A Microbiome-Based Weight Loss Solution

Hello everyone, it's Satchin Patel and welcome to our presentation. Today we have a special guest, Kiran Krishnan of Microbiome Labs. We're going to be discussing the connection and correlation between the microbiome and weight. This is a very relevant topic right now with everything that's going on in the world, but it's relevant because weight is one of the top concerns that many people have when it comes to their health. And certainly people who are coming into our functional medicine practices or coaching practices, health practices, are looking for some sort of solution for weight loss. And this year in particular, there has been a little spike or maybe a big spike in weight gain for people for a variety of different reasons. People might be more sedentary, they might be eating emotionally a little bit more than before and there's also, you know, a lot more stress going on in people's lives and that can lead to increased weight gain. In our practice we see a lot of weight loss clients and you know, part of their journey is to reduce their inflammation, improve their microbiome health and a lot of other factors when it comes to their health. So today this topic is going to be very of high interest to me myself. I'm looking forward to learning a lot from Quran as I always do. And just before we even started recording, he was dropping some gems. And so I don't leave you all out of that conversation. We're going to kind back to it a little bit later. We were talking about breathing, we were talking about mouth breathing versus nose breathing, how that affects weight, how that affects your overall health. And that's something that is very interesting and relevant to me right now. I'm going down that breathing rabbit hole. So without further ado, welcome. Karan.

Hi, Sachin. Great to be here with you. Thank you so much for having me. By the way. It's a pleasure.

Absolutely. It's always my pleasure because you add so much value into my life personally and it's always great to share your powerful message with the clinicians that we mentor and the clinicians that are in our ecosystem because they get to connect with great cutting edge scientific information and the products that you guys put out there are second to none. So I'm thrilled about this conversation and dialogue that we're going to have and I know that you've got a PowerPoint for us. So I'm going to pass on the host to you so that you can go ahead and share your screen whenever you're ready. But before you do that, as people are getting settled in, as we're letting the technology do its thing. What are you most grateful for right now?

Right now, I'm most grateful for not having been impacted by this pandemic that's going on the same way it has impacted millions of people. I'm extremely grateful that I haven't lost anyone close to me or even had anyone really getting significantly sick. And also, of course, not having been economically impacted like a lot of people have.

It'S sad to flick through news articles day in and day out and kind of see the impact that it's having on people. But it also allows me the opportunity to take stock of the advantages I have because of that. Number one, in my own health, how can I make things improve things to be even healthier? How can I eliminate certain bad habits with this time that I have to spend on myself where I'm not traveling around like a crazy person? And then also, what can I do and my family do to impact our community in a positive way? We live in a very nice mixed community. Mixed, racially mixed, economically.

There are people in all spectrums of the economic.

Spectrum, I guess, in our community. And so we see a lot of our neighbors being impacted by it. So we're really kind of digging into opportunities in the local community to help people that are being affected. So grateful that I'm not and also grateful that there's this opportunity now for me to learn about my community, learn about ways to help, and then, of course, improve things within myself as well.

Awesome. You're such a go giver. And, you know, I'm excited for what you're going to share tonight, so thank you. Let's get right into it, my man.

Yeah, let's do it. Let me jump in here. Sharing.

Okay. And then. Hi, everybody. Everyone that's either watching this as a recording, certainly here live as well. I'm going to go ahead and turn off my camera because I tend to make really strange faces when I talk, so I don't want to be distracted by my own face. I'll do that so I can focus on the words itself. But it's a really great honor to be able to talk about this because as you guys know, metabolic health and weight issues are a significant problem not only in our country, but globally. That has been first further highlighted by this pandemic. Obviously, people who are obese have four times, five times greater risk of succumbing to this pandemic, have a much worse go of this particular virus. And that's because their immune systems are compromised in a significant way. Our immune Systems are perfectly well capable of dealing with something like this. But when we compromise it because of metabolic dysfunction.

We harm ourselves in so many different ways. I also know psychologically for a lot of people to make all kinds of improvements in their lives, it starts with weight and their own body composition and how they feel about themselves. So being able to get that under control really will be the stepping stone for them to make all kinds of other changes in their lives. So we're always looking for other interesting insights on some of the advantages we can take advantage of through reprogramming the gut through the microbiome and how our body responds to food. Call this metabolic reprogramming. Looking at a microbiome based weight loss, and again, this is something you would tack onto everything else that you're already doing. This is not necessarily a replacement for all the other things you do that has had success in the past. This is really a way to get one step deeper into reprogramming their body to respond differently to food. A lot of this comes from this idea that there are certain people that are naturally lean for most of their lives.

We always said that that person had a high metabolism. Well, it's not really true that they have a high metabolism is that their body responds differently to the presence of food and their diets, although may not be perfect or right in any way, they still may not gain weight. There's enough documentation about this to understand that the microbiome really plays a role. So the first thing that's important to note as it comes to weight is obesity, certainly in the United States and North America in general, continues to be a massive problem. The rise.

In obesity prevalence can be realized over the last 20 years. The last two decades, there's been a continuous upward trend in the number of people that would be considered to be obese or overweight. Even within the US we also know that a lot of people claim to be dieting. Somewhere around 93 million people a year, close to 100 million people a year in the latest couple years claim to be dieting. When over 170,000 3 million people within the US adults within the US 15 or older really are actually overweight or obese. So 2/3 of the population is either overweight or obese. You guys also know that a lot is being spent globally on weight loss. It's over $700 billion market as of 2014. The latest numbers are this is reaching a trillion dollar market for weight loss. And this includes things like foods and beverages, drugs, supplements, devices, services and so on. And despite all of that spending, we're really not making any dent in the percentage of people that are obese and continue to be obese over time. Of course, one of the scariest things is the rate of childhood obesity that's growing as well. And again, this pie just shows you what the US market tends to spend on obesity each year. $65 billion. This was a few years ago, but this is a massive, massive market and people are really concerned and really investing in trying to achieve healthy normal metabolic response and they're not getting it. So at the end of the day, to me, if we're doing so much research in obesity, if we're spending so much money on solutions in obesity globally and we're not achieving success, if the, the rate of the epidemic of obesity continues to increase, there really must be a significant missing link. And one of those missing links could be the role of the microbiome in weight management. Because at the moment, when you look at most weight management solutions, none of them address the issues and the dysfunctions within the microbiome that lead to weight gain and metabolic dysfunction. So the role of the microbiome really cannot be overstated in weight management. That's really what I'm going to cover for you today. Now, a lot of this information came about from studying discordant twins. These are identical twins who have exactly the same genetics, but one twin is obese and the other is lean. Some of the studies that started basically took genetically identical germ free mice. So they don't really have a microbiome of their own. Then they would take microbiome samples from the lean twin and the obese twin implanted into half of the mice. And, and then they fed both mice the same exact diet. And they found over and over again that the mouse that had the microbiome implanted from the obese twin always gained weight, despite eating the exact same amount as the mouse that got the lean twins microbiome. And so this isn't a genetic element. This is purely nature. This is purely nurture, not nature. And that's a nurturing of the microbiome from the beginning. What happens to the microbiome of this individual and in which direction it moves in can determine in large part your body's response to food. Now this type of experiment has been repeated numerous times in many different iterations. They would even do things then like taking the lean twins microbiome, putting this mouse through a course of antibiotics to kind of kill down the bacteria, implanting the lean twins microbiome in the obese mouse and then vice versa, the obese twins microbiome into the lean mouse again, still feeding them the same thing. And then you see the reverse. You see the slim mouse now starting to gain weight and the obese mouse now starting to lose weight. So clearly the microbiome plays a significant role. And the composition of the microbiome is really critical when it comes to obesity. The composition is of course, driven by this issue of dysbiosis. Dysbiosis within the microbiome is really an imbalance, as you guys well know. And ultimately that imbalance does alter intestinal permeability. So intestinal permeability is still a major culprit here. And of course I'll show you some of that data as well. And an increase or decrease in specific bacterial groups are really critical in generating this type of dysbiosis. And dysbiosis is of course associated with a number of conditions that you guys always deal with, things like inflammatory bowel disease, C. Diff infection, autoimmune disease, but of course obesity as well. So the pathophysiology of obesity is not a whole lot different from the pathophysiology of other chronic illnesses like autoimmune disorders, inflammatory bowel disease, and so on. When it comes to the microbiome, there's a lot of uniformity in that. One of the big initial observations as it relates to obesity in the microbiome is this whole firmecutes bacteroides ratio. It's reported to be associated with obesity if an individual over the course of time, based on lifestyle factors and dietary factors, if they bump up their Firmecides phylum by 2, 3, 4% and the resulting reduction in bacteroidetes, you will see a metabolic change in that individual and body composition change as well. Of course, ultimately the diet alters the microbial community in the most major way. And so dysbiosis that occurs secondary to a western diet, which tends to be this higher protein, higher sugar, higher fat diet, because this diet increases the firmekides in the gut. That's one of the interpretations as to why people who are obese tend to see a higher level of firmecutes than people who are lean. The idea was there is this western diet, the types of things that you eat within the typical western diet, the fast food diet, the processed food diet, tends to favor the growth of firmechetes, type of bacteria. And because of that, you tend to find people who are obese will have higher levels of firmekades than people who are lean. So that's the initial understanding behind the impact of the microbiome and obesity. Now we can get a little bit deeper because there are more specific players. Because remember, Firmekides and Bacteroidetes are just the phylum that's the largest groups of bacteria. They're hundreds of species within the Firmechides and Bacteroidetes phylum. So it doesn't tell you a whole lot by just saying Firmecides and Bacteroides. So we do want to get a little bit more specific into what is impacting the metabolic health through the micro.

So the first thing is understanding that they are critical keystone species, especially this one, Achromancer mucinophila, that plays a, that plays a critical role in the microbiome's influence on metabolic health and metabolic disease in general. Akkermansia, as any of you have heard me talk, is referred to as a keystone strain. It's a really important strain and it is inversely associated with obesity, diabetes, cardiometabolic disease and low grade inflammation. Akkermansia is a mucin degrading commensal anaerobe. So it lives in the large bowel. It's a strict anaerobe, meaning it cannot live outside of the body, it cannot be grown in factories, it's very difficult to use it as a probiotic. In fact, there's a couple of companies that have been working on it now for a little over 10 years and still have not had success in actually isolating this as a probiotic. It's designed specifically to live in the gut. And Akkermansia mucinophilia is the only species within this genus of Akkermansia. It's a single bacteria in your microbiota. In healthy individuals, especially lean individuals, it can make up up to 5% of the total microbiota. So it's a really important big player. It's demonstrated to reverse high fat diet, induced fat mass gain. A lot of this was of course done in animal studies and adipose tissue inflammation and of course insulin resistance, which is really important. But its effect requires live viable Akkermansia studies have done where they've isolated Akkermansia from the gut microbiota. They've heat killed it and then they've implanted it into humans through probiotic delivery and they don't see any effect. So you need live, functioning Akkermansia, which is incredibly important. I'll talk a little bit more about what Akkermansia does later on. Other species that you're familiar with that also play a Role within metabolic health within the microbiome is Bifidobacterium. Bifidobacterium's presence in the large bowel is inversely correlated with body weight in both humans and animals. And in fact, Bifidobacterium animalis is one of the important ones within the family, within the genus of Bifidobacterium itself. Again, this typical metabolic mouse study, where they feed the mouse a high fat diet, sometimes a high sugar diet, and they find that viable Bifidobacteria administration was shown to reduce liver triglycerides, total cholesterol, total lipid deposition and body weight gain. The important thing here is that the administration of these organisms, Bifidobacteria animalis in this case, or Akkermansia, not only are we looking at fat mass gain, but you see all of the other metabolic benefits as well. Things like reducing triglycerides, total cholesterol, lipid depositions, all of these things are part of the risk factors of metabolic disease and being obese. So seeing those changes are also absolutely critical in understanding whether or not probiotics and certain strains of bacteria have an impact on overall metabolic health and in fact, more specifically, metabolic disease. Another way that Bifidobacteria helps is it's efficient producer of CLA, conjugated linoleic acid. Most of you are familiar with CLAs. They've got a number of studies on weight and metabolic health and fat burning. So the production, the endogenous production of clas in the gut plays an important role in mobilizing fat for metabolic activity. And the CLA production by Bifidobacteria plays a significant role in that. Bifidobacteria also increases the production of short chain fatty acids and a receptor called GLP1. I'll talk about what GLP1 does in a second. But those are critical components. Short chain fatty acids and These receptors, called GLP1, both play a really important role in metabolic response to food. Another group of bacteria, another genus that you're familiar with, is Lactobacilli. Certain Lactobacilli within the gut has also been shown to reduce fat mass in mice. It also produces endogenous cla. And CLA has of course been shown to induce fat burn and fat loss in animals and humans. Humans tend to have low levels of CLA production and are dependent on dietary sources. Now, commensal Lactobacilli provides substantial amounts of CLA from metabolism of linoleic acid. So feeding the body the right substrates will provide the conversion of Linoleic acids from the diet into conjugated linoleic acids. That's one of the main ways that we end up getting it. When you take clas as a dietary supplement, most of it gets absorbed in the small bowel, which in itself has benefit. But most of the large intestine CLA must be generated through colonic microbiota. So viable lactobacilli is a major producer of colonic cla. And this reduces inflammation, improves permeability, all of the things that are associated with obesity in terms of pathophysiological risk factors, right? So the inflammation, the permeability, all of those can be helped by cla. And of course increasing the intake of linoleic acids are going to be an important step in increasing endogenous CLA production. Now, postbiotics are of course critical to metabolic balance within the body and improving metabolic health. Postbiotics are of course compounds, small molecules, other compounds that are made by the microbiome and itself. And of course the most well studied postbiotics in this case are short chain fatty acids, things like butyrate, acetate and propionate. These are the byproduct of gut microbial fermentation. They play numerous roles, like strengthening the intestinal barrier, enhancing the colon health. Of course, colonocytes use butyrate as their main fuel. Bacteria in the gut make use of indigestible polysaccharides that are coming in, of course, your fiber resistant starches and then they, they metabolize them into short chain fatty acids.

Next thing, slides aren't transitioning. Okay. The major end products are of course acetate, propionate, butyrate. Typically within the first few days, the Production is around 60, 20, 20, and that switches because acetate gets produced at a much higher rate in the beginning, the moment you digest food. When I say in the beginning, I mean when the food comes into the system, you get a higher spike of acetate, but then a lot of the acetate and then eventually the propionate also can get converted to butyrate. What's not being absorbed and utilized by the body is actually going to get converted eventually to butyrate. So you tend to see a bit of a butyrate spike on day three or day four. And they were also the short chain fatty acids also lower the intestinal ph, which can then alter the colonic microbiota. That's one way that the short chain fatty acids play an important role in improving diversity and providing balance within the colonic microbiota. They also increase mucosal Resistance against bacteria that tend to be pathogenic to the body. The goblet cells in the intestinal epithelium that produce the mucin, and of course, the mucin is an extremely important barrier struct. Those goblet cells require butyrate in order to produce mucin. Butyrate as part of the short chain fatty acids are absolutely critical to mucosal health and mucosal structure, which then adds the effect of physical resistance against pathogenic bacteria translocating deeper into the mucosa and onto the intestinal epithelium, where they can cause infection. Increased production of butyrate and acetate are also associated with a leaner body for a couple other reasons. One is short chain fatty acids act as signaling molecules. They interact with things called G protein coupled receptors, in particular GPR41 and GPR43. These G protein coupled receptors are expressed on adipocytes and on your intestinal epithelium. And the activation of GPR41 by short chain fatty acids stimulates a release of leptin, which then is important in reducing the appetite. Right? So if you think about when food enters into the system, as it moves from the stomach to the small intestine, then from the small intestine, it enters the large intestine. What should start happening right away is the production of short chain fatty acids. That production of short chain fatty acids will actually trigger the release of leptin by activating GPR41, both on adipocytes and the intestinal epithelium. The release of leptin creates satiety, and it also counteracts the increase in function of ghrelin, the hunger hormone, so it makes you eat less over time. And then leptin also increases metabolic activity. Oedipocytes, for example, when they sense the presence of short chain fatty acids, they release something called adiponectin. And the adiponectin is involved in the upregulation of something called ampk, which is an enzyme that monitors cellular energy, and it stimulates fatty acid oxidation in all of the peripheral tissues, right? So your fat cells have a way of triggering all other cells in the body to burn fat for fuel. So, after a meal, not only are the short chain fatty acids signaling to your brain to stop producing hunger hormone and instead release leptin, the satiety hormone, which kicks in all kinds of metabolic activities. Your adipocytes, when they bind short chain fatty acids, are releasing ampk, which then stimulates all of the peripheral cells in your body, peripheral to the gut to start burning Fat for fuel. So that's one of the ways in which you turn on beta oxidation. That's the cellular process of burning fat for fuel. In obesity, low levels of short chain fatty acids tend to be produced. So you get less adiponectin, which is generated, which means less stimulation of ampk, which means decrease in fatty acid oxidation. And as a result, because you're not burning the fatty acids that are coming in through the diet or being generated from digestion, you end up having an increase in free fatty acids into the liver. Part of the free fatty acid influx into the liver leads to issues like non alcoholic fatty liver disease, which is also then associated with obesity. Now, this creates a plethora of issues because as your liver becomes more compromised, you of course reduce your bile acid pool. And if you're not releasing bile adequately into the small intestine during digestion, you can absorb your fatty acids as well. You can't absorb fat soluble vitamins. You in fact allow for more dysbiosis to occur in the small intestine. Because one of the important jobs of bile is to act as an antimicrobial to prevent the overgrowth of bacteria in the small intestine during the digestive process. So imagine now you've got this free fatty acid flow into the liver. The liver is getting fatty depositions. It's compromising the function of the liver. The liver is now producing less bile. That reduced bile is attenuating your ability to absorb fat soluble vitamins. It's attenuating your ability to absorb fat soluble nutrients. It's also attenuating your ability to remove fat soluble toxins out of the system. That's another important function of bile. It basically conjugates fat soluble toxins, takes it to the liver for removal from the body, and then on top of that, you're losing a really important component of the antimicrobial effect of bile. And then there's a second antimicrobial effect of bile, where bile actually turns on something called the FXR nuclear receptor in the ileum of the small intestine. The turning on of the FXR nuclear receptor stimulates the intestinal epithelium in the small intestine to release antimicrobials into the lumen as well. These are the two really important functions in preventing an overgrowth of bacteria in the small intestine. So now that your metabolic response is compromised, your liver is compromised, your bile secretion is compromised, you've got all kinds of absorption issue, you've got an issue with accumulating fat soluble toxins, not being able to remove them. And at the same time, you're starting to get overgrowth of microbes in your small intestine. You're getting to the early stages of sibo, for example. Now, one of the things that tends to happen when you get this change, studies have shown in people who have non alcoholic fatty liver disease, and thereby because of that, this small intestinal overgrowth, they tend to get a taxa shift in the types of bacteria that are overgrowing in the small bowel. They tend to see higher amounts of gram negative bacteria growing in the small bowel. The small bowel normally is higher in gram positive bacteria. And a lot of these gram positive bacteria are bile tolerant. That's why they're allowed to survive within the small intestine to a certain degree. Now if you start reducing the amount of bile, you start getting more gram negative bacteria growing. And what do gram negative bacteria have? They have lps, lipopolysaccharide or that endotoxin. Right. So now they're releasing more endotoxin into the small intestine. The release of more endotoxin into the small intestine makes the small intestine more permeable because it recruits inflammatory response into the small intestine. That permeability in the small intestine then causes a translocation of more lps, which both goes into the liver, further harms the liver, and then enters circulation and causes all kinds of problems that lead to further metabolic distress. So this is a big cyclical problem. And all of these dysfunctions can lead to other conditions like cell SIBO and autoimmune disease and inflammatory bowel disease and so on. All of it can be connected to this type of dysfunction just from the reduction in the production of short chain fatty acids. Right. So that whole long story I told you, which hopefully made a little bit of sense, all stems from a reduced amount of short chain fatty acid production. So now this person is dealing with SIBO and leaky gut and increased risk for autoimmune disease and all kinds of other stuff on top of not burning fat efficiently, not getting the right satiety signals, now, overeating and so on. So we cannot overstate the importance of short chain fatty acids like butyrate in the microbiome as it relates to obesity. So how does a dysfunctional microbiome then lead to weight gain and metabolic disease? Right, so what we've shown you so far is that there are critical components within the microbiome that support a healthier metabolic Response that support a healthier, leaner body. Higher levels of bifidobacteria functioning, lactobacilli, really important are increased levels of Akkermansia, increased production of short chain fatty acids. These are the critical factors in maintaining good metabolic health. But when the microbiome becomes dysfunctional, you start getting leaky gut, you start getting dysbiosis. And we'll talk about how that leads to a significant issue with metabolic distress. All of it really comes down to leaky gut. I mean, it's a primary cause of obesity and leaky gut, as you guys well know, is the non specific translocation of small molecules, bacterial components, viral components, even food components past the mucosa, past the intestinal epithelium and entering into basolateral circulation. One of the key things is this lps, this lipopolysaccharide making its way through a disrupted mucosal barrier. So LPS.

Ends up getting into the circulation through a dysfunctional barrier within the gut. Now, in a homeostatic environment, and some of you have seen this again, so it may be a review, but in a homeostatic environment, you've got nice, healthy, diverse microbes, lots of Akkermansia here, bifidobacteria, a huge diversity in microbes. They are not only maintaining an integrity among the ecosystem, but they're also maintaining the integrity of all of these structures of the mucosa, of the intestinal epithelium, of the tight junctions and so on. You get high production of short chain fatty acids in a good diverse microbiota. Those short chain fatty acids are keeping the inflammatory responses in the mucosa at a minimum. They are very powerful anti inflammatory compounds. The acetate that's being produced actually keeps certain overgrowing bacteria in check. It can act as a strong antimicrobial in the lumen itself. And it's one of the ways in which the microbiome checks itself. The butyrates of course, stimulating the GPR, GPR41, GPR43 on the adipocytes and in the intestinal lining as well. And of course, the microbes are also stimulating the production of and the expression of well formed tight junctions. So you're maintaining all of this homeostatic environment. The moment you start getting dysbiosis, whether it's because of a single course of antibiotics, whether it's because of poor eating for a period of time, whether it's stress related, whatever it is that's driving the dysbiosis, you start getting this degradation of the inner part of the mucus layer, which means you get lots of translocation of microbes, bacteria, viruses, viruses, bacteria, fungi, environmental toxins. All of the things that are up here in the lumen or in the top part of the mucosa starts translocating to the inner part of the mucosa and really close to the intestinal epithelium. Once it translocates close to the intestinal epithelium, you start recruiting in dendritic cells, macrophages, natural killer cells, cytotoxic T cells, to elicit really strong inflammatory response to protect the body from this influx of things that could potentially cause illness. And all of that inflammatory response starts breaking down the intestinal epithelium and the mucosa even further. It also completely dismantles the appropriate tolerance response to the immune system in the mucosa as well. All of the tolerance that your immune system is supposed to have is learned through mucosal sampling. All of that starts to fall apart. So then it becomes no surprise that your patients can develop things like food sensitivities and allergies as they are going deeper into metabolic dysfunction. So ultimately then what we're really concerned about is this moving across of lps, because lps, make no mistake about it, actually.

Initiates obesity and metabolic disease. So LPS creates this issue of metabolic endotoxemia, which is characterized by a sharp increase in serum lps. Postprandially, it tends to peak about five hours after a meal. Once the LPS gets into the system, it's followed by an activation of something called a toll like receptor 4 CD14 immune pathway. This activation is the biggest source of chronic low grade inflammation in the body. Metabolic endotoxemia is associated with dysbiotic microbiome and really caused by a dysfunctional barrier system that's not being maintained by microbes. And it's a self perpetuating problem, because when any given section of your gut lining becomes inflamed and becomes leaky, it causes more and more recruitment of inflammatory innate immune actors, which then creates adjacent areas to it, or leads adjacent areas to that leakiness to become leaky as well, because the inflammation keeps spreading down the line. So it's a self perpetuating problem if it's not corrected and managed. And we know that lipopolysaccharides in itself, as they get close to the intestinal epithelium, they themselves can increase intestinal tight junction permeability. So just having LPS moving through that mucosa, not having that really Important mucosal barrier allows LPS to move deeper into the lining of the gut, gets close to the intestinal epithelium and can create dysfunction in tight junction proteins. So they start dismantling the tight junction proteins through inflammatory responses.

As I mentioned before, metabolic endotoxemia, or the movement of LPS from the lumen into circulation and initiates obesity and insulin resistance. People aren't experiencing elevated levels of metabolic endotoxemia because they are obese or because they have metabolic dysfunction. They have metabolic dysfunction and they have obesity because they had increased levels of metabolic endotoxemia or intestinal permeability to begin with. And in fact, when you incubate LPS with Oedipocytes, you start to see the effect it has directly on adipocytes. So this is control wild type adipocytes, this is combining it with high fat. But then when you. And as you can see, there's no real change. Combining it with CD14, which is the expressed cytokine that increases the inflammatory response. Also, that doesn't have a direct effect. But then when you add LPS into wild type adipocytes, you see a swelling of the adipocytes. So as LPS interacts with the fat cells, it actually swells the volume of the fat cells by a couple of factors. And so that's one of the reasons you see this kind of swelling of the fat mass, especially the visceral fat mass and the fat that's around the midsection. The moment you eat and LPS enters circulation, it's going to go and interact with fat cells. So you'll get this massive swelling or ballooning of the fat cells. Now, what's scary about LPS is it also induces dysfunction in insulin response. And it does it through a couple ways. This is one of the scary ways where it can induce low grade inflammation in the hypothalamic JNK region and that causes something called central insulin resistance. And this has nothing to do with your body weight, just the fact that LPS can enter the brain and cause central insulin resistance, where it becomes really difficult for your brain to read your blood sugar levels. So managing blood sugar response becomes really hard and in fact can lead into type 2 diabetes. And again, this is part of the explanation for why we're seeing younger and younger people walking into doctor's offices presenting with type 2 diabetes even though they have normal body weight and their normal body composition. And so that is really a scary aspect of it, because we're driving insulin resistance in these patients as.

Their guts are allowing them to become obese as well, due to the endotoxemia. This is another really important study to take note of. It's called the Choreoprev study. What they were able to show, and this is following 462 patients over 60 months, they were looking at a number of of biomarkers that they could assess as predictive factors for the development of type 2 diabetes. So they took people who had increased risk for type 2 diabetes and they followed them, and they found only one thing, and that's high levels of postprandial endotoxemia. And they conclude that their results suggest that a high postprandial endotoxemia precedes the development of type 2 diabetes. They even suggest that LPS plasma levels can be a really important predictor of. Of the development of type 2 diabetes. So it's so important to remember that LPS moving itself into the circulatory system is the root cause or the root cause of so many of these issues of metabolic dysfunction, of obesity, of type 2 diabetes, and so on. And one of the things that also occurs is you end up getting this leptin resistance, because once LPS is in the bloodstream, it accumulates in areas like the hypothalamus, and it can trigger an inflammatory cascade in the brain itself. If LPS reaches the vagal afferent nerve, it can damage the nerve, resulting in a miscommunication between the gut and the brain, or the brain and the rest of the circulatory system, where you result in leptin and insulin resistance. So when you think about this, a lot of it is having an image impact on the brain and the microbiome when it's dysfunctional and it's driving the individual towards metabolic disease and obesity. So much of that is because of the significant impact it's having on the brain. Right? Inflammatory responses in the brain, inflammation in the hypothalamus, inflammation in other regions of the brain that actually disrupt the brain's ability to read what's happening in the body. And so your body needs to release more and more leptin, but it can't read the level of leptin. And so now you're getting this leptin resistance issue. And when leptin is not released and doesn't trigger the brain, then you get ghrelin, the hunger hormone that continues to escalate. So now you've got a continual expression of the hunger hormone and loss expression of the satiety hormone. Right. What does that do? That leads people to overeat now, here's the other connection. LPS endotoxemia also creates an issue with dopamine binding in the brain. So one of the things that LPS can do when it enters the brain is it interferes with dopamine receptors. Now, what's happening is people aren't getting that dopamine stoking that they need to satisfy the reward centers of their brain. So now they're dopamine deficient in response, and they have to do more and more extreme things to try to get that dopamine release. Things that lead to addictive behaviors, whether it is getting addicted to things like alcohol, to porn, to social media, to food, all of those things are trying to stoke and trigger dopamine release. And it's all the while the leakiness in the gut is stopping the dopamine from actually binding to its receptors in the brain. So so much of the dysfunction of the microbiome as it relates to obesity and metabolic disease has to do with how the microbiome impacts the brain in a significant way. So that cannot be overlooked. So microbiome obesity and metabolic disease. Metabolic endotoxemia induces obesity and diabetes. There's a number of studies on this, a number of them being done by the American Diabetic association and having published by the American Diabetic Association. So there's a big focus on how metabolic endotoxemia is the initial insult in the metabolic process that leads to type 2 diabetes and obesity. Increased circulating LPS causes both insulin resistance and then can also lead to things like leptin resistance and overexpression of ghrelin, ultimately things like dopamine issues as well. LPS causes morphological swelling of the adipocytes and. And it can disrupt the function of the enteric nervous system, which then causes, of course, misreading of leptin. It can slow down bowel movement. Right. So one of the things that tends to occur in obese individuals is they have increased energy harvesting from the food, meaning each meal, actually, they absorb somewhere around 200 to 300 more calories from each meal. So if you take a lean person and an obese person, you feed them the exact same meal, the obese person will absorb somewhere around 200 or so more calories from each of those meals compared to the lean person, even though they're eating the same exact thing over a period of 10, 15, 20 meals, that is enough of an increase in caloric intake to accumulate one pound of fat. Those little minute differences make a huge change over time to someone's body composition and metabolic response. Some of that energy harvesting is partly due to slowing down of the gastric emptying and the movement of the bowel, in part because one of the other effects that LPS has as it moves up the enteric nervous system is it gets into the dorsal vagal complex. When it enters the dorsal vagal complex, it stimulates an inflammatory response which starts with the activation of tnf, tumor necrosis factor. And when TNF is elevated, it stops the signal from the brain to the gut for peristalsis and for the migrating motor complex. So you start to slow down bowel movement and you start to let food sit in the small bowel longer. You get more aberrant growth of bacteria, you get more absorption of calories from that process, and you slow down overall bowel movement, which then has other complications as well. And remember that when you've got leaky gut in a single section of the gut, it is a self perpetuating problem because it causes other adjacent areas to be leaky as well. So this is just a schematic that talks about the multitude of different ways in which a dysbiotic gut microbiome, whether induced by diets or antibiotics, whatever the impact is, it can screw up the metabolic response in a number of ways. We talked about the short chain fatty acid production. One thing I mentioned earlier, GLP1, I did not mention peptide YY, but both of these are extremely important. These are bitter receptors. And as they pick up and actually combine with short chain fatty acids and bitters and other compounds within your food, they actually are triggered. And these actually improve insulin response in the body. So one of the targets for diabetic drugs right now is both peptide YY and GLP1. They find that GLP1 expression and peptide YY expression is attenuated in obese diabetic individuals. This of course can affect things like satiety with leptin imbalance and can lead to obesity and metabolic syndrome. The bile acid component, which I talked about a little bit, where bile activates the FXR nuclear receptor and it alters that release. And because it alters that release, it also can impact the TGR receptor 5 activation. Eventually. What that does is disrupts lipid and glucose homeostasis. Because you don't have adequate bile to absorb lipids for metabolism, you also then lead yourself into more leaky gut, more insulin resistance, and again leads to obesity and metabolic syndrome. Endotoxemia, we talked about quite a bit. So this just illustrates all of that for you in a simple diagram. One of the questions that we asked ourselves is can probiotics be used for metabolic health and weight management? And in fact there's a meta analysis and shows, and there's not a whole lot of studies on probiotics and metabolic health, but this meta analysis concluded that certain types of probiotics, they were very specific to strain level, showed that consumption could reduce weight and BMI significantly. But it was seen especially in overweight and obese individuals. And I know a number of the studies were also done in people with liver dysfunctions. So with people who already have early stages of non alcoholic fatty liver disease or even all the way till non alcoholic steatohepatitis, those individuals saw benefit from taking specific strains of probiotics. And like Lactobacillus rhamnosus GG as an example, or Bifidobacteria BLP1 was another species. And the effect lasted, was more profound when the administration lasted more than eight weeks. So there is some effect of certain probiotics right now on weight management. But we wanted to look at what would be the real profound change. Can we change that person's microbiome to look more like the microbiome of a lean individual, someone that has always been lean their whole life and never struggled with weight, One of those people that we always say can eat whatever they want, never gain weight, can we make their microbiome look more like that? And if we do, does that have an impact on how their body responds to foods that they're eating and any kind of food? Right. We're not even looking at diet food, we're looking at any kind of food, even the bad food food that they're eating, can it change how their body responds to it? So we looked at this as targeting, kind of the root cause of obesity and metabolic disease. And to us, the most powerful approach would be the use of a synbiotic, which is the perfect combination of a prebiotic and probiotic, where both the prebiotic and probiotic have complementary effects in the microbiome. So their effects are more additive rather than separate functions within the microbiome. So true synbiotic is defined really as a prebiotic that enhances the colonization of the probiotic. However, in a more expanded definition of a synbiotic, it's being developed that synbiotic is really a complementary prebiotic and probiotic that have the same effect within the system. So the way we looked at it is when you look at the microbiome and obesity, obesity is associated with low levels of bifidobacteria, low levels of Akkermansia, muciniphila, low levels of postbiotics like short chain fatty acids, clas and high levels of metabolic endotoxemia. This is the characteristic of a dysfunctional metabolic gut. And this is an obese individual. Most obese individuals that you measure will have these factors within them. So our simple solution was changing those things. Increasing endogenous levels of bifidobacteria, increasing endogenous levels of Akkermansa muciniphila, increasing short chain fatty acid production, and of course, reducing or alleviating metabolic endotoxemia or stopping LPS migration. So the first step we did is can we identify prebiotics that specifically increase bifidobacteria or bifidogenic prebiotics? And sure enough, we found that there's a couple of oligosaccharides that have good studies behind them that show that they are highly bifidogenic. In particular, one called Xos or Pretix is the name brand. It's a xylo oligosaccharide. And what they've shown is that even in doses as little as 1.4 grams, you see a measurable increase in the amount of bifidobacteria over the baseline. When you look at CFUs of Bifidobacteria and there was a little bit of a dose dependency when you went up to 2.8 grams. But even again at 1.4 grams, over time you see a significant increase in a 28 day period, it turned out that 1.4 2.8 grams did not make a big difference. So it's a cumulative effect over time, which is really promising to see. So that's really exciting. So we said, okay, that's something to keep in mind as part of the synbiotic. The other studies also show that xos, the xylosaccharide prebiotic, not only of course does it increase bifidobacteria over time, but it does have an impact on short chain fatty acid production. So when you look at the placebo group, there's really no change in short chain fatty acid production. But when you look at the group that was treated with Xylo oligosaccharides, you see a massive increase in short chain fatty acids, especially butyrate, which is really exciting because butyrate is the key one in metabolic function. So you see this is just showing you the same chart in a different way. Now you do see a decrease in acetic acid, and that may be because you're getting more acetic acid then being converted into propionic or butyric acid as well, which is not a bad thing. So the increase in butyrate was really exciting to see. And then you see overall short chain fatty acid production. And looking at the Xos, you see compared to other oligosaccharides.

Galactooligosaccharides, fructooligosaccharides, this mother's milk, if you will, you see a much higher increase in butyric acid versus other oligosaccharide sources. So that is really exciting because of course, butyric acid is critical in the weight loss segment. So we said, okay, we have the prebiotic, we know it's going to increase bifidobacteria, we know it's going to help increase short chain fatty acids. What about stopping metabolic endotoxemia? If you guys have seen us talk or are already familiar with what we do, you know that the spore based probiotics already stop the metabolic endotoxemia. So we know we can stop leaky gut from translocating. We also know that that has a profound impact and we've shown it through this type of study. But one of the most exciting impacts of that that we saw in that very same study is the impact on ghrelin. And this is exactly what we want to see, right? The hunger hormone ghrelin, when measured at baseline in the treatment individuals, what we saw is when they come in fasted at time zero, the ghrelin levels are elevated, as you would expect, which is totally normal. But then we gave them a 2000 calorie meal here and we barely see any drop in ghrelin at all. And then within a five hour period, it starts to increase, increase in fact. So we're not getting that ghrelin response, we're not getting that hunger hormone reducing quickly like it should. And in fact, we also tested leptin and we did not see the spike and increase in leptin like we should. Now after 30 days of taking the probiotic, the megaspore, we see this normal.

Basically re establishment of ghrelin function. This is what you want to see ghrelin do. The baseline ghrelin levels itself was less, right? And that actually reached significance as well. And what we found was that after a meal, after this fasted state, you take a meal, you see the steady decline in ghrelin, and we saw a steady increase in leptin as well. So just the spores alone were able to make that type of metabolic change by stopping leaky gut and reestablishing that gut brain communication between the spores and the prebiotic. We thought that's a significant impact already on a couple of the main factors associated with metabolic dysfunction. We decided to study them in combination as a true symbiotic to see if we saw that symbiotic effect. Sure enough, when we combine the spores and the prebiotic, we see a dramatic increase in things like butyrate production, in things like bifidobacteria growth, faecalum bacteria, which is another important keystone strain associated with inflammation in the bowel or inversely correlated with inflammation in the bowel, more importantly. And then we also see a significant Increase in Akkermansia, 10 to 100 fold increase in Akkermansia when you combine the prebiotic XOS and you combine the spore based probiotics with the megaspore. So this was exciting to us. And we then used the combination and in fact, in this study we actually didn't even use the entire formula of Megaspore. We just used the subtilis and the coagulates, just two of the strains within Megaspore, and we combined it with the xylosaccharide, the xos, to test and see the impact of this prebiotic probiotic mix over a 90 day period in overweight or obese women. So this is a pilot study and we're, we're looking now, once this study publishes, to produce a larger version of the study. But basically what we did is we took overweight or obese individuals and we put them on a probiotic prebiotic supplementation for a 90 day period with no change in diet, no change in activity at all. They are still consuming the type of diet that led to some of the obesity and weight gain in the first place. They're still eating fast food, they're not exercising, so they haven't altered any of the important behaviors at all. This is just looking at a fundamental change that a small intake in a single prebiotic and two probiotic spore strains can do in these individuals. So one of the first things that we saw that was super exciting was a significant decrease in visceral body fat. And this is measured through DEXA analysis or DEXA scanning, the dual X ray scan. And this is of course the most unhealthy location of body fat. All of the visceral fat in this truncal area, there was a huge decrease in visceral obesity. We actually saw about, in the placebo group, we saw an 11% increase in visceral obesity, which is not of course surprising because in these groups in this Whole cohort, we had them not change anything. They were still doing the diet the way they normally do. They still having a lack of exercise like normal. So it's not surprising that in placebo group they ended up continuing to gain a little bit of weight and gain visceral obesity. And yet in the treatment group, they had a 12% decrease in the amount of visceral fat mass. So that's a significant change in the body composition without changing anything about their lifestyle at all. So the microbiome alterations in itself has this kind of impact. We also saw things like total cholesterol dropping quite a bit in a measurable way, which is really quite exciting, because of course, high levels of total cholesterol are also associated with other risk that fall within the metabolic syndrome, like cardiovascular disease, for example. So that was a really promising thing. We saw a significant decrease in triglyceride levels. And we know that triglycerides are also associated with an impact on the liver. So when the liver is undergoing increased amounts of toxicity, one of its responses is the production of triglycerides. And so we know that the liver is being supported in this case as well. We also published a paper earlier this year in a trial using acetaminophen induced liver.

Cirrhosis or dysfunction that we found, or liver toxicity, not cirrhosis, but acetaminophen induced liver toxicity. We found that just using the spores alone actually protected the liver very significantly through acute damage, just as well as milk thistle did. And this is prescription grade milk thistle. So we know that the spores are protective of the liver and that has a huge impact in the metabolic health as well. We also saw significant reduction in C peptide, which is an index of the quantity of insulin your body is making. So we're seeing improved insulin response in the body, a reduction in the overproduction of insulin, which then of course can hopefully reduce the risk for insulin resistance. And then of course, currently we saw the reduction in the amount of fasting insulin being produced as well. So this was really promising also. And remember, these are individuals who are obese, overweight, who are not changing anything about their lifestyle. And we're seeing these fundamental responses within their metabolic response to food. And this is not the healthy food. They're not eating healthy food at all. They're still eating the same unhealthy food that leads to weight gain in general. So to us, that was extremely promising, even though it's a pilot study. But it really illustrates how much impact you can make on the way a person's body responds to food food just by starting to alter the microbiome. So imagine adding that kind of microbiome alteration into all the other things you do with lifestyle changes, dietary modifications, exercise programs, stress management, all of the other things you do. If you add this metabolic reprogramming in, the profound impact they're going to have on their results is going to be tremendously beneficial, of course to them. And then of course they're going to love you for it as well. So final thoughts on this that it's clear that certain probiotic strains can play a role. And it's really important to look at the right strains because one of the things that the probiotic studies showed on weight is there was highly strained and specific serotype specific. So it wasn't Lactobacillus rhamnosus that showed any effect, it was Lactobacillus rhamnosus GG. It wasn't Bifidobacteria, it was Bifidobacteria BLP1. Those are the specific strain IDs that actually had an impact. So not just any probiotic is going to have an impact. And different strains from different genres may have different mechanisms of action within the system. But we're very excited about the way the spores seem to impact metabolic activity. And more evidence is accumulating that you cannot achieve successful weight loss and metabolic health improvements without addressing the microbiome. To me, that's a huge missing link, not only in their ability to achieve weight loss, because you can achieve weight loss in a number of ways, right? You can be pretty radical about it. You can do crazy diets, you can do two a day workouts, you can do all kinds of things to achieve weight loss. But then the question is, can they keep it off or do they have to maintain this highly intense lifestyle to maintain that body weight? Or can we reprogram their microbiome and thereby their metabolic response to food so that they are one of those people that can be a little bit more lax about their lifestyle and diet and still not gain weight? Right. So that kind of change in reprogramming to me is really important when we're looking at long term solutions. To us, symbiotics will truly be the best solutions for faster, more realized weight management with the ability to fundamentally change how the body responds to fuel food. Our work on the xylosaccharide and our Bacillus endospores, it shows that we can repair some of the fundamental disruptions within the microbiome that is associated with weight gain and metabolic disease. Right. So there's a sustainable solution we're looking at here as a way of fundamentally changing how the body responds to food and then addressing dysbiosis. And leaky gut is going to be absolutely paramount to successful weight management. That leaky gut disrupts so many things. And not only is it, of course playing a role in the person's obesity, metabolic dysfunction, metabolic disease, it's also increasing their risk for all kinds of other conditions. If you've listened to my other lectures before, I show you evidence on how leaky gut impacts all kinds of other things, especially lps, endotoxemia, including mood disorders, autoimmune disorders and so on. So addressing this especially, we're doing it during metabolic revamping of a person's system. It really will help them overall in their overall health composition. So thank you again. I'm happy to take questions. And there's one more slide I wanted to put up about our Microbiome Keynotes. We were very fortunate to have Sachin speak to for us last year. He kind of kicked off the entire event, in fact, for us, on the first big night, and we were able to do that in person, which is wonderful. But this year, Microbiome Keynotes is going virtual.

This has been my passion of bridging what I call the translational gap. And in fact that's a. It's not my term, so I don't want to take credit for it. But the translational gap, that is a well understood issue between academic research and clinical practice. The vast majority of things that occur in academic research never come down to the ground for clinical practice, never impact anyone in a positive way. What we try to do as a company is bridge that gap. We try to bring the researchers, get them to mix and mingle with the clinicians and get the clinicians to understand how that research that they're doing can be utilized in a practical manner to impact the lives and overall health and wellness of your patients. So Microbiome Keynotes brings in some of the top microbiome researchers. Some of them are clinicians, most of them are researchers that they come in and they provide huge insights into the areas of research that they do within the microbiome. And we have very specific topics. Microbiome in your skin, for example, the vaginal microbiome, your plants microbiome, all kinds of things that are super exciting. I can't wait to hear most of those talks as well. So come join us at microbiome keynotes November 12th through 13th. Sorry, 13th through 15th. And we also have A setup where we do a lot of interaction. We have these mini masterminds where if you attend the event live, you'll actually get to spend time upwards of 20, 25 minutes with each of the speakers in special zoom rooms where you can actually interact with them casually and ask them all kinds of questions that you might have. So we want to bring and bridge that interaction quite a bit. So, Sachin, thank you so much, my friend, for this opportunity. If people have time, I am happy to take question.

Karan, thank you. I actually wrote a couple of things down that I wanted to discuss with you. Yeah, I'm just going to reclaim the host so I can turn my video on here. One thing that you said was about. And there's some questions that came into that I want to make sure we get to. So one thing you said was that when we lower lps, that seems to have a very positive effect and a cascade effect that creates a positive outcome. And so what is a dietary nutritional way to lower lps? Would it be to lower fat intake? Would that be the best way to do that?

So that can. So studies show that an increase in saturated fat intake increases LPS endotoxemia. And in fact, the most egregious of the fats that do that is actually coconut oil. So coconut oil increases LPS endotoxemia quite dramatically. So if you have overweight patients who are struggling with obesity, they shouldn't be taking in coconut oil, they shouldn't be doing spoonfuls of mct, because that can have a huge impact with metabolic LPS endotoxemia. That's one of the ways. Of course, increasing dietary fiber intake has a huge impact as well. That brings the short chain fatty acids up, which improves the barrier structure that prevents the LPS from translocating. Now, another reason why dietary fat intake can increase LPS endotoxemia is because one of the ways that your body absorbs fat is it produces these lipid rafts or Shiloh microns that go across the membrane, pick up dietary fats and bring them in. Because LPS is a fatty acid, has a huge fatty acid tail. These shilo microns actually grab LPS as well and bring them in. That's another reason why increased fat intake can have that impact.

Okay. Yeah, because one of the things that we do with our clients is we put them on a low fat diet and it gets them kind of scratching their heads a little bit because they're used to being on a ketogenic diet where it's a high amount of fat Right. Obviously that didn't work out for them. That's why they're, you know, consulting with us at this point. And I just wanted to make sure I clarified that. And that really helps explain why they see such dramatic results. But it's also promoting. It sounds like a healthier intestinal barrier as well.

It is, yeah. And in fact, the old adage of how fat makes you fat has definitely been debunked in many ways. Now, when we look at it at a molecular level, the understanding is that increase in fat intake, making you fat, is in large part driven by the increase in metabolic endotoxemia. So that's the molecular connection between intake of saturated fat or increase in saturated fat intake and then the resulting inflammatory metabolic dysfunctions. And there's a number of great papers on this showing that molecular connection. And it's not what we thought back in the 60s that it's been perpetuated over time. It's really this endotoxemia component of it. I had one more thing to say about that, which I forgot, but. Oh, and. Yes. So another thing that increased fat does is it reduces the amount of bifidobacteria and Akkermansia as well, because those colonic bacteria don't do as well on a diet that's of higher fat.

Okay. So I like that, that that really helps us understand kind of the mechanisms behind it and helps us lean into that a little bit more. And of course, for a variety of other reasons. You know, fat can be very hard to digest on our liver, and it takes a lot of metabolic energy to do that. And some people's livers and gallbladders aren't functioning at a high. High enough level to tolerate that level of fat. And when you say saturated fats, I know coconut oil is one example, but would you include meats and butter and things like that in that equation?

I would, yeah. I mean, it's. Coconut has a very unique composition of saturated fats, you know, the highest natural concentration. So some meats as well, certainly, you know, meats have somewhere around 40% or so saturated fat versus coconut oil, which is more like 80, 90%. So meats can be a source of that as well. That should be reduced, especially if you're already obese. Because, you know, another thing that I brought up, which you just mentioned, is this whole issue of the compromise of the liver in all of this. Right. Because the liver is already being compromised if you're obese. That's a given.

And it's only a matter of time that On a CT scan, you can actually see the fatty liver disease, but prior to that, prior to being able to visualize it on the CT scan, it's still happening in the liver. If you are overweight and if you have visceral body mass and all of that fat that's not being broken down, not being digested appropriately, not being absorbed and, and taken in by the liver through bile conjugation is going to end up as free fatty acids in circulation and that causes all kinds of problems.

Okay, thank you. Such great insights. So I kind of go through some of the questions that came in and for those of you that have questions, feel free to type them in. I want to be respectful of Karan's time as well. So we'll take a few and then we can, you know, pass these along to your team. I can save the chat if you want to respond to those. The other thing I wanted to know from you, Karan, is if you had a link that we could share for the Microbiome event.

Oh, yes, we can provide a link. Yeah, absolutely. I think the event has its own website. Microbiome Keynotes has its own website, so. But we will provide a link that you can share.

Absolutely. Okay, well, as you're answering this next question, I'll see if I can dig it up in one of the emails we sent.

All right, so Danielle's asking, what do you recommend for helping with the conversion of sulfur when the mechanism is compromised and some bacteria have taken over the task and cause hydrogen sulfide gas?

Yeah. So one of the key components will be, is reducing sulfate rich foods. So we, there are studies that show that over time, as you reduce sulfate rich food foods, you actually bring down the constellation of bacteria that are really high sulfate reducers that produce all of this hydrogen sulfide. So it's going to be a matter of, I'd say about six weeks or so of dramatically reducing those sulfur rich foods or sulfate rich foods. And many of them are healthy foods. Unfortunately, this number of vegetables, fish, tends to be higher in that, but just a period of about six weeks of reducing those will change the proportion of the sulfate reducing bacteria to things like methanogens, saccholytic bacteria, proteolytic bacteria, and so on. You can make those microbiome adjustments pretty quickly. So I would just say reduce the sulfate rich foods for a period of time and then you can start stepping them back into the system.

Okay, love it, thank you. And I just posted a link to the event in the chat, so feel free to click on that and it was an amazing event. I actually want to acknowledge how amazing the event was last year, and not because I had the privilege and the honor to be one of the keynote speakers, but because you guys did a really bang up job and the information was outstanding. And I know that this year it's online, but what I found is that I actually am enjoying these online events because you can connect with people in different ways. You know, for me, somebody who's an introvert, I can connect with people that I may not normally connect with. And so the technology has been amazing. And, you know, we're one of the vendors at the event and we're proud to help sponsor it and help put it together and be involved in it this year as well. And so one of the things I'm realizing is your team is doing, and I just wanted to acknowledge them as well, that they're doing such an amazing job ensuring that this event is going to be powerful and it's going to deliver tons of value and actionable steps that clinicians can take to accelerate their practice. And the cool thing about it too is whatever money they're going to spend on traveling there, or whatever money they were going to spend on their hotel, they can spend that money or invest that money into products. Right?

Exactly. Yeah.

And.

I'm excited to see what you guys have put together this year.

Yeah, thank you so much. And one of the things I always like to point out is that you will hear from people at this event that you won't hear from any other events that are put on. Because we really focus in on the highest level, microbiome researchers. And each of them specialize in very specific areas that are absolutely relevant to the health and wellness of your patient. You'll be learning from the best of the best in each specific area. And we're careful to curate and use people who are good at speaking. That's a problem when you work with academics and researchers. A lot of times their ability to convey the information is really.

Disrupted in some way because they're speaking at a certain level and they communicate in a way that really doesn't engage people. We try to do a good job of curating people not only who are tops of their field, but can also communicate the information effectively. But then you have all that time with each of the speakers within your own little private zoom rooms with a small group of other people so that you can really pick their brains.

I love it. I love it. It's worked out. Personally for me, I feel like it's worked out really, really well, going digital, of course, I miss hugging people and sharing their bacteria right when we're all together. But it is great to be able to connect with people online. So wishing you, and I'm going to be there, but wishing everyone who's tuned in today. If you loved what you heard tonight, then you're just going to be floored and blown away by what you're going to hear at the event. I was so pumped after I left there, so excited for how practical the information was. We had somebody asking if, and I'll kind of paraphrase what they're asking for context, but if you had to put somebody on a stack, a product stack for weight loss, to enhance weight loss, to maybe get, you know, some of the initial weight off or maybe the last few pounds off, what would you recommend? Where would you start, people?

You know, what we've done in the past, so we've done this to a certain degree. We don't see a whole lot of weight management people, but we have. And one of the things we've started them on is we just without changing any of the lifestyle components because oftentimes that's the hardest thing for them to change, right? They're so baked in how they live and how they eat and what they do each day. What we started to do is just put them on the prebiotic and probiotic mix without any expectation.

For a period of a month or two, saying that just start with this, we're going to start priming your gut for the next phase of your weight loss journey. What we tend to find is that just from that alone, many of these people, like we saw in our study, already start to see weight loss. They start to see a change in the amount that they're eating each day automatically because they're being reprogrammed. The gut and brain are being reprogrammed. And then they start to feel energy in a different way. And then they become really motivated for the next phase of changes. Right? Now, on top of that, if you wanted to, you can add things like, you know, l, carnitine and other things that do increase fat burn. You can add things like a little bit of caffeine for people to give them a little bit more metabolic stoking. You can make very small dietary changes in the beginning, maybe just have them eliminate one food or add in one little changes that we found when they start to see the effect with less expectation, that really motivates them. Because if they go on of a piece program in our experience, and we may not be creating Great programs. But if they go on a program and their expectation is, I'm going to lose £3 or £4 a week, and they don't achieve that kind of level, they get discouraged. The change is so radical for them that it's hard for them to manage that. So we start them slow. That way we explain to them that, hey, this is a little bit of reprogramming. Don't expect anything. Then they step on a scale and they go, wait a minute, I'm four pounds lighter and I didn't really do anything, you know, and then they feel more energy. That has been successful for us in the beginning. So that may be a way of thinking about the people who are more resistant to the change, getting them going, you know, stoking that motivation in them.

So automatic weight loss. Right. Without asking for too much.

Yeah. Without much commitment on their end, you know, without having to make too much adjustment. And, you know, as you guys know, you deal with a lot of people who are in this category or trying to lose weight. When they start seeing the results, that's when they really start getting motivated and excited. Right. And imagine they start to see results without really even doing anything. And then your next explanation is, hey, if you do these things on top of it, think about the response, right? And then that just gets them all jazzed up.

So what were those products then? What would you say you would start people on?

So all we do is we start them on the megaspore, the HU58, to give them that increased level of the bacillus subtilis. And then the Mega Pre. The mega PRE contains 1.2 grams of that xylo oligosaccharide. It also contains 800 milligrams of a fructooligosaccharide that specifically increases the growth of Akkermansia. So between those two products, that alone is enough. Now, if you want to give them, again, something to help with energy or increase fat burn, that can help, too. That can kind of stoke them a little bit. Their metabolic fire, if you will, stoke that a little bit. But just those two things of what we do.

Okay, so I'm going to ask a couple of questions that I think would be beneficial to the group as well. And then I do want to cover the topic of mouth breathing.

Ah, yes.

And we'll talk about that for a couple minutes, and then I'll let you get to your beautiful family.

Yeah, thank you.

So one question I have is about intermittent fasting. So tell me a little bit more about that in terms of how that can be leveraged or utilized or what your thoughts are on it for microbiome health.

Yeah. So intermittent fasting, to me, is one of the easiest things you can do to really improve your health. I know it's. For me, it's been a huge benefit, especially in the days prior to Covid, when I was traveling almost every week, couldn't eat right all the time, I couldn't exercise as much as I wanted to. Intermittent fasting was a huge help for me from a metabolic standpoint and an immune standpoint as well. One of the effects of intermittent fasting is an increase in Akkermansia, and you also see a huge increase in the diversity of the microbiome. So that's huge. And it actually, when Akkermansia increases, one of the things you get is more stimulation for the rebuilding of the mucosal layer. So you get a turning on of a gene called a MUT T gene. That's one of the effects that Akkermansia has. That stokes your endothelium or your epithelial lining to produce more mucin. So that is a real big benefit of that. Also, one of the effects of intermittent fasting is a huge increase in human growth hormone, which can improve metabolic response in the muscles itself and then improvement in blood glucose management. Right. So there's huge impacts that way as well. Now, from a strictly microbiome standpoint, one of the really interesting things that occurs when you provide your body a period of 12, 13, 14 hours of not feeding is you start to feed all kinds of secondary and tertiary metabolizers. So your microbiome is layered with different microbes that do different stages of metabolism of the food that comes in. Stage one is what we call the primary digesters or primary metabolizers. They break down the larger macromolecules, the carbohydrates, some of the proteins, even some of the fats. They break them down into smaller metabolites and they create other metabolites. Those metabolites then feed the secondary fermenters. The secondary fermenters now produce a whole array of important things like coenzyme Q10, like conjugated linoleic acids, like vitamins and hormones, things like serotonin and dopamine and so on. So all of those important secondary metabolites only get produced when the primary fermenters are not active, and the primary fermenters are active. Every time you keep feeding in food, into the system, so the microbiomes keep switching off unless you take A period of time where no food is coming in. The secondary and the tertiary digesters that produce all of these critical compounds for our function, for hormone balance and all of that, they actually don't get activated. So we need to go through a period of non feeding because the microbiome is known to be a diurnal system, meaning it has this circadian rhythm. And certain microbes are only active when there's no food coming into the system. So it's one of the most important things you can do for total comprehensive function of the microbiome.

I love that. Thank you. You just blew my mind.

It's crazy, right? It's so much to learn about this. I did a lecture recently where I spent three slides going through all of the primary secondary fermenters. And it's mind boggling when you start thinking about it. And one of the things we know is we've just scratched the surface. There are things that are happening in the deep recesses of the microbiome that we have no clue about. And by not allowing the microbiome to do what it does through periods of not eating, we are really disrupting all of those critical functions.

So this could really help explain metabolically from a micro biome perspective why eating, why less is more, why we could actually have more energy, more nutrient extraction, production, assimilation, by eating less.

Totally. See, we always have this kind of false equivalency of calories in and then calories that we need to burn, right? As if we are like an engine. As if it's just like the amount of gas you put in and how many miles you can drive. That's not how we work. We are far more complex than that. In fact, most of our cellular metabolic function comes from secondary and tertiary fermenters. I'll give you one example. Urolithins, the production of urolithin A. For example, the production of urolithin A, which comes from the digestion of polyphenols, doesn't occur until you have your secondary metabolizers active. And urolithin A is absolutely critical for mitochondrial function in our bodies. So the microbes are producing this, sending it into our circulation. It's getting absorbed by our cells, it enters our mitochondria and it actually increases the amount of ATP our mitochondria can produce. It actually increases the amount of energy our mitochondria can metabolize from breaking down carbohydrates and fats. Our ability to produce energy from the food that came in is actually dependent on a Complete, complete digestion of the food through all of the layers of the microbiome. Right. And so if we stop it, if we arrest it, then we're not getting all of these other downstream benefits.

Okay. Wow.

There's a lot to unpack from everything that you share today. But I think, putting it simply is that our microbiome is extremely important in so many different ways, and there's lots of different vantage points that we could approach this from. And giving it a break is important. And I guess when we're giving, giving, you know, eating a break, it's actually giving our microbiome the opportunity to do its job, to work.

Exactly. Right.

Yeah. So that probably increases metabolism too then, right?

Absolutely. Yeah. And it improves things like gut, brain connection, gut skin connection, gut liver access, all of those things, all of those axises get stoked because the microbiome is now producing things that each of these organ systems need. Right. On the brain, it's neurotransmitters and hormones. On the liver, it's liver supportive compounds that actually reduce oxidative stress on the liver. There's so many things happening through the microbiome that we don't even know about that we attenuate by not giving it time to do what it does.

Well, I love your humility. Right. I mean, there's so much that, you know, but you're saying that we've just scratched the surface. And to me, that that says a lot about who you are as a person. So I appreciate that. One thing I, I did want to jump into because I could probably talk to you all night.

Same here.

And I know, and I know you could, you could go on as well, but to be respectful of your time as well. Karan, I appreciate this. I did want to talk about mouth breathing, and you said something about the amount of bacteria that's in the mouth versus the amount of bacteria that's in the gut or the representation of it. And it blew my mind. So I want you to share what you shared with me, and then maybe we'll talk about the difference between mouth breathing and nose breathing. And that relates to weight loss and a whole bunch of other things as well.

Yeah. So the mouth is a very, very interesting reservoir. If you look at the combination between the mouth and the gut, between the two, they make up almost 54, 55% of all of the microbes in your body. And you look at the surface area of the mouth, you're like, it's a tiny organ compared to this 28 something feet that the gut has but when you look at the distribution of that 54, 55%, the mouth contains about 23% of it, and then the microbiome has the other 29 or so, 29 to 30%. So it's absolutely insane that the mouth has almost as much microbes, as many microbes as the entire digestive system does. So the mouth is loaded with microbes. It doesn't have as high species diversity as the gut does. But the number of microbial cells in the mouth is absolutely mind boggling. Right, so your mouth is a massive reservoir. Now also, most of the bacteria in the mouth are really designed to stay in the mouth. The moment they end up in any other part of the body, they cause significant problems. One of the reasons why we have this important gastric barrier is because we are continuously swallowing all of this bacteria from the mouth through saliva. And the stomach acid is supposed to kill them. What happens if stomach acid gets compromised while mouth bacteria end up in the small intestine and start to overgrow and cause sibo? The compromise of stomach acid is a type of sibo, small intestinal bacterial overgrowth. What happens when you have dental work and the mouth bacteria allowed to enter circulation? It causes heart disease and a number of other things. Dental work. The amount of dental procedures you have is directly associated with your risk for heart disease. The mouth is a filthy place and it has a lot of dysfunctional microbes that shouldn't be entering the rest of the body.

Yeah, I read a study and it was done in children and they found that children who are mouth breathers had a significant increase in ear infections and also upper respiratory, respiratory tract infections. And we were kind of calling the mouth the butt of the face. Right. And we should be breathing out from the mouth only as much as possible and then breathing in from the nose. So as a microbiologist, I wanted kind of your perspective on that.

Well, you know, when you think about the immune response in the respiratory pathway, right. The nose is such an important area because when you breathe into the nose and the air goes in through the sinus cavities and the rest of the upper respiratory tract, that's where all of these cilia are. That's where you've got a mucosal layer that actually protects. That's where you've got all of the secretory IGA constantly being released as a way of understanding the viruses, the bacteria and all that come in through from the natural world into your system that could potentially cause harm. And we can elicit an immune response against it. When you're breathing through your mouth and the potential of inhaling, if you will, a number of those bacteria that may screw up the immune response within the system because those bacteria are already engaging with the buccal or buccal immune tissue in the mouth and that's creating a different type of immune response than what you want to see in your upper respiratory tract or in your sinuses, in your ears and so on. So I could see that being a significant issue. The fact that you maintain, you mentioned that to me is probably going to put me down a rabbit hole in this world as well, because that's really interesting. I mean, you could see physiologically how breathing in all of these crazy microbes and their endotoxins and all that that are produced in the mouth because you have a lot of gram negative bacteria in the mouth, they produce endotoxins, how that can be detrimental to immune response in the upper respiratory tract and so on.

Yeah, it's been fascinating. I've been reading the book Breath by James Nestor and the information I'm learning, it's something that has been kind of in my subconscious, if you will, because I know about pranayama and breathwork and things like that. And now I'm becoming a breathwork instructor. So I'm actually going through that training process and it's been this just amazingly mind blowing rabbit hole that I've gone down and it's starting to really make a lot of sense. So for, for the entertainment of everyone that's here, I'll, I'll give this a few minutes and I think it could be really valuable in connecting a lot of what we talked about today. Yeah, the diaphragm is referred to as the second heart. And what it's designed to do, it's designed to obviously when it contracts, pull the lungs down and create a vacuum. But at that same time it's also kind of gently squeezing the digestive organs as well and acting as a physical pump. The diaphragm is also referred to as the horizon. So it's the horizon between kind of the, the dirt, if you will, and then the lungs and the heart and those organs. When the diaphragm is working correctly, which as we know is one of the most innervated muscles by the vagus nerve. So when the vagus nerve is working correctly, the diaphragm is working correctly, and when the vagus nerve is working correctly, we make adequate amounts of stomach acid. When we don't breathe through our diaphragm, we actually properly, we actually make the heart work harder so it increases blood pressure because now we're not getting the squeezing effect, if you will, from the diaphragm. And so now the heart has to compensate for that. And people who are generally tend to be just in general, people don't have awareness of their breath, tend to be shallow breathers. And shallow breathing creates more of a sympathetic dominance in the body, which then creates even less stomach acid production, less, you know, immune function as well. So it. And then if you breathe into your mouth now, you're inhaling all this bacteria and further, you know, compromising the immune system. So it's all, you know, interconnected with one another. And it's amazing how simple things like, you know, being focused on your breath, intentional about your breath, breathing through the nose as much as possible, mouth taping, things like that, incorporating some fasting and then putting in some prebiotics and some spore based probiotics. I mean, that can create such a rapid transformation for people in a very short period of time without having a whole bunch of interventions. We're really taking two important aspects of what we do every single day, which is digest our food and digest our air, if you will. And those two things are often overlooked. And I think we can simplify health by really honing in on those opportunities that don't require a lot of expensive interventions or even testing. In some cases, just doing these foundational basic things can make such a huge difference for people. So thank you for those additional insights that you gave.

Yeah. Now that you mentioned that, there's two points I want to bring up to what you said that I think is super interesting. One is the breath work with blood pressure. Right. And that's a real effect because there's a device that you can buy that has a whole bunch of clinical trials and is an approved medical device for dramatically lowering blood pressure, in fact more dramatically than you can with medication. And then all it is is a breathing device. It's a device that helps you create a pace to your breathing. The device you actually stick it on, I think you stick it on your chest and then you follow the tones of the device and you breathe in and out. With the device. You do it like 30 minutes for a day or something like that, and it changes the depth and the pace of your breathing. And that alone over a three week period of time is shown to bring down blood pressure very significantly. And then the other thing you mentioned that is also really interesting as it connects to the microbiome is this vagal tonality. Right. So whether you are more sympathetic or parasympathetic. One of the aspects of a sympathetic kind of dominant tonality is that opportunistic microbes tend to think thrive in that sympathetic state. Because when the moment you release things like cortisol, epinephrine, norepinephrine, there's a number of microbes that live in our system that are looking for an increase in those responders. And the moment they see an increase in cortisol or epinephrine, norepinephrine, they actually increase surveillance factors because their assumption is the host is compromised. Now in fact, that kind of disruption of tonality can cause dysbiosis in the system. And then that increase in growth in opportunistic viruses and bacteria will lead to inflammation, leaky gut, and then the whole cascade of things. So just the breathing aspect alone that you bring up is so powerful. I'm glad you mentioned that because it ties in so well to what can happen in the microbiome.

Love it. Love it. One question coming in and then I have one more question we'll wrap things up. Is what is the name of the breathing device? Do you recall what the name is?

I don't. I need to look it up. But if you google. I found it because one of my friends was experiencing high blood pressure and was asking me if he should take medication. I was like, wait, no, no, no, because there's other ways of doing it first. Right. And so I googled blood pressure solutions or natural blood pressure device, because I remember hearing about the device. If you Google natural blood pressure device, you will find it. And it's got a 510 medical device approval by the FDA. It's something you can just buy off of the Internet and bring it home. It starts with an A, like a rest or something like that.

But respirate is that.

Oh maybe, yeah, maybe something with the rest in the word in it. But if you google it, you'll find it. That's, it's, it's advertised all over the place.

Lovely. And then I had a pressing question. I wanted to ask it earlier since I'm hosting you today. I'm going to ask.

And then we'll let you go. Is the fasting part, is there a recommendation? I know the intermittent fasting, the 16:8 is great, but is there times or intervals that you recommend? 24 hour fast or 3 day fasts that could help us go even deeper into cleaning up.

So based on the studies on the microbiome, there is diminishing return after a certain period of time. Right. So to me, the longest I've ever done is 24 hours. Beyond that, to me, I think we need to feed the microbiome a little bit. I've never done like a six, seven day water fast. I think that may have more spiritual cleansing kind of effect than real impact on the microbiome, for example. But I think an occasional 24 hour fast is perfectly fine and could be beneficial. But that intermittent level is where we see all of the magic happen.

Naturally. I think that's kind of more what our ancestors did and they didn't do it purposefully. They had no choice, they just didn't have enough food around to eat all the time. And then as far as the timing of fasting, there's some interesting studies that show that there's a slightly beneficial impact on metabolism if you're feeding windows during the early part of the day where you're eating from somewhere around 8am to little afternoon and then you're skipping dinner. But for me that's impossible to do. Dinner is too much of an event, both in my house, socially, business things and all that. So for me it's much easier to skip breakfast and do it that way. But that also gives you a huge amount of advantage.

Okay, so if people want to go a little bit deeper than they can eat in the first half of the day as opposed to the second half.

Okay. Especially for metabolic benefit. That's what they showed, that the weight loss was a little bit more.

Rapid when they fasted in the later part of the day and eight in the earlier part.

Okay, great. I guess you could always give people the option depending on their social patterns, work patterns, stuff like that. Totally amazing. Well, Karan, this has been such a blessing. As always, appreciate you appreciate all the work that you're all doing at Microbiome Labs. You have an amazing team, you've got an amazing event coming up and you know, I'm so looking forward to it. Thank you once again and my friend, thanks for sticking around a little bit longer and answering all of our questions. We really appreciate that.

It was a pleasure. Look forward to doing it again and can't wait to see you on keynotes.

Sounds great, man. Talk to you soon.

Bye. Bye.

Bye.