 rough so I'm not going to be offended if there's a large rate of attrition but I don't feel so bad after seeing Robert Lustig's talk that was given at an academic level. So for those of you who don't know me I'm an organic chemist I obtained my PhD in organic chemistry from Harvard University under the guidance of professor Eric Jacobson doing organo catalysis. If you're into scrabble those are all great words to use. And I have been on the road giving nutrition talks for about two years now and I have Rob Wolf to thank for that for ruining my life. There came a time when I had a choice to make and that choice was to either join the pharmaceutical industry or use my knowledge of nutrition to help people and I chose the latter. So thanks. This talk came about when I asked myself the question you know if I had to present this material right here this is the department's the CCB the Department of Chemistry and Chemical Biology at Harvard University and right down here is Pfizer lecture hall and Pfizer lecture hall is where all of the seminars are given and I thought to myself how would I present this material if I had to present it in front of my peers in the chemistry department and really there's no shenanigans to be made you can't make any exaggerations. I have of course participated in witness many chemistry talks there and I can tell you right now that people who make overstate their claims or make mistakes are treated to a question-and-answer period that makes a CIA interrogation look like a teeny-bopper interview. So this is the talk that you're about to see is essentially what came out of this thought process. So here's how I would not present this information. I would not present this information in the way that it is currently presented in the blogosphere and that is that our ancestors and modern hunter-gatherers consume the diet mostly devoid of grains, legumes, and dairy. Our ancestors and modern hunter-gatherers were virtually free of diseases of civilization and of course the invalid inference would be that consuming a diet mostly devoid of grains, legumes, and dairy will allow us to be free of disease of civilization and the reason why that isn't valid is because this material is only observational in nature. It does not establish cause and effect it only gives you correlation and I see this mistake made in the blogosphere constantly in that people start off with these observations and they think that they do establish cause and effect and they don't. I see a lot of people poo-poo on Ansel Keys' work well guess what you could you know do the same with this. So Boyd Eaton himself recently said so in a paper that he published he says such evidence could only suggest testable hypotheses and that recommendations must ultimately rest on more conventional epidemiological clinical and laboratory studies and that's absolutely correct. Now I'm not saying that this is that this material is useless. Observational epidemiology is very useful for generating hypotheses. It's very useful for asking questions. So here's a graph that was published in the paleo diet update by Professor Cordane and his team and you can see on the left the typical Western diet and the fat ratios and then the same ratios and actually the average ratios of various hunter-gatherer diets and someone looking at this graph could think well is saturated fat responsible for the poor health outcome of the Western diet because you can see there's much more on this side than on that side but you could also ask hey maybe saturated fat has been official and the hunter-gatherers weren't getting enough. You can't answer that question with this graph. You can only ask that question. You could also ask well I wonder if the levels of polyunsaturated fatty acids of those hunter-gatherers were too high. Maybe the ones of you know today's population are too low. You don't know you can only ask the question. The invalid inference of course and one that I hear a lot is that what you really want to do is emulate the fatty acid ratio of the hunter-gatherers if what you're looking for is optimal health. You cannot make that statement based on these observations. Absolutely not. Here is another graph. This one comes from the paleo diet for athletes by Freel and Cordane and looking at these graphs you could ask the questions are Americans consuming too many calories from carbohydrate? Maybe maybe not. Are Americans not consuming enough protein? Maybe we don't know. The invalid inference of course would be that the calculated average macronutrient ratio of hunter-gatherer populations is the healthiest. If you make statements such as these in front of a core science audience based on observational epidemiology no one will take you seriously. And my colleagues will not take me seriously. This is a book that I've recently been through and I'd like to tell you I'd like to read a quote from the book because I see a lot of people making this mistake and that everybody should be in this particular macronutrient ratio. Anyone who comes along and says everyone should be 30, 30, 40. Everyone should be 20, 20, 60. No one in the core sciences is going to take you seriously and here's why. The immense biological variability among individuals has overpowering significance. The examples are shown above and support to this thesis. Much theoretical and practical knowledge can be gained from study of biological individuality. Nutritional variability on an international level requires that we be aware of these intricacies and interactions. The global approach necessitates caution and implementing world-wide nutritional programs based on mean requirements derived in a specific culture. This narrow orientation overlooks the importance of man's nutritional individuality, a reflection of evolution. This is an excellent book which makes the case that food was a huge driving force in evolution and that many of us are different as a result. A few examples that have been brought up of course is the human evolution or adaptation to lactase persistence. Another one is for amylase. Some of us are better adapted to starch diets, for example. Again, someone who comes along and says everyone should be on a low carb diet, I highly disagree. I'm sorry. It depends on the context. As Gary stated yesterday, you know what? Someone who has metabolic syndrome, you've got impaired blood sugar control, you probably should be on a low carb diet. Does that mean low carb for life? Probably not. It depends on how much beta cell damage was done. Does that say anything about food quality? No, it doesn't. Maybe you should focus on that too. This is one of my favorites and I'll end with a paleobashing on this one. I hear this argument all the time. We evolved over millions of millions of years without consuming the foods that became readily available only after the advent of agriculture, hence we are not adapted to them. And if you were to go to a biology or an evolution department and you were to make this statement, you would get smacked really hard. Because there's some questions that are left unanswered. Is it really safe to assume that we are not adapted to any food that became readily available and part of the human diet after the advent of agriculture? And of course, doesn't this suggest that a species cannot discover a new, better source of food? I mean, I'll give you that. After the advent of agriculture, our record has been pretty darn bad. But the possibility is still there that we've discovered something a little better than what was there before. And it still doesn't mean that what was there before is really any better than something that we could discover afterwards. For example, should I be consuming large quantities of seeds if they're sunflower seeds? Just because they're quote unquote paleo. So this is what I state right here. So these are, all of these are invalid inferences, and you should not use them when you're trying to convince people that they should go, or they should follow this lifestyle. So how, actually I'm not done, there's one more. At the end of the day, you have to remember that all hypotheses are valid. It doesn't matter where they come from. They all have to be tested before they're shown to be invalid or not valid. So they're all worth considering. Just because your hypothesis relies on quote unquote evolution doesn't make you any more right than anyone else. Okay, and if you go into the literature, if you ask a question, look at the evolution, think that you're right, and then go into the literature and look for papers that support your claims, then you're doing a selection bias. And this is poor science. Alright, so approaching things this way is only going to lead to poor science. No one in the core sciences is going to take you seriously. And by the way, what I mean by core sciences is mathematics, physics, chemistry, and biology in that order. Biology is the chemistry of living organisms. Chemistry is the interaction, the physical interactions of atoms. Physics is described by mathematics. So how would I, you know, approach this in front of a chemistry audience? Well, of course I'd approach it with chemistry. And I'm going to attempt to try to teach you a little bit of chemistry real quick. I'm not going to go through the entire table. I'm just going to go through the first two rows. So a little bit about valence and bonding. I'm sure some of you are going to want to run out of here screaming. But if you're counting the columns in the periodic table, this is giving you an indication of how many valence electrons are around the atom and how many bonds that atom can form. So hydrogen's in the first one. Well, guess what? It forms only one bond. Here's the hydrogen molecule and it's linear. Carbon is in one, two, three, four. Don't be fooled by the number. So carbon can form four bonds and those four bonds are orient, their orientation around that carbon atom is what we call tetrahedral. So imagine that carbon is sitting right in the center and then the hydrogen atoms are at the apex of the tetrahedron. Ammonia and water, they actually form three and two bonds respectively. Even though when you count one, two, three, four, five, you think nitrogen would four, five bonds, but that add, the electrons like to pair up. So you have one electron pair and three bonds. With oxygen, you've got six, but two electron pairs, two bonds. And then with something like fluorine, you have six electron pairs and it only forms one bond. These are distorted tetrahedrons because those lone pairs take up a lot of space. So now you can start drawing hydrocarbons. So here's methane. I've shown already the simplest. If you hook up two of these, you've got ethane. Note that although the bond angle is technically in a tetrahedron 109.5, we draw these in a 120-degree angle because it makes it simpler and nicer to draw. But you can realize I'm going from methane, ethane, propane, butane, pentane, and the pent means five, you know, butte four, probe three, octadecane, eighteen. It's a chore to start drawing these things. It really is a chore. I don't get to that in a minute. So I just want to cover a little bit about double bond geometry. You've got two flavors here, two garden varieties, trans and cis. Trans means that when you have a double bond, note that carbon always makes four bonds, right? So here it still has one, two, three, four because of that double bond. Here it has one, two, three, four because of four single bonds. If the hydrogen atoms are on opposite sides of the double bond, we say they're trans. If they're on the same side of the double bond, we say that they're cis. So here we have elatic acid, which is the major monounsaturated fatty acid created during the hydrogenation of polyunsaturated fatty acids. And then here's oleic acid, which you'll find in olive oil and nuts and avocados. And you'll notice that in one molecule the double bond is trans and then in the other, the double bond is cis. Now chemists would call these compounds this way because it actually gives you information about the structure, whereas a lot of people like to just give them names and that says absolutely nothing about the structure or the double bond geometry. So here's how we simplify things because these things get really ugly really quickly. We actually delete all of the hydrogen atoms and just, you know, they're implied. So if you've got, and actually the carbons are at the connection of all lines, so there's carbon atom here, here and here, and at the end of all lines. So if there's a carbon atom here and it only has one bond, that means there has to be three hydrogen atoms around it. So we delete everything, the carbons aren't drawn, hydrogen aren't drawn, everything is implied. The end carbon, you can actually draw it if you want a CH3 and you can abbreviate it as methyl if you want. And then the double bonds are drawn along. So whenever you look something up on Wikipedia from now on and you see these structures, hopefully they will mean a little bit more to you. Maybe not, maybe I failed miserably. Before I go on, just a little bit about stereochemistry. It turns out that if you have four different substituents on carbon, you introduce something called stereochemistry. So if I have dichlorofluoromethane and some of you might be familiar with these molecules, these are HFCs, they're bad for the ozone layer, you take this molecule and you make its mirror image, you get this molecule and then if you flip it 180 degrees, you can see that the molecules are identical. These are superimposable mirror images. However, if I have four different substituents, I grab the mirror image, I flip it 180 degrees because I want to overlay them, they are no longer superimposable. And this is the same thing between your left hand and your right hand. They're not superimposable, they're mirror images of one another. So in this case we have something called stereochemistry and these two molecules are different. I'm not going to go into the nomenclature but we would call this one S and we would call this one R. So why is this important? Well, the talk is all about credibility. How do you present this material while retaining as much credibility as possible? And really there's no God without believers, there's no researcher without credibility. And that's how it goes. So I just recently read this article and let me read this. Synthroid, which is level thyroxine sodium, as the name implies is not natural, it's not bioequivalent to the hormone thyroxine and it's produced by the human thyroid, but rather is a synthetic hormone like substance with radically different structures. See the image below and functional properties. And if you look at the structure actually this is just a KQLA structure where all the bonds are approximated as 90 degrees and this is the actual wine bond structure. The structures are identical, the only difference is that one is a sodium salt and the other is the free acid. So as a chemist I read this blog and the thing that comes to mind is like this person is an idiot, I will never go to this website again, you've lost all credibility. This is why organic chemistry is important folks. So back to this example and I'll get to paleo shortly. Here's L-thyroxine, this is T4, the hormone. If I grab the mirror image I make this molecule, I flip that 180 degrees, you can see that they're enantiomers, they're non super opposable. In this structure the wedge in the case of nitrogen is coming at you and this structure is going away from the board. This has cardiac side effects, this is the hormone T4 and this is merely here the sodium salts of the structures, as you can see are identical. So my point here is that very small changes in structure are going to have huge changes in function even if you're merely changing the orientation of the molecules in three-dimensional space and this is important when it comes to talking about other things. Elatic acid again and oleic acid, the only difference between the two is the geometry of the double bond. Elatic acid is going to increase cholesterol as to transfer protein activity, it's going to induce dyslipidemia, it impairs endothelium function, decreases insulin sensitivity, resoleic acid has been shown to improve glycemic tolerance through increased secretion of glucagon like peptide 1 and it reduces blood pressure. Double bond geometry is the only difference. If all you know is the name, this is completely lost on you. You have no clue. Since Dr. Lustig just gave his talk, I thought I'd point this out, the difference between glucose and fructose. Now it turns out that I'm cheating a little bit because in solution what you really have are the two anomers, gluca and beta, as for fructose you've got fructo-pyrinose and what you really need to do to compare them properly is actually draw the linear structure which doesn't exist in high concentrations in solution but nevertheless bear with me. You can see this is an aldehyde, this is an aldose, a hexose and this is glucose. The only difference between glucose and fructose, fructose being a ketose but also a hexose, is that there's a one-two hydride shift, that's not how you would interconvert them but in this case in fructose your double bond is here, in glucose your double bond is here, that is the only difference between the two which is fairly minor but the activity is very different, they're metabolized differently, they can be interconverted but they're metabolized differently, of course depending on the dose. Something else here is adrenaline biosynthesis, you can take this phenethylamine backbone, this is phenylalanine here, hydroxylated, you now have a completely different amino acid with different function just by adding one hydroxyl group, so now a really small change in structure is going to have a big change in its the activity, you add another hydroxy group, you've got aldopa, you decarboxylate that, you've got dopamine, you hydroxylate that, you then have norepinephrine and then you methylate that and you've got adrenaline, so again some pretty minor changes in structure is going to give you some huge changes in the activity and of course chemists have banked on this structure, this basic phenethylamine backbone to make a variety of very interesting molecules, some of the better known are unfortunately probably the street drugs, mescaline and crystal math, so here's how I would approach this question in a chemistry department, I'd start talking about natural products, organic chemists devote an entire field to the isolation, characterization and determination of biological activity of natural products, many of which come from plants, so when it comes to plants you've got cellulose which serve the structural purpose, you've got things like capsaicin which serve a defense purpose and then of the biologically active molecules most people know caffeine, I'm hoping that most of you are well caffeinated this morning and maybe no taxol which is packly taxel isolated from the needles of the U-tree, this is a mitotopic inhibitor that's used in cancer therapy, however plants do synthesize compounds that aren't always defense chemicals, they can be problematic and those compounds are referred to as anti-nutritional factors, there are substances that can reduce absorption and utilization of nutrients, the efficiency of digestion and they may produce undesirable health effects, but what's interesting is that whether or not a substance is an anti-nutrient depends on the species ingesting the substance because it depends on the digestion process, so something that's an anti-nutrient for a rat might not be an anti-nutrient for a human being vice versa, of the known anti- nutrients you've got things like tenons, so proto-anthocyanidins, those are forage legumes such as white clover, chlorogenic acids, that's a digestive enzyme inhibitor that's found in sunflower seeds, you have toxic amino acids like cannabinine and gingolic acids that are found in beans, you have phytates and phytic acid, these bind iron, calcium, magnesium, zinc and copper and prevent you from absorbing them and they also inhibit digestive enzymes, you have cyanogenic glycosides that release HCN and incredibly toxic substance, those are found in cassava, limey beans and flax, then amygdalin, bitter almonds as a source, oxalate is another metal chelator, you have FODMAPs which are fermentable, oligo, dye, mono, saccharides and polyols, those can be found in places like beans for example, raffinosis of galactolygosaccharide, another great word for scrabble, and then you've got xylitol which is found in berries and oats and mushrooms and that's a polyol, some of them that I'm going to be talking about specifically are saponins and glycoalkaloids, prolemines and proteins and a few others, so the reason why I'm beating this whole structure and function thing over your head is because of this, if you look at the anti-nutrient content of say rice and maize which is corn, of course, you're going to see that in both species it's about less than nine migs per mil, except that corn has a lot of phytate, but if you know you cut it off here, you're going to see that they're in the same order of magnitude, if you look at things that athletes would be eating on a quote unquote paleo diet, you've got yams and you've got cassava, and if you look at the anti-nutrient content, it's the same order of magnitude, they're less than nine migs, same for here unless the cassava contains a lot of cyanogenic legicides, but you cut it off here, right, same order of magnitude, so if you're going to tell someone, hey, you should not eat grains and legumes because they contain anti-nutrients, a biologist, a plant biologist is going to look at you and say wow, this guy's a moron, right, this stuff is really important and you're going to lose credibility immediately if you say stuff like that, you make statements like that, so it's not the right way to sell it, you have to you know evaluate these things on a one-on-one basis. Rice, corn, yam and cassava all contain very similar amount of chymotrypsin inhibitors but you know the cowpeas, the legumes they contain quite a bit more, so I will give you that. So let's talk about saponins. Saponins are glycosides of terpenes, which means that you are hooking up sugar molecules onto terpenes and when you delete all the sugar molecules, what you're left with is egg-alicone, so if you hear this term egg-alicone, it just means the molecule without the sugar. In this case we've got gores of pogenin here, the saponins are typically, or I should say the terpenes are typically made by coupling IPP, which is isopentenol diphosphate, with dimethyl allyl diphosphate. That gives you gerilal diphosphate and this thing is going to be cyclized into virus molecules that contain 10 carbon atoms and we call these the monoturpenes. You can then add another IPP unit and make pharnazyl diphosphate, that's going to be cyclized into molecules that are going to contain 15 carbon atoms and we're going to call those the sesquiturpenes. You can also add another IPP unit and make the di-terpenes with C20 or you can couple two of these, make the squaling C30 and that's going to get cyclized into, epoxidized and cyclized into lanosterol and you can modify that to cholesterol and a variety of other terpenes. The reason why I say this is the terpenes are always C10, C15, C20, C30. They are different than glycoalkaloids. Glyco alkaloids are similar in that they also have the glycosides licked on to them but they're not terpenes, they're alkaloids. They contain basic nitrogen atoms. So if you go up in front of an audience, specifically a chemistry audience, and you mix up these two, again you just lost credibility. They're different types of molecules. So let's start with the glycoalkaloids. These things are really good at permeabilizing membranes, may be causing intestinal permeability but mostly they've been tested on erythrocytes with the red blood cells. This paper here is an in vitro paper and to test them on cacotu cells. Cacotu cells are a type of intestinal cancer that have been manipulated to look like in terrocytes which are the cells in our gut. But I don't like this stuff because it's in vitro but at least they use cacotu cells. Their previous papers were done with multilambular vesicles that have a questionable application to real life. And the reason why I don't like the in vitro studies is because they don't answer a variety of questions that I personally ask myself when I'm trying to evaluate the danger of an anti-nutrient. And here are some of the questions. Does processing such as hulling, soaking, germination, fermentation or cooking eliminate degrade or denature the compound or modify its activity? All that's very important. Does digestion degrade the compound or modify its activity? Do in vitro studies employ physiologically relevant concentrations or amounts of anti-nutrients that are on par with what is present in food or diet. And there's the rub. Most of the time they employ really large concentrations that are not physiologically relevant because they're in the business of looking for an effect. So if you're going to take something that you see in vitro and then assume that it implies in vivo, no. Again, you're going to lose all credibility if you do that. Is the amount of compound that makes it to the gut after processing in digestion sufficient to be problematic? It might not be. Does the compound inhibit digestive enzymes in the process? Does the compound cause intestinal damage? Is the compound or its metabolites enter the blood stream? And are they problematic once they do enter the blood stream? Are there any synergistic effects between anti-nutrients? Has the compound been studied as part of food or in isolation? And has the effect of the compound been studied in humans or just in other species? Remember, anti-nutrients are species specific. So here is a paper where they feed potato alkaloids and their egg glycones or sapogenins, if you will. And they find that when you're feeding the sapogenins, and of course they're using ridiculously large quantities here, and they're feeding them pure, not as part of food. There's an increase in liver weight, but they find that that increase in liver weight is reversible, and this was done in rats. So really no direct testing of increased intestinal permeability. What has been done is that alpha chaconin and alpha solanine mixtures have been fed to mice that are models of IBD, intestinal bowel disorders. And in that case, they do see an increase in intestinal permeability, which can be problematic. I can't, I'm sorry, I can't go into the details of why that is and why that can precipitate inflammation or autoimmunity, but Rob touched on that a little bit yesterday. But what they do find is that intestinal permeability was not affected in the controls. So the best you can say is that this is species specific, assuming that you can extrapolate to humans. They also did this again with another type of model of IBD, and this time the mice were fed potato skins and fried canola oil, adelabitim. So the best that you can say, and of course these compounds, these glycoalkaloids of potato, they're concentrated in the skin except for one species, which is called the Snowden, which is only sent for a potato chip manufacturer. So the best that I can say if this, you know, translates to human beings is that, you know, people who have IBD shouldn't be eating fried potato skins every day. Right? I can't say these things are bad for everybody and no one should be eating them. You understand? So be honest about the limitations of the research that you're quoting. If you overstate yourself, core scientists are going to smack you real hard. It's not going to be pleasant. So here's a really good review about the biological actions of suponins. I'm going to talk about the suponins now. Some of which are good, some of which are bad. Remember again, structure and activity. Some of them are anti-carcinogenic. Some of them increase permeability. So here's one of the papers I'm going to start with the human study where they're actually taking an extract of jinsen. So they're concentrating it, right? Because they're extracting it. This is not food and they're feeding it to people in large quantities. So if you're interested in the structure, as I'm sure no one is, but I am, here's the structures of some of the suponins that are found in this root in Jinsen. And what they do find is that the metabolites, that is some of the molecules where the sugars have been cleaned off, actually do make their way into the bloodstream. They do make their way into concentrations of about 0.3 to 5.1 micrograms where they're feeding them a decent amount of this extract. So for an 80 kilogram human, this would be 12 grams of Jinsen extract, right? So remember the dose question. This is not something that would happen in reality. But why are they interested in this? Well, it turns out that these compounds are anti-carcinogenic. They're not necessarily bad. So again, you have to look at the dose. You have to look at the activity. Why are they doing this? And you think that if all suponins were bad, you know, someone comes up and wants to, you want to look like an extremist. You can say all suponins are bad and immediately you're not taken seriously. These things can have good actions. And you think that they would have seen any detrimental effects from the suponin at that concentration, but they did not. So there are a variety of rat studies, but unfortunately some of them don't use suponins that we consume. So this one uses gypsophila. Gypsophila comes from baby's breath. This is the structure of the gypsogenin and of one of the suponins. And this is not something that we eat. But they did find that there was a change in villus morphology that was observed. However, there was also evidence of an increased rate of mucosal cell proliferation. Serum cholesterol levels were significantly lower. And that's because these things interact because they're similar to cholesterol. They interact with a variety of sterols, both in free flow and solution, and in the membranes. And that's how they interact with the membranes. So if you're consuming this as part of food, the effect is going to be different. And that's another question you have to ask yourself. When I'm not giving ridiculously large concentrations of this material, when I'm actually feeding it as part of food, what happens? And here it looks like, you know, not that much. The animals here were protected because there's just an increased cell proliferation. They were repairing themselves pretty well. That can be problematic for digestive enzymes, though. The quinoa suponins were also studied. In this case, there really isn't much that you can get out of here. They're essentially feeding unprocessed bitter quinoa to rats. And what they find is that the rats have an aversion to it. Well, of course, they do. These things are really bitter, so they're not going to eat and they're going to lose some weight. You know, it doesn't taste very good. However, processing quinoa during the manufacture of an infant cereal reduced the concentration of the membrane-branolinic activity of suponins and increased the palatability and nutritional quality of the cereal product to a level similar to that of wheat-based cereal products. I'm still not sure if that's a good thing. And then there's quilaja suponin. Quilaja is the extract of the bark of the quilaja tree. And this thing is interesting. Here are the structures. That's what I think is interesting. QS7 is the most active one, and you also have something called quilA, which is a crude extract and a mixture of all the suponins that are in there. And what they find is that this thing is a great adjuvant. It is great at priming the immune system such that you can give an oral vaccine and make it effective. In this case, it was against rabies. They also reproduced this with measles. Now, you might be thinking, hey, I got you, Matt. I need to ask some good questions. This is not in food, so I don't need to care. Actually, it is in food. Crude suponins are routinely used by the food and beverage industry at concentrations greater than those required for adjuvanticity. And as such, they have a better safety profile than bacterial endotoxins. These things quilaja suponin is an emulsifier and it's added in soda all the time. So if you drink soda, you're actually getting enough to cause this reaction. And it's also added in a variety of other junk foods. So this is actually something that needs to be followed. But the question remains, is there going to be molecular mimicry? Is this going to precipitate an autoimmune disease when you are not ingesting things such as myelin, oligodendrocyte, glycoprotein, which is going to give you an animal model of multiple sclerosis? We don't know. We really don't know what happens in humans yet. So what about things that we find in food? And then we have to go back to some in vitro studies. And here's one in vitro study. Again, this in vitro study uses some gypsophila. It also uses soapwort, which is saponaria fissionalis. These are the structures of those compounds. It uses alphatomatine, which is found in tomatoes. And it also uses soya suponins. However, the only relevant data point here are the soya suponins because they're the only ones that are found in food and decent concentrations. And the reason why alphatomatine is not very relevant is that it's only found in high concentrations in green tomatoes and not red tomatoes. You can see that the numbers are abysmal for red tomatoes and not that many people eat salsa verde on a large quantity every day. And what they do find is that the effect on the transmural potential difference is really small. And they say that it's small because it turns out that this particular suponin, the suponins that are found in soy, don't interact with the cholesterol membranes. So they're not all that problematic. And this is not the only one. It turns out that the suponins that are also in alfalfa don't interact with the cholesterol membranes so they tend to be less problematic. So that's it for soy. If I go back to this biological action of suponins review, I read this very interesting quote. So the mechanism of action of suponins on the intestinal membranes in vivo is not yet clearly understood. Ingested suponins are exposed to many potential ligands in the intestine, such as bile salts, dietary cholesterol, and membrane sterols of the mucosal cells in anti-nutrients or anti-nutrients in food, nutrients or anti-nutrients, sorry, all of which may reduce or enhance their effectiveness. It could enhance it. It also remains to be confirmed whether traces of the compound itself enter the body through the permeabilized membranes, even though all of the evidence until now points to their non-absorption. Absorption of suponin metabolites though has been shown, and we've talked about that. So it still remains to be determined whether or not this is problematic. We don't know. However, if you read this paper, it looks like this time the soy suponins were problematic. And the only difference is, well actually there are two differences. One of them is that they used twice the concentration as the other in vitro paper I just talked about. The other one is that they used a different species. In this case, they're looking at rabbit as opposed to rat. And as I mentioned earlier, these things are species specific. But what I want to talk about here is I want to use this as a segue to talk about lectins. Lectins are also anti-nutrients that are found in grains and legumes, and they're carbohydrate binding proteins that have one non-catalytic domain. And they're often found in storage organs such as seeds and tubers. And they're characterized by the type of sugar that they bind. As you can tell, they're found in a variety of foods. So potatoes, zucchini, lentil sprouts. I didn't know lentils were in the vegetable category. Fruits such as pomegranate, raspberries, blackberries, all that stuff. And of course a variety of grains. You can go into spices such as allspice, mushrooms. Again, another variety of grains. Corn, rice, barley, all of that stuff. Sweet pea, navy beans, apples, watermelon, coffee, cocoa, coconut, you know, they're all over the place. And if you read this paper in Toxicon, anti-nutritional properties of plant lectins, it answers a lot of the questions that I like to ask myself. These things survive the digestion process. They bind to the enterocytes. They get into the bloodstream. They are highly allergenic. It really seems after reading this review that food is out to kill us. And I just, I'm reading this review. I'm saying this simply cannot be the case. Something's going on here. And of course it turns out that not all of these lectins are bad. Only the ones that combine specifically to the enterocytes surfaces are bad because they get endocytosize and exocytosize. And then there's something else. It turns out that some of the worst lectins that are the ones that are found in grains and legumes are deactivated by heat. So wheat germaglutinin is deactivated by heat. And I think the most relevant data point here is this whole meal pasta that was enriched with wheat germaglutinin after cooking just like normal pasta, not detected. Wheat germaglutinin is no longer detected here. The destruction of the lectins and legumes, red kidney beans in this case, has been studied. What's interesting here is that it increases if you undercook them at 80 degrees Celsius. But if you cook them at 100 degrees or 90 degrees Celsius, you completely destroy these things. The kinetics of this process, if you're a nerd like me, has been studied. You can find it here. And the lectins and runner beans have also been shown to be destroyed by heat in the same way. So it's no surprise that you can find papers like this that talk about this one instance in a hospital where they had a healthy eating day and people ate undercooked beans and they all got sick. Well, yes, it turns out that that increases the lectin content, but most people don't eat undercooked beans. They eat cooked beans and you can tell that if you cook your beans properly, the lectin content is virtually nil. So lectins from legumes are not problematic to rats when they're fed at less than 1%. It remains very questionable whether or not these things are problematic in well-prepared food. Here's another lectin, the one that's found in soy. This one is also destroyed by heat. And it turns out that soybean agglutinin is inactivated by pepsin as well. And the goitrogensins, goitrogensins soy, have been shown to be heat sensitive as well. Other heat-label lectins or anti-nutrients, peas, redgram, and lentils all contain measurable amounts of lectin, but when you heat them up and you compare the growth of animals on the raw foods versus the cooked foods, there's very little difference indicating that the anti-nutrients in those foods are probably not all that problematic. So again, huge question as to which one of these are problematic. You shouldn't make broad statements like all lectins are bad. A plant biologist is gonna think you're an idiot and not pay attention to what you have to say. This is not the right way to approach the question. There are some that are really problematic. Here's a study where there's seven patients, two of which were fed raw legumes and then raw peanuts, sorry, and then five that were fed roasted salted peanuts. And you can tell that all but in one case, the lectin actually makes it into the bloodstream from the food. It's not surprising that peanuts are one of the biggest allergy culprits out there. So some of them are obviously problematic even after being cooked, but that's not all of them. What's really interesting too is that you can find papers like this where they're co-administering sugar with the raw, in this case the raw legumes, so there's plenty of lectin and they show that the effects, the detrimental health effects are lessened substantially. Brings me back to the same darn question. What happens when this stuff is administered with food? It's part of food. What happens when you cook it, the concentration is low and you're eating food that's gonna contain some sugars and other things that can interact with it? You don't know. Here's one here talking about the lectins in quinoa and they actually isolate two different fractions. They show that one is active in celiac disease and the other is not, but they run a really cool experiment in that they combine the two and they show that the combination of the two is not active in celiac. Again, what happens when you eat these things as part of food? And then this is a paper I was talking about, Sephan Guinea on Friday, that really is misinterpreted all the time. It turns out that these people are using the fact that lectins can bind to the leptin receptor as proof that it can cause leptin resistance. Well, it turns out that these lectins are used by biochemists all the time because they're really useful for their sugar-binding property. But it doesn't mean that just because of that, it causes leptin resistance. The stuff has to get into your bloodstream and like I said, wheat germ aglutinin gets destroyed by the cooking process. So the only way that you could get about 80 micrograms per mil of this leptin in your bloodstream is if you were to eat large quantities of wheat germ aglutinin on a daily basis and most people don't do that. So this is not realistic to throw out there. It's, oh, these lectins are bad because they cause leptin resistance. That wasn't even the goal of the paper. They were just using this leptin as a tool to study the receptor. So I'm gonna end with a prolemines and then some fermentable saccharides. But prolemines here are a class of compounds that are actually not defense chemicals. They are merely storage proteins. They're nitrogen storage proteins, mostly for seeds. And here you see gluten and the highlighted parts are actually the undigested peptides that can be very problematic. They have various biological activities. There has been some discussion as to whether or not gluten is digested all the time or only partially digested. And I've had this discussion with Elysio Fasano. I showed him this paper and he disagreed with this paper quite a bit. He said he was not able to reproduce these results and he has papers that refute this result. And it turns out that if you look at the fermentation of gluten with lactobacilli and a variety of other fungi, it takes a long time for this protein to be destroyed. So this is a really tough protein. It is very difficult to digest. And the reason why it's bad is because our prolyl-olegopeptidases, dipeptidyl-peptidase 4 and any other enzymes that can cleave next to proline. So these proline proteins are rich in proline. The enzymes, ours aren't very strong. So this thing gets partially digested. It binds to CXCR3. It releases zonulin. Zonulin binds to PAR2 and to EGFR and that increases intestinal permeability by dissolving the tight junction. So we've got the paracellular pathway that's being hijacked here. And then down here, you've got all kinds of autoimmune reactions that are going on that can precipitate inflammation and lead to or contribute to the metabolic syndrome and they can also precipitate autoimmune diseases, but I'm not gonna go into the entire mechanism of that. It turns out that this is not something that people who don't have celiac disease need to worry about because there's very little zonulin release and the tight junctions aren't open for very long when they are open. So again, we are looking at something that is population dependent. The problem is that in this case, you can make the argument that the population, it's populations dependent and the populations are growing. The number of people that can be affected by this is growing substantially. So you've got things, actually you've got things that are gluten sensitivity and you've got things that are celiac disease that's been shown that the mechanisms are slightly different. So had the types of antibodies that you can find. This is another paper that I'm gonna skip. So you have things like neuropathy, for example, that are associated with gluten sensitivity associated, right? Headaches and CNS white matter, schizophrenia. Here's another one, the schizophrenia, both of them showing, right? Individuals with recent onset psychosis and with multi-episode schizophrenia have increased antibodies to glide in, may share some immunologic features of celiac disease, but their immune response to glide in defers from that celiac disease. So there are multiple ways in which this gluten can be problematic. Schizophrenia, again, we have dermatitis hepatiformis, type one diabetes, which one is this? This is IBD. There is a problem with this paper, though. I'm sure Chris Masterjohn is going to pipe up about it. And then there's also associations with rheumatoid arthritis, of course, and a vegan diet. I don't think it's the vegan part, but a diet that's free of this stuff has been shown to have some improvements in arthritis. And I think there's a couple papers there. This is an interesting paper, discusses a lot of the diseases that I've already mentioned, and it says that the strength of the evidence for the use of a gluten-free diet in these non-celiac diseases varies, and future research may better define the benefits. I absolutely agree. Then that brings us to FODMAPs, these fermentable carbohydrates, which I think are being underrated. So in this category, you've got fructose. If your Glute 5 receptor in your gut is not sufficiently up-regulated, it's going to just be fermented by the gut. So fructose can act as a fermentable carbohydrate, so can lactose if you don't have lactase, and that's why people who are lactase intolerant have some problems. You've got fructans, inulins, fructoligosaccharides. Inulin is added to all kinds of products because people think it's good, it's not good. Levons, lactose, polyols, all of these polyols. This stuff is found in all kinds of junk food, like quote-unquote low-carb junk food. Galactoligosaccharides like raffinose, stachios, and verboscos are found in legumes, and that's why legumes make you fart. I'm actually going to skip this paper, but it's just a paper showing that most of the inulin oligofructose that we get is coming from wheat in the diet. The reason why this stuff is bad is because it turns out that it causes the growth of bacteria in the gut, and leads to small intestinal bacterial overgrowth, and it's been shown that the gram-negative bacteria, and to some extent, the gram-positive bacteria too, are really problematic. The gram-negative ones are really problematic because they contain something called lipopolysaccharides in their membranes that makes it through the gut if you have a leaky gut, of course, and it precipitates a variety of immune responses that are really bad. In the case of the gram-positive, you have the peptidoglycans that can also be problematic. This is population-specific to an extent, but it's been shown that hickory, which contains probably one of the highest concentrations of these fermentable carbohydrates, can be problematic even for people who do not have irritable bowel disease, for example. But this has been shown as a study here to be very, very relevant for folks with irritable bowel disease. My one problem with this, actually I have two problems. One, the people who originally came up with the FODMAT theory are not giving enough credit to Elaine Goshall's work, and two, because it's highly inspired from it, two, they're actually using real food, which is good for them, but real food is multivariate. So when you're removing wheat, you're not just removing fructans. You're removing a variety of other anti-nutrients or problematic compounds. So this gets me to the end, some people are probably gonna ask, well, who cares, because we know that when we remove this stuff from the diet, we get benefits, and it's true. There's these trials that are out, there's about three of them that I'm gonna show, and actually Rob showed these yesterday, so I'm not gonna show them again, where you see some really good improvements when you put people on a paleo diet that's free of grains, legumes, and dairy. The problem, of course, is that there is some population dependencies there, and I wanna get back to this food, nutrition, and evolution, where there's, in chapter one, there's this statement, man might be described basically as a hunter-gatherer who has had too little time in which to adapt fully to the new nutritional patterns developed within recent food production technology. So I wanna get back to this statement that people say, we have evolved, and that's a royal we, we have evolved over millions of years, not consuming these foods, hence we are not adapted to them. I think that statement is false, but I'd like to modify it by saying, there has been insufficient time and evolutionary pressure for a complete adaptation to seed consumption to arise in Homo sapiens. That, I think, is a very fair statement, and if you were to say that in the core scientific audience, you would be respected, I'm pretty sure. So why not? And that made me think about the vegetarian argument. There's a lot of vegetarians that will say, you know, we're not meant to eat meat because we don't have claws and sharp teeth. Well, guess what, it's because we evolved with tools. That's why we don't have claws and sharp teeth. Why are we not adapted to all of the anti-nutrients in these things? Because we evolved cooking, folks, and cooking not only increases digestibility by starting the digestive process and starting to decompose on the proteins and carbohydrates, but it also improves it by destroying some of the anti-nutrients. And of course, there's some really great work that's come out of Rangum's work for that. So I'd just like to finish with this bell curve, and I think that this is a better way to present this material and has to do with the statement, is related to the statement that I made before. And this is probably what you're going to find. You're gonna find very intolerant individuals on one end that are gonna be the minority, intolerant individuals here, and most of the population is probably gonna be somewhat intolerant. With very tolerant people, again, being the exception. So when it comes to grains and legumes, they should not be pushed as, you know, the end all be all foods and the most healthy things because it is very likely given, you know, evolutionary pressure and whatnot that a lot of people are still very intolerant to this stuff. So individuals that tolerate grains and legumes, again, should be considered the minority, not the majority. And again, I think that's a statement that would be respected by a core scientific audience. So, you know, of course, you know, if you were a part of the Weston A Price Foundation, or if you're looking at genetics or a variety of things, you can start classifying these things as, you know, more problematic and less problematic. And it turns out that rice and oats have very low concentrations of prolamine proteins and their prolamine proteins resemble those of legumes a whole lot more. And if a scientist were to, at the end of my talk, a core scientist were to say this, I'd be like, I can't argue with you. I just can't, right? Now, when you actually factor in the sustainability of agriculture or the non-sustainability of agricultural culture for grains and maybe even the high like omega-6 content, the low nutrient density, then you can make an argument that we should probably not be consuming grains or that if we are, maybe they should be fermented and they should probably not be a large part of the diet. Again, a much more measured statement that's not going to get you pigeonholed as an extremist. But what about legumes? I don't know. You know, they have different prolamine proteins. In fact, they have globulins in their case because they're part of that cotyledon family. And like I said, the lectins are destroyed by heat. They have to be well-prepared, but their agriculture might be sustainable or often used to enrich the soil with nitrogen. I just don't have an answer there. And if a scientist were to ask me that question, I don't know. Like it's not clear to me. So I hope, and I'm pretty sure this is my last slide, I hope that I have given you information that's going to help you move forward with this. If your movement, and I say your, because I don't consider myself as being part of it, I don't rely on this for a living. I have a day job. If your movement is going to move forward, it will have to be taken seriously by core scientists. And if it is to be taken seriously by core scientists, then you should present it in this term. The reason why this is also useful is because scientists love to be handed projects on a silver platter. It's like, oh, you want more information here? There's information lacking there. Excellent. We're going to start looking there. But if you tell someone, ah, this is solved. All lectins are bad, it doesn't matter. No one's going to pay attention to you. So I hope this has been helpful, and I'll answer any questions. Sorry about that. But Matt will be around, and he's not as unapproachable as he looks. You just got to be not afraid of his bark.