Friday, December 27, 2013

Genetically Silenced Apple. Science or beliefs?

 A biotechnology melodrama in Tweets.

Hendrick Goltzins The Fall of Man 


Have you heard of the non-browning  Arctic Apple engineered to market sliced apples and reduce waste? The GMO apple is galaxies away from fruit Adam and Eve tasted in the Garden of Eden.

 It's worth reading some science going into its creation to learn if we feel at ease taking a bite and tasting this modern fruit....dwelling a bit on the tree of knowledge good or evil, science or belief?

If, like me,  you have ever felt like some GMO scientists sound more evangelical than scientific by the time you are done reading this modern  Twitter drama there will be less doubt your feelings are legitimate.

Though it might give you a little headache if you aren't  a scientist, I think it'll be worth your time. Knowledge is like that. Biotechnology science is as fascinating as it gets, even if a little complicated.

 It could be a fun exercise in separating science from illusions of science-beliefs.

If you aren't science minded skip the details. Ask yourself a basic question "am I seeing statements fitting better in the science or religion category"? The fog might lift a just a little.

Science 101: 
Evidence-based
Customarily presented in a form of shiny citations to data ( non-scientists might find annoying).
Usually containing raw numbers, science symbols, charts or graphs.
Core tenet:  questioning assumptions.:
 " What is the evidence for my theory/   hypothesis /  belief     ???????"


Religion--belief based.
No data, proof, or evidence required.*
Science citations inevitably absent.
Assertions made requiring zero proof.
* Don't recall the last time they showed numbers or graphs at a religious ceremony.
 It is a good thing, too -can't remember the last spiritual, transcendental graph I've seen, do you? 


  



According to the New York Times, these apples are neither wanted by consumers nor apple growers. 


I have some questions about testing before taking a bite, don't you? [1] [7]
Were precise profiling techniques used e.g. transcriptomics, proteomics, metabolomics ( the "omics")?
Any feeding trials conducted on laboratory rats?
The answers appear to be "None" & "No".


If approved, the Apple will be one of the very few products designed with silencing technology intended for use as food for people, and it will have evaded proper safety testing.  [9]








Unlike the common herbicide-tolerant and insecticidal GMOs,  the GMO apple was not created by the addition of genes. (+) 


A genetic engineering technique called RNAi or RNA interference was used.  Double-stranded RNA (dsRNA) which binds messenger RNA (mRNA) preventing the production of the browning enzyme; PPO  was introduced.  (*)


So as promised here comes the Tweeter-drama. As the USDA was wrapping up its public comment period on the Arctic apple advocates  took to Twitter, where the following ensued and was  storify'ed  for you.


 Darryl Hannah tweeted a link to her followers encouraging them  to voice their disapproval

 Predictably "scientists"  flocked to "educate the masses".


The website is a  glossy promotional site without a single science citation to be found.


So I pressed the scientist & Co again, and again, and again, and again to post links to science on off-target effects (OTE).
 You see it is very well known that today's silencing technology leads to unpredictable unintended effects causing surprises.
  
 " When designing therapeutic short-interfering RNAs (siRNAs), off-target effects (OTEs) are usually predicted by computational quantification of messenger RNAs (mRNAs) that contain matches to the siRNA seed sequence in their 3' UTRs ( untranslated regions) [..]

 We observed no correlation between the number of computationally predicted OTEs and the actual number of seed-dependent OTEs (P=0.76). On average only 20.5% of actual transcriptional OTEs were seed-dependent (i.e., predicted)." [6]


Sometimes as discovered in a study on bees - scientists silenced genes they didn't  even know existed!  [3]

 A study on insecticidal RNAi reported that dsRNA  engineered to kill insects produced unintended secondary dsRNA with different sequences and different targets than originally planned which did the insects in. [4] A similar process of generation of secondary dsRNA has been described in plants.[8]

Off-target effects causing unpredictable gene silencing are far from conspiratorial. They are discussed by USDA's scientists [5]




 Suzy, the scientist claims  there have been 10yrs of testing which, while true,  is misleading because  they were years of agronomic research and development, rather than safety testing.

  Not a single study on any experimental animal-lab rat or human- exposed to this silenced fruit  is anywhere to find. Not a single "omics" study ( transcriptomics, metabolomics or proteomics) [7]  is cited in spite of multiple requests.

 Thus saying  they are nutritionally equivalent is a leap of faith- not rigorous science.







The tweet below is where it got really interesting!







1. She is unaware that a study recently completed at Rutgers University demonstrating short dsRNA (microRNA)  in the viscera of rabbits fed a therapeutic (antiviral) microRNA engineered tomato.  The scientists filed a patent for plant-based therapeutic applications using this technology.  Thus, her declarative statement that microRNA won't survive digestion is steeped in a brew of ignorance  and is scientifically untrue.





2. It should be clear to readers that absence of PPO ( browning enzyme) and its gene in humans is irrelevant in the context of off-target effects in the plant itself or animals ingesting the silencing gene carrying apples.

3. She appears unaware of a study demonstrating microRNA from bacteria, fungi and even insects has been detected in human plasma. How did they get there? Did she not know or has she not bothered to ask?  [2]. Oh, yeah--Monsanto insists microRNA can't be absorbed, and is still arguing that microRNA are a contaminant to avoid safety testing its crops.

4. But, there is not a single study anywhere in the literature showing rates of degradation of double-stranded RNA at Ph 2!   Never mind in the range of gastric pH's (2-5)  in adult humans.

 So Suzy, the Scientist, can Not back up any of her claims. Her only refuge would be to call me names, which she already tried, but I skipped that part because ad hominem statements starting with " you are...X, Y, ...Z" are so common in these GMO  "conversations/ religious conversions."

 Thus, I pressed the scientists to show me a study on degradation of dsRNA at ph2. Here is what happened.





Dr Suzy the Scientist posts a  link to a study on chemical hydrolysis of ribonucleic acids. Not only is it completely irrelevant to dsRNA, but the hydrolysis reaction takes place after an hour of boiling. How many people do you know who stew their stomach contents at 220F for an hour?

As a brilliant scientist  watching this little drama unfold commented: "Funny how they so adamantly claim these things but cannot show any data."


The stuff sounds pretty indestructible now.  Of course, it isn't, because we have lots of enzymes called pancreatic RNases to digest RNA, Suzy the Scientist evidently forgot about.



OK!  Finally! After days of unscientific nonsense, we arrive where we should have started --at our first science citation. Or rather  a citation to a study on pancreatic ribonucleases (RNases), enzymes that degrade RNA, which, by the way,  have little to do with chemical degradation at pH 2.







First, no data on how fast the dsRNA disappears from the stomach, nor anywhere else. It was bacterial dsRNA, not plant dsRNA.  Nor was this a paper measuring acid stability, but enzymatic stability.

No relevance to how fast the dsRNA disappears and whether or not it is rendered inert during digestion time under physiological conditions.
(Thanks to Dr Heinemann!)


I did eventually get a citation to a second study on degradation of dsRNA in the stomach kindly posted by Dr David Tribe aka GMOpundit.


The trouble is that Dr Tribe's study actually showed  that  dsRNA is stable for extended periods of time relative to time in stomach (10 to >24 hours) and is  not more unstable at pHs that might be relevant to human digestion (5)

Fig 3  at >ph 6.8. dsRNA detectable up to 10 hours at all pHs tested.
Detectable at 24 hours at all except pH 6.8 (and this appears to be an anomaly).
Fig 4 they test to only as low as pH 5, not pH 2  Still present after 24 hours at the LOWEST tested pH of 5!

Curiously enough, Dr Tribe stated "  Simply put, the relevant wheat RNA molecules will be rapidly digested in the gut to harmless, simple nutrients." on his blog discussing silenced wheat.

It turns out there might not have been any science evidence to back up that assumption/ hypothesis/belief.

As far as I can see in this little melodrama, NO studies showing these novel stable RNA molecules that we've never eaten- showing it gets degraded in the human gastro-intestinal tract - were cited at all.










I am of the mindset now that some "scientists" either have never questioned their deeply held assumptions, maybe since high school. Someone way smarter than myself once said:  "assumptions are called ASSumptions because they make ASSes out of us". Or they are willfully selling illusions they know nothing in common with science, and much more in common with religious beliefs.

Unfortunately, the unconscionable denial of risks of RNAi is hindering promising medical research!    http://www.boulderweekly.com/article-12640-muzzled-by-monsanto.html, which is most definitely very tragic.

References:

1. Securing the safety of genetic modification
Dr Jack A. Heinemann
 http://theconversation.com/securing-the-safety-of-genetic-modification-13102


2. The complex exogenous RNA spectra in human plasma: an interface with human gut biota? 
 PLos One 2012;7(12):e51009. doi: 10.1371/journal.pone.0051009. Epub 2012 Dec 10.
 http://www.ncbi.nlm.nih.gov/pubmed/23251414


3.  Non-Target Effects of Green Fluorescent Protein (GFP)-Derived Double-Stranded RNA (dsRNA-GFP) Used in Honey Bee RNA Interference (RNAi) Assays
Francis M. F. Nunes 1,†,* email, Aline C. Aleixo 1,†,* email, Angel R. Barchuk 2email, Ana D. Bomtorin 1, Christina M. Grozinger 3email and Zilá L. P. Simões
 http://www.mdpi.com/2075-4450/4/1/90


4. Nat Biotechnol 2007 Nov;25(11):1322-6. Epub 2007 Nov 4.
Control of coleopteran insect pests through RNA interference.
Baum JA, Bogaert T, Clinton W, Heck GR, Feldmann P, Ilagan O, Johnson S, Plaetinck G, Munyikwa T, Pleau M, Vaughn T, Roberts J.
 http://www.ncbi.nlm.nih.gov/pubmed/17982443

5.  RNAi-Based Insecticidal Crops:  Potential Effects on Nontarget Species
Jonathan G. LundGren and Jian J. duan
 http://www.aibs.org/bioscience-press-releases/resources/Lundgren.pdf


6. Br.J.Cancer 2013 Feb 5;108(2):450-60. doi: 10.1038/bjc.2012.564. Epub 2013 Jan 8.Lack of correlation between predicted and actual off-target effects of short-interfering RNAstargeting the human papillomavirus type 16 E7 oncogene.
Hanning JE, Saini HK, Murray MJ, van Dongen S, Davis MP, Barker EM, Ward DM, Scarpini CG, Enright AJ, Pett MR, Coleman N.
 http://www.ncbi.nlm.nih.gov/pubmed/23299538

7.  Environment International 37 (2011) 1285–1293
Review Molecular profiling — a tool for addressing emerging gaps in the comparative risk
assessment of GMOs.
Jack A. Heinemann a,b,⁎, Brigitta Kurenbach a,b, David Quist b
a School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand b GenØk – Centre for Biosafety, Tromsø, Norway
Table 1
Expert views on profiling.
Method Positive perspectives on profiling
High throughput sequencing. High-throughput sequencing proved to be a powerful and quantitative method to sample transcriptomes deeply at maximal resolution. In contrast to hybridization, sequencing showed little, if any, background noise and was sensitive enough to detect
widespread transcription in N90% of the genome, including traces of RNAs that were not robustly transcribed or [were] rapidly degraded. (Wilhelm et al., 2008)
Proteomics Therefore, DIGE [DIfference Gel Electrophoresis] is highly appropriate for comparative profiling of knockout, transgenic, or isogenic germplasm as well as defined pharmacological or stress induced responses as recent reports suggest. Another important consideration is that quantitative studies using 2-D gels in general, and 2-D DIGE in particular, can be performed with any plant
species and are not restricted to plants with sequenced genomes. (Thelen and Peck, 2007)
Proteomics Inter-experimental reproducibility is not good using 2-dimensional polyacrylamide gel electrophoresis approaches, however, an excellent differential display technique (DIGE) allows multiple samples to be compared on the same gel and is a powerful tool in biomarker discovery for laboratory exposure studies which is quantifiable. (Van Aggelen et al., 2010)
Metabolomics [Catchpole et al. (2005)] demonstrated the compositional similarity between GM and non-GM potatoes using mass spectrometric fingerprinting as a primary screen, which was then amended by detailed quantitative profiling analyses. This approach (i.e. combining non-targeted and targeted analytical methods) appears feasible even for screening large numbers of transgenic plants, and the application of both supervised and unsupervised data analysis techniques ensures independency from statistical bias. (Rischer and Oksman-Caldentey, 2006)
Metabolomics [The Catchpole et al. (2005) study was] the first report of a large scale metabolomic analysis of field grown GM potato plants, and it highlights the importance of analytical technology and appropriate data analysis for the safety assessment of GM crops. (Colquhoun et al., 2006)
Metabolomics The work reviewed here demonstrates that the tools that are currently available for metabolite profiling are mature and robust enough to facilitate their use in the investigation of biological processes.(Schauer and Fernie, 2006)
Metabolomics In particular, an impressive number of natural foods, spices and beverages have already been the subject of detailed metabolomic-based component analysis, including milk, grapes, tomatoes and tomato juice, rhubarb, beer, celery seeds, coriander as well as many other herbs and spices. These analyses used a combination of NMR, GC–MS, LC–MS and CE techniques
to identify up to 100 different phytochemicals or 200 different carbohydrates in selected fruit, vegetable or beverage samples. (Wishart, 2008)
Mass spectrometry [It is now] possible to choose from a variety of techniques, such as flow injection electrospray ionisation mass spectrometry (FIEMS), Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR), which facilitate rapid fingerprinting of crude extracts, [to provide profiles of chemical compositions of GM plants].(Rischer andOksman-Caldentey, 2006)
http://www.ncbi.nlm.nih.gov/pubmed/21624662

 8. http://www.hfsp.org/frontier-science/awardees-articles/triggering-secondary-sirna-production-plants-role-mirna-structure

9. A comparative evaluation of the regulation of GM crops or products containing dsRNA and suggested improvements to risk assessments
  • a School of Biological Sciences, University of Canterbury, Christchurch, New Zealand
  • b Centre for Integrated Research in Biosafety, University of Canterbury, Christchurch, New Zealand
  • c Crop Science Department, Federal University of Santa Catarina, Florianópolis, Brazil
  • d Health and the Environment, School of the Environment, Flinders University, Australia
  • e Institute of Health and Environmental Research, Adelaide, South Australia, Australia
  Open Access
http://www.sciencedirect.com/science/article/pii/S0160412013000494
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Footnotes: 

(+) Technically, the Arctic Apple was created by inserting a new piece of DNA. The new piece is designed to be transcribed like any other gene making a single stranded (ss)RNA. The ssRNA has internal complementarity, so once it is made it forms a stem loop structure (see Fig 1C of the paper I link to above). This structure is then a substrate for DICER (or Drosha) and can subsequently enter the RISC complex that directs it to mRNA targets (intended or not). [9]  See cool video here

In theory, it is not necessary to introduce the DNA to get the effect. Animals take up the dsRNA directly and there are plans to make dsRNA-based pesticides that are directly taken up by plants or animals. Ironically, those pesticides may not need regulatory approval because some are arguing that unless there is a DNA step, as in the Apple, it is no longer genetic engineering! Another attempt to subvert safety testing using semantics.

(*)   mRNA which guides production of protein should not be confused with miRNA ( microRNA), which are an example of a short double stranded (ds) silencing RNA.










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Thursday, October 3, 2013

The Land of Confusion

   Who Let The Mites OUT? Allergies.... Again

 


How some GMOs might make them worse

 

 



Man....I wish I didn't suck at immunology.  The last time I studied the subject, the Gipper was President.
A one-hit-wonder  was topping the charts having us walk like an Egyptian. The juiciest affair of the era wasn't between a President and an aide wearing a blue dress,  or a Twitter flurry of penis images- it was the Iran-Contra affair involving an exchange of weaponry.  Genesis described the era best-  it really was a land of confusion.




 Needless to say, those long forgotten lessons did not make me an immunologist, and I  hate playing one on the net.   But a  recent Twitter conversation with Nathanael Johnson, a food reporter writing a long series on GMOs on Grist,  prompted me to dig around  for whatever remnants of immunology I've not yet forgotten- to explain the scientific reasons I am seeing allergic reactions to GMO ingredients in my patients, clinically- in real life. 

So, lets review some basics. 

In people atopy causes hay fever, asthma and eczema, while food allergies are an undeclared emerging epidemic.

The job of the immune system is to detect and destroy "bugs" out to kill us while leaving our own tissues alone, known as tolerance.  Allergies are seen when the immune system gets "confused" and overreacts to harmless stuff like pollen or dust mites (allergen/ antigen)  reacting to them as if they were lethal invaders. When the immune system declares nuclear war on the body, the result is a life-threatening anaphylactic reaction. 

An allergic reaction typically occurs when an antibody known as IgE binds the antigen, activating mast cells to release inflammatory chemicals, the most famous of which is histamine.
Common anti-histamines like Benadryl work by preventing their release.  Thus, when testing for allergies is done, what immunologists are usually testing is the antibody IgE.

 Did you know that an allergy to birch tree pollen can cause allergies to apples and other fruits?  It is true. This is due to an antigen is known as bet v 1. And allergy to cockroaches or dust mites could make you more prone to allergies to shrimp due to a shared tropomyosin.   And if you were to develop an allergy to latex you might become allergic to kiwis and chestnuts.

  • Cross-reactivity explains the reasons for the overlap of symptoms between inhalant/contact allergies known as atopy and food allergies.  

 Cross- reactivity refers to the ability of the IgE antibody to react with more than one allergen/antigen from different sources due to shared antigenic components, such as those between birch pollen and fruit, or grasses and grains, or dust mites and shrimp. Good list of cross-reacting allergens.

Cross reactions arise because the binding site on the allergen-- known as an epitope-- the business end of the allergen made up of a chain of folded amino acids--fits the IgE antibody,  like a lock and key in three dimensions.


Phew...... not too bad, right?

Now that we have some basic immunology figured out,  what does any of this have to do with GMOs?
 

We will get to it in just a minute.
First, I'd like to introduce you to some microscopic critters billions of whom share our homes, our beds, couches and live on your pet's kibble. They are known as dust mites. Nasty looking critters, aren't they? Another type of mite called storage mites,  just as ugly as these, also live on cereals and your pets' dry food.

Dermatophagoides pteronyssinus Wikipedia

Because house dust mites are microscopic the general feeling about them among clients could be "How much harm can they cause my pet or me?"
 In fact, the most common allergen in dogs and cats (and in people, too) is dust mites. Anyone who is allergic to dust mites can attest to the miserable symptoms this condition can cause. It seems our domestication of dogs and cats (i.e., bringing them into our households and beds) also succeeded in sensitizing them to this common allergen.....so; they suffer from them just like we do.
  • So, what if.. these mites are not only living on the food but are actually in the food? 


And this is where genetically modified foods (GMOs) come in. No, I am not saying that eating a genetically modified organism (GMO) means you are eating microscopic mite body parts.


  • But...  there is a distinct possibility that a segment of a dust mite allergen might have been spliced into Round Up Ready GMO foods. 



And this is why.

The most common commercialized GMOs are Round Up resistant: corn, soy, canola, sugar beets and alfalfa. They were engineered to be herbicide resistant by insertion of a  transgene known as Cp4 EPSPS which produces a protein EPSPS  (Enoyl Pyruvyl Shikimate Phosphate Synthase), isolated from a bacteria found growing in an herbicide waste tank,  which rendered it resistant to the herbicide- Round Up -Agrobacterium CP4.

Insertion of exogenous, new and foreign DNA into plants raises concerns about unintended effects such as the creation of novel allergens.  The World Health Organization (WHO) gathered together global experts in allergies and published guidelines on screening GMOs for known allergens in 2001.

The  very first step in the decision  tree WHO experts established screening GMOs for novel allergens is this fundamental rule.  " if a  new gene product  spliced in shares 6 amino acids ( protein building blocks) with known allergens, such a  match  triggers further allergy testing  on  serum from  25  people with an allergy to that substance."

It turns out that Cp4 EPSPS shares a window of similarity of  7 amino acids with common house dust mites, an allergen known as der. p.7.  ( abbreviation of  Dermatophagoides pteronyssinus).  
It isn't the only one.
If you look at Table 4 and Table 5 in the link below, you will notice other potential allergenic matches between new genes being spliced in and known allergens, which I haven't checked.   The decision tree is also graphically illustrated in the article linked below.


 Screening of transgenic proteins expressed in transgenic food crops for the presence of short amino acid sequences identical to potential, IgE – binding linear epitopes of allergens






















 This brings me back to the conversation with Nathanael and  a link  he provided to studies on genetically modified foods conducted thus far which  help answer the following question:

"did the companies commercializing these crops conduct proper studies to make sure that the transgene was not allergenic based on the guidelines established by the WHO"?  Did they even perform the most basic preliminary steps to screen out allergens? 

 And the answer is: no, not really.





 While GMO advocates quote these authoritative bodies to support their safety, as you will soon find out--the basic safety guidelines established by these bodies are not followed! 

Ruling out cross-reactivity between the dust mite allergen and the new protein coded by the transgene, an ELISA inhibition assay is required on 25 serum samples from patients with high titers to dust mites-- according to a global panel of allergy experts assembled by the World Health Organization.  Since allergies to dust mites are so incredibly common, it should be exceedingly easy to locate serum samples.

 Unfortunately, a targeted screen utilizing   inhibition ELISA was performed on a pooled sample of 4  in one of the studies cited by Nathanael. Interestingly, it did show minimal cross-reactivity.  But pooling a sample reduces its accuracy while  a sample of four has a very low probability of detecting cross-reactivity.

Additionally, the studies cited were performed on patients in Korea, Portugal and Japan---countries where GMOs are neither as prevalent as in the US nor sold unlabeled. There is not a single study done on a statistically significant number of samples (>10) from Americans with the longest and highest level of exposure to GMOs. 

Thus,  if you, your child or your pet is sensitized to mites, dust mites or storage mites,   the allergies might be exacerbated by foods containing some or all Round Up Ready crops:  soy, corn, canola, sugar beets ( beet pulp). Remember, with allergies it isn't the quantity or the amount--it is exposure to an allergen.

  • Unfortunately, it takes only a tiny amount of mite allergen to elicit an allergic reaction in our pets and people. 
  • Allergic cross-reactivity is often encountered as a reaction without prior exposure.  A very common public misunderstanding is that people and pets express allergies on the first time they are exposed to an allergen. This is very rare, as normally sensitization over a long period is required to have allergy symptoms. 
If you or your pet are sensitized to dust mites of which we have an unbelievable amount in Long Beach California,  or storage mites, does it mean that they or you will automatically be allergic to these foods?   No, it does not.  Even if there is cross- reactivity between the new gene in these foods and mites, cross-reactivity alone does not mean a clinical allergy. It isn't as simple as that. But being aware will allow you to watch yourself or your allergic pet for exacerbation of allergy symptoms with exposure to these ingredients or improvement when they are eliminated, and acting accordingly.


Oh, and please--  if any real immunologists are reading this I'd   appreciate your help! I hope you've enjoyed this little journey through the land of confusion. 


------------------------------------------------------------------------------------------------------------

Footnotes: 

 [1] Email I sent to Nathanael- I never received a response. 


Hello, Nathanael.
I believe that GMOs are allergenic based on seeing patients for many years who improve when they are removed from their diets or deteriorate when added to their diets.Allergies are veterinarians' bread & butter out here in dusty Southern California and after 24years of practicing I think I've gotten pretty good at them.
 As I shared with my friend Kevin Folta, I see a case or two a week, whose allergic flare ups, I suspect, are triggered by GMOs. The last one was 2wks ago--the owner switched food, and the dog broke out on his face and feet within 48hrs. The only difference was that the new food had beet pulp, most likely RR. Someone who isn't aware of GMOs would never notice the connection and most vets are clueless about them.  Plus they are much easier to treat with immunosuppressive drugs, then dig around for the cause.  I suspect, but don't know, that the same level of ignorance is also true of the human medical profession--which is even worse because specialization so compartmentalizes it.
Of course, my observations are anecdotal, but if I am right, there is a whole lot of suffering and pain these foods could be contributing to. The immune system is incredibly complicated, there is more we don't know than we do, and there isn't a fine line between an allergy, seen as a nuisance, and devastating inflammatory/ autoimmune diseases in the genetically-vulnerable.
The studies in your link are a mess. They are not standardized; they are poorly done and hard to follow, and none of them do a serum test on 25 patients with allergies to Dermatophagoides pteronyssinus (der. p.). Some do skin prick tests rather than serum tests; others are made up of small sample sizes (10).  There is no standardization of methodology.
 The safety tests of GMOs are a mess, in general. It is like that example you gave off different people feeling different parts of an elephant. So while you are reading a study that examines the elephant's trunk or his tusk, it says absolutely nothing about the whole darn beast.  
The best that can be said about them is that they are pilot studies and that studies on a much higher number of people need to be done.
The Goodman affair I linked on Genetic literacy was to show clearly that Dr. Goodman, who is the director of the biotech allergy database, as well as the new editor of the journal most of the GMO literature is published in, I believe, could not provide a study which fulfills the WHO-established allergy testing standards. Neither could Rod Herman of Dow, who rewrote the GMO- allergy rules, likewise, in the comments in your series.  Either because they do not exist. Or they do exist, since it is a relatively simple test, and it showed that EPSPS and dust mites do cross react, and these companies know it.

I don't know how journalism works, but you do have two biotech people saying on the internet print "record" that the WHO guidelines for allergy testing are not being followed. Ask yourself if that is good enough considering that CP4 EPSPS is in most of our human and animal food and just about everyone and their aunts, uncles & cousin are allergic to dust mites.


I am not sure whether Grist is really allowing you to do genuine investigative journalism, because I noticed ads for Dows cotton in your series. Don't know, but expect, that media outlets have to pay their expenses with advertisers dollars and the biotech companies are in the midst of a media blitzkrieg.


Having said all that, I hope you are a "good guy", might have some room to maneuver and can do some public good.

If you want to know why I am spending this much time on the subject it is really very simple. I have unique training. As a veterinarian I took an oath which obligates me to prevent pain and suffering and watch out for public health. I don't take oaths lightly, and I feel an allegiance to the taxpayers of the state of California who spent a quarter million to educate me. Besides, if  someone, somewhere, doesn't do something,  Monsanto et. al. will destroy the field of biotechnology--they have already turned it into a dirty word.   
Thanks for listening.
Please do your best to do some good. With all the smoke & mirrors, combined with how very complicated this subject really is, some intelligent dogged reporting is sorely needed.
Thanks very much,
Ena
  

Sadly, for the American public,  there are very few trustworthy  journalists on the GMO beat abiding by the   Journalism Code of Ethics.

 Society of Professional Journalists explainer on  journalism Ethics:
Members of the Society of Professional Journalists believe that public enlightenment is the forerunner of justice and the foundation of democracy. The duty of the journalist is to further those ends by seeking truth and providing a fair and comprehensive account of events and issues. Conscientious journalists from all media and specialties strive to serve the public with thoroughness and honesty. Professional integrity is the cornerstone of a journalist's credibility.


[2] Le Monde article on Dr. Goodman in charge of the allergen database. 







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