The Peter Attia Drive - #283 ‒ Gut health & the microbiome: improving and maintaining the microbiome, probiotics, prebiotics, innovative treatments, and more | Colleen Cutcliffe, Ph.D.

Episode Date: December 18, 2023

View the Show Notes Page for This Episode Become a Member to Receive Exclusive Content Sign Up to Receive Peter’s Weekly Newsletter Colleen Cutcliffe is an expert in molecular biology and co-founder of Pendulum Therapeutics, a company working to develop treatments for a variety of diseases by targeting the microbiome. In this episode, Colleen delves into the complexity of the microbiome, how it is tested, and how it changes over time. She explores how probiotics, prebiotics, and postbiotics affect the gut and makes a compelling case that well-developed products have the potential not only to enhance gut health but also to positively influence overall metabolic well-being. Colleen emphasizes the significance of a high-fiber diet in sustaining a thriving gut microbiome, shares insights on minimizing microbiome damage during antibiotic use, provides tips for fostering and preserving a healthy gut, and much more. We discuss: Colleen’s background and current focus [4:45]; The basics of the microbiome [7:15]; The study of the human microbiome [15:15]; Categories of bacteria, and the implications on health of the rapid evolution of bacteria [19:45]; Methods for measuring and understanding the microbiome, and key indicators of microbiome health [28:30]; The important role of fiber for promoting gut health through the production of butyrate [38:30]; The case for manipulating gut bacteria via fecal microbiota transplant (FMT) [45:00]; Dynamics of the microbiome: the gut-brain connection and how antibiotics, nutrition, stress, and more impact the microbiome's diversity and function [50:15]; Factors that influence the vaginal microbiome [55:15]; The effect of gut microbes on obesity and challenges with fecal transplants in people [58:45]; Beneficial strains of gut bacteria and strains commonly found in probiotics [1:01:15]; The difference between a probiotic and prebiotic, and how CFUs are a measure of the “active ingredient” [1:09:45]; Considerations about how probiotic strains are produced, and more on the meaning of CFU [1:14:15]; Mitigating the effect of antibiotics on the microbiome [1:22:30]; What do we know about the effect of artificial sweeteners on the gut microbiome? [1:30:00]; Why Akkermansia is a keystone strain with implications for metabolic health and an individual’s response to dietary interventions [1:36:15]; The essential steps necessary to develop a robust probiotic for optimal health support [1:45:45]; How Akkermansia helps control blood glucose, and potential implications of Akkermansia in weight loss, diabetes management, and more [1:48:45]; Pendulum Therapeutics’ commitment to rigorous product develop [2:06:30]; Details about the probiotic “Glucose Control” and other probiotics developed by Pendulum Therapeutics [2:13:00]; Further studies of Akkermansia that have been proposed or are underway [2:20:30]; and More. Connect With Peter on Twitter, Instagram, Facebook and YouTube

Hey everyone, welcome to the Drive Podcast. I'm your host Peter Atia. This podcast, my website, and my weekly newsletter all focus on the goal of translating the science of longevity into something accessible for everyone. Our goal is to provide the best content in health and wellness, and we've established a great team of analysts to make this happen. It is extremely important to me to provide all of this content without relying on paid ads. To do this, our work is made entirely possible by our members, and in return, we offer exclusive member-only content and benefits above and beyond what is available for free.

If you want to take your knowledge of this space to the next level, it's our goal to ensure members get back much more than the price of the subscription. If you want to learn more about the benefits of our premium membership, head over to peteratia-md.com forward slash subscribe. dot com forward slash subscribe. But guess this week is Colleen Cutcliffe. Colleen received her bachelor's degree in biochemistry from Wellesley College and a PhD in biochemistry and molecular biology from Johns Hopkins University.

She then completed postdoctoral training at Northwestern Children's Memorial Hospital and subsequently began working as a scientist in the pharmaceutical industry. She's currently the CEO and co-founder of Pendulum Therapeutics Startup that is working to develop treatments for a variety of diseases by targeting the microbiome. Although given that Pendulum has been around for about a decade, it seems a little odd

to refer to them still as a startup. I wanted to have Colleen on the podcast because quite frankly, she was the first person I met and had deep discussions about the microbiome where I really felt like the person knew what they were talking about. Now, I don't say that to be disparaging of the many other people who have a lot to say about the microbiome, but my mind works in a particular way. And I guess I've just never had that connection with a person where when I ask questions, they seem to have answers that make sense to me.

So much of the work that I've seen around the microbiome has been interesting, but it's been very difficult for me to understand how one could operationalize and make real causal effect from the science that is being presented. And so over the course of many months, Colleen and I had a number of discussions. I became quite interested in some of the products that her company sold. And I even began to see some of the benefits in my own blood work, something that I was incredibly skeptical of at the outset. In fact, we realized after our first meeting that many people had been telling me

about some of these products over the previous four or five years, and I had been generally quite dismissive without looking more deeply at the data behind them. In this conversation, we really dive into all things related to the microbiome. We talk about what it is and how it changes over time and how you can measure it if that is something that is important and we'll talk about the importance or lack thereof. We talk about probiotics versus prebiotics versus

postbiotics and how much bacteria actually make it into various products that one can buy. We speak about fecal transplants and the importance of fiber as well as artificial sweeteners and antibiotics as it relates to the gut microbiome. We then speak specifically about acrimansia and Colleen's work at Pendulum around creating products that focus on not just acrimansia, but creating substrate for it and other tools that will enable the gut to be fed in the best possible way to improve metabolic health. This is a really interesting discussion on many levels. And again, I just want to stress how skeptical I have been of this topic for at least a decade. In fact, there is probably no area of medicine that I have been more skeptical of than this one.

And of course, with some of the high-profile failures of companies in this space, I have somewhat smuggling felt vindicated in my skepticism. But I must say that I feel like that is changing, and I think that the work that Colleen and her team have shared with me, along with work that has been shared by other people, have began to make me think that there may indeed be something to this gut biome. In other words, it's not that I don't think that the gut biome matters, but the question is, can we change it

using tools other than our nutrition? And to me, that's one of the most important questions we dive into in this discussion. So without further delay, please enjoy my conversation with Colleen Cutcliffe. Hey Colleen, thank you so much for making the trip out to Austin to sit down. It's always more fun to do these in person. Thanks for having me. We're going to take advantage of being in Austin.

Yeah, I know. This is a topic that we've received a lot of interest on and people have always wanted to go deeper on this topic. And my reluctance to do so has been, in part, driven by a couple of things. One is just my general lack of clarity around finding people that can speak about the topic with some degree of rigor. And then secondly, it's just it's a market of products that seems so sketchy. And so when you and I met, it's probably been six months ago and connected,

we went for this walk and I was thinking, ah, this is like the most I've learned about this and the rest is history and so here we are. Maybe give folks a bit of a sense of your background scientifically. So I think when we met, we figured out pretty quickly, we had both been at Hopkins at almost about the same time that you of course were doing your PhD there. So what did you do your PhD in again?

Yeah, my PhD was in biochemistry and molecular biology. So really thinking about enzymes and pathways and how they all interact with each other. And then I did a postdoc at Northwestern. We were looking for diagnostic markers for pediatric Wilms tumors. And then I moved out to the bear and I worked in pharma, we were developing drugs for Parkinson's disease. I sort of followed a pretty traditional path of a scientist into pharmaceutical drug development.

Then I did what everybody does in Silicon Valley, joined a startup company, and this was a DNA sequencing instrument company. And we went through rapid growth, we went public, and on the other side of that I started this company, Pendulum, and it was really premised in the fact that things like probiotics and yogurts have been on the shelves for decades, but there actually hasn't been a new ingredient introduced in over 50 years. And that's because microbiome science wasn't a real science until DNA sequencing technology enabled us to really be able to survey the microbiome and create these metabolic maps and start to approach it like a biochemical

problem and a systems biology problem. And all of a sudden it seemed like, wow, there have been a lot of unlocks in DNA sequencing technologies that would allow this entirely nascent field of science and medicine called the microbiome to produce real products. And so here we are 10 years later in it. Well, definitely spend some time talking about products and the evolution that you've had in that path. But let's talk more broadly just kind of about this field.

First of all, how do we define the microbiome? The microbiome is essentially all of the microbes. So the bacteria, the viruses, fungi, yeast that reside in and on us, and they are on our skin, they're in our nasal passages, they're in our lungs, and they're in our guts. Got it. Now, our guts, which run mouth to anus, are outside of our body. So people don't think of that always that way, but of course they are. What allows the colonization of that? I mean, is that something that is set at birth? Maybe taking us to back when a child is in his mother's or her mother's womb? There's amniotic fluid that's flowing through that spot.

Is that a sterile area? Well, interestingly, for a long time, as you know, we've all believed that was an entirely sterile environment, and there were no microbes there at all. And some recent studies have started to elucidate that there are some strains, but it's very minimal. When we think about the gut microbiome of an adult versus somebody who's in the womb, I mean, it's incredibly much more diverse once you become an adult. And in fact, really the primary initial seeding of the microbiome is through the delivery

in the vaginal canal. And so we're going to get gross for a second, but literally as you're being delivered, you are consuming fecal matter that is in the vaginal canal. And that's your first seeding of microbes. And infants have a very small diversity of microbes that are really tied to mother's breast milk. And then as you start to eat foods,

and as you start to get exposure of environments, then the diversity of microbiome start to eat foods and as you start to get exposure of environments, then the diversity of microbiome starts to really grow and flourish. And then at some point on the aging process, the opposite starts happening. You start to become less diverse in your microbiome as we age. So you start out almost like a blank slate, you get a lot more diverse. And then as we age, you start to lose that diversity and therefore some key functions in the microbiome. When is peak diversity approximately approximately what decade of life?

Obviously very some person to person, but if you can remember a time where you could eat or drink whatever you wanted to and you didn't have to worry about it, that would probably be the time. Oh, so that's actually quite young. Teenagers. As a teenager, exactly. Yeah. When I could indeed eat a bowl of cereal for every meal with no consequence.

Yep. By bowl, I mean, a bowl the size of my head. So it's a box per bowl per meal. Okay, so we have a relatively early peak in life for diversity. You hear all of these sort of bumper stickers, slogans about the gut biome. Oh, it outnumbers us 10 to one. Is that true? I think those numbers have definitely come into question. I mean, they're nice to give people a framework for the fact that you have a ton of microbes

in you. And I think that's the important part is that they are, whether the fact that you have a ton of microbes in you. And I think that's the important part is that they are, whether they outnumber you tend to one or two to one, I think is relatively probably not that important. But what is important is that they make up a huge portion of your body mass as well as functions. And so it's an important key part of quality, even one to one. Yeah, even one to one is huge. So the idea is there are many cells that are not you between your mouth and your anus as there are you.

Exactly. Now, obviously, just to have someone wrap their head around that, we're made up 70% of water. So most of our mass is water, not the cells minus the water. Are these largely anhydrous cells? Like how do they weigh so little relative to the rest of us? No, man. I don't know the water content of bacteria, but maybe I think about it a little bit differently, which is more about the biochemist or everything's going to come back to that. But it's more about the biochemical functions, what's the output of each of these cells versus the output of our cells?

And I think when you look at it that way, these are real workhorses. So there's definitely redundancy among bacterial cells, but each of them is having multiple functions and multiple outputs. And so when you think about it at a cellular level, I would think more about what are the things being produced by the cell? And bacteria tend to secrete a lot of things that they're producing unlike the cells in our body. And so there's a lot of function that's associated with the microbiome that's super important.

So I remember in my first and only biology course as a kid, because I didn't take bio again until I decided to go to medical school in ninth grade or whatever, we learned about pro-carriots and eukaryotes. And I still remember to this day the joke of our teacher, Mr. Jefferson, he said, do you know what a eukaryot is?

And everyone was like, no, and he goes, it's a portaging term. You know, portaging when you carry canoes back and forth between rivers. Anyway, no, so a horrible joke, but I still remember it. So I'm going to have to cut this part out of the podcast. It's so bad. But where are we going? Where I'm going with this is, can you explain to people listening what the difference is between our cells and bacterial cells. Because there are some fundamental differences between these things called pro-carriers and eukaryotes. Yeah, I mean, I think that first of all, every cell in our body needs the other cells and the

organs in the whole system in order to be able to survive and do their job. Whereas bacteria, they don't need anybody else. And so every bacterial cell, every unit, is its own living thing that can replicate, perform functions, lose functions, be genetically modified, and all of that. And so it's sort of its own entity, its own living organism, every cell is a living organism. And then they divide really, really rapidly, some of them, as quickly as 10 to 15 minutes, you're dividing. And so there's this other component which is that some of these bacterial strains because they divide so quickly and because they're also under the pressures of evolutionary processes, is that they can evolve super quickly.

And so we as humans, we have a long evolutionary timeline because it comes in the form of you make a kid who makes a kid who makes a kid. Now imagine that was happening every 10 minutes. You can evolve really, really rapidly. And so that's part of the antibiotic crisis out there, which is to say that these things can become resistant to antibiotics because of this division time. You alluded to this already,

but they have the capacity to secrete things significantly. We think of bacteria and we hear it as a bad term. We think of a bacteria as a bad term. We think of a bacteria is a bad thing, and there clearly are some bad bacteria. But would I be oversimplifying if I said that most bacteria enjoy a kind of complementary relationship with us as their host? Is that fair in terms of flora, such as the bacteria on our skin, inner nasal passages in our gut? We've co-evolved with these microbes in these bacteria,

and so generally speaking, when you're co-evolving with something, there's some mutual benefit. I sort of cringe when people talk about good bacteria and bad bacteria, although I do it as well, because it's the ecosystem and the context of these bacteria that's actually more relevant. A good bacteria can become a bad bacteria in a certain situation, and likewise, good bacteria can become a bad bacteria in a certain situation, and likewise,

bad bacteria can become beneficial in a different context. And so I think that it's important to know that they're all part of these different pathways and what they're doing together. So, for example, Clistridium difficile is something that I think people think is a terrible pathogen, and it's so bad for you. And oh my gosh, you better never get it. Almost all of us have Clistridium difficile in our guts, but at the level that it's at and in the context of the ecosystem of our strains, it's not having that kind of really nasty pathogenic

impact in my opinion. They're really bad bugs. People hear me rail about good cholesterol and bad cholesterol being meaningless terms. And of course, cholesterol is simply cholesterol. It's where it ends up that can be good or bad, so that's actually a great analogy. While we're on the topic of Cluster to Immunificell, which we'll come back to in more detail, what is the prevalence of that as a function of total gut biome in a person who's healthy and not otherwise in a pathologic state?

Well, this is the convoluting part about the microbiome, which is that if you sequence and do really deep sequencing and even biochemical assays across a person's microbiome, and then you say, right now, I want to do population studies, what you find is that the difference between people is huge. And so the human microbiome project in which they can look across 10,000 plus people at all ages and different demographics really demonstrated that at the strain level, people are pretty different from person to person. When you start to look at the functions, that's where you start to see some redundancy.

So it's hard to say for a particular strain, if someone gives you an actual number and they don't give you a range, that's probably not correct. Wow, interesting. Tell me more about that project. So what were the observations it landed at vis-a-vis how various factors, both modifiable and unmodifiable, either genetic or age and diet being the most obvious modifiable factor? How did that impact the gut biome in these 10,000 people? Those

differences magnify in the outcomes. That initiative didn't have a longitudinal component to it or a perturbation of the system and looking at before and after. So it really was just a one-time observation. One-time observation and saying, okay, we just look across a population of people. And this is super early on.

So we didn't know anything about the microbiome. We barely knew how to sequence the thing. And so, and even things like, well, what should the sample be? Should it be a scoop of somebody's stool? Should it be an entirety of the stool? Should you do 16S sequencing, which is just a gene that all microbes have? Or should you do the whole genome sequencing?

That's going to be a lot more expensive. This is a government-funded project. And so there was a lot of unknowns at that time. So even just getting this information of, if I looked across 10,000 people from skin to guts to vaginal microbiome, what does it look like? That was a huge endeavor. And of course, coming out of that, it had been a ton of longitudinal studies and studies where people have done actual interventions. Going back to what you said earlier, it's not just bacteria, it's viruses, yeast, fungi. What does the pie chart of that look like in terms of numbers?

And then what does the pie chart like that look like in terms of function? I'm guessing the bacteria are doing the majority of the work. I do feel pretty comfortable saying viruses are bad. Is there an exception to that rule? I mean, we certainly can harness viruses to do good things for us in terms of recombinant DNA technology. But if there was not a single virus on this planet afflicting humans, I think we'd be in a better place, right?

I'm not aware of good viruses. Well, this field is really nascent. I hesitate to even answer the question of what is the role or the significance of all these different types of microbes. Interestingly, I think we also don't really know what the role of viruses are in the microbiome, but one could imagine that you have functions that are important and that

you need to be healthy and that maybe these viruses help accelerate movement of those particular genes from one bacteria to another. So I wouldn't say the jury's totally out in my mind. Interesting. to another. So I wouldn't say the jury's totally out of my mind on that. Yeah, that's a good point. Do we know anything about how our microbiome compares in complexity to that of other animals? A lot of microbiome studies are done in mouse models. And I mean, having been in pharma, I feel like if you're a mouse and you got cancer, you're made. We have so many different gears for you. The translation of that into humans is not that great. And the microbiome is even worse because animals eat different foods. Your diet is one of the biggest things that impacts your microbiome. And so now you're saying what is an animal's microbiome? How does it interact with the host? And then what implications that I have for

health? So you're even one more step removed. So there are a lot of strains that exist from mouse to man, but I think again, because it's context, it's not totally clear that you can really use these animal models to predict what will happen in a human. Has anybody looked at animals that are not living in captivity and not genetically ridiculous, like the typical black six mouse and things like that? I mean, do we know what our pet dogs look like or what animals in the wild look like? Do they more closely resemble us? Or again, is it purely a function of what they're eating?

A function of what they're eating is such a big driving factor. So if you look at your pet dog, I mean, it really does depend on what you're feeding them. Some people feed their dogs pretty much a grain-based diet. Some people are cooking chicken for their dogs every night. You know, so when they look at pets, I think that's also hard because it just ends up being parsed out by what you're feeding them.

Just to get back to the reality of these genetically modified mouse models, it's even more extreme in microbiome studies. So they literally make, I call them, like the bubble boy of mice. So they try to entirely deplete these mice of microbiomes. So they're taking a mouse that has no microbiome, and then they're infusing them with a human microbiome and they're saying, okay, what happens now to the mouse?

And so these are germ-free mouse models that are really commonly used to understand the impact of a human microbiome on the health of the animal. This is completely not, but would ever happen in real life. So for folks listening again, who might not think much about bacteria, my vague recollection of microbiology is one line in the sand we draw to divide them is, are they aerobic? Are they anaerobic? Are they gram positive? Are they gram negative? Which is simply a staining technique. I guess I should explain that aerobic are bacteria that require oxygen for respiration and aerobic are animals that generate ATP without oxygen. And then you have

facultative of each where they prefer to do it one way, but they can do it the other way. Any other divisions worth talking about as we explain the types of bacteria? The aerobic versus anaerobic is definitely one of the most important things you think about. You've got bacteria on your skin that definitely loves oxygen. And then what we call the gut microbiome is actually a strictly anaerobic area of the distal colon. So there's literally no oxygen. Those strains can't even grow in the presence of oxygen. One other thing might be localization. So your GI tract and the gut microbiome, there's this so-called

gut lining. So the cells, the epithelial cells, where there's this mucin layer, that's an important part of having a well fortified gut lining. There are strains that live in that mucin layer, and so that's a different type of a strain they feed off of mucin versus many of the strains that sort of feed off of things like the foods that you eat are prebiotics. How does the characteristic, let's just focus on the bacteria, how does the characteristic of the bacteria change from mouth to anus? Obviously, you're going into less and less oxygen as you go down.

So presumably, you don't have pure anaerobes in the mouth, but I know from the little bit I remember about working in an ER, whenever somebody received a bite, you would think, ah, how bad can a bite be? We were really conditioned to remember that those are some of the dirtiest wounds a human can have. No less dirty than a feces-soiled wound. So even at the approximate end of that bacterial lining, these are really frightening bacteria. Can you tell me anything about how the bacteria change as you progress along the length of that?

By the way, that's a very long tube. It's not just measuring here to here. Like the listener would have to understand how tortuous the small intestine and the colon can be. The primary thing that changes is this anaerobic part. So obviously your mouth, there's a lot of oxygen exposure. And then as we said, when you get to the distal colon, there's no oxygen there.

So all along that path, from the mouth to the stomach, I think is where you have a reasonable amount of oxygen exposure. Once something gets through the stomach, there's sort of this in between the strict anaerobic and the aerobic area. And a lot of the lactobacillus and bifidobacterium strains that are on the labels of probiotics out there today, they kind of reside in that small intestine area. And then

you get to the recesses of the gut microbiome, which is where all the action happens. So after your stomach has broken down foods and they make their way to the distal colon, that's where an incredible amount of metabolism is happening. And there, there's no oxygen. But yeah, the mouth microbiome is really interesting opportunity because what's happening in the mouth, the two ends that you mentioned, mouth and anus, what's happening in the mouth, and what's coming out the other side, give you an indication of what's happening in the middle.

I just tell you a funny story. Early on when I started this company, I met a guy who was really interested in hyena mouth microbiomes. The reason is because the hyena is one of these bizarre animals that can eat a carcass of an animal after it's been dead for extended periods of time, you know, week two weeks. Know that their animal go near an animal that's been dead for that amount of time because of the bacterial overload. This is just going to kill you.

And so it's so called rotten meat that these hyenas are able to eat and see this big question of why are they able to do that. And it turns out if you look at the microbiome of a hyena, they make an incredible amount of antibiotics. And so he had this whole theory that if you could understand what antibiotics are being generated in that hyena's mouth microbiome, it might be a new source of antibiotics for us. And he was so extreme in this belief that the hyena had such a clean mouth

that in the context of doing this job of trying to get microbeams, he got bit by a hyena. He went to the emergency room and they're getting ready to give him antibiotics and he said, no, I don't want antibiotics. It's going to desmate my microbeam. Furthermore, I know that because I got bit by hyena, I'm not going to get an infection. They have the cleanest mouths and they had to sign all these waivers. waivers and apparently he never got an infection but the mouth microbiome is a source of potentially bacteria but maybe other sources of news science. What is the classification? What's the org chart of these bacteria? Now we're going to get back to seventh grade biology where you have the phylum, the family, the genus, the species, the strain.

And so that's the organization of them. And so I don't even remember that's the organization of them. And so... I don't even remember that anymore. Say it again. It's the phylum, the family, genus, species, strain. And so what DNA sequencing has enabled us to do is to really look at strains. And maybe one interesting thing is even within strain identities, so we give names to these strains based on their genomic makeup, even that part is really evolving in our understanding as well. So for example, you know, methylation sort of a hot topic when it comes to humans.

Bacterial genomes are also methylated. It's typically used for silencing certain genes. But you know, you sort of have this question of, is it really just the genetic makeup or these post-modifications also changing a strain from one thing to another? Should you define a strain by its genomic makeup, which is the traditional way we define things, or should you define it based on what it's doing and its actions, and that becomes more complicated. So, I think we're going to see that definition maybe evolve as we learn more. Do you mostly operate in a world where you're thinking of the strain then?

Yeah, we operate in a world where we're thinking about the strain and the function. So we, for example, as we manufacture our strains, we do occasionally these sort of whole genome audits because these strains do divide and replicate most of them basically every two hours. You want to make sure that whatever genetic modifications are just tapping naturally, don't actually impact the function of the strain. So we literally do these audits, we'll do a whole genome sequencing of these batches, we'll do a full panel of biochemical assays to understand, are they still generating small molecules at the rate that we'd expect,

growth curves? And so I think that's one of the challenges to actually being in this area is that these guys evolve really quickly. Yeah, I mean, it's hard for me to kind of wrap my head around that because even though I acknowledge what you're saying with respect to the speed of their multiplication or replication cycle, when I think of my former life when I was in a hospital, you know, everybody knows what Merse is. But does it mean that there's like a new Merse, every pick your favorite period of time where even something every month, there's a new MRSA in the hospital kind of thing that fortunately always still seems to be

responsive to vankega-mysen, but at some point it won't be like, I guess there's VRE, right? Bank resistant, Enteracoccus. So it's hard for me to wrap my head around the speed with which these things are mutating so that within the span of a year of your life, does that mean that your gut biome is changing, not just as a result of you changing, but because they're evolving, I still don't understand exactly who's optimizing for what?

Let me reframe my question. What are they optimizing for in their rapid evolution? They're optimizing for their environment. And so again, if you take diet as the primary thing that can modify your microbiome, it's the primary thing that can modify your microbiome because that's their food. They're living in your gut. They're waiting for you to feed them. The reason that MRSA and VRE are evolving is to escape the antibiotic, which is against our best interest. But what you're saying is, at least if I'm hearing you correctly,

it could be that the evolution of our gut biome isn't our best interest because in theory, it's evolving to its environment, which is us. Exactly. It's evolving to survive what we're feeding them and the environment that we're creating for them. Interesting. Okay. So very, very fluid. It is. It is. You can go on a trip to another country and eat the food that you're creating for them. Interesting. Okay. So very, very fluid. It is. It is. You can go on a trip to another country and eat the food there for a week and come back and your microbiome looks totally different in just a short period of time. I mean, many of us have experienced that. Like you go travel, you get GI distress from foods because you're not used to having

them and you can whole hog change your microbiome. That's actually what really drew me to this field. Because when you're talking about human genomics and human systems, the way in which you can impact them is limited because it's an already existing system that's relatively immutable without some real serious external force. You have to introduce a chemical, small molecule

to change a pathway that has five backups to it in your system. But the microbiome is incredibly mutable, and we're doing it all the time. And so when you think about the ability to change it and to have real health implications, that's where it's at. That's why it's an exciting field.

Got it, makes sense. Let's talk about how one measures these things, both in the lab, so what you would do, but also maybe for lack of a better word over the counter, and what I could do. So if we were interested in understanding your genes, we could do them in a number of ways. We could do the gold standard, which was we could do a whole genome sequence. We could sequence every nucleotide of every single coding and non-coding

gene in your body. And 20 years ago, that would have cost close to a billion dollars. Today, that's about a thousand dollars. Still not the most practical test in the world because it yields a whole bunch of information. One doesn't know what to do with. There's the gold standard. Conversely, we could go on a fishing expedition and you could say, well, I'm really worried about my risk of cancer and we could do a commercial test that looks at a whole bunch of known polymorphism snips and we could screen for 100 of those things

to a much more targeted look and we could get that information. Walk me through the menu of options that you as a scientist would embark on to do this and then what a consumer can do. It's actually quite similar when it comes to understanding the microbiome. So you can do shotgun sequencing where you're getting the entire genomes of all the different microbes and then you need to be able to assign which genome goes to it. Microbe, and there's a fair amount of redundancy, and so you use long read, paired with short grade sequencing to be able to get. Tell folks what that is, so they understand, yeah, because it's a bit complicated.

It's kind of exactly how it sounds. Short read DNA sequencing gives you, so let's assume, let's take an example. You've got a thousand bases of DNA that you want to sequence. A bacteria has typically how many genes and how many base pairs is. Really is a huge range of sizes. If you're trying to measure a certain piece of DNA, you can use short-weigh sequencing technologies

which allow you to, with high accuracy, get these short pieces of your sequence. So you get maybe 200 base pairs. And so you would have a few of these and then you paste them together based on kind of your sequence. So you get maybe 200 base pairs. And so you would have a few of these and then you paste them together based on kind of the overlap. And each piece is pretty accurate in terms of the sequence. Then you can do long read sequencing, which will allow you to get in one shot that entire thousand base pairs, but that tends to be a slightly lower fidelity read. And so you might have some errors in there. And so that's why kind of

the best way to do it is to do both. And I would say the reason that stitching together is problematic is because bacteria have redundant genes. And so some bacteria might have five copies of a gene versus another bacteria that has ten copies of a gene, and turns out it matters. And so, not going to remember the name of this bacterial strain, but there is a strain that's been studied pretty well that metabolizes dejoccin. A drug that's used to treat arrhythmias. Yes, and one of the reasons why that drug fell from being first-line is because the efficacy sort of had such a wide broad range across people,

so it was hard to know what's the right amount to prescribe to somebody. Wait, does it treat arrhythmias or heart failure? It's heart failure, yeah. That's how long I've been away from the game. What they found was that there's a microbe that can metabolize de-jocsin. And so people who needed higher doses to have efficacy had higher amounts of this strain. But then in another double click of that, it wasn't just that strain.

It was how many copies of this particular gene that it had. So that it over five, it can metabolize de-jocsin, less than five, it couldn't. And that means that that bacteria exists high in the GI tract, presumably, because if that were just something in your seacum or beyond, presumably, it wouldn't have impacted it. I mean, it has to be somewhat in proximity to the liver. It has such an impact on the bioavailability of that drug, right? Isn't a juxtapin an orally? Yes. I actually don't remember the location of this spectre, but I remember this part about the

number of genes being important as to whether it could even metabolize a thing or not. Because normally we think a lot about the different genes in the P450 system, in the liver, which is heavily responsible for how most drugs are metabolized, and that clearly explains a lot of the variability in human metabolism of drugs. I never knew about this. This is even one step beyond that, and obviously not dependent on our genome, but the genome of this host. Yeah, and these are mostly gut microbes,

so it really is, especially for these oral drugs, there's some path that it takes where it's interacting with these microbes, and they are able to metabolize these various drugs. Getting back to your sequencing question, the number of replicates matter. So if you imagine you have these short snippets, it's sort of a guessing game as to,

well, gee, is this an actual just replicate or was it only there one time? It was the second one, yeah. Exactly. And so that's why you need the long sequence. It becomes your template to put these short reads on. That costs several thousand dollars per sample to do. Even today.

Even today. Even today because you're really trying to get a comprehensive. And then just imagine that. And now it's every bacterial strain out there. And PS, there's a lot of redundancy and bacterial strains for a large number of them. We don't even know much like the human genome. We don't even know what are these real bacteria. What do they do? And so there's just still a lot of uncatalog genes. And so that's a rule endeavor. The second way is to really look at a QPCR-based, where you're really looking at a specific strain and trying to understand how much of that exists. Tell folks why the PCR would work that way, like what you have to use and how you have primers,

and why that's analogous to the, let's look at your cancer genes approach. Yeah, so you might have a lot of bacterial strains in your ecosystem, but you don't know how much they are relative to each other. So you can get a catalog of everything that's in there, but you don't really know how much of each one is in there from these sequencing methodologies. Although there are some really interesting tools that people are starting to develop to try to get at that, but the more accurate way to get at the quantitation or how much of this strain is actually in the microbiome is this quantitative PCR. It basically makes primers that are specific to that strain and then you're using PCR in a quantitative

way to understand, well, how much of it is in there compared to say some other strain or compared to the entirety of all the different bacteria in there. And now you have an idea of, is this constituting 1% of my microbiome, 10% of my microbiome? And so you get this quantitative piece. If you're low in specific microbes or specific functions, this becomes the way that you would really look at that. But there's an added, very important complexity that the microbiome has that the human genome doesn't have. And that's around this replication and ever changing thing. But there's an added very important complexity that the microbiome has that the human genome

doesn't have. And that's around this replication and ever changing thing. And so it's really not good enough as we talked about with the human microbiome project. It's not good enough to get one snapshot in time. You really want to understand what is your baseline microbiome. And then if you ever change your diet, you travel, you go on antibiotics, there's all kinds of things that can change your microbiome in very cute ways. Obviously, it becomes a different microbiome.

So, this longitudinal data really matters. And then, additionally, because every microbe is replicating at a different rate, the constitution or the fraction in which one microbe might be in your gut today might be different tomorrow. And so, you need longitudinal data that gets you the quantitative piece plus who are the players. And then, the third part is because they're also mutating, you need to understand what did the functions change. So if you really want to understand the microbiome, I mean, we were spending five to six thousand dollars per sample to understand

longitudinally what's happening, what are the different functions, how are these things changing? So it's pretty hard to do. And what are the ideal functional assays? Because I think it's important that we don't lose sight of what actually matters. It's what clearly matters when we look at our cells. I think we're really understanding that today in humans that function matters more than genome. The protein is more important than the gene. What do we look at in the gut biome?

Are you looking at secretory products? How are you determining the health of the function versus just the genome, which obviously must be correlated with it, but not exact, right? We are super interested in carbohydrate metabolism. And so we look at the output of that as the short-chain fatty acids. So butyrate, propionate acetate. And so you can take an individual strain, feed it as substrate, depending on where in that biochemical pathway it is, its substrate might be slightly different, and then look at how much of those small molecules are being produced on a gas chromatographer. And so you're running a enzymatic reaction, if you think about the bacteria as the enzyme, you're running a enzymatic reaction. If you think about the bacteria as the enzyme,

you're giving a substrate and you're basically doing your old-fashioned macalusmen and curves where you're giving it increasing amounts of substrate. And you're basically looking at the enzymology of going from substrate to that short-chain fatty acid. So let's use that as an example. You eat a piece of bread, starch, polysaccharide, digestion, of course, begins in the mouth. AMOLED starts to break that down.

It continues further in the stomach. By the time it gets to the jajunum, I mean, it's mostly just glucose monomers, right? I think that's still being figured out, actually, what is the state of affairs and things arrive? I mean, it's actually really hard to survey what's happening in the gut microbiome because we don't have good sampling methodologies. That's super interesting. So you're saying, even if you dropped a tube down

somebody's throat and you just sampled the slurry at the distal end of the duodenum or the proximal end of the jajunum, I mean, I've seen what it looks like when you operate on somebody. We can't tell what the composition of matter is there. I'm not squarely in this, but I haven't seen anything pop out that was really compelling. The pushback all of those technologies get is the ease with which things can get contaminated.

And so essentially, you might be able to get a sample, but then in the process of pulling that back out, it becomes contaminated with the other things that are along the track. Could be done interoperatively. Hopefully someone is doing that, right? If you're in there operating otherwise. Now now the problem is you wouldn't be operating on somebody without a bowel prep. And so you've completely destroyed the system. Also, you're not operating on somebody in an anaerobic chamber.

The second they get exposed to oxygen, it's sort of different. Okay. So explain how carbohydrate metabolism produces byproducts and what those byproducts are, because people who listen to this podcast, people like me, when I think of carbohydrate metabolism, I don't think of any of the things you just said. I think of glucose, I think of glucose one phosphate, I think of glucose six phosphate, I think of pyruvate,

I think of acetyl CoA, I think of the crept cycle, I think of lactate as a byproduct of oxidative phosphorylation. So, you're thinking of things in a different level because you're obviously looking at a different host. So, explain the metabolism on that side of the ledger. So, in this case, when we talk about carbohydrates, we're really talking about these fibers. And so, our microbiome is uniquely positioned to metabolize fibers, a wide variety of which

we actually can't even metabolize ourselves. And fiber is one of the primary prebiotics that feeds all of your strains. And so you're right, we think about carbohydrate metabolism from the perspective of the microbiome, it's really thing about these fibers. There are primary and secondary fermenters in the microbiome that can metabolize these fibers into certain short-chain fatty acids, which then become precursors for the ultimate short-chain fatty acid, which is butarate. Butarate is this incredibly important short-chain fatty acid that's been studied in a

wide variety of conditions. And so your microbiome, one of the most important molecules it makes, is butarate. And so butarrate has a role in GI health. The colon cells are the only cells that use butyrate as their source of energy as opposed to glucose, which is what you use by every other cell. And so when you don't have enough butyrate that's been associated with things like colon cancer and not having good colon health.

But butyrate is also a small molecule that triggers G-protein coupled receptors to then release GLP1 from these L cells in the microbiome. So it also plays a role in sort of this gut metabolism axis. Beterate becomes this really important small molecule that the gut is producing based on the foods that you're eating, and that's kind of where we've really honed in. So if we think about the difference between soluble and insoluble fiber, are you referring

mostly here to insoluble fiber because you alluded to the fact that it's fiber we aren't able to digest and metabolize. That is the food stock for these bacteria. Yeah, that's really where we're focused on as these insoluble fibers. Insoluble fiber, I guess, is most readily available in vegetables, correct?

Exactly. Vegetables and fruits, yeah. I was asked recently by a patient, I drink this green drink every morning and they're like, why do you drink it? Do you think it's a substitute for having vegetables? And I said, I really don't. I said, I think it's a substitute for having vegetables with respect to the vitamins you get and probably even a lot of the polyphenols. but it's clearly not a substitute for the fiber, just based on the practicality. You can look at the ingredient label. There isn't enough fiber in this to be a substitute. So at the end of the day, there doesn't appear to be, if you buy

the argument, which I think we're going to discuss, that fiber is essential for gut health, which by extension means essential for human health, you have to be eating a ton of fiber, insoluble fiber. Yeah, you do. Although I mean there have been a Meredith studies showing that getting fiber from these external sources, these powders, then you have to get them in the volumes needed. They can be pretty impactful for your microbiome as well. What are typical recommendations of fiber like how many grams per day, 18, 30, something like that? I actually don't know what the current standard recommended dose is, but I think it's somewhere between 20 and 30 grams. Yeah, it's hard to imagine getting that in a supplement.

And I think actually your average American's consuming like one to two were way off base on how much fiber we're consuming. But just to get your question, though, there's also a delivery component to this too. So taking the fiber where you mix it in a drink might be a lot less impactful, or you might need a lot more of it, I should say, then if you were to take it in a enteric coated capsule that got through the stomach acid and then closer to the distal colon, it's a delivery question. So if you might be able to get away with a lot less. Oh, interesting.

Why is that? I mean, I thought that given that it's insoluble fiber, it would be impervious to the gut and the environment all along the way. Is that not the case? The measurements of these things along the way has been elusive. That's my daughter would say, sus.

It's sus. We include Inulin in our pill, an incredibly small amount. It's in the order of one to two hundred milligrams, so it's definitely not at the dietary fiber level. But the reason we included is because when we did our preclinical studies, we saw that if we didn't include it, the strains really didn't colonize as well. So the concept behind that is that by having the Inulin right there next to the strains, as soon as that capsule dissolves, these things start to get hydrant and start to become alive, and very close proximity is their food. And that's why I think about that kind of delivery and that proximity question

as being something we haven't really solved for yet. Interesting. Okay, so you eat a piece of celery, which is basically all insoluble fiber. You mechanically break it into smaller pieces, but functionally, it's still cellulose that's making its way down. At what point in the gut, does it have to wait till it's in the colon, or does the metabolism and production of butyrate begin in the small intestine?

There's some bifidobacterium in the small intestine that can help start to break that down, but I think the general thought is that the majority of that metabolism is happening in the distal colon. Okay. And which strains of bacteria are most responsible for the metabolism of insoluble fiber in the production of butyrate as a byproduct? It's a multi-step pathway. So there are clostridial strains which can do this breakdown. There are bifidobacterium strains which can do this breakdown. There are bifidobacterium strains that can do this breakdown. And then there is the strain acrimancia,

which was discovered in the early 2000s. And it also plays a really important role in generating these short-chain fatty acids. And are these strains that they belong to the same species? These are the species names. Okay. Strain name would have two parts to it. So if I said acromansium, use an aphilla, that would be the name of the strain, but I've shortened it to acromansia, which really does represent the species, the higher level. Got it. So E. coli is... Strain name.

Is actually a strain name. Yeah. Ashera, whatever it would be, the species. Got it. Clostridium is the species. Clostridium difficile is the strain. Dr. Justin Marchegiani Got it. Clestidium is the species. Clestidium difficile is the strength. Dr. Justin Yep. Okay.

Let's pivot for a moment and actually talk about C-Diff because that's one of the most compelling arguments for an intervention in treating human disease by manipulating the gut bacteria. So tell folks what Clestridium difficile is as a bacteria and you already alluded to this, but how does it go from being a benign slash benevolent participant in our coexistence with the universe to one that could kill us?

So clostridium difficile is a strain that exists and many of us have it in low levels in our gut microbiome and we walk around perfectly healthy and fine with it. When you take an antibiotic, that's essentially almost like a nuclear bomb to your microbiome. It kills everything in there, but in some cases it doesn't kill everything. And so there are the strains of clostridium difficile that after you take an antibiotic, it's killed all the different strains off, but it didn't kill your clostridium difficile strain. And the problem with killing off all the other strains is now all the competition is gone.

So you imagine you have this ecosystem of different microbes, and now you just got rid of all of them. So now you have this strain that has no competition, and it can start to propagate unchecked. And so it's when you start to have these really high levels of this strain, Clostridium difficile in your microbiome, that's when it starts to make you really sick and actually ultimately it's fatal. And so the way in which we treat infections is through more antibiotics. And so when you have this clostridium-difficile infection, which is a result of having taken an antibiotic almost ironically, the cure also is another antibiotic.

And really what you're trying to do is to get an antibiotic that can kill that strain and get it back to its low levels until your microbiome can reconstitute itself through the food that you eat. The success rate of those antibiotics varies from person to person, but overall it's something like 70% successful. And remember fatality is on the other side of this. I was just about to say, make sure people understand what non-success means. Yes. Yes. 70% success is not great when the 30% means that you're gonna die on the other side.

So one of the concepts was really to go in the reverse direction and say, okay, if the problem is that now there's no competition and this guy can propagate unchecked, what if we just load up the person's gut with all these different microbes, reestablish an ecosystem, and then that's the way to kind of temper this thing down.

And that's called a fecal microbiome transplant. It's exactly what it sounds like. You literally take feces from a person and transplant it into another person. That thing has like a 99% success rate. When was this first done? Phil, this is at least in the 90s, maybe earlier? Well, it was done as one-offs before people were doing real studies with it. And actually a ton of them were done in Australia. It's like one of the places where the most

of these are done. But it has an incredible success rate. It's kind of gross. Yeah, what was it like back in the day and how has it done today? I assume today it's done through capsules. Well, I think people are still using the methodology of actually going up through the colon, but people have tried to create capsules and try to be able to make it so that you can just consume these capsules with a drink and then it gets there. And those seem to be effective. They're not as effective as the enema version.

And because probably in the process of getting that stool into a freeze-dried format or into a pill format, you're losing probably some of the diversity that you just get when you do it the old fashioned way. And there are risks associated with this or not? Are they overblown? I mean, there's certainly been hoopla about this at the FDA at one point. What was that about? Fundamentally, it doesn't feel like a therapeutic to give people shit. I think there's this instinct that, okay, this isn't safe, but there's a reality around it,

which is that the source of that person is really important, right? How do you know that that's a healthy, so-called healthy stool donor? And so, what if they've got some weird pathogen that now you put it into this person who's already into pleased state? He's already compromised, yeah. And then furthermore, because everybody's microbiomes do have these different functions,

it's possible that you might cure them of the CDIF, okay, great, and you might not have given them a new pathogen,'s fine, but you might have changed their metabolism of foods in a way that's not beneficial to them and so For example, there are some of these case studies of someone getting a fecal migraum transplant and now all of a sudden They have obesity issues that they never had before But really is at the heart of it is that we're very early in understanding microbiome science and so I think the FDA You know their job is to make sure that people are safe. And so they have to decide, is it more safe to give people this cure that has this incredibly high success rate when fatalities on the other side?

Or is it more safe to say, hey, let's keep learning more about this before we actually introduce it into clinical care? Have they actually tried to shut the whole thing down? They essentially said, FMTs are not approved by the FDA. They're not safe until we learn more about the science. What year was that? Ish.

This was probably, I don't know, maybe 20 years ago. Essentially, these people who had these physicians, these patients, patients' families who had all experienced this benefit. And then people who were in the queue to get it done, essentially with pitch forks in hand went to DC and said, this is crazy, like this thing has such a high curate, how could you possibly take it away as an option for people? And so they put their tail between their legs and reverse it and let people do them.

Let's talk about an example you just gave there, which I know was sort of theoretical, but I'm sure there are many cases like it where you take a lean individual who's hospitalized for some reason. They're given an antibiotic in the process. They develop C. Diff colitis. They get a fecal transplant and they recover. The fecal transplant came from somebody who was obese and now they develop obesity. So the hypothesis here has to be pretty clear that the gut bacteria of the obese individual is playing a causal role in their obesity. So I wanna tackle that topic, but first, why wouldn't the habits of the recipient immediately override the bacteria

that they got on the receiving end given that their habits are more in line with being lean. In other words, given that rapid evolution of these bacteria, why do they persist in their phenotype? Well, first of all, these are case studies. These are not clinical trials, and so I think you're dealing with the uncontrollable nature of humans. But I would say one of the interesting things is that these people's habits did not go back to what they used to have. And one of the things we're learning about the microbiome is through this gut brain connection,

your microbiome can actually influence your food cravings, and we actually have some data supporting that as well. And so if you think about this new microbiome as maybe it's not metabolizing food as efficiently, it gives you predilection towards obesity. But on top of that, it changes your food cravings. You've actually now got a double whammy against you that keeps you from being your old self. And so I think the microbiome, this gut brain connection,

it's ability to generate neurotransmitters, it's ability to change cravings. It's still at early stages of understanding, but it can basically change your behavior, so you're not really the old person you are. Profound. Two thoughts. The first is, could you be in that situation forced to eat your old way?

You're going to go to a one-week boot camp where you're literally force-fed. Everything you use to eat, would that be sufficient to return your gut biome to where it was? Talk about how the gut and the brain connect. So my vague recollection is serotonin plays a role here. You've already talked about GLP1, GIP typically is secreted higher in the gut, but that presumably plays a role as well. Again, this is in the early stages, and we're going to learn so much more over time, but the gut is a massive producer of a bunch of things that we've traditionally thought of as neurotransmitters.

So yeah, serotonin, dopamine, GABA, these GLP1GIPs. And then I would also say there are neurons in the gut. And so there's this relationship between the neurons in the gut and the neurons in the brain. The neurotransmitters being generated by the gut can make their way straight to the brain through the vagus nerve. And so there is potentially an opportunity, for example, to tackle stress and anxiety through the gut microbiome where you have these gut microbes that can produce large amounts of GABA, which has been shown to be able to reduce stress and anxiety.

And so is there an opportunity to go after the gut to reduce stress and anxiety? And again, kind of getting back to this cravings thing, it's really interesting for anybody who's ever felt stress or anxiety, you know, actually your food cravings oftentimes change and kind of a hand in hand with that. So this part about the gut brain connection that involves both cravings and these neurotransmitters I think is really, really fascinating. How much of that do you think is regulated by things outside of food? Meaning, the gut flora is it comes to changing function, which then changes our physiologic

state. I mean, we've talked about food. We should talk about it some more clearly, a very important piece. You've already talked about antibiotics. What are other modifiable factors that change this? Certainly, the two biggest changes in microbiome are the things we've talked about, antibiotics and nutrition. But beyond that, what we know is that there are,

and this is especially with regards to diversity and loss of function of your microbiome. We know that as you age, your microbiome starts to become depleted. We know that when you go through periods of intense stress, your microbiome becomes depleted. We know that when your circadian rhythm gets disrupted, so you travel and day becomes night, night becomes day, that changes your microbiome.

By the way, when you say depleted, tell me what that means in a technical sense. It means that what happens is that you have loss of function. So you start to become depleted in specific strains that have specific functions. And so, for example, this strain that I alluded to earlier, acrimansia is one of the ones that is deeply studied. And low levels of acrimansia have been associated with a really

wide variety of diseases ranging from obesity to immunological disorders to stress and anxiety and GI issues. And so basically the depletion means you're losing certain strains and certain functions. And then for us women, when we go through menopause, there's like a massive change in the microbiome on the other side of that lovely experience. I was going to ask you about the vaginal microbiome, but right now, are you saying that with respect to gut as well? I'm saying that with respect to gut, yep. Okay, what are the factors that influence the vaginal microbiome besides the most obvious which must be menopause

pregnancy or anything that has a dramatic shift in endometrial lining and changes? So presumably if you sample a woman's vaginal microbiome across the month of her cycle given that her estrogen progesterone levels are fluctuating you're gonna see differences but I assume there's a quote-unquote healthy versus an unhealthy. How does that manifest itself? And what does one do about that? Well, I'm not an expert in the vaginal microbiome, but certainly these lactobacillus strains are

some of the more important strains in the vaginal microbiome. And the name of the game there is actually production of acid. You want an acidic environment there. So it's a protection environment. It's a protection environment. It's the goal is to keep yeast in particular and other things away. Exactly. Spirm everything.

So there is a healthy vaginal microbiome, which is really associated with the acidity level that these bacteria keep the vaginal microbiome at. It can be influenced actually also by the food that you eat. I don't know. It could be that I don't know, but it's not totally elucidated. But for example, you might know that one of the things that gets recommended for you today is cranberry juice, and you're not inserting that into your vaginal canal. You're consuming it.

And so there's definitely this connection between the food that you're eating, and then the composition of your vaginal microbiome pregnancy is also another thing that really changes it. And one of the implications that we did look into, this is some work that we were doing with the Mayo Clinic early on in our company, is the idea of preterm labor. So preterm labor has been highly associated with bacterial vaginosis, and that is essentially the broad name because I think we don't fully understand what it means, but it's an infection of the, or a disruption of the vaginal microbiome, this infection triggers preterm labor.

And so one of the ideas here is that, well, if you could prevent bacterial vaginosis, you might be able to reduce the incidence of preterm labor. And there's other idea, which is that once you have a preemie, you're a lot more prone to have a preemie again. And these women usually also have this bacterial vaginosis, which happens right before they deliver the baby. So maybe there's something about your vaginal microbiome that makes you more susceptible to bacterial vaginosis, which then leads to this preterm labor.

So then there's, of course, the UTIs and yeast infections and things like that. Any other systemic complications from a negatively altered vaginal microbiome? I mean, I guess you could argue preterm laborers systemic. Outside of the pregnancy case, if you just talk about a woman who's entered menopause, because I'm guessing being on hormones versus not on hormones

is going to have a profound impact on that. Do we know if any of the problems associated with menopause beyond the obvious ones, like vaginal dryness and hot flashes, have their roots in a change in that microbiome? I think that work is still pretty early on. So I would hesitate to go too far there. And even the gut microbiome too, there are microbes that can, I'm not an expert in menopause

or these hormones, but I believe that estrogen has a modification that gets made that is a signal to the body to not put that estrogen back in a circulation, but to, I guess you must urinate it out. But there are microbes in the gut, their job is to remove that modification, and so the effect is that it could potentially increase your circulating estrogen. And so this is all super early stage. If you go try to look up studies around menopause in the microbiome, it's extremely sparse. And so I think we just really don't know. But there could be a day, presumably, when there are bacterial products that are used to

address some of these issues, it certainly would make sense. Going back now to the example, you gave how robust is the evidence in humans, and maybe we can start by the evidence in mice, because it gets all the headlines, where you can take an obese mouse and you can do a fecal transplant from a lean mouse into an obese mouse and make that obese mouse a lean mouse. Is that a relatively robust, reproducible finding? Yeah, a lot of people have done that even to take it one step further, they can make these mice obese or lean using a human microbiome transplant. So they basically would take the microbiome from an obese woman, put it into a lean

mouse, and see that lean mouse become obese. And so we talked about the overlap between mice and men. There is some overlap there because you can actually change the metabolism of these mice through these fecal transplants. Why has that not taken off in humans? I mean, what are the impediments to that? Are those studies being done? People have done fecal microbiome transplants into people with type 2 diabetes, for example. I think there's two big challenges. One is, of

course, the safety challenge, which is, do you really feel confident that that stool donor is a stool that you want to have in your body and move forward with. But more profoundly, and maybe linked to that, is the ability to reproduce the effect. So even if it's always Joey who's providing the stool, I mean, Joey travels, Joey goes on antibiotics, Joey changes his diet, you may not always be eating the same stool from that donor. And so in order to generate consistent results, we do in drug development, dose response curves. You're really trying to understand in this small molecule in this setting, how much of it do you need and what effect will it have?

You can't do that if your intervention is a fecal microbiome transplant because that thing is different from person to person and even from the same person. It's a pretty messy drug. It's a messy drug. When we started this company, the idea was the FICO microbiome transplant is super compelling data. That tells you that there's something in that microbiome that can change your physiology. But the name of the game is, how do you figure out what are the components of that microbiome

that are conferring that benefit and how do you make a reproducible manufacturing and quality process so that you know every time I deliver this pill to you, I know exactly what's in it. And it's the same every single time. And so that becomes a challenge. How do you take that whole kitchen sink, the whole micro, fecal microbiome and say, actually, it's these four strains or these two strains or these 50 strains. That's the tricky part.

So let's talk about what those strains might be and what their function is. So for folks listening, what are the strain names that we have to know here? part. Let's talk about what those strains might be and what their function is. For folks listening, what are the strain names that we have to know here? Lactobacillus, you've already talked about. Acrimancy, you've already talked about. Yeah, I think if you start to look at labels on the supplements that are out there, you're going to see a lot of lactobacillus and bifidobacterium. Those are the two primary species that are pretty much in every product.

Those strains are relatively easy to grow. It's one of the reasons why. Those are the strains that are in a lot of. Those strains are relatively easy to grow. It's one of the reasons why those are the strains that are in a lot of probably why are they easy to grow. I thought anaerobes are hard to grow. These fall into that in between their facultative. You can grow them in some amount of oxygen. In fact, an acrimansia is not. It's a strict anerobe. So in fact, when we first started wanting to manufacture acrimansia, we just tried to outsource it to all these probiotics manufacturers around the world and they kept sending us back dead stuff. And it's because in order to manufacture a strict anaerobic, your entire manufacturing system end and has to keep all oxygen out.

A single molecule of oxygen kills the whole batch. And so there's a real manufacturing challenge to some of these next generation probiotic strains. Before we talk about that, let's talk more about the Bifro and lacto. Are they a bit of the shiny light problem? The keys under the shiny light. Yeah. Yeah.

So, are they basically the ones that are most ubiquitous because they're easy to grow, or is there a really good physiologic case for their utility? They're easy to grow, and so they're prevalent. That's why so much work has been done on them. Because there was no microbiome science when these strains started making their way into the market in the 70s. There was just, what can I culture? And that's what's culture-able. Nobody thought about strict anerobs or how to keep oxygen out. That's a pain and that's expensive. So these are things that have been around since the 70s. And furthermore, they got grandfathered in by the FDA as safe because they've been on the

market for so long. You want to bring a new strain to market. You have to go through the entire process of the FDA that says, hey, it's a new strain, but let me demonstrate to you all the thousands of reasons why this thing is safe. Just to be clear, from a regulatory standpoint, if a company wants to put a new strain into a probiotic, they're going down a grass pathway, generally, regarded as safe pathway, or an IND. If you want to be able to sell it directly to consumers, you have to go down the grass pathway. Are there people that are actually looking at a pure pharma strategy where they're making an IND and going down

the drug pathway? Absolutely. There are pharmaceutical, actually, they're all startups, but pharmaceutical drug companies that are using the microbiome as the therapeutic and filing INDs and doing these studies. And in fact, this fecal microbiome transplant in the form of a pill, that's a therapeutic. There are multiple companies out there that are developing that as a drug therapy. Interesting.

I mean, the closest thing that I can think of to that would be immunotherapy using TIL. You have immunotherapy in the form of a drug where you use something like K-Truda, um, C-T-L-4 inhibitor, where you have a single molecule that is an actual drug that triggers the immune system to do something. But then you have tumor infiltrating lymphocytes where you go and harvest to patients, tumor, grow those T cells out, multiply them, you know, a log or more, and then re-infuse them, actually several logs, and then re-infuse them. But of course, then every patient has their own designer drug. That's still a purely regulated FDA,

IND problem. That sounds a lot more like the fecal microbiome transplant, which is every patient has a different drug. The difference there is it's not coming from them. By the way, begs the question, should every patient going to the hospital have a bank to stool sample so that they can be their own transplant? You could have an autologous transplant and save a lot of the risk. Yeah, I think that would probably be a super smart thing to do. Bank your own stool. You could do it in your own freezer. Right, just leave a stool sample in your fridge when you go to the hospital.

And if you can make it home and you're fine, you can pitch it. Yeah. Yeah, interesting. What do your interests in this other species called acrimansia? I think that acrimansia has become an incredibly interesting strain over the last decade. But when we first became interested in it, it was really centered around trying to understand the difference in people with metabolic syndrome versus people who don't have metabolic syndrome. We did in a variety of people who have done these twin studies.

These twin studies are really interesting because you take a twin pair and you're looking for discordant twins or one twin is healthy, the other twin has diabetes or a zobies. And you're looking at their microbiome and you're basically saying, hey, man, genetically, these two people are the same. This is a nurture, not nature problem. And so you start to look at their microbiomes and you start to see patterns around the world. If you compile the stated together, you start to see that the twin that has obesity,

your type two diabetes has a different microbiome that the twin that has obesity or type 2 diabetes has a different microbiome from the twin that's healthy. And one of the hallmarks that appears to be true across different cultures and different dietary patterns is this depletion in this strain called acrimansia. In absolute numbers. In absolute numbers. And what's the relative difference between these populations? That I don't know, but I do think that it is interesting because across these different

populations, acrimansia is, we don't have to get into this, but the so-called healthy gut microbiome, which is yet to be defined, across the world, acrimansia comprises the microbiome on the order of somewhere between like 5% to 10% or 5% to 8% for a healthy individual, no matter where you're living. And so being depleted or having nothing show up is a really stark difference. Oh, it's that profound a difference. The unhealthy person has none of it. The healthy person has 5 to 10%.

Yes. At 5 to 10%, does that make Acrimencia the most predominant species in the gut? I think it is among the most, actually don't know that there's a current telling of winners because there's such a difference from person to person in terms of the composition of the microbiome. Now, you said something. It's second ago. It's very interesting, which, as you said, regardless of where a person lived, if they're

a very metabolically healthy individual, I'm paraphrasing a little bit, they're about 5 to 10% acrimansia. But presumably that also means it's independent of diet because you can point to people who are on radically different diets who are metabolically healthy. So how is that explained? You can have people who are on an all-plant diet who are metabolically healthy and presumably eating who are metabolically healthy and presumably eating a hundred grams or more of fiber a day, plenty of food. And you can have somebody on a carnivore diet, and I've seen people on these who are very metabolically healthy. I think it's an impossible diet to adhere to for someone like me who are not eating a gram of fiber, but by all measurements are metabolically healthy. And then of course everything in between. So given the sensitivity of this flora to diet, how do we make that explanation?

I trying to think about any studies that really got at the heart of this, really the study that you would want to do is you want to take one of these individuals, measure their microbiome and you'd want to put them on a series of different diets and just see, like, is this something that is not changeable? Like, no matter what you eat or what diet you're on, this particular strain stays high. In which case, you would argue that the microbiome is a readout of their metabolic health, not the cause of their metabolic health. Could be, I was going to go in a slightly different direction, which is to say that

perhaps the host themselves play a much larger role in the composition of your microbiome than we really understand. So as you change your diet, you would expect your microbiome to change. But if there's something about the host, that part never changes, that might be the influence, to be honest, I just don't think we know. Well, I mean, that's interesting. And by the way, it's possible that these studies haven't been done across a broad enough dietary spectrum.

So even though there's geographic diversity, maybe there isn't as much dietary diversity. And that would be interesting as well, I suppose. For example, not that we could go back and do this, but if you go and look at the Inuit who, prior to the adoption of Western food, we're eating a seemingly ridiculous diet, and yet we're quite metabolically healthy. Yeah, it does make me wonder if I could go back and look at this too. I mean, there are tribes in South America that have been relatively untainted by anything

outside of where they are. And so I know that there are definitely microbiome studies that are happening in some of those tribal groups. That could be sort of the beginnings of this kind of a study. Quickly the difference between a probiotic and a prebiotic. Oh boy. Yeah, the vocabulary lesson. I'll introduce another one that's become popularized, which is the postbiotic.

So the microbiome, when we talk about it, is all these bacterial and yeast strains. That's the probiotic. So the probiotic is the living organism itself. Prebiotic is the food that feeds those organisms. So we talked about fibers and inulin and polyphenols and things like that. Those are prebiotic. So prebiotics are the food. The probiotic is the organism itself.

And then what these organisms produce or what they secrete is now being called the postbiotic. And so you would call... Buturrate. Buturrate would be a postbiotic. Yeah. And then maybe one more term people might start seeing is sinbiotic. And all sinbiotic refers to is you've mixed two or more of these things together. The pre and the probiotic together are the probiotic and the postbiotic.

So a sinbiotic has multiple. When people are consuming yogurt, they're consuming a prebiotic presumably. It's the bacteria in it that they're trying to- Yeah. They're lactobacillus bacteria that are in that yogurt that stay alive in the context of yogurt. And so that's really what you're consuming. And what is the perceived, believed or realized efficacy of consuming massive amounts of lactobacillus and diphtobacteri factor? I think the most well documented and even reported from consumers impact of consuming lactobacillus and diphtobacterium,ics in general, and yogurts is around GI symptoms.

So things like gas and bloating and diarrhea and constipation. A lot of people report, and there have been studies showing both sides of it. But basically, there have definitely been studies showing and people reporting. They have better GI when they're consuming these probiotics or these yogurts of probiotics. If you go and buy buy yogurt off the shelf, how much lactobacillus is naturally within that? I think they added in, so it varies. It's not something that naturally occurs in yogurt. I think it is naturally occurring in yogurt, but all the things you're buying off the shelf, they're also supplementing with additional lactobacillus.

The real question to ask yourself when you're buying yogurt is how much sugar is in it. Yeah, for sure, if you're buying fruity flavored yogurts, by the way, I want to come back and talk about sugars in a second, but I want to go back to kind of this probiotic. How much? What is the dose effect, right? So I know that if you look at a bottle of pick your favorite probiotic, it usually uses something called CFU's colony forming units. Can you explain what those are?

Yes. Some brilliant marketer decided that that was going to become the metric, the name of the game for probiotics. So colony forming units remember back to seventh grade biology where you might have been given a peachy dish and you had to like swab your mouth or swab your hand or put your finger on it and then you see what grows. That's basically that tool. So you take your pill or whatever your yogurt and you basically spread it out on a peachy dish and then you count how many colonies form and that gives you a number. So you'll say, well gee this pill has 10 to the ninth colony forming units in it. So some marker decided that that's the most

important thing is the number of colony forming units and I have 10 times more colony forming units than somebody else. Maybe that's interesting, but it's less relevant than the function of what's happening in your pill. And moreover, it only gives you one piece of data about what's in that pill. So when you do that, the only thing you get to see are what's able to form a colony. But actually, in almost every supplement, every probiotic out there, the majority of what's in that pill is dead probiotic.

And you don't see any of that when you use this technology. There's a different tool that can be used called flow cytometry. And essentially what you do is you take your capsule, you put it into this flow cytometer, and what it pops out, it's read out, is live cells, dead cells, and kind of in between cells. It's based on a staining of the membrane. And so it tells you which of these cells is viable. Like the membrane is really intact, which of them have a compromised membrane

and which ones are kind of somewhere in between. And now you know exactly what's in your pill, because even if you could have the same number of live cells, it turns out that those dead cells and those in between cells, they actually have a role to play. They have a function in there. So these so-called postbiotics, that's what those guys are. And so you don't really know what's in your pill unless you're using flow cytometry versus

colin-frontal. And when you do flow cytometry, what do you stain for? Which surface receptors or molecules you're staining for? I don't know the answer to that. I think there's two that we stain for, but I actually don't remember. Okay. And it varies by bacteria on the cell. Yeah. Well, there's some of these common ones among almost all bacteria, but yeah.

Okay. So lacto, it's basically able to use oxygen and not use oxygen. So obviously, if you're going to produce it, you're going to produce it under the conditions of oxygen because it's cheaper and easier. I hear things about some of these things need to be refrigerated. Some don't. What's the specificity of that? Yeah, it's really about stabilizing something that's meant to be inside the body.

So I think that a lot of these lactobacillus and bifidobacterium strains, because we've been manufacturing them for so long, we've figured out how to get them to be stable to this process. So just to maybe take a step back and what is the process, you take these strains and you're basically growing them into these big vats. If you've ever been to a brewery or a vineyard, you see these big vats, sort of the same in these manufacturing plants. You grow these strains in a cultural media.

And then when they get to a certain density, optical density, you then harvest them. So centrifugation is one of the most common ways. You basically spin down the cells, you get rid of the media, and now you have this kind of paste of cells. How slowly do you need to spin them to prevent them from dying? down the cells, get rid of the media, and now you have this kind of paste of cells. How slowly do you need to spin them to prevent them from dying? Lactobacillus and bifidose, at least the ones that are in the market today, they're pretty

hardy. You can spin them awfully fast and be done with this whole process pretty quickly. And there's other strains that are a lot more sensitive, so you have to do this process much more slowly. And so I think that depends on the strain. But you essentially throw away the media, you take this paste of cells and then you freeze dry them. And so that gets them into a powder form.

And once they're in a powder form, they tend to be pretty stable. And then you could throw them in a pill and you're off to the races. What's the yield on that? How much loss do you have in live bacteria? Assuming you start with 100% live in the paste, when you just go through the act of freeze-drying them, what's your yield? That is where everybody dies.

I mean, basically a lot of times you're losing like 90% of yourselves just through that freeze-drying process. It's a pretty harsh process. So what people do is you try to figure out what cryoprotectants you can add that are not going to harm the bacteria on the other side of it, but that they're going to help them through this cryo phase. How does freeze drying actually work? It's both pressure and temperature. There are these instruments called lavalizers and

a lot of them use these sort of flat pans. And so you would put your paste, it's like baking. You put your paste in this flat pan. You slide into the lathalyzer, and then over a period of time, it's basically using temperature and cold, the sublimation process of removing the liquid from the cells directly into a gas. Again, we're going back to seventh grade biology. And on the other side of that, you have this powder, and that powder may be stable at room temperature. It may still need to be refrigerated, but what you're trying to figure out is what are the additives that you can incorporate that help it get through this freeze drying process and remain viable. But the things that have to stay refrigerated, even after that process, they're still relatively

unstable. So you still have to keep them in the refrigerator. For us, for example, we first started making acrimansia, it absolutely had to be refrigerated. I mean, within hours, it would die. Meaning, once you freeze dry it, you sort of shut its metabolism down completely. And then the minute it gets even a little bit warmer, it basically warms enough to the point where it's no longer cryogenically preserved. But there's no substrate for it. And it dies. Is that the actual mechanism by which it dies? That's how it dies. Exactly. Because there's no substrate for it, and so it can't handle

the heat, and so it dies. The most important part is you do want it to be able to get through this freeze drying process, and of course everybody wants shelf-stable product. But the most important thing is that when you ingest this strain, it actually makes its way to the gut microbiome, is able to reconstitute and to perform its functions, to create the proteins as opposed to secrete. to the gut microbiome is able to reconstitute and to perform its functions to create the proteins that's supposed to secrete. Secrete the small molecules that's supposed to secrete.

And so when you do this process, every time you make a mod, you're like, oh my gosh, we just improve the viability 4x. You just have to go to the other side and say, okay, now you gotta put it in a human and see if I got the same output. So it's a pretty lengthy process. By the way, do you think that that 90% loss is typical on the most common strains that

are used commercially? No, and I mean, we even grow some of those strains and you can maintain higher viability. You're never going to get 100% through, but I think it's more like on the order of between 50 and 80%. So you're definitely above the halfway mark. Okay. And then how many of those make it into the person. So in other words, once they're reheated, how many of them die along the way? A lot of it depends on the encapsulation that you're

using. So if you have these interocoded capsules, you can make it through this time at GASID, and then you can use these time-released capsules so that it takes a certain amount of time and you're making some assumptions around GI transit. And then there's kind of the least expensive version, which probably all falls apart in the stomach and very little of it gets to the actual destination. So it wouldn't make sense to be drinking a probiotic and a liquid then. You could get a lot more efficacy if you took it in a pill format. Part of it too is we talked about these lactobacillus that are in yogurts. Part of it is that if you just have to get just on the other side

of the stomach and the small intestine, that is a different thing than trying to get all the way to the gut microbiome. And is that where it seems that lacto and bifero need to be seeded is just outside the stomach? I don't know that the answer to that is known, but you can actually find them all along the track and even into the distal colon. And what is their functional output? Do they make butyrate? Lactobacillus tend to produce lactate besides that, yeah. Bifidobacterium, various of those strains can produce short-chain fatty acids.

They need to be paired with secondary fermenters in order to get to the final butorate. Now, lactate is also a great substrate fuel for enterocytes, correct? Yes. I think maybe that's why these exist all along the track. In that sense, is the primary function of the lactobacillus to make food for the gut. I don't think we know exactly the answer to that, but definitely the products of those lactobacillus strains, there are a bunch of other strains that are dependent

on having lactobacillus. Does it also in that 5% to 10% prevalence? I actually don't know the answer to that. Okay. Lots of companies out there are selling lactobacillus probiotics. You should put them in the fridge and you should hope that their manufacturing process is such that you're getting 80% instead of 10% of what they claim.

Yeah, this is where I think our government could maybe have an important role, which is to put some guidelines and requirements around labeling. They'll give you an even more crazy thing. Even the CFU, let's say you think CFU is a perfectly fine metric. The CFU that gets put on that label could be the CFU at the time of manufacturer or could be the CFU, if I say this thing's got a two-year shelf-life stability, it could be the CFU at the two-year time mark. And you're really depending on, is this company going to tell me the higher number or are they going to try to legitimately tell me what the thing is at the end of the shelf life?

And is CFU by definition always alive because presumably if they form a colony, they were alive? Yeah, yeah. Are there any recommendations one would make about how many CFU you need of each of those two bacteria? Well, I think people tend to kind of converge around this like, oh, you need billions.

I don't know. I mean, I guess maybe I would ask the question of study different way, which is, what are you trying to do? What are probably trying to solve? What are probably trying to solve? Exactly. So if you've got a problem you're trying to solve,

I'm generally think most people should operate under, I just need the minimum viable product to solve my problem. So for you, that might be 10 billion for somebody else, it might be 10 to the 7th. Unfortunately, I think because of marketing, a lot of people are taking things, and they don't even really know why they're taking them. Well, it gets difficult when you don't know what the problem is that you're trying to fix. And I think that therein lies a big challenge, which is what am I objectively fixing? Now, it could be something symptomatic, as you said,

it could be, boy, if I take a probiotic, my GI symptoms vanish, I'm not bloated. Great. So just titrate to the point where that's true. So, going back to antibiotics for a second, for most people taking antibiotics is almost always oral. But of course, if you're in the hospital, it's not uncommon to take an intravenous antibiotic. So, for example, if you're having surgery, it's not uncommon right before the surgeon cuts your skin, for them to administer an intravenous antibiotic, usually something that covers for skin flora, and that reduces the risk of surgical wound infection.

Do intravenous antibiotics also have the same obliterating effect on the gut biome that oral ones do. I have not investigated that, but these Clisterium difficile infections arise a lot of times when people are getting antibiotics that they're taking prior to surgeries. I'm sure now that I think about it, people do get C-difficolitis from IV antibiotics. That's a good point. So, what do we know about the ideal response to helping your gut out when you take antibiotics? This past year, I had to do two courses of augmentin, which is a pretty powerful antibiotic. Kicking and screaming, I did it, but I had a pharyngeal abscess, and that's a no messing around type of infection. And so I'm on steroids and augmenting,

trying to avoid surgery. Luckily, I did. But truthfully, I did nothing after the fact. And I seem to feel okay. Am I just lucky? Or should I have done something? We know that you have an ecosystem in your microbiome. You take an antibiotic that pretty much kills everybody.

And then over some amount of time, you get a new microbiome from the food, the treating environment you're in. And for a lot of people, that new microbiome is the same as your old microbiome. But for a lot of people, it's not. And one of the really interesting things are studies that have been done in kids

where they show that kids who are systematically in antibiotics later on in life are more prone to things like obesity, type 2 diabetes, even things like ADHD, celiac disease. And so there's something I think in those developing years where post antibiotic, you've got a different microbiome than pre-antibiotic. So that's just something to consider on the other side of an antibiotic. If ever you are going to clean up your diet, that's a good time to do it because you're starting with a new blank slate

of your microbiome. And I think what happens for a lot of people is post antibiotic. They're actually starting to feel better and feel well, and then they're craving foods that maybe they haven't been able to eat, and so you maybe aren't giving yourself the best shot at a good microbiome on the other side. I used to believe that taking a probiotic during an antibiotic

course seemed illogical because the antibiotics just going to kill that probiotic. But there have been studies coming out showing that, I think, in particular, this study that's really compelling, showed that if you did a fecal microbiome transplant after taking the antibiotics or, you know, at the tail end of that, or if you were taking probiotics during the time of taking antibiotics, that somehow on the other side of that, you had a, we'll call a healthy gut microbiome, so you're able to have some of these functions that we've talked about, the development of short-chain fatty acids,

and you've got acrimansia in there. And so the idea here is that even though that antibiotic is killing these strains, you might be doing some kind of a seeding at maybe very low undetectable levels that's allowing you to have a healthier microbiome on the other side of the antibiotics. So I guess if I were going to go on antibiotics, I would probably double down on the probiotics that I'm taking. But again, I think on the other side of that really coming in hard and strong with a high fiber diet would be your best chance of reconstituting with really good microbes.

Maybe a naive question, which shows how little I understand microbiology. Antibiotics seem very specific. When you take Augmentin, you're really targeting a certain set of antibiotics. Oh, this really targets this gram-negative strain, and this really targets these gram-positive bacteria. And that's why there are so many different types of antibiotics, right? I mean, it's not just the class of drugs like sephalus borin, you have the first gen, the second gen, the third gen.

And at one point, actually remembered what they all did, which I don't anymore. But the point is there was remarkable specificity. So why is it that if you just take one antibiotic for your skin infection, which presumably would be like a first generation sephalus born? It obliterates your gut microbiome, which by the way, has nothing in common with the bacteria on your skin. But we just described that these are all highly anaerobic bacteria versus these guys that are totally aerobic.

Why does an antibiotic even mess with the gut biome? Yeah, I think antibiotics are a lot more broad spectrum than maybe people or the physicians that are recommending them know. I mean, yes, there's the gram negative and gram positive, but really the name of the game for people who are manufacturing and producing antibiotics is it's actually more useful to create a broad spectrum antibiotic because you want your antibiotic to be the one the go-to of choice from a doctor.

And if it is one that can tackle most different kinds of infection, you're going to become the more popular antibiotics. So, and this is actually one of the arguments in the antibiotic world, which is that antibiotics over time have become more and more broad spectrum. They kill more and more different things, whereas you really want to go in this opposite direction, which is you have a lot more precision around them. There's all these phage therapies and things people are trying to get really specific, but that's new.

Most of the antibiotics out there, they decimate your microbiome. They don't kill everything, which is why you can have these things like they see different infections. Two facents have just quantitatively. You took a gram of stool from me now and looked at how many bacteria were in there, and then you took a gram of stool from me after I was on an antibiotic for 10 days like Augment and very broad, powerful antibiotic. What's the log-fold reduction in viable bacteria? I actually don't know the answer to that. That's probably published in lots of places.

Yeah, interesting. Okay. I actually don't know the answer to that. That's probably published in lots of places. Interesting. Okay. So, what I'm hearing you say is the best thing you can do if you're on an antibiotic is, quote unquote, take advantage of the fact that you're starting with a new team and go out of your way to eat the best possible diet. And we're not really talking much about specifics outside of fiber, are we? What I would do now knowing this, what you've said is I already love eating salad and vegetables

and fruits, so I'd probably go on the all-salad, all-fruit diet for, like, I don't know, a few days. I mean, that might sound dumb, but, like, I would go overboard on those things. Are there other things you would be mindful of either eating or not eating during that recolonization week? I think it's primarily, as you said, these high fiber foods adding the fruits in is important too because those are a source of fiber, but also polyphenols. So we know that polyphenols are beneficial prebiotics for the microbiome, these certain strains as well. For example, we know that polyphenols, consumption, results

in higher levels of acromansia. I mean, I guess just avoid the high fat, high sugar foods. Let's be specific about each of those. What is it about dietary fat or sucrose, fructose and glucose that would be problematic? It's not about the detrimental effects of them. It's actually about what they don't contain. I see. So don't eat McDonald's because it's low in fiber, not because it's high in sugar and fat. Yeah. I guess you're right. The high sugar high fat foods don't tend to be high in fiber, so I was making an assumption

there. But it's really about trying to optimize for high fiber, high polyphenol foods. And so anything that doesn't have that is not feeding. Okay. So you don't have to avoid eating your meat, even though it's not a source of fiber, you tend to make sure you better be eating a lot of fiber. Yeah. And polyphenol. Okay, so you don't have to avoid eating your meat, even though it's not a source of fiber, you tend to make sure you better be eating a lot of fiber. Yeah. And probably, okay, let's talk about artificial sweeteners for a second.

Highly contentious topic. Yes. Recently, along with a couple of my colleagues, we put out a very lengthy piece of content to our premium audience to the newsletter. It's a 15,000-word treatise on all things related to non-nutritive sweeteners and non-sugar sweeteners that are themselves nutritive, so it's a pretty broad piece. Without recasting the entire thing, it's really clear that there's something going on with these non-nutritive sweeteners beyond their caloric or non-caloric impact. In other words, we all understand that sucrose has its four

kilocalories per gram. It's broken down into one part glucose, one part fructose. We understand the metabolism of those things and we understand that if consumed in excess, you have probably some harm beyond just the caloric side, vis-a-vis the fructose molecule and not the glucose molecule. But it's now also clear that under isocloric conditions, high quantities of non-nutritive sweeteners are not entirely benign. And so I guess I'd start with the question of what do we know about how the gut addresses these? If for no other reason, because of the fact that these are very foreign molecules in the concentrations, we consume them. I mean, we consume glucose and fructose for our entire existence. We're just seeing it in a higher concentration now, but probably not

as much of a multiple in concentration as we see aspartame or sucralose. First of all, I'm going to give all the caveats that you are clearly a far deeper expert in all of this and I am. I'm spending hardly any time thinking about it. I know nothing about the impact on the gut, really. Just the observations clinically about what we see in terms of sugar cravings and other repetitive behaviors and metabolic symptoms, which I'm asking, do you think part of that is manifested through the gut?

And there was that very famous paper in nature a few years ago, which suggested that in mice. Yeah, well, first of all, there have been a lot of studies done in mice, and we've already talked about the advantages and disadvantages to getting too fired up about mouse studies, especially in the microbiome. But I think that the data that's out there is conflicting around the impact of these non-nuscious

sweeteners on the microbiome. And maybe it's because of what you just pointed out, which is these things are not all created equal and so by lumping them together and doing these studies that might be causing some of this conflict. I think it's relatively early stage. A lot of studies in the data animals, there are definitely studies which show that they can have a detrimental effect to some of these beneficial microbes. My mind, the jury is still up because I think we don't understand the complexity of these different sweeteners. We also don't understand obviously the complexity of

a microbiome and the adaptation of the microbiome to consumption of these sweeteners. And so, yeah, evolutionarily, maybe these things have been around for very long, but again, because how fast- Because of the rapid evolution. You replicate your- I mean, there's bacteria that can metabolize small molecule drugs. They've definitely never seen before. They've never seen.

So I think that is going to be the name of the game is to understand how does your microbiome evolve to these and how does it help or hurt you and how is that linked to metabolic pathway? My takeaway is all of that and then layered on to that something you said earlier, which is you could take five people who have the same weight and give them the same dose of de-jocsin, and they're going to have five different PKs for those listening who don't understand the term PK. It refers to like the concentration of the drug within their body. In other words, a product of their metabolism.

I would say the same is probably true for aspartame, sucralo, saccharin, all of the above, which is I have seen so many cases of people are trying to lose weight, trying to improve their metabolic health, drinking six diet coax a day, they're saying, look, I'm getting zero calories in here and nothing will budge. Do me a favor, substitute soda water for the Diet Coke for a month.

Let's see if it makes a difference. And the world changes. I don't know what to make of that because it's anecdotal and I don't have perfect control over the situation. So it's certainly possible that when they started drinking all the topo-chico and started ditching the Diet Coke that they were also doing 10 other things that changed. So I really don't know, but I've seen it enough in both directions where it works and where it doesn't work.

I do wonder if there are individual factors where in that individual for whom it becomes a productive change, there's a lack of symbiosis between the evolution of their bacteria in the high-aspirate environment versus the adaptation of another person in the context, my wife drinks Diet Coke. I say that like it's somehow better, but she's freaking loves Diet Coke. She probably has like one every other day. I don't know that that's a higher low dose, but it is. She's in great shape. Yeah.

She's as healthy as a horse. If I drink, I can't stand the taste of it truthfully, but sometimes I'm like so parched and thirsty, I'll drink one. It doesn't seem to cause me any ills, but again, I've seen people, and there's clearly an association between its use, and otherwise. That would be a super interesting study, I think, to do where you basically take a bunch of individuals, it's some baseline data around their microbiome, and then you would put them, you would start them either on a bunch of diet co Cokes or you would start them on a bunch of just soda water and then measure their microbiome over time

And then I do a little washout period or maybe just switch over do a crossover and see what because this is kind of my frustration a lot of the microbiome studies that are done is they treat it the way that we do drugstays Which is you just have cohorts and you're just comparing them to each other. Really, in the microbiome, the person matters. And so, crossover designs are going to give you way more information about what's doing one in that person. And then you can start to draw themes about pathways. But that would be an interesting design to do, because it could be that at the onset, if there was something about these people who could live off of diet coke, and let's assume people prefer diet coke over unflavored soda water, if there was something about their starting microbiomes that enabled them to

lose weight in that way or be healthy in that way, that could be a solution. So like I'll tell you, it was a very interesting study that was done where they took all these people, they put them all in the same diet and everybody has experienced this, you know, to be a physician, you go on a diet, somebody else goes on a diet, one person loses put them all in the same diet. And everybody has experienced this, you don't have to be a physician, you go on a diet, somebody else goes on a diet, one person loses way more weight than the other person. So they were trying to understand is there's something about the starting microbiome that changes the way people respond to,

they just did a regular high fiber diet. And they found that if you start out with higher levels of acrimansia, not to keep going back to this strain, but it really is this keystone strain for a reason. This study, what they showed was that if you had higher starting levels of acrimansia, that was associated with all the metrics

of responding better to the diet in terms of BMI, hip to waist ratio, A1C, weight, all of these things, those people did better. And it's correlative. If they had higher starting amounts, they responded better to the diets. So I do think there's something about the microbiome that can help you or hurt you as you go through these nutrition changes.

To me, a coral area of what you just said is, if your microbiome is suboptimal, which we can define in a number of ways, but let's use one very specific example. If you are completely deficient in or woefully deficient in acrimansia, it is harder for you to have a favorable response to a healthy intervention, and or you may be more susceptible to the downside of less healthy intervention. So on the one hand, you may be more impacted negatively by something like non-nutritive sweeteners, and you may be less responsive to dietary corrections.

Would you agree with that statement? That's what that data points to. Yeah, that kind of a hypothesis. Let's talk about acrimansia then. Going back to it, is the species? There are several strains of it, or when you refer to it, are you always talking about the same strain? Most of the work out there has been done on a particular strain called acrimansia

amucinophila. There's other acrimansias out there, but when people talk and say the word acrimansia, they're really referring to that strain. Okay. How is it grown? You've already alluded to the fact that it's a hardcore anaerobic, and you made some insane statement earlier, which is if even one molecule of oxygen is brought in the presence of its culture, it's dead. Yeah. First of all, can you explain to me, I don't remember enough of my microbiology to know why that's the case.

I understand why an obligate anaerobic survives without oxygen. I understand that biochemistry. I don't remember the biochemistry of why oxygen is toxic. It must be a free radical thing or something. Yeah. They're insanely sensitive to it. Yeah. Amazing. Yeah. Which is crazy because there's someone's oxygen in the air around us, but this is really this compartmentalized body part that we have. But there are these strict anaerobes. You'll see this, especially

we talked a little bit about these microbiome therapeutic companies that are developing drugs. Almost every single one of those companies has had to develop their own manufacturing plant because all these next generation strains that we're talking about that are really in the gut microbiome in the distal colon, they have this same feature, the same issue, which is that you have to create this end-to-end closed system. And so all of us who are developing these sort of next-generation strains and trying to do studies with them, we all ended up having to build our own manufacturing plants in order to solve for this precise problem of the oxygen.

Okay, so were there people out there trying to grow acrimansia without super-superstrick controls? Yeah, for sure. There's a company that is touting pasteurized Acromansia, dead Acromansia as the product that you really want. It's hard to not only grow the strain, but also to maintain its viability on the other side of that as you try to create a product that people can take. There are lots of products out there that supposedly include Acromansia, right? No, we have a whole game of whack-a-mole happening at our company for what we're calling fake armancias.

So these are people who will throw a supplement on Amazon and say, this is Acrimansia. And you can test all these things. Most of them just have lactobacillus or bifidobacterium in them. So they're not even trying to fake it. False advertising. Just false advertising.

Yeah. So walk through the process. So to grow Acrimansia, you have the same vat, but now what do you have to pump into it? Nitrogen? How do you get rid of oxygen? How do you create a no-oxygen environment? You get rid of oxygen by pumping in a competitor, nitrogen. At some point we should have you come visit the manufacturing site. We'll get you all gowned up and take you on a tour. Essentially, it starts with a freezer master stock of the strain. We always go back to the freezer stock because of this genetic component. We have a master stock of the strain, which has been very heavily characterized.

We don't want to just keep growing this thing and then go off of that growth and go off of that growth. I can understand why you would want to start with a master stock because of the profound genetic drift. What do you now do? You grow this in a pure nitrogen environment? You're actually pumping in multiple other gases. Nitrogen is one of the primary ones.

And actually, the mix of gases matter. So that's actually part of the secret sauce of growing the strain. The mix of gases matter. And everything is anaerobic. So in the beginning, when you just have the small stock and you're doing the smaller cultures, you're literally operating in an anaerobic chamber. So you've got the gloves going in this chamber, and you're doing the smaller cultures, you're literally operating in an anaerobic chamber. So you've got the gloves going in this chamber

and you're working in that fashion. And then there is a physical tube that goes from that chamber into an entirely closed then bag, transports out of this bag filled with media and then you're growing it at these larger scales. Everything is an entirely closed lock system and you're pumping in these other gases in order to continue to keep oxygen out.

Because really no matter how close the system is what we found is that oxygen makes its way in. So one of the most expensive parts of this process is the pumping in all these other gases. And if you come visit our site, you'll see we just have walls and walls lined with these tanks of these other gases that are getting pumped in. And then after that, again, everything through liophilization has to remain anaerobic. And then once it's freeze-dried, now it's in a stable state. So I could leave agramancia powder out on this table for weeks and it would be fine. So it's really just leading up to that freeze-dried state. Wow.

So freeze-dried agramancia, I guess that kind of makes sense because a capsule presumably still allows for diffusion of oxygen. Absolutely, and the freeze-dried state, you basically put them into a dormant state, so they're not metabolically active at that moment. And it's so interesting, so you don't have to keep them in a freezer at home. A fridge is good enough, even though they had to be cooled to a much lower temperature, presumably to be freeze-dried. They can be warmed too as much as a refrigerator, which is probably what 38 degrees or something.

Yeah. Well, actually, acrimansia, the single strain, is room temperature stable, so you can actually have those pills at room temperature. And that still doesn't permit enough heating. Yeah. The body temperature is what's necessary to heat it. Even then, when you consume acrimansia, it's the hydration that's actually your big problem once you're in a freeze-dried state.

You're basically trying to keep that guy dormant. And so, once it's freeze-dried, and when you buy our bottle, you'll see we have desiccant packets in there. We have desiccant line bottles, and we do our clinical trials. It's all about keeping moisture out. Now once they're in a freeze-dried state, now once water enters the game, that's what brings them back to life, and then they die, because now there's oxygen.

So, once they're in their capsule form, some people tell me, like, oh, I threw out that little packet. I'm like, don't throw that out, because that's what's going to help you keep these guys stable. Wow. So, what's that process take in terms of time? The time varies for each of the different strains, and obviously, also the concentration and the number of bottles that you're trying to make. So, varies for each of the different strains, and obviously also the concentration and the number of bottles that you're trying to make. So the people who do this, obviously,

this is a high-touch industry. They were doing what before this industry. This is something that's done where? You mean before do manufacturing probiotics? Yeah. Probatic manufacturing has been happening around the world for a very long time. But using this high-intense anaerobic process.

Actually, we built this thing from scratch. We had PhD scientists and microbiologists who were taking these small-scale methodologies and then trying to figure out how do you grow them at larger scale. In fact, our manufacturing plan is in San Francisco because that's where all of our PhD microbiologists were living, you know. And so we actually have a pretty young team of people that are coming, almost some of them straight out of their postdocs.

How are you doing the flow cytometry on the anaerob without introducing oxygen? The flow cytometer is in an anaerobic chamber. I mean, I used to do flow cytometry every day. They're huge. Yeah. We have custom built anaerobic chamber. I mean, I used to do flow cytometry every day. They're huge. Yeah, we have custom built anaerobic chambers for all this stuff. Well, how does the person go in? Are they just putting their arms in?

Yeah, usually these gloves, you stick your arms in. But to prepare for the facts, you're mixing the antibodies, your putsin' around, you're rinsing, you do all that through these little glove chambers. All of it is done through these gloves. That's hell on earth. And then there's a little box on the side that you open it up, you put rinsing, you do all that through these little glove chambers. All of it is done through these gloves. That's hell on earth. And then there's a little box on the side that you open it up, you put your thing in there,

you close it, you have to de-oxygenate it. It's like a submarine. It's like a submarine, the double chamber. It is. I think people get pretty adept at using these gloves. I'm terrible at it, but they can do really fine work with them. Wow. Okay.

Aside from the fact that Acromanceia from an epidemiologic standpoint always shows up. How long has your company been selling in Acromansia probiotic? We've only been selling it for a couple of years. I mean, we really spent about a decade doing dev work and R&D work and preclinical and clinical work. I wanted to make sure that thing did what we thought it was going to do. Are you comfortable saying how much money you spent on that R&D? A lot.

Our company has raised $150 million and we've only had products in market for about two to three years. So, it was a heavy lift to build a manufacturing plant, do all the R&D work, funding pre-clinical and clinical work, we fund other PIs to do work with these strains. I mean, I think if you want to build a product that has real efficacy behind it, there's a lot of blueprint from pharmaceutical drug industry that we've adopted because the incredible rigor that they have around developing products of efficacy. Yeah, and your industry is not one that's known for rigor.

We'd be putting it mildly. Yeah, and this is a really pivotal moment for our company when we really decided to sell to consumers. So we'd always had this idea that the microbiome is really interesting because it's mutable and you could develop things that could help people. There is a lot of evidence suggesting through these ethical microbiome transplants

that you can actually help people improve health through a microbiome intervention, and that if you followed a drug development rigorous pathway, you could actually identify the subset of that fecal microbiome transplant that could help people. And if you could very hard to manufacture them, you could really have a product with efficacy here. Now, how do you bring that to market and how do you choose to commercialize that? We always believe that because these are grass, these are naturally occurring strains,

that you had an opportunity to bring this directly to the public that would allow you to really democratize the availability of it. So if we launched Pendulum Glucose Control as a drug, we would only be able to sell it to doctors who would prescribe it to people who had type 2 diabetes. Anybody who's aging should be thinking about how to help their body metabolize glucose better.

And so we really felt like there was this natural product, things that could deliver efficacy that was measurable that would be important, and then enabling these products to be available to everybody as opposed to only through a prescription that meant that the consumer market was really the way to go. Also, a lot of people learn information about health on their own. I mean, Dr. Google is the most famous doctor on earth. And so we

really wanted to bring it directly to consumers. The downside to that, and we've talked about this too, is it's really hard in this particular space to elevate it. I mean, everybody is a marketing genius that's selling a probiotic. There are people who make you feel like what they're selling is super innovative and you've never seen anything like this before, but when you really look at the ingredients, it's literally the same thing that everybody else has been selling.

And because a consumer is not gonna go reclinical trials or necessarily compare your ingredients to ingredients and all the other labels or do all that leg work, it becomes really different game to play where you're trying to deliver something that provides meaningful health solutions for people, but you're also having to play the marketing game of how do I convince people or use proxies to help them understand this works because you're not selling to a bunch of doctors, you're selling to regular people.

Well, even if you are selling to doctors, I mean, quite frankly, it's complicated. I mean, it's not like doctors would necessarily understand this. I struggle to understand this. Let's explain what acrimansia does to control glucose. You've alluded to this a couple of times now. You've alluded to a product called glucose control. Let's kind of go back to the science of this. There's a clear and obvious correlation as to why acrimansia is beneficial,

but that doesn't explain mechanistically what it's doing. Do we have an insight into why acrimansia is beneficial, but that doesn't explain mechanistically what it's doing. Do we have an insight into why acrimansia is something that lowers an individual's blood glucose or response to a glucose load? Yeah, so we'll do a deep dive into acrimansia and what we know today, knowing that we're going to learn all kinds of other stuff, but at a very fundamental level, this is primarily through the GLP1 pathway. And just to get people excited about that, if you're hearing this and you've heard of OZMPIC, you've heard of GLP1. OZMPIC is a memetic of GLP1.

Yes. What we know about acrimansia, there's three key things that we know that it does that kind of result in this. So the first thing is that it has a surface protein called amyook 1100 that appears to be able to bind to TLR2 receptors that helps stimulate these L cells. So maybe we'll take one step back, which is that you have these L cells in your gut microbiome, at the lining of your gut microbiome, and L cells are the cells that secrete GLP

1. So, a lot of people don't know this. Your microbiome is really the guy secrete GLP 1. So, you have this beautiful system where you eat food, your microbiome metabolizes it, and right there are these cells that respond to that food by secrete GLP 1. So, acrimansia has this surface protein called amyook-1100. It also secretes a protein called P9, and P9 binds to these ICAM-2 receptors in the L cells, which are also known to stimulate them to produce GLP1. And then the third thing that acrimansia does is it's able to produce a short-chain fatty

acid called propionate, and propionate is upstream of butyrate. And so there are a bunch of strains which can take propionate, converted into butyrate. Butyrate binds to G-protein coupled receptors 42 and 44, also in these L cells, and stimulates GLP1 secretion. And so these are all ways in which acrimansia can stimulate GLP1 secretion. And by the way, there are only two strains shown to be able to directly stimulate GLP1.

One is acrimansia musinophila. the other is clostridium butyricum, and they actually act together because clostridium butyricum, as its name indicates, is a butyrate producer. And then GLP1 has multiple benefits, one of which is that you know all this stuff way deeper than I do, but helps your body know that to secrete insulin, to help you metabolize the sugars that are in your blood after eating a meal, but it also has this other benefit, which is what is becoming really popular now, which is that induces the tired. It makes you feel full. And it does that in two ways that are not totally understood, but one is that it slows

your GI transit, which gives you a feeling of fullness, but the other is that it has some kind of a neurotransmitter mechanism that allows you to feel like you don't have cravings. You're full. It makes it a very powerful small molecule, and so what we're doing that's pretty distinct from what these small molecule drugs is doing is the natural way we're giving you the upstream bacteria that's enabling your body to produce GLP1. The result of that is that your body will produce GLP1. If you eat in is that your body will produce GLP1. If you eat in food, it'll go back down. It'll produce it again after you eat in food,

but it will raise your levels of GLP1. So you get these benefits of lowered blood glucose, reduced food cravings, lowered A1C, and the body weight impact. Okay, a lot of want to unpack there, starting with just a quick summary. So you eat food, provided you have a lot of acrimansia and you're feeding it the right food, a couple of things happen. It sounds

like one of them is not at all related to a production or a secretory product, but rather just this surface receptor. I think you said it was the AMIC 1100 or something like that. Yes, it has a very fancy name. Yeah. And that surface protein by itself just stimulates L cells in the enterocyte. We know that that makes GLP one. Yeah. And I think that pathway is probably the least well understood. Shortching fatty acid in the P9 protein have had, I think, a lot more work done on that.

So it's the Crete's P9, it's the Crete's propionate. Propionate gets converted to butorrate by another bacteria, which ones? There is a class of clistridial strains that will do that secondary conversion. And clistridium butyricum is sort of the most well-studied of those. Got it. What about difficile?

Is he doing anything good for us there? Actually, don't know. We haven't studied whether, and maybe people are afraid to bring that into the lab. Yeah, yeah. And what does P9 do directly stimulates L cells? Yes, it binds to this ICAM2 receptor on the L cells and stimulates them. ICAM receptors are receptors that are usually involved in, God, if my memory serves me correctly,

like kind of an immune response, right? Like an inflammatory response? We have not done studies around that, but for sure, that is a area of heavy interest. Yeah, by vague recollection, I mean, we're literally going back 25 years is that when a person is, for example, septic, you have all of these ICAM modules that lead to vasodilation, leaky capillaries,

secretion of monocytes out into the peripheral tissue as macrophages, but it sounds like this is a distinct process here. And maybe I should say to, like, I don't know how much of this is tied to that process, but not only does acrimensis stimulate these L cells produce GLP1, but acrimensis has another role, which maybe is very much tied to this, which is that it helps regulate the mucin layer. And so that mucin layer turns out to be super important for, if it gets too thin,

or if it's too thick, it's a real part of the so-called leaky gut, or GI issues that become a result of not having enough acromansia, is really that you have too thin of a mucin layer. And so you start to get the ability for pathogens to infiltrate, and then also obviously for these molecules to citrate out. And then tell me what's the fate of the butyrate made of the propionate? It binds to these G-protein coupled receptors, and that's also what stimulates the L cells to then go release GLP1. Got it. So there are multiple these small molecules that are the signaling molecules

for these receptors on the L cells. So all roads point to more acrimansia, more GLP one, what differs from someone taking ozemic is they just have a mega high dose of GLP one all the time. And what you're talking about here is, as you said, kind of a waxing and waning dose of GLP one that is more physiologic because it comes with your meal. That's right. So one would not expect this type of intervention to produce the amount of weight loss you would see with carpet bombing somebody with GLP1. Absolutely not.

And one of the other big differences is that the GLP1s are injected. So it's going right into the bloodstream where it's a microbiome effect. And to your point, you get the waxing and waning, more physiologically relevant, but it's not going to be the same as just hammering a bunch of GOP1 strain to the bloodstream all the time, all day, all night. So now let's talk about the data. So what was the first study that demonstrated that acrimansia could play a role from an intervention perspective from impacting metabolism vis-a-vis blood sugar?

Yeah, some of the earliest studies are really done. I mean, so for example, I was discovered by Dr. Lee Kaplan over at MGH, and he's a very ashiic surgeon, and so his initial interest was what's happening to the microbiome we do this very ashiic surgery and came to really be one of the first people to really look at these microbes and discovered that acrimansia appeared to be associated with or inversely associated with obesity and then started doing these in vitro studies to figure out like what is it doing. The work really has to go credited back to him in the early 2000s. I want to point out one of the observations that came from his work which is a really remarkable observation which is at least as far as the Ruanwai gastric bypass which is a really remarkable observation, which is at least as far as the

Ruinwai gastric bypass, which is a real organization, a reorganization of the plumbing of a person's gut, that you took a person who was obese with type 2 diabetes, and you do a Ruinwai gastric bypass on them. And within days of surgery, their glycemic control improves even before they've lost weight. This is a very important point because nobody would find it that surprising if a person after a gastric bypass loses 100 pounds and their diabetes goes away. That would be a O-shocks, of course, moment. It's why does their diabetes resolve in the days following surgery when the real weight loss hasn't started? And there have been lots of suggestions to this.

One could be the fasting. You're not eating anything a day before surgery or a day or two after surgery. That's a pretty significant fast. That depletes all of the glycogen in the muscles and much of the liver glycogen is that enough to kickstart them. But it's hard to ignore that you're also completely changing their gut biome by re-plumbing their GI system. So can you say more, because I'm not familiar with those data in terms of what they saw as far as pre and post surgical gut biome? I'm also not an expert in these surgeries.

They have been a source. A lot of publications have sort of used them to try to understand what are the key gut microbes out there. And I don't think it answers this question of why do you see such an immediate response sometimes hours after the surgery, just thinking about the time length of these different pathways to have effect. It's most likely something hormonal, but I don't think we have the answer to that. It's hard to imagine it's not something related to GLP1. Yeah, so I don't think we have the answer to that specific question, but definitely that was where

acriments who was born out of. They do these gastric bypass surgeries in, there've been a bunch of animal models. This was some of the very early work really just trying to lay the foundation for the fact that your microbiome could play a role in weight loss or weight gain. And so they would take these microbiomes before and after the surgeries. And then they would put them into these mouse models. And they would show that you could change the metabolism of the mouse. It's been really hard in the field to take that data, that fecal microbiome data, where you feel like, oh, there's a glimmer of hope here, and to distill it down to, what are the things that are actually doing that?

That's the big challenge of the field. We're not there. So when you say, like, oh, what did they find? It was, well, they found that the whole Gammish could change things, but we don't really know what in there is doing that. Yeah. One hypothesis is that acrimansia is doing part of that at least. Yes.

And so the easiest way to test that would be to give people acrimansia. Okay, so you guys did an experiment like that. Who did you do that in collaboration with? I'll point to the clinical trial that's published in BMJ because I think that was probably the most rigorous trial that we've done. And then I'll talk about the downside of running too many trials in things that involve humans. But in that trial, we didn't just have acrimancy.

We actually had it in combination with four other strains. The idea here is, as I said, acrimancy can produce propionate, but without the help of another strain, it can't produce buterate. And so we had this idea of, let's include primary and secondary fermenters. Let's not make acrim say the only primary fermenter. Let's add some more in there. And really thinking about what is the team members that you'd want in here that can metabolize fiber and debuterate, trying to optimize or buterate production. At the time that we did this study, nobody even knew what P9 was or AMUK 1100. We just knew that we

thought it was only playing a role in the mucin regulation. We didn't even really know that it was able to stimulate GLP1 even at that time. So we really thought it was these butyrate producers that are doing that. Which are the clostridial strains? Yeah, which are these clostridial strains? And then also this bifidobacterium and phantastrain,

which is also a primary fermenter. So we basically did a placebo-controlled double-blind and randomized trial where we had three arms. One was placebo. One was this full formulation of all five team members, and then the third one was a subset that did not include acrimansia. And these were in people with type 2 diabetes, initially we wanted it to be people who just had type 2 diabetes but weren't on any medication.

It turns out that's like almost impossible to find. Yeah, it's hard study to find. Yes, so then we said, okay, we'll have it with people on Metformin. And anyway, a lot of people are on Metformin, so if your thing is gonna benefit people, it ought to work on top of Metformin. Recruiting in the clinical trials is hard. It takes a long time.

We were start upcoming, you were like, we're gonna run out of money, if we don't expand this out. So then we started including people on sulfonuria. So finally, we get all the people in trials, still a pilot trial, 76 people, cross these three different arms. And what we found is the people in the trial, still a pilot trial, 76 people, you know, cross these three different arms. And what we found is that compared to the placebo group, the people who were on the full

five-strain formulation over a 90-day period saw their A1c go down by 0.6 and their blood glucose spikes go down by 34%. Measured how? That was done using a pretty old school oral glucose tolerance test. They literally came into the clinic, they got their sugar, and then they just had blood drawn every 15 minutes for two hours. And the reason we did that rather than a CGM was because

we wanted to use all the gold standard traditional methodologies. We don't want anybody to think that the data was weird. So hemoglobin A1C came down by 0.3%. There's a pretty big drop. Six. 0.6%. Compare a pretty big drop. 6.6%. Compare to placebo, yep. Compared to placebo in 90 days.

And peak glucose level fell by about a third, you said. Area under the curve. So the entirety of the area under the curve, yeah. Okay, fell by about a third. Yeah. Got it. And that was placebo to five strains. Accrimansia plus four strains to do the conversion of propionate into butorate.

We also have strains that do the redundant function of acrimancia, which is they'll do that primary fiber into the propionator acetate. I see. Okay. And then how did the group absent acrimancia do? They had some efficacy, but it wasn't like the five-strain formulation. So my co-founder and biostatistician would say that was not statistically significant, and the data is published, so anybody can go look at it. I would say it looked like it was in between the placebo

and the five-strain formulation. But it could have been very underpowered, and that's why you didn't see a difference. It also could have been very underpowered yet. So this was thinking about this now, not as a scientist, but as an entrepreneur. This was a do-or-die moment. There's going to be a binary outcome here. Either we were going to be a company on the other side of this or we were probably going to close up shop because it was all in on this trial.

What year was that experiment done? So this was maybe 2018. We founded in 2012. All the stuff leading up to this was we 2018. We founded in 2012. All the stuff leading up to this was we had identified the strains. We had to figure out how to manufacture them. So six years of basic science. Six years of basic science leading up to this trial. And I was already starting to think about, it's very rare that your first clinical

trial out of the Gates works. This is why we did the three strain and the five strain. Yeah, why didn't you just do the placebo versus the five strain? Because acrimansi was so hard to grow. Oh, so you were hoping to see that the four strain would be great, and you wouldn't have to grow acrimansia. Yeah, we were hoping we didn't need the diva in there, it turns out we did. So what's interesting scientifically, but would have added too much overhead to the study,

and maybe it's irrelevant if the other four are not that expensive to grow, is would acrimansia alone? How would that have performed relative to the five-strain pot? Yeah, of course, if we had been able to do that subsequently, but I will say that lots of people have been doing work with acrimansia solo, and I put Vegas odds down that by itself, it's probably not as impactful as a formulation. Because you don't have the assistance in converting propionate to buterate. Exactly.

And then sorry, dumb question. I think you already addressed this and I've forgotten already buterate is stable. It's a short-change fatty acid. Why aren't we just all mainlining buterate? There have been a ton of preclinical and, of course, in vitro works showing the massive benefits of butyrate across a bunch of different states, including metabolism. And yet none of them is translated into humans.

Even when we try to do these animals, these clinical trials, they've never looked the way they did in the animal study. The animal models are like, blow your mind, blow you out of the water, and the human trials really don't show that. And I think for this particular pathway that we're talking about here, it's a localization problem. So earlier I mentioned that butorate is source of energy for all these calonic cells.

The issue is when you're delivering butorate all along that track as this butorate is basically being absorbed by all these different cells. And where you need it to get to for this pathway is to the gut lining where the G-protein coupled receptor is sitting, bind to that G-protein coupled receptor. So it's not that acrimansia just makes buterate. It's that it delivers it to the place

where it needs to be. Exactly. Literally, the physical proximity is what's enabling this to work. Wow. Let's go back to actually something you said a moment ago, which is this trial was done in people with type 2 diabetes.

Average hemoglobin A1C coming in was what? Roughly. These are kind of early diabetics. They're in the sevens. Yeah, they were in the sevens between 7.5 and 8.2. I think was the range for all the groups. I have to go back and double check that. Is there any sense that I know you didn't do this in the study because you said it was

only 90 days, but is there any plan to redo this to find out what happens if they had stayed on for a year? In other words, could you take someone with type 2 diabetes and actually knock 1.2% off their hemoglobin A1c, which for many people would now take them straight out of the diabetic range? Yeah, we have multiple ongoing studies. Maybe this will get back to kind of the personal story of the entrepreneurship range. Yeah. We have multiple ongoing studies, and maybe this will get back to kind of the personal story of the entrepreneurship, which is that on the heels of this study, we are like,

holy shit. We made something that works. Now I got to go figure out how to commercialize it, but before we do that, we should probably replicate the study, or maybe we should do them, you know, in parallel. By the way, did you get any pushback from your investors on that? Doing another study? Yeah, because like, if you're wearing your scientific hat, that's the obvious thing to do, because scientists always want to replicate things

and make sure and add another question. If you're an investor, you might be like, are you crazy? We just got the answer. It's the best possible answer. Don't ever ask another question. I mean, there's a reason virtually no supplement companies will ever, ever, ever do a clinical trial.

They don't want to know the answer. It's true. It's true. Well, there's two things. First of all, we have been super fortunate in having investors that believe in the underlying premise of our company. And the underlying premise of the company is that the supplement space is riddled with tons of products that don't really do anything.

And therefore, it's very hard for anybody to own market share. It's actually a highly fragmented space. Sometimes you get a player that comes in, they take market share because they've done something fancy on the marketing side, but then somebody else comes in and takes their market share. You just sort of see this game happening. Our underlying premise was if you could deliver something

that actually helped people in measurable ways, you could take that whole market over, because now you've created something that works. Well, in order to know if it works, yeah, you have to have a study and you have to make a claim. So you can now make a claim with a product, that's a very big deal. Are there other probiotic companies out there

that can make claims? No, this claim around lowering A1C and blood glucose spikes. You own that claim. Yes, we are the only ones that make the illest somebody else could do a study and make that claim to the bad. Yeah, but currently nobody else can do that. No, nobody else does. The other reason I think our investors got behind it is, first of all, they understand

that we're trying to build products that can change the lives. And we have investors that have gotten behind game-changing, category-creating products in the world. So Apple and these sorts of companies that maybe didn't have a predecessor to them. And so that's a very different risk tolerance than biotechnology. Biotech is the graveyard of some great investors. It's true.

I don't know to say they're behind this mission that the thing has to work. And so therefore, you've got to have these studies that work. And I will say though, I have learned by having products in market and enabling people to run their own studies on themselves, especially in the context where nutrition and your ecosystem and your microbiome are still pretty much a black box of the scientific community. We've been able to garner so much data from our customers. We're just sharing this stuff voluntarily with us in order to be able to do better and better product development.

So I am actually now a believer that the clinical trial is important because it gives you a very controlled environment to know whether there's a there there, but unless you can change behavior in the real world and have people actually experience benefit that they can come back and tell you about, you have a made a product that's very interesting at all. And so I think that that part is important. The other part to this story that's important on our strategy is that because we have this clinical data and because a consumer may not be able to go read a paper but a doctor can, that we go through healthcare professionals, we go directly to them, but then we also

go through them to help create credibility or knowledge for consumers. And so if you want to convince a bunch of doctors that the thing you have works, they're going to see more than a pilot. And they're also going to want to see it run by somebody that's not yourself. And so we immediately launched these studies and then COVID hit. And every study one by one got brought to its knees because people with diabetes are more prone to COVID complications that that came out a little bit later on. But people were getting COVID. And then you're like, well, I don't know what this COVID

is doing to the microbiome. And then people were totally not adherent to any protocols. They couldn't come into clinic. So one by one, we lost a lot of money on trials that we were trying to follow-ups on. And then we just decided, my chief medical officer said, all right, we have to wait until everything is cleared out before we even start another clinical trial.

Because we're not going to lose another dollar. And so literally just in this last year, 2023, is when we felt like there was enough of a handle on this that we could actually start doing clinical trials. But we've done it in a different way, which is now we sort of built up some momentum. So it's all third party PIs and academic and clinical institutions that we're just giving free product to to run these trials. So we'll get that data. I do think it's important if you want to get the medical community behind you.

And I think that'll be a differentiator. That means they have to apply for their own grants to fund the study. They're just having a significant part of the cost taken out of it, which is the drug. But that's good. So there's interest. There are PIs out there who are like, yeah, I would happily go and seek the grants to do this study, especially given that I can mitigate the drug cost. Yeah.

And the government is putting out, you know, NIH grants. There's quite a few of them centered around microbiome interventions. So how many such trials are underway right now? Right now we have about a dozen around the world that are happening, not all of them are for type 2 diabetes. We actually have several, were investigators have their own hypothesis about what these trains are doing. So for example, we sort of talked about the gut brain access.

We have an investigator that's been funded by the milk and institute that is looking at the role of acromansia in bipolar disorder. That's kind of amazing if you really think about it. So you've got basically a dozen PIs around the world on the basis of one relatively small pilot study that was very well done, published in a very reputable journal, and that alone has generated that much interest that they're finally willing to study a commercial product. Well, I don't think it was just that study. There's like over 3,000 publications on acrimansia that people around the world have been publishing on, you know, mostly academics.

But those studies weren't intervention studies. No, that a lot of them are correlative studies. Right, but that's my point. What you did is quite unique in that you finally bridged a gap, which was correlate, correlate, correlate, but until you can test an intervention under blinded, randomized conditions, you can't know if there's an associative link, and you've at least suggested that. Absolutely. And I think that you're right.

The fact that we can make it and that it was able to have this efficacy does give people some belief that, okay, well, if I want to go test an intervention, these are the people that have made it. Right. They could never invest in the CapEx to go and make it. No, I mean, our manufacturing plant, it costs us $10 million to make that plant. Yeah. Okay. So how many products do you sell today pendulum cells? The product that was tested in that trial is that the product called glucose control? Yes, so pendulum glucose control is marketed for people with type 2 diabetes Lower say one C lowers blood glucose spikes and it is the exact formulation that was in that clinical trial And do you still use that same quality control metric of actually doing flow cytometry on those products?

Yeah. I take glucose control. If I looked at that bottle, which I can't say I've looked at and remember, it doesn't say anything about CFUs on the back. This is AFU, active fraction unit. And what does it report it in? It's in the number of cells that appeared in that active fraction. So it's still going to look like, I think it's like 10 to the 8 or 10 at night for most

of the strains. Got it. Okay. What other products do you sell? We released pendulum glucose control. And I would say one of the hallmarks of our business that we've been very proud of is the repeat business. So we get a lot of, once people try it, they really stick with the product.

We actually got a lot of feedback from people. We were getting amazing reports of, oh, my A1C is lower, my black glucose pecs have lowered. When we launched our first version of glucose control, we also offer people free A1C testing every 90 days and a nutrition coach, which we had a team of registered dietitians. So we were like, this is gonna be a solution, right?

We're gonna give you your nutrition coaching, we're gonna give you the test that's gonna type. It's working. You how did you offer that for free? You didn't wanna make money, I'm guessing. We don't offer it now. Somebody even said, oh my gosh, everyone should go buy this product right now

because these are non-scalable. So like at some point, they're not gonna offer these things. And I laughed when I saw that, I was like, oh, you don't know, but then eventually I realized, though, that's true. I think that it's still important for us to offer nutrition information. It just doesn't necessarily have to come through a registered dietitian and a personalized 101 coaching. And maybe AI will help us get

there faster than we would otherwise. And what other things did you hear from customers? So did you see in the real world comparable reductions in hemoglobin A1C? It was all higher. The reports we get back were higher A1C drops, higher blood glucose drops. Do you have a sense of how much selection bias was going into that? Were there a good number of people saying what the hell, like it didn't get better? No, actually. Moreover, I would say you might say, well gee, those people just didn't tell you, but you can see it in the numbers.

You can see it in the return purchase rate. And pen just in glucose control is $165 a month. That's a real out of pocket expense. I don't really know the space that well, but how expensive are the top-selling probiotics? Normal probiotic is 20 bucks a month. A premium high-end probiotic is 49 bucks a month. So 165 is way out there.

I think you've explained why it has to be that expensive. I won't ask you what your margin is on it, but I'm guessing it's not that high given your manufacturing process. Well, what's even worse is that you're teasing me about offering this test and this nutrition coaching and how did you do that? We were not only losing because of those, but we were actually losing on every bottle we sold because it was costing us more to make it than the 165 we were selling for. So it was all kind of a hot mess out of the gates, but I think what we believed was that we knew that as we scaled a lot of those costs to go down. So just like everything else, if you're buying 10 bottles

versus 10,000 bottles, the same bottle costs you a lot less. Can you currently make a bottle for less than whatever that cost is? We now do make it for less than what we're pricing at, but we also have to ship it cold. So there's also that transport. Yeah, that's an important point. It arrives, you get three bottles,

which is a three month supply. It arrives in a nice pack, in a big box, just like if you were to repath or something, like a PCSK9 inhibitor, same thing. Shows up cold, you got to put it in the fridge right away, can't travel with it. I mean, there's some logistic challenges associated with these things.

There is definitely complaints. But I think what we wanted to do was to give the product the best chance to success by giving people these tools to be able to help them measure and then also, a lot of people don't even know what a high fiber diet is supposed to look like. And so just giving those tools to people. So we got all this great feedback. Efficacy was higher than what I'd even seen in the trial, but we also got a lot of complaints. Hey, man, how is this $165 a month? Why is it my insurance covering it? Why does it have to be cold? I can't travel with it.

What am I supposed to do in the time that I'm traveling? I don't have type two diabetes. We kind of took all that and said, well, let's just run some marketing tests on a lower dose version. That had huge uptake. And so we launched a product this year called Metabolic Daily. It's literally the same thing as glucose control, but just at a lower dose that allows us

to get down to this price point of $49 a month, which feels a lot more manageable for people when they're thinking about, especially if you don't have type 2 diabetes, but you want to help your body metabolize sugars and carbs better and things like that. That product still has all five strains in it. That product still has all five strains in it. Yeah. So it needs to be refrigerated. All of our products really should be refrigerated. That does maintain the viability for longer.

Those two products, and then we also release the single strains that are components of these products. So we had a lot of people who came back to us and said, hey, I'm buying your pengel and glucose control, but all I really want is acrimansia. I just want that one strain. You're sort of like, who the hell knows what acrimansia is? This is just like some really educated people coming to us and it's not a real market. So we did a market test. We made a

thousand bottles of acrimansia. We literally just called it acrimansia and we put on there like that's for gut health. We made no claims about it. People were coming to us for wide variety of reasons of why they wanted acrimansia from metabolism issues to GI issues to neurological disorders. So we were like, we'll just sell it as just acrimansia. That's the value prop. We made a thousand bottles in less than 10 days every bottle was gone. And that's an expensive product because it's acrimansia. One of the hardest things that is for me. And so that is a product. It's one of our best sellers. It's just pure acrimansia. And actually a lot of practitioners use it because let's say they ran a gut microbiome test

to persons alone at acrimansia. They just want to give them the minimal viable product to boost this strain back up. But we're really on the outskirts of knowledge here. We really don't know what it's doing by itself. Well, we know that it's secreting some of these proteins. But in terms of outcomes, that's more of a stretch, right? At this time. I would say that what we've learned from having it in the market is that people are getting

a variety of different measurable impact from it. And one of the most interesting things that we learned is that we had a bunch of people telling us, actually the reason why I stay on it is because I don't have any more sugar cravings. I went to the Christmas party and didn't eat a single cookie and I'm usually the guy sitting at the table eating all the cookies. We started to see this theme of sugar cravings and as we talked about earlier, one of the hallmarks of GLP1 is improvement in satiety. We just did a little pilot study ourselves. We said go on this product for 90 days and do this diagnostic test on cravings. Was it a blinded study? Was there a placebo arm? No, so this is a pilot. This is a proof of concept. And so what we see is that people have a significant reduction in these food cravings. And so what that's

now being parlayed into is funding a clinical trial to look at that. And so this is kind of where I think the consumer space is a really interesting place to look at that. And so this is kind of where I think the consumer space is a really interesting place to play because rather than going all in with millions of dollars on a clinical trial based on a hypothesis in your head, you now can put products out there to people, hear back what they're experiencing. Yeah, generate hypotheses from customer data.

Get proof, concept data, and then you feel like, okay, a priori, I have some sense of things going to work. And you could even get really sophisticated and say, okay, well, maybe I'd like to know what are people starting microbiomes or what are their diet or what are their lifestyles or the demographics to try to understand who might this thing have the most efficacy for. Now, you also haven't done the study I'm guessing where for those, look at your pilot study or your 76 person study, it would be very interesting if you go back in time if you had the funding to look at what the Constitution

of the Gut Biome was prior to the study after the study and if that predicted response. In other words, is the greater the deficit of acrimansia at the outset of the study, a predictor of a greater response upon normalization? Because even though the average Heminglow A1C came down by 0.6, there must have been people for whom it came down by over a percent, and people for whom it only came down by 0.2. So do you have a sense of what that relationship is?

Yeah, well actually in that trial, we did, we got four stool samples from people during the child. So one, yeah, baseline, I think was like 30 days in, after the 90 day mark, and then we did a wash-up period. So you went for a month without taking anything and then we got a stool sample. And I'll tell you this, we racked our brains about how are we going to get people to provide four stool samples? I mean, this is like a real ask.

We wanted the whole sample. So we literally was like these cool whip. That's what we gave to people. And they had to literally shit in a bucket, put it in their own freezer until they could get to the clinic and everything had to stay frozen. And so we had a hundred percent compliance. We had people who dropped out of the study who were still sending us their shit. I mean, it was amazing how much sample people were willing to share. By the way, I remember my brother got back from like, I don't know, he was somewhere in

Africa or something and he had some awful GI bug and eventually like he had to collect a stool sample. I was like looking at his fridge and there was this bag in there and I was like, what's that? And he's like, I asked you some shit. And I was like, no, seriously, what's that? He goes, ah, it's just some shit. And I was like, dude, if you don't want to tell me that's fine, but you he's like, no,

no, that's what it is. That's literally what it is. Yeah, exactly. People are sending a shit. And the reason is, because as I said, we were in early stage startup, I was like, the chainsaw of this clinical trial isn't gonna work.

I gotta know, we have to have this microbiome information so that in my head, I thought, for sure, if this thing doesn't work. I wanna at least know it didn't work. Exactly. Did it get delivered or not? And so we really were wanting to see the presence and absence of our strains.

But to your point, we also really want to understand, could you do this predictive modeling of who would be better or worse responders? And we couldn't. It turns out that could be that the end is too small. It could be that we don't know enough about these individuals, but the microbiome alone or even the levels of acrimancy alone are not enough of a predictor. They're not enough of a predictor of response. And tell me this, did anybody who took acrimancy have a significant increase from pre-settied to post-studied during the 90 days, and then what was the fall off from 90 days through the washout? Every participant in the study showed an increase in all the strains.

And so that was very rewarding, because we invest a lot in the encapsulation to get the thing delivered. So that was a great proof of concept. We actually managed to deliver something that in theory is undilivered. Exactly. Deliver the goods. And then the wash up period was 30 days. So after the 90 days, there was a 30-day washout period, and most people lost the strains after that period. But there were about 15 to 20% of participants who were able to maintain their acromanceal levels even after not taking the pills anymore. That was exactly about to ask you that question. Is there any chance that those are the people who just consume the best diets and ate the most fiber after?

So, this is another decision that may be in retrospect. We didn't do diet logs. And the reason we didn't do diet logs is because I am told that people are liars. When you do diet logs and clinical trials, what you get back is all the day somebody was good and then big gaps of missing days where they ate something terrible or it is something bad.

I almost think of doing a diet log here where the only thing you're asking them to log is fiber. And you give them a really clear sense of fiber. And it's just all we're doing is counting grams of fiber per day. Take a picture of everything you eat that is one of these things on this laminated card. Every time you eat a carrot or a celery or this or an orange, just take a frickin picture of it and tell me how much of it you ate. And that's it. I don't care how many calories, how many diet coax, I don't care about anything, just eat all the cookies you want if you want. I just want to know this.

Yeah, I wish I'd met you when we were doing the trial design. So we didn't do that. What we did was we asked people, what are you? Are you an omnivore or are you a vegetarian? We have all that. There's a appear to be a correlation with that. And then we asked people, please don't change your diet. The whole point of this study is we don't want people to have to undergo a behavioral change in order to see an improvement. It should be just the microbiome

intervention. But of course, we don't know if people change their diets. I think what we want to tackle now is a much more direct thing, which is to say that you can deliver people meals, and they don't know to eat the meal every day. But basically, if you say part of the trial is, you're going to be on the product, and then we're going to deliver you a, it's basically a high fiber meal. We want you to have it for lunch three days in the week and then you have your control group where they just get the pills. And so I think by deliberately making sure that people are getting this added fiber, you can sort of compare whether that fiber is helpful.

It almost feels like a no brainer. The people who will have higher fiber food will do better, but you will measure their microbiomes and we'll see whether the strains are actually even higher for them too. So the next two years really is an interesting time for your company, but more importantly, I think just our scientific understanding of what's going on on the basis of a product your company has figured out how to make. So if nothing else, you guys have figured out how to make something from a manufacturing process that no one else was going to figure out. There's no university that could ever have

figured this out because they wouldn't have been able to put the resources in it. No one was going to spend $100 million to figure out how to make an obligate anorob when the maximum NIH grant is $480,000 a year. It's not going gonna happen. And now it enables a bunch of people to ask these questions about what's the impact on depression, what's the impact on ADHD, what's the impact on obesity, type two diabetes. It's super fascinating. And many ways it feels like the bridge

between the diet and the drug. Because we already have a really good sense of, like, it's not like it's rocket science, from an efficacy standpoint to get somebody to lose weight by changing their diet, the effectiveness is the problem. It's too hard to implement for most people. At the other end of the spectrum, we clearly know now how to do it with drugs.

And that's really changed. I think it really started in 2014 when Lyra Glutide came out, but clearly Semiglutide was the game changer. And that's three years ago, was when we saw the pivot from Semiglutide was the game changer, and that's three years ago, was when we saw the pivot from Semiglutide as just a diabetic drug, two-in-obey city drug. And it's exciting for me to just watch this. And frankly, even look at other questions that haven't been answered yet, like the dose response is more better. Do we know anything about how much is too much, and what's the maximal dose,

the minimum effective dose, and the median effective dose. There's all this other stuff that, I don't know, it'll be five years and we'll be sitting here and hopefully having a more interesting discussion. Absolutely. I think the dosing is interesting one, too, because the right question is, what is the colonization that's happening? Because it's not just about what you're delivering, it's about what's colonizing, and it's different from person to person. So I think if we could crack that nut on the dose

to colonization ratio and in what context that's better, you start to see improvements. But absolutely, it is the gap between nutrition and then these downstream small molecules. And the microbiome has been a black box. And now we've got some tools here. We're just at the beginnings of it. But I think for us, the big name of the game is, how do we get these products into as many people's hands and as many investigators' hands as we can to just create more and more data around what is your starting microbiome, what is your lifestyle,

how are these things tied together and what are the health outcomes that can really come from a different microbiome? Are there any other bacteria out there as specific strains or even species that you think are going to be worth investigating based on the literature today? And we've talked a lot about acrimansion,

all the reasons why you had so much data to stand on the shoulders of, to go down that rabbit hole. Are there others out there that you think offer promise and that maybe in five years there are many other strains out there that people are taking. So you might have in your fridge a bottle of acrimansia strain two, strain three, strain four.

Absolutely. I assume you're not gonna say what they are. I'm guessing that's some of the proprietary at this point. Absolutely. There are other, I think, of these sort of key strains that I'll just kind of allude to the link between our microbiome and our immune response. It's super interesting.

There are other strains. We'll stay tuned. All right, thanks, Colleen. Thank you so much for having me. Thank you for listening to this week's episode of The Drive. It's extremely important to me to provide all of this content without relying on paid ads. To do this, our work is made entirely possible by our members, and in return, we offer exclusive member-only content and benefits above and beyond what is available for free.

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