) however one single item appear intriguing:
Comprehensive assessment of the fundamental physiological systems that organize the key clinical imbalances and are affected by the mental-emotional-spiritual core: Assimilation, Communication, Defense and repair, Transport, Energy, Structural Integrity, Biotransformation and elimination
Until now, we have little or no evidence of prayers and thoughts work in healing someone, lesser than any evidence faith healer healed someone solely on their spiritual powers. Also the presence of chiropractors as instructors (and we know how well chiropractors have a scientific training) is also disturbing.

In conclusion, 2017 comes in with a bang as I feared: a year in which evidence, facts are discredited by gut-feeling, smooth talks and snake oil sellers.

Post-scriptum: when you have woo peddlers lauding your article, you know that our article is a complete #NaturalNonSense (courtesy: Chow Babe).
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[Junk Sciences/BBB] Does bone broth leads to a leaky brain? (aka the stupid, it burns!) 

Everyone should have recovered from the post-Thanksgiving food comatose (no, it was not due to the turkey’s tryptophan), the post-election discussion and the Blck Friday melee fight for that 55″ UHDTV that is justin time to play on your XB1S with a 4K resolution.
Seem the woo is never running short of idea, piling up fallacy over another fallacy as iHop piled up pancake in a post-Thanksgiving breakfast ride.
The latest fallacious claim cames from a “nutrition blog” claiming that bone broth, leads to elevated glutamate levels following bone digestion (!), such glutamate results into a leaky gut. Leaky gut means utomtically leaky brain, letting glutamate flows into the brain as boiling pasta water flows through the hole of the pasta strainer and ultimatelly leads to neurological disorders, with a special mention of neurodevelopmental disorders such as ADHD and ASD, because these are two conditions that scares parents as hell nd therefore makes a juicy alibi to milk on parents withtheir precious $$$ and torture kids with dubious or completely scam treatment (including bleach enema, putting kids on casein-free/gluten-free diet, hyperbaric oxygen chamber or chelation treatment).
Because I am the one claiming the claim of this blog post is bogus, it is my duty (and my pleasure as a BBB Scientist) to debunk it.

1. About the bone broth….

In the post, the author first claim that bone broth prepation (slow cooking) results in a dissolution of the tendrils and connective tisues to dissolve, as cited as the following
So what’s the dark side of Bone Broth? It lies in the “long” part of the “slow and low and long” cook time. Bone Broth simmers at a low temperature for many hours, long enough to allow the connective tissues to dissolve and the minerals to be drawn into the broth.Thats partially true. Bone broth is composed by two major type of tissue: connective (bones, tendrils, muscles) and non-connective tissue (basically the bone marrow). The preparation of bone broth will primarily dissolve the bone marrow (rich in bone marrow stem cells) that gives that yummy flavor in dishes, my fond being the French “Pot-Au-Feu” (the yummiest soup during the holiday season). The cooking process will tenderize the meat but very unlikely to dissolve bones and tendrils.
To dissolve bones and tendrils, you need something……very acidic. Only an extreme acidic and hot environment will dissolve bones and tendrils.
Therefore her first claim is not fully backed by science. Now let’s assume you have some connective tissue that solubilize, you will have mostly collagen dissolved. Collagen is the main protein involved in most connective tissues. The author claims that such processing releases tons of glutamic acid (that is also glutamate) as described here “The result of such long cook times is a tremendous amount of glutamic acid. And that’s the dark side of Bone Broth
.Collagen is a polymer made of a same repeat motif Glycine-X-Y, with X and Y being often proline and hydroxyproline (the hydroxylation of proline into hydroxyproline is made by prolylhydroxylase, using vitamin C as a co-enzyme. This is why scurvy (lack of vitamin C) is characterized by defect in connective tissues).
It comes with different flavors depending on the function (collagen type I  being the predominant form in connective tissue; collagen II found in tendrils; Collagen III in cartilages and bones; Collagen IV in basement membranes……). This second fact discredits her claim that collagen will increase the amount of glutamate.

2. What is the blood-brain barrier (BBB) and how glutamate is processed?
After giving her culinary explanation, the author jumps into explaining glutamate into her own terms: “As you may or may not know, glutamic acid and glutamate are highly regulated by the human brain“. You can already note the red herring about the author as she treats glutamic acid and glutamate as two distinct chemical entities.  This is an obvious lack of understading of biochemistry  as glutamate is the the ionized (salt) form of glutamic acid. This is a typical misconception that I see with folks peddling on woo (folate/folic acid anyone?). It also raises concern about the credentials of someone making dubious health claims. Garbage in, garbage out.
Not only the author makes blatant mistakes but also makes fallacious associations as mentioned here If you have a leaky brain, as those with a leaky gut often do, high amounts of glutamic acid can trigger seizures if you’re prone to them. Yes, seizures. It can also trigger other neurological symptoms you may already be sensitive to, including brain fog, migraine headaches, dramatic mood swings, stimming, and nervous tics, to name a few.
I have been discussing over and over about the BBB in my blog, so I will not repeat over and over again. I will keep it simple. The BBB is what we call a component of the neurovascular unit (a combination of the cardiovascular and nervous system) that is designed to protect the brain from any interferences from the periphery. Therefore the environment in which the brain bathes in is strictly controlled, with a defined composition. The BBB provides two kind of barrier: a physical and a chemical barrier supported by the brain microvascular endothelial cells (BMECs) lining the inner side of blood vessels. They provide a tight blood-brain interface that strictly regulates what comes in and out the brain. The physical barrier is harbored by the presence of tight junction complexes, whereas the chemical barrier acts as a selective filter that pick and choose which molecules goes into the brain through an array of transporters, represented by the super-family of solute carriers (SLCs). Each solute carriers are adapted to let the entrance of specific molecules (for instance SLC2A1 lets glucose enters the brain, SLC16A1 lets T3 thyroid hormone enters the brain….).
In many aspects, BMECs share a lot of features than intestinal cells. But a leaky gut and a leaky BBB cannot explain her claims.  In order to understand how her claim is bogus, I will argument using a recent review by Hawkins RA and Vina JR (the first author has a publication records in regards of glutamate transport accross the BBB). If glutamate
However, glutamate is firstly poorly transported inside the brain because it remains trapped inside the BBB . Thats already hit her claim.
If we follow her logic, we are facing a massive increase in glutamate level in the food uptake (due to the broth). Because you have a leaky gut (a concept I have been already explained in my previous blog as still debated and mostly inaccurate in terms of diseases), you have your body flooded with glutamate and ultimatelly flooding your brain with glutamate. This makes sense only if you have a deficit. Chemicals flows into two compartment following the Ficks law, it diffuse from the most concentrated to the least concentrated compartment. Because it is considered as a non-essential amino acid (your body knows how to build it up from other molecules), the brain is not in need for glutamate and can produce its own. Glutamate plasma concentrations range from 50-100microM, brain concentrations ranges from 10’000-12’000 microM. We are talking about 200 times more in the brain than in the plasma. Therefore, you need to achieve a concentration higher in the brain to expect to see any changes. Thats simply impossible and destroy the rest of her argument!

3. But, but…….I have a leaky brain! (No you have not).
The idea of a leaky brain is simply the most ridiculous claim. If your brain was indeed leaky, you would not be here reading this post. You will be six feet under, dead from a massive brain edema.
Leaky brain happens when you have a disruption of the BBB, in particulat from the tight junctions lining BMECs. This results into an onset of “vasogenic cerebral edema” and in an unrestricted entrance of water and ions inside the brain.
This happens if you are experiencing high-altitude mountain sickness (in particular high-altitude cerebral edema), stroke or brain trauma (traumatic brain injury). Because the brain is contained inside a rigid structure (skull), it cannot expand and swell as you would experience if you experience and injury in the periphery. This results in an increase of an intracranial pressure (ICP) and brain injury by mechanical stress (imaging squishing a sponge in your hand) that leads to neuronal cell death. Untreated, brain edema is fatal. However, there are cases of cerebral microbleeds that do not induce cerebral edema formation. This could be something that can be considered as “leaky brain” but their intensity and extend is a far cry of the former word meaning. 
Thus, the concept of leaky brain is completely fallacious as it is taken out of context and grossly exagerrated. 

[Pharmacology/Junk Sciences] About the FDA recall on Dr. Hyland’s homeopathic tooth relief….

You may have heard about the recent FDA recall for Dr. Hyland’s homeopathic teeth relief and any products similar to it as listed in their report below (http://www.fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm523468.htm).

There are 10 children deaths and 400 adverse events reported with the association of this homeopathic product. Homeopathy has always lauded itself as “natural, effective and harmless”. Even when it was faced with the lack of efficacy, proponents of homeopathy always use the “what’s the harm?” as a defense. Seems that not only it will not treat your ailment. Worse it can kill you. Why? Simple pharmacology.

A tenet of pharmacology is known since Paracelsus with his famous citation “Every substance is poison. No substance is no poison. The dose and only the dose makes a substance a poison”. Even the natural products are poison. You see, Mother Nature in all its grandeur has developed a certain affinity for developing some of the most toxic substances designed to kill living species. Because of evolutionary constraints, plants have been very good as being “chemical plants” and use chemical warfare as a method of invasion and defense.

Now, you can use this poison effect to treat certain conditions. If you find the right dose, you may treat a condition. This is how pharmacology works. Under a certain dose, there is no effect. We only start to see some biological activity after a certain dose. This is what we call the Minimum Efficacy Concentration (MEC). On the other hand, if you reach concentrations high enough, then you start to hit on off-targets and develop the minimum toxic concentration (MTC). This is where we start to see side effects and if the doses are high enough you will see adverse and toxic effects.

Homeopathy is by definition an unproven therapy because its principles defy common laws of physics and chemistry. Because it is relying on extreme dilutions to explain its activity, the amount of active substance is so low that it can be compared to dilute one single molecule into a swimming pool.

In this case, we don’t exactly know what compounds and what amount of each alkaloids are extracted. We know the total amount is 0.0000000000003% or 0.3 pico-grams per 100mL of solution or 100g tablets.

belladonna

That amount is simply ridiculously low. First, we have to assume that its PO administration will result in a 100% bioavailability (that is probably false and surely below). Second, you should be able to detect the compound in plasma samples. At that level (pg), the odds of detection are very low if not non-different from background. Therefore the odds of having this single molecule finds its target inside a body is almost zero (and the probability is something like 0.0000000000000000000000000000001%).

Yet, homeopathic treatment are all starting from raw extract (usually an hydro-alcoholic solution as extracting solvent) called “tintura matter”. Thats a concentrate and it contains indeed active substances in a high concentration. The nature and quantity of active substance in it are not known and not even reported since homeopathic products are not falling under the FDA supervision (you can thanks the DHSEA provision for that). Basically, no one (even the manufacturer) will tell exactly what and how much of active compounds are inside this tintura matter.

This raises a problem because Mother Nature has been excelling in producing poisons, what only matter is the dose. In the case of the teething relief product, the main culprit appears Atropa belladonna, a common poisonous plant that get easily confused with edible berries.  One of the classical substances purified from Atropa is atropine, an acetylcholine muscarinic receptor blocker. Because Atropa belladonna is known for its poisonous activity, we had some disaster in waiting that was cooking here. Anyone working with drugs will tell you how dangerous it is to overdose on an active substance.

Acetylcholine is an important neurotransmitter in mammalian cells, with peripheral (heart, muscles, lungs….) and central actions. Acetylcholine is a key target of organophosphates and sarin gas, as these agents block its degradation by acetylcholine esterases (this is why you have atropine administered as a side treatment for poisoning).

However, atropine can and will have severe side effects if ingested in a poisoning event (see this case report of accidental ingestion of Belladona: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3361210/)

So whats exactly happened in this situation? We dont know and only now the FDA can provide clues. The FDA is toothless and cannot execute any order unless there are fatality cases involved. On their page, they are aware of the anticholinergic side effects of it but minimized the risk (that logically make sense since the amount is ridiculously low)

belladonna2

At this point, I speculate that something wrong likely happened with these homeopathic products. My speculation is something went wrong with the preparations, resulting in inconsistant extraction and effective concentrations in tintura matter. Because homeopathy are not ruled by FDA, they do not have to follow the stringent quality control and good manufacturing practices (GMP) imposed on pharmaceutical companies. So we have here a production that was on a free ride without any oversight on the QC. Now you can easily imagine that we may have ended up on with overdosed preparations.

Considering the special population (pediatrics), this risk of overdosing was even more amplified. One thing we consider when we develop a drug and prescribe a drug is the benefits/risk ratio. If the risk or adverse effects are too high, the drug cannot be used because it overcomes the benefits.

In that case, not only the benefits were quasi-null, we provided access to a product that was deemed “safe because natural” but indeed has a risk of adverse effect. Now we have likely some precedent in directly demonstrating the danger of poisoning with homeopathic products. This is already the case with essential oils (there are cases of children poisonings with Eos) and now we have the same with homeopathic products.