Sunday, September 23, 2012

Lost Rebuttal from Dr. Ena

Dr. Ena Valikov is a Veterinarian from Huntington Beach, CA.  She frequently comments on posts, usually those regarding transgenic technologies, and presents coherent arguments that elevate the discussion.  She has a background in biochemistry so she speaks science well and can discuss the literature.

Yesterday morning my gmail account posted several responses to my September 21 post. There were two there from Dr. Ena.  I was excited to read them and prepare my responses. Yet when I looked at the comment section of the article one of her comments was not there.  Instead, there was an appropriately cynical comment from Dr. Ena about censoring the comments.

I have no idea what happened or where her comment disappeared to.  However, I was disappointed and upset for several reasons.  First, I appreciate an informed rebuttal because I am the first to admit, I might be wrong.  I'm glad to consider all evidence in my synthesis.  Second, I would never, and have never censored a comment. On one of my YouTube posts someone made rude and offensive comments about one of my student's foreign accents.  I left it, and pointed out its ignorance. This is a marketplace of ideas and to be a censoring or dismissive gatekeeper is the stuff of activism, not science.

I'm posting here Dr. Ena's lost comment.  I can't seem to find her as I don't have her actual email address, so I hope this is acceptable (I'll take this down if you don't want it posted, Ena).  I just feel awful, I don't like how it taints the perception of communication in this forum.

So here I post Dr. Ena's points in response to my Sept 21 post, and her arguments supporting Seralini's recent work.  My comments will appear below in the Comments section.

Ena Valikov has left a new comment on your post "Rats, Tumors and Critical Assessment of Science": 

(KF) Ena, a replicated study would be great, unfortunately Seralini's stuff never is replicated.
Meaning that you don't have a single LONG TERM STUDY examining laboratory animals for Long Term Chronic effects.

No, I don't think this study is trash, because I know mammary tumors to be estrogen sensitive. The study demonstrated elevated estradiol levels in both males and females and even proposed a mechanism of action by which EPSPS can alter estradiol levels. 

As to the control groups and experimental group : why is there a study on biofortified composed of 15 rats, which none of you objected to?
Prima facie evidence that a phytocystatin for transgenic plant resistance to nematodes is not a toxic risk in the human diet.http://www.ncbi.nlm.nih.gov/pubmed/14747684

It is a study on 15 rats fed purified extract rather then the genetically modified rice… and the only biochemical/ hematological patient data actually published is

TABLE 1
Summary of results from the toxicological study of male Sprague-Dawley rats administered the cystatin OcIΔD861

OcIΔD86 [mg/(kg · d)]
0 0.1 1 10
Food intake, g
    d 3–7 25.04 ± 0.94 25.80 ± 0.90 26.82 ± 1.48a 25.36 ± 1.42
    d 7–10 26.06 ± 0.40 25.68 ± 1.32 27.74 ± 1.43a 25.82 ± 0.98
Organ weight
    Cecum (empty), g 0.305 ± 0.033 0.334 ± 0.057 0.32 ± 0.013 0.35 ± 0.041a
    Liver, g 3.12 ± 0.16 3.00 ± 0.072a 3.02 ± 0.090a 3.04 ± 0.12
Serum analysis
    Potassium, mmol/L 4.51 ± 0.27 4.78 ± 0.318 4.66 ± 0.251 4.95 ± 0.82a
    Urea, mmol/L 6.58 ± 0.82 5.61 ± 0.67b 6.09 ± 0.616 6.24 ± 0.887
    Creatinine, μmol/L 39.0 ± 2.83 39.7 ± 3.02 41.0 ± 1.66 41.5 ± 3.21a
    γ-Glutamyl transferase, U/L 0.07 ± 0.067 0.42 ± 0.67a 0.13 ± 0.11 0.18 ± 0.13

——————————————————————————————
Here is what a blood panel should actually look like:
Test Result Reference Range
ALK. PHOSPHATASE 294 10 – 150 U/L HIGH
ALT (SGPT) 57 5 – 107 U/L
AST (SGOT) 25 5 – 55 U/L
CK 171 10 – 200 U/L
GGT 4 0 – 14 U/L
AMYLASE 344 450 – 1240 U/L LOW
LIPASE 397 100 – 750 U/L
ALBUMIN 3.9 2.5 – 4.0 g/dL
TOTAL PROTEIN 8.4 5.1 – 7.8 g/dL HIGH
GLOBULIN 4.5 2.1 – 4.5 g/dL
TOTAL BILIRUBIN 0.2 0.0 – 0.4 mg/dL
DIRECT BILIRUBIN 0.1 0.0 – 0.2 mg/dL
BUN 34 7 – 27 mg/dL HIGH
CREATININE 1.2 0.4 – 1.8 mg/dL
CHOLESTEROL 336 112 – 328 mg/dL HIGH
GLUCOSE 131 60 – 125 mg/dL HIGH
CALCIUM 11.0 8.2 – 12.4 mg/dL
PHOSPHORUS 8.3 2.1 – 6.3 mg/dL HIGH
TCO2 (BICARBONATE) 25 17 – 24 mEq/L HIGH
CHLORIDE 87 105 – 115 mEq/L LOW
POTASSIUM 4.3 4.0 – 5.6 mEq/L
SODIUM 144 141 – 156 mEq/L
A/G RATIO 0.9 0.6 – 1.6
B/C RATIO 28.3
INDIRECT BILIRUBIN 0.1 0 – 0.3 mg/dL
TRIGLYCERIDE 98 20 – 150 mg/dL
NA/K RATIO 33 27 – 40
HEMOLYSIS INDEX (1) N
LIPEMIA INDEX (2) N
ANION GAP 36 12 – 24 mEq/L HIGH

WBC 26.7 5.7 – 16.3 K/uL HIGH
RBC 8.03 5.5 – 8.5 M/uL
HGB 19.1 12 – 18 g/dL HIGH
HCT 52.0 37 – 55 %
MCV 65 60 – 77 fL
MCH 23.8 19.5 – 26.0 pg
MCHC 36.7 32 – 36 g/dL HIGH
NEUTROPHIL SEG 80 60 – 77 % HIGH
NEUTROPHIL BANDS 5 0 – 3 % HIGH
LYMPHOCYTES 4 12 – 30 % LOW
MONOCYTES 11 3 – 10 % HIGH
EOSINOPHIL 0 2 – 10 % LOW
BASOPHIL 0 0 – 1 %
AUTO PLATELET 725 164 – 510 K/uL HIGH
PLATELET COMMENTS
PLATELETS APPEAR INCREASED.

ABSOLUTE NEUTROPHIL SEG 21360 3000 – 11500 /uL
ABSOLUTE NEUTROPHIL BAND 1335 0 – 300 /uL
ABSOLUTE LYMPHOCYTE 1068 1000 – 4800 /uL
ABSOLUTE MONOCYTE 2937 150 – 1350 /uL
ABSOLUTE EOSINOPHIL 0 100 – 1250 /uL
ABSOLUTE BASOPHIL 0 0 – 100 /uL

SENIOR PROFILE W/ TRIG : T4
Test Result Reference Range
T4 (1) 2.5 1.0 – 4.0 ug/dL


The study’s limitations are quite obvious: its duration is 21 days and N=15, not to mention that the rats weren’t fed the genetically engineered rice, but rather the isolated protein (which is not equivalent to the whole food).

Can you please explain how findings in 15 rats fed this GMO for 21 days imply safety in millions of people, eating the stuff for decades?

Because from my vantage point, the only time you really care about control group size, or statistics--is when a study comes out suggestive of long term harm. You know ... the kind you would never catch, if you only study the GMO for the required 90 days. 



Posted by Ena Valikov to Illumination at September 21, 2012 10:47 PM


28 comments:

Kevin M. Folta said...

Ena, Let's get right to your question on the bottom of the page:

Can you please explain how findings in 15 rats fed this GMO for 21 days imply safety in millions of people, eating the stuff for decades?

The study was not designed to justify feeding transgenic food to people. The study was from Howard Atkinson, the world authority in cystains (protease inhibitors that can affect nematode -and other pest- digestion).

Atkinson and colleagues have performed many tests on cystatin mechanism of action, efficacy in the lab as an anti-nematode agent, and even environmental impacts of the transgene.

The goal of this study was to test if there was any sign of harm in animals from consumption of the agent. It was not a test to conclusively show that this was great to feed babies.

It also was not performed in a commercialized transgenic food. In fact, cystatins are still not used in transgenic plants commercially.

The first words of the title say Prima facia, admitting up front that this is a first look, a pilot test in a new area of research. No evidence of harm was observed in these conditions. This is important because if there was a problem it is best caught early in a small study.

On the other hand, Seralini et al present an argument to overturn an industry. As revealed by their conference call and the verbiage of their text, they claim a conclusive, final, solid link between GMOs and tumors in SD rats. This extraordinary claim requires extraordinary evidence, large numbers, proper statistics, and no hidden controls.

As a scientific reviewer we have to ask first, what is the hypothesis being tested? Are the experiments appropriate to test they hypothesis? Do the data and statistical treatments reject or support the hypothesis?

Atkinson's paper does this. Seralini's does not.

That is the difference between the two papers, even though both show no compelling evidence of harm.

Kevin M. Folta said...

I just found this, thought it was excellent.

"Forbes notes one independent academic source pleased to see the Seralini work. The Statistical Laboratory at the University of Cambridge was "grateful for the authors for publishing this paper, as it provides a fine case study for teaching a statistics class about poor design, analysis and reporting." Adding they "shall start using it immediately."

Ena Valikov said...

Epigenetic Events Associated with Breast Cancer and Their Prevention
by Dietary Components Targeting the Epigenome
Shabana I. Khan,†,‡ Pranapda Aumsuwan,†,§ Ikhlas A. Khan,†,‡,|| Larry A. Walker,†,§,^ and
Asok K. Dasmahapatra

caffeic acid partially inhibited the methylation of RARβ
promoter/MCF-7, MDA-MB-23197
RARβ/MCF-7,
MDA-MB-23197


http://pubs.acs.org/doi/pdf/10.1021/tx200378c

dogctor@yahoo.com

:)

Ena Valikov said...

Roles of microRNAs in breast cancer
Over the past few years, miRNA profiling studies have led to the identification of miRNAs that are aberrantly expressed in human breast cancer. The function of only a handful of these miRNAs in breast cancer has been investigated. As in other cancers, some miRNAs can function as tumor suppressors and other miRNAs as oncogenes. Thus, tumor formation may arise from a reduction or deletion of a tumor suppressor miRNA and/or amplification or overexpression of an oncogenic miRNA. In addition, tumor metastasis may be promoted by enhanced expression of prometastatic and/or downregulation of antimetastatic miRNAs. The functions of these miRNAs in breast tumor progression and metastasis are discussed below.

http://breast-cancer-research.com/content/12/2/201

PS: the fist very long post disappeared in the internet vapor, again!

Ena Valikov said...

So, Kevin-- your blog has eaten a pretty long response TWICE.

It showed up for a few minutes, twice and then disappeared--twice.

Tired of playing.

And the robo-captions are impossible to read, to boot.

Ena Valikov said...

You are still NOT censoring, COMRADE?

Pathetic, really!

Ena Valikov said...

Rebuttal 2.0. Kevin.


http://www.science20.com/science_20/blog/gm_maize_causes_tumors_rats_here_how_experts_responded-94259#comment-123041

Kevin Folta said...

Hi Ena, first, thanks for being a good sport and participating again. I honestly don't know what happened.

Unfortunately I'm completely slammed at the moment. I'd love to respond, but it won't be until Saturday! I will address your points. Thanks.

Michael said...

"You are still NOT censoring, COMRADE?

Pathetic, really!"

Kevin, why you continue to engage this person is beyond me, you pathetic communist.

Ena Valikov said...

Ena Valikov ena1101961@gmail.com
5:39 PM (17 hours ago)

to Kevin
It's cool Kevin. I know what it's like being busy- don't sweat it.

I look forward to your response when you have time.

Cheers,

Ena



http://www.youtube.com/watch?v=JmcA9LIIXWw

and oldie but goodie ;-)

Kevin M. Folta said...

Ena, I'm absolutely not censoring. I don't know what is happening. If you endure the hassle of copy/paste of your responses and email them to me I'll be glad to post them for discussion.

Ena Valikov said...

Part 1/2
Can you please explain how findings in 15 rats fed this GMO for 21 days imply safety in millions of people, eating the stuff for decades?

The answer to the question is “ it can't".


The points are simple:
There is not a single valid blinded long term clinical controlled feeding trial in the literature.


The second point is that if you are going to criticize Seralini, you might want to apply the same standards of logical and scientific consistency. Plucking the Only long term trial on record in the history of OECD biotechnology regulation ( which in spite of all its flaws the study satisfied) and criticizing the control group, while staying mum about the similarly grossly unbalanced statistics of the GMO study cited below. There are bloggers who are critical of it because it isn't blinded, while there is not a Single blinded GMO safety study Anywhere in the literature.

Extraordinary claims deserve extraordinary evidence except where SAFETY ASSURANCE STUDIES by Hammond et al. are concerned. Assuring us that the crap is safe for millions to eat for generations is an extraordinary claim backed by No evidence at all. When 90 day rodent tests ( based on OECD regulations no one complained about in the past), some of which are performed on 400 rats of which there are 80 experimental animals, of whom partial data is published for 10-20, ya might as well just call the evidence “ oh hell, just trust me”.

Results of a 90-day safety assurance study with rats fed grain
from corn rootworm-protected corn
B. Hammond a,*, J. Lemen a, R. Dudek a, D. Ward a, C. Jiang a, M. Nemeth a, J. Burns b
a Monsanto Company, 800 North Lindbergh Blvd., St Louis, MO 63167, United States
b Covance Laboratories, Inc., 9200 Leesburg Pike, Vienna, VA 22182-1699, United States
Received 1 June 2005; accepted 22 June 2005

And I don't see anything addressing this fact on biofortifiedd, where it turns out there isn't a single long term controlled feeding trial satisfying even basic medical standards is to be found, just like the rest of literature. Please cite One.


Did Serallini show long term harm? you betcha! The rats fed GE corn or GE corn with Round Up, or Round Up died before the controls (Figure 1)

Is it statistically significant?

Unlikely without a single p-value reported in the entire study. However, previous studies already documented kidney and liver pathology, making the shorter life-span not even remotely surprising. The breast cancer finding is a complicated one because there are dozens of pathways and feedbacks that drive breast cancer – estrogen, estrogen receptors and antagonists, aromatase inhibitors and so on. In a world without politicized science one would start by replicating the estradiol levels reported in rodents fed the transgenic corn since estradiol is mitogenic and examining whether the overexpression of ESPS could be driving synthesis of other aromatic estrogenic metabolites.

Ena Valikov said...

Part 2/2

Finally microRNA's. Karl at biofortified began the censorship game months ago. I admit that my email to him wasn't fuzzy and nice, but the censorship started when I wished to discuss the implications of the rice miRNA study. The ideas propagated at biofortified are so fragile, hatched by scientists so thin- skinned, that he might as well be guarding the delicate things in a medieval castle behind an alligator-filled-moat.

Do I realize we have been eating microRNAs for millennium? You betcha.

Does that mean I should be comfortable with rushing engineering plants (some of whom might be deploying this new knock-out technology) without thorough controlled feeding trials and without publishing sequences of novel mi RNA created by transgenic technology? The rice mi RNA study changes old paradigms. Acknowledging that a study which documented that plant miRNAs aren't digested, are absorbed into the blood stream and are regulating genes across kingdoms, including human organs, should lead to re-thinking the old rules such as EPSPS is a bacterial-plant enzyme catalyzing a metabolic pathway wich doesn't exist in humans, therefore it can't possibly do anything to a mammal... not true anymore.
When classical insect miRNA are found to play a role in Human breast cancer.
let-7f, a tumor-suppressor mi RNA which targeted the aromatase gene (CYP19A1) expression.
http://www.ncbi.nlm.nih.gov/pubmed/22407818. the old paradigms such as Bt is a toxin specific to insect guts can not affect mammalian guts lacking those specific receptors....likewise needs to change.
You might engineer neutraceutical, medicinal miRNAs or oncogenic miRNA. The old " don't look, don't tell" wont work. There are too many miRNA's playing a role in breast cancer as well as all sorts of diseases to feel comfortable stating with certainty that there is no mechanism ( endocrine or epigenetic)by which the trangenic corn can not raise rates of breast cancer.

And since you dedicated a blog to me, and I love music--here is one you might not have heard before ( no gambling in this one) some say to forward to 1.08min
http://www.youtube.com/watch?v=0YTh1Wsqo2c


Ena

Kevin Folta said...

Ena, I just don't get your veracity in defense of this work. Now that this has blossomed to allegations of scientific misconduct and fraud, many scientists, even those not engaged in the GMO discussion are now tearing this thing to pieces. I'll address your points, but I'll tell you up front, you spend too much time with misdirection. My original post was about the Seralini paper and a scientific post-hoc criticism of points missed by other analyses. But I'll work laterally to touch your points.

There are no reports that show GMO safety, because science cannot prove that something, anything, is safe. We can only show evidence of harm using a given model and set of tests, over a given time. That’s it. You keep bringing up safety. Do they ever test any new variety, organic culture system, novel hybrid for evidence of harm? Never.

>>> The second point is that if you are going to criticize Seralini…

I’ll have to look at the Hammond paper. If there’s something wrong with it I’ll put it up next time. However, my analysis of Seralini’s work stands on its own. It’s clear agenda and intent are deplorable. That’s my point. That stands regardless of what others publish before/after.

>>>>Extraordinary claims deserve extraordinary evidence….

Again, nobody says it is safe, just evidence of harm. I’m itchy to see Hammond. I’m not at work now, so I can’t access. However, I’d be surprised if they say it is “safe” and if they do they are wrong. Again, this shows your bias. Breeding programs NEVER test new varieties, despite moving thousands of genes. No safety assessment, because it is impossible to test everything. There is no plausible mechanism for harm, and no evidence of harm.

>>>And I don't see anything addressing this fact on biofortified….

Here’s the problem. You want some long-term study to show something is safe. What is “safe”? What does that mean? If someone spent millions of dollars to hire postdocs and do the experiment to test that GM is dangerous, what are the endpoints? What do you measure? Who would fund this with no preliminary data showing harm and no plausible mechanism that can suggest harm? No funding agency would fund such trash. That’s not science, that’s paranoia. Plus, if they did it and tested a dozen metrics, you’d still say they didn’t do enough… because you can’t prove something is safe. There always will be another reason to complain about studies that weren’t done.

>>>>Did Serallini show long term harm? you betcha! The rats fed GE corn or GE corn with Round Up, or Round Up died before the controls (Figure 1)

Ena, the fact that you are compelled by the Seralini paper shows that you have made up your mind, and know nothing about experimental design. You betcha? If I flip a coin and it comes up heads three times in a row, can we conclude that the coin has a “head” on both sides? Maybe according to you and Seralini. The data are meaningless if the experimental design is inadequate. Period.

>>>Is it statistically significant?

NO. It Is not. And you can’t say that previous reports from the same lab reinforce this. If someone else, anyone other than the lab in an anti-GMO institute where the lead author sells books and movies and his results are never replicated outside of his own lab… not compelling.
And when controls are not show, eg. Figure 3, it really exposes intent.


>>>> estradiol is mitogenic and examining the overexpression of ESPS could be driving synthesis of other aromatic estrogenic metabolites.

But they can’t just hand-wave and then it is so. Maybe you buy it, but scientists like me have an evidence threshold. You have to show it. Do the experiment. This is why this paper is in a low-caliber journal.

Kevin Folta said...

Ok, your second comments.

>>>Finally microRNA's. Karl at biofortified began the censorship,,,,

I know Karl pretty well. He’s a good guy, super knowledgeable and a solid scientist. I doubt there was censorship, meaning that he was suppressing ideas. I don’t know. I hope you are not implying that I am not giving you a fair forum.

>>>>Do I realize we have been eating microRNAs for millennium? You betcha.

More than a millennium... try since the dawn of eukaryotes probably. First, the rice miRNA study is a year old and no follow ups have come out. That bothers me. I thought it was a really cool paper and neat idea, but had some questions. I hoped that this would open a new area of science and cross-kingdom interaction/signaling. That would have been great.

However, as of right now, it does NOT “change paradigms”. Even if it is 100% true, then why would transgenic food be the target? Plants make natural antisense RNAs, so do fungi. Plants might make more of them when grown hydroponically, or when sprayed with rotenone in an organic plot. Plants might make more of them if the products are hot, refrigerated, not fertilized enough, fertilized too much…. THE POSSIBILITIES ARE ENDLESS! So instead of getting upset about transgenics, get upset about food. As soon as you are calling for the testing of everything then we can talk about singling out a technology.

And the food you eat is not sterile. You consume bacterial and fungal RNA every day by the bucket and none of that has ever been tested. Moreover, the miRNA content of the blood has been well characterized—literally hundreds of papers. In all of these surveys why are dietary/bacteria/fungal miRNAs not found? I dunno. Maybe only that one survives digestion and is detectable in the serum. In other words, it is not a complex soup of plant miRNA.

>>>>When classical insect miRNA are found to play a role in Human breast cancer.

Let7f an insect miRNA? Not according to the reference you cited. It showed that aromatase inhibitors used in breast cancer therapies induce Let7f in vitro and it decreases aromatase levels. Even if there is a same-sequence miRNA in insects, how does it get into humans at levels that are pharmacologically relevant?

>>>>. the old paradigms such as Bt is a toxin specific to insect guts can not affect mammalian guts lacking those specific receptors....likewise needs to change.

Why? You have evidence to the contrary?

>>>> You might engineer neutraceutical, medicinal miRNAs or oncogenic miRNA. The old " don't look, don't tell" wont work. There are too many miRNA's playing a role in breast cancer as well as all sorts of diseases.....

Ena, sure, that’s a testable hypothesis. But why do you just target trangenics? Bt is one sequence. Are you against Bt being used directly on organic crops? Probably not. You can jam the Bt sequence through a number of programs and it will tell you if it even makes miRNAs. I’ll do that. But if you feel that dietary RNA can somehow be processed, transported and sequestered in physiologically relevant levels, then the burden of proof is on you, not me. It is just kooky. I can dream up longshot ideas, I do every day. But even this would be unfundable by any agency without significant preliminary findings.

The Dire Straits song you posted was an instrumental and there were no words at 1:08.

Ena Valikov said...

Quickie Kevin, cause it is the beginning of the month and I've got bills and payroll to deal with, on top of the doctoring. Will be back later in the week to follow up.

(Ena, I just don't get your veracity in defense of this work.)
I am a truth seeker operating on this basic premise, like all medical professionals are:
Primum non nocere
From Wikipedia, the free encyclopedia
Primum non nocere is a Latin phrase that means "first, do no harm".

(Now that this has blossomed to allegations of scientific misconduct and fraud, many scientists, even those not engaged in the GMO discussion are now tearing this thing to pieces)

If they want to see fraud they should read all the Hammond papers.

400 rats, results reported for 10-20, crucial data is absent, pathological conclusions are : nephropathy and hepatobiliary disease in under 90 days.

Isolating the only long term feeding trial without comparing it to the literature titled : SAFETY ASSURANCE Study, would be twisted warped spin, rather than genuine science.

{science cannot prove that something, anything, is safe} science can certainly compare effects on metabolic organs, estradiol levels, upregulation and down regulation of proteins and genes in metabolic organs of subjects ingesting the stuff.....and show it to be as safe, less safe or equally as safe.

Do you have such as controlled blinded long term feeding trial on rats, cats, dogs, or people?

What long term scientific evidence do you actually have demonstrating the transgenic crop's effects in the experimental subjects ingesting it?


>>>> estradiol is mitogenic and examining the overexpression of ESPS could be driving synthesis of other aromatic estrogenic metabolites.<
But they can’t just hand-wave and then it is so. Maybe you buy it, but scientists like me have an evidence threshold.

This is where your myopia shows. Cats, dogs and people do not synthesize aromatic branched chain amino acids. If there is an alternative pathway through which this transgene interacts with your endocrine system--and you discover the mechanism, and engage your " glass is half full" part of the brain, you might see an entry point of intervention to reduce rates of breast cancer or other chronic maladies. At that point I'll help you sell the food.

Plants contain miraculous compounds we could use more of.... why not focus on that?


{But why do you just target trangenics?} I do not target transgenic. I target dishonest scientists.
I've got human recombinant insulin and interferon in my frige at the hospital. They are clearly labeled as products of transgenic technology. Same with erythropoietin we use on cats with kidney disease, same with Filgastrim used on patients with myelodepression due to chemotherapy.

I am not a square peg to be pounded into a round hole.


{ You can jam the Bt sequence through a number of programs and it will tell you if it even makes miRNAs.} I would rather you jam the entire EPSPS transgene with all its bacterial, viral, petunia components into that computer, along with all the flanking sequences adjoining however- many copies were inserted, and compare them to the miRNAs associated with branched chain aromatic amino acids and miRNA involved with estrogen receptors, aromatase inhibitors, or other mechanisms promoting of breast cancer. I lost a friend to breast cancer-- one of the most inspiration people I know. You, Kevin, could be on the side of history reducing breast cancer rates, or on the side potentially increasing it. Which do you see yourself on?

See you.

Another great song to inspire you to be on the side of science that helps women not suffer from breast cancer.
http://www.youtube.com/watch?v=BdDD9JX-Tyg


Ena Valikov said...

Seralini:
 
As the total  chemical composition of the GM maize cannot be measured in details, the use of substantial equivalence is insufficient to highlight
potential unknown toxins and therefore cannot replace long-term
animal feeding trials for GMOs.
A cause of the effects of the effects could be that the NK603 GM maize used in this study is engineered to overexpress a modified version of the Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) (Hammond et al., 2004) allowing the R tolerance. The modified EPSPS
is not inhibited by glyphosate by contrast to the wild enzyme. This
enzyme is known to drive the first step of aromatic amino acid biosynthesis in the plant shikimate pathway; in addition estrogenic
isoflavones and their glycosides are also products of this pathway
(Duke et al., 2003). They were not disturbed in our study. By contrast, the levels of caffeic and ferulic acids in the GM diets, which are also secondary metabolites from this pathway, but not always measured in regulatory tests, are significantly reduced. This may lower their protective effects against carcinogenesis and even
mammalian tumors (Kuenzig et al., 1984; Baskaran et al., 2010).
Moreover, these phenolic acids and in particular ferulic acid may
modulate estrogen receptors or the estrogenic pathway in mammalian cells (Chang et al., 2006). This does not exclude the action
of other unknown metabolites. This explanation also corresponds
to the fact that the observed effects of NK603 and R are not additive and reached a threshold. This implies that both the NK603
maize and R may cause hormonal disturbances in the same
biochemical and physiological pathway
 
 
One more thing. I  appreciate the offer you made to send me articles behind pay walls. I accessed the following two,
 
ferulic and caffeic acids, page 128-129
 
http://samples.jbpub.com/9780763764012/64012_Ch08_0148_rev.pdf
 
 
 
Antiproliferative and apoptotic effects of selective phenolic acids on T47D human breast cancer cells: potential mechanisms of action
Marilena Kampa,1 Vassilia-Ismini Alexaki,1 George Notas,2 Artemissia-Phoebe Nifli,1 Anastassia Nistikaki,1Anastassia Hatzoglou,1 Efstathia Bakogeorgou,1 Elena Kouimtzoglou,3 George Blekas,4 Dimitrios Boskou,4 Achille Gravanis,3 and Elias Castanas1
Author information ► Article notes ► Copyright and License information ►
This article has been cited by other articles in PMC.
Go to:
Abstract
Introduction
The oncoprotective role of food-derived polyphenol antioxidants has been described but the implicated mechanisms are not yet clear. In addition to polyphenols, phenolic acids, found at high concentrations in a number of plants, possess antioxidant action. The main phenolic acids found in foods are derivatives of 4-hydroxybenzoic acid and 4-hydroxycinnamic acid.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC400651/

but would appreciate if you could send me the other articles I can’t access.
 

Thanks!
 
 

Ena Valikov said...

Articles I would like to read.

Chemopreventive potential of ferulic acid in 7,12-dimethylbenz[a]anthracene-induced mammary carcinogenesis in Sprague–Dawley rats
·         Nagarethinam Baskaran, 
·         Shanmugam Manoharan, , 
·         Subramanian Balakrishnan, 
·         Pachaiappan Pugalendhi
·         Department of Biochemistry & Biotechnology, Faculty of Science, Annamalai University, Annamalainagar – 608 002, Tamil Nadu, India
·         http://dx.doi.org/10.1016/j.ejphar.2010.03.054, How to Cite or Link Using DOI
·         Permissions & Reprints
 
 
2.  Lipophilic caffeic and ferulic acid derivatives presenting cytotoxicity against human breast cancercells.
Serafim TL, Carvalho FS, Marques MP, Calheiros R, Silva T, Garrido J, Milhazes N, Borges F, Roleira F, Silva ET, Holy J, Oliveira PJ.
Source

In the present work, lipophilic caffeic and ferulic acid derivatives were synthesized, and their cytotoxicity on cultured breast cancer cells was compared. A total of six compounds were initially evaluated: caffeic acid (CA), hexyl caffeate (HC), caffeoylhexylamide (HCA), ferulic acid (FA), hexyl ferulate (HF), and feruloylhexylamide (HFA). Cell proliferation, cell cycle progression, and apoptotic signaling were investigated in three human breast cancer cell lines, including estrogen-sensitive (MCF-7) and insensitive (MDA-MB-231 and HS578T). Furthermore, direct mitochondrial effects of parent and modified compounds were investigated by using isolated liver mitochondria. The results indicated that although the parent compounds presented no cytotoxicity, the new compounds inhibited cell proliferation and induced cell cycle alterations and cell death, with a predominant effect on MCF-7 cells. Interestingly, cell cycle data indicates that effects on nontumor BJ fibroblasts were predominantly cytostatic and not cytotoxic. The parent compounds and derivatives also promoted direct alterations on hepatic mitochondrial bioenergetics, although the most unexpected and never before reported one was that FA induces the mitochondrial permeability transition. The results show that the new caffeic and ferulic acid lipophilic derivatives show increased cytotoxicity toward human breast cancer cell lines, although the magnitude and type of effects appear to be dependent on the cell type. Mitochondrial data had no direct correspondence with effects on intact cells suggesting that this organelle may not be a critical component of the cellular effects observed. The data provide a rational approach to the design of effective cytotoxic lipophilic hydroxycinnamic derivatives that in the future could be profitably applied for chemopreventive and/or chemotherapeutic purposes.

 
 
3.  Impact of alkyl esters of caffeic and ferulic acids on tumor cell proliferation, cyclooxygenase enzyme, and lipid peroxidation.
Jayaprakasam B, Vanisree M, Zhang Y, Dewitt DL, Nair MG.
J Agric Food Chem. 2006 Jul 26;54(15):5375-81.
Source
Bioactive Natural Products and Phytoceuticals, Department of Horticulture and National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA.
Abstract
The antioxidant ferulic and caffeic acid phenolics are ubiquitous in plants and abundant in fruits and vegetables. We have synthesized a series of ferulic and caffeic acid esters and tested for tumor cell proliferation, cyclooxygenase enzymes (COX-1 and -2) and lipid peroxidation inhibitory activities in vitro. In the tumor cell proliferation assay, some of these esters showed excellent growth inhibition of colon cancer cells.


Thanks a lot
 

Kevin M. Folta said...

Ena, No problem. I'm home now so I can't grab them tonight, but tomorrow. Can you send me an email address? Thanks. Mine is kevinfolta at gmail

Ena Valikov said...

Hi Kevin.



Thanks for those articles. After scanning them my gut feeling is that phenolics aren’t it. I can imagine high levels in foods Slowing down growth of neoplasia, but I can’t imagine abnormally low levels independently of other xenobiotics Causing it.



If Seralini is correct in his threshold hypothesis –it needs to be either endocrine or epigenetic. I found a journal article you might find interesting in that it explains pretty well why your argument on lack of dose response falls apart in endocrinology: “Large effects from small exposures: nonlinear response of estrogen mimickers”: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1241550/pdf/ehp0111-000994.pdf

I just can’t shake TWO CP4 EPSPS supercharging metabolism of aromatics in NK603, one of which is a tad odd while turbo charged by the rice actin promoter.
The mechanism would seem to be either via CP4 EPSPS creating 1)a de-novo phytoestrogen not described in the literature so far, 2) or somehow catalyzing one of the steps leading to a more “virulent” estrogen (i.e. 4 hydroxyestradiol, 4-hydroxy catechol estrogen / estradiol quinones)

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC23537/

Or 3) enhancement of aromatase or

4) microRNAs.

Ena Valikov said...

Speaking of miRNAs, are the actual Monsanto transgenes sequences and the flanking sequences published? Is it possible to run a bioinformatics search for inverted repeats that can form hairpin RNAs upon read-through transcription on the transgene, the flanking regions paying attention to the anomalies listed below?

A band of -0.2 kb was detected in the “short run” test DNA (lane 5) indicating the presence of an additional fragment containing sequence from the P-racti/ractl intron. The -0.2-kb band was not detected in lane 2 because it was not retained on the gel after the “long run”. Further experiments which determined the sequence of the ends of the integrated DNA in NK603 revealed that an additional 217-bp fragment containing a portion of the enhancer region of the rice actin promoter was present in the reverse orientation proximal to the 3’ end of the transformation cassette, and that this small fragment maintained an EcoRV site 20-bp upstream from its 3’ end bordering corn genomic sequence (Figure 12). These findings confirmed and explained the results from this Southern blot analysis.The 217-bp fragment includes polylinker sequence (50 bp) and the first 167 bp of the enhancer region of the rice actin promoter, position -835 to -669 from the start of transcription as defined by McElroy et al. (1990). Neither the TATA box nor transcriptional initiation site is present within the fragment, which suggests that this fragment should not function as a promoter.

Nucleotide sequencing indicated that the sequence of the CP4 EPSPS gene within the second (3’ proximal) cassette differed by two nucleotides from the inserted sequence. This gave rise to a single amino acid substitution at position 214 of the expressed protein (leucine --> proline; variant protein referred to as: CP4 EPSPS L214P).
Analyses of specific PCR products from the 3’ terminus of the inserted DNA revealed that an additional segment comprising 305 bp of chloroplast DNA had been co-integrated. Bioinformatics analysis indicated that this sequence corresponded to a portion of the maize DNA-directed RNA polymerase alpha-subunit and ribosomal S11 protein. The source of this DNA was believed to be the chloroplast of the transformed cell
http://www.cera-gmc.org/?action=gm_crop_database&mode=ShowProd&data=NK603

Almost forgot. Given that this is a rebuttal of criticism of Seralini, and I hate double standards and asymmetry, I thought it would be appropriate to post the Hammond mirror piece for your criticism.
Results of a 13 week safety assurance study with rats fed grain from glyphosate tolerant corn by Monsanto Company, 800 N. Lindbergh, St Louis, MO, 63167, USA by Hammond et al.
http://cera-gmc.org/docs/articles/09-215-016.pdf
I notice it is published in that same low level rag: Food and Chemical Toxicology 42 (2004) 1003–1014, just exactly the same as Seralini. Does that mean that this safety study is rubbish as well, Kevin?

Ena Valikov said...

{And the food you eat is not sterile. You consume bacterial and fungal RNA every day by the bucket and none of that has ever been tested.}
I know that. You should see the look on people's faces when I show them mites that live on their eyelashes.
Moreover, the miRNA content of the blood has been well characterized—literally hundreds of papers.

{In all of these surveys why are dietary/bacteria/fungal miRNAs not found? I dunno.}
I believe it is because we have pancreatic ribonucleases, but I am not convinced they evolve as quickly as biotechnology in a span of a few decades.
http://www.ncbi.nlm.nih.gov/pubmed/16950759

Tiffany Villegas said...

Hello Kevin,

I wanted to start out by saying thank you for having this blog where everyone has the chance to pose their questions and opinions.

Both Seralini and Manuela Malatesta's tests showed that GMO foods induced negative effects on their subjects.

If one truly felt that these studies were inaccurate there would have been additional tests run to disprove the original findings. That is the way "good" science works.

I don't see that any additional testing has been done. This leads me to believe whole heartedly that the reason this was not done is because the belief is that the replicated studies would draw the same conclusion.

I wanted to know what your thoughts and opinions were on this question.

Why haven't the Seralini and Manuela Malatesta's tests ever replicated?

Could it be that in doing so one might prove that this is harmful and could potential be the cause of chronic disease and shorten people’s life spans.

Thank you,

Tiffany

Kevin M. Folta said...

2-part answer, part 1
Tiffany, thanks for the note. I appreciate your well thought out questions. Here are my thoughts:

Both Seralini and Manuela Malatesta's tests showed that GMO foods induced negative effects on their subjects.

But Seralini's work really didn't show that GMO foods induced negative effects. The criticisms are accurate and consistently show the sloppy science behind the study. I won't go into here, it has been brilliantly debunked by VIB in Belgium.

Malatesta's work is a little different. I think the researchers are certainly competent in the techniques, at least I don't know enough about them to be critical. What I do remember about these studies is that the diets were not reported, just that it was GMO and non-GMO. That leaves a lot of wiggle room in the soybean world. The test should be the transgenic and isogenic line. Soy is loaded with isoflavones, phytoestrogens, etc and they vary highly from line to line. That could more plausibly underlie the physiology reported.

If one truly felt that these studies were inaccurate there would have been additional tests run to disprove the original findings. That is the way "good" science works.

I'll disagree there. Good science never sets out to prove or disprove something. That's why Seralni's work is so awful. Good science tests a hypothesis. When we compare the tests that say there are no harmful effects and those that do, the ones that show harm come from the same labs and play fast and loose with the treatments. That's important. When we consider all of the work together the best studies outweigh sporadic reports of harm.

Plus, I'm not sure if the reported differences in Malatesta are indicative of deleterious health or just differences in cellular morphology. I'll revisit this.

I don't see that any additional testing has been done. This leads me to believe whole heartedly that the reason this was not done is because the belief is that the replicated studies would draw the same conclusion.

I think you have it backwards. Replicated studies with statistically meaningful numbers likely showed no treatment effect and were not published. Remember, the scientist that shows definitive proof that 95% of USA acreage is poison will receive a Nobel Prize. I hope it's me!

If I wanted to do this study and wrote to NIH or NSF for funding they would never fund it. There's no plausible mechanism for harm beyond mundane explanations. It would never be replicated, because the primary findings are not trustworthy.

Kevin M. Folta said...

Two part answer, part 2

I wanted to know what your thoughts and opinions were on this question.

Why haven't the Seralini and Manuela Malatesta's tests ever replicated?

Could it be that in doing so one might prove that this is harmful and could potential be the cause of chronic disease and shorten people’s life spans.


Again, if it is real, it should be reproducible. Other labs, many others, should jump all over this to be the first to show that 95% of USA, Brazil and Argentina's acreage is toxic in a definitive, solid study. If these first papers held water you'd see a mad dash for more studies, bigger studies.

But real scientists look at this report and see no reason to follow up. It is scientifically DOA.

And there is a little double standard at play when the opponents of transgenic technology cry foul when an author on a paper has some ancient tenuous tie to Monsanto, yet Seralini's work is funded by an anti-GMO organization and he heads an institute designed to find fault with GMO, and he publishes opinion books about GMO harm.

Tiffany, you, me, Ena, and the rest are all on the same page. We want more safe food that is better for people and the environment. Biotech has its strengths and limits. However, we can't let sloppy science shape our decisions. If it is harmful, then that's fine, I'll freely admit it and endorse the work. However, it is going to take some good experiments with solid numbers to exceed the evidence threshold needed to be convincing. If it is real, then it would be do-able.

Ena Valikov said...

An article you might find interesting

A novel endocrine-disrupting agent in corn with mitogenic activity in human breast and prostatic cancer cells.
Barry Markaverich, Shaila Mani, Mary Ann Alejandro, Andrea Mitchell, David Markaverich, Trellis Brown, Claudia Velez-Trippe, Chris Murchison, Bert O'Malley, and Robert Faith
Author information ► Copyright and License information ►
See letter "Corn and corn-derived products: sources of endocrine disruptors." in volume 111 on page A691.
This article has been cited by other articles in PMC.
Abstract
Housing adult rats on ground corncob bedding impedes male and female mating behavior and causes acyclicity in females. The suppressive effects on ovarian cyclicity are mimicked by a mitogenic agent purified from the ground corncob bedding material (corn mitogen; CM), which stimulates the proliferation of estrogen receptor (ER)-positive (MCF-7 cells) and ER-negative (MDA-MD-231 cells) breast cancer cells. Purified CM does not compete for [(3)H]estradiol binding to ER or nuclear type II sites, and its effects on MCF-7 breast cancer cell proliferation are not blocked by the antiestrogen ICI-182,780. These results suggest that the active component is unlikely to be a phytoestrogen, bioflavonoid, mycotoxin, or other known endocrine-disrupting agent that modifies cell growth via ER or type II [(3)H]estradiol binding sites. CM also stimulates the proliferation of PC-3 human prostatic cancer cells in vitro, and the growth rate of PC-3 cell xenografts is accelerated in nude male mice housed on ground corncob as opposed to pure cellulose bedding. Consequently, this endocrine-disrupting agent in ground corncob bedding may influence behavioral and physiologic reproductive response profiles and malignant cell proliferation in experimental animals. Fresh corn (kernels and cob) or corn tortillas also contain CM, indicating that human exposure is likely; consequently, CM and/or related mitogens in corn products may influence human health and development.

Full Text
The Full Text of this article is available as a PDF (559K).

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1240732/

Cheers,
Ena

Kevin M. Folta said...

Didn't find it that remarkable. Seems like okay data and has its limitations for interpretation.

We've known for a long time that you don't house rodents on cedar chips or other wood that emits volatile hydrocarbons,etc. They don't say anything about the genotype of the corn or even who made it.

The Discussion suggests that corn itself may maintain CM as it is found in tortillas.

So based on these data from a limited study with some nice dose-response data from multiple models, to you recommend to not eat corn?

Ena Valikov said...

Given that the corn in the taco shells and the bedding was unlabeled, it is obviously impossible to tell which variety of corn;though it is likely it was transgenic. It seems a good idea to have the manufacturers of transgenic corn seeds analyze their variety for leukotoxin diols....

Leukotoxin diols from ground corncob bedding disrupt estrous cyclicity in rats and stimulate MCF-7 breast cancer cell proliferation.
Markaverich BM, Crowley JR, Alejandro MA, Shoulars K, Casajuna N, Mani S, Reyna A, Sharp J.
Source
Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA. barrym@bcm.tmc.edu
Abstract
Previous studies in our laboratory demonstrated that high-performance liquid chromatography (HPLC) analysis of ground corncob bedding extracts characterized two components (peak I and peak II) that disrupted endocrine function in male and female rats and stimulated breast and prostate cancer cell proliferation in vitro and in vivo. The active substances in peak I were identified as an isomeric mixture of 9,12-oxy-10,13-dihydroxyoctadecanoic acid and 10,13-oxy-9,12-dihydroxyoctadecanoic acid, collectively designated tetrahydrofurandiols (THF-diols). Studies presented here describe the purification and identification of the HPLC peak II component as 9,10-dihydroxy-12-octadecenoic acid (leukotoxin diol; LTX-diol), a well-known leukotoxin. A synthetic mixture of LTX-diol and 12,13-dihydroxy-9-octadecenoic acid (iso-leukotoxin diol; i-LTX-diol) isomers was separated by HPLC, and each isomer stimulated (p < 0.001) MCF-7 cell proliferation in an equivalent fashion. The LTX-diol isomers failed to compete for [3H]estradiol binding to the estrogen receptor or nuclear type II sites, even though oral administration of very low doses of these compounds (>> 0.8 mg/kg body weight/day) disrupted estrous cyclicity in female rats. The LTX-diols did not disrupt male sexual behavior, suggesting that sex differences exist in response to these endocrine-disruptive agents. http://www.ncbi.nlm.nih.gov/pubmed/16330350