Neuroscience Expo: The Gut Bacteria and Brain Connection: Fix Your Gut and Fix Your Brain

We are honored to have Dr. Sharice Barvay, he is a professor of medicine in the Division of Medicine at the University of Louisville and gastroenterology, hepatitis and nutrition, he is also associate director of the Uofl Alcohol Research Center and director of the core bioanalytical Heptobiology, Toxicology and Cobra. he has recently established metagenomics integrated into metabolism. I should have studied this before Dr. Barbay Well excellent in microbiostructure and functional data that can be integrated with probiotics Clinical and interventional outcomes research An important component of Dr. Barbe's research program focuses on acquiring fundamental knowledge about the pathogenic changes in the access to the

brain

from the microbiotic gut and the application of this knowledge to reduce the burden of neurological diseases, in particular its research program employs basic clinical and translational approaches aimed at developing relevant preventive and therapeutic intervention strategies to curb cognitive decline and preserve memory function. destroyed that but please help me welcome Dr.

Sharice barbey Foreign thank you very much for the kind introduction welcome everyone. I would like to thank the Northern Neuroscience Institute, particularly Dr. Shields and Dr. Cooper, for inviting me to present our ongoing work. In today's talk, my intention. is to provide you with a conceptual framework that sets the tone for the gut

bacteria

in

your

system and their

connection

to the

brain

and how these

connection

s are significantly important for good health in the context of new regenerative diseases. Okay, sorry, neurodegenerative. diseases and how we can have intervention strategies to improve outcomes so as the title says fix

your

gut and fix your brain which is also a healthy gut it's a healthy brain so to start I'm going to give you information about what is the intestine.

The microbiome is in all of us, so what you see here is the elemental channel and as you go through the descending cone, here you have

bacteria

available in your stomach because there is high acidity. The pH is about one to two, but as you go down the elemental channel. The numbers start to increase from the duodenum jejunum helium and colon and when you get to the colon there are literally billions of bacteria. The trivia is that there are literally 10 to 15 times more bacterial cells in you compared to all your other sales as humans, so here's the trip.

Another very interesting piece of information is that if you look at the total number of genes, close to millions, more genes reside in your gut microbiome, which are housed in bacteria, and American humans have about 23,000 genes in total that make us tick, so You can see this incredible connection between the gut microbiome and your system and, as it says here, 99 of your DNA genes are in microbial cells and not in human cells, so you can clearly see the proportion and size of this organ liquid that is in us and at least 15 or 20 years ago we began to realize that this system clearly has to play a role in both physiology, I mean health and disease, and that is where the part is initial of the broth, so let's see what are the beneficial functions of the intestinal microbiota found in you, first of all, they have a very important role in regulating your metabolic function, for example, the fermentation of non-digestible substrates into products useful, so when you eat vegetables and salads, etc., period of indigestible fiber that is converted into some very useful powerful molecules or short chain fatty acids and I will show you some very interesting facts about how this helps you regulate prevention of diseases and maintaining good health biotransformation of bile acids our system produces a lot of bile that the liver produces and ends up in the intestine and the correct transformation again is very important for good health, since its production of vitamins some of the vitamins of the complex B are produced by our gut microbiome and then energy extraction, so there are many indigestible food elements that your healthy gut microbiome can extract energy and provide you with, most importantly there is a protective function and this is a resistance to colonization extremely important, meaning if you have a healthy health. diverse gut microbiome when you inadvertently have hostile bacteria, pathogenic bacteria, or disease-causing bacteria inside you, they're not allowed to take hold, so some people can travel anywhere, eat, no matter what, it doesn't happen nothing and some people can have diarrhea or, you know, have problems, so having a healthy and diverse microbiome is extremely important that offers you resistance to colonization, colonization of the bad bacteria, then there is antimicrobial secretion, your beneficial bacteria , good and friendly basically produce molecules that are not friendly to the bacteria, fungi or viruses that cause disease, so that is an important part and then it is very significantly the good bacteria or the right commensal flora continually educates your immune response when this crosstalk it breaks down and you'll see it further, that's where the inflammation, chronic inflammation, all those problems start and eventually there are structural. and histological aspects provided by your intestinal microbiome, which is the growth of epithelial cells and differentiation of the intestine.

There is an intestinal village and development and then a tight junction permeability. You must have heard the terminology of leaky gut, so if that should be prevented. because that's important, otherwise these systems get into the system and cause inflammation. Maintaining the permeability of the tight junction is extremely important and good bacteria make what are the components that life in general has as an effect and there are some of the important problems that The methods we review are listed here, starting from birth, the infant feeding methods are important, stress, exercise, metabolism, psychological stress, sleep, you will hear from Dr.

Shields after this talk about how sleep is very relevant when your circadian rhythms change, whether through , for example, jet. delaying international travel or whatever happens definitely affects your gut microbiome. Diet is clearly important if you eat a healthy diet with fiber and fruit sources. More processed foods. Alcohol has not been demonstrated here, but alcohol again. Past a certain point it can have a big impact on the gut microbiome Pharmaceuticals you've heard of polypharmacy with antibiotics so when you have things and you take antibiotics, those antibiotics not only kill the bacteria that are causing problems, but they also they start to affect the good bacteria, so polypharmacy is a very important component in geography, we have seen differences in people from Eastern Europe versus Asia versus within the United States from the south to the north, there are differences due to climate of diet, etc. and then obviously the stages or stages of the life cycle aging is known to change your microbiome as you progress from childhood. middle age and old age, so these are some of the components that have a huge impact on an ongoing basis on your gut microbiome, so let's talk about bacterial dysbiosis, so this is a term that you may have heard, which means it is change of the normal commensal flora in an altered form and that is called dysbiosis and this biases the first component of this bias is called reduced diversity, so we have more than 500 per species in our gut, but they are very diverse and it takes a diverse Flora as you start to get that diversity, which is a hallmark that your microbiome is changing and that's not good for you because now you're going to get diverted to where you can get an expansion of pathobions or bad bacteria can start to proliferate and the counterparts, you start to lose the beneficial bacteria, as I talked about the beneficial bacteria that convert indigestible fiber into short chain fatty acids, extract energy, produce vitamins, etc., so this bias is a collective term and only I want to point out that if you look at any disease state, from diabetes to rheumatoid arthritis, depression to alcohol literacy, there is dysbiosis, but it is contextual, meaning that the type of bias associated with diabetes is not similar to what is seen with Alzheimer's, so the whole area of ​​research is to identify very specific measures of these biases in a given clinical context so that we can solve it, that is the main aspect, as I just said, the intestinal device is seen in multiple different pathological states , you have hypothyroidism, there is muscle loss, rheumatoid sarcopenia. arthritis clearly IBD Crohn's diabetes cardiovascular diseases and clearly for today's talk I am going to focus on neurodegenerative diseases such as Parkinson's Alzheimer's multiple sclerosis depression anxiety pain and stress I am going to put special emphasis on Alzheimer's, but it starts from the same conceptual framework as in What we are working on is applicable to other forms of neurodegenerative diseases and this brings me now to the gut-brain axis, so the gut-brain axis is a bilateral communication network between the gastrointestinal tract and the central nervous system, so It's a two-way highway, okay?

The gut has an influence on the brain and the brain can also send signals to the gut, such as controlling motility, etc., and affect the gut microbiome. A vagal nerve or vagus nerve is a very important part of this bilateral connection, so in recent years there has been increasing clinical and preclinical evidence implicating the microbiome as a possible key susceptibility factor to neurological disorders that impure Alzheimer's disease, Parkinson's disease, autism spectrum disorder, multiple sclerosis and strokes, so again it is increasing, let's take into account the dysbiotic characteristics that are connected with these different disease states that are neurodegenerative , there are nuanced differences and part of the job is understanding what is relevant to Parkinson's, they may share a lot of commonalities, in fact they do, but there are some very specific and very nuanced changes. that then connects to Parkinson's versus Alzheimer's and that's what we're trying to figure out and we have some answers right now, so this leads to the general hypothesis of our work that, like I said, we do it with different ways of lifestyle problems, stress. lack of sleep, as you'll hear in the next talk, antibiotics, you know, the mode of administration, etc., can affect your microbiome adversely, while probiotics, exercise can also affect your microbiome, but in the good sense, once you have this disturbed microbiome you sit down.

As I talked about the loss of diversity, you have a kind of spiral, there could be an increase in pathogens or disease-causing bacteria and then the loss of beneficial bacteria, once this sets in, it affects the intestinal wall and there is an increase in the intestinal permeability that begins to be lost. the barrier function and then the microbial products or antigens as we call them start to leak into your system so what you are seeing here is the intentional lining starting to open up and the microbial products start to enter your system once that enter. drives all kinds of pathogenic events, the main event is inflammation and this is something that is of great value, for lack of a better word, I'm going to say it is a smoldering fire that continues, you know, released by the intestine and this . is where your end organs will be stressed in our context, we are trying to understand and appreciate what neuroinflammation and neurodegeneration means and, very interestingly, the same proteins that keep your gut barrier intact are the same proteins in your blood brain barrier, so which when you see a gap in your intestinal permeability, it also changes the permeability of the blood brain barrier, so now your system, whatever it's oozing out, is starting to affect the brain and I'll show you some evidence of all of that and again once you start triggering this process in the brain, this is the General Motors operation that comes into play and is involved in Alzheimer's, Parkinson's, Huntington's disease and amyotrophic lateral sclerosis, so all of these different forms of neurodegenerative diseases, we have no evidence to show that there is a significant change in the gut microbiome there is a significant change in the system of molecules that are affected by the change in the gut microbiome, so it's like a domino effect that starts As I told you, I'm going to focus a little bit more on Alzheimer's for today's talk and if we now take the gut-brain connection specifically in the context of Alzheimer's.

What is the background? What do we know? Recent clinical research has shownthat, compared to sex- and age-matched control individuals, Alzheimer's patients have altered gut microbiome dysbiosis showing distinct pectoral composition and significantly decreased diversity, so these are established facts, that is the important evidence . Alterations in gut microbial composition in Alzheimer's patients include an increased abundance of pro-inflammatory bacterial taxa and a concomitant decrease in anti-inflammatory taxa. So these things are now being detailed and the reason for detailing this is in the hope that we can fix it, so overall the current research strongly supports the postulate that the gut microbiome is involved in the initiation and exacerbation of pathology of Alzheimer's disease and clearly more and more work is being done so I will stop here and now we want to understand when we say that this gut despises and Alzheimer's is a mere association or the change of the microbiome is playing a causal role now if it is playing a causal role, it stands to reason that if we take care of it, fix it, it should have a beneficial impact and it should start to reduce the severity of the pathology, okay, either prevent it or cure it, like in a mature way, you are on a spectrum, so I'm going to present a clinical trial and there are quite a few that are emerging right now where they looked at the effect of probiotic supplementation on cognitive function and metabolic status in Alzheimer's disease, so it was a very useful trial and I'm going to present Here's some of the hard evidence, so first of all, this study was a 12-week randomized, double-blind, controlled clinical trial, which means that when the patients came in, they were randomized, half of the patients Patients received a placebo and the other half received a probiotic supplement and then they were compared because for any true effect, their intervention would have to be greater. than the placebo because otherwise it's just a placebo effect, so let's revise this so that the patients were randomly divided into two groups n equal to 30 in each group, treating them with milk, which was the control group, or again with a probiotic mix that was in the milk. which changed the composition of the milk and included the probiotic bacteria, so the probiotic supplemented group took 200 ml per day of probiotic milk containing lactobacillus acidophilus lactobacillus KSI bifidobacterium bifidum and lactobacillus fermentum now these are just names but it is identified that These bacteria have a beneficial value or all the details of their genetic expression, what they produce is all detailed, that is why they are included here and the dose is quite large, so each type of bacteria is approximately 2.10 to the power of 9 , it's 2 billion of each type, we're including the milk, so when the patients drank plain milk or milk with these bacteria and this duration was 12 weeks, let's see what the end point is here, so the end point is very encouraging, after 12 weeks of intervention compared to the control group. in the patients treated with probiotics showed a significant improvement in the primary outcome which was cognitive function and this cognitive function was assessed by the mini mental state examination score and this is a very important finding that the cognitive scores improved after 12 weeks in these patients. the other very important secondary outcome because all the secondary outcomes are also important, which were changes in the metabolic function of these individuals, particularly Melania aldehyde in plasma, is an indicator of oxidative stress in their body, so the series of highly sensitive active proteins in serum.

It is a marker of inflammation. It is an acute phase reactant that descends from Homa ir and Homa B, which reflects the ability to control glucose, so in the conditions of prediabetes or diabetes, those indicators improve and finally serum triglycerides and the quantitative control index of insulin sensitivity. they also improve so in this trial the important finding was that in 12 weeks the cognitive scores improved significantly and also had a very beneficial impact on the metabolic outcomes now if I have to I won't say criticism but now these are small studies that We need big studies here as you know this gives us proof of principle that hopefully gut problems have this and this also proves the point that there is a connection between the gut and the brain so the bottom line is a bit.

I clearly stated that probiotic supplementation demonstrates some encouraging trends that Warren will study further to evaluate whether probiotics have a clinically significant impact on cognitive symptoms in Alzheimer's patients, so this is one of the earlier trials and as I will discuss some of our ongoing plans, we are also planning some large evidence-based clinical trials, hopefully very soon in the near future. From the clinical studies, I'm going to show you some very interesting data that we've been getting from the preclinical model. It's important to do preclinical work because it allows us to do a lot more things and then translate that to patients, so preclinical work gives us not only proof of principle validation of some of the concepts that we can examine in a very robust way and then Know what the pressure points are and then get there.

Going back to patients, in this case we are looking at the gut-brain connection by using it in an animal model of Alzheimer's and this animal model is extremely interesting and exciting. First, what was the intention of the preclinical work: to investigate the causal relationship. role of intestinal dysbiosis in the development of Alzheimer's disease progression these are the animals that suffer from mechanical Alzheimer's disease because they are made to do so and then we wanted to determine the impact of evidence-based management of intestinal microbial biases in the prevention and treatment of Alzheimer's disease is found in these animal models and as you will see, we are provided with a treasure trove of information that can hopefully be translated to people, so this is called triple trans single, forget the words. uh three xdg mouse model and it's a triple transgenic Alzheimer's mouse that expresses the mutant of these three genes that are listed here that are human transgenes and in the brain and then basically allows these mice to recapitulate specific aspects of neuropathological progression. of Alzheimer's disease, including age-dependent cognitive decline occurs.

There is accumulation of neuronal plaques and neurofibrillary. Age-dependent tangles and inflammation. He is fine, most importantly, because of his phenotype. These are a valuable tool to investigate the molecular mechanisms underlying the different stages of Alzheimer's disease. It allows us to observe. The genes that are involved play out what is happening and dissect the whole system very clearly. This is the time path from one month to 18 months to 18 months. All of these animals have full-blown Alzheimer's. They start in part. Cognitive decline begins in the youngest. At three months, the amulet plates, uh glucis, begin to form neuronal tangles and then you will see that there is a decrease in behavior in these animals, so it is a very useful model to investigate your conceptual intervention strategies.

The first thing we did and analyzed it. gut microbiome, as you are seeing here, which depends on age, from two months to 18 months, just the color coding, forget about the names of the bacteria, the color coding tells you that the microbiome is changing or aging and clearly At 18 months there is a significant difference. varicomicrobiota gain uh, there is a reduction in the use of firmic and there is a bacterial diet. He tells me he would go into detail about the microbiome where animals started and that also happens to people as you get older, so your microbiome keeps changing. and this is the first indicator that, and again I just showed you the trajectory, they are also developing Alzheimer's pathogenesis as the microbiome is changing just a small detail.

Bear with me, what we see is the firmicutes edge at the beginning. here the firmicutes edge diminishes. It harbors a group of beneficial bacteria, particularly butyrate-producing bacteria. Now I'll show you that and the bacteria are actually increasing, so when you take a ratio between these good phyla and the bad ones you see a drop in the ratio. and this is a very significant telltale sign that diversity is changing and the gut microbiome is going through a significant attraction or change, so then we focus. I won't go into details because of the use of firmic, we had other evidence to show that the bacteria in us that produce butane or less fatty acids are clearly very relevant and if you look at that here you can see that they have a pretty large complement of butler producing bacteria which starts to decrease with age and again I have this timeline in front of you of how they develop through the pathogenesis of Alzheimer's the color coding shows you the different types of butyrate producing bacteria.

There are at least 265 known species that produce butyrate for you and again, starting at two months, there is a very significant drop or change in the butane-producing bacteria. So I showed you a drop in diversity and it shows also when you look at the reason why the diversity is decreasing, they are also losing beneficial bacteria very quickly like in a cartoon here on the left side here this is a healthy section of the intestine , here is a healthy looking intestinal epithelium, there is a significant amount of healthy mucous layer that does not allow bacteria to come into contact with the intestinal epithelial cells, you have the right type of bacteria that will convert it into For example, indigestible fiber for rate and all of these things go into your system and this is called the lamina propria, it's a loose connective tissue that surrounds the elemental canal and this is where most of your immune cells keep coming to slurp up what's in the gut. it means you know the direction, you are what you eat in a sense, so your immune response now this is where they come, or they are patriots, they just become terrorists depending on what they feel from the gut, if you look to the right side, This is a general. issue that is seen in multiple pathological states, clearly there is loss of mucus, the bacteria are now not friendly, they are more pathogenic, not the beneficial bacteria have decreased, there is what we call paracyllular permeability that is opening the intestinal tract and the system and the microbuilding products and the antigens come in and once that happens, it starts sending out inflammatory triggers and that's a bad thing to happen so quickly to connect that aspect to the brain.

I talked about short and fatty acids. significant work to show that the good bacteria that produce short, greasy pants have this particular butyrate. I would go and bore you with the details, clearly there is molecular evidence through chemical real scientific evidence that shows how it can influence what happens in the brain. so the thought was once you know this, how do we change this equation? And in a nutshell, we embarked on a molecule called tributarin, which is a butyric prodrug and this is a triglyceride that contains three butyrate moieties esterified with glycerol, it is a natural molecule and it is present. in a variety of foods, particularly dairy products like butter, and is metabolized in the intestinal lumen by pancreatic liposes and the resident intestinal microflora, so we decided to give tributin and ask: can we address the problems that we are seeing?

And is it relevant to the progression of Alzheimer's disease? So I showed them this trajectory and we started giving tributarin supplements at six months of age to these animals and obviously there was a group of animals that did not receive tributarin oil supplements and let's take a quick look. Look at what happens, first of all, if you compare the 18-month-old animal that has full-blown Alzheimer's and then the animals that started receiving Tributan at the age of six months, you will see a very significant change in the composition of the gut microbiome. See with the color-coded bar chart here, these are all measures of diversity.

I would go into details, but clearly the charm index, the Shannon index, the Simpson index, in each aspect we began to see a change, the phylum firmicus that was downward, as I showed. you went up through the backyard it's a file cabinet that went up went down and the ratio has now increased very significantly so it definitely told us that the tax administration is changing the equation in the gut at the level of microbial diversity and what we are showing here Also The question is to change the equation for butrada producing bacteria and the answer is yes, this year, 18 months, where the butrada producing bacteriaButler have been largely lost and look what happened to the animals that received drybutrin for six months and were able to preserve them. beneficial bacteria and these are the total butane producing family, so this is where we quantify and see a very significant change.

Now it was time to go and see the brains of these animals or the brains of these animals to see what happens after this. This was very exciting, so it's two months for healthy animals, 10 to 12 months times 18 months. Okay, if you see these green dots that talk about neuronal inflammation, then the microglia express iba1, so there is significant neuronal inflammation, but now compared to the next one. slide here, which is 18 months in which they were administered tributarin, we saw that neuronal inflammation was significantly prevented, it did not happen in these animals now neuroinflammation is a very important factor in the pathogenesis of Alzheimer's disease, it is a diode for other forms of neurodegenerative diseases as well and what you see here is a quantification of this data which is a very significant increase from two months to 18 months, but now compared to the same animals at 18 months but receiving tributin starting at six months of age, a significant reduction in neuroinflammation, these were the most interesting data in this and there are several other data points that I'm not going to show, but you must have heard of uh to beta, but the most important part is the protein Hyperphospholic tau that forms neurofibility.

Tangles that are neurotoxic, so when we gave, it's two months there. There is practically no action. Here is your 12 month old animal. Your 18 month old animal is particularly focused on these yellow hyperphosphorus Tau and when we looked at the animals that had received tributin, there was almost complete prevention of the neurofibular angles that are neurotoxic and that are happening, so the result. was that this supplementation prevented the formation of neurotoxic tangles of hypo or hyperphosphate and the next slide is to show you the quantification because what I was showing you is a representative slide, look at the increase at 18 months, these are the animals that have all their power.

Alzheimer's and look at the counterparts we see in private, the Tau hyperfossil accumulation was almost completely blocked, what does this mean? Oh, one more data point that is very important, so if you look at the beginning, this supplementation prevents, it's a little technical, the deacetylation of histones that are required for active gene expression, so these animals here see a lot greener, okay, that's starting to fade and that's because they're losing histone acetylation, which that also tells me that histone scaling is controlling gene expression, so there are several gene functions that are lost with age. and, as you know, that's where they also get the neurodegenerative disease, but look at the same animals. who receive tributin, this loss was remarkably protected, which tells us that it is also protecting the genes from being diluted or inhibited or aged, so what does this mean on the cognitive behavioral end?

So first we look at the short term. The memory assessment in these animals is called the novel object recognition test. You place the device called habituation and then you place them in the second cage where there are two objects, and once they get used to those two objects, you remove one object and place a new one, the mice, instinctively, are normal, we will go to the new object that is called a novel object recognition test, but animals that have begun to suffer cognitive decline don't care much, they just know what it is and those patterns are tracked by a camera. are here, these movements are tracked bicameral Quantified and the quantification is shown here, so this is the discrimination index, which means it tells me how much they are discriminating between the two objects and again there is a young animal of 10 to 12 months and buy In green, you see a strong decrease in this discrimination index and what you see is that the animals that received tribute from six months onwards were able to preserve this function very well and we saw that this was repeated in their preference index and also in the recognition index.

Clearly, the animals that received tributin were able to prevent or we were able to prevent the age-related decline in short-term memory, so this was a really exciting finding. Then we do what's called a Y-maze test, which is an assessment of spatial memory and behavioral alternation. So it's a maze, so initially the animals are allowed to go and they will travel through all these mazes that you have and for the novel Army closes this novel arm and then they are allowed to move only in these two but one time. You open this, preferably they go to the novel that is open because it was not available to them, as you can see here, that is a good spatial memory that mice remember, oh, that was not accessible now it goes there preferentially, while the animals that They have Alzheimer's, uh, they don't because it's the same thing, they can't distinguish between opening a new arm and the same thing, as you can see, there's a big drop at 18 months and clearly this drop in spatial memory or alternation of behavior was prevented very significantly, so they are very interesting findings and again let me remind you that what is connected to this is the availability but availability rate of higher levels of beneficial bacteria and a more diverse bacterial flora, so who started testing causal connections because we're not giving this to the brain they're giving tributan as an oral supplement, so it's affecting the microbiome and then it's affecting the gut brain axis, another very important age-preserving part.

Associated decrease in false neuromuscular function. Fragility and fracture are a very important component of aging. There is documented evidence of loss of neuromuscular function in Alzheimer's associated, so this is a test where you place the animal on the grate, grab the grate and pull on it and recognize the grip strength and how firm the grip is. if you look again. In the grip strength here in the quantification there is a significant drop in grip strength, but look how we were able to preserve the grip strength or the neuromuscular function in these animals, so that is the grip strength, which is the strength and This is a Force that animals grab with, so in both accounts we saw a very significant improvement in the animals or preservation of neuromuscular function that received tributin starting at six months and lastly, I'll show you which prevents age-related decline in motor coordination. and the activity is a rotor rod, the animals stay here and it starts to move slowly and we try to calculate how long the animal can hold on or fall, as you can see the animals with Alzheimer's fall very quickly, wrote The Lord Falls. but look again at the comparison with the tributin supplemented group, so this was very significant, not only the memory part but also the neuromuscular components were protected very significantly and there is a lot of literature showing that the good bacteria and the Inflammation will affect muscle strength and all sorts of things, so in conclusion, intestinal microwave, this bias distinguished by a decrease in butogenic potential, is a key pathogenic characteristic associated with the development of brain pathology relevant to pathogenesis of Alzheimer's disease and the progression of the disease, so this is a very significant finding that we have made.

What are the specific events that need to be addressed and, more importantly, does the data provide a clinical rationale for targeting the gut? This bias mediates the loss of butogenic potential for preventive and therapeutic strategies to preserve or even improve neurological function in Alzheimer's disease and, in the end, I want to Basically, we presented our collaboration at the Northern Neuroscience Institute and this is our initiative of investigation that is about to begin. The intention is to gain fundamental knowledge about pathogenic changes along the gut and brain microbiota axis and apply that knowledge to reduce the burden of neurological diseases. disease and conduct longitudinal studies and randomized control clinical trials in well-defined patient cohorts targeting the gut microbiome with the aim of developing new preventive and therapeutic strategies.

I am grateful for my collaboration with Dr. Shields and Dr. Cooper. We're really looking forward to taking our entire knowledge base and giving it back to the people. Lastly, these are my collaborators at the University of Louisville. My other collaborators are at the University of Florida in Miami, with a significant Alzheimer's population that you are working with. Baylor College of. Medicine we would work on the analysis of the microbiome at Vanderbilt University and now, as I just mentioned the Northern Neuroscience Institute and finally I leave you with this, Hippocrates is considered the father of modern medicine, he said that all diseases begin in the intestine, poor digestion, digestion is the root. of all evil and death resides in the bowels thank you very much for your time foreign