 Thank you so much, Beth. Thank you so much everyone. Can you all hear me? Yes, okay. Today, I'd like to share with you some of my thoughts on oxidative stress and carbohydrate intolerance. In particular, I'd like to share with you some ideas that are out there in the literature, but maybe are not really fresh in people's minds when they think about these things, especially the idea that oxidative stress is an important component of cellular signaling and that it communicates energy overload when cells are overloaded with more energy than they can tolerate and thereby sends a proper signal to decrease energy uptake into the cell and one form that that can take because glucose is an important energy molecule is decreased retention of glucose in the cell which could take the form of decreased glucose uptake or decreased glucose output and that can in turn lead to elevated blood sugar, glucose intolerance and thus carbohydrate intolerance and I'll also share with you some ideas that would suggest that oxidative stress can also be a form of miscommunication sometimes and when I get into that I'll share with you some of the findings briefly of part of my doctoral research But before I get into all of that I would just like to briefly kind of set the framework that we're dealing with by addressing the question of whether humans are adapted to consuming carbohydrate mainly because This is what kind of renders the rest of my talk meaningful in other words If humans are not adapted to consuming carbohydrate at all then it wouldn't be very meaningful to ask the question What causes glucose intolerance because the answer would just be that humans don't tolerate glucose so Clearly it would take me much more than a 40 minute talk to comprehensively address this question and that's not my attention I would just like to offer a couple very brief illustrative examples of why I'm working on these questions from the starting point of assuming that humans should be able to Tolerate consuming carbohydrate in the absence of particular pathologies One of the most interesting cases of a carbohydrate specific adaptation that humans have is the duplications in the gene for salivary amylase and salivary amylase is an enzyme in our saliva that digests starch into sugar and Basically when we first eat a bite of starch Immediately the salivary amylase starts turning some of that starch into sugar. So this as far as we know this enzyme is specific to starch So duplications of that and of that gene for that enzyme are an adaptation that is specific to starch Which is a carbohydrate and is thus this is a carbohydrate specific adaptation on the left we can see the number of copies of this gene in 15 wild caught West African common chimpanzees and Each bar represents a chimpanzee an individual one and the height of the bar Indicates the number of copies they have and as you can see they all have two copies one from their mother and one from the father Indicating that in this sample of chimpanzees there aren't any duplications of the gene for salivary amylase on their right We see a totally different picture in the case of humans and we see this from eight different human populations Across the globe which are broken down into low starch eaters and high starch eaters the high starch eaters are Japanese European Americans and Hadza who consume Starches is a major portion of their carbohydrate and the others are either hunter-gatherers or pastoralists who eat Mainly meat or fish and their carbohydrates come from things like milk fruit and honey things that don't have any starch And what we see this graph is organized differently than the one on the left as we go up across to the right we see increasing numbers of duplications for the salivary amylase gene and The higher the bar is this indicates that the more The higher proportion of people who have that number of copies the red bars are the low starch eaters And or the no-star cheaters actually in some cases and the gray bars are the high-star cheaters and what we can see is that unlike chimpanzees Hardly anyone has only two copies of the salivary amylase gene Indicating that well over 95% of humans that have been sampled have at least one duplication in the salivary amylase gene Not only that but we can basically get two things from this graph one of them is that If we look if we imagine the gray bars as a curve We can see that it's shifted slightly towards the right compared to the red bars such that people who do not from populations that don't each starch have The on average are not on average but the the highest proportion of people in that pop those populations has four or five copies of the gene and For high-starch eaters the highest proportions of people have six seven or eight copies So we can see on the one hand we can take questions at the end on the one hand there seems to be some natural selection promoting More copies of this gene among people who traditionally have eaten starch on the other hand It seems like almost everyone has this start specific adaptation regardless of whether they come from a Population that traditionally each starch these investigators Suggested that these duplications may have begun around 200,000 years ago towards the beginning of modern humans They didn't really have really strong evidence about that So we're not going to put a lot of confidence in that dating But what we can say is that most people alive seem to have these very specific starch Start specific adaptations that separate them from chimpanzees and if we look at salivary amylase among great apes We see a very similar pattern So this graph shows not the gene duplications But the activity of the enzyme in the saliva and we can see that humans shown over on the left have far greater Salivary amylase activity compared to any of the other great apes that have been measured here gorillas or rengotanes bonobos and chimpanzees Now the question arises what does this do what does it matter and why do we care? Why would this adaptation be useful and here is a study that came out in the Journal of Nutrition Just this past year that shed some light on it and what what they did was they gave people They tested salivary amylase activity in 48 people and then they took the highest seven and called them the high amylase group And they took the lowest seven they called them the low amylase group And then they got rid of everyone else so that they could see a nice clear difference between the high and low activity so we can try to see what does this enzyme do and What they what they did was they took these people and they gave them It was sort of like a glucose tolerance test, but it was starch instead So they gave them a solution of 50 grams of starch They drink it over 20 minutes and then they measure their plasma glucose and so someone who? Who is well adapted to eating the starch should not have a very large rise in plasma glucose because they should be The glucose should be entering their blood and they should be using it at the same time And there shouldn't be too much disturbance and blood sugar So what we see over here is that for people with low salivary amylase activity The glucose response is much higher than for people with high salivary salivary amylase activity So it's what this shows is that people with high salivary amylase activity are able to tolerate eating starch without Disturbances in their blood sugar the graph on the right shows a possible explanation of why Here we can separate this is the insulin curve and we can separate between the pre-absorptive Which is in the first ten minutes if you look over here? It's ten minutes before glucose really starts rising so this early rise in insulin is The rise in insulin that occurs once you eat the material But before the sugar actually starts entering your blood and then after that is the post-absorptive phase And so what we can see is it's a small difference But the pre-absorptive insulin release was higher with people who had high salivary amylase activity and the post-absorptive Insulin release tended to be lower and what this seems to indicate is that if you start breaking down Starch in your mouth into some sugar then the sugar alerts your body that there's carbohydrate coming in then your body knows And expects the carbohydrate so it responds to it adequately and this picture seems to indicate Sort of the concept that if you it takes a little spending a little money to make a little money In other words if you can spend a little extra insulin in these first ten minutes Then you can save a lot of insulin over here, right? So we start to get the idea that the salivary amylase gene Duplications that seem to be characteristic of humans and separate them from the great eights is something that adapts them to eating starch and allows them to Respond with that early insulin release that prepares their body to handle that carbohydrate load And that's supported by the graph on the left which shows that The greater number of salivary amylase gene copies that people had the greater their cell salivary amylase activity And in fact gene duplications explained 81% of the variation in amylase activity and then over on the right that That salivary amylase activity in turn Explains about half of the variation in that early insulin response and then finally This other part of the study offers some further support to this idea. So here they This starch on the right the starch graph shows Basically the area under the curve of what we already looked at on the left We see the response to glucose So they did the same thing, but they get them 50 grams of glucose instead of 50 grams of starch And we can see that here the gene duplications don't make any difference at all Everyone has the same response to glucose But people who do not have people who have low amylase activity in their saliva have are let I have less Tolerance to starch than they do to glucose Whereas the opposite seems to be true for people with high salivary amylase activity So what this seems to indicate is that most humans have at least some adaptation to starch that allows basically Humans maintain the tolerance to eating simple sugars from things like fruits, but compared to other great apes humans have an Expanded repertoire of carbohydrate because they can also tolerate starch better That's the general picture that emerges here Now if we look back at this original graph, we can basically get two things out of this one There's a lot of variation in the tolerance to starch right look at all over here from two copies to 15 Some people are going to handle starch a lot better than other people on the other hand What we can also see is that almost everyone has some increased Capacity to handle starch when you compare them to say great apes So if we were to summarize these data We could say that between people who come from Populations that have traditionally eaten starch and people who don't there's a bit of difference But on the other hand when you compare them to chimpanzees who don't have any duplications in the amylase gene One of the things that really stands out is that humans as a whole have these carbohydrates carbohydrates specific adaptations Now there's one other thing that that I'd also like to point out Which is that traditional diets among humans that have good health has varied very widely So here if we just narrow in on the Pacific Islands We can see that among people very similar in a very similar diets based on tubers and fruit and coconut and fish But in different proportions, we can see that carbohydrate ranges from 34% to 50% to 69% Among these different groups and I'm sure many of you know that there are other groups that have even more extremes So for example, some people have estimated that the Inuit hardly consume any carbohydrate and other people have Estimated that the Tuca Senta consume over 90% carbohydrate So there seems to be a lot of variation But this indicates to me that humans can thrive on a wide variety of different diets and those diets It seems should be should be able to have some carbohydrate now I don't think this evidence indicates anything about how much Carbohydrate people need to consume and doesn't really tell us anything definitively about who can tolerate carbohydrate And just how much starch people can eat nothing like that. So I'm not trying to make a quantitative Point here. I'm just trying to make the point that it seems like humans should be able to tolerate eating some carbohydrate But then we run into this problem, which is that many people in our society can't only 58% I mean, that's the majority right but still only 58% of our population Does not have any kind of hyperglycemic disorder on the other hand 42% That's approaching half have some type of hyperglycemia whether it be the 13% who have diabetes or the remainder of that who have Pre-diabetes and pre-diabetes can be either elevated blood glucose or it can be glucose intolerance In other words, maybe your fasting glucose is fine But when you eat sugar your blood sugar goes out of whack and in fact we can say that about 25% of Americans have some kind of disorder where when they eat carbohydrate their blood sugar goes out of whack So the question that I'm going to try to ask here is why if we have carbohydrate specific adaptations And if healthy populations consumed anywhere from almost no carbohydrate to almost all carbohydrate Why is it that in our population where we come from one of these populations with higher salivary amylase activity? For example where we come from you know a tradition of brain eating and so on Why is it in our population that we seem to be so intolerant of carbohydrates now? There are a lot of aspects to this particular question. Why is glucose intolerance so common? There are a lot of ways to approach that and I'm not trying I'm not trying to give the answer here but I'd like to try to give part of the answer and My working framework for understanding part of the answer is that it comes down to energy imbalance In a healthy human energy input should be balanced with the capacity to burn or store energy Now this I am not trying to say that this all comes down to people are eating too much That's not what I'm saying at all, but It could be that people are eating too much that could be part of the problem It could be that people don't have the capacity to burn energy That could be part of the problem and it could be that people don't have the capacity to store energy That could be a problem too and in fact there are some animal experiments that show that if you delete a gene that controls the extracellular structure of the adipose cells so that the adipose cells are able to expand Further than they ordinarily would be then you can give rodents the same purified refined high fat diets that usually make them fat And they still get fat, but they don't have any metabolic dysfunction because you've increased the ability to store energy So it's not about people are eating too much It's about somehow the balance is being broken between what we're eating and our capacity to deal with it No matter how we deal with it whether we store it or burn it or whatever so when this balance goes out of whack then I'm going to maintain that there are communication signals that lead to insulin resistance and that insulin resistance is an adaptation In is it not a healthy adaptation, but sort of the best we can do given this situation adaptation to this Disequal disequilibrium between energy input and energy storage and burning and This insulin resistance has this adaptive function because what it does is it decreases the energy input into the cell in other words Each cell is going to say, you know, I can only handle so much energy if I can't handle that energy I'm going to stop taking it in and then what is what does it do? It sits in the blood instead. So blood glucose goes up blood triglycerides go up all these energy molecules accumulate in the blood And I'm going to explain how oxidative stress may contribute to proper communication and may contribute to miscommunication in these signals In order to understand this communication process, we need to understand how insulin normally functions What is the communication function of insulin in the normal state and what we can see here is that it acts on The liver the adipose and the muscle tissue to regulate their balance of energy incoming and outcoming with the blood The liver is an important organ that takes up glucose from the blood But it also is an important organ that makes glucose from amino acids called gluconeogenesis and sends it out into the blood insulin normally increases glucose uptake and suppresses glucose release from the liver and this leads to a net flow of glucose from the blood into the liver Similarly, it also increases the uptake of glucose into adipose tissue and into muscle and this also Increases the flow of glucose from the blood to these tissues However insulin also does a couple other things it increases the synthesis of fat from carbohydrate in the liver and Increases the output of this fat or triglyceride from the liver into the blood it also increases the uptake of these triglycerides into adipose tissue and suppresses the release of fat from adipose tissue and so all of this tends to make triglycerides flow from the liver into the adipose tissue and Through the blood and from glucose from the blood into all these other tissues now. What happens in insulin resistance? Well, it depends which pathway we're talking about So for example, if you look at the pathways promoting glucose uptake from the blood into the tissues Insulin resistance causes a breakdown in all of those pathways However, if we look at triglycerides, we and the excuse me and this leads to an increase in plasma glucose If we look at triglycerides, the situation is a little different Insulin resistance interferes with the uptake of tri triglycerides into adipose tissue However during insulin resistance as it occurs in humans the pathway that causes triglycerides to come from the liver into the blood is maintained Now what this what this does is it increases triglycerides in the blood because the triglycerides are coming out of the liver And they're not going into adipose tissue But we can also see something really really interesting about this pattern and that is that it's selective Insulin resistance doesn't target every pathway equally So if you notice this is the one pathway where it's promoting energy output from one of these organs And that's the pathway that's maintained the other pathways where insulin promotes energy intake into the organs Those all become resistant so this is part of the reason why I believe that insulin resistance is an adaptation to energy overload because when the liver is overloaded with energy it it moves from From responding differently in triglycerides and glucose to basically trying to push all of that stuff out into the blood Regardless of whether it's triglycerides or glucose so insulin resistance is basically stopping the retention of energy in cells That are already overloaded with energy So how does oxidative stress fit into this well first I have to define oxidative stress for you The old definition is that you have to have a general balance between oxidants and antioxidants and when that balance gets out of whack then you get oxidative damage to proteins nucleic acids and lipids and whatever else you have in the cell this everly everything starts getting destroyed in 2006 Dean Jones from Emory University proposed that we redefine oxidative stress and I'm very sympathetic towards his view and I've basically summarized it here with my own graphical depiction this is that oxidants and antioxidants play an important role in cell signaling and in fact We don't just have a general balance, but we have numerous different compartments that all regulate a certain a certain coordinated set of proteins to regulate their function and These proteins functions are regulated by electron transfer Reactions in other words oxidation reduction reactions or redox reactions and When this gets out of balance then we have disrupted communication that leads to improper protein function and Jones isn't saying that oxidative damage doesn't occur But what he's saying is instead that when we have these imbalances and we lead to an over which leads to an overproduction of oxidants that 90% of these oxidants are not free radicals But there are other things like hydrogen peroxide and about 10% are free radicals that can damage molecules However, about 99% of those free radicals get converted into these other Occidents and only a tenth of a percent of the original oxidants are left over to damage macro molecules like proteins lipids and DNA Whereas 99.9% are there to disrupt Cell signaling that's what he's saying now I would take I would expand this and say that it's not always a disruption of cell signaling But some of this is actually a proper communication of the of cell signaling to communicate that energy overload So what I would say is that we need to have energy intake and energy capacity in some sort of balance and When that balance gets thrown off this sends an imbalance that it could that imbalances communicated into the balance of oxidants and antioxidants and The reaction is for that communication signal to tell the cell to do what it needs to do to decrease energy uptake so it's basically communicating that proper signal and so this is important to understand because Basically if cells are overloaded with energy and they're producing signals that stop the influx of energy That's not really a disruption of signaling. Is it? I mean it basically you're just communicating what needs to be communicated Shown here is an example of how this might work So on the left we see an experiment that isn't very realistic It's a test tube experiment, but it demonstrates a certain principle. So what they did was they fed Rats regular diet a high-fat diet or high-fat diet plus a mitochondrial antioxidant I refer to this as diet induced obesity rather than a high-fat diet because there's lots of other things about these diets That are bad and do so be obesity. So I would look at this as a Symptom of obesity or energy whole-body energy overload what they did was they took the muscles from these rats and then they basically they inhibited the Metabolism of energy and then they put in a lot of energy So they're basically creating a maximal state of energy overload in the cells and what you see is Hydrogen peroxide which is an oxidant gets produced when you put in this energy input and in the rats Who are already suffering from obesity that is high whole-body energy overload the production of hydrogen peroxide is greater Okay, now. This is an unrealistic scenario in that they've you know really manipulated this experimentally But it's but it's we need to do this in order to demonstrate the principle sometimes because we're dealing with really tiny Concentrations of oxidants that are often hard to measure. So this demonstrates an important point What we see on the right is a more realistic set of results from the same animals where they gave the animals oral glucose tolerance test and what we can what we see is that as expected the diet induced obesity Increased plasma glucose response in other words a decreased glucose tolerance and an increase the plasma insulin response in other words it Increased insulin resistance. Okay, but what's interesting here is the mitochondrial antioxidant the same one that normalized hydrogen peroxide response to energy overload Normalized the plasma glucose response and the plasma insulin response So this indicates that hydrogen peroxide and oxidant may be carrying forth that signal that Saying to the same the cell is saying hey, I got too much energy. I'm not going to take in any more So if you restore the oxidative capacity to the mitochondria and you get rid of that Excess production of hydrogen peroxide then all of a sudden you can tolerate the glucose and you are sensitive to insulin So if we go back to this schematic that Dean Jones had published I would modify it a little bit to adapt it to this scenario that I'm this picture that I'm trying to paint So I would say let's talk about energy overload is the main cause not that that's the cause of all oxidative stress But that's what we're talking about here some of the free radical intermediates that are produced during energy overload are Superoxide and the hydroxyl radical some of the non radical oxidants are hydrogen peroxide But in fact what we're dealing with is a two-way street here And so the mitochondrial will produce superoxide most of this will get turned into hydrogen peroxide But it's not just that hydrogen peroxide only affects signaling hydrogen peroxide is really dangerous, right? If you have an infection you pour hydrogen peroxide on it. What does it do to the bacteria? It kills them, right? Yeah So hydrogen peroxide is dangerous and that's in large part because of this two-way street because hydrogen peroxide can then produce the hydroxyl radical Which can be damaged molecules, but most of this hydrogen peroxide is going to Act in cell signaling to decrease energy retention But I wouldn't really call that a disruption of cell signaling because what does that do that fixes this part over here So all of a sudden, you know, we start with energy overload But then we produce the hydrogen peroxide the hydrogen peroxide helps say look, I'm overloaded with energy and the cell reacts to this by Normalizing that process of energy overload of course that has negative consequences because if the cell says hey, I don't want that glucose What happens to glucose in the blood? It goes up, right? So that's so it's a problem But it's it's but you know the alternative is for the cell to say I'll take all the energy I want I'll produce all the hydrogen peroxide I want then the cell finds itself in the same situation that that bacteria was in when you Poured the hydrogen peroxide in the cut Okay, so it's so insulin resistance isn't a good thing But there are all our alternatives which are worse which are for the cells to have no regulation of how much energy they take in and That can lead to a lot worse damage than insulin resistance itself leads to So there are a couple questions that are left and that those questions are How does oxidative stress communicate energy overload and this is you know there's a lot of work left to do but I'll show some preliminary thoughts on this and then can it ever be a form of Miscommunication and I'll make the case that sometimes it's sometimes oxidative stress is a form of miscommunication And there are things that we need to do to help regulate everything properly beyond simply fixing that energy balance So one plausible hypothesis of how this might play out This isn't definitive, but this is a general schematic hydrogen peroxide and other oxidants may play a number of roles in cell signaling One thing that they might do is oxidize and deplete glutathione, which is abbreviated throughout these slides as GSH Glutathione is an antioxidant and detoxifier and regulator that we make from proteins So when we eat protein we make glutathione ourselves and it has all kinds of functions including as an antioxidant But also as a regulator of protein function One of the ways glutathione regulates proteins is by directly binding to them so that in conditions of oxidative stress Glutathione will bind to the protein and change its function and that will in turn allow the process of communication to go on But another is that glutathione depletion will lead to the accumulation of methylgloxal And I'm going to share a little bit about this pathway with you First of all because there's a little bit more known about it and second of all because I just did my dissertation on it and And it saved me a lot of time to be able to prepare this talk if I could include some of that information here So hopefully you'll get something out of it too Okay, so methylgloxal, which is abbreviated MGO through these slides is shown in the upper left and it reacts with amino acids to form advanced Glacation and products which are abbreviated here as ages a lot of people blame Advanced Glacation and products on glucose and the misnomer Glacation really really facilitates this unfortunately But in fact most ages in the human body are not produced by glucose directly They're produced by methylgloxal and a couple other similar similar molecules And here is the reaction of methylgloxal with Arginine and amino acid that's found in proteins to produce the most abundant Advanced Glacation and product found in human plasmid in an amyl tissue and one of the things you can see here is that? The positive charge that's usually present on Arginine is gone Once the advanced Glacation and product is formed and if you change the charge of the amino acids in a protein Then all of a sudden you're going to change its shape and if you alter a protein shape. What do you think you alter? It's function right it's just like if you get in the house with your key And then you change the shape of the key that key is not going to work anymore Right, but maybe you'll get into someone else's house, right? So it's not that you necessarily obliterate the function, but you change it right now When we alter protein structure and then alter protein function Most research on this focuses on chronic disease and oftentimes it just skips right from here to here totally Oblivious to the fact that when we alter protein structure and function that allows a system of regulation and Most people don't talk about advanced Glacation and products is regulatory molecules But I think that in fact advanced Glacation and products are regulatory molecules and are involved in communication And here's a little scenario of how that might work So methylglyoxyl can primarily come from two sources one is from glucose Not directly, but through the process of glycolysis glycolysis Lysis is like to cut glycolysis is basically splitting glucose in half And that's the first step in burning it for energy and when we do that we form some methylglyoxyl in that process The other place we get methylglyoxyl is from acetone. Anyone know where acetone comes from? Shown right up here fatty acid metabolism right acetone is produced during ketogenesis So we break down fatty acids for energy We produce some acetone and acetone gets converted into methylglyoxyl now There's two things that happen here. The first is when we produce methylglyoxyl from glycolysis methylglyoxyl inhibits glycolysis Now that's I don't think that's an accident. I think that's a system of negative feedback So that helps keep glycolysis in check if you're breaking down too much glucose You get more methylglyoxyl methylglyoxyl comes back and stops that process just keeps it in check right now The other thing that we see here is fatty acids Ordinarily, you know if you read any textbook on biochemistry. They say you cannot make glucose out of fatty acids And what that means is when you break down a fatty acid into acetyl CoA and you go into the TCA cycle Theoretically you should be able to get carbons that go through this pathway over to make glucose But what happens is every time you bring in two carbons into the TCA cycle two carbons leave as carbon dioxide So you never get a net flow of carbons from fatty acids to glucose However, if you convert fatty acids into acetone into methylglyoxyl You can actually turn methylglyoxyl into glucose So when we shift into fatty acid metabolism, we do two things with methylglyoxyl one We are we're not getting enough glucose because we're burning fatty acids instead So we actually make some glucose from the methylglyoxyl that we make from the fatty acids But then it also comes and inhibits the breakdown of glucose. So we spare some glucose So this indicates that methylglyoxyl plays some legitimate communication functions and some legitimate roles in metabolism Okay, however There also is good evidence that methylglyoxyl plays a role in disease Here we see that methylglyoxyl concentrations are elevated 3.6 fold in people with type 2 diabetes Now that indicates that either diabetes increases methylglyoxyl levels or methylglyoxyl levels Increase the risk of diabetes and we don't really know which one and the data is not there to really tell us But it might be a little bit of both for example if we look at the metabolic pathways regulating methylglyoxyl accumulation we can see that insulin should be one of the primary Primary protectors against the accumulation of methylglyoxyl Just to briefly go through it Methylglyoxyl can be formed from acetone as we said before through this enzyme CYP2E1 It can be produced during glycolysis from these intermediates triosphosphates and when it's detoxified It's detoxified using that master antioxidant of the cell glutathione abbreviated GSH So where does insulin come in? Well insulin decreases the production of acetone Insulin decreases the activity of the enzymes that convert acetone to methylglyoxyl Insulin increases the activity of glycolytic enzymes that clear these intermediates that would otherwise Accumulate and become methylglyoxyl Insulin increases the production of glutathione Insulin increases the production of the first enzyme involved in the detoxification of methylglyoxyl all these indicate that insulin should be the primary protector against the accumulation of methylglyoxyl and the formation of advanced glycation end products and since Diabetes is a deficiency of insulin whether it's a deficiency of insulin itself in type 1 diabetes or Deficiency of insulin signaling is in type 2 diabetes then that could be one reason why methylglyoxyls is increased in diabetes However, there's also good evidence that methylglyoxyl may cause diabetes so here we injected or a group injected methylglyoxyl into rats and It increased the plasma glucose response to a glucose load that can be seen most clearly over here So it it increased glucose intolerance and here a scavenger of methylglyoxyl attenuated this effect Here we see that infusion of rats with methylglyoxyl concentrations over a month Increases the apoptosis of pancreatic beta cells in other words half of the beta cells died in the pancreas which is a hallmark of severe type 2 diabetes and This is seen with the brown stain on the right indicating dying cells And this was consistent with other features of type 2 diabetes like insulin depletion in the pancreas and other things like that So part of my dissertation project was to study whether glutathione being important for methylglyoxyl detoxification Actually means that we can change methylglyoxyl concentrations by changing the levels of glutathione If this is the case then this would indicate that methylglyoxyl could carry out signaling roles of glutathione depletion previous research has shown that toxic chemicals like hydrogen peroxide or other toxic chemicals that deplete glutathione all increase methylglyoxyl concentrations in cells But these were also toxic to the cells and likewise the administration of BSO, which is an inhibitor of glutathione synthesis increases methylglyoxyl concentrations in rats But this was given in the drinking water for a month Which proved also toxic to the animals and that also caused their pancreatic beta cells to start dying and so on So and this shouldn't be too surprising because when you administer this in the drinking water You get an increase in oxidative damage So what I wanted to look at was is there a way that you can deplete glutathione and get this This increase in methylglyoxyl that could communicate a signaling role of oxidative stress without having this damage and toxicity So previous research had demonstrated that if you just do one injection of BSO instead of giving it in the drinking water You get depletion of glutathione shown on the left, but no change in oxidative damage shown on the right and So I took 48 rats and gave them either an injection of this BSO or control just once And I showed that you could decrease glutathione and increase methylglyoxyl But when we measured lipid peroxidation, it wasn't decreased So I would conclude from this that BSO Depleted glutathione in my experiment by in the absence of cellular energy overload And this could be seen as cellular miscommunication right because we said before that when you get obese and You're you have energy overload then you start producing these oxidants that might deplete glutathione Lead to increase methylglyoxyl and lead to insulin resistance, but here these animals weren't fat I just gave them a pharmacological drug that depleted glutathione and that causes the same effect that downstream increase in methylglyoxyl Which could cause insulin resistance and glucose intolerance But the this Pharmalogical inhibition of glutathione Depletes glutathione to similar levels that you could see from other things like if you have a low-protein diet So for example in teacol and Campbell's experiments where he fed rats low-protein diets They had severely depleted glutathione Just like we would see with this farm a lot pharmacological inhibitor And also when rats undergo extended fasting for two days or more they get similar depletions And that's because you are deficient in the amino acids that you need to make the glutathione So if you don't have what you need to regulate the system then oxidative stress can be a form of miscommunication and Deficiencies and a lot of other nutrients Inflammation toxins all of these other things could theoretically cause miscommunication of the oxidative stress signal so Regulating glutathione is very complex. It requires getting adequate protein undenatured whey proteins are very important for Sistine in some cases Glycine from bone broths and other things be vitamins proper endocrine function fruits and vegetables to supply polyphenols and Brought support for a lot of the other antioxidant defenses that can interact with glutathione And so what I would conclude from this and I have a maybe 10 seconds left is that? We should be able to consume carbohydrate and a lot of us can't I think part of this is because we have energy Overload, but there might be a lot of other aspects of to this that need a lot of other research So for example there might be a lot of other things like inflammation and toxins and nutrient deficiencies That lead to that oxidative stress signal without us being fat and that can be a major problem, too and so basically what we need is a lot more research to try to define how We can regulate that proper communication, but I think a lot of what it comes down to is that traditional diet that's dense in nutrients and Gives robust thyroid and insulin another endocrine function. So that's all I have to say and thank you very much Is that okay? She says it's okay if we take questions all the panelists come up But I should warn people that I'm gonna stay for the panel. So then any other Questions will be have to deflected further Chris Hi, so I had a quick question So are you suggesting based on the amylase data that there may be four possible categories for Humans adapting so one group would be more tolerant to starches one group would be more tolerant Maybe to fruit one group would be less tolerant to both of them and one group would be more tolerant to both of them What if you're just asking about the amylase adaptation? I think that indicates that compared to great apes Great, I'm saying within the bell curve of the human within the bell curve I think that indicates that some people can tolerate starch good well And some people can't tolerate tart tolerate starch as well But the amylase doesn't seem to have any implications for toleration of fruit Okay, however, there could be other reasons that people can't tolerate fruit Like for example, maybe they have oxidative stress or you can have digestive problems lots of other things too The other comment is that I've seen a lot of antioxidant research and I haven't really seen any research Suggesting yet, and maybe this will be proven wrong that just giving overweight people antioxidants is going to change Their insulin response or their blood glucose response, and I know I'm maybe oversimplifying But at the same time I do agree with your glue a tie-on argument. I think that's an excellent point Yeah, well part of the problem is that Most of the antioxidant supplementation trials are based on the old paradigm of if you just increase the amount of Antioxidants relative to the amount of oxidants then you solve oxidative stress But I think what this indicates is that it's way more complicated than that. Thank you Okay, we can do one more question So in the beginning I wasn't totally clear as to whether or not the data you had was it genetic Differences in the amylase or was it measured differences based on the function of the amylase? Oh both so the first slide was the Variation in duplications of the gene and the second slide compared the actual activity in the saliva of humans other great apes So they correlated so I wonder oh, they do correlate very strongly. Yeah, so I wonder if this is not one of these epigenetic adaptations that somebody asked about in the beginning you know how does epigenetics fit into these adaptations in the first One of the first lectures we had here that the more that we use enzymes the more of the DNA gets exposed And the more it could potentially get duplicated and I think that's genetic rather than epigenetic But I think you're right that if you have I I don't think that Mutations would be random with respect to use of a gene I think if you have the DNA opened up there more vulnerable mutations, right? So this could be a beautiful example of somebody's answering somebody's question in the audience earlier So thank you for yeah. Thank you very much Okay, thank you everyone