Thursday, April 28, 2011

Let Us Honor Ancel Keys, Our Patron, As We Cherry Pick Studies to Bash Fructose (Revised and Extended)

My apologies to anyone who received this in their RSS feed on Thursday as a teaser.  I have now revised and extended it to include several studies showing that diet-induced obesity can be achieved in rats and mice without using any sugar at all, and have included a clearer conclusion.

Ancel Keys is best known in the assorted real food communities, or at least the assortment of real food communities friendly to dietary fat, for his infamous cherry picking.

Keys had presented data from six countries, purporting to show a clear linear relationship between the amount of fat consumed in a country and its incidence of heart disease.  This graph is shown on the left below.  The one problem was that data was available for 22 countries at the time, and including that data demolished the relationship.  The "true" graph is shown below on the right.

December 22, 2011 Update:  It would be more appropriate to say that including the data "greatly diminished" the relationship than that it "demolished" the relationship because the positive relationship still exists.  See this excellent analysis by Denise Minger.

If we are going to criticize Keys for cherry picking, it behooves us to root out cherry picking from our own communities as well.  This requires constant self-criticism, because bias is human nature, and anyone who isn't engaged in a devoted battle to overcome their own bias will be its prisoner.  I myself cannot claim to always be victorious in this battle, but I do try.

I consider Dr. Robert Lustig an ally in the fight for real food.  He makes as his primary enemies junk foods rich in refined sugar and high fructose corn syrup, and I agree these should be targeted as likely the most important causes of chronic disease.  Dr. Lustig and many in the blogosphere, however, are circulating the claim that high-fat diets only induce obesity in laboratory animals if they are also high in sucrose.  This simply isn't true.

Let's take a look at these diets.

Thursday, April 21, 2011

New Fatty Liver Study Shows that Carbohydrate Restriction Causes Statistical Anomalies

by Chris Masterjohn

A new study claims to show that carbohydrate restriction is superior to calorie restriction at improving fatty liver disease:

Browning JD, Baker JA, Rogers T, Davis J, Satapati S, Burgess SC.  Short-term weight loss and hepatic triglyceride reduction: evidence of a metabolic advantage with dietary carbohydrate restriction.  Am J Clin Nutr. 2011

In this study the authors randomly assigned 18 people to spend two weeks consuming either a low-calorie diet (~1325 kcal/day) or a low-carb, (<20 g/day), high-protein (33% kcal) diet. The low-carb, high-protein diet led to a greater reduction in the amount of fat stored in the liver.

The authors begin their manuscript by stating as a matter of fact that insulin resistance causes fatty liver, without even acknowledging the competing hypothesis that fatty liver causes insulin resistance. 
After the holidays, I'll provide a more comprehensive post addressing whether it makes any sense to single out "carbohydrates" as causing fatty liver (hint: it doesn't), but for now let's take a brief look at this study.

Sunday, April 17, 2011

The New Genetics -- Part V: Is the Intestinal Microbiome Part of Our Genome?

Has the human genome project really been completed?  One could argue that in fact it will not be completed until its sequel, the Human Microbiome Project, is completed.  One set of authors referred to it as "another phase of the 'human’ genome sequencing project."

The available evidence suggests that intestinal microbes are inherited from parent to offspring, just like the biologically heritable components (not all of which are genetic) of our Homo sapiens cells are.  Just like the heritable components of our H. sapiens cells, these microbes appear to substantially contribute to our phenotype — that is, the overall end result of who we are.  

What is particularly fascinating, however, is that there is evidence that variations in our metabolism that could be induced by diet or environment, such as leptin signaling, can alter the relative abundance of certain gut microbes.  Thus, not only do these microbes drive our phenotype, but our phenotype drives the microbes!   

This highlights a huge problem of inferring cause-and-effect relationships into inheritance patterns.  While traditional "genetic" inheritance can be inferred in certain cases from solid mathematical patterns, the mere fact that a trait is heritable not only does not show that it is "genetic" but it does not even show that the inheritance is due to our H. sapiens cells, nor does it show that the trait cannot be changed by normalizing our metabolism through dietary, lifestyle, and environmental modifications.

Saturday, April 16, 2011

What Grad School and Professional Research Should Be Like

As I was traveling to Experimental Biology this past weekend with my lab mate and her boyfriend, who is a doctoral student in another health-related department, I shared my thoughts on what grad school should be like.  Naturally, the reaction was a laugh, and a "ha, that would be awesome."

Here's my idea.

As researchers in nutrition or exercise science or other health-related fields, we should first and foremost be setting examples of how to live a healthy life to the general population.  That means the priorities should be aligned as follows:

Friday, April 15, 2011

Highlights From Experimental Biology 2011

by Chris Masterjohn

I arrived in Washington, DC on Friday evening to meet with 13,000 other biological scientists for the Experimental Biology 2011 meeting and just got home Wednesday night.  

I was there to give a 15-minute presentation on how I fed an overdose of fructose to a couple dozen rats and it didn't do any of the nasty things I expected it to.  You'll be hearing lots more about fructose from me in the near future as I try to figure out what happened.

One of the coolest presentations I saw was a talk Peter Friedl of the German Nijmegen Center for Molecular Life Sciences gave entitled "Dynamic imaging of cancer invasion, plasticity, and resistance."  This was the first talk I saw of the conference, on Saturday morning.  Friedl showed textbook images of cancer metastasis, which show isolated cells dropping off from a cancer and then making their lonely but merry way to invade other tissues.  Then he showed videos proving it was false.

His remarkable live streaming images showed that cancers actually send forth highly organized finger-like projections like these:

Those bright strands  are highly organized projections made up of individual and continuously dividing cells.  A minority of their expansion is due to the fact that they are traveling forward, led by a small cluster of a few cells or sometimes an individual cell at the tip, while the majority of their growth comes from the fact that they are actively dividing, pushing forward as they do so.

Sunday I gave my own talk and later I was student chair of the fourth Dietary Bioactive Components session, focusing on antioxidant and anti-inflammatory compounds.  Little did I realize that the job of the student chair would be to manage the twitter feed.

And of course there was way too much stuff going on at this conference for anyone to possibly see it all, so check out the #eb2011 hashtag on twitter to find other tweets and blog posts, or maybe search Google.

The best talk I saw was on Monday.  Paul Kubes of the University of Calgary gave a talk entitled "Sterile versus infectious inflammation... an identical immune response?"  Kubes has a hilarious sense of humor, which livened up the talk a bit, but the content was simply astounding.  In his presentation he showed that immune cells called neutrophils respond to bacterial infection by spilling out their DNA to form webs that catch bacteria.  

Here are some neutrophils:

When they are exposed to Staphylococcus aureus, they start spilling out their DNA, shown in green:

Over time, these nets greatly expand:

Through electron microscopy we can see that these DNA-nets form lattice-like webs that catch Staph aureus just like spider webs catch insects:

Other images show that these nets protrude out of a small bubble on the surface of the neutrophil's membrane, kind of like how Spider Man shoots webs out of his wrist.

What was particularly amazing was watching live streaming video of this process, where the denucleated neutrophils could be seen crawling around eating the bacteria they had caught in their DNA-webs.  

Conclusion?  Neutrophils are kind of like spiders.  Err, I guess they are kind of superheroes so maybe they are more like Spider Man.

And man, with all these live streaming photos of fluorescing molecules, who wouldn't want to get into this line of work?  Oh right, I almost forgot.  For a few hundred reasons why no one in their right mind would get into this line of work, check out PhD Comics.

Although, this Lady Gaga rendition sums it up rather succintly:

On Tuesday, I went to a special session put on by the Institute of Medicine's Food and Nutrition Board (FNB) about the new DRIs for calcium and vitamin D.  They focused mostly on vitamin D.  Presenters included the chair of the FNB, Dennis Bier, the director of the FNB, Linda Meyers,  the chair of the committee that wrote the vitamin D report, Catherine Ross, and several other people involved in writing the vitamin D report such as Christine Taylor, Patsy Brannon, and Susan Mayne.

Mayne, a cancer epidemiologist from Yale, discussed a list of what she considered myths floating around the internet and elsewhere about the vitamin D report:

You can click the picture to enlarge it.

And here are the facts, as she saw them:

I apologize the photos are kind of blurry.

Regardless of whether the IOM's FNB got the best estimate of the vitamin D requirement exactly right, I agreed with the gist of Payne's talk.  I got the impression as soon as I started reading the 999 page report that there was an awful lot of misunderstanding about the report and its purpose.

When I'm done reading it, I'll post my analysis on my other blog, Mother Nature Obeyed.

Unfortunately, I missed the Wednesday session entitled, "Exploring the Factors That Impact Blood Cholesterol and Heart Disease Risk: Is Dietary Cholesterol as Bad For You as History Leads Us to Believe?"  The director of our graduate program here at UConn, gave a talk entitled, "Rethinking Dietary Cholesterol: A Critical Review of Existing Lieterature."

She told me the moderated question and answer session produced quite a lively debate.

The best part of the conference was meeting new friends.  I met nutrition journalist David Despain on Saturday night.  Here's a picture of Melissa, David, and me parting at the subway after chowing down on some delicious Mexican food together:

Without love and friendship, science would be pretty boring.

Read more about the author, Chris Masterjohn, PhD, here.

Wednesday, April 13, 2011

Does Dietary Choline Contribute to Heart Disease?

by Chris Masterjohn

New blog over at Mother Nature Obeyed on the WAPF site:

Does Dietary Choline Contribute to Heart Disease?

Read more about the author, Chris Masterjohn, PhD, here.

Thursday, April 7, 2011

My New Wise Traditions Article on Nonalcoholic Fatty Liver Disease

by Chris Masterjohn

I have a new article on nonalcoholic fatty liver disease appearing in the outgoing issue of Wise Traditions, and the online version just went up on the web site:

The article contains a lot of information that I've already posted on this blog, but it also contains quite a bit of additional information, and pulls everything together a little more succinctly.

In the article, I argue that fatty liver is essentially the consequence of several independent nutritional assaults.  These include 1) nutrient-poor refined foods, 2) choline deficiency, and 3) polyunsaturated oils.  Fatty liver can basically be divided into two stages, the first involving the accumulation of fat, and the second involving the progression of inflammation.  The first, I argue, results from too much caloric energy entering the liver without sufficient nutrition to process it, while the second results from any type of oxidative and inflammatory insults, with excess dietary linoleic acid making a special contribution.

I point out that none of the animal models, whether they are high in fructose, high in fat, or deficient in choline, actually look like human fatty liver until they are combined together, emphasizing that human fatty liver is likely a result of a combination of insults.

I also point out that the liver plays an enormous role in our overall health, from regulating blood sugar and blood lipids to filtering bacteria and their toxins to regulating hormones and vitamin D, and that fatty liver is a much more powerful predictor of heart disease risk even than "metabolic syndrome."

For those of you willing to support the Weston A. Price Foundation, I would highly recommend subscribing to the journal.  My articles contain a lot of sidebars, and there is really no pleasant way to arrange them online.  Reading the article in print is much more enjoyable.  Nevertheless, I believe most of you would find the information valuable, so I invite you to hop on over and read the article online.

Read more about the author, Chris Masterjohn, PhD, here.

Wednesday, April 6, 2011

What No One Is Saying About Zonulin -- Is Celiac About More Than Genes and Gluten?

by Chris Masterjohn

In my last gluten post, I discussed why the ex vivo results of Dr. Fasano's 2006 study cannot logically be construed to show that gluten causes leaky gut in people without celiac disease, and why the available evidence suggests that people considered to have non-celiac gluten sensitivity do not have leaky gut.

Nevertheless, this study is a goldmine of valuable ideas that no one is touching.

Not all of the study's results should be considered ex vivo.  They took intestinal tissue from celiacs in remission and from non-celiac controls with digestive complaints, and measured the amount of zonulin in those tissues before performing any experiments on them.

Remarkably, they found that celiacs produce 30 times as much zonulin as non-celiacs, even though the non-celiacs were not eating gluten-free diets while the celiacs had been off gluten for over two years!

Here's a graph of their data:

This is remarkable because even though the point of the study was to show that gluten increases zonulin production, the controls were eating gluten yet had infinitesimal levels of zonulin production, while the celiacs had not eaten gluten for at least two years yet still had very high levels of zonulin production.  This suggests that something besides gluten may be causing zonulin production in celiacs.

They found similar, though less dramatic, results for intestinal permeability:

Here they measured trans-epithelial electrical resistance (TEER) of intestinal tissue taken from gluten-free celiacs and gluten-eating controls.  TEER is an estimation of the leakiness of the gut, where a lower value indicates a greater level of leakiness or permeability.  They found that tissues taken from controls who had been eating gluten had three-fold less leakiness compared to celiacs who had been off gluten for over two years.  This, again, suggests that something besides gluten may be contributing to leaky gut in people with celiac.

What is causing the persistently elevated zonulin in celiacs, or the somewhat less severe persistent elevation in gut permeability?  It could just be that these subjects need to adhere to a gluten-free diet more strictly or for much longer than two years to fully resolve these issues.  Or, it may be that certain types of intestinal dysbiosis (improper balance of bacteria and yeasts in the intestines) prime genetically susceptible individuals to develop celiac in response to gluten.

Dr. Fasano's group has also published a study showing that bacteria such as E. coli and Salmonella stimulate zonulin production in isolated intestinal tissue, and another recent study showed that short-term inoculation of rats with E. coli and Shigella enhanced the ability of gluten to cause intestinal damage while inoculation with Bifidus bacteria virtually eliminated gluten's ability to cause damage.  Neither of these studies show that dysbiosis contributes to celiac, or that it is responsible for the persistence of zonulin production on a gluten-free diet, but they offer strong support to the plausibility of these hypotheses.

In any case, it is true that Dr. Fasano's 2006 study showed that gluten was capable of increasing zonulin and consequently increasing gut permeability ex vivo in intestinal tissue from both celiac and non-celiac subjects.  Still, the effect in tissue taken from non-celiac individuals is pretty small.

Here's the effect on zonulin:

Here they show that in the mucosal layer, but not the serosal layer, zonulin increases in response to gluten ex vivo, but that zonulin concentrations are dramatically higher in celiacs to begin with and remain dramatically lower in non-celiacs at all time points.  

The mucosal layer is more superficial whereas the serosal layer is more deep.

Similar results were also seen for TEER, their estimation of gut integrity:

Here again we see that although gluten decreased gut integrity even in tissue isolated from subjects without celiac who were eating gluten as part of their normal diet, it never declined even close to the level seen in celiacs who had been gluten-free for two years.

As I pointed out in my last gluten post, these ex vivo experimental results showing that gluten proteins increase zonulin and leaky gut in isolated intestinal tissues cannot logically be construed as evidence that gluten causes leaky gut in live humans without celiac disease.  There are far too many factors that could intervene in a live human to change the result.  And indeed, Dr. Fasano's most recent study showing that people with non-celiac gluten sensitivity do not have leaky gut and the Australian study showing that gluten does not cause leaky gut in such individuals directly refute this concept. 

Whether gluten contributes to the leaky gut seen in other diseases such as autism remains to be seen, but based on this data, one could could suggest that this is the case as a plausible hypothesis.

Ultimately, however, the most remarkable finding of this study is the massive persistence of zonulin production and leaky gut in celiacs even after they have been gluten-free for two years.  The authors noted this in their conclusion:
Nevertheless, zonulin is markedly up-regulated in subjects affected by [celiac disease], even when treated with a gluten-free diet.  This up-regulation is associated with increased baseline gut permeability, and an increased amplitude and duration of gluten-induced zonulin release when compared with non-[celiac disease] intestinal samples.  Despite the presence of measurable zonulin response in both [celiac disease] and non-[celiac disease] subjects, [celiac disease] patients appear to reach a critical threshold of intestinal permeability upon gliadin exposure that is not reached in non-[celiac disease] intestinal mucosa.
This is remarkable because it suggests that celiac is about more than just genes and gluten.  I will revisit this topic when I get my food toxins and food intolerances series going.  In the mean time, I owe you all a sequel to my last LDL post, and  then it'll be back to fructose for a while.

Read more about the author, Chris Masterjohn, PhD, here.

Monday, April 4, 2011

How to Properly Interpret Ex Vivo Studies -- Gluten and Leaky Gut As an Example

by Chris Masterjohn

There seems to be some confusion about how to appropriately interpret ex vivo studies, which are studies that are not conducted in a living organism. 

Worse than that, there appears to be a common and rather dramatically misleading presentation of the data in Dr. Allesio Fasano's excellent study linking gluten to zonulin production.  But I'll cover that in my next gluten post.

In any case, some people construe this study as showing that eating wheat or gluten causes leaky gut in people who do not have celiac, when in actuality the authors treated isolated tissues and tissue models with gluten rather than feeding gluten to people and seeing what happened.

A study conducted in isolated human cells or tissues is not a "human study."  A study conducted in isolated human cells or tissues is useful for three things:
  • generating the hypothesis that the effect also occurs in live humans.
  • providing a possible explanation for existing data obtained from live humans. 
  • characterizing the mechanistic details of the molecular biology of some phenomenon that would be impossibly invasive to characterize in living humans.

Whatever such a study is, it is not a test of the hypothesis that the phenomenon occurs in humans.

Saturday, April 2, 2011

Is Gluten Making My Gut Leaky? (A Shorter Post!)

by Chris Masterjohn

While some people really liked my last blog post, one commenter asked me to writer a shorter post next time.  So here's a shorter post.

A commenter on Paul Jaminet's blog recently suggested that the reason I developed some arthritic symptoms including a limp during a brief stint eating two to three sweet potatoes per day was not because they are extremely high in oxalates, which was my guess, but because wheat is giving me leaky gut, allowing the lectins in sweet potatoes (and all other plant foods, he suggests) to cause an autoimmune reaction.

I actually don't consume a whole lot of wheat and most of my gluten comes from rye that has been soured for 24 hours, which is likely to be pretty low in gluten. 

However, I believe I had been gluten-free for over a year before the sweet potato incident.  In looking back at past emails and posts on various lists, I find that I went gluten-free and casein-free back in July of 2005 and that the sweet potato incident happened mid-October, 2006.  Although I do not have any record of the exact date I reintroduced the 24-hour soured rye bread, the earliest record I can find is a post I wrote on November 23, 2006 saying I reintroduced it "recently."

Although my memory of what I ate four to five years ago is not perfect, I believe the reason I was eating so many sweet potatoes to begin with was because I was trying to do the FAILSAFE diet while also remaining gluten-free and casein-free, and I was finding it difficult to obtain sufficient calories without using lots of vegetables or starches as a vehicle for fat.  So I feel pretty confident I was gluten-free at the time.

In any case, given the two recent studies showing that gluten and wheat do not cause leaky gut in people without celiac, I'm not sure why anyone would propose that I would have a leaky gut from eating wheat, even if I were actually eating a substantial amount of wheat.

As I pointed out in my last gluten post, a recent paper by gluten researcher Dr. Allesio Fasano and colleagues found that intestinal permeability was even lower in people they judged as having non-celiac gluten sensitivity who had been eating gluten under supervision for four months when they compared them to control subjects, whereas those with active celiac disease had greater intestinal permeability.

Likewise, the recent double-blind, placebo-controlled gluten trial  found that six weeks of gluten consumption did not cause any change in intestinal permeability in subjects the authors judged as having non-celiac gluten sensitivity.

I realize these studies are new and these findings may not be well known, but it seems that it is time to stop assuming that gluten causes intestinal permeability in anyone except people with celiac, unless some contrary information emerges.  It is, of course, possible that some subset of people without celiac exists in whom this occurs, but one would think that if it occurs in anyone without celiac, it would be in people considered to be gluten-sensitive.

If anyone knows of clear evidence that gluten alters intestinal permeability in humans without celiac, please post a link to the study in the comments.

Read more about the author, Chris Masterjohn, PhD, here.

Friday, April 1, 2011

How a Study Can Show Something to Be True When It's Completely False -- Regression to the Mean

by Chris Masterjohn

In a previous post, "The Great Unknown: Using the Statistics to Explore the Secret Depths of Unpublished Research," I discussed one way a study can show something to be true when it's false, or vice versa.  

If some nutrient or drug has a "true" biological effect, and we repeat many studies of the phenomenon, we would expect a handful of them to show a much greater effect than the real one and a handful of them to show a much lesser effect.  In some instances, we might even expect to see the opposite effect turn up by random chance in a few studies.  And if some nutrient or drug has no effect whatsoever, the bias against publishing negative results might leave us with a small handful of conflicting studies, some showing a positive effect and some showing a negative effect, but few if any showing the truth  — no effect.

In my most recent post on gluten, I mentioned yet another way a study can show something to be true when it's false: regression to the mean.  I also discussed regression to the mean in my 2006 article "Myth: One High Saturated Fat Meal Can Be Bad", which was the basis for a much shorter letter I published in the Journal of the American College of Cardiology.

In this post I'll explain how this phenomenon can easily cause fake findings to be paraded around as true, and then use the real-life example of this ridiculous study making the absurd claim that statins cure vitamin D deficiency.